27th Conference of the World Association for the Advancement of Veterinary Parasitology JULY 7 – 11, 2019 | MADISON, WI, USA Dedicated to the legacy of Professor Arlie C. Todd

Sifting and Winnowing the Evidence in Veterinary Parasitology

@WAAVP2019 @WAAVP_2019 Abstract Book

Joint meeting with the 64th American Association of Veterinary Parasitologists Annual Meeting & the 63rd Annual Livestock Insect Workers Conference

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 1 63rd Annualwww.WAAVP2019.com Livestock Insect Workers Conference #WAAVP2019 Table of Contents

Keynote Presentation 84-89 OA22 Molecular Tools II 89-92 OA23 Leishmania 4 Keynote Presentation Demystifying 92-97 OA24 Nematode Molecular Tools, One Health: Sifting and Winnowing Resistance II the Role of Veterinary Parasitology 97-101 OA25 IAFWP Symposium 101-104 OA26 Canine Helminths II 104-108 OA27 Epidemiology Plenary Lectures 108-111 OA28 Alternative Treatments for Parasites in Ruminants I 6-7 PL1.0 Evolving Approaches to Drug 111-113 OA29 Unusual Protozoa Discovery 114-116 OA30 IAFWP Symposium 8-9 PL2.0 Genes and Genomics in 116-118 OA31 Anthelmintic Resistance in Parasite Control Ruminants 10-11 PL3.0 Leishmaniasis, Leishvet and 119-122 OA32 Avian Parasites One Health 122-125 OA33 Equine Cyathostomes I 12-13 PL4.0 Veterinary Entomology: 125-128 OA34 Flies and Fly Control in Outbreak and Advancements Ruminants 128-131 OA35 Ruminant Trematodes I Oral Sessions 131-135 OA36 Treatment and Control of GI Nematodes in Ruminants 15-18 OA01 Canine Heartworm I 136-139 OA37 Poultry Coccidia, Aquatic 18-21 OA02 Diagnosis and Decision Infections Support for GI Nematodes in 139-144 OA38 Equine Cyathostomes II Ruminants I 144-148 OA39 Insecticide and Acaricide 21-24 OA03 North American Ticks Resistance in Ruminants 25-28 OA04 Coccidia 149-152 OA40 Zoonoses 28-30 OA05 Worldwide Vector-Borne 153-155 OA41 Biology and Pathology of GI Infections in Companion Animals Nematodes in Ruminants 30-35 OA06 Canine Heartworm II 155-158 OA42 Diagnostic Techniques 35-38 OA07 Host Responses Against 159-161 OA43 Equine Parasites Helminths in Ruminants 161-164 OA44 Canine Arthropods 39-42 OA08 Tick Disease Transmission 164-167 OA45 Ruminant Trematodes II 43-46 OA09 Wildlife Parasites 168-171 OA46 Gastrointestinal Protozoa in 46-49 OA10 New Tools and Big Data for Ruminants Evaluating Intestinal Parasite 171-175 OA47 Wildlife Helminths Infections in Companion Animals 175-179 OA48 Equine Ascarids 50-52 OA11 Canine Protozoa 179-183 OA49 Ticks on Cattle 53-56 OA12 Diagnosis and Decision 183-187 OA50 Alternative treatments for Support for GI Nematodes in Parasites in Ruminants II Ruminants II 56-59 OA13 Flea and Tick Treatment Poster Sessions 60-62 OA14 Protozoan Parasites 62-65 OA15 Education 189-234 PS01 Poster Session 1 65-68 OA16 Canine Helminths 234-280 PS02 Poster Session 2 68-71 OA17 Molecular Tools I 280-326 PS03 Poster Session 3 71-74 OA18 Leishmania 74-78 OA19 Nematode Molecular Tools, Resistance I 78-80 OA20 IAFWP Symposium 80-84 OA21 Cat Parasitisms

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 2 63rd Annual Livestock Insect Workers Conference KEYNOTE PRESENTATION

With an overall theme of Sifting and Winnowing the Evidence in Veterinary Parasitology, the Conference reminds us that we cannot for a moment believe that knowledge has reached its final goal. We honor the tremendous contributions to veterinary parasitology by University of Wisconsin Emeritus Professor Arlie C. Todd, mentor of 59 graduate students, 44 of which earned a Ph.D.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 3 63rd Annual Livestock Insect Workers Conference KEYNOTE PRESENTATION

Keynote Presentation

Keynote Address and Dedication Parasitology”. This medical doctor and Demystifying One Health: Sifting and scientist will explore how parasites move Winnowing the Role of Veterinary between animals and people and current Parasitology status of clinical information sharing between human and animal clinicians. He will compare July 7, 2019, 13:30 - 15:30 clinical diagnostics for parasitic diseases in Plenary Hall, Madison Ballroom (ABCD), Level 4 humans, animals, and barriers to information sharing using Giardia as example. This discussion will describe the microbiome as a Demystifying and Demonstrating the Value crossing point between humans, animals and of a One Health Approach to Parasitological environments. Challenges c. Patricia A. Conrad: “Babesia and Professor Rosina C. Krecek1, Professor Toxoplasma Discoveries at the Human-Animal- Patricia A. Conrad2, Professor Peter M. Environment Interface”. As a veterinary Rabinowitz3 protozoologist and scientist she will describe 1Global One Health, College of Veterinary how a One Health approach transformed her Medicine and Biomedical Sciences, Texas A&M research and resulted in surprising discoveries University and University of Johannesburg, of new parasite species and the impact of South Africa, College Station , United States, environmental factors on protozoal pathogen 2Department of Pathology, Microbiology & transmission. Lessons learned in challenging Immunology, School of Veterinary Medicine, the dogma in protozoology, working with University of California, Davis, United productive teams, and making parasitological States, 3Departments of Environmental and research fun will be shared. Occupational Health Sciences, Global Health, Family Medicine, University of Washington This session will illustrate an interdisciplinary Center for One Health Research, Seattle, One Health approach in veterinary United States parasitology to encourage research, education, and a scientific basis for local and A panel of three One Health advocates will global health policy decisions. Further, the illustrate with zoonotic parasite cases, key need to improve quality of evidence-based principles, skills and outcomes what they have metrics, development of useful tools for learned from applying a One Health approach tracking One Health outcomes and better to medical and veterinary clinical practice. ways to assess impact will be explained. a. Rosina C. Krecek: Title: “Pork Tapeworm: A One Health Model”. This veterinary scientist educator and researcher will illustrate how Taenia solium, a neglected tropical helminth parasite, infects pigs and 50 million humans across developing countries, and is preventable with tools available today. This disease limits pig production, causes human epilepsy and threatens food security necessitating a new One Health approach that incorporates vaccination, antiparasitic therapies and educational materials. b. Peter M. Rabinowitz: Title: “A Clinical Perspective on One Health Challenges for

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 4 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 5 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

Plenary Lectures

Plenary Lecture 1.0 Evolving PL01.02 Target-Based Drug Development for Approaches to Drug Discovery Cryptosporidiosis and Other Apicomplexa Diseases July 7, 2019, 16:00 – 18:00 Plenary Hall, Madison Ballroom (ABCD), Level 4 W.C. Van Voorhis1, M.A. Hulverson1, R. Choi1, W. Huang3, S. L. M. Arnold1, D. A. Schaefer2, D. P. Betzer2, R. S. R. Vidadala4, G. R. Whitman1, L. PL01.01 Data-Driven Revolutions in K. Barrett1, D. J. Maly4.3, M. W. Riggs2, E. Fan3, Antiparasitic Drug Discovery and K. K. Ojo1. 1Department of Medicine, Division of Allergy Prof. Timothy Geary1 and Infectious Disease, Center for Emerging 1Institute of Parasitology, McGill University, and Reemerging Infectious Disease (CERID), Ste-Anne-de-Bellevue, Canada, 2Queen’s University of Washington, United States University of Belfast, Belfast, Northern Ireland, 2School of Animal and Comparative UK, 3CryBioMed, Philadelphia, United States Biomedical Sciences, College of Agriculture and Life Sciences, University of Arizona, Parasite infections remain major and global United States impediments to human and animal health. 3Department of Biochemistry, University of Control measures involving environmental Washington, United States interventions have led to significant 4Department of Chemistry, University of improvements in the prevention of some Washington, United States of these infections, but are necessarily supplemented with chemotherapeutic Bumped kinase inhibitors (BKIs) have shown options. For many parasites, available drugs promise in animal models of cryptosporidiosis, are less than optimal and drug resistance toxoplasmosis, neosporiasis, and poses real and potential threats. New sarcocystosis. They were designed to target developments in bioinformatics and functional calcium-dependent protein kinase 1 (CDPK1), genomics – addressed to various extents in an enzyme for these organisms that has ‘big data’ science – are poised to revolutionize descended from the plant kingdom and our understanding of parasite biology and appears to control cell entry and replication of our ability to manipulate the expression these obligate intracellular parasites. Using of specific parasite genes in ways that the knowledge from the co-structures of facilitate the discovery of new antiparasitic CDPK1 and BKIs, improvements have been drugs. These drugs are likely to improve the made to the safety and efficacy of BKIs, and treatment of parasitic infections of both they are advancing toward human and animal human and veterinary importance and will use for treatment of cryptosporidiosis. As the include agents that influence the interface to understanding of BKI pharmacodynamics for favor the host. Improvements in diagnosis cryptosporidiosis therapy has increased, it will enable us to move from very broad- has become clear that better compounds for spectrum agents to more targeted treatments efficacy do not necessarily require substantial in the future, potentially changing the rate of systemic exposure. We now have BKIs with resistance selection. Research that enables reduced systemic exposure, acceptable better insights into fundamental aspects of safety parameters, and efficacy in both the parasitism can be anticipated to pay off in the mouse, newborn calf, and piglet models of discovery and development of better ways cryptosporidiosis. These BKIs are promising to control these pathogens, and we must pre-clinical lead for cryptosporidiosis therapy take full advantage of new science and new in animals and humans. Since toxoplasmosis, technologies to make this a reality. neosporosis, and sarcocystosis require more systemic distribution, we have alternative BKIs that display superior systemic distribution and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 6 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

safety properties, and have efficacy in mouse classes repetitively revolutionized our models of these diseases. These are coming understanding of state-of-the-art antiparasitic under later stage investigations in farm animal chemoprophylaxis/chemotherapy and thereby models. also re-defined the expectations towards future generations of effective, safe and Supported by US NIH/NIAID R01AI089441, convenient antiparasitic products. R01AI111341 and R01HD080670 and USDA In simplified terms, antiparasitic drug 2014-06183. discovery was historically performed by testing compounds, sometimes compound libraries, against a pathogen of interest in PL01.03 Excretory/Secretory Products vitro, followed by an in vivo validation in of Nematode Parasites: Significance and infected animals. Once proof of concept Therapeutic Implications against a particular pathogen was demonstrated, both parasite and host species Dr. Lucienne Tritten1 were expanded to evaluate the spectrum 1University of Zurich, Zurich, Switzerland of efficacy and safety. This approach resulted e.g. in the discovery of the endecto- Parasitic nematodes are highly successful parasiticide ivermectin, one of the top-selling pathogens, inflicting diseases on humans, veterinary medicines in the world. animals and plants. Despite great differences in their life cycles, preferential hosts and However, the evaluation of approaches to transmission modes, these parasites share a tackle parasites became and still becomes common capacity to manipulate their host’s more diverse and creative. Novel approaches immune system. This is known to be achieved include high-throughput phenotypic or target- through the release of excretory/secretory based screenings of large compound libraries, proteins, the most well-characterized identification of cellular and molecular component of nematode secretomes, pathways and potential targets via –omics which are comprised of functionally diverse approaches, re-evaluation of traditional molecules. Recently, among others, non- medicines, repurposing of old drugs alone coding RNA has been proposed as potential or in combination, vaccination strategies, new player in host manipulation. but also consider breeding, microRNA or This lecture will summarize the current gene therapy as potential antiparasiticide status of research on excretory/secretory innovations of the future. products of nematode parasites and their expected biological significance, with a focus However, once an efficacious drug candidate on proteins and microRNA. Furthermore, has been identified, complexity is added: how molecular mediators of host-parasite the future drug needs to be safe for target interactions may be translated into new animals and humans, needs to be suitable targets for therapeutic interventions will be for formulation, and should have favorable discussed. pharmacokinetic properties and reasonable production costs, to mention just a few aspects.

PL01.04 Next Generation Parasiticides: A The purpose of this presentation is to Mission We Can Accomplish Together share not only the hurdles and challenges but also the enormous potential and great Dr. Daniel Kulke1 opportunities towards broadening the 1Bayer Animal Health, Germany antiparasitic treatment options of the future for both veterinary and human medicine by Following more than 100 years of antiparasitic all the great achievements contributed by drug discovery, Bayer currently celebrates academia and pharmaceutical industry as its centenary of animal health dedicated well as numerous governmental and non- antiparasitic products with the launch of governmental organizations and initiatives Neguvon for the treatment of scabies in 1919. as part of our worldwide community for the Since then a series of chemical drug advancement of veterinary parasitology.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 7 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

Plenary Lecture 2.0 Genes and PL02.02 Extracellular vesicles: A Conserved Genomics in Parasite Control Nematode Strategy for Host Manipulation?

July 8, 2019, 8:30 - 10:30 Dr. Wang Yuan1, Ms Hannah Loghry1, Dr. 2 1 Plenary Hall, Madison Ballroom (ABCD), Level 4 Mostafa Zamanian , Dr. Yoko Ambrosini , Dr. Karin Allenspach1, Dr. Jonathan Mochel1, Dr. Michael Kimber1 PL02.01 Enlisting Spatiotemporal 1Iowa State University, Ames, United States, Transcriptomics to Identify Nematode 2University of Wisconsin-Madison, Madison, Receptor Drug Targets United States

Dr. Mostafa Zamanian1 Parasitic nematodes have evolved diverse 1University of Wisconsin-Madison, Madison, evasion mechanisms to manipulate host United States biology and create conditions that promote parasitism. Although the importance of Filarial nematode parasites continue to pose parasite excretory/secretory products in host a major challenge to animal and human modulation is well-recognized, we currently health, and new treatment options are needed have a less developed understanding of the for both heartworm and human filariasis. specific molecules parasitic nematodes use Transcriptomic tools are essential for the to manipulate the host, how those molecules discovery of potential therapeutic targets are trafficked to host cells, and how they that facilitate the development and survival of manipulate host cells at the genetic and parasitic nematodes within their verterbrate cellular level. We, and others, have identified hosts. Transcriptome studies in filarial that parasitic nematodes actively secrete nematode parasites have thus proved useful modulatory proteins and small, regulatory in detecting bulk changes in gene expression RNAs (miRNAs), and deliver them to host throughout the parasite life cycle and as tissues via extracellular vesicles (EVs). These a function of chemical or environmental EVs are a mechanism by which discrete perturbation. However, localization of gene populations of parasite-derived effector transcripts to specific cells or tissues has molecules can be protected from the host yet to be realized at an appreciable scale. environment and trafficked to host cells where Established in situ methods of gene transcript they exert their effects. Here we describe how localization are low in throughput and filarial nematodes (Brugia) prodigiously release complicated by the lack of fully defined cell EVs throughout their life cycle that contain and neuronal architecture in filarial parasites. a stage- and sex-specific cargo enriched in Further, single-cell methods to associate gene proteins with known and putative modulatory expression with individual cells can often functions. RNA-seq analysis reveals an EV be prohibitive in terms of costs and tissue miRNA complement that includes parasite requirements. In light of these limitations, we miRNAs identical to host immunomodulatory show how spatial RNA sequencing and single- miRNAs. The EVs are internalized by vector cell approaches can be optimized and enlisted and definitive host cells and we will discuss to generate high-resolution transcriptional potential host modulation at the transcriptional maps in filarial nematode parasites. We level. Gastrointestinal nematodes (Ascaris) discuss how these methods can be used also release EVs that, in comparison to filarial to prioritize putative receptor drug targets nematodes, contain a more complex cargo associated with parasite sensory behaviors of proteins that also includes putative host and secretory function. Together, these modulators. The miRNA cargo of Ascaris EVs data help provide the first steps towards a shares similarities to that of Brugia, suggesting spatially resolved cell and transcriptome map conserved mechanisms of host manipulation. in a filarial nematode parasite and present We will describe how Ascaris EVs are a roadmap that may be extensible to other internalized by host mucosal epithelium to clinically relevant parasitic nematodes. drive the earliest determinants of the mutually beneficial Type 2 response using a canine enteroid model. We conclude that investigating

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 8 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

the structure and function of nematode EVs PL02.04 Deep Amplicon Sequencing will generate new mechanistic understanding as a Powerful New Tool to Screen for of how parasites modulate the host at the Sequence Polymorphisms Associated genetic and cellular level. With Anthelmintic Resistance in Parasitic Nematode Populations

PL02.03 A Multimodal Approach to Dr. Russell Avramenko1, Dr. Elizabeth Understanding Macrocyclic Lactone Redman1, Lynsey Melville2, Dr. Janneke Wit1, Resistance in Dirofilaria Immitis Camila Queiroz1, Dr. Dave Bartley2, Dr. John Gilleard1 Dr. Cassan Pulaski1 1University of Calgary, Calgary, Canada, 1UGA, United States 2Moredun Research Institute, Penicuik, United Kingdom

Parasitic gastrointestinal nematodes contribute to significant human morbidity and cause billions of dollars per year in lost agricultural production. Control is largely dependent on the use of anthelmintic drugs, which in the case of livestock parasites, are severely compromised by the widespread development of drug resistance. Consequently, there is an urgent need for scalable and accurate diagnostic tools to detect the emergence of anthelmintic resistance particularly in its early stages. These diagnostics are often hindered, by the need to discriminate between multiple co-infecting parasite species, each with varying levels of anthelmintic resistance. Detecting and measuring the frequency of resistance-associated mutations in mixed parasite populations has the potential to provide sensitive and quantitative assessment of resistance emergence from an early stage. We describe the development and validation of deep amplicon next-generation sequencing as a powerful new approach to detect and quantify the frequency of single nucleotide polymorphisms (SNPs) associated with benzimidazole resistance. We have used parasite communities in sheep, which provide an excellent system in which to undertake the proof-of-concept of this approach, as there are multiple co-infecting trichostrongylid nematode species, each with varying levels of benzimidazole resistance. We demonstrate that this approach provides an accurate measure of resistance allele frequencies, and can reliably detect resistance alleles down to a frequency of 0.1%, making it particularly valuable for screening mutations at the early stages of resistance. We cover several examples to illustrate the use of this technique to screen for benzimidazole resistance, both

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 9 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

when resistance is already well established, need to be investigated. Moreover, limited but also as resistance is emerging in parasite information is available regarding the impact populations. This approach provides a powerful of canine vaccination in reducing human new tool to screen for the emergence of leishmaniosis both in Europe and in Brazil. anthelmintic resistance mutations in parasitic Another important use of vaccination is as nematode populations of both animals and immunotherapy to reduce the progression humans. of disease in infected dogs. However, limited studies are available with current anti- Leishmania commercial vaccines. A further Plenary Lecture 3.0 Leishmaniasis, limitation of some commercial vaccines Leishvet and One Health includes interference with serological diagnostic methods as well as side effects. July 9, 2019, 8:30 - 10:30 Vaccination may be considered as a part of a Plenary Hall, Madison Ballroom (ABCD), Level 4 comprehensive control program for CanL, in which repellents against sand flies are used concurrently with dog vaccination. Finally, PL03.01 Dog Vaccination to Decrease leishmaniosis is one of the best examples of a Transmission of Parasitic Zoonoses: Notes disease whose successful control depend on from Leishmaniosis the use of a One Health strategy.

Dr. Gaetano Oliva2, Dr. Laia Solano-Gallego1 1 Dep. Medicina I Cirurgia Animals, Universitat PL03.02 Treatment of Canine Leishmaniosis 2 Autonoma de Barcelona, Spain, Department and Drug Resistance – Implications on the of Veterinary Medicine and Food Production, Transmission of Infection and One Health University of Naples Federico II, Italy Prof. Guadalupe Miro2, Prof. Gad Baneth1 Dogs are the main reservoir for Leishmania 1The Hebrew University of Jerusalem, Rehovot, infantum infection and are an important Israel, 2Universidad Complutense de Madrid, target for public health intervention to Madrid, Spain prevent transmission to people. Vaccination is an important preventative tool against this Leishmania infantum causes a zoonotic infection in humans and in dogs. A protective infection endemic in the Mediterranean Basin, vaccine should provide robust memory T cells the Middle East, central Asia, South and able to mount a recall response at the time Central America. It is also a cause of disease of (re)exposure to parasites and reactivate in dogs in the USA, and countries where there polyfunctional T cell effector functions. The is no known sand-fly transmission due to development of safe, effective, durable vertical transmission, travel and importation and low-cost prophylactic vaccines against of infected dogs. Infected dogs may develop leishmaniasis is still a major challenge. severe clinical disease or remain sub-clinically Despite the efforts, there is still no effective infected but infectious to sandflies. Treatment vaccine licenced for human use. However, of canine leishmaniosis (CanL) is warranted four commercial vaccines have been licensed when clinical disease is present, however, for the control of canine leishmaniosis there are very few available effective drugs (CanL) in Brazil and Europe. These vaccines and most of them are used for treatment of provide at least 12 months of protection human disease with the possibility of drug with the indication to reduce the risk of resistance development. This situation calls developing an active infection and clinical for coordination between veterinary and disease. The main controversy regarding human health under the umbrella of One CanL vaccines is that they do not block the Health to decrease the risk of drug resistance establishment of infection. This controversy transmission. has been poorly explored due to difficulty of xenodiagnosis in vaccinated dogs to evaluate The main drugs employed for the treatment transmission. Easy biomarkers to determine of CanL caused by L. infantum are the dogs who are “super-spreaders” of infection pentavalent antimony meglumine antimoniate,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 10 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

miltefosine, and allopurinol. Additional mount a sterilizing immune response predicts immune potentiating compounds such which hosts could be reservoirs of different as domperidone, dietary nucleotides and Leishmania spp. Materials in addition to active hexose have been licensed for dogs parasites are expelled during sand fly during in some countries as ancillary treatment feeding, including salivary antigens and other to enhance immune responses against factors that promote local inflammatory L. infantum infection. Anti-leishmanial responses and visceralization of infection treatment often achieves clinical cure in dogs increasing the likelihood that systemic with leishmaniosis but it is frequently not infection is established. Any environmental associated with a parasitological cure. Treated factor that increases sand fly biting of a dogs may remain carriers of the disease, particular host increases that host’s role in experience clinical relapses and can be Leishmania transmission. infectious to sand flies. First descriptions of reservoir species were Drug resistance has been widely described in based on association with local human human cutaneous and visceral leishmaniosis disease and ability to observe infected but reports on drug resistance in CanL are leukocytes on cytology. This approach relatively scarce. Resistance to allopurinol was one pathogen for one reservoir host. may develop in dogs experiencing disease Advances in sensitive molecular tools greatly relapse which may transmit resistant parasites increased the breadth of mammals found to other dogs and also enhance the danger of to host Leishmania infection. Visceralizing the parasite’s transmission to humans. forms of Leishmania, particularly L. infantum, In conclusion, new treatment and disease are now known to have multiple mammalian control strategies should be coordinated hosts. L. donovani, long been described between human and veterinary officials in a as an anthroponotic parasite, was recently One Health manner to improve the prevention identified through molecular and serologic of disease transmission. surveys to have additional mammalian hosts. The epidemiological role of these animals as a source of parasites to additional hosts via PL03.03 Treatment of Canine Leishmaniosis vector transmission is not known. and Drug Resistance – Implications on the Transmission of Infection and One Health Current evidence may suggest that dogs and other domestic animals may either control Patrick Bourdeau2, Christine Petersen2,3,4 infection or do not have sufficient skin 1Laboratoire de Dermatologie, Parasitologie et parasitemia to be a source of L. donovani to Mycologie, ONIRIS, Ecole National Veterinaire, P. argentipes. Further xenodiagnosis and France, 2Department of Epidemiology, College characterization of skin parasitemia in these of Public Health United States, 3Center for different hosts is required to more broadly Emerging Infections Diseases, United States, understand which Leishmania hosts can be 4IImmunology Program, Department of Internal a source of parasites to sand flies and which Medicine and Microbiology, Carver College of ones are dead-end hosts. Medicine, University of Iowa, United States

Leishmania has biologically adapted to specific Phlebotomine sand flies through long co-evolution. The ability of Leishmania spp. to bind to sand fly midgut allows each Leishmania species to propagate and differentiate into infectious promastigotes and be transmitted. Sand fly feeding upon a mammalian host is the first step towards being infected and a host of Leishmania. Once deposited into the skin, host susceptibility to infection vs. ability to

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 11 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

Plenary Lecture 4.0 Veterinary PL04.02 Discovery, Surveillance, and Control Entomology: Outbreak and of Haemaphysalis longicornis, the Asian Advancements Longhorned tick, in New Jersey

July 10, 2019, 8:30 - 10:30 Dr. Nicole Lewis1 1 Plenary Hall, Madison Ballroom (ABCD), Level 4 NJ Department of Agriculture, Ewing, United States

PL04.01 Introduction to Veterinary On November 7, 2017, Haemaphysalis Entomology: Outbreak and Advancements longicornis, also known as the Asian Longhorned tick, was confirmed by the Prof. Mason Reichard1 National Veterinary Services Laboratory 1Oklahoma State University, United States (NVSL) on a sheep in New Jersey. This was the first finding of the tick infesting livestock in the country. Known as a serious pest of livestock in the Australasian and Western Pacific Regions, this species has never previously established a population in the United States, that we knew of. In a multi-agency effort, this finding led to the identification of H. longicornis in several counties in New Jersey as well as eight other states. The investigation found the tick on several species including dogs, cats, cows, goats, sheep, white-tailed deer, opossum, grey foxes, coyotes, groundhogs, raccoons, horses, and humans. Testing of both the tick and the index case animal, a sheep, were negative for various human and animal pathogens. Currently, state, federal and academic organizations are performing surveillance to better understand the spread of this tick and to help determine the best methods for control. The involved agencies developed educational and outreach material that is available to the public at the NJ Department of Agriculture website. The public is encouraged to submit ticks for identification and a county drop of location has been established for each of the 21 counties. Research is ongoing to better understand disease transmission, pesticide resistance, and spread of this pest throughout the country.

PL04.03 One Health Approach to Manage the Threat of Ticks and Tick-borne Diseases in North America

Dr. Adalberto Pérez de León1 1USDA-ARS Knipling-Bushland U.S. Livestock Insects Research Laboratory & Veterinary Pest Genomics Center, Kerrville, United States

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 12 63rd Annual Livestock Insect Workers Conference PLENARY LECTURES

Ticks are more important than mosquitoes as PL04.04 Ecology and Behavior of vectors of pathogens causing morbidity and Triatomines and Trypanosoma Cruzi at mortality in domestic animals and wildlife. the Human-Wildlife-Dog Interface in the Around 80% of the cattle in tropical and Southern Unites States subtropical regions of the world are affected by economically important ticks and tick- Dr. Sarah Hamer1 borne pathogens. Most tick-borne diseases 1Texas A&M University, College Station, United of public health importance are zoonotic. States Estimates indicate that Lyme disease and other diseases caused by tick-borne Despite over 100 years of research on pathogens could burden over 30% of the Trypanosoma cruzi, this parasite continues global human population by 2050. Some tick to cause significant morbidity and mortality species are invasive and transmit pathogens in human and animal populations across the causing transboundary diseases of high Americas. Parasite transmission cycles are vastly consequence for domestic animals, humans, different across geographic regions, ranging and wildlife. Intricacies of the livestock- from domestic infestations with high human wildlife-human interface represent another contact to sylvatic cycles among wildlife with aspect of global change adding complexity to limited spillover to humans. Using a one health efforts trying to solve the problem with ticks approach, we developed a multidisciplinary and tick-borne diseases (TTBD). “One Health” program to study Chagas disease ecology is a term used to describe approaches to in the southern US, where there is increasing optimize health outcomes for humans, other recognition for locally-acquired disease in both animals, and the environment. Strategies humans and dogs. We use citizen science to based on the One Health concept facilitate empower the public with disease prevention research on the interplay between climate, information while accepting citizen-collected habitat, and hosts driving tick population triatomines. Analysis of over 5,000 triatomines dynamics. This enhances our epidemiological submitted to this program from 27 states since understanding of tick-borne diseases. 2013 revealed >55% T. cruzi infection prevalence Applying the One Health concept provides with two genetic variants of the parasite. Peak a holistic framework to address the global vector encounters with humans occurred in mid- importance of TTBD. Effective and safe summer months, and vector bloodmeals were TTBD management requires rational tactics acquired from diverse wildlife, domestic animals, involving multiple technologies to avoid the and humans. We developed triatomine colonies intense use of chemical treatments, which of locally-relevant species for use in behavioral can harm the environment and public health. and transmission studies. Using time lapse Examples of One Health research to manage photography in our colony as well as in infested the threat of TTBD emphasizing integrated dog kennels, we found that triatomine activity, tick management through the optimal use and therefore risk of host contact, is not limited of compatible methods in a way that is safe, to the nocturnal hours. Further, using an indirect economically viable, and environmentally xenodiagnostic approach, we found variation sustainable will be presented. Our experience in the infectivity of chronically-infected dogs to indicates that grasping the expectations triatomines, in which some dogs consistently of end-users of technology is fundamental infected all naïve bugs to which their blood was to realize a shared vision of improving the fed, whereas others infected fewer insects. In all outcomes of tick control interventions. cases, xenodiagnoses was more sensitive that Scientific evidence can be used to generate PCR of blood to detect host infection. Given the support to establish the capacities the limited antiparasitic treatment options and required for the effective management of lack of a Chagas disease vaccine, disease risk ticks to mitigate the health burden of TTBD. reduction must focus on controlling the vectors in the environment. Citizen science is enabling new insights in vector infection, behavior, and host encounters that can ultimately be useful in developing interventions to protect human and animal health.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 13 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 14 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Oral Sessions

OA01 Canine Heartworm I (a historically endemic area), Oklahoma and Kansas, and Florida. July 8, 2019, 11:00 - 12:30 Plenary Hall, Madison Ballroom (ABCD), Level 4 Conclusions Canine heartworm prevalence is increasing in much of the United States both regionally and OA01.01 Regional and Local Trends in the locally despite veterinarian recommendations Prevalence of Canine Heartworm: United on prevention and testing. Additional steps States 2012–2018 should be taken to protect dogs, cats, and ferrets. Further work is needed to identify the Mrs. Stella Self2, Dr. Cassan Pulaski3, Dr. driving factors of the local negative trends Christopher McMahan2, Dr. D. Andrew Brown2, present along the Mississippi Alluvial plain, Dr. Michael Yabsley1, Dr. Jenna Gettings1 Florida, and other areas. 1University Of Georgia, Athens, United States, 2Clemson University, Clemson, United States, 3Louisiana State University, Baton Rouge, OA01.02 A Moxidectin Extended-Release United States Injectable Suspension (PROHEART® 12) for the Prevention of Heartworm (DIROFILARIA Aims IMMITIS) Disease in Dogs in the USA for 12 Canine heartworm disease is a potentially Months fatal disease for which treatment is financially burdensome for many pet owners. Prevention Dr. Tom McTier1, Ms. Martha Wachowski1, is strongly advocated by the veterinary Dr. Kristina Kryda1, Mr. Sean Mahabir1, Dr. community along with routine testing for Doug Rugg1, Dr. Mark Mazaleski1, Dr. Dwight infection during annual wellness examinations. Bowman2 Despite the availability of efficacious 1Zoetis, Kalamazoo, United States, 2Cornell chemoprophylaxis, recent reports have University, Ithaca, United States suggested that the incidence of heartworm disease in domestic dogs is increasing. ProHeart® 6 (PH 6) (0.17 mg/kg moxidectin) Identification of areas of increasing prevalence and ProHeart® SR-12 (PH 12) (0.5 mg/ is imperative for effective client education, kg moxidectin) are extended-release intervention efforts, and focusing research. microsphere injectable products providing continuous heartworm prevention for 6 and Results 12 months, respectively, in dogs. Over 12.4 Using heartworm testing data for the United million PH 12 doses have been sold globally States from January 2012 through September outside the United States (USA) since 2008. 2018, a Bayesian spatio-temporal binomial The current studies were conducted to regression model was used to estimate confirm the efficacy of PH 12 for preventing the regional and local temporal trends of the development of Dirofilaria immitis in dogs heartworm prevalence. The region with the in the USA. Two separate laboratory studies greatest increase in prevalence was found in were conducted, each with 20 Beagles, the Lower Mississippi River Valley. Statistically ≥12 months of age, of mixed sex, weighing significant increases in prevalence occurred between 6.7 and 12.5 kg and determined to throughout the southeastern states and be free of existing heartworm infection prior extended northward into Illinois and Indiana. to study initiation. Dogs were then randomly Local trends varied across the United States, allocated to receive a single subcutaneous with positive trends along most of the Atlantic injection of either saline (control) or PH coast, central United States, and western 12 on Day 0. PH 12 was reconstituted to states, while clusters of negative trends were deliver moxidectin at 0.5 mg/kg. Dogs were present along the Mississippi Alluvial Plain inoculated with 50 D. immitis third stage

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 15 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

larvae on Day 365. Two different heartworm of fluralaner was identified as the longer isolates (GCFL and ZoeAL) collected from dosing period and the next most frequent naturally infected dogs in the USA were benefit was convenience. The proportional used. All dogs were necropsied ~5 months owner responses were surprisingly consistent post-inoculation, and examined for adult across the three countries. Several questions heartworms. In both studies, none of the PH compared 12-week dosing with monthly 12-treated dogs had any adult heartworms, dosing to assess whether the longer dosing whereas all control dogs in both studies had was a burden or more difficult to accomplish. heartworms, with a geometric mean of 30.2 Most responders indicated that they did not (range, 22-37) for Study 1 and 32.6 (range, have difficulty providing follow-up doses and 22-44) for Study 2. Mean counts for PH 12 that they were more likely to dose fluralaner for both studies were significantly different on time compared to monthly products. from mean counts for comparable controls (p<0.0112). One PH 12-treated dog and two saline-treated controls had mild injection site OA01.04 Macrocyclic Lactone Resistance reactions within 24 hours post-treatment that in Dirofilaria immitis: Extent of Resistance resolved within one month. In conclusion, a and Investigation of the Genomic Changes single dose of PH 12 (0.5 mg/kg moxidectin) Underlying Resistance was 100% effective in preventing heartworm (D. immitis) disease in dogs for 12 months Prof. Roger Prichard1, Dr Cristina Ballesteros1, in two laboratory studies using recently Prof Timothy Geary1, Ms Kathy Keller1, Dr collected heartworm isolates from the USA. Catherine Bourguinat1, Dr Cassan Pulaski2 1McGill University, Sainte Anne-de-Bellevue, Canada, 2Louisiana State University, Baton OA01.03 A Comparison of Dog Owner Rouge, United States Satisfaction and Preference for Flea and Tick Medications in the US, UK and Australia Prophylaxis with macrocyclic lactone (ML) endectocides is the primary strategy for Dr. Robert Lavan1, Dr. Rob Armstrong1, Dr. heartworm control. Recent evidence has Dorothy Normile1, Hannah Newbury1, Karen confirmed that ML-resistant Dirofilaria Lipworth1 immitis isolates have evolved. In this study, 1Merck Animal Health, Perkasie, United States participating veterinary clinics helped recruit heartworm positive dogs. Blood was taken A dog owner survey was administered to prior to treatment and then positive dogs over 1500 dog owners in the US, UK and were treated with a single standard dose Australia to assess their opinions on an of Advantage Multi® and another blood extended duration flea and tick medication sample taken 2–4 weeks post-treatment. A (Bravecto®[fluralaner], Merck Animal Health) modified Knott’s Test was undertaken on and compare this experience to using monthly each blood sample, followed by isolation of flea and tick products. Additionally, at least microfilariae, genomic DNA extraction and one veterinarian in each clinic provided their MiSeq sequencing of regions encompassing annual flea and tick recommendation. 10 SNP sites correlated with ML resistance. We observed significant correlation of SNP While veterinarians, on average, loci frequencies with the ML microfilaricidal recommended 12 months protection for fleas response phenotype. A 2-SNP model and 10-12 months for ticks, dog owners usually was superior to other models tested. believed that they needed less protection Subsequently, we have undertaken whole for their dogs. All dog owners were current genome sequencing on a number of the users of the extended duration fluralaner isolates that are phenotypically ML- sensitive and most dog owners (75%) had used other (S) or ML-resistant (R). Fst values (an index of monthly flea and tick products. Satisfaction identity for which an Fst = 0 means identity, was very high among respondents (over and an Fst = 1 means complete difference) of 90%) and most (over 90%) preferred the the S and R isolates have been determined extended duration product. The chief benefit and single nucleotide polymorphisms (SNPs)

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 16 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

in which the S versus R Fst ≥ 0.5 have been purchased monthly topical or oral products identified. Sequences with high Fst values were significantly more likely to purchase were blasted against the Brugia malayi (a 1-6 months of protection relative to the filarial nematode phylogenetically very close extended duration flea/tick product. Owners to D. immitis) and the Caenorhabditis elegans who purchased the longer acting product genomes to identify orthologous genes in were significantly more likely to purchase D. immitis and to map them to the B. malayi 7-12 months of protection and thereby come genome. These comparisons revealed that closest to adhering with veterinary flea/tick a high proportion of the genetic differences control recommendations. Regardless of between S and R isolates map to a region of flea/tick product prescribed or dispensed, one of the five chromosomes in D. immitis; veterinarians have a lot of room to improve genes in that region may contribute to ML owner adherence for flea and tick protection. resistance. Work is continuing to define more precisely the genetic change(s) responsible for ML resistance and to understand the underlying OA01.06 Functional Expression of mechanism of resistance in heartworm. Glutamate-Gated Chloride Channels from the Parasitic Nematode Brugia Malayi

OA01.05 Compliance With Veterinary Melanie Abongwa1, Bukunmi Akanji1, Sudhanva Recommendations for Canine Flea and Tick Kashyap1, Aaron Hoisington1, Shivani Medications in Spain and the United States Choudhary1, Saurabh Verma1, Gunnar Mair1, Richard Martin1, Alan Robertson1 Dr. Robert Lavan1, Dr. Federica Burgio2, Dr. 1Iowa State University, Ames, United States Kaan Tunceli3, Dr. Dorothy Normile1 1Merck Animal Health, Perkasie, United States, Filarial nematodes cause significant disease in 2MSD Animal Health, Zaragosa, Spain, 3Merck, humans and animals. Control and treatment Inc., Kenilworth, United States are severely hampered by the lack of drugs available to kill adult worms. Current therapies Improvements in patient health outcomes are are only effective against larval microfilariae, associated with development of longer duration with limited activity against adult worms. medications for chronic diseases and chronic Therefore, there is a need to develop safe administration. In 2014, a longer duration and effective adulticides. The elucidation isoxazoline product was launched to treat or of the pharmacology of potential drug prevent ectoparasite infestation on dogs. This targets is a key element in reaching this goal. product (Bravecto® [fluralaner]; Merck Animal Glutamate-gated chloride channels (GluCls) Health) is an oral chew that is effective for up are targets for the macrocyclic lactone class to 12 weeks. Most other available topical or oral of anthelmintics which include the ‘wonder flea/tick products are re-dosed monthly. drug’, ivermectin. Here, we report on the cloning and functional characterization in Studies were completed on large databases Xenopus oocytes of two B. malayi GluCls; in Spain and the United States to look Bma-GLC-3 and Bma-AVR-14a. Functional, at dog owner adherence to veterinary homomeric channels were formed when recommendations around fleas and ticks. oocytes were injected with either glc-3 or The studies demonstrated that, regardless of avr-14a capped RNA (cRNA). Two-electrode flea/tick medication brand, 50-60% of dog voltage-clamp recordings revealed significant owners purchased only 1-2 doses per year. pharmacological differences between the Also, owners who purchased the extended channels formed by GLC-3 and AVR-14a. duration flea and tick product obtained GLC-3 was more sensitive to L-glutamate significantly more months of coverage per compared to AVR-14a; EC50 values were 64.8 year than owners who purchased monthly and 1.6 mM, respectively. Current-voltage products. In both countries, owners who relationships demonstrated that the channels purchased a topical product purchased the wereµM selectively permeable to chloride ions. least number of annual doses, on average, No responses were detected following the compared to oral products. Dog owners who application of 1 mM L-aspartate, glycine,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 17 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

-aminobutyric acid (GABA), -amino-3- address these distinctions. These issues hydroxy-5-methyl-4-isoxazolepropionic acid highlight the necessity for developing new (AMPA),γ or N-methyl-D-aspartateα (NMDA). World Association for the Advancement of AVR-14a was also more sensitive to the Veterinary Parasitology (WAAVP) guidelines chloride channel antagonists picrotoxin and that improve both the general methodology fipronil. Ivermectin exhibited an unusual and and standardization of the FECRT. Extensive complex mode of action on GLC-3 involving data simulations and analyses identified direct activation of the channel (EC50 = many factors that can affect the outcome 4.1 nM) and inhibition of the responses and interpretation of a FECRT. One important to L-glutamate (IC50 = 95.1 pM). Further factor is the level of over-dispersion in the investigation of these subunits and the in pre-treatment FEC on a given farm (k). While vivo subunit composition and functions of k is unknown before FEC are measured, k is the different GluCls present in B. malayi known to differ among hosts and age groups, will provide insights to ivermectin’s limited and this heavily impacts the amount of data activity on adult filaria. necessary to have sufficient power to make a correct interpretation of the observed results. Proper consideration of this and other OA02 Diagnosis and Decision Support factors made it obvious that a simple protocol for GI Nematodes in Ruminants I would be unlikely to yield consistently accurate results. However, there is a strong July 8, 2019, 11:00 - 12:30 demand among the veterinary parasitology Breakout Room 2, Hall of Ideas E&H, Level 4 community for a simplified guideline that will facilitate the performance of FECRT at the individual farm level. To address these OA02.01 Further Progress on Developing New conflicting demands, we have developed a World Association for the Advancement of guideline that has several components. In Veterinary Parasitology (WAAVP) Guidelines the first part, we address the major issues for the Fecal Egg Count Reduction Test relevant to experimental design and make a series of general recommendations. We Ray Kaplan1, Bruno Levecke2, Paul Torgerson3, then provide two separate guidelines for Robert Dobson4, Stig Thamsborg5, Matt the performance of the FECRT, (1) a more Denwood5, Martin Nielsen6, Jozef Vercruysse2 rigorous version that is intended for use in 1Department of Infectious Diseases, College scientifically-based studies (e.g. in studies of Veterinary Medicine, University of Georgia, intended for publication or registration of Athens, United States, 2Department of new drugs), and (2) a simpler version that has Virology, Parasitology and Immunology, fewer experimental and analytical demands, Faculty of Veterinary Medicine, Ghent and is intended for use by veterinarians and University, Ghent, Belgium, 3Section of livestock owners. Epidemiology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland , 4School of Veterinary and Life Sciences, Murdoch OA02.02 Calf Behaviour as a Targeted University, Perth, Australia, 5Department of Selective Treatment (TST) Indicator Veterinary and Animal Sciences, University of Copenhagen, Copenhagen, Denmark, 6Maxwell Dr. Andrew Greer1, Dr. Rachael Bryant1, Mr. H. Gluck Equine Research Center, University Robin McAnulty1 of Kentucky, Lexington, United States 1Lincoln University, Christchurch, New Zealand

In recent years there have been many new Targeted selective anthelmintic treatments insights into the fecal egg count reduction (TSTs) rely on being able to accurately identify test (FECRT) regarding optimal experimental individuals within a herd or mob that are likely design and the analysis and interpretation of to benefit from receiving an anthelmintic data. Additionally, there are important host- treatment. Given that one of the ubiquitous specific and parasite-specific differences signs of sub-clinical parasitism in grazing that require protocol modifications to livestock is a reduction in voluntary feed

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 18 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

intake, the objective of this pilot study was during ~140 days. At turnout, experimental to evaluate whether grazing and/or activity animals were allocated to one of two replicate behaviour could be used as a TST indicator treatment groups (H1; H2; L1; L2) grazing in grazing calves. Twenty-five 10-month-old in different enclosures. Infected groups (H) Friesian x Jersey calves naturally infected with received 10.000 third stage (L3) O. ostertagi gastro-intestinal nematodes, and with faecal (50%) and C. onchophora (50%) larvae; egg counts ranging between 0 and 250 eggs whereas control groups (L) were dewormed per g, were fitted with Sensoor ™ activity and monthly with 0.5 mg ivermectin (Ivomec®, behaviour ear tags which record grazing time, Pour-on) per kg bodyweight. Activity and ruminating time and activity. On one of four rumination patterns were monitored by fitting treatment days each animal was administered animals in each group (H1, n=15; H2, n=17; L1, anthelmintic and their grazing and activity n=17; L2, n=11) with leg mounted (IceQube, pre and post treatment were compared. IceRobotics) and neck mounted (Heatime LD Grazing time initially decreased from a mean tag, SCR) loggers. Body weight gain (BWG) of 322 ± 23.9 minutes per day pre-treatment was recorded every fortnight, whereas faecal to 223 ± 14.6 minutes per day in the 24h and blood samples were collected every immediately following treatment (P=0.04) four weeks for nematode faecal egg count and then increased to 422 ± 27.1 minutes per (EPG) and serum pepsinogen concentrations. day post treatment (P=0.003, compared with EPG levels were affected by treatment (P pre-treatment). Rumination time also initially < 0.0001). However, BWG was not affected decreased from a mean of 288 ± 21.0 minutes (P = 0.997) by treatment, probably due to per day pre-treatment to 95 ± 4.6 minutes extreme drought. Preliminary results shows an per day in the 24h immediately following increase in lying time (P = 0.0369) recorded treatment (P=0.001) and then increased to by IceQube loggers and a trend of increase in 362 ± 18.8 minutes per day post treatment individual behaviour variation for rumination (P=0.01, compared with pre-treatment). There (P = 0.0556) and decrease in individual were no differences in the time spent being behaviour variation for activity (P = 0.0654) in active (P=0.71), not active (P=0.41) or highly infected groups, recorded by Heatime loggers active (P=0.14). Overall, these results indicate during the 30 first days. In addition, a trend that both grazing time and rumination time, of an increase in total step count (P = 0.0565) but not activity time, respond to anthelmintic and activity (P = 0.0509) in infected groups, treatment and may provide a useful indicator recorded by IceQube loggers over ~140 days, for treatment as part of a TST regime. was seen. In conclusion, our preliminary data supports that changes and variation in activity and rumination patterns monitored with OA02.03 Effects of Nematode Parasitism on loggers could contribute to the identification Activity Patterns and Rumination Behaviour of animals challenged with GIN, even when no in First-Season Grazing Cattle differences can be observed with traditional measures, such as BWG. Niclas Högberg1, Professor Lena Lidfors2, Associate professor Anna Hessle2, Professor Johan Höglund1 OA02.04 Construction of a Decision Tree for 1Swedish University of Agricultural Sciences, Targeted-Selective Treatment of Dairy Cows Department of Biomedical Sciences and Against Gastrointestinal Nematodes Veterinary Public Health; Parasitology Unit, Uppsala, Sweden, 2Swedish University of Nadine Ravinet1, Anne Lehébel1, Nadine Agricultural Sciences, Department of Animal Brisseau1, Yann Quenet1, Nathalie Menudier2, Environment and Health, Skara, Sweden Aurélien Madouasse1, Christophe Chartier1, Alain Chauvin1 We investigated the effects of gastrointestinal 1BIOEPAR, INRA, Oniris, Nantes, France, nematode (GIN) challenge on activity and 2CEVA Santé Animale, Libourne, France rumination patterns in first season grazing In dairy cows, the milk production (MP) steers exposed to two different levels of response to anthelmintic treatment (AT) Ostertagia ostertagi and Cooperia oncophora against gastrointestinal nematodes is highly

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 19 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

variable. This study aimed to develop a OA02.05 Ostertagia Ostertagi Antibodies decision tree based on nested criteria to in Bulk Tank Milk in Autumn: Variability of increase the probability of MP response. the ELISA Results According to the Date of Sampling A randomized controlled trial was conducted using injectable eprinomectin at housing Nadine Ravinet1, Nadine Brisseau1, Anne in 123 French grazing dairy herds. Monthly Lehébel1, Yann Quenet1, Nathalie Menudier2, individual milk yields were obtained. Linear Aurélien Madouasse1, Christophe Chartier1, mixed models were used to assess the effect Alain Chauvin1 of AT on MP. Starting with all herds (Step 0), 1BIOEPAR, INRA, Oniris, Nantes, France, the herd level criterion associated with the 2CEVA Santé Animale, Libourne, France highest MP response to AT was identified (step 1). Then, among farms meeting this In dairy cows, the bulk tank milk (BTM) first criteria, a second criteria associated Ostertagia antibody level (ELISA, results with the highest MP response was identified as optical density ratio, ODR) is often used (step 2). This was repeated a third time on as a criterion for anthelminthic treatment herds meeting the first 2 criteria (step 3). at housing, one single BTM sample being Furthermore, at each step, within the selected collected between October and December. herds, the cow characteristics associated with However, this autumnal BTM ODR could vary the highest MP gain were identified. depending on the date of sampling. The objective of the study was to quantify the When considering all the herds (step 0), the autumnal variability in BTM Ostertagia ODRs, effect of AT was small but significant (+0.3 taking into account the variability related to kg/cow/day). The first criterion (step 1) was the measurement method. the % of grazed grass in the diet: no MP gain in low-pasturing herds versus +1 kg/cow/day Ostertagia antibody levels were measured in moderate/high-pasturing herds. In this 16 times in 6 BTM samples (O. ostertagi-Ab subset (n=50), the second criterion (step 2) ELISA kit, SVANOVIR®) (4 days, 2 experiments was the TEC (Time of Effective Contact with per day, samples tested twice per experiment, GIN infective larvae before the first calving): same operator). The coefficient of variation +0.6 kg/cow/day in high-TEC herds versus (CV), calculated for each series of 16 ODRs, +1.4 kg/cow/day in low-TEC herds. In this ranged from 0.08 to 0.10. new subset (n=17), the third criteria (step 3) was the bulk tank milk Ostertagia ODR: In 125 French grazing dairy herds, BTM +0.2 versus +1.8 kg/cow/day when ODR < 0.9 samples were collected fortnightly from the and ≥ 0.9, respectively. In steps 0, 1 and 2, 1st of October until housing and analysed best responding cows had calved during the with the same ELISA kit: 3 to 7 autumnal grazing season and/or were low-producing ODRs per herd were obtained. The effect of cows within their herd. the sampling date on the ODR was assessed (linear mixed model, herd as random effect) Those herd and cow-level criteria, used and the CV of each within-herd series was sequentially, can be useful to build a decision calculated. tree that increases the probability of MP gains. November and December ODRs were slightly lower than October ODRs : -0.03 and -0.07, respectively (p =0.0002). The maximal ODRs were obtained during October in 99/125 herds (79%). Within-herd CVs ranged from 0.02 to 0.38 (mean 0.10) and 44 herds (35%) showed a CV > 0.10, indicating that the autumnal variation in BTM ODR values could be higher than the variability related to the method of measurement.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 20 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Those results confirm that the BTM Ostertagia 0.755, P<0.01). These results together suggest antibody level can vary during the autumn that the productivity of Australian dairy farms and this variability can have consequences on has the potential to improve with better the decision to treat. To minimize the risk to animal health management. under-estimate the exposure to Ostertagia, the optimal month for sampling BTM should be October. OA03 North American Ticks

July 8, 2019, 11:00 - 12:30 OA02.06 A Bulk Milk Tank Survey of Breakout Room 3, Hall of Ideas F&I, Level 4 Ostertagia and Fasciola Hepatica Antibodies in Australian Dairy Enterprises. OA03.01 Show Us Your Ticks: A Survey of Ticks Infesting Dogs and Cats Across the Dr Gareth Kelly1, Grant Richards2, David United States Homer3 1Boehringer Ingelheim, Sydney, Australia, Dr. Meriam Saleh1, Kellee Sundstrom1, Dr. 2Parasite Diagnostic Services, Hastings, Kathryn Duncan1, Michelle Ientile1, Julia Jordy1, Australia, 3Landmark, Melbourne, Australia Parna Ghosh1, Dr. Susan Little1 1Oklahoma State University, Stillwater, United Between 2010 and 2016, 2493 bulk milk States samples were analysed to determine the prevalence of Ostertagia and Liver Fluke A variety of tick species infest dogs and cats antibodies (measured as Optical Density in the United States. To identify the species Ratios (ODR)) to help inform farmers on the and stages of ticks on pets and characterize health status of their milking herds. Samples host attachment site preferences, tick from herds with no exposure to anthelmintic submissions were invited from veterinary treatment within the previous four months clinics in all 50 states. Upon receipt, ticks were submitted to a single laboratory by sales were identified to species and stage using managers. Results were collated according morphologic keys; if damage precluded to the region covered by the sales manager, identification by morphology, species was closely approximating biogeographical confirmed molecularly. From February 2018 zones of Australia and enabling samples to to January 2019, 10,885 ticks were submitted be categorized according to the major dairy from 1,487 dogs and 341 cats in 49 states; producing regions within Australia. Overall, ticks were collected in every month. Dog and the average ODR for Ostertagia was 0.788, cat infestation intensities ranged from 1-4,765 indicative of a moderate infection (higher than and 1-52 (median = 1, mean = 6.7 and 2.8), published surveys where year-round grazing respectively. Dogs were primarily infested is not routine). Based on previous published with Dermacentor variabilis (527/1487; validation work in other countries, this level 35.4%), Ixodes scapularis (400/1487; 26.9%), of immune response to Ostertagia challenge Amblyomma americanum (348/1487; 23.5%), in dairy cows equates to an estimated milk and Rhipicephalus sanguineus (172/1509; loss of 0.9 kg/day/cow. Furthermore, 92% of 11.6%). Cats were primarily infested with I. herds indicated that productivity may have scapularis (156/341; 45.7%), A. americanum been reduced with an ODR greater than (100/341; 29.3%), and D. variabilis (62/341; 0.501 when tested. No seasonal variation was 18.2%). Other submitted ticks included A. evident, but regional variation was evident maculatum, Haemaphysalis longicornis, with lower ODR values in the sub-tropical Otobius megnini, and less common region of NSW compared to temperate dairy Dermacentor spp. and Ixodes spp. Co- production regions. For fluke, 38% of all farms infestations were documented in 93 dogs tested positive with moderate to high ODRs. and 14 cats. Reported attachment site by tick Herds with high fluke ODR levels also had species differed significantly for both dogs significantly higher ODR levels for Osteratgia ( 2 = 221.213, df = 15, p < 0.0001) and cats ( 2 than herds with low fluke ODRs (0.851 vs = 244.689, df = 10, p < 0.0001). In dogs, A. χ χ

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 21 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

americanum was most commonly attached to dogs and cats. Together, these data support the abdomen, axillary, and inguinal regions, the interpretation that D. variabilis is the D. variabilis and I. scapularis the head and predominant Dermacentor species found on ears, and R. sanguineus head, ears, legs, and dogs and cats throughout the United States, feet. In cats, D. variabilis and I. scapularis were a finding that may reflect somewhat recent most commonly attached to the head and shifts in tick distribution. ears, but A. americanum was most commonly attached to the tail and perianal region. These data confirm that dogs and cats in the United OA03.03 Ixodes spp. from Pet Dogs and Cats States are at risk of tick infestation every in the United States: Geographic Distribution month of the year, and that tick species differ and Prevalence of Borrelia burgdorferi in their reported attachment sites. Ms. Parna Ghosh1, Dr. Meriam Saleh1, Ms. Kellee Sundstrom1, Ms. Michelle Ientile1, Dr. Susan OA03.02 Dermacentor spp. from Dogs Little1 and Cats in North America: Diversity and 1Center for Veterinary Health Sciences, Geographic Distribution Oklahoma State University, Stillwater, United States Kathryn Duncan1, Meriam Saleh1, Kellee Sundstrom1, Susan Little1 Ixodes spp. are commonly found on dogs 1Oklahoma State University, Stillwater, United and cats in many areas of the world. In States the eastern United States, I. scapularis, the predominant species and primary vector of Dermacentor spp. ticks are known to commonly Borrelia burgdorferi, can be divided into two feed on dogs and cats throughout North genotypes – American and Southern – on the America and transmit important tick-borne basis of 16S rRNA gene sequence. To confirm pathogens, including spotted fever group the species and characterize the genotype Rickettsia spp. However, the geographic of Ixodes ticks submitted from dogs and distribution of the different species of cats, we examined ticks morphologically Dermacentor in this region is not completely and, when indicated, evaluated a 16S rDNA understood. To better define the identity and sequence from, 342 ticks submitted from distribution of Dermacentor spp. removed 185 dogs, 63 cats, and one rabbit from 34 from dogs and cats in the United States, 495 states; to estimate prevalence of infection, Dermacentor spp. ticks submitted from 260 flaB of B. burgdorferi sensu stricto was dogs (n=449 ticks) and 39 cats (n=46 ticks) amplified from a subset of ticks. Almost all from veterinary practices in 40/50 states were Ixodes spp. submitted from the Northeast identified morphologically and molecularly. (n=93/94; 98.9%) and Midwest (n=55/57; Amplification and sequencing of the ITS2 region 98.2%) were I. scapularis, American genotype and a 16S rRNA gene fragment confirmed that (primary haplotype F), and 26/140 (18.6%) 97.8% (484/495) were D. variabilis and 1.6% were positive for B. burgdorferi. One I. affinis (8/495) were D. albipictus. In total, only three was identified from a shelter cat in New York, (0.6%) D. andersoni were identified, one each one I. cookei from a dog in Ohio, and one from one cat and one dog from Montana and I. scapularis Southern genotype (O) from a one cat from Idaho. While translocation of pets dog in Kansas. In contrast, 77/144 (53.5%) prior to tick removal cannot be discounted, Ixodes spp. from the South were I. scapularis, the majority (36/45; 80%) of Dermacentor American genotype (F and K); 50/144 (34.7%) spp. ticks removed from dogs and cats in the were I. scapularis, Southern genotype (M, Rocky Mountain states, where D. andersoni N, and O); 7/144 (4.9%) were I. affinis; and has been thought to predominate, were 10/144 (6.9%) were I. cookei. None of the actually D. variabilis, suggesting this species Ixodes spp. from the South tested positive may be more widespread in the western for B. burgdorferi. In the West, most (40/47; United States than is currently recognized, or 85.1%) Ixodes spp. were I. pacificus, with I. that D. andersoni, if also present in the region, angustus (n=3) submitted from dogs in Alaska is preferentially feeding on hosts other than and Oregon and I. haerlei (n=4), preliminarily

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 22 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

identified from a dog in Montana. Although I. H. longicornis using molecular techniques, our scapularis, American genotype predominated objective was to characterize the vssc gene. in the Northeast, a diverse array of Ixodes spp. RNA was obtained from a pool of H. longicornis were found on dogs and cats in the South and collected in NJ. Degenerate primers were used West; the significance of these less common to amplify two conserved segments of the vssc Ixodes spp. as disease vectors, if any, warrants gene and a 3’/5’ rapid amplification of cDNA further investigation. ends approach was used to get the full-length transcript. The translated amino acid sequences and the putative protein secondary structure OA03.04 Characterization Of Voltage Sensitive was compared to the homologs vssc of the Sodium Channel Gene Of Haemaphysalis tropical cattle tick, Rhipicephalus microplus longicornis (Acari: Ixodidae), A New Invasive (Canestrini), the brown dog tick, Rhipicephalus Tick Species In The United States sanguineus (Latreille) and other parasitic arthropods. No mutations previously associated Dr. Guilherme Klafke1,2, Jason Tidwell1, Dr. to pyrethroid resistance were detected in the NJ Robert Miller1, Prof. Dina Fonseca3, Dr. Denise H. longicornis samples analyzed. To the best of Bonilla4, Dr. Adalberto Pérez de León5 our knowledge, this is the first characterization 1USDA-ARS Cattle Fever Tick Research of a gene in H. longicornis associated with Laboratory, Edinburg, United States, 2Instituto acaricide resistance. de Pesquisas Veterinarias Desiderio Finamor, Secretaria da Agricultura do Rio Grande do Sul, Eldorado do Sul, Brazil, 3Rutgers Center OA03.05 Dynamic Expression Analysis of for Vector Biology, Rutgers University, New microRNA Profiles During Haemaphysalis Brunswick, United States, 4USDA-APHIS/ Longicornis Different Stages by Deep VS National Cattle Fever Tick Eradication Sequencing of Small RNA Libraries Program, Natural Resources Research Center, Fort Collins, United States, 5USDA-ARS Wenge Liu1, Junhui Guo1, Jin Luo1, Hui Shen1, Knipling-Bushland U.S. Livestock Insects Qiaoyun Ren1, Ze Chen1, Zhiqiang Qu1, Zegong Research Laboratory and Veterinary Pest Wu1, Jun Ni1, Xiaofeng Xu1, Jianxun Luo1, Hong Genomics Center, Kerrville, United States Yin1, Guangyuan Liu1 1Lanzhou Veterinary Research Institute, Haemaphysalis longicornis (Neumann), the Chinese Academy of Agricultural Science, Asian longhorned tick, is a three-host tick Lanzhou, China species that originates from temperate and subtropical areas of East Asia where it infests Haemaphysalis longicornis, a three-host life and transmits zoonotic pathogens to livestock cycle tick, is an obligate specialized blood- and humans. H. longicornis was detected sucking parasite. MicroRNAs (miRNAs) show in the United States outside of quarantine an abundance of expression changes during for the first time on August 2017 infesting the different developmental stages of ticks. sheep in New Jersey (NJ). Since then, this Here, deep sequencing technology was invasive tick has been detected in Arkansas, used to profile the conserved and potential Connecticut, Maryland, New York, North novel miRNAs expressed during different Carolina, Pennsylvania, Virginia, and West developmental stages of H. longicornis. In Virginia. Treatment of livestock with acaricidal total, 111,192,069 reads were obtained and four drugs has been the main strategy to control small RNA libraries (egg, unfed larvae, unfed ticks. However, the development of acaricide nymph and unfed adults) of H. longicornis resistance needs to be anticipated. Acaricide were built after deep sequencing on a resistance is an inherited phenotypic trait Hiseq 4000. Among these, 78 conserved frequently conferred by mutations in the miRNAs and 55 potential novel miRNAs were pesticide’s target sites. Pyrethroid resistance identified, including stage-specific miRNAs in several arthropod species can be caused by and differentially-expressed miRNAs. GO conserved mutations in the voltage sensitive analysis indicated that significantly-enriched sodium channel gene (vssc). In order to detect GO terms related to cell proliferation and future development of pyrethroid resistance in differentiation, comprising the specific terms

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 23 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

developmental process, growth, metabolic from H. longicornis (HlGST and HlGST2) were process, regulation of biological process, identified until now. However, the role of GSTs reproduction and membrane enzyme regular in pyrethroid metabolism of ticks has still activity. KEGG analysis revealed significantly- been unknown. enriched signaling pathways related to growth and development including the [Objective] In this study, we tried to elucidate insulin signaling pathway, the notch signaling the role of GST in flumethrin metabolism pathway, the Hippo signaling pathway and the by determining their possible interaction, Wnt signaling pathway. Our data suggested its ability to induce GST gene and protein that the abundance of miRNA changes (both expression, and effect of GST knockdown on conserved and potential novel) in the different tick survival upon exposure to flumethrin. developmental stages of H. longicornis, especially for stage-specific miRNAs, may [Methods] Enzyme kinetic studies were indicate the importance of miRNAs as conducted with recombinant H. longicornis essential regulators during the development GSTs. Using the Vmax and Km values of H. longicornis. of recombinant GSTs with and without acaricides, the type of interaction between recombinant GSTs and flumethrin was OA03.06 The Haemaphysalis Longicornis determined. The sublethal doses of flumethrin Larvae Needs Glutathione S-Transferase in were determined using a modified larval the Metabolism of Flumethrin packet test. The gene and protein expression of adult female ticks exposed to sublethal Emmanuel Hernandez1,2, Dr. Kodai Kusakisako1, doses of flumethrin were also determined Dr. Melbourne Talactac1,3, Dr. Remil Galay1,4, Dr. using real-time RT-PCR and Western blot Takeshi Hatta5, Dr. Kozo Fujisaki6, Dr. Naotoshi analysis. RNA interference of GST genes was Tsuji5, Dr. Tetsuya Tanaka1,2 conducted on different stages of development 1Laboratory of Infectious Diseases, Joint to determine the effect of GST knockdown on Faculty of Veterinary Medicine, Kagoshima tick survival upon exposure to flumethrin. University, Kagoshima, Japan, 2Department of Pathological and Preventive Veterinary [Results] Flumethrin showed uncompetitive Science, The United Graduate School of inhibition on the activity of recombinant Veterinary Science, Yamaguchi University, HlGST, while recombinant HlGST2 activity was Yoshida, Japan, 3Department of Clinical and not affected. Both GST genes and proteins Population Health, College of Veterinary were upregulated upon exposure to sublethal Medicine and Biomedical Sciences, Cavite doses of flumethrin. Knockdown of GST genes State University, Indang, Philippines, did not affect the survival of the nymphs 4Department of Veterinary Paraclinical and adults, while HlGST knockdown resulted Sciences, University of the Philippines at Los to the increased susceptibility of larvae to Baños, College, Philippines, 5Department sublethal doses of flumethrin. of Parasitology, Kitasato University School of Medicine,, Sagamihara, Japan, 6National [Conclusion] These findings suggest that Agricultural and Food Research Organization, HlGST is vital for the metabolism of flumethrin Tsukuba, Japan in the larvae.

[Introduction] Haemaphysalis longicornis is a tick of health importance as it serves as vector of several disease-causing pathogens. Presently, the major method to control H. longicornis is by using chemical acaricides. Flumethrin is a common pyrethroid pesticide that has specificity towards acarids. Glutathione S-transferases (GST) system on ticks is one of the mechanisms for metabolizing these acaricides. Two GSTs

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 24 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA04 Coccidia proliferation was almost completely inhibited (>95%) at 200 nM. Nonetheless, exposure of July 8, 2019, 11:00 - 12:30 infected cultures to 200 nM BKI 1369 for five Breakout Room 4, Hall of Ideas G&J, Level 4 days did not induce phenotypic morphological alterations in surviving merozoites as confirmed by transmission electron microscopy. During OA04.01 Bumped Kinase Inhibitor 1369 Is treatment in piglets, plasma concentration Effective Against Porcine Cystoisosporosis of BKI 1369 increased to 11.7 M, suggesting constant exposure of parasites to the drug. Oral Aruna Shrestha1, Florian Koston1, Bärbel application of BKI 1369 couldµ be considered as Ruttkowski1, Ryan Choi2, Grant R. Whitman2, a new therapeutic alternative against porcine Samantha A. Michaels2, Edelmar Navaluna2, cystoisosporosis. For the use in pigs, future Kayleigh F. Barret2, Lynn K. Barret2, Carlie S. studies on BKI 1369 should be directed towards Dorr2, Kayode K. Ojo2, Matthew A. Hulverson2, ease of drug handling and minimizing treatment Andrew Hemphill3, Wesley C. Van Voorhis2, frequencies. Anja Joachim1 1Institute of Parasitology, University of Veterinary Medicine Vienna, Vienna, Austria, OA04.02 In Vivo Anti-Cryptosporidial 2Center for Emerging and Re-emerging Efficacy of Novel Lactate Dehydrogenase Infectious Diseases, Division of Allergy Inhibitors and Infectious Diseases, Department of Medicine, University of Washington, Seattle, William Witola1, Mr. Kun Li1, Dr. Sara Nader1, United States, 3Institute of Parasitology, Ms. Xuejin Zhang1, Dr. Chi Yong Kim1 Vetsuisse Faculty, University of Berne, Berne, 1University Of Illinois At Urbana-champaign, Switzerland College of Veterinary Medicine, Champaign, United States The chemo-prophylactic control of cystoisosporosis in the European Union Cryptosporidium parvum is a highly prevalent currently relies on the use of toltrazuril. zoonotic and anthroponotic protozoan Recently, resistance against toltrazuril in a parasite that causes a diarrheal syndrome in Dutch field isolate of Cystoisospora suis has children and neonatal livestock, culminating been described, and therefore, the need of new in growth retardation and mortalities. Despite therapeutics to overcome the limited treatment the high prevalence of C. parvum, there are options is of immediate importance. Bumped no fully effective and safe drugs for treating kinase inhibitors (BKIs) specific for parasite infections, and there is no vaccine. We have calcium-dependent protein kinases (CDPKs) previously reported that the bacterial-like have already been shown to reduce infection in C. parvum lactate dehydrogenase (CpLDH) several apicomplexan parasites of human and enzyme is essential for survival, virulence and veterinary importance, including Toxoplasma growth of C. parvum in vitro and in vivo. In gondii, Neospora caninum, Cryptosporidium the present study, we screened compound parvum, and Sarcocystis neurona. In the present libraries and identified inhibitors against the study, the efficacy, safety and pharmacokinetics enzymatic activity of recombinant CpLDH of BKI 1369 in piglets experimentally infected protein in vitro. We tested the inhibitors for with Cystoisospora suis was determined using anti-Cryptosporidium efficacy using in vitro an established animal infection model, and in infection assays of HCT-8 cell monolayers cell culture. Five-day treatment with BKI 1369 and identified compounds A1.0 and C2.0 that effectively suppressed oocyst excretion and inhibited the proliferation of intracellular C. diarrhea and improved body weight gains in parvum in vitro, with IC50 values of 14.88 treated piglets without obvious side effects and 72.65 M, respectively. At doses tolerable for both toltrazuril-sensitive (Wien-I) and in mice, we found that both A1.0 and C2.0 resistant (Holland-I) C. suis strains. In vitro, consistentlyµ reduced the shedding of C. BKI 1369 inhibited merozoite proliferation in parvum oocysts to undetectable levels in intestinal porcine epithelial 1 (IPEC-1) cells by infected immunocompromised mice’s feces, at least 50% at a concentration of 40 nM and and prevented intestinal villous atrophy as

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 25 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

well as mucosal erosion due to C. parvum. and amplification of the C. suis gene Together, our findings have unveiled promising CSUI_005805. Three different standards – anti-Cryptosporidium drug candidates that can DNA from merozoite dilutions, direct DNA be explored further for the development of the dilution standards and a cloned plasmid much needed novel therapeutic agents against standard – were compared for efficacy and C. parvum infections. sensitivity. On the basis of this assay, in vivo efficacy testing can be tailored to reduce the number of animals used and provide OA04.03 A Cystoisospora Suis In Vitro Assay information on the possible mechanisms of for Compound Screening: The Example of action of tested compounds. Bumped Kinase Inhibitor BKI 1369

Mrs. Baerbel Ruttkowski1, PhD Aruna OA04.04 New Treatment Options for Shrestha1, Prof Anja Joachim1 Piglet Coccidiosis: Injectable Toltrazuril in 1Institute of Parasitology, Department of Combination With Iron (As Gleptoferron) Is Pathobiology, Vetmeduni Vienna, A-1210 Effective Against Experimental Cystoisospora Wien, Austria Suis Infections on the 1st or 3rd Day of Life

Currently the only drug registered for Prof. Anja Joachim1, Dr. Nicolas Guerra2, treatment of piglet coccidiosis caused by Dr. Barbara Hinney1, Dr. Adnan Hodžić1, Dr. Cystoisospora suis is toltrazuril. However, Hamadi Karembe2, Dr. Aruna Shrestha1, Dr. in the light of the first recorded toltrazuril- Daniel Sperling2 resistant field isolates alternatives must 1Institute of Parasitology, Department of urgently be sought. Screening for compounds Pathobiology, Vetmeduni Vienna, A-1210 in vitro can guide the selection of compounds Wien, Austria, 2Ceva Santé Animale, 33500 with assumed efficacy in vivo and give Libourne, France hints at mechanisms of efficacy and stage- specific activity. A previously developed in Piglet coccidiosis is one of the most common vitro cultivation system for C. suis, using diarrheal diseases of suckling piglets. sporozoites obtained from oocysts ex vivo to Infection causes poor growth and seriously infect semi-permanent IPEC-J1 cells to sustain compromises animal health. Where available, the development of all endogenous life-cycle toltrazuril is applied orally in a metaphylactic stages, was adapted to quantify merozoites oral single-use individual application to developed from initial sporozoite infection. prevent disease. Recently, an injectable A serial dilution of sporozoites was used to toltrazuril has been developed for parenteral adapt the culture conditions and to evaluate application on the 1st - 3rd day of life (dol) the growth curve of merozoites during 5-9 together with iron for anemia prevention in a days of cultivation. Along with in vivo efficacy single application (Forceris®, 45 mg toltrazuril testing, the bumped kinase inhibitor BKI-1369 + 200 mg iron/piglet). In two experimental was evaluated in vitro for efficacy against trials with infection of piglets on the 1st resp. C. suis. The standard treatment for five days 3rd dol (1000 C. suis oocysts/piglet) the from the day of infection could be reduced efficacy of this product (application on the in vitro to three days, starting two days post 2nd dol) was evaluated in comparison to a infection in vitro. Compound testing includes control group (iron treatment only). Oocyst cell toxicity of the compound and its solvent, excretion, diarrhea and weight gain were titration of effective drugs with determination compared between groups. The presence of of IC 50/IC90 and the evaluation of bacterial enteropathogens was determined treatment duration. Test readout was either in selected samples. For each group, 10-13 microscopical evaluation by counting of free piglets finished the study and were included merozoites harvested 9 days after infection in the evaluation. In the control groups, all from culture supernatants or by quantitative piglets excreted oocysts and had diarrhea real-time PCR with determination of the for at least one day. In the treated groups total amount of merozoites in supernatant oocyst excretion was completely suppressed and cells followed by DNA preparation and diarrhea was reduced to a one resp. two

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 26 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

piglets and five resp. three diarrheic samples. T. gondii exosomes were recognized by the Differences between treated and control antisera against rTgSRSs protein. Indirect groups were significant for all parameters. immunofluorescence assay results indicated Weight gain was suppressed during the acute that three TgSRSs were located either on cell phase of infection (8th – 15th dol), resulting surface or cytoplasm of T. gondii tachyzoites. in reduced body weight in the controls Applying CRISPR/CAS9 technology, three compared to the treated groups by the end gene deletion and overexpression plasmids of the study (29th dol). Infection on the 1st targeting TgSRSs were constructed. To dol induced more intense oocyst excretion understand the complex mechanisms while diarrhea was less severe, probably due behind the pathogenicity of T. gondii, and to insignificant levels of Cl. perfringens type A thereby facilitate the development of proper and E. coli compared to the trial with infection preventive measures against Toxoplasmosis, on the 3rd dol. Forceris® treatment on the 2nd we analyzed the structural characteristics dol was highly effective against experimental and functions of these proteins in toxoplasma C. suis infections on the 1st – 3rd dol in the invasion, reproduction and interaction with presence of bacterial enteropathogens. host cell proteins, in addition to other possible roles of these three rTgSRSs.

OA04.05 Molecular Mechanisms Involved in Interaction of SRSs of Toxoplasma Gondii OA04.06 TKL1-Deficient Toxoplasma Gondii Exosomes with Host Proteins Is a Potential Live-Attenuated Vaccine Against Acute, Chronic and Oocysts-Caused Houshuang Zhang1, Xiaoxing Lin1, Qingcan Congenital Toxoplasmosis in a Murine Model Wang1, Jie Cao1, Jinlin Zhou1 1Shanghai Veterinary Research Institute, Dr. Jin-lei Wang1, Mr. Qin-Li Liang1, Miss Ting- Chinese Academy Of Agricultural Sciences, Ting Li1, Dr. Jun-Jun He1, Miss Meng-Jie Bai1, Shanghai, China Prof. Hany M. Elsheikha2, Prof. Xing-Quan Zhu1 1State Key Laboratory of Veterinary Etiological Knowledge of the exosomes secreted by T. Biology, Key Laboratory of Veterinary gondii parasites or T. gondii-infected host Parasitology of Gansu Province, Lanzhou cells is still very limited. Apart from their Veterinary Research Institute, Chinese Academy role in cell-to-cell communication, exosomes of Agricultural Sciences, Lanzhou, China, secreted by T. gondii are able to stimulate 2Faculty of Medicine and Health Sciences, host immune response directly or indirectly, School of Veterinary Medicine and Science, via dendritic cells and other APCs, which University of Nottingham, Sutton Bonington could also lead to specific pathological Campus, Loughborough, United Kingdom changes in the host. In a previous study, we successfully identified some surface Toxoplasma gondii is an important pathogen antigen 1-related sequences (SRSs) proteins for which a vaccine would address a in T. gondii exosomes, beside other T. gondii significant unmet public health and medical specific proteins. T. gondii SRS proteins are need. We tested whether deletion of tyrosine involved in the process of attachment to kinase-like 1 (tkl1) can be an effective strategy host cells, along with stimulating immune for developing a live-attenuated vaccine, reactions and regulating parasite virulence, with controllable attenuation and potent which is thought to promote the formation immunogenicity. The protective efficacies of of tissue cysts in intermediate hosts in order vaccination using tachyzoite against to establish persistent, latent infections that acute, chronic and congenital toxoplasmosis facilitate transmission of infection to the in Kunming mice wereRHΔtk1 evaluated. Mice definitive host. In our current study, three vaccinated with developed a strong SRSs were identified and characterized humoral and cellular response as indicated by from T. gondii (designated TgSRSs). a high level of T. RHΔtk1gondii-specific IgG, IL-2, IL-12, Recombinant TgSRSs (rTgSRSs) were IFN- and IL-10. All vaccinated mice survived expressed in Escherichia coli and purified. a lethal challenge with 103 tachyzoites of RH Three SRSs native proteins of T. gondii or or ToxoDB#9γ (PYS or TgC7) strains, as well

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 27 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

as 100 tissue cysts or oocysts of Pru strain, California counties. To better understand the whereas all non-vaccinated mice infected diagnostic value of HW screening results in low- with these parasites died. Vaccination also prevalence areas, in-clinic SNAP®4Dx®Plus-HW protected against cysts or oocysts caused results from 2013 to 2016 were examined for chronic infection, reduced the vertical California and for Scandinavia where virtually transmission caused by oocysts, increased the no autochthonous HW transmission is believed litter size, and maintained pups’ body weight to occur. For this analysis, all positive HW tests in dams challenged with 10 oocysts on day 5 from Scandinavia were conservatively treated of gestation. In contrast, all non-vaccinated as false positives. The observed specificity + oocysts-infected dams had abortions and was defined as the proportion of Scandinavia no fetus survived. Vaccinated dams remained HW tests that returned a negative result. healthy after infection and their brain cyst Using the observed Scandinavian specificity, burden was significantly reduced compared SNAP®4Dx®Plus-HW results from California with with non-vaccinated + oocysts-infected dams. the stated test sensitivity, we calculated the A single vaccination of mice with RH tkl1 positive predictive values in California counties. tachyzoites induced a balanced and strong Of the 14,778 Scandinavian tests results, 18 were immunity against T. gondii with differentΔ positive giving a positive rate of 0.12% and an stages, and protected mice against congenital observed specificity of 99.88%. For the same oocysts infections. This is, to the best of period, there were 301,997 test results from our knowledge, the first to demonstrate the 476 California veterinary clinics with 1,742 being efficacy of live-attenuated vaccine against HW positive with a positive rate of 0.58%. The oocysts-caused congenital toxoplasmosis. positive rate for the SNAP®4Dx®Plus-HW tests in California was significantly higher than that observed in Scandinavia by chi-square test (P < OA05 Worldwide Vector-Borne 0.001). Therefore, the higher than expected rate Infections in Companion Animals of SNAP®4Dx®Plus-HW test positives cannot be solely attributed to a lack of specificity (false July 8, 2019, 11:00 - 12:30 positives) but strongly suggests the actual Breakout Room 5, Meeting Rooms KLOP, Level 4 presence of Dirofilaria immitis in California. The results support that any HW-positive test results on the SNAP®4Dx®Plus-HW, due to its OA05.01 Highly Specific Screening Tests high specificity, are highly probable to be true Maintain Actionable Positive Predictive Values positives. in Areas with Low Heartworm Prevalence

1, Jennifer Braff2, Prof, Dwight D. Bowman OA05.02 Trends in Global Clinic-Based Jesse Buch2, Andrei Rakitin2, Ramaswamy ELISA Seropositive Rates for Canine Vector- Chandrashekar2, Melissa Beall2 Borne Disease 1Cornell University, ITHACA, United States, 2IDEXX Laboratories, Inc, Westbrook, United Dr. Jesse Buch1 States 1IDEXX Laboratories, Inc., Westbrook, United States At low prevalence, the positive predictive value of a diagnostic screening test is directly Current trends in animal relocation and proportional with disease prevalence within vector range expansion have increased a given region. Consequently, where the the importance of studying the global prevalence of heartworm infection has dropped epidemiology for canine vector-borne disease below the claimed false positive rate (1 – (CVBD). In some regions, epidemiological specificity) for a screening test, rare positive data on vector-borne zoonotic pathogens test results may be suspected as false positives. is available to a far greater extent for canine This study re-evaluated the field-observed populations than humans. Thus, CVBD data is specificity of the SNAP® 4Dx®Plus test for becoming an important component of public heartworm (HW) antigen and estimated the health as an increasing emphasis is being local positive predictive value (PPV) for specific placed on studying canine populations as

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 28 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

sentinels for human vector-borne disease. This and roundworm 9.1%. Larva were observed in talk will review recent global trends in CVBD 3 of 49 fecal samples assessed by Baermann clinic-based ELISA results for Dirofilaria immitis technique for Aerulostrongylus abstrusus but antigen and antibodies to Borrelia burgdorferi, none were identified as A. abstrusus. Results Ehrlichia spp., Anaplasma spp. and Leishmania. from IDEXX RealPCRTM analysis of 69 blood The trends are based on more than 60 million samples yielded: Mycoplasma haemominutum in-clinic serological CVBD test results reported 20.3%, Bartonella sp. 17.4%, Mycoplasma from more than 11,000 veterinary clinics haemofelis 7.2%, feline panleukopenia 4.4%, located in 79 countries from 7 regions. The Mycoplasma turicensis 4.3%, FeLV 1.4% and strengths and limitations of these data will also Ehrlichia ewingii 1.4%. Forty-two (60.9%) be discussed. cats were positive for ≥1 infectious organism. Of these, 18 were co-infected with up to 4 organisms. Feral cats were infected with OA05.03 Comprehensive Survey of parasites and infectious organisms more Parasitic and Infectious Diseases in Feral & frequently than domesticated cats. This is the Domesticated Cats in Grenada, West Indies most comprehensive assessment of infectious diseases in Grenadian felids. The results of Dr. Tara Paterson1, Tamara Hockley1, Dr. Dave this investigation will aid in the diagnosis and Elsemore2, Dr. Jancy Hanscom2, Dr. Phyllis treatment of cats in Grenada and the broader Tyrrell2, Dr. Christian Leutenegger2, Dr. Caribbean region. Ramaswamy Chandrashekar2 1St. George’s University, School Of Veterinary Medicine, St. George’s, Grenada, 2IDEXX OA05.04 Prevalence of Babesia Spp. And Laboratories, Inc., Westbrook, United States Clinical Characteristics of Babesia Microti- Like Infections in North American Dogs Little data exists on feline infectious diseases in the Caribbean even though some are Dr. Nanelle R. Barash1, Dr. Barbara Qurollo1, Ms. zoonotic and are of significant public health Brittany Thomas1, Dr. Adam J. Birkenheuer1, Dr. concern. The objective of this study was Edward B. Breitschwerdt1, Ms. Erica Lemler1 to survey domesticated and feral cats for 1North Carolina State University, College of parasitic and infectious diseases using Veterinary Medicine, Department of Clinical a comprehensive diagnostic approach. Sciences, Raleigh, United States Seventy-one cats (46 feral, 25 domestic) presenting to the St. George’s University Babesia microti-like infection, reported in (SGU) Small Animal Clinic for care or European dogs and North American foxes, sterilization through the SGU Feral Cat Project has rarely been reported in domestic North were sampled. Sixty-five blood samples were American dogs. We determine the prevalence tested using the IDEXX SNAP® Feline Triple® of Babesia spp., including B. microti-like, in test – 13.8%, 13.8% and 9.2% were positive a large convenience set of North American for Dirofilaria immitis, feline leukemia virus dog blood samples submitted to a reference (FeLV) and feline immunodeficiency virus, laboratory for vector-borne disease testing respectively. Fifty-seven fecal samples were between June 2015 and June 2018. In addition evaluated at SGU using centrifuged fecal to Babesia spp. testing, comprehensive floatation - 82.5% of cats were infected canine vector-borne disease diagnostics were with ≥1 parasite. The most common parasite performed to identify co-infecting pathogens. observed was hookworm (Ancylostoma sp., Babesia spp. DNA was PCR amplified from 78.9%), followed by Mammomonogamus sp. 269/9376 (2.9%) dogs. The most prevalent (40.4%), whipworm (Trichuris sp., 33.3%), Babesia species were B. gibsoni (187; 2.0%) roundworms (Toxocara cati 8.8%, Toxascaris and B. microti-like (48; 0.51%), of which leonina 1.8%), Isospora felis (8.8%), and 30 (0.32%) dogs were co-infected both Dipylidium caninum (3.5%). Fecal Dx® antigen Babesia spp. Limited clinicopathologic data testing was performed on 44 fecal samples in B. microti-like infected dogs revealed at IDEXX Laboratories with the following proteinuria (75%), hyperglobulinemia (67%), results: hookworm 56.8%, whipworm 29.5%, thrombocytopenia (60%), anemia (58%), and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 29 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

hypoalbuminemia (56%). Co-infections with province (OR 4.93 vs. Avellino, 95% CI 3.18- Mycoplasma, Dirofilaria immitis, or Wolbachia 7.64), long coat length (OR 1.79 vs. short, and co-seroreactivity to Bartonella, Ehrlichia, 95% CI 1.16-2.77), seropositivity for E. canis and Rickettsia spp. were documented in B. (OR 2.84, 95% CI 1.72-4.69), and hound breed microti-like infected dogs. In North America, (OR 2.97 vs. pointing, 95% CI 1.88-4.70). dogs can be infected with B. microti-like and The significant association with E. canis develop clinicopathological abnormalities seropositivity and lack of association with consistent with babesiosis caused by other the observed presence of ticks could be an Babesia spp. indication that the duration of exposure to the vector is important. Although hunting dogs in Southern Italy are frequently exposed to OA05.05 Hepatozoon Canis in Hunting Dogs H. canis, infections could be underestimated From Southern Italy: Distribution and Risk because clinical signs overlap with other tick- Factors borne diseases. Further studies are necessary to determine the epidemiological relationships Dr. Laura Pacifico1, Dr. Melissa Beall2, Dr. between hunting dogs and wild animal Francesco Buono1, Dr. Jennifer Braff2, populations sharing the same environments in Giovanni Sgroi1, Dr. Jesse Buch2, Dr. Diego Southern Italy. Piantedosi1, Dr. Benedetto Neola3, Dr. Christian Leutenegger2, Dr. Ramaswamy Chandrashekar2, Dr. Vincenzo Veneziano1 OA06 Canine Heartworm II 1University of Naples Federico II, Napoli, Italy, 2IDEXX Laboratories, Inc., Westbrook, ME July 8, 2019, 13:30 - 15:30 04092, United States, 3Istituto Zooprofilattico Plenary Hall, Madison Ballroom (ABCD), Level 4 Sperimentale del Mezzogiorno, Portici, Italy

Hepatozoon canis infects domestic and wild OA06.01 Colorado Dog Importation and carnivores in Europe. Hepatozoon spp. are Increases in Heartworm Prevalence mainly transmitted through ingestion of infected ticks containing mature oocysts. Jason Drake1, Rudolph Parrish1 In Europe, the principal vector of H. canis is 1Elanco Animal Health, Greenfield, United Rhipicephalus sanguineus, and the distribution States of the disease correlates with the range of the vector. However, H. canis has been reported Animal welfare groups, attempting to increase outside of these areas, often related to adoptions and reduce euthanasia, transport infection in wild carnivores. Because hunting thousands of rescued dogs state-to-state dogs can act as sentinels for monitoring annually. A recent survey reported that wildlife diseases, this study was performed approximately only 1/3 of animal welfare to determine the prevalence and risk factors groups test, treat or provide prevention for associated with H. canis infection in hunting heartworm (Dirofilaria immitis) infection dogs from Southern Italy. Blood was collected prior to shipping dogs. Studies have reported from 1,433 dogs in Napoli, Avellino and prevalence of heartworms in dogs in shelters Salerno provinces as part of a Campania or rescued following natural disasters ranging region hunting dog health assistance program. from 14.6 – 48.8%. For this study, we obtained Samples were tested by real-time polymerase data from Companion Animal Parasite chain reaction (PCR) to amplify H. canis Council (CAPC) heartworm prevalence DNA. PCR identified 200 positives, indicating maps and from the Colorado Department a prevalence of 14.0%. Multiple logistic of Agriculture Pet Animal Care Facilities regression on H. canis PCR results determined Program (PACFA) and investigated changes that province (p<.0001), coat length (p=.026), in heartworm prevalence and the number E. canis serological status (p<.0001), and of dogs shipped into Colorado. The CAPC breed (p<.0001) were significant predictors heartworm maps reveal that the prevalence of of H. canis infection. Risk factors for positive heartworms in Colorado dogs has increased H. canis PCR results included living in Salerno 67.5% from 2013-2017. The PACFA data from

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 30 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

2014-2017 show that over 130 animal welfare overcome these diagnostic shortcomings, organizations in Colorado received more multiple real-time PCR-based diagnostic than 114,000 dogs from out-of-state sources, assays have been developed in recent years. representing approximately 9.5% of the Capable of detecting D. immitis in both total estimated 2017 Colorado population of vector mosquitoes and definitive hosts, these nearly 1.2 million dogs. Three large Colorado- assays provide a powerful new diagnostic based organizations responded to requests option. However, to date, all such diagnostics for details regarding the originating states have utilized sub-optimal mitochondrial from which they received dogs. The majority or ribosomal targets. Maximizing the of these dogs were apparently shipped to diagnostic sensitivity and specificity of real- Colorado from states with higher heartworm time PCR requires the targeting of highly prevalence. New Mexico represented the divergent, high copy-number repetitive DNA source of the greatest number of relocated sequences. Accordingly, we now describe dogs, accounting for just over 30%. Nearly the development of a novel real-time PCR half (49%) of the dogs relocated by these diagnostic, targeting a highly repetitive three organizations came from either Texas genomic DNA element. This element or Oklahoma. Animal welfare organizations was identified using a next-generation and veterinarians should increase testing and sequencing-based approach that we have prevention of heartworm infections in dogs previously employed to improve diagnostic prior to, and following, shipment from areas options for human-infecting soil-transmitted with endemic heartworm. Repeated testing helminthes. Our target represents the most is recommended due to the 6-month pre- highly repetitive DNA element within the patent-period associated with D. immitis. D. immitis genome. Accordingly, assay Veterinarians and pet owners should increase sensitivity and specificity has been maximized vigilance with heartworm testing and targeting this element. Comparative testing prevention, even in areas with historically low against genomic DNA isolated from various heartworm prevalence. Movement of dogs filarial nematodes confirmed assay specificity from highly endemic areas may increase the and a panel of field-collected samples risks of local transmission. was employed to demonstrate improved sensitivity of detection vs. established molecular diagnostic options. Furthermore, OA06.02 Improved Real-Time PCR due to the evolutionarily divergent nature Diagnostics for Dirofilaria Immitis Utilizing a of such DNA targets, the adaptability of this Next-Generation Sequencing-Based Approach assay to conventional PCR-based approaches to Target Selection and Assay Design was also evaluated. Such adaptability could expand the utility of this assay to settings Mr. Nils Pilotte1, Dr. Lori Saunders2, Dr. Steven where real-time PCR is not available. Williams2 1University Of Massachusetts-Amherst, Amherst, United States, 2Smith College, OA06.03 Do Canids Infected with Dirofilaria Northampton, United States Immitis Release Unique Volatile Organic Compounds in Their Breath? Dirofilaria immits is a mosquito-borne pathogen with a wide range of hosts in Dr Sarah Zohdy1,2, Melissa Boersma3, Dr. both the Canidae and Felidae. Current Heather Stockdale Walden4, Dr Lindsay diagnostic practices involve either direct Starkey2, Professor Byron Blagburn2 microscopic observation of microfilaria or 1School of Forestry and Wildlife Sciences, the use of ELISA assays targeting antigens Auburn University, Auburn, United States, released by adult female worms. While 2Department of Pathology, College of commonly employed, the diagnostic use Veterinary Medicine, Auburn University, of microscopy can lead to misdiagnosis or Auburn, United States, 3Department of under diagnosis, and significant concerns Chemistry and Biochemistry, College of regarding the sensitivity and specificity of Science and Mathematics, Auburn University, available ELISA tests exist. Attempting to Auburn, United States, 4Department of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 31 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Comparative, Diagnostic & Population OA06.04 Collection and Molecular Detection Medicine, College of Veterinary Medicine, of Dog Heartworm, Dirofilaria immitis, DNA University of Florida, Gainesville, United States in Wild Caught Mosquitoes

Dirofilaria immitis, causative agent of Phillip Kaufman1, Chris Holderman2, canine heartworm disease, is a mosquito Nicole Abruzzo1, Noor Abdelsamad1, Peter transmitted parasite. Mosquito vectors use DiGennaro1 host semiochemical signals such as CO 1University Of Florida, Gainesville, United and odorant compounds to locate viable States, 2Central Life Sciences, Dallas, United hosts. Here, we aim to examine the role₂ of States canine heartworm infection on the chemical fingerprints of breath volatiles which may Dirofilaria immitis, the causative agent of facilitate mosquito attraction and therefore dog heartworm disease, is responsible for a parasite transmission dynamics. Specifically, disproportionate amount of canine morbidity we hypothesize that infected dogs will and mortality; in addition to being expensive release unique volatile organic compounds to treat, severe infections are often fatal. (VOCs) in their breath which are mosquito Much is known about the pathogen in the attractants and may be used to non-invasively canine host, yet little is known on the basic detect heartworm infection status. We biology of the nematode in the mosquito developed a rapid (under 2 minutes) method vector. Thus, to evaluate the effectiveness of to capture and store 1 liter of exhalant from collection techniques on ability to capture dogs for downstream thermal desorption dog heartworm infected mosquitoes, and processing by gas chromatography mass we conducted a field study spanning 111 spectrometry (GC-MS). Using this method weeks. Four methods were used to collect we sampled 25 dogs with known heartworm mosquitoes; two aspirators types, sweep infection status, and identified the compounds netting, and a CDC trap. Sites with canines using automated and manual extraction present in either residential yards (n=4) or methods (AMDIS). Over 500 unique VOCs dog kennel facilities (n=3) were utilized. were identified and classified into chemical Collected mosquitoes were sorted by site, subclass. GC arrays (heat maps) of the trap, species, and date, then pooled into volatile breathomes were generated and show groups of up to 25 individuals. Mosquito head significantly more unique VOCs in infected and thorax pools were extracted for DNA, dog breath than in non-infected dogs, and that was screened using currently available more phenols, which are known attractants. protocols. These protocols were found However, contrary to our hypothesis, alkane unreliable, thus, we developed a novel qPCR hydrocarbons such as decanes and nonanes, primer and probe set. known mosquito repellant compounds, were found in non-infected dogs and Using this method, the original samples were these compounds were largely absent re-assayed and provided 644 positive pools. from infected dogs. The emerging field of Approximately 10% of positive samples were breathomics has been used to examine VOCs confirmed by Sanger sequencing. Twenty-three as biomarkers of diseases in humans; however, species tested positive for dog heartworm DNA, to our knowledge no one has used breath including a new association with Wyeomeyia biomarkers as signals of pathogen detection mitchellii. Aedes atlanticus, Anopheles crucians, in animals. Here, we find that breath VOC and Culisieta melanura composed nearly 28% composition differs significantly between D. of the total collection, yet were responsible for immitis infected and non-infected dogs as 66% of the qPCR positive pools. Infection rates does the presence of mosquito attractant for commonly collected mosquitoes ranged and repellant compounds. These results up to 2.4% with more rarely collected species suggest that mosquito-host interactions ranging up to 20%. may be manipulated by the metabolic by- products of parasitic infection driving parasite We present a new qPCR primer and probe transmission dynamics. set for detecting dog heartworm DNA. The CDC trap appears to be the most effective

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 32 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

collection method, yet other techniques OA06.06 Canine Heartworm Infection and should be utilized when targeting mosquito Prophylaxis Use among Pet Caretakers from species that this trap is biased against. the Cumberland Gap Region of Tennessee, USA

OA06.05 Evaluation of Heat Treatment Briana Cappiello1, Ariel Hudson1, Dawn of Serum Prior to Testing for Detection of Spangler1, Karen Gruszynski1, Charles Faulkner1 Antigen of Dirofilaria immitis in Necropsy 1Lincoln Memorial Univ College Vet Medicine, Positive and Negative Dogs from Florida, USA. Harrogate, United States

Mr. Jeff Gruntmeir1, Dr. Maureen Long1, Dr. The incidence of canine heartworm (CHW), Byron Blagburn2, Dr. Heather Walden1 Dirofilaria immitis, infection continues to 1University of Florida, College of Veterinary increase in the United States, despite the Medicine, Gainesville, United States, 2Auburn availability of effective prophylaxis. Current University, College of Veterinary Medicine, reports of CHW prevalence based on use Auburn, United States of veterinary services likely underrepresent the true magnitude of the issue because pet Multiple studies have used heat treatment to dogs not receiving regular veterinary care, determine changes in prevalence of Dirofilaria and dogs in residing in animal shelters are immitis antigen in serum of naturally infected often excluded from distribution maps. This dogs in the absence of necropsy data. In order study estimated the prevalence of CHW to determine changes in test sensitivity and in a pet dog population and prophylactic specificity following heat treatment, serum use by pet caretakers, irrespective of their from 547 dogs (259 necropsy positive and 288 use of veterinary services. Pet dogs from necropsy negative samples) were evaluated the Cumberland Gap region in Tennessee, using the DiroCHEK® (Zoetis) antigen assay. USA were serologically tested for CHW and Positive results were determined visually their caretakers interviewed about monthly and by spectrophotometry at 650nm at 5 CHW prophylaxis use and perceptions minutes. All samples were prepared and regarding knowledge and prevention of subjected to heat treatment by an operator CHW. This was accomplished by pedestrian that was unaware of necropsy results using neighborhood survey. CHW prevalence was 300 l of serum. Positive antigen results were 2% (3/125) in the tested population. Non-use interpreted based on necropsy results for of monthly prophylaxis was present in 42% collectionμ years 2011-2015 (n=223) and 2017- of surveyed pet caretakers, implying that 2019 (n=36). Necropsy negative samples from 54 of the 125 dogs tested are potentially 2011-2015 (n=224) were randomly selected at risk for infection with CHW. Significant from 367 available samples determined as (p < 0.05) predictors of prophylaxis use antigen negative. Of the necropsy positive identified by a logistic regression model samples, 225/259 (86.9%) and 245/259 include household income, use of veterinary (94.6%) samples tested positive before and services, altered reproductive status (spay/ after heat treatment, respectively. For the neuter), knowledge of CHW, and confinement necropsy negative samples, 288/288 (100%) outdoors. Pet caretakers utilizing regular and 277/288 (96.2%) tested negative before veterinary services were more likely to and after heat treatment, respectively. Using perceive CHW as an important health factor, necropsy status as the reference, sensitivity and administer monthly prophylaxis to their was improved by 7.7% and specificity pets. These results underscore the importance decreased by 3.8%. Heartworm intensities of veterinary service mediated interaction ranged from 1-30 for post-heat antigen with pet caretakers to enhance awareness positive samples and 1-62 for those remaining of CHW and facilitate adoption of effective antigen negative. Results will be discussed prophylactic programs. in context of parasite life cycle, necropsy results, and application of optical density as an indicator of positive or negative infection.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 33 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA06.07 First Case of a Natural Non Patent infections were clinically relevant. As A. Infection in a Domestic Cat (Felis catus) abstrusus and A. vasorum are both gastropod With the Canid Heart Worm Angiostrongylus transmitted nematodes, they may share the Vasorum same intermediate hosts within overlapping areas. In addition, especially chronic A. Dr. med. vet. Emily Gueldner1, Dr. med. vet. abstrusus infected cats become non-patent Carole Schuppisser2, Prof. Nicole Borel3, and do not excrete L1. Considering that patent Dr. med. vet. Monika Hilbe3, Prof. Manuela A. vasorum infections are widespread in the Schnyder1 dog and fox population in Switzerland but 1Institute of Parasitology, Vetsuisse Faculty, are apparently not patent in cats, we cannot University Of Zurich, Zürich, Switzerland, exclude that infections with A. vasorum may be 2Clinic for Small Animal Internal Medicine, misinterpreted and occur more frequently than Vetsuisse Faculty, University of Zurich, expected, particularly in highly endemic areas. Zürich, Switzerland, 3Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Zürich, Switzerland OA06.08 Heartworm Control in Grenada, West Indies: Results of a Field Study Cardiopulmonary nematodes in cats include using Imidacloprid 10% + Moxidectin 2.5% different parasite species affecting feline (Advantage Multi®, Advocate®) Spot-on and lungs and the heart, with the metastrongyloid Doxycycline for Naturally Acquired Dirofilaria Aelurostrongylus abstrusus being the most immitis Infections frequent feline lungworm worldwide. The present case report describes a 11 month old Dr. Tara Paterson1, Dr. Christina Fernandez1, male neutered European short hair cat which Dr. Peter Burnett1, Dr. Lindyann Lessey1, Dr. presented with generalized subcutaneous Tamara Hockley1, Dr. Roland Schaper2 oedema and pleural and peritoneal effusions. 1St. George’s University, School of Veterinary According to clinical examination, abdominal Medicine, St. George’s, Grenada, W.I., 2Bayer imaging and laboratory analyses, a tentative Animal Health GmbH, Leverkusen, Germany diagnosis of severe glomerulopathy with massive proteinuria was made. Due to Melarsomine dihydrochloride (Immiticide®) worsening of the clinical signs despite is an effective adulticidal therapy in dogs therapeutic interventions and a poor with canine heartworm disease (CHD), prognosis, the cat was euthanized. Necropsy however the risk of post-therapy pulmonary and histological examinations revealed severe thromboembolism and expense of treatment bilateral collagenofibrotic glomerulopathy, warrants exploration of alternative treatment generalized oedema and a focal verminous options for CHD. This randomized, controlled pneumonia with thrombosis in arterial lung field study compares the adulticidal efficacy vessels containing nematode cross sections. of 10% imidacloprid + 2.5% moxidectin A serum sample was tested for the presence (Advantage Multi®, Advocate®) and oral of antibodies against the cat lungworm A. doxycycline with that of Immiticide®. Thirty abstrusus, resulting negative. Genetic analyses naturally-infected dogs with class 1, 2 or confirmed the presence of nematode DNA; early class 3 CHD were evaluated for up to and, after exclusion of common lung and 18 months. Dogs were randomly assigned heart parasites occurring in cats, DNA of the to a control (CP, n=15) or investigational canid heart worm nematode A. vasorum was treatment (IVP, n=15) group. CP dogs received identified. This is the first description of a two injections of Immiticide® (2.5mg/kg) naturally occurring infection with A. vasorum 24-hours apart –according to US label- in a cat. and monthly ivermectin/pyrantel pamoate (Heartgard® Plus). IVP dogs were treated with Previous experimental studies demonstrated oral doxycycline (10mg/kg q12h for 28 days) the development of adult male and female A. and topical Advantage Multi® / Advocate® vasorum worms containing eggs in cats, but once monthly for 9 months. Dogs were no larval excretion in the faeces. Although evaluated up to 18 months - monthly for the cats did not become patent, A. vasorum first 9 months, then every 3 months. IVP

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 34 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

data was analyzed for non-inferiority defined had lower blood eosinophil counts and as ≤15% margin of difference. Parasiticidal higher mean worm burdens than control efficacy was based on antigen status using sheep vaccinated with Quil-A adjuvant alone. the IDEXX PetChek® 34 Heartworm-PF In addition, adult worms in IL-5 vaccinated Antigen test. One dog in each treatment sheep were significantly longer with higher group remained antigenemic by month 15. eggs in utero in female worms. These results Based on antigen status, non-inferiority was suggest that eosinophil plays a direct role in confirmed at months 12, 15 and 18. Non- effective control in H. contortus infection in inferiority was also demonstrated at months CBH sheep. 12 & 18 based on results of heat-treatment of antigen-negative samples. Non-inferiority of the ultrasonography data at month 12 OA07.02 Fasciola Hepatica Infection in was demonstrated as no parasites were Cattle: Analysing Responses of Peripheral observed in 50% and 60% of IVP and CP Blood Mononuclear Cells (PBMC) Using a dogs, respectively. Monthly application Transcriptomics Approach of Advantage Multi®/Advocate® combined with oral doxycycline has proven to be an Dr. Andres Garcia-Campos1, Dr. Carolina effective, safe and affordable therapeutic Correia2, Ms. Amalia Naranjo-Lucena1, Dr. alternative for the treatment of CHD and Laura Garza-Cuartero1, Dr. Gabriella Farries2, should be considered particularly in cases John Browne2, Professor David MacHugh2,3, where finances or debilitated health preclude Professor Grace Mulcahy1,3 treatment of an infected individual. This study 1UCD School of Veterinary Medicine, University was supported by Bayer Animal Health. College Dublin, Belfield, Dublin, Ireland,2 UCD School of Agriculture and Food Science, University College Dublin, Belfield, Dublin, OA07 Host Responses Against Ireland, 3UCD Conway Institute of Biomolecular Helminths in Ruminants and Biomedical Research, University College Dublin, Belfield, Dublin, Ireland July 8, 2019, 13:30 - 15:30 Breakout Room 2, Hall of Ideas E&H, Level 4 The parasitic helminth Fasciola hepatica (liver fluke) causes economic loss to the livestock industry globally and also represents OA07.01 IL-5 Dependent Modulation a zoonotic disease. New control strategies of Eosinophil Responses Augments such as vaccines are urgently needed, due Haemonchus Contortus Infection in Sheep to the rise of drug resistance in parasite populations. Vaccine development requires Dr Jorge Francisco González1, Dr Julia a comprehensive understanding of the Natividad Hernández1, Dr Els Meeusen2, Dr. immunological events during infection. David Michael Piedrafita2 Previous in vivo studies by our group have 1Universidad De Las Palmas De Gran Canaria, investigated global differentially expressed Arucas, Spain, 2Federation University, genes (DEGs) in ovine peripheral blood Churchill, Australia mononuclear cells (PBMC) in response to both acute and chronic F. hepatica infection. This Eosinophils have been associated with work demonstrated that pathways involved protection against Haemonchus contortus in the pathogenesis of ovine fasciolosis in sheep both in vitro and in vivo. IL-5 is a included inhibition of macrophage nitric oxide key cytokine in eosinophil induction and production, fibrosis, and antibody isotype activation in vivo. In this study, impairment switching, among others. Transcriptomic of the eosinophil response was attempted changes in PBMC populations following F. by the administration of three subcutaneous hepatica infection in cattle, in which the immunizations of Canaria Hair Breed (CHB) disease phenotype is quite different, have not lambs with recombinant ovine IL-5 (rIL-5ov) yet been examined. Using RNA sequencing prior to infection with H. contortus. Sheep we investigated gene expression changes in vaccinated with rIL-5ov in Quil-A adjuvant, PBMC isolated from 9 non-infected and 11 F.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 35 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

hepatica-experimentally-infected calves at were undertaken: the length of the larvae, the day of infection, at 1 week and at 14 weeks larval survival, response to ivermectin in a post-infection. Longitudinal time-course migration assay, the ability of the larvae to comparisons among groups revealed 21 and exsheath in vitro, and establish and develop to 551 DEGs driven exclusively by F. hepatica the adult stage in young lambs. infection in cattle at acute and chronic stages, respectively. In addition, significant numbers The results for all traits indicate a significant of DEGs related to growth and maturation difference between the host animals, with were detected in both groups at week 14. larvae from specific hosts following a These results show that fewer DEGs at the consistent pattern with the highest or lowest acute stage of infection can be identified in trait results. Compared to larvae from Host cattle, as compared with sheep. Our results 1 the larvae from Host 5 were shorter (741 to reflect the major differences in the disease 692 m, p<0.05), had a longer median survival phenotype between cattle and sheep and (3.6 to 6.4 days), were less susceptible to may reveal pathways to target in vaccine ivermectinµ as indicated by their EC50 (1.2 to development. It also highlight the importance 4.5 M), exsheathed to a lesser degree (83.6 of including an uninfected group in the to 58%), but showed a higher establishment experimental design for transcriptomics- rateµ in the consecutive host (15.2 to 31.4%). based infection time-course studies to control Regarding the survival time, anthelmintic for physiological changes due to growth and susceptibility and establishment rate as development. The significant differences in indicators for fitness, the parasites populating DEGs, immunological pathways, and their Host 5 produced progeny of greater fitness. relevance for the development of anti-liver The findings indicate that the host animal fluke vaccines are discussed. of the parental parasite generation has a significant effect on the parasite progeny.

OA07.03 Host Effects on Haemonchus Contortus Larval Traits OA07.04 Performance of Barbervax® Vaccination in Lactating Merino Ewes Dr Tania Waghorn1, Dr Christian Sauermann1, Dr Kiliana Bekelaar1, Dr Dave Leathwick1 Mr Mulisa Kebeta1, Dr Emma Doyle1, Prof 1Agresearch, Palmerston North, New Zealand Stephen Walkden-Brown1, Dr Brad Hine2, Prof Lewis Kahn1 Nematode parasite control is currently based 1School of Environmental and Rural Science, largely around the use of anthelmintics University of New England, Armidale, however, alternative approaches including Australia, 2CSIRO Animal Livestock Industries, the breeding of animals for resistance Armidale, Australia to nematode infection is based on the capacity of grazing ruminants to mount a Barbervax® vaccination against Haemonchus strong immune response against nematode contortus has been commercially available infection. It has been shown that resistant in Australia since 2014. The optimum use of animals are able to influence the percentage booster vaccination in lambing ewes has not establishment of ingested larvae, the survival been identified. The aim of this experiment rate of adult worms and the fecundity of was to determine the performance adult females. In this work we investigate of Barbervax® in lactating ewes when the influence of host animal on a range of administered one week prior to lambing life-history variables associated with the free- or at lamb marking (seven weeks after living stages of H. contortus. lambing began) to reduce worm burden. The experiment used 400 single bearing Merino A group of 5 lambs was infected with the ewes, previously primed with Barbervax®, same batch of Haemonchus contortus and on a commercial sheep property in the New after patency individual faecal samples were England region of New South Wales. The collected, separately cultured and a series of experiment had a complete 2 x 2 factorial experiments assessing different larval traits design with two treatments administered

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 36 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

pre-lambing [booster vaccination (V) or O. ostertagi infection in cattle does not elicit anthelmintic treatment with Startect® (T) the classic Th2 (anti-inflammatory) paradigm and the same two treatments administered suggesting that an overall mixed response (Th1, at lamb marking time (7 weeks after the Th2, and other types of Th) may dominate. commencement of lambing) providing 4 Following infection, infective third stage larvae treatment combinations of 100 ewes each (L)3s develop into L4s and then to early adults (VV, VT, TV, TT). Taking the start of lambing in the abomasal glands and actively produce as week 0 (W0), body weight and blood excretory-secretory (ES) antigens while causing samples for serological and haematological significant tissue damage in abomasum. Little analysis were conducted at W-1 (prelambing), is known about the regulation of immune W7 (lamb marking) and W18 (weaning). responses at the host-parasite interface during Additional bloods were sampled at W9 for these early and most important phases of the antibody test. Worm egg counts (WEC) were infection. Our research investigated the early taken at lamb marking, six weeks later and at onset of the host immunity. We first determined weaning. The results revealed that pre-lambing the kinetic responses of genes involved in Barbervax® booster vaccination significantly innate immunity, such as Toll like receptors reduced worm egg output at lamb marking (TLRs), cytokines and co-stimulatory markers compared to anthelmintic treatment. Booster in peripheral blood mononuclear cells using vaccination given at lamb marking was not quantitative real time PCR (qRT-PCR). We effective at reducing WEC compared to then examined the influence of O. ostertagi anthelmintic treatment and there was no ES antigens on host innate immune cells, i.e., interaction between the effects of pre-lambing macrophages, by in-vitro cell activation assays, and marking treatments. Body weight and qRT-PCR and immunocytochemistry. Results packed cell volume did not differ between show that upon progression of the disease, the treatment groups throughout the trial. It gene expression of a subset of TLR increased can be concluded that Barbervax® booster indicating active involvement of cellular vaccination at pre-lambing, but not lamb immune response. Additionally, changes in marking time conferred additional protection cell morphology coupled with transcriptional for reproducing ewes treated with an effective changes in TNF and TGF suggest a regulatory short acting anthelmintic at these times. phenotype that is consistent with classical signs of macrophageα activation.β These results demonstrate that the TLR and macrophage OA07.05 Early Host Immune Responses mediated responses are initiated early during to the Cattle Gastrointestinal Nematode infection and may be key to elucidating the Ostertagia Ostertagi complex and intimate relationship between O. ostertagi and its bovine host. Dr. Mariam Bakshi1,2, Dr. Wenbin Tuo1, Dr. Dante Zarlenga1 1United States Department of Agriculture, OA07.06 Abomasum Histomorphometric of Agricultural Research Services, Beltsville, Lambs Infected by Haemonchus Contortus United States, 2Oak Ridge Institute for Science and Fed with Different Protein Levels and Education, Oak Ridge, United States Camila Chaves1, Paula Henriques1, Victor Ostertagia ostertagi is an abomasal mucosa- Nakaguchi1, Isabela Moraes1, Professor dwelling parasite of cattle that imposes Alessandro Amarante2, Luciana Katiki3, significant economic impact on the US cattle Professor Helder Louvandini1 industry. With the rise in drug resistance among 1Centro de Energia Nuclear na Agricultura, nematodes, vaccination is advanced as the Piracicaba, Brazil, 2Universidade Estadual most cost-effective control alternative; however, Paulista, Botucatu, Brazil, 3Instituto de nematode vaccines have met with lackluster Zootecnia, Nova Odessa, Brazil results thus far. Hence, elucidating cellular and molecular mechanisms underlying O. The aim of this study was to evaluate the ostertagi infection is important for developing effects of dietary protein levels (high: 17% efficient vaccines. Reports show that a primary and low: 10%) and Haemonchus contortus

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 37 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

infection in sheep. Twenty four Santa Inês samples were collected from 20 conventional lambs, six months old, randomly distributed in and 19 organic sheep farms with between 70 a factorial arrangment 2x2: high protein control and 250 lactating ewes. The samples were (HPc n = 5), high protein infected (HPi n = 7), pooled into three to five triplets per flock low protein control (LPc n = 5) and low protein on each sampling occasion (in total 270) infected (LPi n = 7). Lambs were infected and the fresh samples were examined by in a single oral dose of 10,000 H. contortus microscopy. Droplet digital (dd)PCR assay L3 larvae. After 42 days of the infection, the was also carried through using DNA extracted animals were slaughter for sample collection from frozen faeces. Most farms (95%) were of abomasum, that was preserved on formalin investigated on two occasions, first samples 10%, embedded in paraffin for cutting which were collected from ewes prior to turn-out were stained by hematoxylin and eosin. Ten and then from lambs after they had been on images from each abomasum were evaluated grass for at least 5 weeks. By microscopy, in 100x lens light microscopy to measured: extra information about the Haemonchus length of the epithelial and glandular layer status was provided on 48% of the sampling (CEG), mucosal muscular thickness (MM), total occasions by including more than two triplets length of mucosal layer (CM); stereological which was the standard method irrespective analysis to calculate density (SD) and surface of flock size applied before 2015 in Sweden. volume (SV). For the variables CEG, CM and At the farm level, detection was further SD, there were effects of diet and infection, enhanced by 13% by replacing microscopy but there was no interaction between them, with the ddPCR assay. There was moderate with lower values for HP compared to LP agreement between the two diagnostic tests and higher for infected compared to controls (Cohens kappa 0.49 ±0.11). On five farms (P<0.05). In MM there was only effect of the Haemonchus was diagnosed by ddPCR only, infection with greater muscle layer for the whereas on one farm the parasite was only infected compared to controls (P<0.05). In SV found with microscopy. Ewes were more often there were no differences between treatments Haemonchus positive than lambs irrespective and no interaction (P>0.05). The HP group of the diagnostic method used indicating that showed thinner abomasal mucosa. The LP and the level of parasite control was acceptable infected groups had thicker mucosa. In the on most farms. Although more ewes were first case to allow a better digestibility of the infected no difference in FEC was observed lower amount of protein and in the second, the between age groups or production systems inflammatory process induced by parasitism (conventional or organic). Combined our led to the thickening of the mucosa, the same results show that Haemonchus infection is occurring with muscular layer with increased widespread but it can be controlled using contractions to eliminate the worms. information obtained with this sampling strategy. In conclusion, we have developed a practical tool for sheep producers to assess OA07.07 Assessment of an Enhanced Haemonchus infection which can be further Sampling Protocol and Use of a ddPCR Assay enhanced by replacing microscopy with for Detection of Haemonchus Contortus on ddPCR. Commercial Sheep Farms

Johan Höglund1, Dr. Safaa T. Elmahalawy1, Dr. Peter Halvarsson1, Dr. Katarina Gustafsson2 1Swedish University of Agricultural Sciences, Uppsala, Sweden, 2Farm and Animal Health Service, Uppsala, Sweden

A novel sampling strategy based on faeces sampling covering approximately 10% of the animals in the flock and with focus on Haemonchus contortus was evaluated. During 76 different sampling occasions 810 individual

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 38 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA08 Tick Disease Transmission OA08.02 Borrelia Burgdorferi Infection Modifies Protein Content in Ixodes Scapularis July 8, 2019, 13:30 - 15:30 Nymph Tick Saliva Breakout Room 3, Hall of Ideas F&I, Level 4 Tae Kwon Kim1, Lucas Tirloni1, Jolene Diedrich2, John Yates2, Albert Mulenga1 OA08.01 Immunogens Expressed by Tick 1Texas A&m University, College Of Veterinary Transmitted Borrelia Burgdorferi Medicine & Biomedical Sciences, College Station, United States, 2Molecular Medicine Emily Bencosme Cuevas1, Tae Kwon Kim1, and Neurobiology, The Scripps Research Lucas Tirloni1, John Yates2, Jolene Diedrich2, Institute, LaJolla, United States Albert Mulenga1 1College of veterinary medicine and Majority of human vector borne diseases biomedical sciences, Department of in the US is primarily tick borne. Lyme Veterinary Pathobiology, Texas A&M disease (LD) caused by spirochets, Borrelia University, College Station, United States, burgdorferi (Bb) and B. mayonii is the most 2Molecular Medicine and Neurobiology,The common human tick-borne disease (TBD) in Scripps Research Institute, LaJolla, United the US. LD is primarily associated with Ixodes States scapularis nymph tick bites. There is evidence that proteins in saliva of I. scapularis nymphs Majority of human vector borne diseases in play significant roles in transmission of LD the US is primarily transmitted by hard ticks. spirochetes. However molecular identities Lyme disease (LD) caused by spirochetes, of I. scapularis nymph ticks have not been Borrelia burgdorferi (Bb) and the recently determined. The goal of this study was to described B. mayonii, is the most common identify proteins that Bb-infected I. scapularis human vector borne disease in the US. These nymph inject into the host during feeding. LD spirochetes are transmitted by the deer We successfully used LC-MS/MS sequencing tick, Ixodes scapularis during feeding. Nearly to identify proteins that uninfected and Bb 30,000 LD cases are reported to the US CDC infected I. scapularis nymphs sequentially each year with 300,000 cases estimated. In inject into the host every 12 h during feeding. order to develop effective methods to prevent Overall, we have identified an additional more LD, understanding the molecular basis of how than 100 proteins in saliva of Bb-infected I. LD spirochetes interact with their tick vectors scapularis nymphs that were not found in and vertebrate hosts is needed. The goal of uninfected ticks. These data for the first time this study was to identify Bb immunogens reveals molecular identities of tick saliva that are expressed in both its arthropod (tick) proteins that might play important roles in and vertebrate stages. Antibodies to tick transmission of Bb. transmitted Bb from rabbits that were infested by Bb-infected I. scapularis nymphs was used to probe western blots of cultured Bb OA08.03 Prevention of Babesia Rossi protein extracts to identify Bb immunogens. Transimission by Haemaphysalis Elliptica on More than 20 immunodominant protein bands Dogs Treated With Nexgard® were excised and processed for LC-MS/ MS sequencing and identification. Further, Prof. Frederic Beugnet1, Wilfried Lebon1, immune sera to tick transmitted Bb will be Christa De Vos2 used to immuno-screen a Bb-genomic DNA 1Boehringer-ingelheim, Lyon, France, 2ClinVet, phage display library. We discuss our findings Bloemfontein, South Africa with reference to understanding the molecular basis of Bb transmission and progress toward The objective of this study was to determine identification of vaccine antigens against LD. the efficacy of afoxolaner (NexGard®) to prevent Babesia rossi transmission by Haemaphysalis elliptica ticks on dogs. The study was conducted with 3 groups of 7 dogs: G1 and G2 were negative control groups,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 39 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

infested on Day 2 and Day 28, respectively. (KFD) in the year 1957 from a small pocket G3 dogs were treated group on day 0 and of Karnataka state in India, the prevalence infested by 50 Haemaphysalis elliptica ticks of virus in human subject has been reported on Day 2 and on Day 28. These ticks were from adjoining states till 2019. Different originated from nymphs having fed on B. species of ticks have been incriminated rossi infected dogs. Infection rate of the ticks as possible vector of the pathogen. After was PCR assessed and it was 12.8% at Day 2 the recent reports of monkey’s death from (from a batch of 40 ticks) and 6% at Day 28 local region of Shimoga district, Karnataka, (from a batch of 50 ticks). All these dogs had on recommendation of local health care a weekly veterinary examination and a daily professionals, the ticks were collected visual observation. On Days 0, 7, 14, 21, 28, from the local cattle breed, malnad gidda, 35, 42, 49 and 56, and in case of suspicion of and forest vegetation. The people living fever (i.e. body temperatures > 39.4 °C) or adjacent to the forest region of Shimoga any other babesiosis sign, blood samples were are maintaining this breed and leaving the collected for blood smears and PCR detection animals in the forest for grazing. A detail of Babesia rossi DNA. The B. rossi infection history was collected from the people who rate in the untreated control group challenged are living in that area and surrounding forest on Day 2 (group 1) was 100% (6/6). It was regions revealed several cases of skin rashes 57.1% (4/7) in the control group challenged on due to tick bites. The ticks were processed Day 28 (group 2). The rate of B. rossi infection and morphologically identified as nymphal in the afoxolaner treated groups was 0% (0/7) and adult stages of Haemaphysalis spinigera. after the two tick challenges and until Day 56. The animal owners reported tick bite which The ticks were thumb counted at 48 hours causes itching, erythematous, popular and counted and removed at 144 hours after lesions on the hands, neck and leg region. each infestation. The untreated control groups The suspected samples from human, dead had an arithmetic mean of ticks ranging monkeys and ticks were submitted to National from 23.8 to 26.8 on these assessment days. Institute of Virology (NIV), Pune, the nodal Based on arithmetic mean values of live ticks, Indian agency for case confirmation of KFD. NexGard® provided 100% acaricidal efficacy The report confirms the presence of KFD virus against H. elliptica ticks on all assessments. in the submitted samples. The health care This study demonstrated that NexGard® professionals were suggested to carry out protected dogs against infection by B. rossi immediate vaccination in order to prevent the for a month. outbreak of KFD. The peoples living adjacent to the forest were advised to apply tick repellent oil before entering in to the forest. OA08.04 Haemaphysalis Spinigera a Vector The spraying of acaricide (Malathion) in the for Outbreak of Kyasanur Forest Disease in outbreak areas was carried out to manage the Shimoga: A Malnad Region of Karnataka tick population.

Dr Ananda Konanur Javaregowda1, Dr Kiran SK2, Dr Srikant Ghosh3 OA08.05 No Evidence of Bartonella Henselae 1Department Of Veterinary Parasitology, Transmission From Larva to Nymph Stages of Karnataka Veterinary, Animal And Fisheries Rhipicephalus Sanguineus by Using Artificial Sciences University (b), Veterinary C, Membrane Feeding System Shimoga, India, 2Department of Health and Medical Sciences, Shimoga, India, 3Division Wittawat Wechtaisong1, Sarah Bonnet2, Yi- of Veterinary Parasitology, Indian Veterinary Yang Lien1, Yi-Lun Tsai1 Research Institute, Izathnagar, India 1Department of Veterinary Medicine, College of Veterinary Medicine, National Pingtung Ticks are next to mosquitoes which are University of Science and Technology, involved in the transmission of variety of Pingtung, Taiwan, 2UMR BIPAR, INRA, Ecole disease causing pathogens of public health Vétérinaire d’Alfort, ANSES, Université Paris- and veterinary importance. After the first Est, Maisons-Alfort, France report of fatal Kyasanur Forest disease

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 40 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Bartonella spp. are fastidious gram-negative globally. Most of these pathogens are bacteria which confirmed to be or as transmitted through the vector’s saliva potential to be human and animal pathogens. during blood feeding. Ticks transmit B. henselae are the cause of cat scratch bacteria, viruses, and protozoans, and in disease (CSD) in humans, transmitted by some parts of the world are more important cat scratch with claws contaminated with than mosquitoes as vectors of zoonotic Bartonella positive flea feces. There has been diseases affecting humans. The southern considerable interest in ticks as potential cattle fever tick, Rhipicephalus microplus, vectors for Bartonella spp. The aim of the is the most economically important present study was to clarify the vector ectoparasite of cattle worldwide. Our previous competency of R. sanguineus ticks for B. research documenting the presence of henselae transmission. An artificial membrane acetylcholinesterase (AChE) in the saliva of R. feeding system was used to infect ticks with microplus proposed this enzymatic activity B. henselae and to evaluate bacteria trans- could: 1) decrease the toxicity of acetylcholine stadial transmission within tick developmental in the blood meal, 2) modulate host immune stages. 400 larvae were engorged with responses to facilitate tick parasitism, and 3) Bartonella-infected blood, and, after molting, facilitate salivary assisted transmission (SAT). 60 nymphs were engorged on non-infected We hypothesized further that if tick salivary blood. Blood culture from a feeder was then acetylcholinesterase participates in pathogen performed at day 4 of nymph feeding. 200 SAT then similar enzymatic activity would larvae and 40 nymphs fed with non-infected be present in the saliva of other arthropod blood were considered as the control groups. vectors. This reports includes evidence of In both experimental and control groups, PCR AChE-like activity in the saliva of several were applied on various samples, including ticks, mosquitoes, sand flies, and apparently blood from a feeder, pooled engorged larvae, biting midges. Salivary AChE-like activity pooled semi-engorged larvae, pooled larva was not detected in the horn fly, stable fly, feces, pooled nymphs and pooled nymph or house fly. Salivary cholinesterase (ChE) feces. The PCR results showed that all samples activities detected in arthropod vectors were negative for Bartonella spp. except exhibited Michaelis-Menten KM values lower blood sample from the experimental feeder. In than the KM value for bovine serum AChE. addition, no bacterial colonies were found after A lower KM value is indicative of higher 14 days of incubation from blood culture after affinity for substrate, and is consistent with feeding of nymphs infected as larvae. These a hypothesized role in localized depletion of results indicated no evidence of B. henselae host tissue acetylcholine and modulation of infection of larvae after tick feeding on infected host immune responses at the vector bite site blood through membrane feeding and no that might favor hematophagy, which could transstadial transmission from larva to nymph enable infection by vector-borne pathogens. stage of R. sanguineus. In future, transstadial transmission of B. henselae from nymph to adult and transovarial transmission will be evaluated. OA08.07 Trends in Canine Infection with Borrelia Burgdorferi and Anaplasma Phagocytophilum in the Eastern United OA08.06 Potential Involvement of Salivary States: 2010–2017 Cholinesterase Activity in Arthropod Vector- Borne Disease Transmission Bhagya Kulasooriya1, Dr. Susan Little1, Dr. Mark Payton2, Melissa Beall3, Jennifer Braff3, Dr. Kevin Temeyer1, Kristie Schlechte1, Dr. Jesse Buch3, Andrew Knupp3 Adalberto Perez de Leon1 1Department of Veterinary Pathobiology, 1USDA-ARS KNIPLING-BUSHLAND US Oklahoma State University, Stillwater, United Livestock Insects Research Laboratory, States, 2Department of Statistics, Oklahoma Kerrville, United States State University, Stillwater, United States, 3IDEXX laboratories Inc, Westbrook, United Arthropod vectors transmit pathogens States responsible for 17% of all infectious diseases

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 41 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Borrelia burgdorferi and Anaplasma Shafy6, Dr Raafat Shaapan7 phagocytophilum are tick-borne infections 1Shaqra University, Ad-dawadimi , Saudi transmitted by Ixodes scapularis in the eastern Arabia, 2King Faisal University, Al-Ahsa , Saudi United States; both agents cause disease in Arabia, 3King Saud University, Riyadh, Saudi dogs and people. Cochran Armitage trend test Arabia, 4Damietta University, Damietta , Egypt, (P<0.0001) was used to evaluate changes in 5University of Khartoum, Khartoum, Sudan, annual percent positive results for antibodies 6National Research Centre, Dokki, Egypt, to B. burgdorferi and Anaplasma spp. in 7National Research Centre, Dokki, Egypt approximately 20 million canine tests in 25 states in the eastern United States. From 2010 Hard ticks are among the most important to 2017, a decreasing trend in percent positive blood sucking arthropods that transmit test results for B. burgdorferi that ranged pathogens to humans and animals. This study from 16.7% to 48.6% was evident in 8 states was designed to determine the prevalence, along the mid-Atlantic coast from Virginia to map the geographical distribution and New Hampshire, and in Wisconsin. In contrast, examine the seasonal activity of hard tick a continued increasing trend was evident in species infesting the most common domestic 7 northeastern and midwestern states where and wild mammals in various districts of Lyme borreliosis is endemic or emerging, as Riyadh Province, Saudi Arabia during the well as in 3 southern states where endemicity period of January to December 2017. A has not yet been established, while no overall total of 11,587 hard ticks were collected trend was evident in Vermont or 5 other from the bodies of 8,435 animals belonging midwestern states. Similarly, percent positive to 18 different mammalian species. The test results for A. phagocytophilum showed a ticks were preserved in 70% alcohol, and significant, although smaller, decreasing trend microscopy was used to identify species. in 6 states along the mid-Atlantic coast from Two genera, Hyalomma and Rhipicephalus Virginia to Connecticut and Rhode Island, as were identified, comprising ten species of well as in most states in the Midwest. However, hard ticks, with Hyalomma comprising 68.3% a strong increasing trend was evident in and Rhipicephalus comprising 31.7% of Massachusetts and northern New England as species. The most common species found on well as Appalachian regions in West Virginia domestic mammalian hosts was Hyalomma and Pennsylvania. Although percent positive dromedarii (Koch,1844) (39.9%) followed test results continued to increase in regions by Rhipicephalus turanicus (Pomerantsev, where Lyme disease and anaplasmosis are Matikashvili & Lotosky,1936) (34.9%); while more newly endemic, the significant decrease on wild mammalian hosts Rhipicephalus evident in other areas over the 8-year period sanguineus (Latreille 1806) was by far the considered was unexpected. This change may most prevalent species (83.0%). These ticks reflect the combined positive influence of were most abundant from May through canine vaccination, tick control, and routine July (36.0%) in the studied areas, and tick testing of dogs in regions where these abundance differed significantly among infections have long been endemic. Analysis seasons (t=7.854, df=11, P=0.000). Regression of trends in canine test results for tick-borne analysis for tick intensity was highly infections continues to be a valuable tool to significant on domestic animals (F = 5.365, understand relative geographic and temporal P=0.006), while it was not significant on risk for these zoonotic agents. wild animals (F = 2.494, P=0.088). However, this study provides new data on the current status of ticks for human and animal health OA08.08 Species Diversity and Seasonal service managers, as well as for governmental Distribution of Hard Ticks (Acari: Ixodidae) authorities, to gain a better understanding Infesting Mammalian Hosts in Various of the occurrence of hard ticks infesting Districts of Riyadh Province, Saudi Arabia mammalian hosts in Riyadh Province, Saudi Arabia, which can help improve prevention Dr Abdullah Alanazi1, Prof Hamdan Al- and control of tick-borne diseases, especially Mohammed2, Prof Mohamed Alyousif3, Prof during outbreaks. Asraf Said4, Dr Bashir Salim5, Dr Sobhy Abdel-

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 42 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA09 Wildlife Parasites OA09.02 Effects of Sarcoptic Mange on American Black Bear Activity and Movement July 8, 2019, 13:30 - 15:30 Patterns in Pennsylvania Breakout Room 4, Hall of Ideas G&J, Level 4 Hannah Greenberg1, Dr. Erika Machtinger1, Mark Ternent2, Dr. Justin Brown3, Dr. Jennifer OA09.01 Ticks, Keds, and Citizen Scientists: Murrow4 Working With Hunters to Better Understand 1Department of Entomology, The Pennsylvania Wildlife Parasites State University, State College, United States, 2Bureau of Wildlife Management, Pennsylvania Dr. Erika Machtinger1, Dr. Michael Skvarla1, Dr. Game Commission, Harrisburg, United States, Graham Hickling2 3Department of Veterinary and Biomedical 1Pennsylvania State University, State College, Sciences, The Pennsylvania State University, United States, 2University of Tennessee, State College, United States, 4Department Knoxville, United States of Environmental Science and Technology, University of Maryland, College Park, United Ticks are frequently encountered by hunters, States outdoor enthusiasts, and rural residents and can carry pathogens that cause diseases such Since 1991, sarcoptic mange has increased as Lyme disease, Powassan virus Disease, and in American black bears (Ursus americanus) Anaplasmosis. Deer keds are less well-known in Pennsylvania, both in incidence and ectoparasites of Cervidae that occasionally geographic distribution and has recently bite humans. Recent small-scale studies expanded into surrounding states. This have sequenced pathogens traditionally skin disease is caused by a parasitic mite, considered to be tick-vectored from keds, Sarcoptes scabiei that burrows under the but pathogen distribution, diversity, and host’s skin as part of its life-cycle. This causes occurrence of co-infections in this group are a hypersensitivity reaction in the host that unknown. In addition, despite their actual can lead to significant skin disease, as well and potential medical importance, data as atypical behavior, altered activity patterns on the distribution of ticks, keds, and tick- causing health declines, substantial weight borne diseases in Pennsylvania is lacking. loss, as well as secondary infections. In some This information, along with basic vector cases, these effects can be severe enough to education, is critical to mitigate potential cause mortality. This study aims to characterize disease risks faced by recreational hunters changes in movement, resource utilization, and their animals. To address this, in 2018 we and health parameters in bears with sarcoptic developed the Pennsylvania Parasite Hunters mange. Over a two-year period, 36 bears will (www.paparasitehunters.com) citizen science be fitted with GPS-collars and followed for project to engage hunters in surveillance five years. Between April – October 2018, 28 of ticks and keds in the State. Participating bears were collared. Bears were collared in hunters ordered postage-paid kits to collect triplicates according to gender and similar specimens from harvested deer. We had a habitats, consisting of one bear without mange return rate of 28.4% representing 30 counties (i.e. healthy control), one bear with confirmed in Pennsylvania. In addition, we sampled 85 sarcoptic mange treated with a single injection deer from 24 counties at participating deer of ivermectin, and one bear with confirmed processing locations for a combined coverage sarcoptic mange that was untreated. Data were of 43 of 67 Pennsylvania counties and a total analyzed in R Studio to study differences in of 753 ked and 1,048 tick specimens. Overall, movement and activity patterns between these the pilot was a successful and efficient way three groups. These data will be used to inform to survey wildlife parasites from a wide management decisions on whether bears with geographic area and participants were sarcoptic mange should be treated, will define engaged in the study. Pathogen status and the efficacy of treatment with a single dose of multi-state tick and ked behavior observations ivermectin, and will contribute to the present will be discussed. understanding of the effects of sarcoptic mange in black bear populations.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 43 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA09.03 First Report of Cryptosporidium OA09.04 Mass Mortality in Endangered Parvum Subtype IIaA16G3R1 in Cervids Pinna Nobilis (Linnaeus 1758) Fan Mussels Associated with Haplosporidium Pinnae Dr. Weslen Teixeira1, Dr. Márcio de Oliveira3, Dr. Pedro Peres3, Ms/ Walter Bertequini Nagata2, Dr. Rossella Panarese1, Dr. Perla Tedesco2, Dr. Bruna Nicoleti Santana2, Ms. Bruno Oliveira2, Giovanni Chimienti1, Professor Maria Stefania Dr. José Duarte3, Dr. Marcelo Meireles2, Dr. Latrofa1, Professor Francesco Quaglio3, Dr. Welber Lopes1, Unesp Katia Bresciani2 Andrea Gustinelli2, Professor Angelo Tursi1, 1Universidade Federal de Goiás, Goiânia, Professor Domenico Otranto1 Brazil, 2Universidade Estadual Paulista 1University of Bari, Bari, Italy, 2University of (UNESP), Faculdade de Medicina Veterinária Bologna, Bologna, Italy, 3University of Padova, de Araçatuba, São Paulo, Brasil., Araçatuba, Padova, Italy Brasil, 3Universidade Estadual Paulista (UNESP), Faculdade de Ciências Agrárias e The fan mussel Pinna nobilis (Linnaeus 1758) Veterinárias de Jaboticabal, São Paulo, Brasil, is an endemic bivalve of the Mediterranean Jaboticabal, Brasil basin, protected by international legislation as an endangered species. In the early We characterized genetically the infections by summer of 2018, a mass mortality event Cryptosporidium in Mazama gouazoubira. By a (MME) of P. nobilis was recorded in the non-invasive harvest methodology using trained Gulf of Taranto (Southern Italy, Ionian Sea). sniffer dogs to locate fecal samples of cervids, Moribund specimens of P. nobilis were 642 fecal samples were obtained from six collected by scuba divers and processed Brazilian localities. The cervid species responsible by bacteriological, parasitological, for the excretion of each faecal sample were histopathological and molecular analyses identified by the polymerase chain reaction to investigate the causes of such a MME. (PCR) performed from the genomic DNA Different developmental stages (i.e., previously extracted. From this identification, plasmodia, spores and sporocystis) of 437 fecal samples of M. gouazoubira were Haplosporidium spp. were observed during selected for research of Cryptosporidium spp. the histological analysis in the epithelium performed through negative staining with and in the lumen of the digestive tubules, malachite green and polymerase chain reaction where mature spores occurred either free (nPCR) followed by sequencing the amplified or in sporocysts. The spores presented an products. In the samples that were diagnosed operculum and an ovoid shape measuring 4.4 the presence of parasite species with zoonotic m (±0.232) in length and 3.6 m (± 0.233) potential, genotyping was also performed in width. The BLAST analysis of 18SrRNA using nPCR with the subunit of GP60 gene. sequenceµ revealed a high nucleotideµ similarity Statistical analysis consisted of the Fisher exact (99%) with the reference sequence of test to verify the association of the presence of Haplosporidium pinnae available in GenBank the enteroparasite in relation to the presence database. Accordingly, at the phylogenetic of cattle in each locality, and the McNemar analysis, 18SrRNA sequence was clustered tests and Kappa correlation coefficient used as a paraphyletic clade with the reference to compare the results obtained between sequence of H. pinnae, excluding other the two diagnostic techniques. In the fecal haplosporidians (i.e., Bonamia and Minchinia samples of M. gouazoubira the occurrences genera). Based on data reported, H. pinnae of Cryptosporidium were diagnosed in was the causative agent of MME in the 1.6% (7/437) and 1.1% (5/437), respectively, populations of P. nobilis sampled in the through nPCR and microscopy. C. parvum Ionian Sea, where the conservation of this was diagnosed in 100% (7/7) of the samples endangered species is heavily threatened submitted to sequencing (18S gene). The by such a protozoan infection. Further IIaA16G3R1 subtype was diagnosed in five of the investigations should regard the life cycle of C. parvum samples submitted to genotyping H. pinnae, in order to reduce the pathogen (GP60 gene). This is the first world report of spreading and to mitigate the burden of the C. parvum in M. gouazoubira and subtype disease where P. nobilis is facing the risk of IIaA16G3R1 in cervids. extinction.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 44 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA09.05 Unravelling the Parasites of Deer OA09.06 First Record of Possible Life Cycle Stages of a Hepatozoon Blood Parasite Alex Chambers1, Mr Paul Candy1, Dr Dave Species (ApicomplexFirst Record of Leathwick1 Possible Life Cycle Stages of a Hepatozoon 1AgResearch Ltd, Palmerston North, New Blood Parasite Species (Apicomplexa: Zealand Adeleorina: Hepatozoidae) in an Ixodes Tick (Arthropoda: Ixodida: Ixodidae) and Parasites are considered to be the most an African leopard, Panthera pardus pardus important animal health issue for farmed (Linnaeus, 1758) deer in New Zealand. Deer parasites have seldom been studied in any depth, and deer Mrs. Michelle Van As1,2, Dr. Johann van As2, are host to some unusual nematode species. Prof. Oriel Thekisoe1, Prof. Nico Smit1 A better understanding of the infective 1North West University, Potchefstroom, species present, and their trends, will help South Africa, 2University of the Free State, inform host-parasite life-cycles, in addition to Phuthaditjhaba, South Africa developing more effective control programs. Our aim is to describe the longitudinal Intracellular apicomplexan haemoparasites seasonality of egg/ larval output in deer. from the genus Hepatozoon Miller, 1908 have been described from a wide range of Five deer farms were selected on the basis vertebrate hosts, including wild carnivores. of location, control practices and willingness Reports on these haemoparasites from the to participate in the study. Faecal samples African Leopard, Panthera pardus pardus were collected from pasture by group (hinds, (Linnaeus, 1758), are scarce and generally stags and fawns) every 3-4 weeks. Individual non-specific. Furtheremore, information on Strongyle faecal egg and lungworm counts the mode of transmission and descriptions of were performed, and the remaining faeces life cycle stages in infected vectors remains pooled by group per farm and cultured to third relatively rare. The aim of this study was to stage infective larvae (L3). 2000-3000 L3 from explore the role of ticks as potential vectors each group in time by farm were speciated by of a Hepatozoon species infecting African deep amplicon sequencing methods. leopards in South Africa. Peripheral blood samples and engorged ticks were collected The results show that farmed deer in New from five wild leopards, three females and two Zealand have very low year-round faecal males, while under sedation. Giemsa stained egg counts, in comparison to other farmed smears of peripheral blood were methodically ruminants, with little to no seasonal patterns. screened for Hepatozoon gamont stages, Whilst lungworm infections are thought to both extra- and intraleukocytic. Engorged be the biggest threat to deer farming, the ticks collected from infected leopards were counts were very low for all farms and groups. kept alive in a fasting state for seven days The speciation data will be presented, once before being squashed on clean microscope analysis is completed, at the conference. This slides, stained with Giemsa solution, and study will be the first to identify nematode screened for various possible developmental parasites in New Zealand deer by next stages. Sporogonic stages, including generation sequencing, and will inform the microgametes, immature and mature oocysts seasonal pasture contamination and assist and infective sporozoites, were observed with developing control strategies. in a tick (Ixodes sp.) collected from a male leopard infected with mature and immature There is a disparity in control programs gamont stages of a Hepatozoon species. between the farms, generally animals over the Developmental stages were photographed, ages of 18 months are not drenched, younger differentiated and measured with ImageJ animals can be drenched up to 10 times. As software. This is the first report on the the New Zealand deer industry continues to characteristics of different developmental grow, there is an increasing need to better stages of a feline species of Hepatozoon in understand the impact of management both its potential tick vector and African decisions on sustainable parasite control. leopard host.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 45 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA09.07 Sarcosporidiosis: An Emerging OA10 New Tools and Big Data Disease in Yaks (Bos Grunniens) for Evaluating Intestinal Parasite Infections in Companion Animals Dr Khalid Mehmood1,2, Dr Kun Li1, Dr Houqiang Luo3, Dr Hui Zhang1, Dr Muhammad Shahzad2, July 8, 2019, 13:30 - 15:30 4 Dr Rao Zahid Abbas Breakout Room 5, Meeting Rooms KLOP, Level 4 1College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, People’s Republic of China, 2University College OA10.01 A Short Review of the 100 Years of of Veterinary & Animal Sciences, Islamia Methods Other Than the Direct Smear for University of Bahawalpu , Bahawalpur, Detecting Parasites in Feces Pakistan, 3College of Animal Science, Wenzhou Vocational College of Science and Professor Dwight D Bowman1 Technology, Wenzhou, People’s Republic of 1Cornell University, ITHACA, United States China, 4Department of Parasitology, Faculty of Veterinary Science, University of Agriculture, In 1918, US Army Majors Kofoid and Barber Faisalabad, Pakistan described a brine flotation method they used to examine 100,000 stool samples from The sarcocystosis is caused by a coccidian service men under diverse field conditions. intracellular protozoan belonging to genus A skilled examiner could perform 150 to Sarcocystis (Apicomplexa: Sarcocystidae: 250 examinations daily, and a staff of 15-20 Sarcocys tinae). Sarcocystosis is a world-wide examiners could process from 2,000 to 3,000 distributed parasitic zoonosis. However, scarce samples daily. The authors stated that “The ova information is available about the infection of of parasites such as Ancylostoma duodenale, Sarcocystosis in yaks. Herein, we conducted Necator americanus, Ascaris lumbricoides, this study to survey the seroprevalence Oxyuris vermicularis, Trichuris trichiura, Taenia of Sarcocystosis in yaks on the Qinghai saginata, Taenia solium, Hymenolepis nana, Tibetan plateau (QTP). A total 2549 serum Hymenolepis diminuta, and Dipylidium caninum samples were obtained during 2011 to 2017, and of trematodes are floated up by the brine which were assayed by piloting commercial into the surface layer of the pool without ELISA kits. The results revealed that the distortion or noticeable change in appearance overall seroprevalence of Sarcosporidiosis during the usual period of examination. Cysts in QTP yaks was 0.90% (95 CI: 0.6-1.4). The of Endameba coli and histolytica and of Giardia seroprevalence was ranged from 0.20% (95 intestinalis are also floated up. Since the ova are CI: 0-1.1) to 1.67% (95 CI: 0.8-3.0) in yaks at the surface, it is not advisable or necessary in different areas. The seroprevalence was to use a cover glass.” Additional methods have 0.73% (95 CI: 0.3-1.4) in male yaks and 0.06% been developed and compared for the past (95 CI: 0.6-1.8) in female yaks. In different century, including the Stoll dilution method, ages, the seroprevalence were ranged from the alternate flotation solutions, improved 0 (95 CI: 0-1.4) to 1.47% (95 CI: 0.6-3.0). In methods with centrifugation, sedimentation different years, the seroprevalence were methods, the McMaster slide, various stationary ranged from 0 (95 CI: 0.-1.4) to 1.86% (95 flotation devices, centrifugal flotation devices CI: 0.7-4.0). In the current study, risk factors (the FLOTAC and StatSpin Ovatube), the Kato of region and age were revealed to be the thick smear, the calibrated direct smear, long- obvious influencing risk factors by piloting distance reading of capture images (FecPak), conditional step-wise logistic regression. and potential video recognition software, and The current study herein first found the many others. The fist major methodologic emerging infection of Sarcosporidiosis in change in veterinary medicine relative to yaks from high plateaus, which contributes parasite detection came with the development to outline the epidemiological scenario of and launch of an assay that could readily detect Sarcosporidiosis in yaks in China. Moreover, the presence of cyst wall antigens of Giardia our findings highlight the urgent need of in fecal samples. This discovery and other Sarcosporidiosis studies in yaks on the QTP detection methods based on rapid immunologic for more comprehensive level. and molecular methods may provide significant

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 46 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

changes in the future as to how fecal samples in contrast, hookworms were most common are examined. This talk will review the past as in the South (3.0%), while whipworms were we prepare to enter the second century of fecal most common in the South and Midwest puddling. (0.7%). Intestinal parasites, some with zoonotic implications, remain important in pet dogs although prevalence of infection is OA10.02 Prevalence of Intestinal Parasites in dramatically reduced compared to that seen Pet Dogs in the United States, 2016–2018 in dogs not receiving routine veterinary care.

Prof. Susan Little1, Donald Szlosek2, DJ McCrann2, Donald Martin2, Melissa Beall2, Troy OA10.03 IDEXX Fecal Antigen Technology: Goddu2 Fecal Dx® Tests 1Oklahoma State University, Stillwater, United States, 2IDEXX Laboratories, Inc., Westbrook, Dr. David Elsemore1 United States 1Idexx Laboratories, Inc., Westbrook, United States To determine the prevalence of parasites in fecal samples from pet dogs in the continental The Fecal Dx® panel (antigen detection of United States and evaluate differences due ascarids, hookworms, and whipworm) have to age and geographic origin, results from been on market since 2016. The antigen tests 4,652,566 zinc sulfate centrifugal flotation detect excreted or secreted proteins from the fecal examinations conducted at IDEXX young adult to adult stages of each nematode. Laboratories from January 2016 to June The discrete protein markers from Toxocara 2018 were randomly selected for analysis. canis, Ancylostoma caninum, and Trichuris Results were grouped by age class and vulpis are not associated with reproduction or region and analyzed by Chi square and one- with eggs and thus provide a novel window way ANOVA with significance assigned at into infection in dogs and cats. Experimental P<0.0001. Parasites identified included Giardia infections with the canonical species (3.3%), hookworms (1.9%), Eimeria spp. demonstrate the ability to detect prepatent (1.6%), Cystoisospora spp. (1.6%), ascarids periods which vary for each nematode life (1.5%), and whipworms (0.6%). Giardia and cycle— as soon as 7 days for T. canis, 14 days Cystoisospora spp. were commonly identified for A. caninum, and 46 days for T. vulpis. in young pups (2–6 months; 12.8% and 6.9%, Conservation of the protein markers within respectively) but prevalence significantly each group of nematodes allows breadth declined with age (>3 years; 0.2–0.8%). In of detection beyond the nematode species contrast, detection of Eimeria spp. remained used to design the assay: ascarid detection fairly constant (1.1–1.7%) throughout all adult includes T. cati, Baylisascaris procyonis, age classes (>1 year); by 1–2 years of age, and Toxoascaris leonina; hookworm species Eimeria spp. was the most common protozoa include A. tubaeforme, A. braziliense, and detected. Giardia and Cystosiospora spp. Uncinaria stenocephala; and the whipworm were most commonly identified in the West test detects T. felis. Testing at IDEXX Reference (5.8% and 2.7%, respectively), while Eimeria Laboratories has resulted in millions of records was most common in the Northeast (2.3%). that contain both flotation and antigen results. Ascarids were most commonly identified in Comparison of egg flotation and the Fecal Dx® young (5.9%) and older pups (7–12 months; panel results shows excellent agreement when 2.1%), and prevalence significantly declined egg observations are recorded and when no with age. Older pups were most likely to be egg is observed (96.8% to 99%). Disagreement shedding hookworm (3.1%) and whipworm between egg observation and antigen (1.5%) eggs; prevalence of hookworms did result ranges from 1-3.2% and may reflect not fall below 1% until >7 years of age, and fundamental differences between egg flotation whipworms reached its lowest prevalence and detection of nematode coproantigens. level >5 years of age. Ascarids were most common in the Northeast, Midwest, and West (1.6%) and least common in the South (1.1%);

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 47 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA10.04 Agreement of Fecal Antigen Test OA10.05 What Is Going On With Trichuris in Results (Fecal Dx®) to Egg Observations Is Cats in Florida? Influenced by Host and Parasite Parameters— Insights From a Large Field Population of Jennifer Ketzis1, Jinming Geng2, David Dogs Elsemore2 1Ross University School Of Veterinary Dr. David Elsemore1 Medicine, Basseterre, Saint Kitts and Nevis, 1Idexx Laboratories, Inc., Westbrook, United 2IDEXX Laboratories, Inc., Westbrook, United States States

A data set of 56,512 canine egg positive Data from feral cats in the Miami, Florida (ascarid, hookworm, or whipworm) samples region in 2014 indicated a high occurrence with accompanying Fecal Dx® antigen of Trichuris (>30%). To confirm this level and test results was collected providing an determine if the same level could be found opportunity to explore patterns of antigen in other regions of Florida, 143 fecal samples agreement to egg observations in pet dogs. from feral cats (estimated to be 4 months of Various host and parasite parameters could age or older) in the Miami region, northwest influence the agreement of antigen to egg. and northeast of Miami and in central Florida Factors that favor patent infections should were collected in August and October 2018 maximize agreement (egg positive and under an approved IACUC protocol. Samples antigen positive) compared to conditions were analyzed using the IDEXX Fecal Dx® that disfavor patency (egg positive but antigen test for whipworm ELISA and antigen negative). To test this idea, antigen centrifugation with zinc sulfate (spg 1.25). to egg agreement for 21,899 Toxocara spp. Of the 143 samples, 78 (55%) of the cats had egg positive samples was examined by dog at least one parasite infection and 39 (27%) age group. A significant drop in agreement had multiple infections. The most common was seen in older dogs consistent with infection was hookworms (Ancylostoma sp. abatement of tracheal-migration in older based on location) with 64 (45%) of the cats dogs. This trend was not observed for dogs positive. Twenty six cats (18%) were positive positive for Ancylostoma caninum (n = for Trichuris sp.: 17 by ELISA and flotation; 8 12,756). Antigen to egg agreement can also by ELISA only; and one positive by flotation be surveyed across data sets where the only. Of these Trichuris positive cats, 24 were dogs were separated into cohorts based on from the greater Miami region. Occurrence co-observation of pseudoparasites such as among the tested population from Miami was Eimeria, Anoplocephela spp. or strongyle 42% (24 of 57) while in all other regions was eggs. Antigen to egg agreement should be <3% (2 of 86). Cats from one shelter might maximized in dogs without a pseudoparasite have been treated; excluding these cats, the co-observation (patent infection more likely) occurrence of Trichuris outside of the Miami as compared to dogs with a pseudoparasite greater region was <2% (1 of 51). While other observation (host behavior(s)/environmental parasite infections were higher in cats from conditions that favor spurious eggs). the Miami greater region, the difference in Significant differences were found for prevalence was not as extreme. For example, Toxocara spp (n = 25,840)., A. caninum (n 56% (34 of 57) of the cats were positive for = 19,300), and Trichuris vulpis (n = 8,028) hookworm in the Miami greater region and egg positive dogs between cohorts with and 46% (28 of 51) from all other areas excluding without a pseudoparasite co-observation. the suspected treated cats (35% (30 of 86) The antigen to egg agreement pattern is including the suspected treated cats). The also different for other species of ascarids reason for the high occurrence of Trichuris or hookworms. Baylisascaris procyonis (n = in the Miami greater region is unclear and 57) and Uncinaria stenocephala (n = 1,970) requires further investigation. antigen to egg agreement is significantly lower than observed for T. canis or A. caninum.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 48 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA10.06 Further Comparison of inferior fecal technique and should not be Centrifugation Versus Passive Fecal Flotation considered as a reliable diagnostic test in for the Recovery of Toxocara Canis, Trichuris practice, especially in areas where T. vulpis Vulpis and Ancylostoma Caninum Eggs infections are common.

Dr. Michael Dryden1, Dr. Brian Herrin1, Dr. Patricia Payne1, Vicki Smith1, Debra Ritchie1, OA10.07 US Dog Owner Compliance With Evan Hegarty2 Veterinary Recommendations for Heartworm 1Kansas State University, Manhattan, United Medication Use in Dogs That Also Receive States, 2IDEXX Laboratories, Westbrook, Flea and Tick Medication United States Dr. Robert Lavan1, Dr. Kathy Heaney1, Dr. Between 2010 and 2018, 239 individuals in 29 Srinivasan Rajagopalan2, Dr. Kaan Tunceli3 classes participated in a week-long clinical 1Merck Animal Health, Perkasie, United States, parasitology training program. Participants 2Med Data Analytics, New York City, United were either veterinarians (233) or persons States, 3Merck, Inc., Kenilworth, United States with advanced training in parasitology (6). As part of the course, attendees participated in a Many flea and tick medications, including fecal diagnostic wet lab. Fecal samples were some of the newer isoxazoline products, are collected from dogs at the local animal shelter given to dogs monthly. Many heartworm and verified as positive for various parasite prophylaxis medications are also re-dosed diagnostic stages. While species of parasites monthly. If dog owners use an extended and egg counts varied between classes, all duration flea and tick product, dosed at classes evaluated samples that contained 12-week intervals, then will they also be as Ancylostoma caninum, Toxocara canis compliant with veterinary monthly heartworm and Trichuris vulpis eggs. Each participant medication recommendations as owners conducted a direct smear, a commercial of dogs receiving a monthly flea and tick passive floatation device (Ovassay®) with product. 1.18 sp. gr. ZnSO4 solution, a centrifugation procedure using 1.18 sp. gr. ZnSO4 solution Clinic transaction records for over 200,000 and a centrifugation procedure using 1.27 dogs from approximately 650 veterinary sp. gr. sugar solution. Participants recovered clinics in the United States were examined A. caninum, T. canis, and T. vulpis eggs from for June, 2014 through November, 2017. Dog 71.9% (172/239), 61.1% (146/239), and 37.7% owners were identified as “pure users” and (90/239) of the samples, respectively when put in non-overlapping groups for particular using the Ovassay® device. When comparing flea and tick medications, meaning that they centrifugation techniques, participants were had purchased at least one dose of a flea/ more likely to recover T. vulpis eggs using the tick medication without switching to another higher sp. gr. sugar solution, 96.7% (231/239), product during a 12-month period. Brands of than using ZnSO4, 80.3% (192/239) (p<0.001). monthly flea/tick products were compiled as Recovery of A. caninum and T. canis did not a single comparator group. significantly differ between the centrifugation methods (pAC=0.81; pTC=0.25). Reliability The number of doses and treatment duration of the Ovassay® technique was significantly for purchases of 14 pooled heartworm inferior to centrifugation for each parasite medications were compared using two flea/ (pTCZ<0.001, pTCS<0.001, pACZ<0.001, tick prevention treatment groups (12-week pACS<0.001, pTVZ<0.001, pTVS<0.001). extended duration versus monthly duration). Trichuris vulpis eggs were recovered by every Dog owners who purchased a longer-acting participant in the class only 5% (1/29) of the flea and tick medication purchased slightly time by Ovassay® passive flotation, compared more heartworm medication annually for to 34.5% (10/29) and 75.8% (22/29) of the their dogs compared with dog owners who time by ZnSO4 or sugar centrifugation, purchase monthly flea and tick medication; respectively. This data provides further however, this increase is not large enough to evidence that passive fecal flotation is an be biomedically significant.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 49 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA11 Canine Protozoa 89 cardiac samples was positive for T. cruzi DNA. Fourteen (23%) of the 61 submitted July 8, 2019, 16:30 - 18:00 kissing bugs from Oklahoma were positive for Plenary Hall, Madison Ballroom (ABCD), Level 4 T. cruzi DNA; vertebrate host remnant blood meal identities could not be determined for the majority of dissected bugs, but seven OA11.01 Prevalence of Trypanosoma Cruzi (11.5%) individual kissing bugs were found Infection in Domestic Dogs and Kissing Bugs positive for human DNA, and one (1.6%) bug in Oklahoma was positive for canine DNA. These serologic and molecular data collected from domestic Miss Layna Tarpalechee1, Miss Megan dogs together with molecular data from Wohltjen1, Mrs Alexa Hunter1, Dr Susan Little1, volunteer submitted kissing bugs suggest that Dr Jerry Ritchey1, Mrs Jana Slaughter2, Dr T. cruzi may be an under recognized pathogen Justin Talley2, Dr Kelly Allen1 cycling in eastern Oklahoma. 1Department of Veterinary Pathobiology, Oklahoma State University Center for Veterinary Health Sciences, Stillwater, United OA11.02 Black-Backed Jackals and African States, 2Department of Entomology and Wild Dogs Serve As Reservoirs for the Plant Pathology, Oklahoma State University, Pathogenic Babesia of Domestic Dogs in Stillwater, United States South Africa

Trypanosoma cruzi is the triatomine-borne Ms Angy Shabangu1, Prof Banie Penzhorn1, hemoprotozoan causing Chagas disease Prof Marinda Oosthuizen1, Prof Paul Matjila1 in canines and humans in the Americas. 1Vector and Vector-Borne Research Clinical manifestations of the disease range Programme, Department of Veterinary from asymptomatic to acute myocarditis Tropical Diseases, Faculty of Veterinary and sudden death to chronic disease with Science, University of Pretoria, Pretoria, South eventual heart failure. Antibody screening Africa of owned or impounded dogs (n = 304) in eastern Oklahoma between years 1996–1997 Canine babesiosis caused by Babesia rossi is indicated an overall seroprevalence of 3.6%. a highly pathogenic disease of susceptible Recent surveys estimating T. cruzi infection domestic dogs. Babesia rossi occurs freely in prevalence in domestic dog populations indigenous wild canids and in domestic dogs within the state are lacking. In 2018, we in South Africa. The aim of this study was tested shelter dogs (n = 170) in eastern to investigate the occurrence of B. rossi in Oklahoma (Tulsa County and Le Flore or asymptomatic indigenous wild canids (black- surrounding county) for T. cruzi reactive backed jackals and African wild dogs) and in antibodies by ELISA (Chagas STAT-PAK susceptible domestic dogs. Assay®) and FFPE cardiac tissues from client- owned dogs diagnosed with myocarditis Wild canids blood samples were collected (n=89) between 2007–2017 at the Oklahoma from (an apparently) healthy populations Animal Disease Diagnostic Laboratory of free ranging black-backed jackals (n=75) (OADDL, Stillwater, OK) for molecular and captive black-backed jackals (n=25). evidence of T. cruzi microsatellite DNA. In Additional samples from free-ranging addition, we analyzed submitted kissing bugs African wild dogs (n=52) were also obtained, (n = 61) from 15 counties in Oklahoma for including 5 samples collected at a zoo from evidence of T. cruzi DNA in insect abdominal dead wild dogs. Domestic dogs samples extracts, as well as for the molecular (n=75) were collected from dogs suspected identification of remnant vertebrate blood with canine babesiosis. meal source. Thirty (17.5%) of the 170 canine samples were positive for T. cruzi reactive Reverse line blot hybridization assay revealed antibodies, which is a significantly higher occurrence of B. rossi in 29% of free ranging seroprevalence than previously documented black-backed jackals, 28% in captive black- in Oklahoma (p<0.0001), and one (1.1%) of backed jackals and 10% in free ranging African

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 50 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

wild dogs respectively. A majority of domestic In vitro studies: Attachment of Ap infected dogs (88%) were found to be positive for B. rossi. ticks in artificial seeding chambers was assessed and blood pools were sampled using Phylogenetic analysis revealed sequence qPCR at 6 hour intervals, up to 72h after first identity to B. rossi, based on the 18S rRNA female tick attachment. gene sequences from indigenous wild canids and susceptible domestic dogs. The B. rossi In vitro study 1: Sixty artificial feeding that occurs in asymptomatic indigenous wild chambers were seeded with 5 male and canids appears to be the same as the one that 5 female I. ricinus ticks each. Attachment occurs in susceptible domestic dogs. Thus, ranged from 20-60%. Ap DNA was detected this study was able to indicate that since in 5% of chambers already at 6h. The highest wild canids and domestic dogs share similar percentage of positive samples (16.3%) was pathogenic strain of B. rossi, wild canids may observed at 36h. serve as reservoirs of B. rossi. This highlights the importance of tick control, especially in In vitro study 2: Artificial feeding chambers domestic dogs as they are more susceptible (47 for I. ricinus, 34 for I. scapularis) were to B. rossi infections which appear to be less seeded with one adult male and female tick problematic to wild canids. each. Ap DNA was detected in 38.24% for I. scapularis and 4.26% for I. ricinus chambers at 6h. The highest percentage of positive OA11.03 Transmission of Anaplasma samples was observed for I. ricinus at 18h Phagocytophilum (Foggie 1949, Dumler et al. (21.95%) and I. scapularis at 24h (65.38%). 2001) by Ixodes Spp. Ticks Feeding on Dogs Conclusion: Transmission of Ap starts within and Artificial Membranes a few hours of tick attachment (within 6h in vitro). However detectable infections of Ap in Dr Josephus Fourie1, Mr Alec Evans2, Mr Riaan dogs are apparently dependent on a minimum Maree1, Dr Michel Labuschagne3, Dr Maxime inoculation dose. Madder3, Dr Matthias Pollmeier4, Dr Bettina Schunack4 1Clinvet International (Pty) Ltd, Bloemfontein, OA11.04 Prevalence of Vector-Transmited South Africa, 2Clinvet SA Morocco, Infections in Dogs in Berlin/Brandenburg Mohammedia, Morocco, 3Clinglobal, Black River, Mauritius, 4Bayer Animal Health GMBH, Christina Helm1, PD Dr. Jürgen Krücken1, Dr. Monheim, Germany Jana Liesner1, Prof. Dr. Manuela Schnyder2, Dr. Corinna Weber3, Dr. Elisabeth Müller3, The transmission time of Anaplasma Dr. Roland Schaper4, Dr. Stefan Pachnicke4, phagocytophilum (Ap) was evaluated in vivo Dr. Hans-Frieder Matthes5, Dr. Daniel using Ixodes ricinus ticks on dogs, and in Schaarschmidt-Kiener6, Prof. Dr. Peter vitro using I. ricinus and Ixodes scapularis on Deplazes2, Prof. Dr. Barbara Kohn7, Prof. Dr. artificial membranes. Infectivity of ticks was Georg von Samson-Himmelstjerna1 confirmed by qPCR on pools and ranged from 1Institute of Parasitology and Tropical 21 to 100%. Veterinary Medicine, Freie Universität Berlin, Berlin, Germany, 2Institute of Parasitology of In vivo study 1: Six groups (G) of 3 dogs each the University of Zurich, Zurich, Switzerland, were infested on Day 0 with Ap infected adult 3Laboklin GmbH & Co. KG, Bad Kissingen, ticks (50 in G1 to G5, 60 in G6). Ticks were Germany, 4Bayer Animal Health GmbH, either removed at 3, 6, 12, 24 and 48 hours Leverkusen, Germany, 5AFOSA-Animal (h) post-infestation (G1 to G5), or left until Welfare & Food Safety GmbH, Blankenfelde- engorged (G6). Starting Day 0 blood was Mahlow, Germany, 6Labor am Zuggersee, collected weekly for qPCR and serological Hünenburg, Switzerland, 7Clinic for Small analysis. Ap specific antibodies and DNA were Animals, Freie Universität Berlin, Berlin, detected in the 3 dogs in G6 only. No clinical Germany symptoms of Ap were observed. Vector-borne diseases are spreading because

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 51 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

of global warming, changing land use and OA11.05 Prevalence of Babesia Microti, socioeconomy. They pose important animal B. Odocoilei and Co-Infection Rates With and human health risks. A previous PCR- Borrelia Burgdorferi in Ixodes Scapularis based study had revealed low prevalences Nymphs in Wisconsin of vector-borne pathogens in dogs and no evidence for autochthonous transmission of Tela Zembsch1, Dr. Xia Lee1, Dr. Gebbiena Babesia spp., Ehrlichia canis and filaroids. Bron1, Dr. Lyric Bartholomay1, Dr. Susan Paskewitz1 Between May 2016 and April 2018, 1008 serum 1University of Wisconsin - Madison, Madison, samples of dogs from Berlin/Brandenburg United States were tested for presence of antibodies against or circulating antigens of vector- Babesiosis is a malaria-like disease caused by borne pathogens. Samples were tested parasites within the Babesia genus. Babesia with IDEXX SNAP®4Dx®Plus for antibodies microti, shares a mammalian reservoir, against Anaplasma spp., Ehrlichia spp. and Peromyscus leucopus, and a vector, Ixodes Borrelia spp. and Dirofilaria immitis antigen. scapularis, with the Lyme disease spirochete, Conventional ELISAs were used for detection Borrelia burgdorferi. Similarly, Babesia of antibodies against Anaplasma spp., Babesia odocoilei is transmitted by I. scapularis spp., Leishmania spp., Dirofilaria spp. and and infects white-tailed deer, Odocoileus Angiostrongylus vasorum and A. vasorum virginianus. The interactions of these disease antigen. Additionally, DNA samples from agents may allow for either competition or the previous study from dogs and foxes in amplification in tick or animal hosts. The two Brandenburg were screened for Hepatozoon main goals of this study were 1) to calculate canis by PCR. the prevalence of B. microti and B. odocoilei in I. scapularis nymphs from multiple locations While antibodies against Dirofilaria spp. were distributed across Wisconsin and 2) to detected in 22/1008 (2.2%) of the serum determine if there is an observable association samples, only 3 (0.03%) were positive for D. between these agents in the vector. Over immitis antigen. Furthermore, 66 (6.5%), 9 2,500 nymphs were collected from parks (0.1%) and 17 (1.7%) of 1008 samples were and natural areas throughout the state from positive for Anaplasma spp., Ehrlichia spp. and 2016-2018 and tested for pathogens via real- Borrelia spp. in the SNAP test. In contrast, the time PCR. Prevalence was calculated for the complete antigen-based Anaplasma ELISA individual pathogens and a Fisher’s exact detected 306/1001 samples (30.6%) positive. test was used to determine if an association Regarding A. vasorum, 29/1008 (2.9%) between either of the Babesia spp. and B. samples were antigen-positive, 35 (3.5%) were burgdorferi occurred. Overall, there was antibody-positive and 12 (1.2%) were double a strong tendency for ticks infected with positive. Leishmania spp. and Babesia spp. Babesia microti, but not B. odocoilei, to be antibodies were detected in 14/991 (1.4%) co-infected with Borrelia burgdorferi. Some and 26/1008 (2.6%) samples, respectively. ticks were co-infected with both Babesia Using the H. canis PCR, 46/1050 dogs (4.4%) microti and B. odocoilei, suggesting that the and 156/201 foxes (77.6%) were positive. latter parasite is transovarially transmitted Genotyping and travel history of dogs from female ticks to their eggs and offspring. suggest autochthonous transmission of H. Understanding the prevalence and biotic canis between foxes and dogs in the absence interactions between pathogens enhances of known vectors. In contrast, autochthonous veterinarians’ and wildlife managers’ ability to transmission is so far not a major issue for the recognize and respond to disease. Mediterranean and tropical parasite species in local dog populations despite of presence of vectors for some of the pathogens.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 52 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA12 Diagnosis and Decision Support and agreement coefficients were found for GI Nematodes in Ruminants II between mean FEC of individual and pooled samples. Values were in the same range for July 8, 2019, 16:30 - 18:00 the different pools (0.95 to 0.98 for rs and Breakout Room 2, Hall of Ideas E&H, Level 4 0.97 to 0.99 for c) and indicated that any of pooling strategy was efficient. In contrast, %FECR calculatedρ from individual FECs and OA12.01 Rapid Assessment of Faecal Egg from pooled FECs showed lower values for Count and Faecal Egg Count Reduction Spearman rs (0.67 to 0.80) and Lin’s c (0.49 Using Composite Sampling in Cattle to 0.74) meaning that composite sampling has a lack of reliability for estimating FECR%.ρ Dr Laura Rinaldi1, Alessandra Amadesi1, Dr Elaudy Dufourd2, Antonio Bosco1, Marion Gadanho2, Anne Lehebel2, Dr Maria Paola OA12.02 Using Remote Sensing Technologies Maurelli1, Dr Alain Chauvin2, Dr Johannes for the Detection of Parasitism in Sheep Charlier3, Dr Giuseppe Cringoli1, Dr Nadine Ravinet2, Dr Christophe Chartier2 Dr Seer Ikurior1, Professor Bill Pomroy1, Dr Ian 1University of Napoli Federico II, Napoli, Italy, Scott1, Dr Rene Corner-Thomas1, Dr Stephan 2Bioepar, Inra, Oniris, Nantes, France, 3Kreavet, Leu2, Dr Nelly Marquetoux1 Kruibeke, Belgium 1Massey University, Palmerston North, New Zealand, 2Macquarie University, Sydney, Faecal egg counts (FEC) and FEC reduction Australia (FECR) tests for assessing gastrointestinal nematode infection and anthelmintic Animals suffering from parasitism typically resistance are rarely carried out on ruminant display altered grazing behaviour and a farms because of the cost of individual voluntary reduction in feed intake. These analyses. The use of composite (pooled) changes are potentially important as faecal samples in which equal amounts of indicators of disease. Recent advances in GPS faeces from several animals are mixed and and accelerometer technologies provide the analyzed together is a promising method opportunity to objectively measure animal to reduce time and costs, but few studies behaviour while on pasture. GPS tracking are available, especially with regard to collars are suited to monitor and detect the evaluation of different pool sizes and changes in movement patterns. Triaxial its application in FECR test. A total of 29 accelerometers measure body movement groups of cattle were investigated in Italy in terms of acceleration, which can then and France (9 to 20 animals/group). In each be used to estimate physical activity over group, individual faecal samples from heifers time. Two preliminary studies using these (6 to 20 month old) were collected before technologies are presented. In study one, (D0) and two weeks after (D14) anthelmintic the accuracy and performance of seven treatment (ivermectin or benzimidazoles). commercially sourced GPS receivers were FECs were determined individually and as assessed. A motion test was used to assess pooled samples using the Mini-FLOTAC the receivers’ prediction of distance travelled technique. Three pool sizes were used from against the inner lane of an athletics track. all the animals of each group: pools of 5 or 10 Stationary tests assessed how dispersed individual samples or global pool. The mean location estimates from the receivers were FEC of individual and pooled samples were from a “true” location landmark. Six of seven calculated as the arithmetic mean and FECR% receivers returned <5 m errors in location was [1 − (arithmetic mean FEC post treatment estimates. The implication of error rates for ÷ arithmetic mean FEC pre- treatment)] x measuring distance travelled in parasite- 100%. Correlations and agreements between infected and uninfected animals is discussed. individual and pooled results (FEC and In study two, the total activity levels of two FECR%) were estimated with the Spearman’s groups of crossbred Romney and Suffolk ram and the Lin’s correlation coefficients lambs were monitored over a period of four respectively (rs and c). High correlation days using triaxial accelerometer sensors

ρ

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 53 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

after 42-days exposure to treatment. On were used. Three biological replicates were day zero, all lambs were given anthelmintics. performed for each parasite isolate using Subsequently, a Suppressive Treatment Group both eprinomectin and ivermectin. Eleven (S) (n=12) was treated with anthelmintics drug concentrations from 0.01um to 40um every two weeks. An Untreated Group (U) and negative controls were evaluated. Motility (n=12) did not receive anthelmintics. Total readings were taken using the Worminator activity levels were monitored from days’ 42 system before addition of the drug and at 24 – 46. Activity level was calculated as vectorial and 48 hours post drug exposure. Differences dynamic body acceleration (VeDBA). Parasite in EC50 for ivermectin were highly variable burdens measured by faecal egg counts were between susceptible and resistant isolates; low (mean S = 50 eggs/g; mean U = 500 resistance ratios (RR) ranged from 0.425- eggs/g). Activity levels in untreated lambs 9.47. In contrast, eprinomectin assays had were significantly less than in the Suppressive much less variability and consistently yielded Treatment group (P=0.024). This small but similar EC50 for all isolates (RR of 0.958-1.04). significant difference in total activity indicates Though significant differences (p<0.05) in usefulness of this approach in detecting percent inhibition were found at some drug subclinical parasitism. concentrations in some assays, there were no consistent significant differences in the dose- response across comparative assays or for the OA12.03 Motility Based Assays Using different drugs. Inhibition was greater for the Cultured Fourth Stage Larvae Fail to Provide susceptible isolate in about half of the assays. Consistent Discrimination Between Known The lack of consistency in these data indicates Avermectin-Resistant and -Susceptible that motility of L4 is not a reliable diagnostic Isolates of Cooperia spp.Motility Based phenotype for measuring resistance to Assays Using Cultured Fourth Stage Larvae avermectin drugs in Cooperia spp. Fail to Provide Consistent Discrimination Between Known Avermectin-Resistant and -Susceptible Isolates of Cooperia Spp. OA12.04 What Are Good Marker Genes for Strongyle Nematode Species Identification Kelsey Paras1, Ray Kaplan1 and Phylogenetic Reconstruction? 1University of Georgia, Athens, United States Sabrina Ramünke1, Christina Bredtmann1, The fecal egg count reduction test (FECRT) Tetiana Kuzmina2, Mariana Louro1,3, Fernando is the only method commonly used for de Almeida Borges3, Elke von Son-de Fernex4, diagnosing anthelmintic resistance in GIN of Luís M. Madeira de Carvalho3, Georg von cattle, but is time-consuming and potentially Samson-Himmelstjerna1, Jürgen Krücken1 cost-prohibitive. Consequently, there exists 1Freie Universität Berlin, Berlin, Germany, a need to develop better methods for 2National Academy of Sciences of Ukraine, diagnosing resistance. Assays based on larval Kiev, Ukraine, 3University of Lisbon, Lisbon, motility are used commonly for screening Portugal, 4Universidade Federal do Mato potential drug candidates, but previous work Grosso do Sul, Campo Grande, Brazil, in our lab demonstrated that the L3 stage 5Universidad Nacional Autonoma de Mexico, failed to discriminate between avermectin- Veracruz, Mexico resistant and susceptible isolates of Cooperia spp. The L4 may be a better stage for this Species descriptions and identification purpose because it is a parasitic and actively keys to discriminate closely related feeding life stage without a double cuticle. strongyle nematodes rely on differences The L4 were cultured by exsheathing L3, in morphological traits – particularly of maintaining them in media at 37°C and 20% the buccal capsule and the male bursa. CO2, with media changes and observation In the absence of extensive data on intra- every 48 hours for nine days. Three species morphometric variation, molecular avermectin-resistant and two avermectin- approaches have mainly focused on the susceptible GIN isolates (diagnosed by intergenic-spacer (ITS) of the nuclear FECRT) containing >90% Cooperia spp., ribosomal genes, the mitochondrial genome,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 54 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

particularly the cytochrome oxidase I (COI) Classic approaches for identifying and the 12S and 16S rRNA genes. Using data gastrointestinal nematodes (GIN) from on Cooperia species infecting cattle and on fecal eggs require cultivation to infective equine Cyathostominae, this study analysed L3 followed by morphological examination. the suitability of different nuclear and Over the years, a plethora of molecular mitochondrial sequences for reliable species techniques have surfaced, most which are identification and phylogenetic reconstruction predicated upon real-time PCR or PCR using maximum likelihood. Using ITS-2 amplification followed by gel analysis. While sequences, most but not all species could be adequate for diagnosis, many techniques fall differentially identified using phylogenetic short of easily quantifying mixed infections analyses but not by simple comparison of without substantial numbers of controls and/ identities. In contrast, mitochondrial markers or standard curves. Herein we developed a had much better barcoding properties, but simple and rapid test for differentiating and resulted in poor phylogenetic resolution, both quantifying mixed infections of GI nematodes due to saturated variation in codon position using fluorescently-labeled PCR products and 3. Optimal trees were obtained by partitioned a capillary-based sequencer. The assay can analyses of both nuclear and mitochondrial be easily scaled for large scale analyses and markers. In Cooperia, multi-locus analysis of does not require external controls. Among sequences obtained from individual worms the common cattle GIN, the ITS2 region is clearly discriminated Cooperia oncophora sufficiently distinct in length to delineate and Cooperia pectinate. However, Cooperia among infecting genera. As such, conserved punctata and Cooperia spatulata always PCR primers that span the ITS2 and bind to formed a common cluster, suggesting that the the 3’-end of the 5.8S rRNA (forward) and latter is only a morphotype of C. punctata. the 5’-end of the lsurRNA (reverse) were In the Cyathostominae, close relationship synthesized, one of which was fluorescently- between Coronocyclus coronatus and labeled with FAM. DNA from infective L3 Cylicostephanus calicatus was revealed by was isolated, PCR amplified, diluted directly virtually identical ITS-2 sequences. Concurrent in HI-DYE sequencing buffer containing analysis of nuclear and mitochondrial LIZ 500 standard, then loaded onto an ABI sequences allowed the discrimination 3100 sequencer adapted for size fragment between mitochondrial lineage sorting and analysis. Validation was first performed on potential speciation. Cryptic species were monospecific infections of Haemonchus, detected in Cylicostephanus minutus (3), Ostertagia, Cooperia, Trichostrongylus and Cys. calicatus (2) and potentially Cylicocyclus Oesophagostomum. As proof of principle, nassatus (3). Cylicostephanus labiatus L3 from Ostertagia ostertagi, Cooperia showed diverse mitochondrial and nuclear punctata and Haemonchus contortus were haplotypes that were mixing with each mixed in pairs and in 10% increments, other, while Cylicostephanus longibursatus followed by DNA isolation and analysis for was comparatively homogenous. In relative DNA levels using Gene Marker V1.85. conclusion, combined analysis of nuclear In all cases, experimental data generated and mitochondrial haplotypes from the a linear response and correlated well with same morphologically identified specimens, predicted values where R2> 0.95 with line improved resolution of analyses and should be slopes between 0.90-1.1. The assay was tested applied to more species and specimens from on environmentally-derived samples and various geographic regions. coincided well with coproculture, PCR gel analysis and deep amplicon sequencing. Data show that primer design and purification OA12.05 A Simple Method for Identifying are critical to eliminating satellite bands and and Quantifying Cattle GI Nematodes enhancing quantitative analyses.

Dr. Dante Zarlenga1, Dr. Carly Barone1, Ms. Deborah Hebert1 1Agricultural Research Service, Animal Parasitic Diseases Lab, Beltsville, United States

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 55 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA12.06 Investigating the Diversity and OA13 Flea and Tick Treatment Prevalence of Pathogens in Cattle in the Livestock-Wildlife Interface of Kwale County, July 8, 2019, 16:30 - 18:00 Kenya. Breakout Room 3, Hall of Ideas F&I, Level 4

Brenda Odhiambo1, Dr Shewit Kalayou1, Dr Dan Masiga1, Dr Michael Okal1 OA13.01 Efficacy of Oral Afoxolaner 1International Centre For Insect Physiology (NexGard®) Against Cochliomyia Hominivorax And Ecology, Nairobi, Kenya (Diptera, Calliphoridae) Larvae Infestation in Naturally Infested Dogs in Brazil Livestock keepers in areas bordering wildlife protected areas in Africa bear the brunt of Andre Cutolo1, Ingrid Menz2, Patricia Thyssen3, bovine trypanosomosis and a plethora of Fabiano Silva4, Fred Beugnet5 emerging and re-emerging pathogens. In 1Boehringer Ingelheim Animal Health Do Kwale, Kenya, bovine trypanosomosis has Brasil Ltda., Paulínia, Brazil, 2Ingrid Menz ME, been the focus of many interventions around Campinas, Brazil, 3Universidade Estadual the protected wildlife hotspot of the Shimba de Campinas - Unicamp, Campinas, Brazil, Hills National Reserve (SHNR). Though some 4Boehringer Ingelheim Animal Health, Fulton, studies have detected tick-borne pathogens United States, 5Boehringer Ingelheim Animal in or near the SNHR, the prevalence and Health, Lyon, France overlap of these remains unknown. This study implemented a longitudinal survey in four New World screwworm disease is an villages around the Shimba Hills National infestation with the larvae of the screwworm Reserve in Kwale to determine the diversity fly (Cochliomyia hominivorax) that lives off of pathogens in cattle. A cohorts of 120 cattle the flesh of living warm-blooded animals, from twelve households within 500 m from including humans. Domestic dogs are among the park were screened for bacterial, viral the potential hosts of the fly that lay eggs and protozoal pathogens. All households on pre-existing bleeding open wounds were from villages at least three kilometers where larvae feed and develop on live tissue. apart. Rapid and high-throughput screening Disease can be fatal if untreated. Treatment of the samples for pathogens was done using with parasiticides must be quick-acting, the high resolution melting analysis of PCR 100% efficacious and long-lasting, since products. About 30.8% of livestock were open wounds can be reinfested. Afoxolaner infected with Trypanosoma congolense. 16.7% is a molecule from the isoxazoline family of animals were infected with one of four with proven ectoparasiticide action against Anaplasma sp (An, ovis, An. marginale, An. fleas and ticks infesting dogs. Afoxolaner platys or An. phagocytophilum, 15.5% had (NexGard®) was administered from doses Theileria velifera, 13.5% Erhlichia sp. and 9.7% ranging from 2.7 mg/kg to 6.7 mg/kg Rickettsia sp. Viral infections were detected (same dose as to control fleas and ticks) on in 37.3% of the sampled livestock. These were dogs naturally infested with Cochliomyia predominantly bunya (38.9%), thogoto viruses hominivorax larvae in the municipality of (40.0%) and phlebo (12.2%) viruses, with a Campinas, São Paulo state, Brazil. From March few nairo (5.6%) and alpha (3.3%) viruses also 2017 to April 2018, 14 dogs having active detected. These results highlight the complex lesions infested with C. hominivorax larvae disease environment of the wildlife-livestock were included in the study. Infestations were interface in Kwale and, by extrapolation that classified according to the number of larvae of agro-pastoralism near wildlife protected recovered post-treatment as light (less than areas in Kenya. The collective potential impact 10 larvae), moderate (from 10 to 20 larvae) of these on the productivity and production and severe (more than 20 larvae). No adverse of cattle and their impact on the success of events were observed after treatment. Two controlling of bovine trypanosomosis, though hours after treatment intensification of larval not investigated here is discussed. movements on the lesions was observed. Six hours post-treatment it was possible to observe that some larvae were expulsed from

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 56 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

the lesions. Twenty four hours after treatment synthesis of the cRNAs, which were injected all larvae were killed on all treated animals. in the Xenopus oocysts system to reconstitute Total amount of larvae recovered ranged from functional channels. Two-electrode voltage 3 to 327 larvae per animal (average of 67.7). clamp recordings showed that Phh-RDL Afoxolaner (NexGard®) at doses ranging from assemble into a homomeric receptor sensitive the minimum active dose of 2.5 mg/kg to the to different insecticides like fipronil, picrotoxin maximum dose 6.7 mg/kg (mean of 3.9 mg/ and lotilaner, a novel class of ectoparasiticide kg) was rapidly efficacious for the treatment agent using to treat ticks and fleas of dogs of 14 dogs with light, moderate and severe (CredelioTM, Elanco). These results correlated infestations with Cochliomyia hominivorax with the efficacy of these drugs on lice in vivo. larvae, by eliminating all larvae within 24 In conclusion, we report the functional hours after a single oral treatment. characterization of the first GABACl of Pediculus humanus humanus. These results contribute to our understanding of the mode OA13.02 Lotilaner Is a Potent Inhibitor of of action of insecticide compounds and will the Novel GABA Receptor of Body Lice allow the development of new therapeutic Pediculus Humanus Humanus strategies to control lice infestations.

Nicolas Lamassiaude1, Dr Berthine Toubate2, Dr Pierre Charnet3, Dr Cédric Neveu1, Dr OA13.03 Early Oral Sarolaner (SimparicaTM, Françoise Debièrre-Grockiego2, Dr Claude Zoetis) Acaricidal Activity Against Ixodes Charvet1, Pr Isabelle Dimier-Poisson2 Scapularis and Amblyomma Americanum 1ISP, INRA, Université Tours, UMR1282, Adults After Monitored Attachment Intervals Nouzilly, France, 2ISP, UFR de Pharmacie, on Treated Dogs Université Tours, UMR1282, Tours, France, 3Institut des Biomolécules Max Mousseron Dr. Kelly Allen1, Dr. Ruth Scimeca1, Megan (IBMM), CNRS UMR 5247, Montpellier, France Wohltjen1, Dr. Mason Reichard1, Nadia Koziar2, Deborah Amodie2, Dr. Georgette Wilson2, Dr. Drug resistance in the parasites field, including Joyce Login2 the cosmopolitan lice (Pediculus humanus), 1Oklahoma State University, Stillwater, United and the prevalence increasing despite the States, 2Zoetis Inc., Parsippany, United States marketing of new therapies are an important challenge for our societies. The major Acaricidal activity of oral sarolaner pharmacological targets of insecticides like (SimparicaTM, Zoetis) against Rhipicephalus pyrethrins, malathion, spinosad and ivermectin sanguineus sensu lato and Dermacentor (also used as nematicide and acaricide) are reticulatus begins 4 and 8 hours, respectively, the ligand gated ion channels present in the after induced infestations on treated dogs. nervous system of insects. Currently, targets Here, we evaluated Ixodes scapularis and of these molecules remain largely unknown in Amblyomma americanum mortality in body lice. Among those channels receptors, response to sarolaner after monitored, γ-aminobutyric acid gated chloride ion documented attachment time intervals channels (GABACl) are the main synaptic within 1–8 hours of infestation. In duplicate, inhibitory receptors in insects, making them staggered infestation experiments, 6 treated pertinent pharmacological targets. and 6 control dogs were infested with 250 I. scapularis and 250 A. americanum In the present study, we identified and adults. Ticks were allowed 60 minutes to characterized the targets of insecticides in lice embed mouthparts into the dermis. A ≤ 50% to decipher the mode of action of insecticides attachment rate was targeted. Subsets of in Pediculidae. Research in the genomic ≤ 20 ticks were removed from dogs after databases of Pediculus humanus allowed us documented attachment times of 1, 3, 5, and 7 to identify a GABACl subunit encoded by the hours or 2, 4, 6, and 8 hours. Live/dead status Resistance to dieldrin (Rdl) gene. We cloned was assessed at the time of removal and 24 the corresponding full-length cDNA into a hours post-removal (live ticks incubated at transcription vector and performed in vitro ambient temperature (~70°F [21.1°C], 80–90%

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 57 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

humidity). Geometric means of combined live present study, we characterized the GABACls tick counts were calculated for attachment from the European tick species Ixodes ricinus. intervals of 1–2 hours, 3–4 hours, 5–6 hours, and 7–8 hours, and least squares means We extracted RNAs from Ixodes ricinus were compared by ANOVA with a two-sided nymphs. Taking advantage of the significance level set at =0.05. Significantly phylogenetic closeness of I. ricinus and R. fewer live I. scapularis were removed from microplus in the Arthropoda phylum, we treated than control dogsα after 3–4 hours identified the I. ricinus GABACl subunit of attachment (p=0.0012); upon holding 24 homologue. The cDNA encoding the Iri- hours, significantly fewer I. scapularis that had GABACl was cloned and the corresponding in previously been attached to treated dogs for vitro synthesized cRNAs were micro-injected 1–2 hours were alive (p=0.0049). Significantly into Xenopus laevis oocytes to investigate fewer live A. americanum were removed from its pharmacological properties. Functional treated than control dogs after 7–8 hours expression and two-electrode voltage clamp of attachment (p=0.0254); upon holding studies demonstrated that the GABACl 24 hours, significantly fewer A. americanum subunit formed a homomeric receptor gated that had previously been attached to treated by GABA. Importantly, the insecticides like dogs for 3–4 hours were alive (p=0.0003). lotilaner, fipronil and picrotoxin efficiently These data indicate that acaricidal activity blocked the GABA currents as previously of sarolaner against I. scapularis and A. observed for the R. microplus GABACl. americanum begins after 1–2 hours and Surprisingly, I. ricinus GABACl was not 3–4 hours of attachment, respectively, on sensitive to the pesticide dieldrin, suggesting sarolaner treated dogs. a potential naturally existing resistance mechanism involving alternative exons.

OA13.04 A First Insight Into the Effect of Here we report the functional characterization Lotilaner on GABA-Gated Channels From the of the first GABACl of I. ricinus demonstrating European Tick Ixodes Ricinus that it is an important molecular target for lotilaner. Transcriptomic analysis of I. ricinus are Dr Claude Charvet1, Clément Auger1, Dr in progress to identify new acaricidal targets. Claude Rispe2, Caroline Hervet2, Dr Elise Courtot1, Pr Steeve Thany3, Dr Olivier Plantard2, Dr Cédric Neveu1 OA13.05 Clinical Efficacy of Afoxolaner in 1INRA, ISP, Université Tours, UMR1282, Dogs Naturally Infected With Sarcoptes Nouzilly, France, 2INRA, Oniris, BIOEPAR, Scabiei and Concomitant Modifications of Nantes, France, 3Université d’Orléans, LBLGC, the Skin Microbiota USC1328 INRA, Orléans, France Raluca Mindru2, Andrei Cristian Lupu2, Dr. Ticks are strict blood-feeding arthropods Constantin Roman2, Dr. Charlotte Bernigaud5, (Acari), which represent a major health issue Dr. Amaury Briand1, Dr. Frédéric Beugnet3, for wild or domesticated animals and humans, Dr. Katja Fischer4, Professor Liviu Miron2, due to their potential to transmit disease Professor Jacques Guillot1 agents. Control of ticks is increasingly difficult 1Veterinary College of Alfort, Maisons-alfort, due to the development of drug-resistant France, 2Ion Ionescu de la Brad, Faculty parasites. Ligand-gated ion channels of the of Veterinary Medicine, Iasi, Romania, tick central nervous system are the primary 3Boehringer Ingelheim Animal Health, Lyon, targets of acaricides. Among those receptors, France, 4QIMR Berghofer, Medical Research the γ-aminobutyric acid-gated chloride ion Institute, Brisbane, Australia, 5Henri Mondor channels (GABACls) are the main synaptic Hospital, France inhibitory receptors. Lotilaner is a recently developed parasiticide from the isoxazoline Infection by Sarcoptes scabiei remains chemical class that was shown to be a non- a common disease in dogs, especially in competitive antagonist of GABACls from the tropical countries where most of stray livestock tick Rhipicephalus microplus. In the or non-controlled dogs show cutaneous

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 58 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

lesions evocative of chronic infection. In flea. They are responsible for irritating pets Europe, cases are also regularly reported in and their owners as their bites can induce shelters or in privately-owned dog facilities. pruritus and allergic reactions. They are also Isoxazolines are insecticides and acaricides responsible for the transmission of pathogens of a new chemical class. The objectives of the of zoonotic and veterinary importance (i.e. present study were to evaluate the efficacy of Bartonella sp, Rickettsia sp, Dipylidium afoxolaner in twelve dogs naturally infected caninum …). Ectoparasiticide products by S. scabiei in Romania and to describe the represent the mainstay for preventing and modifications of the skin microbiota during halting insect bites and transmission of the acaricide treatment. We performed an arthropod-borne infections in companion open pre-treatment versus post-treatment animals. Since the late 80s, fipronil has been study enrolling dogs with clinical signs of one of the most used insecticides on pets sarcoptic mange and a positive skin scraping worldwide. In the past 10 years, veterinarians examination. Dogs were mixed-bred, ranging reported a lack of efficacy when using this between 4 months and 7 years old, and molecule for flea control. So far, the resistance weighing between 2 and 20 kg. They received of fleas to insecticide has not been properly oral afoxolaner (2.7-6.9 mg/kg) on days 0 studied. and 28. Treatment efficacy was determined by the reduction of a clinical score based on The aim of our work was to investigate the the skin surface affected by scabies lesions, mechanisms underlying resistance to fipronil erythema intensity, and the presence of in a laboratory flea strain originated from a scales and crusts. The score was calculated field population resistant to fipronil and to on day 0 and subsequently until day 28 for 4 compare to a reference susceptible strain. anatomic sites (head, trunk, legs, tail). For 3 The phenotypic resistance was investigated random animals, skin scrapings were collected via tarsal contact using filter papers on two sites (ear and hind leg) before (D0) impregnated with increasing doses of fipronil and after (D28) treatments to assess the skin and dieldrin. The resistant strain showed a microbiota. DNA was extracted using DNA resistance ratio at LD50 of 6.03 and 33.23 for Midi Kit QiaSymphony (Qiagen) and analyzed fipronil and dieldrin respectively. The activity by bacterial 16S rDNA (V3-V4) and fungi ITS1 of detoxification enzymes was investigated amplicon sequencing (MiSeq Illumina). On day in adult fleas and third-stage larvae. The 0, mean clinical scores were 24.7 (4-58). On results showed a significant increase in day 28, all dogs were mite-free and clinical the activity of the and esterases in the scores declined to 9.5% (1-28). No signs of adults and larvae of the resistant strain. A drug intolerance were noticed. We found significant decreaseα in theβ cytochrome P450 changes in the skin microbiota after the initial mono-oxygenases in the resistant larvae acaricide treatment. was also observed. Phenotypic assays with detoxification enzyme inhibitors are underway to confirm these results. The molecular assays OA13.06 Ctenocephalides Felis Resistance showed that Rdl mutation is established in to Fipronil: Phenotypic and Genotypic resistant fleas. Monitoring these mechanisms Characterization in a Laboratory Model in natural populations could greatly improve insecticide resistance management and help Dr Emilie Bouhsira1, Dr Emmanuel Liénard1, development of new formulations to control Mr Audric Berger2, Dr Shukri Sharif1, Pr flea infestations. Michel Franc1, Dr Fabrice Chandre2, Dr Sylvie Cornélie2 1National Veterinary School of Toulouse, Toulouse, France, 2Institute for Research and Development, Montpellier, France

Fleas are considered as one of the major pests of domestic animals, especially Ctenocephalides felis, also known as the cat

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 59 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA14 Protozoan Parasites attributed to relapsing EPM in these horses and should be considered in evaluating July 8, 2019, 16:30 - 18:00 neuromuscular disease. Breakout Room 4, Hall of Ideas G&J, Level 4

OA14.02 Bovine Theileriosis – A New Disease OA14.01 Defining Relapses Attributed to of Cattle in Australasia Equine Protozoal Myeloencephalitis Dr. Abdul Jabbar1 Dr. Siobhan Ellison1 1Melbourne Veterinary School, The University 1Pathogenes Inc, Reddick, United States Of Melbourne, Werribee, Australia

The consensus opinion (Reed et al 2016) Bovine theileriosis is a tick-borne disease indicate that cycles of clinical improvement caused by protozoan parasites of the followed by relapse often seen in treated cases genus Theileria, particularly in subtropical of equine protozoal myeloencephalitis (EPM) parts of the world. In Australia, Theileria can be due to re-exposure to S. neurona (SN) was first detected in cattle in the early sporocysts or persistent latent but unapparent 1900s, and infection has become endemic infections. Persistent S. fayeri (SF) infections in some regions of the east of the country. were linked to clinical disease by Aleman et The infection is thought to have been al (2014). Recurring disease after treating introduced from Japan into Australia via cattle sarcocystosis in horses, post-treatment EPM infested with the ixodid tick Haemaphysalis disease syndrome (PTEDS), and co-infections longicornis. Traditionally, Theileria infection in with S. neurona/fayeri (SN/SF) may complicate cattle in Australia was usually asymptomatic evaluating neuropathies in horses. In this study and recognized to be associated with Theileria we investigated the relationship between buffeli, although some clinical cases had recurrent clinical neurological disease and been reported in Queensland in the 1960s. seroprevalence of species-specific antibodies T. buffeli is now recognized as one of the of equine-infecting Sarcocystis by prophylaxis members of the Theileria orientalis complex. using decoquinate to identify specific aspects Since 2006, there have been many outbreaks of disease progression. The possibility that of theileriosis in dairy and beef cattle, mainly clinical signs were due to inflammatory in New South Wales and Victoria, which have or immune mediated complications after been linked mainly to two ‘virulent’ genotypes resolution of active infections was examined ikeda and/ or chitose of this complex. A by the presence of circulating anti-myelin basic recent study showed that bovine theileriosis P2 (MPP) antibody. Sixty-nine clinically normal can lead to significant milk production losses horses with a history of chronic relapsing EPM in dairy cattle. Although Theileria spp. known were grouped by seroreactivity (SN, SF +SN, to infect cattle were present in Australia since or MPP +SN/SF) and treated daily with oral the early 20th century, it is believed that the decoquinate until clinical relapse was apparent. pathogenic genotypes of T. orientalis were The horses in the SN group did not relapse, recently introduced to the country. three horses in the SF group (10%) relapsed in three months, and nine horses in the MPP group (35%) relapsed in the first three months. OA14.03 Epidemiology of Theileria The study showed that most relapse cases Orientalis: Potential Vectors on Northern were found in horses seropositive for MPP Tablelands, NSW, Australia antibody and not re-exposure to Sarcocystis. Decoquinate prophylaxis was statistically Mr. Biniam Tsegaye Lakew1, Professor Nigel R. more effective (p < 0.05) for preventing Andrew1, Associate Professor Shubiao Wu1, Mr sarcocystosis than preventing polyneuritis Steve Eastwood2, Professor Stephen Walkden- equi defined by MPP antibody and clinical Brown1 signs. This study indicates that PTEDS may 1University Of New England, Armidale, be unrelated to active infection. Polyneuritis Australia, 2Local Land Services, Northern equi was the leading cause of clinical signs Tablelands, Armidale, Australia

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 60 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Theileriosis is an economically important tick- OA14.04 Evaluation of an ELISA Detecting borne disease of cattle in tropical and sub- S. Fayeri Antitoxin in Horses With Equine tropical countries with Theileria parva and Muscular Sarcocystosis Theileria annulata being the most pathogenic species. Although Theileria orientalis is Dr. Siobhan Ellison1, Dr. Laura Benedetti1, Ms known to cause persistent infections with Julie Osborne1, Dr. Steve Duniho2, Dr. Craig mild signs, in recent years there has been Reinemeyer3 an increase in severe clinical outbreaks 1Pathogenes Inc, Reddick, United States, including mortality in Asia-Pacific region. 2Talon Pathology Services, Bellingham, United In Australia, such outbreaks commenced in States, 3East Tennessee Clinical Research, 2006 with new genotypes Ikeda and Chitose Rockwood, United States apparently responsible for clinical disease despite ongoing widespread prevalence of Disease due to Sarcocystis fayeri is associated the nonpathogenic genotype Buffeli. In New with neuromuscular disease in horses and South Wales, outbreaks were initially seen in toxin-induced food poisoning in people. higher rainfall coastal areas with infection now Equine muscular sarcocysts are found on endemic. The situation in nearby inland areas postmortem exam and generally considered having lower tick populations is less clear incidental. We report the presence of although outbreaks do occur. In the Northern circulating S. fayeri antitoxin in a group of 32 Tablelands of NSW clinical outbreaks have clinically normal horses that had incidental occurred since 2013 even during cold winter findings of muscular sarcocysts. Sarcocystis months when tick activity is low. As part of a fayeri anti-toxin was present in horses with study on the abundance and infection status and without sarcocysts. The results of this of potential vectors for T. orientalis in this report show similar sensitivity for detecting S. region, 89 bovine blood samples from eight fayeri sarcocysts in horses by histopathology farms within 100 km from Armidale were and S. fayeri anti-toxin (78% and 74% subjected to qPCR assays to quantify and respectively). These results indicate that S. differentiate various T. orientalis genotypes. fayeri anti-toxin may be a useful premortem All samples were positive for T. orientalis with screening test to detect the presence of S. 84%, 91% and 88% positive for Ikeda, Chitose fayeri exposure in horses. and Buffeli respectively. With regard to co- infection with different genotypes 70% of samples were positive for 3 genotypes, 24% OA14.05 Not All Species Are Created Equal: for 2 genotypes and 7% for a single genotype. Exploring the Diversity of Eimeria Spp. Based on published clinical thresholds for Infecting Domestic Sheep and Goats parasite counts of the pathogenic Ikeda genotype 4/89 (4.5%), 28/89 (31.5%) and Evelin Rejman1, Dr. John R. Barta1 57/89 (64.0%) fell into the high, moderate and 1Department of Pathobiology, Ontario low categories respectively with significant Veterinary College, University of Guelph, variations between farms. The results indicate Guelph, Canada that a high prevalence of co-infection with both pathogenic and non-pathogenic Coccidiosis, caused by the protozoan genotypes suggestive of endemic status. parasites in the genus Eimeria, is a costly Investigations into potential biological and disease impacting the global small ruminant mechanical vectors for T. orientalis in this industry mainly through reduced animal region are ongoing. performance and cost of treatment. Both clinical and subclinical disease cause a decrease in animal performance through decreased feed efficiency and weight loss from gut damage, loss of electrolyte and nutrients from diarrhea and, in severe cases, death of young livestock. There are at least 12 Eimeria species that infect domestic sheep and 9 Eimeria species that infect domestic

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 61 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

goats but only a few are considered to be Anthelmintic resistance in equine highly pathogenic and cause severe clinical cyathostomin parasites is widespread. A disease. The severity of disease is assessed surveillance-based parasite control programs typically using counts of fecal oocysts (i.e. using fecal egg counts (FEC) and fecal egg oocysts per gram, OPG) obtained using a count reduction tests (FECRT) to decrease modified McMaster method. Less pathogenic anthelmintic use and monitor treatment species can be excreted at high OPG counts efficacy is recommended. Information is often but have minimal clinical impact whereas distributed through mass media, word of pathogenic species can cause damage mouth, and veterinarians, but the impact of with low OPG counts. Consequentially, educational campaigns such as local extension conventional enumeration methods may outreach programs are rarely studied. The be unreliable for assessing the severity purpose of this study was to examine shifts of infections. Traditional identification of equine parasite control program management Eimeria species using morphometrics of practices due to a short course presented by oocysts is unreliable due to overlapping the Penn State Extension, and to highlight measurements and few species-specific how data collected from these programs is morphological features. Eimeria species useful for monitoring anthelmintic efficacy can be identified using sequence based on a large scale. Horse owners were enrolled genotyping (eg. Next Generation Sequencing) after participating in a short course, and of mitochondrial CDS or nuclear 18S rDNA then filled out questionnaire surveys about loci. The accurate identification of the their parasite management programs pre causative Eimeria spp. is a critical step in and post study, horse information, and disease management. This project is focused farm information. FECs were performed at on using sequencing techniques and PCR with three time points, and horses above a 200 species specific primers to identify Eimeria strongyle eggs per gram cut-off were treated spp. infecting domestic small ruminants from with pyrantel, fenbendazole, or ivermectin. populations that range in complexity from Two weeks post-treatment a FECRT was individual farms to regional epidemiological performed to determine treatment efficacy, studies that include multi-farm sampling. which included 29 farms with a total of 513 Combining parasitological parameters, individual treatments. Prior to the study, only such as identification of Eimeria spp. using 30.6% of farms used FECs, but after the study sequencing or PCR techniques, with animal 97% of farms said they would use FECs in performance measures (e.g. animal weights) the future. Horses were given an average of can improve the ability to accurately diagnose 4.1 anthelmintic treatments per year before the severity of coccidial infections and assess the study, and post study 86% of farms were the efficacy of anticoccidial treatments. able to reduce the number of anthelmintic treatments used. Fenbendazole was effective on zero farms, pyrantel on 7.4% of farms, and OA15 Education ivermectin on 92.9% of farms. This extension outreach project helped generate information July 8, 2019, 16:30 - 18:00 about anthelmintic efficacy levels, causing a Breakout Room 5, Meeting Rooms KLOP, Level 4 shift in practices on participating farms, and collected useful anthelmintic resistance data. Programs such as this could be useful in other OA15.01 The Importance of Anthelmintic states for monitoring resistance and helping Efficacy Monitoring: Results of an Outreach shift management practices. Effort

Jennifer Cain1, Donna Foulk2, Ed Jedrzejewski2, Heather Stofanak2, Martin Nielsen1 1University Of Kentucky, Lexington, United States, 2The Pennsylvania State University, University Park, United States

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 62 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA15.02 Teaching Horse Parasitology in the own pace, supplementing course material 21st Century: An Adaptive Learning Platform with multiple pathways designed to engage critical thinking while supporting students Ms. Nichola Calvani1, Dr. Gaby Van Galen1, Dr. with different levels of knowledge. The result Kris Hughes2, Dr. Anne Beasley3, Dr. Shokoofeh is an equine parasitology diagnostic module Shamsi2, Dr. Jan Slapeta1 that students can repeat as necessary, while 1Sydney School of Veterinary Science, The still providing tailored feedback according to University of Sydney, Sydney, Australia, their level of understanding. 2School of Animal & Veterinary Sciences, Charles Sturt University, Wagga Wagga, For access to the module, use the following Australia, 3School of Agriculture and Food link: https://aelp.smartsparrow.com/v/ Sciences, The University of Queensland, ktnsycfu/bu1pjhu3 Gatton, Australia

With the increasing uptake of technology, it OA15.03 Parasites of Zebrafish: Impacts and is important for tertiary educators to move Potential Models (Lemons and Lemonade) towards providing parasitology teaching and learning outcomes across different teaching Dr. Michael Kent1,2, Dr. Thomas Sharpton1, Dr platforms. Parasitologists and veterinarians Katy Murray2, Dr. Justin Sanders1 from several universities throughout Australia 1Oregon State University, Corvallis, United came together to collaborate on an interactive States, 2Zebrafish International Resouce multi-institution module developed to teach Center, Eugene, United States an often neglected aspect of veterinary parasitology to veterinary science students The benefit of aquatic models, led by the in demanding, outcome-driven curriculums. use of zebrafish (Danio rerio), as part of an Our priority was to workshop, build, test, approach to improving human and veterinary deliver and report on a proof-of-principle health is being realized by the scientific flexible teaching module to meet the needs community. As a laboratory animal model, of veterinary parasitology education. The tool it is now second only to mice. The field was chosen was the cloud-based BEST Network/ initially led by investigations in developmental Smart Sparrow platform, which enables the genetics in which experimental end points sharing of purpose-built modules nationally involved primarily embryos or larval fish. or even internationally depending on content, Adult zebrafish are now used extensively as with the capacity to provide response- models throughout the biomedical research triggered feedback at the individual student arena, including infectious disease research. level. Such a platform enables collaboration Naturally occurring pathogens in research on shared resources, with the opportunity facilities, including parasites, are a serious for benchmarking and analytics immediately problem in many facilities as they cause available. The current work showcases our disease and infections in subclinical fish experience with the delivery of a veterinary may cause non-protocol induced variation diagnostics module workshopped in 2018 in experiments. Research laboratories use and developed for deployment to veterinary pathogen-free water as the source for science students in 2019. The interactive aquaria, but they often acquire zebrafish module was designed in collaboration with for studies from pet stores, providing a academics from The University of Sydney, source for many pathogens. The Zebrafish Charles Sturt University and The University International Resource Center has provided a of Queensland, with the support of The diagnostic service to the research community Australian Society of Parasitology. It presents since 2001, and our data from over 100 a theoretical equine parasitology diagnostic laboratories and about 15,000 fish shows scenario that covers themes including that two parasites are quite common in diagnosis, short and long term parasite research facilities. Pseudoloma neurophilia management, communication with clients, (in about 50% of facilities) infects the central and responsible usage of anthelmintic drugs. nervous system is vertically transmitted, it The platform enables students to learn at their may cause emaciation, spinal deformities,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 63 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

reduction in fecundity, and it alters behavior and TBIs (and, when possible, other major endpoints in otherwise clinically normal parasites) of economic and public health fish. Pseudocapillaria tomentosa (reported relevance to SSA. in about 13% of facilities) causes profound intestinal lesions, emaciation, death, and USALTI-Afrique aims to gather data on various lines of evidence indicate that it is the epidemiology and impact of these a promoter of a common intestinal cancer disease systems, documenting the burden in zebrafish. The nematode has benefits as of parasitisms, in several livestock species a model for high throughput anthelminthic across several SSA countries (e.g. Nigeria, drug discover, elucidating interactions of Uganda, etc.). Findings generated will not parasites with bacterial microbiomes, and only be disseminated through the means of cancer research. Last, zebrafish can be specialised scientific reviews and symposia, maintained at human body temperatures, but also shared with recipients from the study and under these conditions we have shown areas (e.g. veterinary and para-veterinary that we can experimentally infect zebrafish professionals, entomologists, local pastoralists with Toxoplasma gondii, and thus providing and livestock keepers, policy makers). another model for parasite drug discovery. The ultimate goal of the group is to guide the design and roll-out of evidence- OA15.04 Training the Next Generation of based strategic control interventions, African Leaders in Parasitology: USALTI- designed according to the specific needs Afrique @ University of Salford of target areas, as well as to help establish contemporary molecular diagnostic capacity Dr. Vincenzo Lorusso1,2, Mr. Babagana in partner laboratories in SSA. Mohammed Adam2, Mr. Adamu Haruna Mamman2, Mrs. Ijeoma Angela Alozor2, Mr At present, the group encompasses several Michiel Wijnveld3, Abraham Goni Dogo4, Dr. postgraduate (PhD and MSc) students as Kevin Bown2, Prof. Richard J Birtles2 well as undergraduate students engaging 1Global Research & Intellectual Property, in summer research projects. Students are Vetoquinol, Paris, France, 2University of trained on the morphological identification Salford Tick Infections (USALTI)-Afrique, of ticks of veterinary and public health School of Environment and Life Sciences, importance, especially from the African University of Salford, Manchester, United continent, as well as on the application of Kingdom, 3Centre for Pathophysiology, a panel of molecular techniques allowing Infectiology and Immunology, Medical the identification of a plethora of tick-borne University of Vienna, Vienna, Austria, infections in biological samples as well as 4Department of Veterinary Parasitology and arthropods. Entomology, Faculty of Veterinary Medicine, University of Jos, Jos, Nigeria This talk provides an overview of the vision and ongoing research activities of USALTI- Ticks and tick-borne infections (TBIs) Afrique, highlighting potential collaboration majorly undermine livestock health, welfare opportunities with groups with shared interest. and productivity in the developing world, including sub-Saharan Africa (SSA). Despite this, there is currently a shortfall of expertise in research and diagnosis of ticks and tick- transmitted pathogens in most of SSA. The University of Salford Tick Infection (USALTI)- Afrique group was established early in 2016 to address this shortage of competences and know-how. USALTI-Afrique provides research, educational and capacity building opportunities to students, scientists and laboratory personnel, focusing on ticks

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 64 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA15.05 Opportunity for Change: OA16 Canine Helminths Implementing AAVMC’s Competency- Based Veterinary Education for Veterinary July 9, 2019, 11:00 - 12:30 Parasitology Plenary Hall, Madison Ballroom (ABCD), Level 4

Antoinette Marsh1, Emma Read1, Andy Moorhead2, Dwight Bowman3, Heather OA16.01 High Risk of False Positive Findings Walden4 by Coproscopy in Danish Dogs 1College of Veterinary Medicine, The Ohio 1 1 State University, Columbus, United States, Dr. Stig Milan Thamsborg , Dr. A. R. Huelle , 1 2 2College of Veterinary Medicine, University of Dr. H. H. Ebbesen , Dr. P. Deplazes , Dr. C.A. 2 1 Georgia, Athens, United States, 3College of Alvarez Rojas , Dr. H. Mejer 1 Veterinary Medicine, Cornell University, Ithaca, Veterinary Parasitology, University of United States, 4College of Veterinary Medicine, Copenhagen, Frederiksberg C, Denmark, 2 University of Florida, Gainesville, United States Institute of Parasitology, University of Zurich, Zurich, Switzerland The American Association of Veterinary Parasitologists (AAVP) Educators Committee The aim of this cross-sectional study was to organizes an educators meeting every other determine the prevalence of endoparasites in year. This meeting provides a unique and health Danish family dogs and to assess the focused forum for those involved in the relative importance of false positive Toxocara- veterinary parasitology curriculum and the egg findings. Through contacts to training training of future veterinarians to meet, discuss clubs in the Copenhagen area, dog owners trends and incorporate various pedagogical volunteered to hand in fresh faecal samples, methods into teaching this subject. In 2017, and 309 dogs ≥4 months were examined we had 39 participants from North American by a modified McMaster (sensitivity of 10 Veterinary Schools or Colleges participate in eggs per g) and a coproantigen(Ag) ELISA a day and a half meeting at the Ohio State (Idexx Laboratories, DK). Toxocara eggs were University College of Veterinary Medicine. speciated as T. canis or T. cati by size and Veterinary Parasitology teaching is not a PCR. Coproscopy detected Toxocara spp. (6.5 static field. Drugs, diagnostic tests, parasite %), hookworm (5.2 %), Cystoisospora spp. (3.6 resistance, and geographic boundaries are ever %), Eucoleus spp. (2.3 %), Taenia spp. (1.0 %), changing, while student demographics are and Strongyloides stercoralis (0.3 %). Non- constantly evolving. To continue to provide canine passengers such as Eimeria spp. and a comprehensive veterinary parasitology Ascaridia galli were also found. The copro-Ag education program, it is critical to review ELISA detected Toxocara (6.9 %), hookworm learning outcomes for veterinary students (10.2 %), Trichuris (0.3 %), and Giardia (18.5 and day-one veterinarians. The American %). Toxocara eggs from 14 dogs were isolated Association of Veterinary Medical Colleges and characterized by PCR (and size) as T. (AAVMC) developed the Competency-Based canis (36%) or T. cati (64%). Agreement by Veterinary Education (CBVE) framework in McMaster and copro-Ag ELISA for positivity March 2018. This framework provides domains was seen in 13 cases while seven dogs were and a series of competencies that describe egg(+)/copro-Ag(-), out of which six dogs a day-one veterinary graduate. Veterinary had eggs identified as T. cati. Eight dogs were Parasitology Educators need to continuously egg(-)/copro-Ag(+) which could perhaps be update their consensus for what constitutes related to pre-patent or low level infections. a “practice-ready veterinarian” and AAVMC’s According to the owners, 45 % of the dogs CBVE framework provides an opportunity for displayed coprophagia. In conclusion, there review. We report on activities at four different was a low prevalence of endoparasites in USA veterinary schools/colleges implementing Danish dogs. The high rate of T. cati eggs the CBVE framework allowing purposeful and other non-canine parasites, associated curricular design related to Veterinary with commonly observed coprophagia, Parasitology. We anticipate additional schools emphasized the importance of false positive and colleges adopting the CBVE framework. results by faecal flotation. False positives may

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 65 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

be reduced by morphological or molecular A. caninum ova shedding. Follow-up fecal identification of eggs, or by only walking examinations were important for verifying the the dog on a leash for 24-48 hours before presence or absence of ova shedding despite sampling. the use of anthelmintic treatment. Limitations of the current study include small sample size, inclusion of only privately owned greyhounds, OA16.02 Combination Anthelmintic and client compliance with fecal collection Treatment for Persistent Ancylostoma and animal care. Caninum Ova Shedding in Greyhounds

Lindie Hess1, Laurie Millward1, Adam Rudinsky1, OA16.03 Comparison of Different Flotation Emily Vincent1, Antoinette Marsh1 Techniques Alone or in Combination 1The Ohio State University College of With ELISA Coproantigen Detection for Veterinary Medicine, Columbus, United States Gastrointestinal Parasites of Dogs

Ancylostoma caninum is a nematode of Dr. Guilherme Verocai1, Dixie Jarriel2, Rita the canine gastrointestinal tract commonly Hanna3, Dr. David Elsemore3 referred to as hookworm. This United States 1Texas A&M University, College Station, United study involved eight privately owned adult States, 2University of Georgia, Athens, United Greyhounds presenting with persistent A. States, 3IDEXX Laboratories Inc., Westbrook, caninum ova shedding despite previous United States deworming treatments. It is unknown how frequently or widespread drug treatment The objective of this study was to evaluate failure and resistance is in dogs because the diagnostic performance of classical there is frequently a lack of subsequent techniques for recovery of parasite eggs routine follow up fecal analysis after normal and oocysts alone or in combination with deworming procedures. Drug resistance a commercial coproantigen ELISAs for in ruminants and equine nematodes is well detecting common gastrointestinal parasites recognized as a significant problem. Drug of dogs. Feces of 132 dogs from shelters and resistant nematodes are not thought to a multidog household were used. Samples be as common in small animals. However, were processed using classical copro- there are reports describing drug resistant parasitological techniques: passive flotation A. caninum and a drug resistant Dirofilaria (PF), single centrifugation sugar flotation immitis strain present in the USA. The dogs (SCSF), and double centrifugation sugar received a combination treatment protocol flotation (DCSF), and the MiniParasep (MP; comprised of topical moxidectin, followed Apacor, UK), a commercial kit whose protocol by pyrantel/febantel/praziquantel within 24 include a centrifugation step. For each test, hrs. At 7 to 10 days post-treatment, a fecal 2g of feces, and Sheather’s sucrose solution examination monitored for parasite ova. (specific gravity = 1.25) were used. Ten Dogs remained on the monthly combination minutes was used for each waiting time and/ treatment protocol until they ceased shedding or centrifugation step(s). Approximately 1g detectable ova. The dogs then received only of sample of 130 of these dogs was stored the monthly topical moxidectin maintenance frozen for coproantigen ELISA detection of treatment. The dogs remained in the hookworms, whipworms, roundworms, and study for 5 to 14 months with periodical Giardia (IDEXX Laboratories Inc.). fecal examinations performed. During the Among the classical methods, DCSF detected study, three dogs reverted to positive fecal more animals infected by Ancylostoma, ova status, with two being associated with Trichuris, and Toxocara (31.1, 32.6, 19.7%, client non-compliance. Re-institution of the respectively), followed by SCSF (29.5, 28.0, combination treatment protocol resulted 18.2%), and PF (28.0, 25.8, 15.2%) and MP in no detectable ova. Use of monthly (27.3, 25.0, 17.4%). Overall, Cystoisospora doses of pyrantel/febantel/praziquantel oocysts, and Taeniidae eggs were detected and moxidectin appears to be an effective in 15.2, and 3.0%, respectively. The ELISA treatment for non-responsive or persistent alone detected more dogs infected with

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 66 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Ancylostoma (38.5%), but less Trichuris 2012, eventually reaching 2.96% by 2018. The (30.0%) and Toxocara (17.7%) than DSCF. highest monthly hookworm prevalence in Among the combinations, DCSF + ELISA 2012 was 2.31%, in May 2012, rising over the detected the most infected animals for 7-year period to a peak summer prevalence Ancylostoma, Trichuris, and Toxocara of 3.22% in August 2018. Each year from 2013 (41.5, 40.0, 21.5%), and found 1, 4, and 3 onward, hookworm prevalence was highest in additional dogs infected for these nematodes, mid-summer and at the lowest in the winter. respectively, when compared to the SCSF Evaluation of gastrointestinal nematode + ELISA combination. Giardia coproantigen prevalence data from over 39 million fecal ELISA detected 38.5% of animals positive, but samples examined over a 7-year period was not assessed using the other techniques. revealed a subtle, yet significant, increasing Among the classical techniques, DSCF prevalence for roundworms (P<0.0001), detected a higher number of dogs infected an increasing prevalence for hookworms with Ancylostoma, Trichuris, and Toxocara; (P<0.0001), and a slightly decreasing and among the combinations, DCSF + prevalence for whipworms (P<0.0001). coproantigen ELISA was superior in the Seasonality was demonstrated for all three detection of these three nematodes. nematodes, possibly for the first time for canine whipworms. Evaluating monthly data 2012-2018: roundworm prevalence was OA16.04 Seasonality and Prevalence of highest December-January and lowest May- Common Canine Gastrointestinal Nematodes June (P<0.0001); whipworm prevalence was in the U.S.A. highest January-February and lowest May- June (P<0.0001); and hookworm prevalence Jason Drake1, Thomas Carey1 was highest July-August and lowest January- 1Elanco Animal Health, Greenfield, United February (P<0.0001). States

Parasite prevalence maps created by the OA16.05 Exploring the Role of ABCB1 Companion Animal Parasite Council (CAPC) Transporters in Toxocara Canis provide monthly data on a county, state and national level, enabling evaluation of Dr. Jeba R J Jesudoss Chelladurai1, Dr. seasonality and changes in prevalence. Matthew T Brewer1 The maps have provided 4.3-7.2 million 1College Of Veterinary Medicine Iowa State canine fecal sample results per year from University, Ames, United States 2012-2018. Changes in prevalence and seasonal fluctuations in canine roundworm, Somatic larval migrans is a feature of Toxocara whipworm and hookworm from 2012 – canis infection in the canine definitive host 2018 were analyzed. Yearly prevalence for and in paratenic hosts. Hypobiotic larval canine roundworm remained between 1.77% pools in the muscles and somatic tissues and 1.94%. Each year, the highest monthly of pregnant bitches are reservoirs for roundworm prevalence of 2.46-2.70% transplacental transmission of infection to occurred during the winter months and lowest multiple litters of puppies. These larval stages prevalence of 1.24-1.47% occurred during the tolerate therapeutic doses of macrocyclic late spring / early summer. Yearly prevalence lactones (MLs) and are difficult to eliminate. of canine whipworm decreased slightly, The mechanism of tolerance to macrocyclic starting at 0.83% in 2012 and gradually lactones in hypobiotic Toxocara canis larvae dropping to 0.67% by 2018. With a seasonal is unknown. However, ML resistance in other pattern similar to roundworm, the highest nematodes of veterinary importance such monthly whipworm prevalence of 0.75-0.95% as Parascaris, Haemonchus, and Dirofilaria occurred each winter and lowest monthly is known to be mediated by ATP binding prevalence of 0.60-0.77% occurred during cassette family B1 (ABCB1) transporters. We the late spring / early summer every year. tested the hypothesis that transport proteins Yearly prevalence for canine hookworm from of the ABCB1 transporters, also known as 2012 to 2018 increased, starting at 2.02% in P-glycoproteins, are involved in the efflux

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 67 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

of the macrocyclic lactones in Toxocara activity was evaluated using Cell Counting canis. We describe the pharmacological Kit-8, and migration and invasion were characterization of the Tca-Pgp-11 transporter determined by wound healing and Transwell and show the localization of Pgp-11 mRNA invasion assays. RHcyst-1 significantly expression using a novel multiple nucleotide inhibited the proliferation, migration, and in situ hybridization technique in adult worms. invasion of all four different tumor cells in We also describe the expression levels of vitro. In addition, a mouse tumor therapy other ABCB1/P-gp genes in various life stages model was established by inoculating the of the helminth and discuss our observations left forelimb of mice with B16-F10 cells, and on the induction of expression of these genes tumor progression was evaluated by assessing in the presence of various MLs. The presented tumor volume and survival. The results data will serve as a primer for further research showed RHcyst-1inhibited tumor growth and on the mechanism of ML tolerance in Toxocara improved survival in vivo. Flow cytometry canis and the use of P-gps as putative was conducted to evaluate myeloid-derived druggable targets for anthelmintic design. suppressor cells (MDSCs), CD4+, and CD8+ T cell levels in PBMCs and spleens. Immunohistochemistry was performed OA17 Molecular Tools I to analyze immune cell infiltration and angiogenesis in the tumors. A decrease and July 9, 2019, 11:00 - 12:30 an increase in MDSCs levels were observed Breakout Room 2, Hall of Ideas E&H, Level 4 in PBMCs and in the spleen, respectively, after RHcyst-1 application. Conclusions: Tick RHcyst-1 has potential antitumor efficacy, OA17.01 A Cysteine Protease Inhibitor and the observed antitumor activities may be RHcyst-1 From Tick Rhipicephalus partly attributable to changes in cathepsin Haemaphysaloides and Its Antitumor expression and MDSCs levels in the PBMCs Potential and spleens. The findings of the present study suggest that RHcyst-1 may have the potential Prof. Jinlin Zhou1, Dr. Nana Wei1, Dr. Yujian to be utilized in cancer treatment. Wang1, Dr. Zhengmao Xu1, Dr. Houshuang Zhang1, Dr. Jie Cao1 1Shanghai Veterinary Research Institute, OA17.02 Evidence of Multiple Point Mutations Chinese Academy Of Agricultural Sciences, in Theileria Annulata Cytochrome B Gene Shanghai, China and Peptidyl-Prolylisomerase Incriminated in Buparvaquone Treatment Failure A potent and tight-binding inhibitor of cysteine proteases had been identified from Dr Bashir Salim1 the tick Rhipicephalus haemaphysaloides, 1University Of Khartoum, Khartoum, Sudan RHcyst-1, which belongs to the cystatin type 1 family and have a predicted molecular weight Drug resistance is one of the emerging and re- of ~11 kDa. An investigation of the RHcyst-1 emerging epidemics affecting both veterinary genes’ expression profile showed that it was and public health sectors. Buparvaquone more richly transcribed in the embryo (egg) provides the most satisfactory means in the stage. The results of proteinase inhibition treatment of bovine tropical theileriosis. assays showed that rRHcyst-1 can effectively However, recently there has been widespread inhibit the six cysteine proteases’ enzyme reports of development of resistance of activities, and especially showed quite a high Theileria annulata to buparvaquone. To inhibitory efficiency against cathepsin L and investigate the situation in Sudan where S, which were considered to be effectively bovine tropical theileriosis is endemic, fifty controlled targets of tumor cells. To blood samples from T. annulata-positive investigate the antitumor effects of RHcyst-1 cattle. were used for DNA extraction, PCR and and to explore the underlying mechanism cytochrome b gene nucleotide sequencing. of these effects, different tumor cells were Analysis of the two buparvaquone binding site treated with RHcyst-1 in vitro. Proliferation regions Q01 (130–148) and Q02 (244–266),

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 68 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

revealed three non- synonymous mutations studies using illumina short reads of three T. at codon 146; alanine (GCT) to threonine orientalis genotypes, Ikeda, Buffeli and a low (ACT) within the Q01 region across all 50 pathogenic genotype (Chitose) have revealed isolates and the other mutation at codon 129; substantial genetic divergence, perhaps at serine (AGC) to glycine (GGC) in 18 isolates the species level. As short read technology which is very close to the Q01 binding site. is unable to effectively resolve the structure However, we documented another mutation of the genomes, we continued to investigate at position 227; valine (GTG) to methionine the isolates using previously generated (ATG) close to the close to the Q02 binding Illumina short reads combined with nanopore site, in three isolates with mutation at codon long reads. In this study, we sequenced the 129. This result was augmented by sequencing three isolates with a R9.4.1 MinION flow cell peptidyl-prolylisomerase gene (targeted by and tested four different hybrid assembly buparvaquone) for 67 samples including the methods, Flye, Canu, Unicycler and Masurca. above 50 samples. Interestingly, we reported Flye and Canu assemblies were further mutation at position 177 in two samples, processed with Nanopolish and five iterations a mutation responsible for buparvaquone of Pilon using Illumina reads. Different treatment failure and one isolate has the combinations of the assemblers were trialed A53P mutation. We showed that this affects and evaluated in order to determine the best Buparvaquone inhibition of Prolyl isomerase pipeline for T. orientalis genome assembly. activity in vitro. In conclusion, we have one Evaluations with Quast and MUMmer revealed interesting confirmation (A53P), several Unicycler to be the best assembler for T. unexplained SP mutations, and two peculiar orientalis Ikeda, and Flye for genotypes, mutations in the STOP codon. This study has Chitose and Buffeli. Alignments to the T. provided evidence of point mutations in the orientalis (Shintoku) reference sequence cytochrome b and peptidyl-prolylisomerase revealed potential structural variation in the genes of T. annulata that is associated with apathogenic Buffeli genotype. The detailed buparvaquone treatment failure in Sudan. methodology and results from this study will be presented and discussed, including the genome annotation and findings of OA17.03 Evaluation of Theileria Orientalis the variation between the pathogenic and Genome Assembly Methods Using Nanopore apathogenic T. orientalis types. Sequencing and Analysis of Variation Between Genomes OA17.04 Novel Subunit Vaccine Approaches Mr. Jerald Yam1, Dr Daniel Bogema2, Dr Cheryl to Protect Against Bovine Babesiosis Jenkins2 1University Of Technology Sydney, Ultimo, Dr Vignesh Rathinasamy1, Dr Carlos Suarez2,3, Australia, 2NSW Department of Primary Dr William Poole1, Dr Heba Alzan2, Dr Marta Industries, Elizabeth Macarthur Agricultural Silva2, Prof Brian Cooke1 Institute, Menangle, Australia 1Biomedicine Discovery Institute, Monash University, Clayton, Australia, 2Department Theileria orientalis (Apicomplexa: of Veterinary Microbiology and Pathology, Piroplasmida) is a non-lymphocyte Washington State University, Pullman, United transforming tick-borne haemoparasite of States of America, 3Animal Disease Research cattle that causes ill-thrift and anaemia. In Unit, United States Department of Agriculture, recent years, clinical outbreaks of T. orientalis Pullman, United States of America caused by the pathogenic genotype of this parasite (Ikeda) have been increasingly Babesia bovis, a major causative agent of observed throughout the Asia Pacific. bovine babesiosis, is a widespread and Currently, there are no available vaccines economically-important protozoan parasite for this disease, although a live vaccine that causes severe, often fatal disease based on a benign genotype (Buffeli) has in cattle. A live vaccine is currently used been proposed to provide cross-protection to partially control bovine babesiosis in against Ikeda. However, our recent genomic endemic areas, however, the variable efficacy

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 69 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

and morbidity associated with live vaccines that can be obtained from foods, and also can warrants alternative, more effective, more easily be formed from phenylalanine catalyzed by deployable and sustainable control measures phenylalanine hydroxylase. Tyrosine is also a such as next-generation sub-unit vaccines. We key starting substrate for producing sclerotin, propose developing novel subunit vaccines that a component of insect cuticle that protect target both, the development of acute clinical insects from pathogen entry. disease and parasite transmission as a novel control approach against bovine babesiosis. Many enzymes are involved in the We are utilising state of the art of genomic, melanization pathway in insects directly proteomic and cell and molecular biology or indirectly, including prophenoloxidase, approaches to define novel potential vaccine dopachrome conversion enzyme, antigens, including parasite antigens involved arylalkylamine N-acetyltransferase, dopa in host cell modification and tick-pathogen decarboxylase, alpha-methyldopa resistant interactions as the basis to identify and develop protein, etc. Mosquitoes have more next-generation vaccines. In cattle infected melanization genes than any other insects, with B. bovis, the parasites cause disease by e.g. Aedes aegypti mosquito has more than altering the structure and function of the red ten prophenoloxidase gene. However, a blood cells (RBCs) that they infect resulting in specific role of each mosquito melanization the unnatural accumulation of RBC’s in various gene is unclear although some biochemical vital organs. We have identified a number of functions have been revealed. uncharacterised parasite-derived exported proteins with predicted functions that are In this report, we used RNAi, CRISPR- pivotal in pathogenesis of severe babesiosis. Cas9 and real-time PCR to investigate Further, we have identified several parasite the roles of some melanization geges in proteins (HAP2, CCp family and 6-Cys family) Aedes aegypti mosquitoes. Several gene that play key roles in sexual reproduction of knock-out mosquitoes have been obtained, the parasite in ticks as additional targets for including a prophenolxidase, dopachrome transmission blocking vaccines. Identification conversion enzyme, and arylalkylamine and biochemical characterisation of selected N-acetyltransferase-1 genes. Obvious vaccine candidates with potential as vaccine phenotypes were found in each mutant line. future candidates for targeting both the asexual Some other genes functions were investigated and sexual stage of B. bovis will presented. using RNAi and real-time PCR, because they are probably lethal genes in the mosquito. The detail findings will be presented in the OA17.05 Understanding Roles of Genes in conference presentation. Mosquito Melanization Response Against Pathogen Infection by Gene Knock-Down and Gene Knock-Out Technologies OA17.06 PCR-Based Distribution of Plasmodium Species in Mosquito Vectors of Mr. Lei Zhang1, Ms Xiaojing Zhu1, Ms Jing Faisalabad District, Punjab, Pakistan Chen1, PhD. Chenghong Liao1, Professor Qian Han1 Mr. Khizer Hayat1, Dr. Muhammad Sohail 1Laboratory of Tropical Veterinary Medicine and Sajid1,2, Prof. Dr. Zafar Iqbal1, Mr. Muhammad Vector Biology, College of Life Sciences and Abdullah Malik1, Mr. Haider Abbas3 Pharmacy, Hainan University, Haikou, China 1University Of Agriculture, Faisalabad, Pakistan, Faisalabad, Pakistan, 2Center for Insect melanization plays roles in defense, Advanced Studies in Agriculture and Food such as wound heeling and native immune Security, University Of Agriculture, Faisalabad, response against pathogen infection, as Pakistan, Faisalabad, Pakistan, 3Department well as roles in cuticle tanning and eggshell of Parasitology, KBCMA, College of Veterinary formation. Melanization is a process leading and Animal Sciences, Narowal Sub-campus of to melanin formation and tyrosine is a key University of Veterinary and Animal Sciences, substrate for melanin synthesis. In insects, Lahore, Pakistan, Narowal, Pakistan tyrosine is a conditional essential amino acid

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 70 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Plasmodium (P.), mosquito-borne unicellular in dogs. Clinical cases of feline leishmaniosis parasite, is responsible for “malaria”. Pakistan caused by L. infantum (FeL) are sporadic but is at risk of malaria because almost 1.6 million increasingly diagnosed. A wide spectrum of cases are reported every year. The present clinical signs is described and associated with study was planned to screen mosquito dissemination of the parasite in skin, mucous vectors for Plasmodium spp. in Faisalabad membranes, lymph nodes, spleen, bone district, Punjab, Pakistan using nested PCR. marrow, eyes, liver, kidney, gastrointestinal For this purpose, convenient sampling of and respiratory tract. However, coinfections adult mosquitoes was done from different (immunosuppressive retroviral infections) places including: animal populated areas, and debilitating diseases (neoplasia, immune- lavatories, water storage tank, livestock farms mediated diseases) are common and may and road side ditches in 70 % ethanol. DNA play a role in the progression of Leishmania extraction was done after stereo microscopic infection to disease and in clinical signs. identification. Species identification of However, evidence-based information is Plasmodium (P. falciparum, P. vivax, P. ovale still limited. Immunopathogenesis in feline and P. malariae) was done through universal infection is scarcely known. Recently, parasite forward and species specific reverse primers specific IFN was demonstrated in infected in the 2nd round of nested PCR. The PCR cats but the role of T cell function remains still products were subjected to agarose gel unclear in felines.γ electrophoresis followed by gel imaging. Prevalence of Culex mosquitoes was higher Suspected cases can be managed according as compared to Anopheles. Plasmodium to LeishVet guidelines formulated by falciparum and P. vivax were found more combining a review of evidence-based prevalent as compared to other species studies, case reports, clinical experience and of Plasmodium. The overall prevalence of critical consensus discussion of LeishVet Plasmodium in mosquito vectors was 46% members. LeishVet guidelines underline (14 out of 30 pools for Plasmodium spp.). the need to confirm a direct role of L. Results were analyzed through chi-square infantum in the clinical manifestations of analyses. Present study may explore the patients and to make a complete clinico- vectorial capacity of mosquitoes which can be pathological evaluation for detecting possible an indicator of Plasmodium distribution in an complications (chronic kidney disease) that area for large scale metagenomics. may affect prognosis. Cytology and histology are the first and best options to confirm FeL. Positive PCR proves the infection and may OA18 Leishmania provide speciation. High parasite loads or antibody levels are suggestive of disease, July 9, 2019, 11:00 - 12:30 but validated serological tools for cats are Breakout Room 3, Hall of Ideas F&I, Level 4 not commercially available. Cats are still empirically treated with the same drugs used in dogs however clinical cure is obtained. OA18.01 LeishVet Symposium: Animal Importantly, on-host repellents are now Leishmaniosis – Lights and Shadows of available to prevent infection of exposed cats Feline Leishmaniosis and they may also reduce transmission from infected cats to sand flies. Maria-Grazia Pennisi1 1University of Messina, Messina, Italy

Cats are infected by Leishmania infantum, Leishmania mexicana, Leishmania venezuelensis, Leishmania amazonensis and Leishmania braziliensis. Leishmania infantum is the most frequently reported species and in endemic areas prevalence of feline infection is not negligible albeit lower than

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 71 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA18.02 LeishVet Symposium: Animal settings. Different techniques may have to be Leishmaniosis – Diagnosis for Veterinarians applied depending on the aim of diagnosing disease or just detecting subclinical infection. Dr. Luís Cardoso1 Additionally, dogs and other animals with 1Department of Veterinary Sciences, School of leishmaniosis might be co-infected with Agrarian and Veterinary Sciences, University further vector-borne pathogens or suffer from of Trás-os-Montes e Alto Douro (UTAD), Vila other concomitant diseases, making diagnosis Real, Portugal more diverse and complicated.

The process of diagnosing leishmaniosis in domestic dogs and other mammalian OA18.03 Toll-Like Receptors 2, 4, and 7, species comprises: (a) confirming disease, i.e. Interferon-Gamma, Interleukin 10 and evaluating leishmanial infections in clinically Programmed Death Ligand 1 Transcripts suspect animals, but also (b) detecting the in Skin From Dogs With Different Clinical presence of Leishmania spp. in subclinically Stages of Leishmaniosis infected hosts. Diagnosis is complex due to the variability of non-specific clinical Dr Laura Ordeix1, Dr. Sara Montserrat-Sangrà1, signs and clinicopathological findings Dr. Pamela Martinez-Orellana1, Dr. Laia compatible with leishmaniosis, and a thorough Solano-Gallego1 diagnostic approach needs to be adapted 1Dep. Medicina I Cirurgia Animals, Universitat for each situation. The disease suspicion Autonoma de Barcelona, Bellaterra, Spain index will be defined by pertinent clinical history, a complete physical examination Canine leishmaniosis is a zoonotic disease and include several routine diagnostic caused by Leishmania infantum that can have tests such as complete blood count (CBC), several dermatological manifestations. The biochemical profile, urinalysis and serum type of immune response elicited against the electrophoresis. The purposes of investigating parasite appears to be at the basis for such infection in clinically healthy animals include clinical variability. However, few studies have epidemiological studies, screening apparently investigated the immunological response in healthy individuals living in areas where the cutaneous lesions in L. infantum naturally leishmaniosis is endemic (e.g. dogs prior infected dogs with different degrees of to vaccination, animals heading towards disease severity. Therefore, the objective of disease progression or breeding dogs), this study was to determine and compare preventing transmission by blood transfusion the transcription of toll like receptors from subclinical carriers, avoiding the (TLRs) 2, 4 and 7, interferon gamma (IFN- ), importation of infected animals to countries interleukin (IL) 10 and programmed cell death where leishmaniosis is not endemic, and protein ligand (PD-L) 1 in paired clinically-γ monitoring response to treatment. Several lesioned and normal-looking skin from 25 techniques have been developed and are diseased dogs (mild disease-stage I (n=11) available, and it is essential to understand and moderate to severe disease-stages II their bases, limitations and appropriate and III (n=14) as well as in normal-looking interpretation, in order to be able to choose skin from healthy dogs (n=10) from a non- the best diagnostic tools for each objective endemic area. Moreover, another objective and subsequently to interpret their results was to correlate the immune response related accordingly. The detection of Leishmania genes transcripts with clinicopathological, infection includes parasitological (cytology, immunological and parasitological findings. histology, immunochemistry and culture), Clinically-lesioned skin from mildly affected molecular (conventional, nested and real- dogs was characterized by a significantly time polymerase chain reaction [PCR]) upregulation of TLR2 (P<0.0001) and IL-10 and serological methods (qualitative and (P=0.021) and downregulation of TLR7 (P= quantitative antibody tests). In addition, 0.004). On the other hand, normal-looking specific cellular immunity tests have also skin of mildly affected dogs was characterized been developed for Leishmania infection, by a significant lower expression of TLR7 but are currently used only in the research (P=0.003), IFN- (P<0.0001) and PDL-1

γ

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 72 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

(P=0.001) and a trend for lower expression symptomatic Ehrlichia (~5.5-fold). NKT cell of TLR2 and IL-10 when compared with more frequency was not modulated. The proportion severely affected dogs. The results of the of NKp46+ NK cells decreased significantly present study provide further insights into the in L+ dogs (~2.3-fold) and in L+E+ dogs TLR and cytokine profile in clinically-lesioned (~5.3-fold), which may signal recruitment of and normal-looking skin of dogs with different this subset to peripheral sites of infection. stages of canine leishmaniosis. In this study, Similarly, the proportion of NKp46+ NKT TLR7 and PDL-1 transcripts were determined, cells decreased (~3.5-fold) in L+E+ dogs. for the first time, in canine skin of dogs Both NK and NKT cells trended to have less with leishmaniosis and they appeared to be intracellular Granzyme B gMFI in L+E+ dogs, associated with disease severity. indicating granule release. In agreement, preliminary studies show both circulating NK and NKT cell frequencies correlate with OA18.04 Altered Circulating NK Cell PBMC cytotoxic activity. Based on these Response During Ehrlichia/Leishmania Co- findings, we hypothesize NK cells from L+E+ infection and Potential Role in Progressive dogs will have increased cytotoxicity and Disease cytokine production compared with EC or L+ dogs, effector functions with important Breanna Scorza1, Kurayi Mahachi1, Erin Cox1, implications for anti-Leishmania immunity. Jennifer Foltz2, Dean Lee2, Jill Saucier3, Phyllis Finally, doxycycline treatment of L+E+ dogs Tyrrell3, Christine Petersen1 with symptomatic Ehrlichia returned NK 1University Of Iowa, Iowa City, United States, cell frequencies to near EC levels, indicating 2Nationwide Children’s Hospital, Columbus, treating comorbid tick infections may United States, 3IDEXX Laboratories, Iowa City, indirectly benefit CanL. United States

Zoonotic Visceral Leishmaniasis (CanL) OA18.05 A Nationwide Survey of Leishmania is driven by transmission of protozoan Infantum Infection in Cats and Associated Leishmania parasites from canine reservoirs to Risk Factors in Italy humans. Identifying factors for development symptomatic CanL is crucial to limiting Dr Roberta Iatta1, Dr Tommaso Furlanello2, transmission and detecting novel human Dr Viviana Domenica Tarallo1, Dr Vito Colella1, immune response targets. We recently Prof Maria Stefania Latrofa1, Prof Emanuele identified a causal association between Brianti3, Prof Paolo Trerotoli1, Prof Nicola tick-borne infection and progression to Decaro1, Dr Eleonora Lorusso1, Dr Bettina CanL. Ehrlichia spp., rickettsia transmitted Schunack4, Prof Guadalupe Mirò5, Dr Filipe by ticks, were among the most common Dantas-Torres6, Prof Domenico Otranto1 tick-borne pathogens in dogs with clinical 1University of Bari, Bari, Italy, 2San Marco CanL (Toepp 2019). How Ehrlichia co- Veterinary Clinic, Veggiano, Padova, Italy, infection alters Leishmania immunity to 3University of Messina, Messina, Italy, 4Bayer lead to CanL progression is unknown. For Animal Health GmbH, Leverkusen, Germany, this study, Natural Killer (NK) cell subsets 5Universidad Complutense de Madrid, Madrid, were compared between endemic controls Spain, 6Instituto Aggeu Magalhães, Fundação (EC) and dogs with CanL +/- Ehrlichia co- Oswaldo Cruz (Fiocruz), Recife, Brazil exposure (L+ and L+E+). We hypothesized Ehrlichia co-infection would be associated Since the first description of leishmaniosis with increased activated NK cells. Flow in a domestic cat (Felis silvestris catus), cytometry was performed on PBMCs for the number of case reports of Leishmania NK (CD3-CD94+ lymphocytes) and NKT infantum infection and clinical cases of (CD3+CD94+ lymphocytes) cell populations. leishmaniosis in cats increased in several Compared to EC, circulating NK cell frequency countries of the Mediterranean basin, with significantly increased in L+ dogs (~2.6- large variability in prevalence data. A major fold), further increased in dogs co-infected limitation in the in comparability of these with asymptomatic Ehrlichia (~3.9-fold) and data has been attributed to the differences

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 73 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

in diagnostic techniques employed and cat infantum and can cause an inflammatory populations sampled. reaction in the gastrointestinal tract, however the role of granulocytic cells (neutrophils, The aim of this survey was to assess the eosinophils, and mast cells) in the intestine prevalence of L. infantum infection in owned of dogs infected is not fully understood. cats across Italy by serological and molecular We performed a quantitative analysis of tests and the identification of potential risk these granulocytic cells in the intestinal wall factors. Blood samples from 2,659 cats of dogs with canine visceral leishmaniasis from northern (n=1,543), central (n=471) (CVL). Twenty dogs were assigned to one and southern (n=645) Italy were tested of three groups: group 1 (G1, n=8), dogs for antibodies against L. infantum, by an with CVL and L. infantum amastigotes in the immunofluorescence antibody test and for intestine; group 2 (G2, n=9), dogs with CVL the parasites’ DNA, by real-time PCR. Samples but without intestinal amastigotes; and group were additionally screened for feline leukemia 3 (G3, n=3), uninfected dogs (control group). virus (FeLV) and feline immunodeficiency Granulocytic cells were counted in the crypt- virus (FIV) proviral DNAs. villus unit, submucosa, and muscle layer of the intestinal mucosa. It was found that in small An overall cumulative L. infantum prevalence and large intestines, the numbers of these of 3.9% was recorded by serology (3.3%) and/ cells increased significantly in dogs of the G2 or by real time-PCR (0.8%), with a higher group followed by the G1 group (p ≤ 0.05), rate (10.5%) in southern Italy. The risk of L. but not in the control group (G3). In G1 there infantum infection in cats was significantly was an inverse correlation between parasite associated to the geographical areas (South burden of the small intestine and granulocytic vs North and Centre; p<0.0001), age class cell counts (r=-0.1; p≤0.01). Exception for (from 19 months to 6 years old vs ≤18 months mast cells in G1 and G2 that were statistically old, p=0.0003), neutering status (not significant only in relation to control group neutered vs neutered, p=0.0028) and FIV (G3) in the large intestine. The granulocytic infection (p=0.0051). cell hyperplasia observed in the intestine of L. infantum-infected dogs suggests that these Though the role of cats in the epidemiology cells may be involved in the cell-mediated of L. infantum is still debated, our findings immune response for parasite elimination. indicate that cats are exposed to and/or infected by this protozoan, mainly in endemic regions of Italy. Hence, as future perspectives, OA19 Nematode Molecular Tools, the standardization of procedures for a Resistance I prompt diagnosis of L. infantum infection and for screening cat populations is a July 9, 2019, 11:00 - 12:30 crucial task for a better understanding of Breakout Room 4, Hall of Ideas G&J, Level 4 the epidemiology of L. infantum infection in cats, and of their potential role in spreading zoonotic leishmaniosis. OA19.01 Experimental Validation of Benzimidazole Resistance Alleles and Mechanisms Using Quantitative Assays of OA18.06 Granulocytic Cells in the Intestinal Free-Living Nematodes Wall of Dogs Naturally Infected by Leishmania Infantum Clayton Dilks1, Qicong Sheng1, Yu-Hsin Huang1, Erik Andersen1 Dr. Wilma Buzetti1, Mr. Diogo Silva1,2, Ms Maria 1Northwestern University, Evanston, United Luana Alves1,2, Dr. Tricia Oliveira2 States 1Unesp, Ilha Solteira, SP, Brazil, 2USP, Pirassununga, Brazil Parasitic nematodes are a major burden of livestock around the world. Anthelmintic Visceral leishmaniasis (VL) is a chronic drugs have been the front line to fight these disease caused by the protozoa Leishmania infections, but resistance arose and spread

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 74 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

quickly worldwide. To fight this resistance, OA19.02 Genetic Characterisation of a thorough understanding of the genetics Benzimidazole Resistance in UK N. Battus and mechanisms of resistance is essential, Populations Using Next-Generation especially for the key benzimidazole (BZ) Amplicon Sequencing and Pyrosequencing class. Three well known BZ resistance variants Technologies (F200Y, E198A, F167Y) are found in a beta- tubulin encoding gene in many nematode Lynsey Melville1, Elizabeth Redman2, Pai Chia species. Additionally, two other resistance Rebecca Chen2, Russell Avramenko2, Alison A. variants have been discovered in nematode Morrison1, Sian Mitchell3, Jan Van Dijk4, Giles parasites at the 198 position, valine and Innocent5, John S. Gilleard2, Dave J. Bartley1 leucine. In addition to variants discovered in 1Moredun Research Institute, Penicuik, parasites, our lab recently identified a number Scotland, 2University of Calgary, Calgary, of other alleles segregating at low frequency Canada, 3Animal and Plant Health Agency, within the free-living nematode species, Carmarthen, Wales, 4University of Liverpool, Caenorhabditis elegans, which are correlated Liverpool, England, 5Biomathmatics and with BZ resistance. To validate whether Statistics Scotland, Edinburgh, Scotland any of these alleles underlie resistance to benzimidazoles, we generated CRISPR/Cas9 Benzimidazole resistance was recently edited strains introducing each of the alleles identified in Nematodirus battus, providing discovered in parasites and C. elegans into an opportunity to create baseline data on the a defined genetic background in C. elegans. prevalence of resistance in this species. As Using high-throughput assays of fecundity with many ovine gastrointestinal nematodes, and growth rate, we quantitatively measured resistance is believed to be conferred by resistance conferred by each of these alleles. single nucleotide polymorphisms (SNP) within In addition, we tested both structurally and the -tubulin isotype 1 gene. Pyrosequencing mechanistically how these alleles affect and deep amplicon sequencing assays beta-tubulin function to confer resistance, usingβ the Illumina Miseq platform were including tissue-specific promoters driving designed to identify the polymorphisms at expression of a susceptible beta-tubulin gene codon F167Y, E198A and F200Y SNPs. The in an otherwise resistant genetic background. aim of the current study was to detect and These tests enabled discoveries of how the quantify BZ-resistant alleles within N. battus drug acts within the animal and confers populations on commercial farms in the UK resistance. Additionally, we investigated and and compare the two technologies. A total experimentally validated how the number of 192 N. battus populations were analysed and diversity of different beta-tubulin genes using each technology. Thirty individual can influence BZ resistance. To the best of parasites were genotyped per population using our knowledge, we have characterized the pyrosequencing and pooled DNA extracts from resistance profile of all currently identified 500-1000 parasites were sequenced by MiSeq. nematode benzimidazole resistance alleles. F200Y was found to be the most prevalent SNP, identified throughout the UK in around 1 in 4 of the populations tested at a low overall frequency of 2.1% ± 0.6% (mean ± S.E.M.). The F167Y SNP was identified for the first time in this species, in four of the populations tested at a low frequency (1.3% ± 0.01%), indicating the early emergence of the mutation. E198A was not identified in any of the isolates tested. Results obtained from pyrosequencing and MiSeq were comparable for F200Y (r2=0.96). The results for F167Y were variable between platforms however, this is likely due to the low allele frequency at this locus (0- 13%). Despite low level variation in results, both technologies were highly comparable and could both be used as a diagnostic tool.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 75 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA19.03 Genotypic Characterisation of susceptible populations suggests that MPTL Monepantel Resistance in UK Isolates of treatment had applied a purifying selection Teladorsagia Circumcincta pressure. Although -tubulin is not the target of MPTL, the process of selecting for Dr. Frank Turnbull1, Prof. Eileen Devaney1, resistance impacts onβ the benzimidazole- Mrs. Alison Morrison2, Dr. Roz Laing1, Dr. Dave resistant genotypes in individuals based Bartley2 on codon 200 mutations. The results 1University of Glasgow, Glasgow, United demonstrate the complexity in characterising Kingdom, 2Moredun Research Institute, genetic resistance in T. circumcincta. Edinburgh, United Kingdom

The preservation of monepantel (MPTL) OA19.04 Diethylcarbamazine Directly efficacy against Teladorsagia circumcincta, Modulates TRP Channels in Filarial the predominant veterinary parasitic Nematodes nematode in UK sheep, is paramount. The first UK case of MPTL-resistance in T. circumcincta Saurabh Verma1, Sudhanva S Kashyap1, was reported in 2018, with unconfirmed Melanie Abongwa1, Alan Robertson1, Richard J evidence of further cases. In this study, we Martin1 had access to a unique resource of three 1Iowa State University, Ames, United States laboratory derived MPTL-resistant strains, three MPTL-susceptible strains from which Canine heartworm disease is caused by they were derived, as well as a collection the filarial nematode Dirofilaria immitis, a of field isolates representing survivors of vector-borne parasitic infection that affects Zolvix-treatment in the previous year. Genetic dogs (definitive host), cats, and in very rare characterisation of individual first and third circumstances, humans. The disease is endemic stage larvae from each T. circumcincta in many parts of the world and infections population were conducted at the Tci-mptl-1 have been reported from every state in loci, -tubulin loci and at ten microsatellite United States. The current recommended loci. A particular focus was on polymorphic regimen for prophylactic treatment with regionsβ in exons 7 and 8 of the Tci-mptl-1 gene macrocyclic lactones (MLs) is not effective at and at mutations F167Y, E198A and F200Y in killing adults. To kill adult worms it requires the -tubulin gene. doxycycline and melarsomine along with MLs and/or surgical interventions, which are The βTci-mptl-1 locus was highly polymorphic high risk and costly. There have also been both within and between populations. reports of heartworm resistance to MLs. The However, comparisons between the MPTL- limited treatment options and development resistant strains and their respective MPTL- of resistance demands identification of new susceptible parental strains showed a drug targets and therapeutic agents. One reduction in the number of genotypes at the approach is to identify the targets of old drugs, Tci-mptl-1 gene. This finding was supported and develop improved compounds for these by low diversity at the Tci-mptl-1 locus in targets. Diethylcarbamazine (DEC) despite its the field isolates post-MPTL treatment. development in 1947 is still used for treatment Pyrosequencing of the -tubulin gene showed of human patients with lymphatic filariasis. a low incidence of the F167Y mutation and no However, the molecular mode of action of evidence of the E198A mutation.β A reduction DEC is undefined with many reports of host- in the number of homozygous benzimidazole- mediated rather than parasite-mediated effects. resistant genotypes and an increase in heterozygosity at position 200 was observed We have studied the direct effects of DEC in the MPTL-resistant strains compared to on our filaria model, Brugia malayi. We their susceptible counterparts. have observed that DEC rapidly produces spastic paralysis, inhibiting adult Brugia The reduction in genetic diversity at the malayi motility. Our follow up results, with Tci-mptl-1 locus in individuals from the MPTL- whole-muscle cell patch-clamp recordings, resistant strains compared to the MPTL- demonstrated that DEC produced a

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 76 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

concentration-dependent inward current SLO-1 channels. We expressed the slo-1 followed by outward current (EC50: 11 M). splice variants heterologously in Xenopus We were able to isolate the inward current oocytes and found that the RCK1 slo-1f by blocking K+ currents: these currentsµ splice variant, found in muscles of males, is were sensitive to TRP antagonists and RNAi more sensitive to emodepside(EC50=5 M) knockdown. than the RCK1 slo-1a splice variant found in muscles of females (EC50>30 M). We alsoμ Our results show that DEC acts by modulating found that emodepside was more potent TRP channels of the parasite. We have also on worms where slo-1a transcriptμ was tested the effects of TRP antagonists/agonists knocked down (EC50(slo-1aKD)=242nM; on adult worm motility. We are presenting EC50(control)=617nM) implying that here, results that support the hypothesis that expression of slo-1a negatively regulates the DEC directly modulates TRP channels of adult emodepside sensitivity in female adult B. filarial worms. malayi. In silico modelling in filarial nematodes revealed that emodepside binds to the Supported by NIH grants: NIH R01 A1047194 cytoplasmic region in 5 binding loops. RCK1 region where alternative splicing occurs in many filarial species contributes four out OA19.05 Emodepside, Macrofilaricide Effects of the five binding loops thereby affecting and Filarial SLO-1 K Channels emodepside potency.

Sudhanva Kashyap1, Saurabh Verma1, Alan Supported by NIH R01AI047194 Robertson1, Richard Martin1 1Iowa State University, Ames, United States OA19.06 Lipidome of Haemonchus Contortus Heartworm infections caused by Dirofilaria immitis are a major concern among dogs and Dr Tao Wang1, Dr Shuai Nie2, Mr Guangxu cats worldwide. The current anthelmintics Ma1, Dr Pasi Korhonen1, Dr Anson Koehler1, Dr regimen for heartworm includes prophylactic Ching-Seng Ang2, Prof. Gavin Reid3, A/Prof. treatment with macrocyclic lactones Nicholas Williamson2, Prof. Robin Gasser1 or multiple doses of doxycycline and 1Department of Veterinary Biosciences, melarsomine to kill adults. Treatment to kill Melbourne Veterinary School, The University adult worms is risky and requires lengthy of Melbourne, Parkville, Australia, 2Bio21 Mass hospitalization and rarely surgery. There Spectrometry and Proteomics Facility, The is a therapeutic gap in administration for University of Melbourne, Parkville, Australia, safe drugs to eliminate adult heartworms. 3Bio21 Molecular Science and Biotechnology Recently, there have been reports of Institute, The University of Melbourne, heartworm resistance to macrocyclic Parkville, Australia lactones. These issues call for a greater need for identification of new drug targets and In spite of progress in the area of lipidomics, develop new and effective interventions that very little is known about lipid biology in do not develop resistance. Emodepside, a parasites. Employing Haemonchus contortus veterinary anthelmintic used for treatment as a model, we characterised the first global of gastro-intestinal nematode parasites, is lipidome for a parasitic nematode using an reported to have macrofilaricidal effects advanced LC-MS/MS-based approach. We against filarian nematodes. We explored the identified > 550 lipid species representing mode of action and sensitivity to emodepside four categories, and explored lipid profiles in using the human pathogen, B. malayi as eggs; third-stage (L3) and exsheathed L3s a filarial model. Worminator motility and (xL3) and fourth-stage (L4) larvae; female and patch-clamp of single muscle cells showed male adults of H. contortus. Lipid composition that emodepside activates voltage-gated and abundance in H. contortus changed potassium channels and that the male is significantly throughput development. The more sensitive than the female. RNAi knock predominant alterations were reflected down demonstrated that emodepside targets in: (i) an increase in glycerophospholipids

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 77 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

(principally glycerophosphocholine and human communities, human behaviours that phosphatidylethanolamine); (ii) a decrease favour parasite transmission, loss of traditional in triradylglycerol synthesis; and (iii) a protective knowledge, and failure to integrate modulation of saturated fatty acids and culturally sensitive messages into well- ether-linked lipids. These changes suggest intended public health risk management. I adaptations in terms of nutrient acquisition, will discuss challenges in assessing, managing, metabolism and development, as H. contortus and communicating risks posed by these transits to the parasitic stage within its host neglected helminth zoonoses. animal. The present lipidomic resources provide a foundation to explore lipid biology in H. contortus and related nematodes, and OA20.02 Factors Associated with to establish the roles of particular in host- Echinococcus Multilocularis Infection in parasite interactions and disease. Coyotes (Canis latrans) in Southern Ontario

Mr Jonathon Kotwa1, Dr. David Pearl2, Dr. Mats OA20 IAFWP Symposium Isaksson3, Dr. Claire Jardine1,4, Dr. Olaf Berke2, Dr. Nicola Mercer5, Dr. Eva Osterman-Lind3, Dr. July 9, 2019, 11:00 AM - 12:30 PM Andrew Peregrine1 Breakout Room 5, Meeting Rooms KLOP, Level 4 1Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Canada, 2Department of Population OA20.01 Emerging Issues With Food- Medicine, Ontario Veterinary College, Borne Helminth Parasites: Trichinella and University of Guelph, Guelph, Canada, Echinococcus 3National Veterinary Institute, Uppsala, Sweden, 4Canadian Wildlife Health Emily Jenkins1, Caroline Frey1,2, Temitope Cooperative, Ontario Veterinary College, Kolapo1, Janna Schurer3, Rajnish Sharma1 University of Guelph, Guelph, Canada, 1University Of Saskatchewan, Saskatoon, 5Wellington-Dufferin-Guelph Public Health, Canada, 2Canadian Food Inspection Agency, Guelph, Canada Saskatoon, Canada, 3Cummings School of Veterinary Medicine and the University of Echinococcus multilocularis was recently Global Health Equity, Kigali, Rwanda reported in wild canids across southern A recent multicriteria-based ranking of food Ontario, a newly recognized endemic area borne parasites ranked Echinococcus spp. for the parasite in Canada. In endemic areas, cestodes and Trichinella spp. nematodes a comprehensive understanding of factors among the top 7 food borne parasites associated with infection in definitive hosts globally from public health and global trade (wild canids) is critical for mitigating risk of perspectives. Even in developed nations transmission to humans and dogs. Evidence with strong capacity for meat inspection suggests that coyotes play an important role and diagnostic testing, these helminth for the maintenance of E. multilocularis in parasites continue to persist, largely due North America, yet little is known about the to wildlife reservoirs. There is evidence transmission dynamics of the parasite in this that E. multilocularis is emerging globally host. A study was therefore carried out to in the circumpolar North (increasing in investigate the association of host-level (sex, prevalence and geographic distribution), body condition), environmental (southern and that unrecognized genetic diversity Ontario region, landcover), temporal (season, within this parasite may be linked to its hunting season, year), and extraneous recent emergence in North America. As factors (submitter type) with E. multilocularis well, there is unrecognized diversity within infection in coyotes in southern Ontario. wildlife-reservoired species of Trichinella, Between November 2015 and March 2017, often of unknown significance for human 416 coyotes were collected from across the health. Other factors driving the emergence region as part of a study that investigated the of these wildlife-reservoired zoonoses include prevalence and distribution of the parasite changing interfaces between wildlife and in wild canids; 24% (95% confidence interval

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 78 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

(CI) 20%–28%) of coyotes were positive for E. programme was based on modern procedures multilocularis. Associations between infection and tools for farm surveillance (by the use of and factors of interest were assessed via a Geographical Information Systems), diagnosis mixed-effects logistic regression model with a in dogs (using the FLOTAC techniques and random intercept for submitter to account for molecular analysis), diagnosis in sheep (by clustering. Coyotes with poor body condition necropsy and ultrasonography), information were at greater odds of E. multilocularis and education (through dissemination of infection than those in good condition (odds brochures, videos, leaflets) for dogs’ owners, ratio (OR) 2.09; 95% CI 1.05-4.14; P=0.036). farmers and school-aged children. A negative association between infection in coyotes and the proportion of natural A ten-year regional initiative addressed the land was observed (OR=0.69; 95% CI 0.54- surveillance (active and passive) and control 0.88; P=0.003). The odds of E. multilocularis of CE from a holistic perspective based infection were greater in government on: 1) geolocation of 306 sheep farms with submitted coyotes compared to those CE infected animals detected by necropsy submitted by hunters (OR 2.94; 95% CI 1.01- and ultrasonography; 2) identification and 8.74; P=0.047). Coyotes from the western treatment of 1166 dogs with praziquantel region of southern Ontario had lower odds in sheep farms (no. = 306) by using ad hoc of infection compared to coyotes from the confinement cages; 3) education for dogs’ central region (OR 0.33; 95% CI 0.15-0.72; owners, farmers and school-aged children. P=0.005). Our findings provide insights into Over ten years, the integrated programme the transmission dynamics of E. multilocularis resulted in a significant reduction of the in coyotes in southern Ontario. infection rates of dogs, sheep and other livestock species in the Campania region of southern Italy. Therefore, this new approach OA20.03 An Integrated Approach to Control represented a valid strategy to control CE in Cystic Echinococcosis in Souther Italy endemic regions.

Laura Rinaldi1, Antonio Bosco1, Maria Paola The procedures and tools developed Maurelli1, Davide Ianniello1, Paola Pepe1, Maria during the programme could be used by Morgoglione1, Leucio Camara Alves2, Giuseppe regional and national health authorities to Cringoli1 target interventions aimed at eliminating 1Department of Veterinary Medicine and CE in animals and humans at fine and large Animal Production, University of Napoli geographical scales. Federico II, CREMOPAR, Napoli, Italy, 2Department of Veterinary Medicine, Federal Rural University of Pernambuco, Recife, Brazil OA20.04 Control of Taenia Solium Transmission by Vaccination of Pigs Cystic echinococcosis (CE) is a severe zoonosis, caused by the larval stage of the Professor Marshall W Lightowlers1 tapeworm Echinococcus granulosus. This 1Faculty of Veterinary and Agricultural helminth infection is of increasing public Sciences, 250 Princes Highway, Werribee, health and socio-economic concern due to Australia the considerable morbidity rates that give rise to high economic losses in the public Taenia solium is a zoonotic cestode parasite health sector and in the livestock industry. which causes human neurocysticercosis. Control programmes against E. granulosus are Pigs transmit the parasite by acting as the considered long-term actions which require intermediate host. A recombinant antigen an integrated approach and high expenditure vaccine, TSOL18, has been developed which of time and financial resources. has achieved very high levels of protection in pigs against experimental challenge The aim of this study was to develop an infections with T. solium. Field trials have integrated approach to control CE in an been completed using the vaccine, including endemic area of southern Italy. The control a cohort trial undertaken in Cameroon and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 79 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

an intervention at population level in pigs In this study we assessed the shedding of in the Banke District of Nepal. In Cameroon, feline lungworms first-stage larvae (L1) and young animals were vaccinated and treated their infectivity to the snail intermediate host once with oxfendazole; controls were given (IH), after anthelmintic administration in cats. oxfendazole alone. At 12 months of age 194 Thirty-six cats diagnosed with lungworm pigs were examined by slicing muscle and infections were enrolled in this study and brain tissue to identify and count parasites. assigned to four groups. In particular, Infection was identified in 19.6% (19/97) of groups A and B were composed of animals non-vaccinated controls while no infection infected by Troglostrongylus brevior and was present among 97 vaccinated animals. treated with eprinomectin (Broadline®) In Nepal, all eligible pigs ≥2 months of age in and moxidectin (Advocate®) formulations, the intervention area were vaccinated with respectively, whereas groups C and D Cysvax, the TSOL18 recombinant vaccine included cats positive for Aelurostrongylus recently registered as Cysvax by Indian abstrusus and treated with eprinomectin Immunologicals Limited, every 3 months and, and moxidectin formulations, respectively. at the same time, given an oral treatment with Prior to and every day after treatment, faecal 30mg/kg oxfendazole. Animals in a region in samples were analysed by the Baermann which no intervention was undertaken served technique and the number of larvae per as controls. Cysticercosis was identified in gram of faeces determined. The efficacy of randomly selected, slaughter weight animals a single administration of the products was at the start and at the end of the intervention finally assessed four weeks after treatment. period. At the start the prevalence of porcine To evaluate the pre- and post-treatment cysticercosis was 23.6% and 34.5% in infectivity of L1 to IH, two snails per cat/day control and intervention areas, respectively. were infected with 100 L1 collected from the Following the intervention, the prevalence faeces of enrolled animals and, then, digested of cysticercosis in pigs from the control area 28 days post-infection. The efficacy of the was 16.7% (no significant change), whereas no eprinomectin and the moxidectin formulations infection was detected after complete slicing at 28 days was 100% for both A. abstrusus of all muscle tissue and brain in animals from and T. brevior, with a mean number of days the intervention area (P=0.004). A three- for cats to become negative of 7.9±1.2 in monthly vaccination and drug treatment group A, 7.8±1.9 in group B, 6.9±1.6 in group intervention in pigs is an effective method C and 8.9±2.0 in group D. Following the for reducing T. solium transmission and, artificial digestion, alive third-stage larvae if implementation where transmission is of T. brevior and A. abstrusus were found in endemic, would be expected to reduce the 160 (87.4%) experimentally infected snails. incidence of human neurocysticercosis. The results of this study demonstrate that a single administration of the two formulations is effective in the treatment of A. abstrusus OA21 Cat Parasitisms and T. brevior infections. Furthermore, during the post-treatment period alive L1 are shed for July 9, 2019, 13:30 - 15:30 up to 8.9±2.0 days and these larvae are still Plenary Hall, Madison Ballroom (ABCD), Level 4 able to reach the infective larval stage in the infected snails. OA21.01 Larval Shedding and Infectivity to Snail Intermediate Host of Feline Lungworms After Anthelmintic Treatment

Maria Alfonsa Cavalera1, Vito Colella1, Rossella Panarese1, Ettore Napoli2, Francesca Arfuso2, Emanuele Brianti2, Domenico Otranto1 1University of Bari, Bari, Italy, 2University of Messina, Messina, Italy

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 80 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA21.02 Vector-Borne Pathogens in bacteria. However, further research is required Domestic Cats From Romania: A First on the implication of various arthropod Epidemiological Survey vector species in their transmission and to understand the pathogenic and zoonotic Dr. Luciana Cătălina Rus1, Dr. Angela Monica significance of these infections. Ionică1, Dr. Ana-Maria Diacu1, Dr. Ana Maria Boncea2, Prof. ECZM Andrei Daniel Mihalca1 1Department of Parasitology and Parasitic OA21.03 Molecular Confirmation of Zoonotic Diseases/University of Agricultural Sciences Strongyloides Stercoralis in Cats and Veterinary Medicine, Cluj-Napoca, Romania, 2FalconVet Veterinary Hospital, Judit Wulcan1, Michelle Dennis1, Jennifer Bucharest, Romania Ketzis1, Thomas Bevelock2, Guilherme Verocai2,3 Different species of bacterial and parasitic 1Department of Biomedical Sciences, Ross vector-borne pathogens can infect felines, University School of Veterinary Medicine, but the role of domestic cats in the Basseterre, Saint Kitts and Nevis, 2Department transmission of various agents to other of Infectious Diseases, College of Veterinary species remains poorly studied. As the data Medicine, University of Georgia, Athens, on the epidemiology of feline parasitic and United States, 3Department of Veterinary bacterial agents is scarce, the purpose of Pathobiology, College of Veterinary Medicine the study was to assess the presence of and Biomedical Sciences, Texas A&M DNA of several vector-borne pathogens in University, College Station, United States cats from Romania. Three hundred fourteen blood samples from cats from various parts Strongyloides felis, Strongyloides planiceps, of the country were included in the study: Strongyloides stercoralis, and Strongyloides owned (n = 267), feral (n = 27), and stray tumefaciens have been reported in cats (n=20). DNA extracted from cat blood worldwide. Differentiation of species has samples was tested by conventional or nested largely been based on geographical location PCR assays for Cytauxzoon/Hepatozoon/ and morphology of larvae retrieved in feces. Babesia/Theileria spp. (18SrRNA), Dirofilaria Location at necropsy has been used to spp. (cox 1), spotted fever group Rickettsiae distinguish S. tumefaciens from the other (gltA), Bartonella spp. (gltA), Ehrlichia spp. species. However, morphometry of these (16S rRNA), Anaplasma phagocytophilum species overlap, with the exception of S. (16S rRNA), Francisella tularensis (17-kDa planiceps, and taxonomic descriptions are lipoprotein gene) and Borrelia burgdorferi poorly detailed for S. felis and S. tumefaciens. s.l. (5S-23S rRNA). DNA of blood protozoans In 2013, 2014 and 2018, under approved was amplified from the blood of 24 cats, IACUC protocols, cats euthanized for health Cytauxzoon sp. being found with the highest reasons and collected as roadside mortalities prevalence (2.9%), followed by Babesia canis on the island of St. Kitts were submitted (1%), Hepatozoon felis (0.6%) and Hepatozoon for pathology and parasite collection. sp. (0.3%). Filarial DNA was found in 2.6% of Strongyloides parthenogenetic females and cats, while Ehrlichia spp. and Bartonella spp. larvae were identified in colonic nodules of were detected in 2.2% and 1.6% of the samples six cats on histopathological examination. The tested, respectively. Moreover, the presence nodules were consistent with those described of Ehrlichia spp. DNA was found to be more in infections by S. tumefaciens. To obtain the frequent in samples collected from stray cats, first genetic sequences of S. tumefaciens, compared to owned or feral cats. None of the DNA from paraffin embedded colonic nodules sampled cats were positive for Rickettsia spp., was extracted and subjected to PCR targeting Borrelia burgdorferi s.l., Francisella tularensis the cytochrome oxidase c subunit 1 (cox1) of or Anaplasma phagocytophilum. Being the the mitochondrial DNA. Phylogenetic analysis first molecular screening of vector-borne revealed that the sequences generated agents in domestic cats from Romania, the belonged to S. stercoralis and clustered in present study showed that cats can harbor a a clade with global isolates assumed to be wide range of blood-associated parasites and zoonotic of humans, dogs, and non-human

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 81 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

primates. These results are the first genetic for the Diagnosis, Treatment and Control confirmation of S. stercoralis in cats and of Feline Endoparasites in the Tropics’. The raise questions about the role of cats in the development of these guidelines required zoonosis of S. stercoralis and the validity of unique and complex considerations to be the species S. tumefaciens. addressed, often inapplicable to developed nations. Much of the tropics encompass middle-to-low income countries in which OA21.04 Addressing Feline Deworming poor standards of environmental hygiene Guidelines in the Tropics and large populations of stray dogs and cats exist. In these regions, endoparasites pose a Filipe Dantas-Torres1,2, Gabriela Pérez Tort1,3, significantly high risk to pets, which in turn Malaika Watanabe1,4, Gad Baneth1,5, Jennifer place their owners at risk of acquiring parasitic Ketzis1,6, Peter Irwin1,7, Andrei Mihalca1,8, Tawin zoonoses. These considerations led to the Inpankaew1,9, Kamani Joshua1,10, Pablo Castro1,11, development of unique recommendations Bui Linh1,12, Arumugam Sangaran1,13, Pablo with respect to deworming and endoparasite Borrás1,14, Rebecca Traub1,15 testing intervals for the control of both 1Tropical Council for Companion Animal global and ‘region-specific’ parasites in the Parasites (TroCCAP), Australia, 2Department tropics. Moreover, the ‘off-’ or ‘extra’-label of Immunology, Aggeu Magalhães Institute use of drugs for the treatment and control of (Fiocruz), Brazil, 3Faculty of Veterinary endoparasites is common practice in many Science, University of Buenos Aires, Argentina, tropical countries and many generic products 4Faculty of Veterinary Medicine, University lack manufacturers’ information on efficacy, Putra Malaysia, Malaysia, 5Koret School of safety, and quality control. Evidence-based Veterinary Medicine, Hebrew University, recommendations surrounding use of such Israel, 6Ross University School of Veterinary drugs and protocols are also addressed in the Medicine, St. Kitts, 7School of Veterinary Guidelines. The formation of these Guidelines Medicine, Murdoch University, Australia, is regarded as the first step towards educating 8University of Agricultural Sciences and and changing veterinarians’ knowledge and Veterinary Medicine of Cluj-Napoca, Romania, perceptions surrounding the veterinary and 9Faculty of Veterinary Medicine, Kasetsart zoonotic significance, diagnosis, treatment and University, Thailand, 10arasitology Division, control of feline endoparasites in the tropics. National Veterinary Research Institute, Nigeria, 11Department of Infectious Diseases, College of Veterinary Medicine, The University OA21.05 Retrospective Fecal Survey of of Georgia, United States, 12Department of Parasites in 2,323 Client-Owned Cats in Parasitology, Vietnam National University of Northcentral Oklahoma From 2007 Through Agriculture, Hanoi, 13Department of Veterinary 2017 Parasitology, Madras Veterinary College, Tamilnadu, 14Administración Nacional de Dr. Yoko Nagamori1, Dr. Mark Payton1, Hadley Laboratoriose Institutos de Salud “Dr. Carlos Apple1, Katherine Herrick1, Dr. Eileen Johnson1 G. Malbrán”, Argentina, 15Faculty of Veterinary 1Oklahoma State University, Stillwater, United and Agricultural Sciences, University of States Melbourne, Australia Our study objective was to determine the prevalence and trend of parasitic infection in The Tropical Council for Companion client-owned cats in northcentral Oklahoma Animal Parasites Ltd. (TroCCAP) is a not- over the past 11 years. All results of centrifugal for-profit organisation whose mission is flotation examination on fecal samples from to independently and freely inform, guide client-owned cats submitted to the Boren and make best-practice recommendations Veterinary Medical Hospital and Oklahoma for the diagnosis, treatment and control of Animal Disease Diagnostic Laboratory of companion animal parasites in the tropics and Oklahoma State University from 2007 through sub-tropics, with the aim of protecting animal 2017 were included. The impact of sex, age, and human health. In line with this mission, seasonality, and years on the prevalence of TroCCAP recently finalised the ‘Guidelines infection were analyzed.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 82 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

A total of 2,323 cases were included for host such as frogs and rodents. To fully this study. Either 33% zinc sulfate solution understand the A. abstrusus lifecycle one (s.g. = 1.18) or Sheather’s sugar solution (s.g. needs to examine infection in the feline final = 1.25) was used to perform a centrifugal host as well as in the intermediate host, the fecal flotation test. Majority of cases (76.1%; mollusk. By studying the intermediate host 1,768/2,323) did not include any parasites, we seek to optimize the infection procedure eggs, oocysts, or cysts. The most common increasing third-stage larvae (L3) production parasite observed was Cystoisospora oocysts by increased efficiency of the snail’s ability (9.1%; 211/2,323), followed by Toxocara cati to maintain the infection. Presented here are eggs (7.6%; 176/2,323), Giardia cysts (4.1%; findings involving the mollusk intermediate 95/2,323), Alaria eggs (3.2%; 74/2,323), host spanning ten years while experimentally Ancylostoma eggs (1.2%; 28/2,323), infecting 34 cats. A. abstrusus positive Dipylidium caninum proglottids/egg packets fecal samples were used to infect aquatic (1.2%; 27/2,323), taeniid proglottids/eggs freshwater snails including Biomphalaria (1.1%; 26/2,323), Demodex mites (0.9%; glabrata, and Cipangopaludina chinensis, as 22/2,323), and Eucoleus aerophilus (0.6%; well as, several species of terrestrial snails 13/2,323). Less commonly, Tritrichomonas including Triodopsis albolabris and Cornu blagburni (0.2%; 4/2,323), Cheyletiella mites aspersum. Third-stage larvae were recovered (0.13%; 3/2,323), Ollulanus tricuspis (0.13%; through pepsin digestion of these snails at 3/2,323), Physalopetra eggs (0.13%; 3/2,323), various time points. This data was used to Toxascaris leonina eggs (0.13%; 3/2,323), assess the infection methods, including the Aelurostrongylus abstrusus larvae (0.09%; snail species, the infection procedure, and the 2/2,323), Platynosomum eggs (0.09%; number of larvae used to infect the snails and 2/2,323), Sarcocystis sporocysts (0.09%; recovered from the snails. In all snail species, 2/2,323), Trichuris felis eggs (0.09%; 2/2,323), there is an overall decrease in the recovery Mesocestoides proglottids/eggs (0.04%; of L3 from the snails the longer the snails are 1/2,323), Otodectes cynotis mites (0.04%; maintained after infection. However, L3 were 1/2,323), Spirometra eggs (0.04%; 1/2,323), still recovered at 357, 453, 464 days post and Toxoplasma-like coccidian oocysts infection from T. albolabris, C. aspersum and (0.04%; 1/2,323) were detected. B. glabrata respectfully. This data furthers the understanding of the A. abstrusus lifecycle in There was no statistical significance between the mollusk, and may provide some insight sexes (p = 0.06); however, a significant trend into what occurs in the mollusk under natural was observed between ages, the younger the field conditions. cats, the higher the prevalence of parasitic infection (p < 0.0001). Statistical analyses also revealed the higher prevalence of infection OA21.07 Factors Associated With the occurred in summer through fall compared to Prevalence of Otodectes Cynotis in winter (p = 0.0005) and overall prevalence of Ambulatory Client Owned Population of Cats infection decreased over years (p < 0.0001). in Pakistan

Dr. Syed Ehtisham-ul-Haque1, Mr. Ali Murtaza1, OA21.06 Aelurostrongylus Abstrusus Dr. Muhammad Arfan Zaman1, Ms. Farah Ijaz1, Infection in the Intermediate Host Mollusk Mr. Muhammad Zeeshan Akram2 Based on the Recovery of Third-Stage Larvae 1University of Veterinary and Animal Sciences, Lahore (Jhang Campus), JHANG, PAKISTAN, Emily Werdal1, Colleen Sorge1, Yuri Lee1, Jhang, Pakistan, 2Animal Nutrition, Omer Janice L Liotta1, Dr. Dwight D. Bowman1 Halisdemir University, Nigde, Turkey, Nigde, 1Cornell University, College of Veterinary Turkey Medicine, Ithaca, United States Aelurostrongylus abstrusus is a feline A study was designed to furnish a base-line lungworm found throughout the world. Its data for prevalence and associated risk factors life cycle requires a mollusk intermediate of O. cyanotis in client-owned cats’ population host and can include a transport or paratenic in Pakistan. The samples were collected

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 83 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

from the cats brought on pet’s clinics and OA22 Molecular Tools II veterinary hospitals for clinical examination in Lahore and Sheikhupura districts of Punjab, July 9, 2019, 13:30 - 15:30 Pakistan. The cats selected for sampling were Breakout Room 2, Hall of Ideas E&H, Level 4 classified on age, sex, breed, housing and intensity of infection. Ear wax of total n=450 cats (n=234 from District Lahore and n=216 OA22.01 Extracellular Vesicles From from district Sheikhupura) were sampled. Ear Hookworms As Vehicles for Vaccine wax were collected by using sterile swabs Candidates and Immuno-Therapeutics soaked in liquid paraffin. Superficial skin scrapings were also taken with the help of Dr Ramon Marc Eichenberger1,2, Dr Javier scalpel blades in those cats that had rashes Sotillo1,3, Prof Alex Loukas1 on skin to check any ectopic mite infestation. 1Australian Institute of Tropical Health and Cats were examined through direct smear of Medicine, James Cook University, Cairns, ear samples using a stereoscopic microscope. Australia, 2Institute of Parasitology, University The intensity (counting) of ear mite parasitism of Zurich, Zurich, Switzerland, 3Carlos III was also made by collecting ear canal Health Institute, Madrid, Spain washings with lukewarm paraffin. Cats were classified on breed, age, sex, housing and Helminth parasites have evolved a vast intensity of infection. Total 159 out of 450 cats array of strategies to manipulate their were found positive for O. cyanotes, showing vertebrate hosts. Extracellular vesicles prevalence of 89/234 (38.03%) in Lahore (EVs) are secreted by all helminth species district and 70/216 (32.40%) in Sheikhupura investigated thus far, and their salient roles in district. Results showed that prevalence was parasite–host interactions are being revealed. highly significantly in cats age of <12months Adult hookworms live in the intestine of (39.67%) than >12months (29.8%). There was the host where they release excretory/ no significant difference present relative to secretory products, representing a major host gender(p>0.05), but a greater number part of the host-parasite interface. While of cases were reported in male cats (36.24%) studies on parasite-host interactions have than females (34.39%). No specific breed traditionally focused on soluble proteins, association to O. cyanotis infestation was we explored the structure, molecular profile found. Prevalence was significantly higher in and functional applications of secreted EVs cats that live outdoor (67.35%), both (outdoor from the rodent parasite Nippostrongylus & indoor) 46.85% with those cats that live brasiliensis, which has been used as a model only indoor 12.29% (p<0.05). All the superficial for hookworm infection. By proteomics skin scrapings were negative to ectopic mite and RNA sequencing, we identified 81 infestation. This is the first study conducted in proteins, including a predominance of SCP- client-owned cats of Lahore and Sheikhupura like proteins, and moreover, hookworm EVs district, Punjab, Pakistan that examined the contain miRNAs that are predicted to bind prevalence of O. cynotis infestations. to murine genes with known roles in the regulation of immunity. Helminths manipulate the host’s immune system towards an immunoregulatory phenotype, which can have beneficial effects for both the parasite and the host. Consequently, there is interest in harnessing the immunoregulatory capabilities of helminths to develop novel therapies for autoimmune, allergic and even metabolic diseases. We have shown that hookworm EVs confer protection against inflammation in models of acute and chronic colitis. Helminth EVs are also of interest as candidates for anthelmintic vaccines, and we have begun to evaluate their protective capacity. Vaccinated

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 84 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

mice with purified N. brasiliensis EVs show One genome and two independent 85.4% reduction in adult worm burden in transcriptomes (one with tissue specific data hookworm challenge infections. We selected and one obtained from in vitro cultured adult and expressed 5 recombinant versions of male worms with and without ivermectin assumed vaccine candidates, presenting exposure) were used as basis for the study. promising but variable efficacies (43.4 – Pgps were characterized using transcriptome- 73.7%). However, vaccination with EVs and guided RT-PCRs and Sanger sequencing, EV proteins did not affect the fecundity and followed by phylogenetic analysis and relative morphology of patent worms. Our studies expression analysis. revealed potential applications of EVs as vehicles in anthelminthic vaccine design, as In total 10 different Pgps were identified. well as in the development of an entirely new Among them, an ascarid specific Pgp lineage, generation of therapeutics to treat chronic Pgp-18, as well as two paralogs of Pgp-11 and non-infectious diseases. Pgp-16. Through re-mapping of transcriptome raw reads on P-glycoprotein cDNAs, relative expression levels were estimated in both OA22.02 Transcriptomic and Genomic transcriptome datasets. Here, no ivermectin Characterization of the P-glycoprotein Gene specific upregulation was found for any of Family in Parascaris sp. the Pgps but Pgp-11.1, Pgp-16.2 and Pgp-9 exhibited significantly higher constitutive Alexander Gerhard1, Dr. Jürgen Krücken1, expression levels compared to low expression Professor Dr. Emanuel Heitlinger2, Professor Pgps, namely Pgp-3, Pgp-12 and Pgp-18. Dr. Jianbin Wang4, Professor Dr. Richard Interestingly, intestinal Pgp expression was Davis4, Professor Dr. Martin Nielsen5, Professor much higher than in any of the other tissues, Dr. Georg von Samson-Himmelstjerna1 e.g. in Pgp-11.1 compared to a whole worm 1Institute for Parasitology and Tropical sample, over 1000 fold. Veterinary Medicine, Freie Universität Berlin, Germany, 2Department of Biology, Molecular The Pgp repertoire of Parascaris sp. comprises Parasitology, Humboldt-Universität zu Berlin, a diverse gene family and several Pgps were Germany, 3Leibniz Institute for Zoo and identified as candidates for future studies Wildlife Research, Research Group Ecology concerning their potential role in anthelmintic and Evolution of Parasite Host Interactions, , efflux. High level Pgp expression in the Germany, 4Department of Biochemistry and intestine suggests a role in intestinal barrier Molecular Genetics, University of Colorado function in nematodes potentially leading to School of Medicine, Denver, United States of reduced uptake of anthelmintics. America, 5Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, United States of America OA22.03 Parasitic Nematode Drug Discovery

Anthelmintic resistance is on the rise in Dr Mostafa Elfawal1, Dr. Sergey Savinov2, Dr. many parasitic nematodes. However, the Raffi Aoian1 mechanisms of anthelmintic resistance are 1University of Massachusetts Medical School, largely unknown. P-glycoproteins (Pgps) have Worcester, United States, 2University of been proposed as putative contributors within Massachusetts, Amherst, United States a multi-genetic context in several nematode species including an important pathogen Parasitic nematodes are amongst the most of foals, the horse roundworm Parascaris prevalent disease-causing agents of human, sp. The complete Pgp gene family of most livestock and companion animals. These nematodes is unkown and to date all studies parasites have a dramatic impact on the have focussed on a few available candidate global economy and together perpetuate the genes. This study aimed to comprehensively poverty cycle via its harmful impact on human characterize the complete Pgp gene family well-being, especially hundreds of millions of in Parascaris sp. at the transcriptomic and children and pregnant women. The cost and genomic level. losses associated with parasitic nematodes

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 85 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

of livestock are estimated to be tens of billions sensitive nematode parasites can lose their of dollars worldwide due to adverse effects on sensitivity to an anthelmintic and recover health and productivity. Parasitic nematodes when they are maintained in the presence of are controlled via regular administration of the anthelmintic. anthelmintic drugs. Unfortunately, parasitic nematodes of livestock are currently resistant to To explore this phenomenon further, we almost all classes of anthelmintic drugs available have been studying effects of maintained today. Following same track, drug resistance concentrations of levamisole on Brugia malayi, in human nematode parasites is underway one of the causes of elephantiasis. We have with multiple reports of reduced activity and observed in Worminator motility studies that warnings for the foreseen resistance disaster. female Brugia initially respond to levamisole Clearly, new drugs against these parasites are with a spastic paralysis; they then progresses highly needed. High-throughput phenotypic to flaccid paralysis; and then recovers to screening is an absolutely essential tool in normal motility after 4 hours exposure to finding new leads for drug development. levamisole. When we have examined effects Parasitic nematodes are not typically amenable of maintained applications of levamisole to to high-throughput screens due to their muscle cells of these worms under patch- large size and the indispensable need for clamp, we find that the inward current laboratory animals for parasites maintenance. responses, due to the opening of muscle Thus, screening against a model organism in nicotinic receptors, decline (desensitize) over place of adult parasites has been commonly a period of 5-15 minutes, which may explain adopted, including the developmental stages the flaccid paralysis that follows the initial of nematode parasites and, most notably, the spastic paralysis. When we examined the free-living nematode Caenorhabditis. elegans. expression of levamisole receptor subunits at To answer whether model organisms are of 4 hours during the recovery, we found that a good representation of adult parasites as unc-38 and acr-8 expression was increased, a screening tool for anthelmintic drugs, we while unc-29 and unc-63 showed little or no screened the drug susceptibility of 1280- FDA- change. In Brugia, there are multiple subtypes approved repurposing drugs to the completion of nicotinic receptors on their body muscle against adult and larval stages of C. elegans and (M-, P-, L, and N-) sensitive to different the hookworm nematode parasite Ancylostoma cholinergic anthelmintics. The increased ceylanicum. Adult whipworms were also tested expression of unc-38 and acr-8 subunits may as part of the protocol. Here we will present allow for an increase of acetylcholine nicotinic the results of our screens, cheminformatics receptors that contribute to the recovery of prioritization, in vivo validation, and our motility; but not the recovery of sensitivity proposed pipeline for anthelmintic drug to levamisole that requires the presence of development. We will also present progress receptors composed of UNC-38, ACR-8, UNC- towards automation of the pipeline and 29 and UNC-63 subunits. delineation of a high-throughput platform. Thus, we can see that some worms have dynamic mechanisms that allow them to OA22.04 Dynamic Mechanisms Allow adjust their sensitivity to an anthelmintic. Worms to Adjust Their Sensitivity to an It will be interesting to determine how Anthelmintic: Levamisole and Brugia Malayi widespread these mechanisms are in our parasites. Dr Richard John Martin1, Mr Mengisteab Wolday1, Dr Sudhanva S Kashyap1, Dr Saurabh Supported by NIH R01AI047194 Verma1, Dr Alan P Robertson1 1Iowa State University, Ames, United States

Anthelmintics can become ineffective if they develop resistance, which is usually explained as being acquired over generations due to genetic selection. However, individual, initially

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 86 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA22.05 Anthelmintic-like Activity of LL-AQ showed an EC99 of 0.462 ± 0.017 mg Leucaena Leucocephala Aqueous Extract mL -1, with a dose-dependent behavior (P < Against Gyrodactylus Spp., and Its Effect 0.05). Histopathological analysis revealed no over the Host Health damage treatment-associated, discarding a negative impact that could jeopardize the Dr Elke von Son-de Fernex1, MVZ Juan E. life of treated animals. The aqueous extract Compean-Martínez1, IAZ Martha Salazar-Ulloa1, of L. leucocephala showed to be a promising M en C Biol Larisa Chavéz Soriano2, M en C alternative to control gyrodactylus infestation Biol. Germán Muñoz-Córdova1 in Tilapia fish. 1Centro de Enseñanza Investigación y Extensión en Ganadería Tropical, Facultad de Medicina Veterinaria y Zootecnia, Universidad OA22.06 In Silico Analysis of Potential Nacional Autónoma de México, Tlapacoyan, Vaccine Candidates for Tritrichomonas Mexico, 2Departamento de Patología Foetus, the Causative Agent of Bovine Veterinaria de la Facultad de Medicina Trichomoniasis Veterinaria y Zootecnia de la Universidad Nacional Autónoma de México, Distrito Ms. Eleanor Senior1, Dr Andrew Jackson1 federal, México 1Institute Of Infection and Global Health- University of Liverpool, Liverpool, United The monogean parasite Gyrodactylus spp has Kingdom been considered one of the most aggressive and pathogenic parasites in aquaculture; Tritrichomonas foetus is an anaerobic and the use of formaline baths has been flagellated protist and the causative agent of the cornerstone for gyrodactylid control. the venereal disease bovine trichomoniasis. However, formalin has been been reported a This disease causes spontaneous abortions carcinogenic and mutagenic agent for both and, in some cases, infertility in cows and is host and consumers. The use of bioactive responsible for decreased calving rates and plant has been recently proposed as an milk production; infected animals are usually alternative against gyrodactylus infestation. culled. Bovine trichomoniasis is therefore Leucaena leucocephala is a tropical plant responsible for significant economic losses to reported with anthelmintic activity against farmers in several countries where the disease both internal and external parasites of cattle. is endemic, including Australia, Brazil and the Therefore, the objective of this study was USA. Currently there is no vaccine available to assess the anthelmintic-like activity of that can prevent reinfection. Leucaena leucocephala aqueous extract (LL-AQ) against Gyrodactylus spp., in tilapia In order to identify potential vaccine fish, and the effect over the host health. candidates for this parasite a reverse Sixty-three naturally infected tilapia fish vaccinology approach was implemented. The (± 40 parasites per fish) were divided in 7 Tritrichomonas foetus genome was sequenced experimental groups (n=3) and placed in on the PacBio platform (147Mb, N50 = 84,706), experimental fish restraint-chutes for a 60 assembled using SMRTportal and then min bath treatment. Five concentrations of annotated using multiple automated processes LL-AQ were employed: 2, 1, 0.5, 0.25, 0.125 mg including BRAKER, SNAP and BLAST2GO, / mL -1. Distilled water and formalin (1:4000) integrated with transcriptomic data from both were used as negative and positive control, trophozoite and pseudocyst cell types, and respectively. Three replicates were run for improved through manual curation. Cell surface each concentration and control. Parasites specific genes were identified using in silico were counted pre and post-treatment using prediction of signal peptides, transmembrane a stereomicroscope. Effective concentration domains and GPI anchors. (EC99) was calculated through a Probit Analysis using a maximum likelihood In our T. foetus genome 84,706 genes have estimation method (CI 95%). Finally, the been identified, 1,607 of which contain a sixty-three animals were humanely slaughter signal peptide and a transmembrane helix to assess for histopathological changes. The suggesting they are cell surface expressed

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 87 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

and will be further examined as potential voltage operated Ca2+ channels (VOCCs). epitopes. Once a reduced set of genes has been Involvement of VOCCs in MCLZ’s mechanism produced, they will be recombinantly expressed is supported by data showing that MCLZ and tested for their immunogenic potential. is (i) phenocopied by a VOCC agonist, (ii) Ca2+ dependent and (iii) inactive in worms We have produced the first fully-annotated T. with a deletion in a key VOCC subunit. foetus genome as the first step in a reverse MCLZ also causes collapse of mitochondrial vaccinology approach to this important membrane potential and promotes cleavage livestock disease. Preliminary analysis of of proapoptotic caspases, supporting a predicted cell surface proteins has resolved model whereby VOCC activation increases diverse transmembrane proteins as potential intracellular Ca2+, exceeding mitochondrial vaccine candidates. buffering capacity to drive permeability- pore transition and programmed cell death. Collectively, these data show that the actions OA22.07 Mechanism of the Benzodiazepine of the benzodiazepine MCLZ on parasitic Meclonazepam (Ro 11–3128) on Parasitic flatworms are distinct from PZQ and provide Flatworms a foundation for developing analogs with improved parasite selectivity. Dr. John Chan1 1Medical College of Wisconsin, Milwaukee, United States OA22.08 Must Improved Animal Welfare Lead to More Parasites? Parasitic flatworm infections (e.g. tapeworms and fluke worms) are treated by Ms. Emelie Pettersson2, Dr Marie Sjölund1, a limited number of drugs. In most cases, Dr Eva Osterman Lind1, Professor Johan control is reliant upon praziquantel (PZQ) Höglund2, Professor Per Wallgren1 chemotherapy. However, PZQ is ineffective 1National Veterinary Institute, Uppsala, against larval/immature worms (cestode Sweden, 2Swedish University of Agricultural cysts and sexually immature lung/liver stage Sciences, Uppsala, Sweden schistosomes), and there have been several reports on the emergence of PZQ-resistant The national parasite status of pigs in Sweden cestodes and trematodes. Given the need has not been investigated since the 1980’s. for alternative therapies to treat parasitic Since then, new animal welfare laws have flatworm infections, we have revisited the been implemented, stating that pigs are to anthelmintic benzodiazepine meclonazepam be kept loose at all occasions and sows are (MCLZ). MCLZ was discovered in the 1970’s often housed on deep litter straw beds. Slats but not brought to market due to dose- are also only allowed to constitute 30% of the limiting sedative side effects. We were pen floor. It has been hypothesized that these intrigued by observations that MCLZ differs examples of more natural housing conditions from PZQ in several key aspects and have may increase the parasite load. resumed studies on trematodes to determine its molecular mechanism of action. We show The aim of our study was to investigate the that MCLZ is effective against juvenile, PZQ- status of gastrointestinal parasites in 25 herds refractory Schistosoma mansoni infections with more than 100 sows. In 2017-2018, a total of in vivo. MCLZ also kills parasitic flatworms 962 faecal samples were collected from piglets directly in vitro, while PZQ’s effects are (n=205), growers (n=207), fatteners (n=177) rapidly reversible following washout and and sows (n=373) and analysed for nematode its mechanism is likely immune-dependent. eggs and coccidian oocysts using a modified The targets of PZQ and MCLZ appear to be McMaster technique based on 3 g of faeces. distinct, which augurs well for the utility of a benzodiazepine based-therapy should reports Parasites were found in all but one herd. of PZQ-refractory parasites become more In total, Ascaris suum was detected in 6%, frequent. Binding assays show that MCLZ, but Oesophagostomum spp. in 19%, Trichuris suis not PZQ, displaces radioligand from L-type in <1% and coccidian oocysts in 13% of the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 88 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

samples. Compared to the 1980’s, Ascaris Leishmania-infected, dogs. These alterations suum decreased from 27% to 8% in fatteners, in follicular organization will be associated but increased from 7% to 11% in sows. Also, with changes in population size and Oesophagostomum spp. decreased from 6% distribution of splenic CD4+CXCR5+TfH cells to 3% in fatteners but increased from 27% to and CD19+PDL1+ Breg cells. 43% in sows. Eimeria spp. were found mainly in sows, 17% compared to 2% in the 1980s. In Dogs were selected from an outbred cohort piglets Cystoisospora suis was detected in previously shown to have L. infantum infection 16% compared to 20% in previous studies. and determined to be asymptomatic or symptomatic through physical examination. Overall our results showed a comparably These dogs were humanely euthanized for low parasite load in fatteners, indicating tissue collection. Light microscopy was used that the changed housing conditions are not to evaluate splenic structure from H&E stained associated with increased parasite exposure, slides. Staining on slides was secondarily despite increased contact with manure analyzed with digital imaging software to and a limited use of chemical disinfectants measure follicle area, total white pulp area, and antiparasitic drugs. A possibility is and total red pulp area. that age-segregated rearing, that has been implemented in most herds since the 1980’s, From these initial results we conclude that decreases especially the spread of Ascaris when measured by digital imaging software, suum from older to younger pigs. adult symptomatic dogs exhibited a greater total follicle area than adult asymptomatic dogs. Adult asymptomatic dogs also had OA23 Leishmania a higher ratio of primary (immunologically dormant) to secondary (immunologically July 9, 2019, 13:30 - 15:30 active) follicles. Juvenile asymptomatic dogs Breakout Room 3, Hall of Ideas F&I, Level 4 exhibited a higher manual follicle count/ mm2 of splenic area than adult symptomatic or adult asymptomatic dogs. These results OA23.01 Splenic Architecture Alterations suggest greater immunologic activity in adult and Cell Phenotypes between Symptomatic symptomatic dogs, and age-related differences and Asymptomatic US Dogs with Visceral in follicle size and activation within the cohort. Leishmaniasis

1, Breanna Scorza1, Kurayi Mahachi1, Erin Cox OA23.02 Safety and Efficacy of Canileish Angela Toepp1, Diogo Valadares1, Katherine Vaccine in Exposed Dogs Gibson-Corley1, Christine Petersen1 1University of Iowa, Iowa City, United States Dr. Manuela Gizzarelli1, Dr. Valentina Foglia Manzillo1, Dr. Fabrizio Vitale2, Dr. Salvo Sotera3, Dogs are the reservoir species for Dr. Serena Montagnaro1, Prof. Gaetano Oliva1 visceral leishmaniasis, and canine visceral 1Department Of Veterinary Medicine and leishmaniasis closely models the course Animal Production, University of Naples of disease in human patients. Secondary “Federico II”, Naples, Italy, 2Istituto lymphoid organs, particularly spleen, Zooprofilattico Sperimentale della Sicilia – undergo alterations in immune function and CreNaL, Palermo, Italy, 3DVM, Lampedusa microscopic structure as VL progresses from asymptomatic to clinically symptomatic Aims stages. How and why these changes occur Vaccination against CanL reduces the parasite are not well understood. We hypothesized load and the risk of progressive infection in that dogs with symptomatic canine visceral dogs. The study aimed to investigate the risk leishmanaisis will have fewer splenic follicles, of developing clinical disease in exposed dogs poorly organized follicles and germinal (Paltrinieri et al.,2010) when vaccinated with centers, and lower splenic white pulp LiESP/QA-21 (CaniLeish™) and to assess if it is area when compared to asymptomatic, safe in these animals.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 89 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Contents 1Federal Rural University of Pernambuco, Recife, The study was performed during two Brazil, 2Ceva Santé Animale, Paulínia, Brazil consecutive transmission seasons in Lampedusa, a small Italian island highly In the New World, Leishmania infantum is endemic for canine leishmaniasis (Foglia the agent of visceral leishmaniasis and CanL, Manzillo et al.,2018). Sixty-nine owned dogs of which are spread by the bite of infected different sex, breed and age were submitted sand flies. The dog is considered the main to clinical examination, blood sample and reservoir in urban areas of Brazil and the vaccination. The presence of anti-Leishmania treatment of CanL still represents a challenge. antibodies was assessed by a rapid The objective of this study was to evaluate immunochromatographic test (Speed Leish the clinical and laboratory aspects of dogs K™, Virbac BVT) before the prime vaccination. naturally infected with L. infantum submitted Sera were analysed by IFAT too. Dogs were to therapy with marbofloxacin associated monitored every six months and after one with allopurinol. To evaluate the efficacy of year they received the booster vaccination. marbofloxacin 12 domiciled dogs of both Vaccinated dogs were divided in: group genders, varied race and age between 1-7 A - 43 negative to the pre-vaccination IFAT years were used. The animals were divided test; group B - 26 positive to IFAT at low titer into two treatment groups: Group 1 (G1) were (<1:160). No statistical difference was found treated with oral marbofloxacin (Marbopet® when the age was compared (P=0,2696 - Laboratory CEVA, Brazil) at 2 mg/Kg/day for Kendall’s Tau). Only 4 animals (2.7%), two 28 days in combination with oral allopurinol for each group, showed high antibodies level at 10 mg/kg every 12 hours and Group 2 (G2) and clinical alterations. Kaplan-Meyer survival treated with marbofloxacin and allopurinol curves demonstrated no statistical difference in the same dosage, but the allopurinol between the groups (P=0,5516). At the end was give to dogs from G2 after 28 days of follow-up 9 dogs (20,93%) of group A of administration of marbofloxacin. After remained negative to IFAT while the others the treatment dogs were assessed during showed low antibody titers (≤ 1:160). In group 90 days by performing monthly physical B, 1 dog converted to negative serology, exams, and evaluating red blood count, white the others maintained initial values. Sixty- blood count, alkaline phosphatase, alanine five dogs had no clinical abnormalities. In aminotransferase, urea, creatinine, serum both groups few transient adverse events to protein, albumin and globulin parameters. vaccine were recorded. Each clinical parameter was classified according to its severity on a numerical Conclusion scale of 0 to 3. In group G1, the reduction Our data seem to demonstrate that: i) the in scores was 76.4%, while the G2 group CaniLeish™ vaccine is safe also in dogs presented a reduction of 54.8%. The averages classified as “exposed” (Paltrinieri et al.,2010) of albumin were higher at 90 days in the or “subclinically infected” (Solano-Gallego et G1 group. No dogs from both of groups al., 2009); ii) in these dogs CaniLeish™ avoids presented increase of urea and creatinine. the progression of the Leishmania infection After the following up of 90 days it can be during two consecutive transmission seasons. concluded the marbofloxacin and allopurinol treatment in combination at the same time provided a greater remission of clinical OA23.03 Evaluation of Clinical, Laboratory signs, improvement in hematological and Profiles of Dogs Naturally Infected With biochemical profiles and increase of albumin L. Infantum Submitted to Therapy With value. Marbofloxacin Associated With Allopurinol

Dr. Leucio Alves1, Dr. Janilene Nascimento1, Dr. Claudio Rossi2, Dr. Leonardo Brandao2, Dr. Talita Lins1, Dr. Maria Vanuza de Meireles1, Dr. Wagner Andrade1, Dr. Roseane Feitosa1, Dr. Winny Silva1, Dr. Edna Michelly Santos1

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 90 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA23.04 Evaluation of DPP-Leishmania and test had sensitivity of 96.3% and specificity SNAP-LeisEvaluation of DPP-Leishmania of 100%. Agreement between both antibody and SNAP-Leishmania Rapid Tests for tests and parasitological detection methods Diagnosis of Leishmania Infantum Canine was 96.8%. The DPP® test had sensitivity of Infectionshmania Rapid Tests for Diagnosis 95.8% and specificity of 100%. The SNAP® of Leishmania Infantum Canine Infections and DPP® tests demonstrated high and similar levels of sensitivity and specificity. The SNAP Celeste Souza1, Professor Valmir Silva2, Dr. test can be used for screening canine samples Bruno Alberigi3, Professor Norma Labarthe4 for Leishmania antibodies. 1Laboratório de Imunomodulação e Protozoologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, OA23.05 Canileish Vaccine and Repellents Brazil, 2Faculdade de Medicina de Campos, Are Effective Measures to Prevent Canine Fundação Benedito Pereira Nunes, Campos, Leishmaniasis in Field Condition: A 6-Year- Brazil, 3Programa de Pós Graduação em Field Retrospective Study Medicina Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, Brazil, Dr. Antonio Inglese1, Dr. Manuela Gizzarelli2, Dr. 4Programa de Bioética, Ética Aplicada e Saúde Valentina Foglia Manzillo2, Dr. Gaetano Oliva2, Coletiva, Fundação Oswaldo Cruz, Rio de Dr. Christelle Speiser Fontaine3 Janeiro, Brazil 1Private veterinary clinician, Grottaglie, Italy, 2Università degli Studi di Napoli Federico Visceral leishmaniasis is a zoonotic disease II - Dipartimento di Medicina Veterinaria e of global importance that affects up to 1 Produzioni Animali, Napoli, Italy, 3Virbac, million humans every year, causing at least Carros, France 20,000 deaths worldwide. Besides affecting humans, it also affects wildlife and domestic Medical preventative measures against species. Since dogs play a key role in urban leishmaniosis includes the regular use of Leishmania spp. transmission, the Brazilian phlebotomine repellents and/or of vaccines government maintains a Program to Monitor (Maroli et al., 2010). The study aimed to assess and Control Visceral Leishmaniasis (PVCLV), the efficacy of these measures in long term which promotes awareness campaigns aiming field conditions. to enhance control of the infection. The PVCLV recommends the DPP® canine visceral Data were collected between February, 1st leishmaniasis test (Bio-Manguinhos, Brazil) for 2012 and April, 30th 2018 in a veterinary clinic screening and an ELISA to confirm infection. located in Grottaglie, Italy. Dogs of ≥ 6 months The DPP® test is produced and distributed by of age at the time of the leishmaniosis diagnosis the Health Ministry to the Municipal Health or of the first vaccine injection were allocated Centers responsible for the local PVCLV; the into 4 groups: no preventative measure (NVNR), product is not available to clinics, forcing repellents only (NVR), vaccine only (VNR), most veterinarian practitioners to use other vaccine and repellents (VR). Leishmaniosis rapid tests for screening and diagnosis of this diagnosis was confirmed if dogs were disease in their clinical routine. This study was symptomatic with ELISA = 1, or, when ELISA conducted to compare the performance of > 1 (Solano-Gallego et al., 2009). Cases with the DPP® and the SNAP® Canine Leishmania uncomplete data were not analyzed. Antibody Test (IDEXX Laboratories, Inc., USA) tests using sera from dogs with confirmed The analysis was made on 240 dogs: 31/240 infections of L. infantum as well as sera from (13%) in group NVNR, 63/240 (26%) in dogs residing in non-endemic areas or in group NVR, 63/240 (26%) in group VNR areas with a low prevalence of Leishmania and 83/240 (35%) in group VR. The groups infection. The results obtained showed were not different in terms of sex, breed, that SNAP® and DPP® tests were virtually life style repartition or repellents used when equivalent for detection of canine antibodies appropriate. The dogs of the VR groups were against L. infantum. There was 97.0% statistically younger than those of the NVR agreement between the two tests. The SNAP® and the NVNR groups at the beginning of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 91 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

the follow-up (respectively 1.4 ± 0.8, 2.9 ± dogs were followed for a minimum of six 1.5 and 3.5 ± 2.8 years of age). Diagnosis of months. Dogs belonged to 20 different breeds leishmaniosis was confirmed in 20/31 (34%) and the mean age was 3,9 years. The mean dogs for the NVNR group, 10/63 (15.9%) dogs time between vaccination and diagnosis of in the NVR group, 0/63 (0%) in the VNR and CanL was 10 months (range: 4 to 16 months) 0/83 (0%) in the VR group. and the majority of cases (41/46 dogs, 89%) had been treated with topical insecticides- CaniLeish™ vaccine (Oliva et al., 2014; Martin repellents. At time of diagnosis, 4 dogs were et al., 2014; Moreno et al., 2014), in addition or considered to have the disease in stage I not to repellents, prevented the development (Leishvet-classification), 33 in stage II, 8 in of an active Leishmania infection in all stage III and 1 in stage IV. Twenty-three dogs (146/146) dogs. Unvaccinated dogs treated (50%) had low-medium IFAT titers, and the with repellents were 9.6 time less likely rest had medium-to-high titers. In most cases to develop a leishmaniosis infection than (33/46,72%), dogs received the standard untreated dogs. Vaccination and/or the use of treatment (N-methyl-glucamine-antimoniate repellents decreased the risk of leishmaniosis or miltefosine and allopurinol). After six in at-risk dogs. months, 43/46 dogs (94%) were still alive and 78% were free of clinical signs. Anti- Leishmania antibody titers (IFAT) dropped OA23.06 Leishmaniosis in Vaccinated Dogs: considerably and 33/35 (94%) were negative Clinical Aspects or had low-medium titers.

Rubén Foj1, Glòria Pol1, Jerónimo Carnés3, Pilar Altogether, these data seem to indicate that Brazis1, Lluis Ferrer2 the disease in vaccinated dogs presents a 1LETIPharma, Animal Health, B.U., Barcelona, mild clinical course. Spain, 2Animal Medicine and Surgery Department, Faculty of Veterinary Medicine, UAB, Bellaterra, Spain, 3LETIPharma, R & D OA24 Nematode Molecular Tools, Allergy & Immunology, Madrid, Spain Resistance II

The approval by the EMA of two vaccines July 9, 2019, 13:30 - 15:30 against canine leishmaniosis (CanL) has been Breakout Room 4, Hall of Ideas G&J, Level 4 a major step in the control of this zoonosis in Europe. In recent years, hundreds of thousands of dogs have been vaccinated. However, OA24.01 Identification and Biochemical the impact of vaccination on the disease Characterization of Three Conserved epidemiology and on the health of individual Haemonchus contortus Cathepsin B-like dogs has yet to be evaluated. According to Proteases published data both licensed vaccines have an efficacy of protection close to 70%, meaning Dr. Mariam Bakshi1,2, Dr. Dante Zarlenga1 that some vaccinated dogs develop the 1USDA-ARS, Beltsville, United States, 2Oak disease. The characteristics of the disease in Ridge Institution for Science and Education, vaccinated dogs is of major importance. Oak Ridge, United States

The objective of this study was to characterize The trichostrongyle, Haemonchus contortus, the clinical aspects of leishmaniosis in dogs is a blood sucking abomasal nematode that vaccinated against the disease with LetiFend® causes significant economic losses due to (Protein Q-vaccine). treatment costs and reductions in animal health and productivity. Incorrect and Forty-six vaccinated dogs were diagnosed excessive chemical deworming have resulted with CanL based on clinical signs, in anthelmintic resistance to all major classes clinicopathologic abnormalities (complete of drugs. Consequently, effective vaccine blood count, serum biochemistry, urinalysis) targets continue to be sought as cheap and and positive serology (ELISA and IFAT). All alternative measures for control. Parasite-

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 92 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

derived proteases and protease inhibitors have costs and poses a serious threat to farm been implicated in controlling host responses animal health. To prevent the establishment to infection and therefore make good vaccine of resistant strains of this parasite, up- candidates. In-silico analyses of protease and to-date molecular techniques need to be protease inhibitor sequences from H. contortus proposed which would allow for quick, cheap showed high degrees of conservation among and accurate identification of individuals clade V nematodes. ‘Conserved Domain infected with resistant worms. The effort has Analysis’ (NCBI) was performed followed by been made in the previous decade, with the homology modeling using SWISS-MODEL to development of the pyrosequencing method identify a class of cysteine proteases known to detect resistance-predicting alleles. as Cathepsin B-like (CBP) which are essential Here we propose a novel droplet digital parasite virulence factors and therefore PCR (ddPCR) assay for rapid and precise attractive vaccine targets. Further, analyses identification of H. contortus strains as being showed that all identified proteins possessed resistant or susceptible to benzimidazole canonical CBP active sites and classic drugs based on the presence or absence secretory signals. To this end, three novel CBP of the most common resistance-conferring genes from H. contortus, designated Hc-CBP-1, mutation F200Y (TAC) in the β tubulin isotype Hc-CBP-2 and Hc-CBP-3, were selected, cloned 1 gene. The newly developed ddPCR assay and expressed. The full-length Hc-CBP-1, Hc- was first optimized and validated utilizing CBP-2 and Hc-CBP-3 genes are 1044, 1041 and DNA templates from single-worm samples, 1050 base pairs in length and encode proteins which were previously sequenced using the that are 348, 347 and 350 amino acids long, next generation PacBio RSII Sequencing respectively; sequence homology spanned (NGS) platform. Subsequent NGS results 57-67% among the proteins. When expressed for faecal larval cultures were then used as in a prokaryotic system, the protein masses a reference to compare the obtained values ranged from ~35-37 kDa via gel analysis and for fractional abundances of the resistance- Western blots. We investigated the functional determining mutant allele between ddPCR and degradative roles of these recombinant and NGS techniques in each sample. Both cathepsins on various protein substrates over a methods managed to produce highly similar broad pH range in vitro. Our analyses showed results and ddPCR proved to be a reliable tool that these enzymes are likely multifunctional which, when utilized at full capacity, can be and may be involved in nutrition, development used to create a powerful mutation detection and pathogenicity of H. contortus. These and quantification assay. results suggest that H-CBPs are promising candidates for developing vaccines against Haemonchosis. OA24.03 Transcriptomic and Genomic Approaches to Ivermectin Resistance in Haemonchus Contortus OA24.02 Exploring Benzimidazole Resistance in Haemonchus Contortus by Next Generation Dr. Roz Laing1, Ms. Kirsty Maitland1, Dr. James Sequencing and Droplet Digital PCR Cotton2, Dr. Stephen Doyle2, Ms. Nancy Holroyd2, Dr. Collette Britton1, Dr. Dave Bartley3, Paulius Baltrusis1, Dr. Peter Halvarsson1, Ms. Alison Morrison3, Prof. Eileen Devaney1 Professor Johan Höglund1 1University Of Glasgow, Garscube Estate, 1Swedish University of Agricultural Sciences, Bearsden Road, United Kingdom, 2Wellcome Uppsala, Sweden Sanger Institute, Hinxton, United Kingdom, 3Moredun Research Institute, Edinburgh, Anthelmintic resistance in gastrointestinal United Kingdom nematode (GIN) parasites of grazing ruminants is on the rise in countries across the The BUG consortium is a UK based world. Haemonchus contortus is one of most project aimed at using the Haemonchus frequently encountered drug-resistant GINs in contortus genomic resources to improve the small ruminants. This blood-sucking abomasal understanding of anthelmintic resistance, nematode contributes to massive treatment with an emphasis on ivermectin (IVM). Using

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 93 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

a genetic cross between a drug sensitive threat to the sustainable control of species (MHco3) and a drug resistant isolate that parasitize humans. Unfortunately, (MHco18), a locus on chromosome V was the genetic basis of resistance to most shown to be under selection by pooled whole anthelmintics remains poorly resolved. As genome re-sequencing of F3 L3 pre- and a key aim of the BUG Consortium, we have post-IVM exposure. Transcriptomic analysis used a forward genetics approach together was also carried out using male and female with whole genome sequencing to unravel worms from both parental isolates and the the genetic mediators of multidrug resistance F2 generation of the cross, with and without in the gastrointestinal parasite, Haemonchus IVM exposure. As expected there were many contortus. We performed a genetic cross constitutive differences in gene expression in between susceptible MHco3(ISE) and triple the parental samples (MHco3 versus MHco18). resistant (benzimidazole, levamisole & However, genes that were also differentially ivermectin) MHco18(UGA) parasites, after expressed in pairwise comparisons of the which we sampled the F3 population both F2 generation of the cross with and without pre- and post-treatment of the F2 population IVM exposure represent a high confidence in vivo for each of the three drug classes. set of genes associated with IVM resistance. For each of the drug classes, we identify Many of the differentially expressed genes discrete, non-overlapping regions of the had orthologues in C. elegans and these were genome linked to resistance. We identified significantly enriched for neuromuscular a number of previously characterized genes GO terms, including regulation of neuronal linked to benzimidazole and levamisole differentiation (males) and contractile resistance; however, for ivermectin, a fiber (females). In an attempt to prioritise more complex signature of selection was candidate genes in the genomic locus under apparent, which included a major driver on IVM selection, the transcriptomic data was chromosome V as well as a number of minor used to identify genes that also showed effect loci contributing to resistance. Finally, differential expression in IVM-resistant adults. we demonstrate how underdosing primes These included a number of kinases and the genetic landscape, leading to increased several genes involved in regulating neuronal/ frequency of some alleles before higher dosing behavioural plasticity. The orthologues of selects unique combinations of mutations to these genes are now being studied in C. produce more resistant individuals. These data elegans to determine whether they do indeed provide an important genome-wide overview play a role in IVM resistance. of the genetic response to anthelmintic treatment, and an insight into the evolution and spread of resistance. OA24.04 Unraveling the Genetic Mediators of Multi-Drug Resistance in Haemonchus Contortus Using Forward Genetics OA24.05 How to Detect Genomic Signatures of Selection Associated With Anthelmintic Dr. Stephen Doyle1, Dr. Roz Laing2, Dr. David Drug Resistance in Field Populations of Bartley3, Dr. Alison Morrison3, Dr. Kirsty Parasitic Nematodes Maitland2, D.r Umer Chaudhry4, Dr. Nancy Holroyd1, Dr. Matthew Berriman1, Dr. John Dr. Janneke Wit1, Dr. Matthew Workentine1, Gilleard5, Dr. Neil Sargison4, Dr. James Cotton1, Dr. Umer Chaudhry2, Dr. Sam Yeaman3, Dr. Dr Eileen Devaney2 Sean Rogers3, Dr. James Wasmuth1, Prof. John 1Wellcome Sanger Institute, Hinxton, United Stuart Gilleard1 Kingdom, 2University of Glasgow, Glasgow, 1Faculty of Veterinary Medicine, University United Kingdom, 3Moredun Research Institute, Of Calgary, Calgary, Canada, 2University of Penicuik, United Kingdom, 4University of Edinburgh, Edinburgh, Edinburgh, 3Faculty Edinburgh, Edinburgh, United Kingdom, of Biological Sciences, University of Calgary, 5University of Calgary, Calgary, United Kingdom Calgary, Canada Drug resistance in parasitic helminths is The ability to detect genomic signatures of widespread in a number of domesticated selection in field populations of parasitic animal systems, and represents a genuine nematodes would allow the genetic basis of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 94 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

resistance to both current and new drugs to OA24.06 Investigating the Molecular be more rapidly determined and resistance Epidemiology of Benzimidazole Resistance to be detected at an earlier stage. The in Ovine Gastrointestinal Nematodes Using recent completion of a chromosomal level Deep Amplicon Sequencing reference genome assembly for Haemonchus contortus now makes this potentially feasible. Dr. Camila De Queiroz1, Dr. Russell Avramenko1, However, the nature of genomic signatures Dr. Elisabeth Redman1, Dr. Fabienne Uehlinger2, of selection associated with drug resistance, Dr. Michel Levy1, Dr. John Gilleard1 and the best way to detect them, is poorly 1University Of Calgary, Calgary, Canada, understood. Consequently, we have used H. 2University of Saskatchewan, Saskatoon, contortus as a model to investigate these Canada questions using field populations predicted to have strong genomic signatures of selection Anthelmintic resistance is a threat to associated with benzimidazole resistance. sustainable parasite control worldwide. An We have undertaken two different genome- understanding of how resistance mutations wide scanning approaches to compare H. emerge and spread is foundational contortus populations from government information for strategies aimed at prolonging sheep flocks in Pakistan, subject to frequent the useful life of current drugs. To date, benzimidazole treatment and closed to animal sequence polymorphisms in codons 167, movement, with pastoral flocks that are rarely 198 and 200 of the isotype-1 -tubulin gene treated. We genotyped ~100,000 polymorphic have been associated with benzimidazole markers across the 300Mb H. contortus resistance of several gastrointestinalβ genome from 15 individual adult worms per nematode species. New sequencing population using double-digest Restriction- technologies now allow us to study the site Associated DNA (ddRAD) sequencing. molecular epidemiology of these mutations We also undertook whole genome sequencing in the field in unprecedented scale. Deep of 15 individual worms from the same amplicon sequencing, using next-generation populations. A strong signature of selection sequencing platforms, has many advantages, centred on the isotype-1 -tubulin locus was including the ability to detect low frequency visible in the drug resistant populations using resistance mutations and haplotypes with nucleotide diversity, linkage disequilibrium a high scalability due to the ability to and Fst analysis. Weaker signatures of multiplex hundreds of samples and loci in a selection were apparent at a number of single sequencing run. We have undertaken other genetic loci. We have used the data to a large-scale screen for benzimidazole compare the relative merits ddRADseq versus resistance-associated mutations of the major whole genome sequencing and of “Poolseq” gastrointestinal nematode species and applied versus individual worm sequencing. This phylogenetic analysis to study the origins work provides a strong proof-of-concept that and spread of these mutations in the region. genome-wide scanning can unequivocally Populations of L1 larvae were harvested detect anthelmintic resistance associated from sheep fecal samples, collected before signatures of selection in field populations of and after treatment with benzimidazoles, parasitic nematodes and provides important from more than 40 farms across western information as to the best approaches. Canada. We sequenced the relevant region of the isotype-1 -tubulin locus and applied PEAR (paired-end read merger) to the reads obtained from eachβ sample to form contigs. We then used the Mothur bioinformatic tool to assign sequences to nematode species and a pipeline with BWA, VarDict and SnpSift to identify non-synonymous mutations occurring at the three codons. Finally, we calculated the frequency of each of the resistance associated single nucleotide polymorphisms. The F200Y allele was present at high frequency

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 95 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

in many H. contortus, T. circumcincta and T. one scaffold was under selection in both our colubriformis populations. We will present an experimental system and a natural population. analysis of the association of this mutation However, it was not the only scaffold under with the resistant phenotype, evidence of selection and other scaffolds showed a selection and haplotype network analysis stronger association with TCBZ-resistance to investigate the origin and spread of this in this population. This may mean there are mutation in Western Canada. multiple loci associated with TCBZ-resistance or that, given the inherent genetic variation present in field populations compared to our OA24.07 Does Triclabendazole Resistance controlled clonal genetic cross, the results are in Fasciola Hepatica Have the Same Genetic more complex to interpret. Basis in Different Areas of the UK? These results provide a basis to define the Dr. Nicola Beesley1, Miss Rebecca Hoyle1, mechanism of TCBZ-resistance. Professor Diana Williams1, Professor Steve Paterson2, Professor Jane Hodgkinson1 1Veterinary Parasitology, University of OA24.08 A Comprehensive Study of Liverpool, Liverpool, United Kingdom, 2Centre N-Glycosylation of Haemonchus Contortus for Genomic Research, University of Liverpool, Liverpool, United Kingdom Dr. Chunqun Wang1, Dr. Wenjie Gao2, Dr. Xin Liu2, Dr. Min Hu1 Fasciola hepatica causes parasitic disease 1Huazhong Agricultural University, Wuhan, in livestock. Emergence of resistance to China, 2Huazhong University of Science and triclabendazole (TCBZ), the treatment of Technology, Wuhan, China choice, is a substantial threat to control; yet the mechanism of TCBZ-resistance is unknown. N-glycosylation is one of the most prominent Using an experimental genetic cross of clonal post-translational modifications of proteins susceptible and resistant F. hepatica, we that plays pivotal roles in varieties of have determined that a single locus, ~3.2Mb biological functions and processes. The in size, can confer TCBZ-resistance and is studies in N-glycosylation of helminths dominantly inherited. However, we do not have attracted attentions not just due know if resistance has the same genetic basis to the characteristics of composition of in geographically distinct natural populations. N-glycosylation, but also for their potential roles in vaccine and new drug development. We sourced parasites from two populations: Haemonchus contortus is an economically (i) adult F. hepatica collected from six sheep important blood-sucking nematode of experimentally infected with a TCBZ-resistant ruminants with worldwide distribution. population from Wales, UK, (FhLivR4pop); Previous evidences have indicated that three of which were TCBZ-treated and three several bioactive and immunogenic proteins untreated (ii) F. hepatica eggs collected in H. contortus are highly glycosylated. from Day 0 and Day 21 samples following However, little is known about the nature of composite faecal egg count reduction test N-glycosylation of H. contortus, especially (cFECRT) on five farms from Wales and in this area of functional N-glycoproteomics. England, UK. Following Illumina sequencing, Here, we comprehensively mapped we mapped reads and identified regions N-glycosylation of adult H. contortus using with significant differences in SNP allele high-resolution mass spectrometry-based frequencies between TCBZ-treated and glycomics and proteomics, as well as lectin untreated, to highlight genomic regions histochemistry to locate these glycoproteins associated with TCBZ-resistance. on the H. contortus anatomical sites. Results showed that high mannose glycans were The ~3.2Mb region identified in our genetic dominated in the N-glycan profiling that crossing experiment was shown to be were primarily expressed on intestine, associated with TCBZ-resistance in adult gonad of H. contortus and lectin localization parasites from FhLivR4pop. This indicates that analysis confirmed the abundance of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 96 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

N-glycosylated proteins. We further enriched numbers of cysts and oocysts into the the glycoproteins using hydrophilic interaction environment. These animals are thought to chromatography with mass spectrometry and play a significant role in the contamination of identified 559 N-glycosylated sites on 337 foods, especially fresh produce, potentially proteins expressed in H. contortus. Among leading to illnesses and outbreaks. these identified proteins, a large proportion of Contamination of fruits and vegetables may proteins are involved in metabolic processes. result through livestock accessing crop lands According to the sequence alignment or surface waters, agricultural runoff, or the analyses, 14 potential vaccine candidates use of improperly treated manure as fertilizer. with known immune modulatory properties Products of animal origin, such as milk and were detected including aminopeptidases, meat, may also become contaminated due to zinc metallopeptidases, cysteine proteinases, poor udder hygiene or fecal contamination galectin and pepsinogen. In addition, we also at the slaughterhouse, respectively. Further revealed the glycosylation characteristics of work is required to clarify the role of many key molecules involved in the growth livestock in the contamination of foods development and invasion processes of this with zoonotic isolates of G. duodenalis and parasite. This study provides a comprehensive Cryptosporidium spp. A better understanding insight into the N-glycosylation composition of the potential sources of contamination of H. contortus, suggesting the importance will be required in order to develop and of further explorations in examining implement effective control strategies. This glycosylation in the development of novel presentation will discuss the potential role interventions against haemonchosis. of livestock in the contamination of foods with G. duodenalis and Cryptosporidium spp., and its public health significance. The OA25 IAFWP Symposium discussion will be illustrated using the results of studies performed in our laboratory on the July 9, 2019, 13:30 - 15:30 prevalence and molecular characterization Breakout Room 5, Meeting Rooms KLOP, Level 4 of these parasites in cattle and swine, as well as in different food commodities, including packaged leafy greens, shellfish, and fresh OA25.01 The Role of Livestock in the meats purchased at retail. Foodborne Transmission of Giardia Duodenalis and Cryptosporidium Spp. To Humans OA25.02 Host Adaptation in Cryptosporidium Parvum: Genetic Basis and Dr. Brent Dixon1 Implications to Epidemiology 1Health Canada, Ottawa, Canada Yaoyu Feng1, Yuanfei Wang1,2, Dawn Roellig2, Giardia duodenalis and Cryptosporidium spp. Professor Lihua Xiao2 are common protozoan parasites in humans 1Key Laboratory of Zoonosis, Ministry of and domestic and wild animals worldwide, Agriculture, College of Veterinary Medicine, and are important causes of enteric illness. South China Agricultural University, Transmission occurs by means of the fecal- Guangzhou, China, 2Division of Foodborne, oral route, whereby G. duodenalis cysts and Waterborne, and Environmental Diseases, Cryptosporidium spp. oocysts are ingested Centers for Disease Control and Prevention, directly, through contact with feces, or Georgia, United States indirectly by means of a vehicle such as water or food. While person-to-person and Cryptosporidium parvum is the waterborne transmission account for most Cryptosporidium species with the broadest human infections, zoonotic transmission has host range, responsible for most zoonotic also generated considerable interest. Both Cryptosporidium infections in humans. Host G. duodenalis and Cryptosporidium spp. adaptation, however, is known to occur within are highly prevalent in livestock, particularly the species based on sequence analysis young animals, which can shed very large of the hypervariable 60 kDa glycoprotein

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 97 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

gene, with IIa, IIc, and IId subtype families only Cryptosporidium parvum is a serious preferentially infecting calves, humans pathogen. Prevalence studies indicate a and lambs, respectively. To improve our high occurrence of C. parvum infections in understanding of the genetic basis of host pre-weaned calves. Molecular methods for adaptation in C. parvum, we sequenced the species identification are now widely used in genomes of 257 specimens of the IIa, IIc, and epidemiological studies of Cryptosporidium IId subtype families from various sources infections. Additionally, amplification and using the Illumina 250 × 250 bp paired-end sequencing of the 60-kDa glycoprotein (gp60) technique. Comparative genomics analyses gene provides the basis for characterization have identified significant differences in of C. parvum at a subtype level. However, to nucleotide sequences among subtype families better understand transmission dynamics and of C. parvum. There are ~13,000 single to answer many unresolved epidemiological nucleotide polymorphisms (SNPs) between questions, it is necessary to improve detection IIa and IId subtype families and ~20,000 SNPs of mixed subtype infections. Highly sensitive between IIa and IIc subtype families across tools with good resolution power are needed the 9.1 Mb genome. Most highly polymorphic to better understand mixed infections. genes among the three subtype families Therefore, a method to study intra-host are subtelomeric ones encoding secretory subtype diversity using next generation proteins, especially the invasion-associated sequencing (NGS) of the gp60 gene on and immunodominant mucin proteins and the Illumina MiSeq platform was developed members of the Cryptosporidium-specific and used to expose C. parvum subtype SKSR gene family. The three subtype composition in fecal samples from calves. families also differ in the copy numbers of The results demonstrate the suitability of this subtelomeric genes encoding the MEDLE assay to improve our understanding of C. family of secretory proteins and insulinase- parvum subtype diversity. They also denote like proteases, with IIc having significantly that subtyping studies able to elucidate intra- fewer copies than IIa and IId. Geographically host subtype diversity are required to better segregated subpopulations have been further understand host specificity, source of infection, identified in the IIa and IId subtype families at and transmission dynamics of C. parvum. the whole genome level. Thus, host adaptation in subtype families of C. parvum is associated with significant genomic differences, and the OA25.04 Cryptosporidium Genotypes and occurrence of host adaptation in the major Subtypes in Diarrheal Dairy Calves in France zoonotic Cryptosporidium species is likely to reduce the occurrence of zoonotic infections Dr. Mohamed Mammeri1, Aurélie Chevillot3, in humans and cross-species transmission in Ilham Chenafi1, Myriam Thomas3, Christine farm animals. Julien2, Isabelle Vallée3, Bruno Polack1, Jérôme Follet4, Prof. Karim Adjou1 1UMR BIPAR, Ecole Nationale Vétérinaire d’Alfort, OA25.03 Improving Cryptosporidium ANSES, INRA, Université Paris-Est, Maisons- Parvum Subtype Identification Using Next Alfort, F-94700, France., Maisons-alfort, France, Generation Sequencing 2Phileo Lesaffre Animal Care, 137 rue Gabriel Péri, 59 700 Marcq-en-Barœul, France, 3UMR BIPAR, Dr. Monica Santin1, Dr. Jenny Maloney1 Anses, Ecole Nationale Vétérinaire d’Alfort, INRA, 1EMFSL/ARS/USDA, Beltsville, United States Université Paris-Est, Animal Health Laboratory, Maisons-Alfort, F-94700, France, 4Université de Cryptosporidial infection is one of the most Lille, CNRS, ISEN, UMR 8520- IEMN, Lille France common causes of diarrhea in humans and 59000, France livestock worldwide. The epidemiology of cryptosporidiosis has received significant Little is known about the genetic of attention because of the public health Cryptosporidium in calves in eastern regions and economic importance of this disease of France. The aim of this study was to detect in humans and animals. Out of the four and isolate Cryptosporidium spp. in faecal species predominantly identified in cattle samples from naturally infected pre-weaned

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 98 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

calves (≤ 45 days-old) in France. A total of 35 Marcq-en-Barœul. France, 4UMR BIPAR, diarrhoeic pre-weaned calves faecal samples Anses, Ecole Nationale Vétérinaire d’Alfort, were collected from 26 dairy cattle farms INRA, Université Paris-Est, Animal Health with or without diarrhoea in 6 provinces. The Laboratory, Maisons-Alfort, F-94700, France, screening was established microscopically by 5Université de Lille, CNRS, ISEN, UMR 8520- the detection of Cryptosporidium oocysts using IEMN, Lille France 59000, France an immunofluorescence (IF) staining method. Little is known about the presence of IF-positive samples were then analysed to Cryptosporidium spp. and Giardia duodenalis determine species by PCR-RFLP and sequencing in Algerian sheep, nor their potential role targeting the 18S rRNA gene. C. parvum positive as zoonotic reservoirs. This study aimed samples were subtyped through the analysis of to investigate the occurrence of these two the partial 60 kDa glycoprotein (gp60) gene. protists in lambs. A total of 83 fecal samples Data were then integrated into a phylogenetic were collected from lambs (< 40 days old) tree analysis. IF revealed the presence of on different 14 farms. Samples were screened Cryptosporidium oocysts in 31 out of 35 (88%) for Cryptosporidium spp. and G. duodenalis samples. Combining results of 18S rRNA gene presence with Immunofluorescence technique analysis, C. parvum was detected in 30 samples. (IF). Nested PCR of the small subunit Subtyping analysis in 27/30 samples (90%) of ribosomal RNA (rRNA) gene, followed by the C. parvum isolates revealed two zoonotic restriction fragment length polymorphism subtype families, IIa (24/27) and IId (3/27). Four (PCR-RFLP) and sequence analyses were subtypes were recognized within the subtype used to identify Cryptosporidium species. family IIa including IIaA15G2R1 (21/27) which Then, C. parvum was further subtyped by is the hypertransmissible subtype the most sequencing the highly polymorphic 60kDa frequently reported worldwide, IIaA17G3R1 glycoprotein (gp60) gene. For G. duodenalis, (1/27), IIaA17G1R1 (1/27) and IIaA19G1R1 (1/27). nested PCR of the glutamate dehydrogenase Two subtypes were recognized within the IId (gdh) and triose phosphate isomerase (tpi) subtype family including IIdA22G1 (2/27) and genes were applied and then PCR-RFLP was IIdA27G1 (1/27). These findings illustrate the high used to determine G. duodenalis assemblages. prevalence of Cryptosporidium in calves in dairy Cryptosporidium oocysts and Giardia cysts herds and increase the diversity of the molecular were detected in 36/83 (43%) and 23/83 characteristics of C. parvum isolates with the (28%) of fecal samples, respectively. Of first description of IIaA17G3R1, IIaA19G1R1 and the 21/36 (58%) Cryptosporidium samples IId subtypes in France. The presence of zoonotic that were IF-positive, 16/21 (76%) were subtypes of C. parvum species suggests that identified as C. parvum, and 5/21 (24%) C. pre-weaned calves are likely to be an important ubiquitum. From 15 C. parvum isolates, 2 reservoir for zoonotic C. parvum. subtypes were identified within the subtype family IIa including IIaA21G2R1 (3/15) and IIaA13G2R1 (1/15), while IIdA16G1 (11/15) was OA25.05 Cryptosporidium and Giardia the only subtype within IId subtype family. Species and Genotypes in Sheep in Algeria Of the 16/23 (69%) G. duodenalis IF-positive samples, the most frequent assemblage was Dr. Lynda Sahraoui2, Myriam Thomas1, Aurélie the ruminant-specific assemblage E (10/16), Chevillot1, Dr. Mohamed Mammeri1, Dr. Bruno followed by assemblage D (4/16), and A Polack1, Dr. Isabelle Vallée1, Dr. Jérôme Follet5, + E mixed assemblages (2/16). This study Prof. Hacina Ain-Baaziz2, Professor Karim reports for the first time the identification and Adjou1 genotyping of both Cryptosporidium spp. and 1UMR BIPAR, Ecole Nationale Vétérinaire Giardia duodenalis from lambs in Algeria. This d’Alfort, ANSES, INRA, Université Paris-Est, is also the first description of assemblage D Maisons-Alfort, F-94700, France, Maisons- in sheep. The presence of zoonotic C. parvum Alfort, France, 2Laboratory of Animal Health subtypes (IIa, IId) and C. ubiquitum, as well as and Production, Ecole Nationale Supérieure G. duodenalis, indicates that sheep could play Vétérinaire (ENSV) d’Alger, Issad-Abbes an important role as a potential reservoir for Oued-Smar, Algiers, Algeria, 3Phileo Lesaffre zoonotic protists. Animal Care, 137 rue Gabriel Péri, 59 700

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 99 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA25.06 Seroprevalence and Associated OA25.07 Serological Detection of Risk Factors of Toxoplasma Gondii Infection Toxoplasma Gondii in Marine Mammals From in Domestic Animals in the Oliver Reginald Canada Tambo District, South Africa Adrian Hernandez Ortiz1, Brent Wagner1, Dr. Whatmore Tagwireyi1, Prof. Eric Etter2, Rajnish Sharma1, Dr Batol Al-Adhami2, Dr Prof. Luis Neves3 Pierre-Yves Daoust3, Enooyaq Sudlovenick3, Dr 1University Of Pretoria, Onderstepoort, South Sonja Ostertag4, Dr Lisa Loseto4, Dr Nicholas Africa, 2CIRAD, UMR Animal, Santé, Territoires, Pilfold5, David McGeachy5, Nicholas Lunn6, Dr Risque et Ecosystèmes (ASTRE), France, Emily Jenkins1 3Centro de Biotecnologia Universidade 1Department of Veterinary Microbiology, Western Eduardo, Mozambique College of Veterinary Medicine, University Of Saskatchewan, Saskatoon, Canada, 2Centre Toxoplasma gondii is a protozoan parasite that for Food-borne and Animal Parasitology, infects birds and mammals including humans. Canadian Food Inspection Agency, Saskatoon, It is a zoonotic disease of particular concern in Canada, 3Canadian Wildlife Health Cooperative, rural areas where humans have close contact Department of Pathology and Microbiology, with domestic animals that can transmit the Atlantic Veterinary College, University of disease. Serological surveys on Toxoplasma Prince Edward Island, Charlottetown, Canada, gondii have been done in both humans and 4Department of Fisheries and Oceans Canada, animals in various parts of the world; however Winnipeg, Canada, 5Institute for Conservation in South Africa literature on this is either out- Research San Diego Zoo Global, Escondido, dated or scant. Therefore, a cross-sectional United States, 6Environment and Climate Change survey was conducted to investigate Toxoplasma Canada, Government of Canada, Edmonton, gondii seroprevalence and associated risk Canada factors in small ruminants, pigs, poultry and cats in the Oliver Reginald Tambo District in Toxoplasma gondii is an important zoonotic the Eastern Cape in South Africa between parasite that can be transmitted to humans June 2016 and October 2016. Household-level by contaminated food or water contamination and animal-level data were collected using a with feline feces containing oocysts, and close-ended questionnaire. The Toxoreagent, by consumption of raw or undercooked a latex agglutination test, was used for T. meat harboring the tissue cysts with the gondii antibody detection. Positive samples bradyzoites. Wildlife, including marine had agglutination patterns at dilutions of 1:64 mammals, are hunted and harvested by or greater, except for chickens, whose cut off different communities in the Canadian North, titre was 1:32. A household was classified as as part of traditions and culture, and as a Toxoplasma gondii seropositive if at least one critical food resource. In recent years, there species tested positive. The study revealed that has been increasing concern about high rates 125 / 150 farms (83.3%), 78 / 121 sheep (64.46%), of exposure to T. gondii in northern residents, 69 / 128 goats (53.91%), 36 / 106 pigs (33.96%), possibly through foodborne routes. The cycle 35 / 109 cats (32.11%) and 46 /137 chickens of the parasite is well known in terrestrial (33.58%) were seropositive for the parasite. ecosystems; however, the transmission Seropositivity was assessed for association dynamics between terrestrial felids and with potential risk factors. Among them, marine mammals is not well understood, age, municipality, climate, animal production especially in the Arctic where felids are rare system, rodent control, cat-feed access and cat above treeline. The aim of this study is to faecal disposal were found to be statistically determine the seroprevalence of antibody significant using the Chi-Squared test or odds to T. gondii in marine mammals frequently ratio, confirmed by the Fisher’s exact test. hunted by the northern communities. Serum The relatively high seroprevalence of T. gondii samples were obtained from grey seal detected in this study suggests that the infection (Halichoerus grypus), ringed seal (Pusa may pose a substantial public health risk through hispida), beluga whale (Delphinapterus the consumption of T. gondii infected raw meat leucas) and polar bear (Ursus maritimus) as well as contact with cat faeces. from different regions of the Canada. The

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 100 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

samples were tested by a commercial ELISA immitis, D. repens, and A. reconditum. Two kit and the Immunofluorescence Antibody dogs each (0.1%) were identified as PCR Test (IFAT) for detection of antibodies against positive for D. immitis and D. repens while 110 Toxoplasma gondii. We anticipate high dogs (7.7%) were identified as PCR positive for seroprevalence in polar bears as they could A. reconditum. None tested positive for more be infected through carnivory of terrestrial than one type of filarial parasite. D. immitis wildlife, as well as through waterborne incidence by antigen was 0.3%. Multiple oocysts, and that exposure to the parasite in logistic regression of A. reconditum PCR seals and beluga whales will be higher in more results identified province (p<.0001) and E. southern populations due to higher levels of canis serological status (p=.0011) as significant water contamination with the oocysts from predictors of A. reconditum infection. Risk felids. We will compare our results to the factors for positive A. reconditum PCR test published literature, and discuss challenges results included living in Salerno province (OR in serosurveillance for T. gondii in marine 4.51 vs. Avellino, 95% CI 2.44-8.32), living in mammals. Napoli province (OR 2.67 vs. Avellino, 95% CI 1.18-6.06), and seropositivity for E. canis (OR 2.71, 95% CI 1.49-4.92). Hunting dogs OA26 Canine Helminths II in this study were more likely to be PCR positive, and potentially microfilaremic, due July 9, 2019, 16:30 - 18:00 to A. reconditum than D. immitis or D. repens. Plenary Hall, Madison Ballroom (ABCD) This study identified a significant association between an infection transmitted by fleas, A. reconditum, and an infection transmitted by OA26.01 Molecular Detection of Filarial ticks, E. canis. These results emphasize the Parasites in Hunting Dogs From Southern Italy importance of broad-spectrum, year-round ectoparasite control to minimize vector- Melissa Beall1, Laura Pacifico2, Francesco mediated transmission of pathogens in hunting Buono2, Jennifer Braff1, Giovanni Sgori2, Jesse dogs from Southern Italy. Buch1, Benedetto Neola3, Diego Piantedosi2, Christian Leutenegger1, Ramaswamy Chandrashekar1 OA26.02 A Case of Canine Dracunculus Sp. 1IDEXX, Westbrook, United States, Infection from Spain 2Department of Veterinary Medicine and Animal Productions, University of Irina Diekmann1, Dr. Alaa Aldin Alnassan2, Dr. Naples Federico II, Naples, Italy, 3Istituto Majda Globokar2, Dr. Nikola Pantchev2, Dr. Lina Zooprofilattico Sperimentale del Mezzogiorno, Kurzrock2, Dr. Leticia Hernandez3, Dr. Javier Portici, Italy Lopez3, Dr. Ricardo Ruano4, Dr. Silvia Herrero4, M. De Vidales4, Prof. Dr. Georg von Samson- Several filarial parasites are known to infect Himmelstjerna1, Dr. Jürgen Krücken1 dogs including Dirofilaria immitis, D. repens, 1Institute for Parasitology and Tropical and Acanthocheilonema reconditum. While Veterinary Medicine, Freie Universität Berlin, microscopic exam of circulating microfilaria Berlin, Germany, 2IDEXX Laboratories, may allow differentiation based on Ludwigsburg, Germany, 3IDEXX Laboratories, morphological characteristics, such distinctions Spain, 4Clínica Mediterráneo, Madrid, Spain are not always possible in veterinary practice. This study investigated the prevalence of these The nematode genus Dracunculus contains parasites in hunting dogs from Southern Italy 14 accepted species, 9 of which parasitize using molecular methods. Blood was collected reptiles. Human dracuncolosis caused by from 1,433 dogs in Napoli, Avellino, and Salerno Dracunculus medinensis was an important provinces as part of a Campania region hunting neglected tropical disease but recent efforts dog health assistance program. Samples have eliminated the parasite from most were analyzed using SNAP® 4Dx® Plus (IDEXX endemic countries except of Chad, Ethiopia, Laboratories, Inc.) and real-time polymerase South Sudan and Mali. Shortly before its chain reaction (PCR) to amplify DNA from D. eradication, canine D. medinensis infections

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 101 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

have become considerably more frequent and Rhipicephalus sanguineus sensu lato (s.l), dogs now appear to be important reservoir the intermediate host and vector. Despite hosts maintaining the live cycle in endemic the world-wide distribution of R. sanguineus areas. However, Dracunculus lutrae (host: s.l., C. bainae had not been documented North American river otter Lontra canadensis) in the United States prior to 2017, when a and Dracunculus insigne (wide host range single case report of a dog in Florida with including raccoons, several mustelids and dermatitis was described. To determine the canids including dogs) infect predominantly prevalence of C. bainae in domestic dogs in North American carnivores. A 2-year-old, Oklahoma, client-owned and impounded dogs podenco-like (local Spanish breed) dog was were screened between January–October presented to a veterinarian in Toledo (Spain) 2018 for microscopic and molecular evidence due to an oozing ulcer at the hind left limb. The of infection. Dermal biopsies (6 mm) were dog had a history of abuse on a farm before processed via saline sedimentation; sediments being rescued about a year before attendance. were microscopically examined and, when From the ulcer, an approximately 12 cm microfilariae were identified, measurements anterior end of a nematode was extracted. The were taken. Microfilariae positive sediments dog had been regularly dewormed, carried and skin biopsy sections were tested by PCR a Seresto® collar and was under allopurinol to amplify filarioid DNA (12S and 5.8S rRNA therapy against leishmaniosis. Morphological gene fragments). Additionally, ticks infesting examination of the nematode excluded a surveyed dogs, including R. sanguineus s.l., filarioid species and Dracunculus sp. was were collected and screened for molecular suspected. Following PCRs and sequencing evidence of C. bainae infection. Microfilariae of partial cytochrome oxidase I (COI), 18S and were observed in 8.0% (20/250) of dogs, with 28S rRNA genes confirmed the allocation three dogs having organism consistent with to the genus Dracunculus but excluded D. C. bainae by morphometry. Cercopithifilaria medinensis, D. insignis and D. lutrae as well bainae DNA was amplified from skin biopsies as two undescribed genotypes previously of 4.7% (6/127) dogs, 2 of which were reported from L. canadensis as causative positive for C. bainae microfilariae on skin agents. Phylogenetic analysis using the COI sedimentation. Two dogs with microfilariae of sequence placed the specimen in a sister C. bainae were noted to have R. sanguineus position to D. insignis within the Dracunculus s.l. at the time of skin biopsy collection. To genus but outside of any established species date, C. bainae has not been detected in suggesting it represents a species not detected any ticks tested (n=112). The present study in carnivores before. To the knowledge of the documented C. bainae in Oklahoma dogs for authors, this is the first report of a Dracunculus the first time, with collective morphometric sp. infection of mammals from Europe. and molecular data indicating that the parasite is actively cycling within the state.

OA26.03 Detection and Prevalence of Cercopithifilaria Bainae Infection in Domestic OA26.04 The Impressive Spread of the Dogs and Ticks in Oklahoma Metastrongylid Heart and Lung Nematode Angiostrongylus Vasorum in Foxes As Megan Wohltjen1, Kellee Sundstrom1, Susan Cause for Increase of Cases of Canine Little1, Kelly Allen1 Angiostrongylosis 1Oklahoma State University, Stillwater, United States Dr. Nina Gillis-Germitsch1, Dr. Lucienne Tritten1, Cercopithifilaria bainae is a vector-borne Dr. Daniel Hegglin1, Prof. Peter Deplazes1, Prof. filarial nematode of dogs in areas of Europe, Manuela Schnyder1 Central America, and South America. The 1Institute of Parasitology, University Of Zurich, subcutaneous parasite is considered largely Zürich, Switzerland non-pathogenic, but occasional clinical signs include erythematous papules, polyarthritis, After its discovery in France around and pruritic dermatitis. Transmission to 1900, Angiostrongylus vasorum (“‘French canid hosts occurs via the blood-feeding of heartworm’) was first reported in dog

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 102 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

kennels in Switzerland and in Ireland in the OA26.05 Multiple Drug Resistance in 1960s, and later also in other European Ancylostoma Caninum: An Emerging Threat countries. A consistent increase of cases of to Canine Health canine angiostrongylosis, however, was only observed in the past decade. Pablo David Jimenez Castro1, Daniel Zuluaga2, Patricia Wetherly1, Razan Sheikh1, Russell To shed light on this dynamics, 3’954 blood Avramenko3, John Gilleard3, Ray Kaplan1 samples collected from Swiss foxes between 1University of Georgia, Athens, United States, 1986 and 2017 were tested for circulating A. 2University of Santa Rosa de Cabal, Santa vasorum antigen by ELISA. The samples were Rosa de Cabal, Colombia, 3University of allocated to 4 collection timeframes: 1986- Calgary, Calgary, Canada 1992, 1993-2002, 2003-2012 and 2013-2017. In north-eastern Switzerland, 1.9% (25/1343) and Over the past two years, we have become 1.7% (14/833) of foxes were antigen-positive aware of multiple cases of recurrent in the first two timeframes, respectively. hookworm infections in dogs due to Antigen-positivity increased in the following Ancylostoma caninum that are relatively two decades to 18.3% (17/93; 2003-2012) unresponsive to usual anthelmintic and to 62.0% (281/453; 2013-2017). In south- treatments. We now provide conclusive eastern Switzerland comparable prevalences evidence that the primary cause of these in foxes were determined for 1986-1992 recurrent hookworm cases is the development (7.9%; 13/164) and 1993-2002 (6.5%; 45/691).. of multi-dug resistance (MDR) in this parasite. Between 2003-2012, 38.5% (52/135) of foxes Three suspected-resistant and two susceptible were antigen-positive. In western Switzerland, isolates of A. caninum were established in 1.4% (3/222) of foxes were antigen positive laboratory dogs. In vitro bioassays including in the first but 53.3% (8/15) in the second the egg hatch assay (EHA) and the larval time frame. In parallel, necropsy of 467 development assay (LDA) were used to foxes around Zurich confirmed an increasing measure resistance to benzimidazoles and prevalence for A. vasorum from 20.5% (worm macrocyclic lactones, respectively, and fecal burden [WB]: 1-30, mean 7.3) in 2012 to 75.5% egg count reduction was measured following (WB: 1-50, mean 11.3) in 2017. successive treatment with fenbendazole, pyrantel and milbemycin oxime in two Our results indicate that A. vasorum laboratory dogs. In addition, samples of eggs prevalence in the Swiss fox population was and L3 were examined using deep-amplicon stable at low levels until 2002, whereas sequencing to evaluate the beta-tubulin it increased significantly since 2003 in all SNP frequency. The EHA IC50 value for our investigated areas. This corresponds to susceptible lab isolate was 0.25uM and for the time in which A. vasorum started to resistant isolates ranged from 2.79-3.73uM, be detected in dogs. We hypothesize that yielding resistance ratios (RR) of 11.1-14.9. LDA the increasing number of cases of canine IC50 value for the susceptible isolate was angiostrongylosis is due to a simultaneous 16.62nM and ranged from 91.53-1052nM for increase of A. vasorum prevalence in the resistant isolates, which yielded RR from 5.5- Swiss fox population, which constantly 63.3. Following treatment with fenbendazole, increased, particularly in urban areas. As is the percent reduction in faecal egg counts were case with the zoonotic cestode Echinococcus 63 and 84% (day 23), following treatment multilocularis, foxes play a crucial role as with pyrantel were 0 and 72% (day 10), and reservoir hosts. following treatment with milbemycin were 92 and 58% (day 14) for the resistant Tara and Worthy isolates, respectively. Beta-tubulin SNP were present only at codon 167; all but one sample from a resistant isolate yielded SNP frequencies greater than 88%. These in vitro, in vivo, and molecular data offer pivotal evidence that these A. caninum isolates are highly resistant to all anthelmintic classes

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 103 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

currently approved for use in dogs in the indicating that exposure through United States. Deep amplicon sequencing is a environmental contamination may occur in powerful new tool that can be used to study natural infection in dogs. As an exposure the molecular epidemiology of anthelmintic route, eating or licking grass or other plant resistance in A. caninum. material greatly increases the number of dogs at risk of infection for these parasites.

OA26.06 Infectivity of Shed Angiostrongylus Vasorum and Crenosoma Vulpis L3 to Dogs OA27 Epidemiology

William Robbins1, Dr. Spencer Greenwood1, Dr. July 9, 2019, 16:30 - 18:00 Laura Rogers2, Dr. Gary Conboy1 Breakout Room 2, Hall of Ideas E&H, Level 4 1Atlantic Veterinary College-UPEI, Charlottetown, Canada, 2Department of Fisheries and Land Resources, St. John’s, OA27.01 “Tekenscanner”: A Novel Canada Smartphone App for Companion Animal Owners and Veterinarians to Engage in Tick Metastrongyloid respiratory parasites, and Tick-Borne Pathogen Surveillance in the Angiostrongylus vasorum and Crenosoma Netherlands vulpis infect wild and domestic canids and Prof Frans Jongejan1, Suzanne de Jong1, are important pathogens in dogs. Recent 1 1 studies indicate gastropod intermediate Timo Voskuilen , Louise van den Heuvel , Rick Bouman1, Henk Heesen2, Carlijn IJzermans2, hosts infected with various metastrongyloids 1 spontaneously shed infective third-stage Laura Berger 1Utrecht Centre for Tick-borne Diseases, larvae (L3) into the environment via feces Utrecht University, Netherlands, 2Bayer Animal and mucus. Shed L3 retain motility up to Health, Mijdrecht, Netherlands 120 days but whether they retain infectivity remains unknown. To assess the infectivity of shed L3, the heart/lungs of 6 red fox The engagement of companion animal (Vulpes vulpes) were obtained from owners into the process of collecting trappers in Newfoundland, Canada. Lungs epidemiological data can be facilitated were examined for first-stage larvae (L1) through smartphone applications. Early 2018, by Baermann technique. A high number of the “tekenscanner“(Dutch for tick scanner) viable A. vasorum L1 and a small number of C. app was launched in the Netherlands with vulpis L1 were recovered from one fox; these the aim of engaging owners to record ticks were used to infect laboratory-raised Limax removed from their pets and submit these maximus. L3 recovered by artificial digestion ticks for identification and pathogen testing. were fed to two purpose-bred research A barcode was assigned to each tick for beagles (100 L3/dog). The L1 shed by these tracking of each submission to our laboratory 2 dogs were used to infect 546 L. maximus for taxonomic identification. DNA extracted (2,000 - 10,000 L1/slug). Shedding of L3 was was PCR amplified, subjected to reverse line induced by anesthetizing slugs in soda water blot hybridization (RLB) and screened for and transferring them into warm (45° C) tap pathogens. Results were added to the same water for at least 8 hrs. Recovered shed L3 app within two weeks. were aliquoted on romaine lettuce in 6-well tissue culture plates (80-500 L3/well) and The app was downloaded 5591 times and kept at 16° C/75% Relative Humidity. Four resulted in the collection of 1005 ticks from research beagles were exposed to 100 L3/dog 406 dogs and 268 ticks from 111 cats. Ixodes from the larvae held at 0, 2, 4 or 8 weeks after ricinus was the predominant species (90.0%), shedding. All four dogs began shedding C. with all stages found on dogs as well as vulpis L1 by 26-36 days post-infection (PI). All on cats. Ixodes hexagonus (7.3%) female four dogs began shedding A. vasorum L1 by and nymphal ticks were also identified on 50 days PI. Infectivity of L3 for the definitive both hosts, whereas adults of Dermacentor host was retained in both metastrongyloids reticulatus (2.4%) and Rhipicephalus

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 104 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

sanguineus (0.2%) were exclusively found on delayed anaphylactic reaction to mammalian dogs. Nearly 15% of the ticks recovered from meat in human patients previously exposed to dogs carried one or more pathogens, whereas bites of certain tick species. In dogs, alpha-Gal 13.8% of the ticks removed from cats were is a self-antigen and their immune system is infected. Ixodes ricinus collected from dogs thus not expected to naturally produce Abs contained Borrelia spp. (1.9%), Babesia spp. toward the glycan molecule. However, results (0.7%), Anaplasma phagocytophilum (1.3%), of this preliminary study demonstrate the Neoehrlichia mikurensis (2.9%) and Rickettsia occurrence of canine IgG, IgM and IgE serum helvetica (7.3%). Ixodes ricinus recovered Abs against alpha-Gal for the first time, and from cats were infected with Borrelia spp. suggests that a tick bite can sensitize dogs (1.9%), Babesia spp. (0.4%), Anaplasma to alpha-Gal and potentially trigger delayed phagocytophilum (1.9%), Neoehrlichia allergic reaction to mammalian meat following mikurensis (2.6%) and Rickettsia helvetica the induction of specific IgE. The results also (6.7%). Three Rhipicephalus sanguineus, on suggest the protective role of anti-alpha-Gal dogs from France and the USA imported into Abs against pathogens transmitted by ticks. the Netherlands, were negative. Our findings open a completely new scientific perspective, which may contribute to a better The tekenscanner app is a versatile tool to understanding of the mechanisms involved in use for submission of ticks and facilitated the pathogenesis of this unique food allergy, fast feedback of test results. Community and highlight the possibility that anti-alpha- engagement through the app is suitable for Gal Abs may help to prevent canine vector- identifying hot spots for ticks and pathogens borne diseases. and provided an early warning system for exotic ticks invading the Netherlands. OA27.03 Diversity of Parasites of Companion Dogs and Cats in East and Southeast Asia. OA27.02 Detection of Anti-Alpha-Gal Part 1: Endoparasites Antibodies in Dogs and the Possible Implications in Red Meat Allergy and Dr. Vito Colella1, Mr. Do Yew Tan2, Mrs. Na Protection to Infectious Diseases Lu3, D.r Fang Fang4, Yin Zhijuan4, Jiangwei Wang5, Xin Liu6, Junyan Dong7, Wisnu Dr. Adnan Hodžić1, Dr. Lourdes Mateos- Nurcahyo8, Upik Kesumawati9, Virginia Hernández2, Dr. Michael Leschnik3, Dr. Pilar Venturina10, Kenneth Boon Yew Tong11, Yi- Alberdi2, Dr. Ryan O. M. Rego4, Dr. Marinela Lun Tsai12, Piyanan Taweethavonsawat13, Contreras2, Dr. Margarita Villar2, Prof. José de Saruda Tiwananthagorn14, Thy Anh Dang15, la Fuente2, Dr. Alejandro Cabezas-Cruz5, Dr. Thong Quang Le15, Khanh Linh Bui16, Malaika Georg Duscher1 Watanabe17, Puteri Azaziah Megat Abd Rani17, 1Institute of Parasitology, Department of Giada Annoscia1, Viet Linh Nguyen1, Frederic Pathobiology, University of Veterinary Beugnet18, Domenico Otranto1, Lénaïg Halos18 Medicine Vienna, Vienna, Austria, 2SaBio, 1University Of Bari, Bari, Italy, 2Boehringer Instituto de Investigación de Recursos Ingelheim Animal Health, Singapore, Cinegéticos, IREC-CSIC-UCLM-JCCM, Ciudad Singapore, 3Boehringer Ingelheim Animal Real, Spain, 3Small Animal Clinic, Department Health, Beijing, China, 4College of Animal for Companion Animals, University of Science and Technology, Guangxi University, Veterinary Medicine Vienna, Vienna, Austria, China, 5Aibi-pet chain pet Veterinary clinics 4Institute of Parasitology, Biology Center, , Shanghai, China, 6Meilian chain Veterinary Czech Academy of Sciences, eské Budějovice, clinics, Beijing, China, 7Research pet hospital Czech Republic, 5UMR BIPAR, INRA, ANSES, of Ministry of Public security of China, Nanjing, Ecole Nationale Vétérinaire d’Alfort, Université China, 8Faculty of Veterinary Medicine, Gadjah Paris-Est, Maisons-Alfort, France Mada University, Yogyakarta, Indonesia, 9Faculty of Veterinary Medicine, Bogor Specific IgE antibodies (Abs) against the University Indonesia, Jakarta, Indonesia, carbohydrate galactose-alpha-1,3-galactose 10College of Veterinary Science and Medicine, (alpha-Gal) has recently been linked to Central Luzon State University, Nueva

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 105 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Ecija, Philippines, 11AAVC - Animal & Avian this geographical area as highly endemic for Veterinary Clinic, Singapore, Singapore, these pathogens. Coccidia, Strongyloides 12Department of Veterinary Medicine, spp., Trichuris spp., Trichomonas spp., National Pingtung University of Science and Dyphillobothriidae, Dipylidium caninum Technology, Pingtung, Taiwan, 13Faculty of and trematodes were also reported in both Veterinary Science, Chualalongkorn University, dogs and cats. Peaks in the occurrence of Bangkok, Thailand, 14Faculty of Veterinary endoparasites were recorded in selected Medicine, Chiang Mai University, Chiang Mai, countries such as Philippines and Vietnam Thailand, 15Faculty of Animal Science and with respectively 32.5% and 23% of the pets Veterinary Medicine, Nong Lam University, enrolled diagnosed with these pathogens. Ho Chi Minh, Vietnam, 16Vietnam National University of Agriculture, Hanoi, Vietnam, 17University of Putra, Petaling Jaya, Malaysia, OA27.04 Diversity of Parasites of Companion 18Boehringer Ingelheim Animal Health, Lyon, Dogs and Cats in East and Southeast Asia. France Part 2: Ectoparasites and Vector-Borne Diseases Asia is the largest continent in the world with an increasing number of dogs and cats Vito Colella1, Do Yew Tan2, Na Lu3, Fang Fang4, kept as companion animals. Nonetheless, Yin Zhijuan4, Jiangwei Wang5, Xin Liu6, Junyan in most of Asian countries, there is a lack Dong7, Wisnu Nurcahyo8, Upik Kesumawati9, of knowledge on the etiological agents Virginia Venturina10, Kenneth Boon Yew Tong11, parasitizing dogs and cats, including those Yi-Lun Tsai12, Piyanan Taweethavonsawat13, transmissible to humans. Saruda Tiwananthagorn14, Thy Anh Dang15, Thong Quang Le15, Khanh Linh Bui16, Malaika A collaborative field study involving 16 Watanabe17, Puteri Azaziah Megat Abd Rani17, veterinary institutions in 8 regions of East Giada Annoscia1, Viet Linh Nguyen1, Frédéric Asia (China, Taiwan) and Southeast Asia (SEA; Beugnet18, Domenico Otranto1, Lénaïg Halos18 Indonesia, Malaysia, Philippines, Singapore, 1University Of Bari, Bari, Italy, 2Boehringer Thailand, Vietnam) was set up to assess the Ingelheim Animal Health, Singapore, occurrence and prevalence of canine and Singapore, 3Boehringer Ingelheim Animal feline parasites. Health, Beijing, China, 4College of Animal Science and Technology, Guangxi University, From June 2017 to June 2018, a total of 2382 Guangxi, China, 5Aibi-pet chain pet Veterinary client-owned animals (1229 dogs and 1153 cats) clinics, Shanghai , China, 6Meilian chain were enrolled by veterinary facilities in each Veterinary clinics, Beijing, China, 7Research country. All animals had a history of regular pet hospital of Ministry of Public security of outdoor access, without any parasiticide China, Nanjing, China, 8Faculty of Veterinary treatment in the previous months. Animals were Medicine, Gadjah Mada University, Yogyakarta, identified and their lifestyle and anamnestic Indonesia, 9Faculty of Veterinary Medicine, data recorded, along with a complete Bogor University Indonesia, Jakarta, Indonesia, veterinary examination including parasitological 10College of Veterinary Science and Medicine, investigations for the diagnosis of both ecto- Central Luzon State University Nueva Ecija, and endoparasites. In particular, the occurrence Nueva Ecija, Philippines, 11Animal & Avian of endoparasites was assessed via qualitative Veterinary Clinic, Singapore, Singapore, coprological tests (flotation, sedimentation 12Department of Veterinary Medicine, and Baermann-Wetzel techniques). National Pingtung University of Science and Endoparasites were identified in 12.6% and Technology, Pingtung, Taiwan, 13Faculty of 13.9% of dogs and cats. Nematodes of the Veterinary Science, Chualalongkorn University, families Ancylostomatidae (8.9% and 6.4%) and Bangkok, Thailand, 14Faculty of Veterinary Toxocaridae (2.5% and 3.9%) were the most Medicine, Chiang Mai University, Chiang Mai, frequent gastrointestinal parasites detected. Thailand, 15Faculty of Animal Science and Veterinary Medicine, Nong Lam University, Ancylostomatidae were diagnosed in 16% of Ho Chi Minh, Vietnam, 16Vietnam National dogs and 12% of cats from SEA, indicating University of Agriculture, Hanoi, Vietnam,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 106 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

17University of Putra, Petaling Jaya, Vietnam, Acanthocheilonema reconditum 22.1%, A. 18Boehringer Ingelheim Animal Health, Lyon, platys 20.2%, Mycoplasma haematoparvum France 14.7%, Babesia vogeli 5.5%, Babesia spp. 4.3%, and Hepatozoon canis 3.1%. Fecal The 2382 animals enrolled as previously samples from 154 dogs were assessed using described were examined for ectoparasites centrifuged fecal floatation and fecal antigen and vector-borne pathogens. The body testing (Fecal Dx® , IDEXX Laboratories) of surface of each animal was examined for the which 72.1% were infected ≥1 parasite and presence of ectoparasites (ticks, fleas, lice, of these, 21.6% harbored ≥2 species. More and mites), including a thorough examination samples were positive for hookworms on underneath the third eyelid to detect Thelazia fecal antigen (68.8%) versus fecal floatation callipaeda eyeworms. A deep skin scraping (58.45%). There was whipworm antigen and/or a scotch test was performed only in detected in 14.9% of samples and positive case of lesions evocative of sarcoptic mange/ ova identification in 18.8%. The incidence of demodicosis or cheyletiellosis, respectively, roundworms was 1.9% on both tests. Based and ear wax examined for the presence of on the results of this study, dogs in Grenada Otodectes cynotis. are at significant risk for parasitic and vector-borne disease, several of which have zoonotic potential. This is the first report OA27.05 Finding What You Aren’t Looking of Mycoplasma spp. infection in Grenadian For: A Survey of Parasitic and Infectious dogs and highlights the importance for Diseases in Dogs in Grenada, West Indies veterinarians to be aware of the infectious organisms threatening their patients. The Dr. Tara Paterson1, Dr. Marta Lanza Perea1, Dr. veterinarian cannot find the underlying Jancy Hanscom2, Dr. Phyllis Tyrrell2, Dr. Dave etiology of disease if one is not looking for it. Elsemore2, Dr. Christian Leutenegger2, Dr. Ramaswamy Chandrashekar2 To assess the exposure to vector-borne 1St. George’s University, School of Veterinary pathogens in dogs, blood samples were Medicine, St. George’s, Grenada, 2IDEXX tested for Dirofilaria immitis, Ehrlichia spp., Laboratories, Inc., Westbrook, United States Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania spp. with SNAP® 4Dx Grenada’s tropical climate and prevalence Plus and SNAP® Leishmania tests. In cats the of vectors pose significant risk of infectious SNAP® Feline Heartworm Antigen Test Kit disease transmission in animals and people. was used to detect D. immitis. Blood samples Preventive health care is uncommon in were also stored on Whatman® FTA® cards Grenada due to economic limitations and for molecular detection of DNA from filarial poor understanding of canine diseases, parasites, and for species of the genera leaving dogs vulnerable to infection. Since Leishmania, Babesia and Hepatozoon. heartworm, ehrlichiosis and anaplasmosis are Ectoparasite infestations were reported endemic in Grenada, often other differential in 30.4 and 27.5% of dogs and cats, diagnoses are overlooked when faced respectively. Diversity of flea species included with a sick canine patient. The aim of this Ctenocephalides felis and C. orientis and study was to survey a population of dogs for the ticks Rhipicephalus sanguineus, R. presenting to the St. George’s University haemaphysaloides and R. turanicus. Exposure Junior Surgery & Anesthesia Lab using a to tick-borne pathogens was reported in 18.5% comprehensive diagnostic approach. Based of dogs (14.5% Ehrlichia and 7% Anaplasma), on IDEXX SNAP® 4Dx® Plus test results from with 16% and 10% of dogs from Philippines 166 dogs, 14.5%, 39.2%, 22.9% and 0% of and Taiwan, respectively, scoring positive for dogs were positive for Dirofilaria immitis, D. immitis. To our knowledge, we reported for Ehrlichia canis, Anaplasma platys, and Borrelia the first time the occurrence of Leishmania burgdorferi, respectively. Comprehensive spp. in dogs from Philippines and Vietnam, infectious disease PCR analysis performed and of D. immitis in a cat from Indonesia. on 163 samples identified the following: E. This multicenter study represents a canis 23.95%, Mycoplasma haemocanis 22.7%, cornerstone in the knowledge of the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 107 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

occurrence of parasites infecting dogs and OA28 Alternative treatments for cats kept as companion animals in East Parasites in Ruminants I and Southeast Asia, including species of zoonotic concern, ultimately leading to July 9, 2019, 16:30 - 18:00 the establishment of effective treatment Breakout Room 3, Hall of Ideas F&I, Level 4 strategies and refined recommendations for veterinarians and pet owners. OA28.01 The Nordic Seaweeds Saccharina Latissima and Laminaria Digitata Have OA27.06 Prevalence of Trypanosomosis Potent Anthelmintic Effects In Vitro (Trypanosoma Evansi) of Dromedaries in the Sahel of Tunisia Charlotte Smith Bonde1, Louis Bornancin2, Associate Professor Andrew Richard Williams1, Pr. Mohamed Habib Jemli1, Dr. Henda Associate Professor Helena Mejer1, Associate Boulajfen2, Dr. Ahmed Lahouar1, Dr. Talel Professor Henrik Toft Simonsen2, Dr. María Hamza3, Dr. Haykel Kessa4 Martínez Valladares3, Professor Stig Milan 1National School of Veterinary Medicine, Sidi Thamsborg1 Thabet, Tunisia, 2FAO, Tunisia, 3CRDA, Tunisia, 1University Of Copenhagen, Dept. of Veterinary 4CRDA, Tunisia and Animal Sciences, Frederiksberg, Denmark, 2Technical University of Denmark, Dept. of An epidemiological study was conducted Biotechnology and Biomedicine, Kongens between 2013 and 2016 in central Tunisia to Lyngby, Denmark, 3CSIC-Universidad de León, evaluate the prevalence of Trypanosoma evansi Dept. of Animal Health, Grulleros, Spain infection in camels by the direct agglutination test (CATT Test®). A total of 1076 dromedaries Seaweed contains an abundance of bioactive of both sexes were examined (96 males compounds, and some seaweed species and 980 females), aged from 1 to 18 years, have been used as natural deworming belonging to 3 regions of the Sahel of Tunisia agents for centuries in traditional Chinese were selected for the survey. medicine. In this study, we investigated the in vitro anthelmintic (AH) activity of 262 animals were seropositive (24.3%) with extracts of seaweed, from Nordic waters. a net prevalence of the disease in autumn We prepared three different extracts: (37%) and in animals over 4 years (89%). The hexane, dichlormethan:methanol (DCM), Monastir region was the most affected (42%). and water:methanol (WM), from four Appropriate treatment (Cymelarsan®) of all seaweed species: Saccharina latissima, seropositive patients resulted in a significant Laminaria digitata, Ascophyllum nodosum, decrease of clinical cases for at least two and Palmaria palmata. The AH activity was years in controlled flocks. The clinical signs of assessed using an Ascaris suum third stage high suspicion of trypanosomosis observed larvae (L3) mortality assay (1 mg DM/mL in dromedaries were weight loss, anemia, dissolved in DMSO). Moving or coiled-up edema in the sloping regions and sometimes larvae were counted as alive, and immobile locomotors disorders and abortions in females. or straight larvae as dead. Extracts with more apolar compounds (hexane, DCM) showed higher AH activity than extracts with polar compounds (WM), and the most potent extracts originated from S. latissima and L. digitata, with an average mortality of >95% after 48 hours. Extracts from A. nodosum had significantly lower AH effect after 48 hours. The extracts were further tested for AH effect in vitro against Teladorsagia circumcincta L1 (1 mg/mL, 48 h). The results showed a high AH activity of S. latissima and very low activity of A. nodosum extracts. An egg hatch assay

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 108 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

(EHA), performed using the same extracts widely used as bio-pesticides and are part (1 mg/mL, 48 h) and T. circumcincta eggs, of our diet. The crystal protein, Cry5B (FDA only showed >95% inhibition by the polar S. approved orphan drug status), from Bacillus latissima extract. We conclude that the Nordic thuringiensis (Bt) was earlier shown to be seaweeds Saccharina latissima and Laminaria efficacious against Ancylostoma ceylanicum digitata have strong in vitro AH effects (hookworm) and Ascaris suum (roundworm). against common pig and sheep nematodes, However, in these experiments spore-crystal and the AH activity is caused mainly by apolar lysates were used to test toxicity to GINs. compounds. Bio-guided fractionation will be To avoid the dissemination of live spores used to identify the active compounds, and and the potential enterotoxicity with Bt, we feeding trials of infected livestock will reveal genetically engineered Bt to form a non- whether seaweeds as a bioactive forage can sporulating Bacillus. After an inactivation play a role in future nematode control. step, the Inactivated Bacillus with Cytosolic Crystal (IBaCC) remained highly toxic to hookworms both in vitro and in a hamster OA28.02 Microbial Crystals for the Treatment infection model. IBaCC significantly reduced of Veterinary Gastrointestinal Nematodes Ascaris worm burden in infected pigs. To isolate Purified Cry5B Crystals (PCC) for Ambily Abraham1, Yan Hu1, David Gazzola1, pharmaceutical applications, we devised a Hanchen Li1, Kelly Flanagan1, Tasia Kellogg1, two-step scalable process that generated > Mostafa Elfawal1, Martin K. Nielsen2, Anne 90% pure Cry5B protein crystals that was Zajac3, Joseph F. Urban4, Katherine Petersson5, able to cure hookworm infection in hamsters. Jennifer Ketzis6, Gary Ostroff1, Raffi Aroian1 Additionally, within days of treatment these 1UMass Medical School, Worcester, United crystals lowered the Parascaris equorum fecal States, 2M.H. Gluck Equine Research Center, egg count in horses. In a pilot study of African Department of Veterinary Science, University Green Monkeys administered with Cry5B, of Kentucky Lexington, KY, United States, a decrease in egg and larvae shedding of Lexington, United States, 3Dept. Biomedical Strongyloides (threadworms) was observed. Sciences and Pathobiology, VA-MD College of Taken together, we have developed two drug Veterinary Medicine, Virginia Tec, Blacksburg, forms - an encapsulated, inactivated bacteria United States, 4USDA, Agricultural Research (IBaCC) and purified crystals (PCC) to meet Service, Beltsville Agricultural Research the ongoing challenge of GIN parasitic Center, Animal Parasitic Diseases Laboratory, infections. Beltsville, United States, 5Dept. Fisheries, Animal & Veterinary Science, University of Rhode Island, Kingston, United States, 6Ross OA28.03 Anthelmintic and Metabolomic University School of Veterinary Medicine, St. Analyses of Chicory (Cichorium Intybus) Kitts, West Indies Extracts Revealed an Industrial By-Product With Activity Against Drug-Sensitive and Gastrointestinal nematode (GIN) parasites Drug-Resistant Nematodes are one of the leading threats to the animal industry, resulting in decline in reproductive Dr. Miguel Peña-Espinoza1, Dr. Louis performance, immunity and sometimes death Bornancin2, Dr. Andrew R. Williams3, Dr. of the animal, amounting to huge economic Henrik T. Simonsen2, Prof. Stig M. Thamsborg3, losses. With the ever-increasing consumer Dr. Rodrigo Lopez-Muñoz1 demand for chemical-free animal products 1Universidad Austral de Chile, Valdivia, Chile, and the rise of anthelmintic resistance 2Technical University of Denmark, Kongens against current small molecule drugs in these Lyngby, Denmark, 3University of Copenhagen, parasites, a need for a new inexpensive and Frederiksberg, Denmark scalable treatment with minimal side effects is on the surge. We have focused on utilizing Chicory (Cichorium intybus) is a bioactive invertebrate-specific bacterial pore-forming crop rich in sesquiterpene lactones (SL) with crystal proteins that have evolved naturally reported anthelmintic activity in livestock. to disrupt host membranes, have been Evaluation of SL-extracts from different

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 109 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

chicory materials can help identifying the OA28.04 Scientific Validation of main antiparasitic compound(s), leading to Ethnobotanicals Used Against Rhipicephalus selection of crops with increased activity and/ Microplus in Pakistan or the isolation of novel parasiticides. Here, we evaluated the anthelmintic activity and Dr. Muhammad Arfan Zaman1, Professor metabolomic profiling of SL-extracts from Zafar Iqbal2, Dr. Rao Zahid Abbas2, Professor different chicory materials against drug- Muhammad Fiaz Qamar1 sensitive and drug-resistant nematodes. 1College of Veterinary and Animal Sceinces, Jhang, Pakistan, 2University of Agriculture, Fresh leaves from forage chicory cultivated in Faisalabad, Pakistan Denmark (var. Spadona, sampled in 2013, 2015 and 2016) and Chile (var. Choice, sampled in Aims 2016) were used for SL extraction. SL were The objective of the current study was also extracted from root chicory pulp, a by- to evaluate the anti-tick, activities of a product of the industrial extraction of inulin herbal formulation based on leaves of from chicory roots (var. Sativum). The five SL- Azadirachta indica and Nicotiana tabacum, extracts were tested against a Caenorhabditis flowers of Calotropis procera and seeds of elegans drug-sensitive strain, and the two Trachyspermum ammi. most active extracts were evaluated against an ivermectin (IVM)-resistant C. elegans Materials & methods strain. The most potent SL-extract was further Rhipicephalus (Boophilus) microplus ticks tested against Ascaris suum third-stage larvae were used for evaluation of acaricidal activity. (L3). All SL-extracts were analysed by ultra- The adult immersion test (AIT) (Sabatini high-performance liquid chromatography and et al., 2001) and larval packet test (LPT) metabolomic analyses were performed using (Luguru et al., 1984) were carried out for in the Global Natural Product Social Molecular vitro acaricidal activity of extract. For in vivo Networking database. evaluation of acaricidal activity, 15, 30 and 45% concentrations of extract were prepared Four SL-extracts induced a dose-dependent in distilled deionized water. inhibition of worm motility in drug-sensitive C. elegans. Root pulp and Spadona-2015 extracts Results were the most active SL-extracts (EC Root The formulation demonstrated anti-tick pulp: 72.1 g/mL vs Spadona-2015: 379 g/ activity by inhibiting the egg laying (Adult mL; P<0.0001). Only the root pulp extract₅₀ immersion test), larval mortality (Larval significantlyμ reduced the motility of IVM-μ packet test) and reduced tick intensity/ resistant C. elegans (P<0.001). Furthermore, infestation on animals. A dose dependent the root pulp extract induced a marked anti-tick effect was observed in all the and dose-dependent inhibition of A. suum tests carried out in this study. Egg laying L3 motility (EC : 118 g/mL). Metabolomic was significantly lower in ticks exposed to analyses confirmed the presence of SL in all different concentrations of herbal extract tested extracts ₅₀and a μdistinct SL profile in the compared with those exposed to distilled root pulp extract. water used as control. Likewise, there was a significant reduction (p<0.05) in the number In conclusion, root chicory pulp showed potent of ticks exposed to 45% herbal extract anthelmintic activity against drug-sensitive and compared with control. drug-resistant nematodes and further research should explore its potential as an antiparasitic Conclusion by-product. Undergoing bioactivity-based Pakistan is rich in indigenous knowledge, networking analyses could reveal the main diversity of plants and their usage, in anthelmintic compound(s) in chicory. traditional veterinary medicine. The herbal formulation demonstrated anti-tick activity by inhibiting the egg laying, larval mortality and reduced tick intensity/infestation on animals. The herbal formulation is suitable for the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 110 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

resource-poor farmers as a broad spectrum immunogenic epitopes among TvCSP genes. antiparasitic. The contents of the formulation Our analysis revealed that a novel family are cheap, commonly available, and easy to of T. vivax transmembrane proteins, which use as a decoction. Moreover, it is not a new we called vivaxin, are consistently the most animal husbandry input as farmers are already immunogenic non-VSG antigens in natural using these plants individually in animal health T. vivax infections across the Africa and and production. South America. The vivaxin gene family consists of 44 paralogs, divided into 14 clades, displaying stable polymorphism OA29 Unusual Protozoa and constitutive expression in bloodstream infections. Immuno-fluorescent microscopy July 9, 2019, 16:30 - 18:00 confirms that vivaxin proteins localize to the Breakout Room 4, Hall of Ideas G&J, Level 4 entire cell surface. Several vivaxin proteins were expressed recombinantly and used to vaccinate mice; this showed that specific OA29.01 Characterization of Vivaxin, a family members produce high antibody Novel Bloodstream-Stage, Species-Specific, titres. Currently, we are conducting challenge Cell-Surface Family As Potential Vaccine experiments in goat to examine the ability Candidates Against the Livestock Parasite of these recombinant antigens to induce Trypanosoma Vivax protective immunity. This is the first report of a non-VSG TvCSP family being vaccine Alessandra Romero-Ramirez1, Dr. Delphine candidates against AAT. The characterization Autheman2, Dr. Sara Silva Pereira3, Dr. Marta of these immunogenic but invariant surface M. Geraldes Teixeira4, Dr. Gavin Wright2, Dr. proteins will facilitate further study of their Rosangela Zacarias Machado5, Dr. Andrew P. biological functions, and represents a new Jackson1 approach to preventing AAT, a goal that 1Department of Infection Biology, Institute would bring profound benefits to animal of Infection and Global Health, University of health and productivity. Liverpool, Liverpool, United Kingdom, 2Cell Surface Signalling Laboratory, Wellcome Trust Sanger Institute, Hinxton, United Kingdom, OA29.02 Prevalence and Molecular 3Instituto de Medicina Molecular–João Lobo Identification of Babesia Species in Antunes, Faculdade de Medicina,Universidade Asymptomatic Dogs (Canis Familiaris) in the de Lisboa, Lisboa, Portugal, 4Institute of Federal Capital Territory, Abuja, Nigeria Biomedical Sciences, University of São Paulo (ICB/USP), São Paulo, Brazil, 5Department Dr. Sylvester Obeta1, Prof. Idris Lawal2, Prof. of Veterinary Pathology, School of Agrarian Joseph Natala2, Prof. Olorunfemi Jegede1, and Veterinary Sciences (FCAV/UNESP), Prof. Maxwell Opara1, Dr Ndudim Ogo3, Dr Jaboticabal, Brazil Emmanuel Balogun2,4 1University Of Abuja, Gwagwalada, Nigeria, Trypanosoma vivax is a major animal 2Ahmadu Bello University, Zaria, Zaria, Nigeria, pathogen causing African Animal 3National Veterinary Research Institute, Vom, Trypanosomiasis (AAT) in livestock across Nigeria, 4University of Tokyo, Hongo, Japan Africa and South America. No vaccine is available for AAT since antigenic variation Canine babesiosis is a protozoan tick-borne of the Variant Surface Glycoprotein (VSG) disease caused mainly by several species coat displayed on the parasite surface leads of Babesia with worldwide distribution. to effective immune evasion. However, the Accurate identification of the parasite species T. vivax genome contains species-specific, is important to the monitoring, management cell -surface genes (TvCSP) expressed and control of the disease. The aim of during the bloodstream stage, which we this study therefore was to determine the believe provide non-VSG and antigenically prevalence of Babesia infections in dogs invariant targets for vaccination. We used a as well as identify the species genotype in customized peptide microarray to identify the Federal Capital Territory, (FCT) Abuja

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 111 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

– Nigeria. Peripheral blood samples were in vitro. TrxR had a high catalytic activity to collected from 480 asymptomatic dogs in the the substrate and this activity was strongly six Area Councils (Abaji, Bwari, Gwagwalada, dependent on TrxR concentration. Using Municipal, Kwali and Kuje) via anti-rabies a combination of homology modeling and vaccination campaign and assayed with a light domain prediction methods, data suggested microscope using Geimsa stain. Genomic DNA that the electron-accepting site of TrxR is was extracted from whole blood which tested the Cys105-VNVGCys110 motif located at positive for Babesia spp., using DNA extraction the N-terminus and the electron transfer site kits. DNA amplification of the 18S ribosomal of TrxR is the Cys547-XXXX-Cys552 motif RNA gene was employed using genus specific located at the C-terminus. An inhibitor library primers. The amplicons were electrophoresed was constructed and tested, and several in 1.5 agarose gel, purified, and then directly inhibitors were then selected. To reveal the sequenced using Big Dye Terminator Cycle action sites of different inhibitors, a mutant Sequencing Kit. The BLAST search analysis pool was constructed and expressed as His- was conducted in NCBI database, and the tagged fusion proteins in Escherichia coli, sequences compared with reference data in according to the manufacturer’s instructions. the GenBank. The prevalence of Babesia spp. The reducing activity of the N-terminal infection by thin blood smears microscopy mutants Bmi TrxR-C105, Bmi TrxR-C110, and showed presence of intraerythrocytic Bmi TrxR-C105-C110 disappeared on the merozoites, in 3.1% (480/15) of the dogs DTNB and thioredoxin (Trx) substrates. The sampled. The PCR assay showed 53.3% (15/8) C-terminal mutants Bmi TrxR-C547, Bmi amplification of Babesia sp. at 612bp gene TrxR-C552, and Bmi TrxR-C547-C552 all segments on gel electrophorogram. BLAST reacted with DTNB; however, His-tags affected search analysis of the parasite sequences the reaction with Trx. The inhibition efficiency showed 100% sequence similarity with Babesia of 1-chloro-2, 4-dinitrobenzene (CDNB) on canis vogeli in the GenBank. This study mutant C547 decreased, and the inhibition documents B. canis vogeli as the species of CDNB was attenuated with the cysteine present in the sampled dog population and to mutation at position 547, indicating that the the best of our knowledge, the first molecular target of CDNB is 547 cysteine. These results identification of the Babesia sp. in the FCT, indicate that the site of electron acceptance Nigeria. Further studies need to be directed in the N-terminal plays the same key roles towards utilizing the PCR protocol to confirm as other homologs and the site of electron parasite species in tick vectors from the region. transfer in the C-terminal can be affected by the C-terminal His-tag. In conclusion, TrxR is associated with the resistance to oxidative OA29.03 Effects of a TrxR Amino Acid stress and is a promising target for Babesia Mutation on Enzyme Activity in Babesia microti therapy. Microti and the Different Roles of Inhibitors

Houshuang Zhang1, Jinmiao Lu1, Jie Cao1, Yongzhi Zhou1, Haiyan Gong1, Jinlin Zhou1 1Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China

Babesia microti is a zoonotic pathogen, which mainly parasitizes mammalian erythrocytes. The oxidative stress induced by the aerobic environment and immune system causes damage from ROS in the parasite. However, little is known regarding anti-oxidation strategies in Babesia microti. In the current study, the expression of TrxR was positively correlated with the ROS level both in vivo and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 112 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA29.04 Identification of East Coast eastern and southern Africa. The finding may Fever and Corridor Disease Discriminatory have implications on the use of recombinant Sequence Variations in Allele 1 of the p67 subunit vaccine against a challenge with Sporozoite Antigen Gene p67 buffalo-derived T. parva.

Dr. Donald Mukolwe Lubembe1,2, Dr. David O Odongo3, Dr. Sibeko P Kgomotso1 OA29.05 Differential Gene Expression 1Vectors and Vector-borne Disease Research Response in Domestic Cats (Felis catus) Programme, Department of Veterinary Experimentally Infected with Cytauxzoon felis Tropical Diseases, University of Pretoria, Onderstepoort 0110, South Africa., 2Faculty Dr. Ruth Scimeca1, Dr Mason Reichard1 of Veterinary Medicine and Surgery, Egerton 1Oklahoma Sate University, Stillwater, United University, Kenya., 3School of Biological States Sciences, University of Nairobi, Kenya Cytauxzoonosis is a fatal tick transmitted East Coast fever (ECF) and Corridor disease protozoan disease of domestic cats. No study (CD) caused by the cattle- and buffalo-derived has profiled the gene expression response Theileria parva respectively are considered to of cats to infection with Cytauxzoon felis. be the most economically important disease The aim of our work was to determine syndromes of cattle theileriosis in Africa. the host-parasite immune transcriptional The gene encoding the T. parva sporozoite elements related with clinical disease in cats surface antigen, p67, has been explored for the experimentally infected with C. felis. True development of a recombinant vaccine. Similar Single Molecule Sequencing (tSMS) was used to the current live vaccine, the major limitation to analyze the whole genome of acutely and of the p67-based recombinant vaccine is the chronically infected C. felis cats. Amblyomma heterogeneity of T. parva field isolates. Four americanum nymphs were acquisition fed on p67 alleles have been reported, where allele two cytauxzoonosis survivor cats. Replete 1 and alleles 2,3,4 have been associated with A. americanum nymphs were collected and ECF and CD respectively. However, allele 1 stored in controlled temperature and humidity has also been identified in buffalo-derived T. chambers until they molted to adults. Adult parva in South Africa; it is not clear if parasites A. americanum were then transmission fed possessing this allele can potentially cause on three naive cats. Controls samples were the re-emergence of ECF in this country. We collected before infestation and infection with assessed the diversity of allele 1 in buffalo- C. felis. tSMS results showed a total of 1068 and cattle-derived T. parva strains from differentially expressed genes (P< 0.0001, FC± South Africa, Mozambique, Kenya, Tanzania 2); 480 upregulated and 588 downregulated and Uganda. A 900bp central fragment of in the C. felis infected samples compared to the p67-encoding gene was amplified from the non-infected controls. Transcripts involved genomic DNA extracted from blood of T. in leukocyte chemotaxis, innate and cellular parva positive samples, cloned and sequenced. inflammatory responses, integrin-mediated Analysis of sequence data revealed four p67 signaling pathway, and phagocytosis were alleles with allele 1 present in both buffalo- upregulated in infected cats, while genes and cattle-derived T. parva strains. Analysis of regulating cell division and double-strand two p67 B cell epitopes in allele 1 sequences break repair via homologous recombination showed that epitope 2 (LQPGKTS) was were downregulated. To the best of our conserved in both cattle- and buffalo-derived knowledge, this is the first report of gene T. parva strains. However, CD isolates had expression in domestic cats in response to unique amino acid substitutions on epitope 1 experimental infection with C. felis. Real time (TEEEVPPADLSDQVL) and within the gene PCR analysis will be conducted to verify region in this allele. These results extend tSMS results. Our preliminary results indicate earlier data suggesting that p67 is a conserved significant shifts in gene expression during C. molecule within cattle-derived parasites felis infection with relevant processes related and reveal differences between p67 allele 1 to inflammation and mitotic activity. variants associated with ECF and CD cases in

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 113 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA30 IAFWP Symposium sample. PCR testing disclosed a 14% (10/72) prevalence of T. gondii in heart muscle. July 9, 2019, 16:30 - 18:00 Our results suggest that T. gondii is highly Breakout Room 5, Meeting Rooms KLOP, Level 4 prevalent in seagulls and waterfowl indicating high environmental contamination in the areas of collection. Results also indicate OA30.01 Unexpected High Toxoplasma that serology is more sensitive than PCR Gondii Prevalence in Waterfowl and Seagulls for detecting T. gondii infection in these bird species. Further research is needed to Dr. Sawsan, Ammar1,6, Janetta Kelly2, Graham elucidate the transmission dynamics of T. Hickling2, Justin Brown3, Kyle Van Why4, Dr. gondii in waterfowl, seagulls and other aquatic Chunlie Su5, Dr. Richard Gerhold1 animals and determine the zoonotic risk. 1Department of Biomedical and Diagnostic Sciences, University Of Tennessee, Knoxville, Knoxville, United States, 2Department of OA30.02 A Duplex PCR for the Simultaneous Forestry, Wildlife, and Fisheries, University Detection of Fasciola Hepatica and of Tennessee, Knoxville, United States, Clonorchis Sinensis 3Pennsylvania Game Commission , Harrisburg, United States, 4USDA APHIS Wildlife Services, Dr. Yi Yang1, Dr. Yimin Yang1, Prof. Aifang Du1 Harrisburg, United States, 5Microbiology 1Institute of Preventive Veterinary Medicine, Department, University of Tennessee, College of Animal Sciences, Zhejiang Knoxville, United States, 6Department of University, Hangzhou, China Clinical Pathology, Faculty of Veterinary Medicine, Sadat City University, Sadat City, Both Fasciola hepatica and Clonorchis Egypt sinensis are endemic in China, South Korea, Japan and other Southeast Asian countries. Toxoplasmosis is a zoonotic disease caused Reliable and sensitive diagnostic methods by the protozoan parasite Toxoplasma gondii are needed for detecting their infections in which infects mammals and birds. Little humans and animals. Differential simplex information is known about the prevalence and duplex polymerase chain reaction and epidemiology of T. gondii in wild birds. (PCR) methods were developed. The PCRs We investigated T. gondii prevalence in targeted the second internal transcribed seagulls and numerous waterfowl species spacer (its2) (408 bp) of F. hepatica, and from Tennessee and Pennsylvania to NADH dehydrogenase subunit 2 gene (nad2) better understand the parasite’s host and (527 bp) of C. sinensis. Both simplex PCRs geographical distribution and zoonotic detected as little as 2 pg genomic DNA in potential. Numerous waterfowl species are one microliter in a 25 L PCR reaction system. game animals and thus represent a potential The duplex PCR had similar detection limit T. gondii zoonotic risk factor whereas as well, and detected μas low as one egg in seagulls are broadly distributed opportunistic 200 mg feces. These methods were analytical feeders in areas of human settlements and specific with no amplification being observed can indicate environmental contamination. from the gemonic DNA of Fasciolopsis buski, We tested a total of 62 serum samples with Haemonchus contortus, Ascaris ovis or Eimeri modified agglutination test and found a ahsata. Of 158 sheep fecal samples collected seroprevalence of 72.6% (45/62). Ringed- from various farms, four and one samples bill gull (Larus delawarensis) samples were PCR-positive for F. hepatica and C. were collected from Pennsylvania and had sinensis, respectively. The duplex PCR method a seroprevalence of 78% (18/23) while described here is time-saving and convenient, waterfowl samples were from hunter-killed and may prove to be an invaluable tool for birds in Tennessee and had a seroprevalence molecular detection and epidemiological of 69% (27/39). In addition, PCR was investigation of F. hepatica and C. sinensis in performed on heart tissues from all of the endemic area. above samples as well as ten heart tissues that did not have a corresponding serum

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 114 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA30.03 Who’s Who in the Poo: A Taqman exposed to both parasites. We also designed Assay for the Differentiation of Fasciola and validated conventional PCR primers for Hepatica, Fasciola Gigantica and Co- either ITS1 or LSU for use on faecal samples infection in Ruminant Faecal Samples to investigate ambiguous results, or animals identified as co-infected. When combined Ms. Nichola Calvani1,2, Dr. Russell Bush2, Dr. with the previously-published workflow, the Jan Slapeta1 result is two assays that provide the first ante- 1Laboratory of Veterinary Parasitology, mortem differentiation between infection with Sydney School of Veterinary Science, F. hepatica, F. gigantica or co-infected animals Faculty of Science, The University of Sydney, in areas of parasite sympatry. Camperdown, Australia, 2Mekong Livestock Research Group, Sydney School of Veterinary Science, Faculty of Science, The University of OA30.04 Study on Zoonotic Parasites in Sydney, Camden, Australia Game in Brandenburg, Germany

Fasciolosis, due to infection with Fasciola Dr. Nadja Bier1, Dr. Claudia Jäckel1, Carolyn hepatica and Fasciola gigantica, is a Kästner1, Prof. Karsten Nöckler1, Dr. Anne neglected zoonotic disease of worldwide Mayer-Scholl1, Dr. Annette Johne1 importance, with economic impacts estimated 1Federal Institute For Risk Assessment, Berlin, to exceed USD $3 billion/year and over 91 Germany million people considered at risk of infection. Infections are commonly diagnosed by a Toxoplasma gondii, Alaria alata, and traditional sedimentation approach, which Cryptosporidium spp. are endemic parasites is time-consuming and prone to sensitivity in Germany. To better estimate the public errors when a large number of samples health risk mitigating from these three must be processed, or if the operator lacks different parasites in wild animals, more data sufficient experience. A commercially- on their presence is needed. available coprological antigen ELISA enables detection of infection prior to the 8-12 week During the hunting season 2017/2018, game pre-patent period, with increased sample was sampled from eight different hunting throughput, however species differentiation grounds in the federal state of Brandenburg, is not possible in areas of parasite sympatry Germany. For the detection of T. gondii in where co-infection with both species may wild boar, roe and red deer, blood samples occur. Diagnosis via real-time PCR offers the from the abdominal cavity were subjected for combined benefits of highly sensitive species serological testing by means of a commercial differentiation for medium to large sample ELISA kit and heart muscle samples were sizes. Despite this, there are currently no examined by qPCR targeting the 529 bp- robust molecular diagnostic assays available repeated element. A. alata was detected in for ante-mortem Fasciola spp. differentiation wild boar using the mesocercariae migration from faecal samples. technique for muscle and fat tissue samples and Cryptosporidium spp. in wild boar, roe Our aim was to address this need via the and red deer was tested in fecal samples by design and validation of two real-time PCR use of 18SrRNA nested PCR. TaqMan assays for Fasciola spp. differentiation using either ITS1 or LSU. We used an existing Serological examination of 194 serum samples diagnostic workflow for faecal sample from roe deer, red deer and wild boar revealed preparation and DNA isolation to achieve T. gondii-specific antibodies in 18.6% of all high analytical sensitivity of <1 egg per gram tested samples. DNA was detected in 26% of faeces. The assays were designed using of seropositive animals but high Cq values samples from 13 F. hepatica-infected sheep may imply a low parasitic burden of T. gondii. and six F. gigantica-infected cattle from Mesocercariae of A. alata could be found in 27% Australia and Laos, respectively, which were of 171 tested wild boar samples and DNA of then used to diagnose infection in 75 cattle Cryptosporidium spp. could be detected by PCR from northern Laos expected to have been in 26.7% of 210 wild boar and deer samples.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 115 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

These results show a high exposure of game pyrosequencing, and applying the method to to T. gondii, A. alata and Cryptosporidium populations containing known proportions spp. in the monitored hunting areas in of resistant and susceptible L3. Finally, we Brandenburg, Germany and may indicate applied the method to field samples collected possible transmission routes of these parasites from ewes and lambs on over a period of to humans. one year on three farms, each highlighting different aspects of sheep management and approaches to parasite control. The results OA31 Anthelmintic Resistance in show opportunities to build hypotheses with Ruminants reference to selection pressures leading to differences in resistance allele frequencies July 10, 2019, 11:00 - 12:30 between sampling dates, farms and ewes or Plenary Hall, Madison Ballroom (ABCD), Level 4 lambs, and to consider the impact of their genetic fixation or otherwise. This study provides proof of concept of a practical, OA31.01 Development of High Throughput accurate, sensitive and scalable method to Method for the Analysis of Anthelmintic determine frequency of anthelmintic drug Resistance Allele Frequencies in Field resistance mutations in gastrointestinal Populations of Gastrointestinal Nematodes nematodes in field studies and as a management tool for livestock farmers. Umer Chaudhry1, Madison MacLeay1, Alison A. Morrison2, David J. Bartley2, Mike Evans1, Neil D. Sargison1 OA31.02 A 16-Year Retrospective Analysis 1University Of Edinburgh, United Kingdom, of Anthelmintic Resistance in Haemonchus 2Moredun Research Institute, United Kingdom Contortus on Small Ruminant Farms in the United States Drug resistant helminths have become a major cause of poor health and production Mrs. Sue Howell1, Mr. Brandon Park2, Dr. Anand in ruminanats, and there is a need for Vidyashankar2, Mr. Bob Storey1, Mr. James diagnostic markers and tools to determine the Collins1, Dr. Ray Kaplan1 frequency of resistance alleles in field parasite 1University Of Georgia-Dept. of Infectious populations. Gastrointestinal nematode Diseases, Athens, United States, 2George resistance to benzimidazole drugs is caused Mason University-Dept of Statistics, Fairfax, by a mutation in one of three positions on the United States isotype 1 -tubulin locus, and in the absence of markers for resistance to other broad- The DrenchRite© larval development assay β spectrum anthelmintic classes, these provide (LDA) measures resistance to benzimidazoles, a relevant study example. Determination of levamisole, ivermectin, and moxidectin. From the prevalence of these single nucleotide 2000-2016 we performed DrenchRite© polymorphisms in field gastrointestinal LDA on 291 small ruminant farms in 40 US nematode populations can be impractical states. We then performed a retrospective using conventional molecular methods, which analysis of these data to investigate changes may be error prone or lack sensitivity at in prevalence and levels of anthelmintic low levels of resistance. Here, we report the resistance in Haemonchus contortus over development of a novel method based on an this period. For analyses, time series models Illumina Mi-seq deep amplicon sequencing were used to identify shifting and emerging platform; to sequence the isotype 1 -tubulin resistance trends across time, and data were locus of the small ruminant gastrointestinal grouped into three 5-6 year intervals (2000- β nematode, Teladorsagia circumcincta, and 2005; 2006-2010; 2011-2016). Resistance determine the frequency of the benzimidazole was highly prevalent to benzimidazoles resistance mutations. We validated the and ivermectin in goats reaching 100% method by assessing sequence representation and 95% prevalence, respectively by 2011- bias in the isotype 1 -tubulin locus, 2016. Prevalence of levamisole resistance comparing the results of Illumina Mi-seq and was consistently the lowest of all drugs on β

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 116 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

both goat and sheep farms. The greatest extent of AR in the Irish cattle population. A changes were for moxidectin, and these were total of 24 dairy-to-beef farms were recruited significantly greater for goat than sheep for the study and on each farm groups of 20 farms (p<0.0001). Prevalence of resistance to first grazing season calves were treaded with moxidectin progressively increased over the either benzimidazole (BZ), levamisole (LEV), 3 time intervals for goat (28%,38%,56%) and ivermectin (IVM) or moxidectin (MOX) (not sheep (3%,18%,40%) farms, with mean IC50 all drugs were tested on every farm). Faecal (nM) increasing from 592,225 (2000-2006) egg count reduction was determined from to 2433,1761 (2011-2016) for goat and sheep pre- and post- treatment faecal samples. farms, respectively. The time series model Faecal egg counts were determined using the for moxidectin on goat farms confirmed mini-FLOTAC technique (sensitivity of 5 eggs that low resistance was initially detected, per gram). Pooled pre- and post-treatment followed by full resistance during the later faecal cultures were also established. Reduced years. For sheep farms, an increasing trend BZ efficacy (range 15%-93%) was recorded was demonstrated where susceptibility on 71% (12/17) of farms; 25% (3/12) of the was shown initially, which then moved to farms recorded reduced efficacy for LEV suspected resistant in the middle-years, and is (range 79%-90%). All the farms tested (100%; currently moving toward full-resistance. Total 17/17) showed reduced IVM efficacy (range anthelmintic failure (resistance to all 3 classes -228%-89%) and 75% (9/12) of the farms plus moxidectin) increased over time from recorded reduced efficacy against MOX 3% to 35% and 3% to 10% for goat and sheep (range 1%-93%). Cooperia and Ostertagia farms, respectively. These data and the time were the predominant nematode genera series analyses performed provide interesting present on all farms. Surprisingly, Ostertagia insights into changes occurring in resistance was identified as the main nematode species levels over time, and differences in changes (> 50%) surviving treatment in 89% BZ, 33% between drugs and hosts. Though worse on LEV, 23% IVM and 12.5% MOX resistant farms. goat than on sheep farms, these data indicate Results from this study indicates that AR is a severe situation exists in the USA with widespread on cattle rearing farms in Ireland regard to anthelmintic resistance. and that strategies to manage and minimise AR are urgently needed.

OA31.03 Evidence of Widespread Anthelmintic Resistance in Cattle in Ireland OA31.04 Nicotine-Sensitive Acetylcholine Receptors Are Relevant Pharmacological Ms. Anne Kelleher1,2, Dr. Orla Keane2, Dr. Targets for the Control of Multidrug Resistant Barbara Good3, Assoc. Prof. Theo de Waal1 Parasitic Nematodes 1University College Dublin, Belfield, Ireland, 2Teagasc, Animal and Bioscience Department, Dr. Elise Courtot1, Fabrice Guégnard1, Jacques Grange, Ireland, 3Teagasc, Animal and Cortet1, Dr. Cédric Neveu1, Dr. Claude Charvet1 Grassland Research and Innovation Centre, 1INRA, Université de Tours, UMR1282, 37380, Athenry, Ireland Nouzilly, France

Irish beef production is predominantly The control of parasitic nematodes impacting pasture based with the result that grazing animal health relies on the use of broad cattle are exposed to a number of different spectrum anthelmintics. However, intensive gastrointestinal nematodes. Irish farmers rely use of these drugs has led to the selection heavily on anthelmintic treatments to control of resistant parasites in livestock industry. helminth infection. Although anthelmintic In that respect, there is currently an urgent resistance (AR) is more commonly reported need for novel compounds able to control in sheep it has been increasingly reported in resistant parasites. Nicotine has also cattle worldwide. In 2014 AR was reported for historically been used as a de-wormer until the first time on two cattle farms in Ireland. modern anthelmintics were marketed. The During 2017 and 2018 a more extensive survey pharmacological target of nicotine has been was conducted in Ireland to determine the identified in nematodes as acetylcholine-

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 117 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

gated ion channels. Nicotinic-sensitive with different mechanisms of action has acetylcholine receptors (N-AChRs) therefore been proposed. A pharmaco-parasitological represent validated pharmacological targets assessment of different nematodicidal than remain largely under-exploited. combinations was performed at different In the present study, we developed an cattle commercial farms in Argentina. The automated larval migration assay (ALMA) Ivermectin(IVM)-ricobendazole(RBZ), and showed that nicotinic derivatives IVM-levamisole(LEV) and RBZ-LEV (anabasine/nornicotine) efficiently paralyzed combinations were assessed in calves a multiple (benzimidazoles/levamisole/ naturally infected with susceptible/resistant pyrantel/ivermectin) resistant field isolate gastrointestinal nematodes. The observed of Haemonchus contortus. Additionally, pharmacokinetic(PK) data demonstrated that using Caenorhabditis elegans as a model, the co-administration of two anthelmintics we confirmed that the N-AChRs subtype did not modify the plasma PK behaviour contributes to the anthelmintic effect of of either drug in cattle. In fact, no adverse nicotinic analogs. Interestingly, the functional PK interactions were observed after expression of the homomeric N-AChR from each combined treatment. Similar PK C. elegans and the distantly related horse parameters(P>0.05) were obtained between parasite Parascaris equorum in Xenopus the single-drug and the combined-based oocytes highlighted some striking differences strategies. The IVM-RBZ, IVM-LEV and RBZ- in their respective pharmacological properties LEV combinations were the only treatments towards nicotine derivative sensitivity. achieving 100% clinical efficacy, even when a Noteworthy, nicotine and anabasine were highly IVM-resistant Haemonchus spp. isolate more potent than ACh in activating the was present(45% efficacy for IVM alone). In P. equorum N-AChR as revealed by their fact, the combination IVM-RBZ maintained respective EC50 values (2.9 ± 0.5 M and its 100% efficacy against an IVM-resistant 1.7 ± 0.1 M versus 6.4 ± 1.1 M, respectively), Haemonchus spp. population over the last unlike nornicotine (34.9 ± 7.2 M) whereasμ the five years. Additionally, the weight gain was potencyμ series for the C. elegansμ N-AChR was significantly higher in calves treated with Nic > ACh = Ana > Nor. μ the IVM-RBZ combination. Overall, after the Taken together these results validate the combined treatments only a therapeutic exploitation of the N-AChRs of parasitic additive effect is achieved. However, the nematodes as targets for the development of efficacy of IVM-RBZ against nematodes resistance-breaking compounds. resistant to IVM and RBZ was greater than an additive effect. In fact, in two farms with multiple resistance to IVM and RBZ, while the OA31.05 Anthelmintic Combinations: A efficacies during the first year of study were Sustainable Strategy to Optimize Parasite 54%(IVM), 84%(RBZ) and 98%(IVM-RBZ) in Control on Commercial Cattle Farms? Farm A, the egg reductions were 40%(IVM), 64%(RBZ) and 90%(IVM-RBZ) in Farm B. Dr. Candela Canton1, Dr. Laura Ceballos1, During the second year of study, the high Dr. Laura Moreno1, Veterinarian María Paula efficacy of the combination IVM-RBZ could be Domínguez1, Ms. Lucila Canton1, Veterinarian maintained in Farm A(97%), but not in Farm Miguel Buffarini2, Dr. Carlos Lanusse1, Dr. Luis B(67%). Overall, the potential advantages Ignacio Alvarez1 of anthelmintic combinations should be 1Laboratorio de Farmacología, Centro de carefully assessed before being extensively Investigación Veterinaria de Tandil (CIVETAN), recommended. Their rational use should UNCPBA-CONICET-CICPBA, Tandil, Argentina, be strongly supported by pre-treatment 2EEA INTA Gral. Villegas, Gral. Villegas, diagnosis and considering the epidemiological Argentina situation of each individual farm.

In an attempt to minimize therapeutic failures following anthelmintic treatments and to delay the development of resistance, the combined use of nematodicidal compounds

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 118 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA32 Avian Parasites the water (SafeGuard® Aquasol, 1mg/kg) for five consecutive days. Water consumption, July 10, 2019, 11:00 - 12:30 feed intake, and bird weights were measured Breakout Room 2, Hall of Ideas E&H, Level 4 weekly. Differences between treated and untreated groups of an isolate, as well as between the two isolates, are being analyzed OA32.01 Resistance to Fenbendazole in to determine the impact of resistant parasites Ascaridia Dissimilis and Its Economic Impact feed consumption, water consumption and of the Production of Turkeys feed conversion.

Mr. James Collins1, Dr. Brian Jordan1, Dr. Luke Baldwin1, Dr. Justin Fowler1, Dr. OA32.02 Methodology for the Claude Hebron2, Dr. Kelsey Paras1, Dr. Anand Assessment of Parasitic Infections Vidyashankar3, Dr Ray Kaplan1 Caused by Allopsoroptoides galli (Acari, 1University of Georgia, Athens, United States, Psoroptoididae) in Laying Hens 2Prestage Farms, Clinton, United States, 3George Mason University, Fairfax, United Edna Clara Tucci1, Nilce Maria Soares2, Paulo States Henrique Selbmann Sampaio1 1Animal Health Research Center, Biological Ascaridia dissimilis is the most prevalent Institute, São Paulo, Brazil, 2Specialized and one of the most economically important Poultry Health Laboratory, Biological Institute, gastrointestinal nematodes of turkeys. Bastos, Brazil Infections are most often subclinical, producing reduced feed conversion Allopsoroptoides galli is a parasitic mite efficiency, with heavier infections causing found in 2010 when it shifted from wild avian clinical symptoms such as lethargy, intestinal hosts to battery caged layers in the state of blockage, diarrhea, and in some cases São Paulo, Brazil. Clinical findings include death. Currently, fenbendazole is the only intense pruritus and severe dermatitis, with drug approved by the United States Food dry scaling skin and yellowish exsudate on and Drug Administration for use against the calami. Necessity prompted standardizing this parasite in turkeys. We recently tested a methodology to study the infection. It is the efficacy of fenbendazole against 4 based on grading dermatitis of the apteria field isolates of A. dissimilis acquired from and counting live mites on skin scrapings. To commercial farms. Three of the four isolates grade dermatitis, a mean score of the ventral (Wi, Ow, Po) demonstrated greater than 99% alar, dorsal and lateral apteria is calculated. efficacy, with the fourth (Sn) yielding only Scores range from 0 to 5 (0 = no dermatitis; 63.89% efficacy, indicating that this isolate 1 = mild dermatitis; 2 = mild to moderate is resistant to fenbendazole. Having proven dermatitis; 3 = moderate dermatitis; 4 = fenbendazole resistance in A. dissimilis, we moderate to severe dermatitis and 5 = severe wanted to determine the economic impact dermatitis). Microscope slides are used to that resistant worms have on growth and scrap each apterium until a hemorrhage productivity. We infected 384 turkey poults occurs. Scrapings are clarified with potassium with either the Sn resistant isolate or the hydroxide, coverslips are placed and acari are Ow susceptible isolate and divided them immediately counted under a microscope. into treated and untreated groups. Both the To validate the methodology, 22 naturally treated and untreated groups for each isolate infected layers were randomly assigned to had 8 replicates of 12 birds each. Turkeys were treatment with a 3% emulsion of a botanical grown for an 18-week growing cycle, changing acaricide (70% Azadirachta indica; 2.5% feed types at the recommended time points Melaleuca alternifolia and 2.5% Melaleuca as per normal commercial growing practices. leucadendron essential oils) or to the control Birds were infected via a trickle dosage of group. 0.3% neutral detergent was added to 50 eggs/bird/week sprinkled onto their feed. aid skin contact. Control fowl were sprayed At 4, 8, 12 and 16 weeks, birds in the treated with water and 0.3% neutral detergent. Both groups were administered fenbendazole in groups were sprayed on experimental days 0,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 119 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

+7 and +14. Hens were checked for dermatitis component. On-animal sensors, which measure and mite counts on days +7; +14; +28; +35; magnitude of force (‘movement’) along 3-axes, +42 and +72. A significant reduction of acari were used to identify and quantify chicken was achieved after the first treatment, but behaviors before and during ectoparasite mites were recovered until day +35. Treated infestation compared to uninfested control layers presented mild dermatitis until the birds. The behaviors measured were pecking, end of the trial. Dermatitis does not imply preening, and dustbathing. Allopsoroptoides infection. Therefore, checking apteric skin for mites is fundamental. During a pre-infestation period (week 1), The standardized methodology presented in both WQA measures and bird behavior data the current abstract is effective and can be (sensor readings) was recorded to provide useful in further studies, under experimental baseline data for bird welfare and behavior. and field conditions. Lice were subsequently introduced to 2 of 4 flocks, and WQA and sensor readings were repeated during Week 7 and Week 12 after OA32.03 On-Animal Sensors to Assess louse populations had established in the Chicken Body Louse Effects on Poultry infested flocks. Behavior and Welfare Variation of WQA and behaviors among Dr. Amy Murillo1, Dr. Richard Blatchford2, Mr. individuals is high, though preliminary Alireza Abdoli1, Dr. Eammon Keogh1, Dr. Alec results indicate trends that relate to louse Gerry1 populations on birds. 1University of California Riverside, Riverside, United States, 2University of California Davis, Davis, United States OA32.04 More Effective Control of House Flies in Poultry Facilities Using the Fungus The chicken body louse (CBL), Menacanthus Beauveria Bassiana stramineus, is a common poultry pest in the United States, especially on small backyard Alexandra Pagac1, Dr. Erika Machtinger1, Dr. or hobby flocks. It is an obligate ectoparasite Christopher Geden2 that primarily feeds on feathers, but will also 1Penn State University, University Park, United blood-feed. Severe infestations can cause States, 2USDA, Gainesville, United States economic damage to egg-laying hens. Control of house flies and other muscids are Increasing awareness of animal welfare in a significant challenge for poultry producers, livestock has led to the development of especially in layer facilities. Most egg standardized metrics for assessing animal production occurs indoors with accumulating welfare. One example, the Welfare Quality® poultry waste thus creating perfect habitat for Assessment (WQA) protocol has been widely breeding flies. To control pest flies, the poultry adopted in North America as a research and industry spends 20 million dollars annually on-farm tool. The WQA uses both environment- on pesticides alone. This estimate doesn’t and animal-based measures, but only the include the cost of animal loss due to house presence or absence of ectoparasites is noted fly vectored pathogens causing disease, the and there is no qualitative or quantitative cost of labor for pesticide application, or evaluation. The degree of impact ectoparasites litigation that can be taken by residents living have on hen health or welfare is likely to vary near production facilities due to increased fly based on ectoparasite species and severity of numbers. However, regulatory restrictions, infection. In this study, animal welfare metrics fly resistance to current active ingredients, were evaluated as a function of louse infestation and lack of options labeled for pest control in using two groups of birds; louse infested flocks poultry facilities make fly control challenging. and louse-free control flocks. Finding a new and effective tool for fly control is needed by the poultry industry. Biological Louse infestations may also hinder or alter control using the entomopathogenic fungi normal chicken behaviors, another welfare Beauveria bassiana may be an effective

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 120 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

method for fly control. One product is oocyst scores, bloody diarrhea and FCR in currently available for use in poultry facilities, chicks treated with herbal extracts compared but efficacy has been unsatisfactory. In with infected unmedicated chicks. The addition, B.bassiana takes 6 or more days survival rate and weight gain was higher in to kill an adult fly which limits the ability chicks treated with herbal extract compared to control house fly outbreaks. The goal of with infected unmedicated chicks. According this project is to develop a more effective B. to the findings of this study, the HF is suitable bassiana product and develop it into a more for the resource-poor farmers as a broad practical tool that can be easily adopted spectrum antiparasitic. The contents of the into both Integrated Pest Management formulation are cheap, commonly available, programs and organic farming practices. and easy to use as a decoction. Incorporation The objectives are to 1) identify isolates of of this herbal formulation in integrated B. bassiana from house flies collected from parasite management practices will add to poultry facilities in Pennsylvania, 2) select the sustainability and thus, income of the successful isolates for quick kill times, and farmers. The HF seems promising as a broad 3) assess the compatibility of these isolates spectrum antiparasitic. Large scale controlled on other common biological control agents studies are, however, recommended for (e.g., parasitoid wasps). House fly collections standardization of the doses and applications were made on 8-layer house facilities in of the product. Future experiments may Pennsylvania bi-weekly from June to August. be planned to understand the mechanism To date, five isolates of B. bassiana have been of absorption as well as developing an identified from fly collections. Selection and appropriate delivery agent for the herbal safety results will be discussed. formulation inside the tick body. Studies on shelf life of the plant extract and its residual activity need to be carried out. OA32.05 Effect of a Herbal Complex Against Common Parasites of Animals and Poultry in Pakistan OA32.06 Georeferenced Database of Avian Wild Parasites in Mexico Dr. Muhammad Arfan Zaman1, Professor Zafar Iqbal2, Dr. Rao Zahid Abbas2, Professor Cecilia Marina Vázquez Vega1 Muhammad Fiaz Qamar1 1Department of Parasitology of the Faculty 1College of Veterinary and Animal Sceinces, of Veterinary Medicine (FMVZ), Ciudad Jhang, Pakistan, 2Department of Parasitology, Universitaria, Mexico University of Agriculture, Faisalabad, Pakistan Georeferenced anaylisis techniques emerged, The objective of the current study was to among other causes, to relate environmental, evaluate the anti-tick, anthelmintic and biological and abiotic variables, with the anticoccidial activities of a herbal formulation epidemiology of diverse infectious diseases. (HF) based on complex of water extracts of There is a great diversity of documented avian leaves of Azadirachta indica and Nicotiana parasites in Mexico. However, the information tabacum, flowers of Calotropis procera is dispersed in different documents, making and seeds of Trachyspermum ammi. The the research even more variable. HF demonstrated anti-tick activity by inhibiting the egg laying, larval mortality The main objective of this work consisted of and reduced tick intensity/infestation on collecting, purifying, organizing and analyzing animals. Anthelmintic activity of herbal the information about wild avian parasite formulation was evident from the in vitro occurrences and presences from bibliographic mortality of Haemonchus contortus, ovicidal and diagnostic data all over the country for effects in egg hatch test and fecal egg count the development of a georeferenced database reduction in sheep naturally parasitized with illustrated with several maps to show the gastrointestinal nematodes. Anticoccidial parasite occurrences of these registered effects of herbal formulation were confirmed hosts in Mexico from 1990 to 2018, as well as by reduction in the oocyst counts in feces, the projected distribution to the year 2050

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 121 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

considering the effects of climate change. diagnosis of Anoplocephalidae and Fasciola A total of 982 records were integrated hepatica, respectively. Faecal cultures were into the data base and 7 maps of parasites performed for each horse with strongyles occurrences were generated corresponding to faecal egg count (FEC) >200 EPG. Intestinal 279 municipalities, as well as 23 maps of the strongyles were the most common parasitic projected distribution to 2050 according to species found with a prevalence of 54.5%. the Maxent program. The bioclimatic variables Infection intensity was lower than 200 EPG that contributed to the parasite distribution in 27.5% of horses (low contaminators); are BIO19, BIO15, BIO2 and BIO4. 11.3% were moderate contaminators (200- The knowledge of the environmental variables 500 EPG), 15.7% were high contaminators involved in the spatial distribution of the (>500 EPG); 45.5% had negative strongyle parasites will support the proposal of parasite egg count. Faecal cultures revealed only the infection risk in avian wildlife to apply One presence of cyathostomins. Other parasites Health guidelines an approach to design were Parascaris spp. (9.7%), Anoplocephala and implement preventive and conservation spp. (4.1%), Oxyuris equi (3.2%), Eimeria strategies and programs. leuckarti (0.6%) and Strongyloides westeri (0.1%). No eggs of Fasciola hepatica were detected. Prevalence of strongylid infection OA33 Equine Cyathostomes I was significantly different for horse ≤1 year (OR=2.02, p<0.0001) and 1-4 years July 10, 2019, 11:00 AM - 12:30 PM (OR=1.99, p<0.0001) versus older horse Breakout Room 3, Hall of Ideas F&I, Level 4 (>4 years). Prevalence of ascarid infection was significantly different for horse ≤1 year (OR=56.95, p<0.0001) and 1-4 years OA33.01 A Coprological Survey on Internal (OR=11.62, p<0.0001) versus older horse. Parasites of Horses in Italy Considering the high prevalence of horses

1 1 with FEC<200 EPG (73%) the responsible Dr. Francesco Buono , Dr. Laura Pacifico , use of equine anthelmintics is mandatory. Dr. Diego Piantedosi1, Giovanni Sgroi1, Dr. 2 3 In practice it is necessary to determine the Domenico Rufrano , Dr. Cristina Roncoroni , EPG using a quali-quantitative coprological Dr. Giuseppe Mazzeo4, Dr. Francesco 4 1 method and treatments should be made only Chiarotto , Dr. Vincenzo Veneziano after a diagnosis. It is crucial that veterinary 1University of Naples Federico II, Napoli, play an active role in planning and monitoring Italy, 2Research Unit for the Extensive Animal effective and appropriate equine worm Husbandry, Muro Lucano (PZ), Italy, 3Istituto control programs. Zooprofilattico Sperimentale Lazio e Toscana, Rome, Italy, 4MSD Animal Health S.r.l., Segrate (Milan), Italy OA33.02 Precision, Sensitivity, and Specificity Analysis of an Automated Parasite Internal parasites are ubiquitous in horses and Fecal Egg Counting System in Comparison to cyathostomins are the most important due to McMaster and Wisconsin Methods the development of anthelmintic resistance. A national survey was conducted from Jennifer Cain1, Paul Slusarewicz2, Morgan December 2013 to January 2019 to assess McVey1, Kayla Wielgus3, Haley Zynda1, Libby the prevalence of the main endoparasites Wheling1, Eric Roemmele1, Dan Lin4, Martin in 338 horse farms in Italy. Faecal samples Nielsen1 were collected from 3,092 horses (764 1University Of Kentucky, Lexington, United males, 1592 females, 736 geldings), mean States, 2MEP Equine Solutions, Lexington, age 9.9 (1 month–34 years) and examined United States, 3Lincoln Memorial University, using the Mini-FLOTAC technique (detection Harrogate, United States, 4Hasselt University, limit: 5 eggs per gram) and a Sheather’s Antwerp, Belgium sugar solution (specific gravity:1.250). A centrifugation/flotation technique and a Fecal egg counts are the cornerstone of sedimentation technique were used for the equine parasite control programs. Previous

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 122 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

work developed an automated image analysis Strongylus vulgaris is widely recognized as based parasite egg counting system that a major pathogen in horses, causing non- reduces operator error, and should thus strangulating intestinal infarctions, peritonitis, increase method precision. The system and death. In addition, cyathostomin has been further developed to include an (small strongyle) parasites are ubiquitous automated reagent dispenser unit and a in grazing horses and are main targets of standalone digital imaging unit generating parasite control in mature horses. Historic higher resolution images. The aims of this studies conducted in Northern Europe have study were to conduct a comprehensive suggested seasonal fluctuation in strongyle comparison of method precision between egg shedding as well as the occurrence of two different imaging units as well as the patent S. vulgaris infection; however, these traditional McMaster and Wisconsin manual epidemiological patterns have not been techniques, and to perform a Bayesian analysis investigated in recent years or in other for estimation of method sensitivity and climatic locales. In this study, we analyzed specificity. Feces were collected from horses, the seasonal fluctuation of strongyle fecal screened with triplicate Mini-FLOTAC counts, egg counts, presence of S. vulgaris larvae on and placed in very high (>1000 eggs per gram coproculture, and concentration of antibodies (EPG)), high (500-1000 EPG), and medium specific to migrating S. vulgaris larvae in a (>500-200 EPG) egg count categories. Ten research herd of horses maintained at the replicates per horse were analyzed for each University of Kentucky that has not received technique: the automated system using anthelmintic treatment since 1979. Fecal both a smartphone and standalone camera and serum samples were collected from all model, McMaster, and Wisconsin. Although horses (N=20) twice monthly during 2018. not statistically significant from McMaster Individual strongyle fecal egg counts and (76.1%), the standalone camera produced coprocultures were carried out for every the numerically highest (78.8%) precision for time point. Similarly, a validated ELISA was the high counts. Wisconsin had the lowest run to determine the concentration of anti-S. precision at 63.7%, and the smartphone vulgaris antibodies. Data were analyzed with camera had a precision of 68.7%. Wisconsin mixed linear models. There were no seasonal tended to produce lower egg counts than the differences in strongyle egg shedding and other methods, whereas the older camera anti-S. vulgaris antibody levels; however, system tended to produce higher counts than the number of S. vulgaris third stage larvae the others. The Bayesian analysis also included identified in coprocultures was significantly low (> 200 - < 0 EPG) and negative (no eggs higher (p>0.0001) in the spring compared seen) counts, and results for sensitivity and to summer and autumn. Furthermore, horses specificity will be presented at the conference. aged 15-18 years of age had significantly Overall, the automated counting method higher S. vulgaris larval counts than horses is a promising new development in equine aged 8-10 years (p<0009). The lack of parasitology, and continuous improvement seasonality in strongyle egg shedding is to the counting algorithms will help increase in contrast with studies performed in the precision in the future. United Kingdom several decades ago. Possible reasons could be climatic influences, cyathostomin species composition, stocking OA33.03 A Parasite for all Seasons: density, and the absence of anthelmintic Investigation of Seasonality in Strongyle Egg treatment. It was surprising to see the oldest Shedding and Strongylus Vulgaris horses demonstrate the highest S. vulgaris larval counts, as this parasite is generally Haley Anderson1, Dr. Ashley Steuer1, Jessica considered to be immunogenic. Kenealy1, Taylor Shepherd2, Morgan Clark2, Holli Gravatte1, Dr. Martin Nielsen1 1University of Kentucky, Maxwell H. Gluck Equine Research Center, Lexington, United States, 2College of Veterinary Medicine, Lincoln Memorial University, Harrogate, United States

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 123 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA33.04 It’s All About the Mucus? Evaluation OA33.05 Stronylus Vulgaris and Colic in of the Immune Response to Larvicidal Swedish Horses: A Case-Control Study Treatment of Equine Cyathostomin Infection Ass Prof, Eva Tydén1, Dr. Ylva Hedberg- Dr. Ashley Steuer1, John Stewart1, Day Barker1, Alm1, Dr. Miia Rhimäkii1, Dr. Martin Nielsen2, Dr. Amanda Adams1, Dr. Martin K. Nielsen1 Professor Arvid Uggla1, Eva Osterman-Lind3 1University Of Kentucky, Maxwell H. Gluck 1Swedish University of Agricultural Sciences, Equine Research Center, Lexington, United Uppsala, Sweden, 2Associate professor, M.H. States Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Cyathostomins are omnipresent parasites Lexington, United States, 3National Veterinary of equids worldwide. In rare cases, they can Institute, Department of Microbiology, cause life-threatening larval cyathostominosis, Uppsala, Sweden which is characterized by a generalized typhlocolitis when synchronous mass Strongylus vulgaris is re-emerging in the emergence of the larvae from the large Scandinavian horse population with a intestine occurs. Little is known about the prevalence of about 60% on farm level. In immune response to these parasites in horses. recent years an increase in parasite-related Goblet cell hyperplasia has previously been injuries, such as higher frequency of colics noted as an important component with and parasite related damages at autopsy, cyathostomin infection; however, its role is have been noted at the clinics of the Swedish unclear. This study was to evaluate the local University of Agricultural Sciences (SLU). The and systemic inflammatory response, with aim of this study was to evaluate if S. vulgaris particular interest in the role of goblet cells, is more prevalent in horses with clinical signs to cyathostomin infections following larvicidal of colic. This was investigated in a case- treatment. 36 ponies with naturally acquired control study conducted from February 2017 cyathostomin infections were evenly divided to February 2018 at the equine clinic of SLU. into three groups: fenbendazole (10mg/ Cases were horses with signs of colic and kg PO 5days), moxidectin (0.4mg/kg PO controls were horses that attended the clinic once), and untreated control. Blood was on the same week but for other reasons. collected weekly and tissue collected from Blood and faecal samples were collected the cecum and dorsal and ventral colon at from 142 cases and 142 controls. Sera were 2 and 5 weeks post treatment (WPT). Real- analysed by a S. vulgaris antigen ELISA and time PCR was used to evaluated the gene faecal samples for strongyle eggs with a expression of IL-4, IL-5, IL-6, IL-10, IL-13, IL-22, modified McMaster method and for S. vulgaris IFN , Resistin-like Molecule beta (RELM ), with an ITS-2 specific PCR on larval cultures. Mucin 2 (MUC2), and TNF . MUC2, RELM , No significant associations were found andγ IL-22 were decreased in circulation β between colic and positive serology, level of at 3-5 WPT (p<0.0001), suggestingα a roleβ faecal egg output or S. vulgaris positive larval and spillover involving goblet cells across cultures. Eight horses with colic underwent all groups. Pro- and anti-inflammatory surgery of which one had suspected signs of markers were significantly different between migrating S. vulgaris larvae due to inflamed treatment groups in tissues. These results mesenteric arteries. To be highlighted, four suggest that larvicidal treatment can affect horses with colic were euthanized and after the mucosal immune response itself, as necropsy two of these were confirmed to have well as immune modulation elicited by intestinal infarction caused by migrating S. cyathostomin larvae. We also found evidence vulgaris larvae in the mesenteric arteries. Both that the large intestinal organs respond these horses excreted 0 strongyle eggs and differently to treatment and to the encysted were negative on the ITS-2 PCR but positive larvae themselves. These results may have in the ELISA. To conclude, from this study we implications for unraveling parasitic disease could not demonstrate a higher occurrence of processes and consequences of larvicidal S. vulgaris in horses with clinical signs of colic treatment. than in control horses without colic.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 124 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA33.06 Development of a Diagnostic Test specificity). Following optimisation of for Small Strongyle Infections in Equines – recombinant antigen expression and Supporting Treatment Decisions in Practice purification, we are now validating a commercial version of the test. Dr. Kathy Geyer1, Dr. Kirsty Lightbody2, Dr. Tom Tzelos3, Prof Martin Nielsen4, Dr. Corrine The availability of such a test for detection Austin2, Prof Jacqui Matthews1 of cyathostomin infection will provide an 1Moredun Research Institute, Edinburgh, important tool to inform veterinarians on United Kingdom, 2Austin Davis Biologics, treatment decisions and contribute toward Great Addington, United Kingdom, 3The sustainable control of equine small strongyle Roslin Institute, Edinburgh, United Kingdom, infections. 4GLUCK Equine Research Centre, University of Kentucky, United States OA34 Flies and Fly Control in Small strongyles (cyathostomins), comprising Ruminants a group of approximately 50 different species, are universally regarded as the most July 10, 2019, 11:00 - 12:30 prevalent and potentially pathogenic helminth Breakout Room 4, Hall of Ideas G&J, Level 4 parasite of equines. Cyathostomin early third stage larvae (L3) can undergo a period of developmental arrest by encystment in the OA34.01 Impacts of Long-Term Insecticide mucosa/submucosa of the gut wall where they Treatment Regimes on SKRD and KDR can remain inhibited for months to years. The Pyrethroid Resistance Alleles in Horn Fly clinical syndrome, larval cyathostominosis, Field Populations is caused by en masse development and Dr. Luísa N Domingues1, Dr. Felix D Guerrero1, emergence of encysted larvae from the gut 2 wall and has a case fatality rate as high as 50%. Dr. Lane D Foil 1USDA-ARS Knipling-Bushland U. S. Livestock A diagnostic test that informs on cyathostomin Insects Research Lab, Veterinary Pest burden, especially levels of encysted larvae, is Genomics Center, Kerrville, United States, lacking. In some regions, an annual moxidectin 2Department of Entomology, Louisiana State (MOX) treatment in autumn/winter is the University, Baton Rouge, United States current recommended approach to target cyathostomin larvae in all horses. Due to extensive use of anthelmintics in interval- We evaluated the effects of four different treatment programmes, drug resistance is six-year duration control strategies on a major problem, as evidenced by reported the resistance levels and frequency of the widespread resistance of cyathostomins to pyrethroid target site resistance alleles, fenbendazole and pyrantel compounds. As superkdr (skdr) and kdr, at four field reduced cyathostomin egg reappearance populations of Haematobia irritans irritans period (ERP) after MOX treatment is now (Linnaeus, 1758) (Diptera: Muscidae) in commonly reported, it is paramount to Louisiana, USA. Consecutive use of pyrethroid preserve efficacy of this anthelmintic, the only ear tags for six years caused a significant remaining effective larvacidal compound. increase in the resistance ratio to pyrethroids With the aim of developing a serum-based as well as the frequencies of both skdr and ELISA for diagnosis of cyathostomin infection, kdr resistance alleles. After three years of we previously identified diagnostic antigens consecutive use of pyrethroid ear tags, from the common cyathostomin species, followed by one year with no treatment, and Cylicostephanus longibursatus, Cyathostomum followed by two years with organophosphate catinatum and Cylicocyclus nassatus. Following ear tags, the resistance ratio for pyrethroid the production of recombinant proteins for use was not significantly affected, the %R-skdr in ELISA and immunoblotting, these antigens significantly dropped while the %R-kdr allele have been shown to be targets of a specific remained relatively high and stable. Similar IgG(T) response. The ELISA demonstrates results were observed when pyrethroid ear great diagnostic potential (93% sensitivity/88% tags were used for three consecutive years,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 125 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

followed by one year with no treatment, Mato Grosso do Sul state, Brazil) and three and followed by two years with endosulfan wild populations collected at sugarcane mills ear tags; however, this treatment resulted in in the state of Mato Grosso do Sul. Wild flies a slight increase in the resistance ratio for were collected at sugarcane mills with Nzi pyrethroids. In a mosaic, the resistance ratio traps and immediately used in bioassays. for pyrethroids showed a 2.5-fold increase but Susceptibility to cypermethrin was assessed the skdr-kdr genetic profiles did not change, after a 2-hour exposure to impregnated filter as the %R alleles (skdr and kdr) remained papers. All tested populations were resistant to low and stable through the six years. Lack cypermethrin, with resistance factors ranging of exposure to pyrethroid insecticides from 6.8 to 38.6 in the wild populations, for three years significantly affected the showing that pyrethroid resistance is currently skdr mutation but not the kdr mutation, present in stable fly populations in the Brazilian preventing re-establishment of susceptibility Midwest. To our knowledge, this is the first to pyrethroids. SS-SR (skdr-kdr) individuals report of insecticide resistance in stable flies in were responsible for the maintenance of Latin America. the kdr mutation in two of the populations studied, and fitness cost seems to strongly affect the SR-RR genotype. None of the four OA34.03 Selected Insecticide Delivery treatment regimens evaluated in the study Devices for Management of Horn Flies had satisfactory results for the management (Haematobia irritans) (Diptera: Muscidae) on of skdr and kdr resistance alleles. Beef Cattle

Dr. Sonja Swiger1 OA34.02 Cypermethrin Resistance in Stable 1Texas A&M AgriLife Extension, Stephenville, Fly Populations from Central Brazil United States

Dr. ATM Barros1, VD Rodrigues2, Dr. PHD The horn fly, Haematobia irritans (L.) (Diptera: Cançado1, Dr. Luisa Domingues3 Muscidae), is one of the most important pests 1Sanidade Animal, Embrapa Gado de Corte of the beef cattle industry. Horn fly adults - CNPGC, Campo Grande, Brazil, 2Curso de are blood feeders that remain in constant Medicina Veterinária, Universidade Católica contact with cattle, providing management Dom Bosco - UCDB, Campo Grande, Brazil, opportunities via insecticide-impregnated ear 3USDA-ARS Knipling Buschland Livestock tags. Controlling horn flies in the United States Insects Research Laboratory, Kerrville, United is time consuming and costly, but failure to States implement management can lead to weight loss and decreased weight gain of calves and In the last decade, large-scale proliferation yearlings. In the past decade, new chemical of stable fly (Stomoxys calcitrans) (Diptera: combinations have been impregnated into ear Muscidae) in organic residues and byproducts tags for pest management. The objectives of of ethanol production has become an this project were to 1) evaluate the efficacy unprecedented problem for cattle producers in of ear tags against horn fly populations and some locations of the Southeast and Midwest 2) determine if reduced fly density results regions of Brazil. As with other livestock in economic return. Several years of data pests, insecticides have been commonly compiled by insecticide class show significant used by ranchers to control this pest in cattle reductions in horn fly populations with the use herds nearby sugarcane mills. However, there of macrocyclic lactone treatments, pyrethroid is limited knowledge about the effects of treatments, and organophosphate treatments insecticides on immature and adult stages in compared with untreated animals. In Texas, the field, and almost no information on the this reduction has been shown to be more susceptibility of stable fly populations to the effective when tagging of cattle is delayed. insecticides commercially available. The present study aimed to evaluate the susceptibility to cypermethrin of two S. calcitrans colonies kept at Embrapa Beef Cattle (Campo Grande,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 126 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA34.04 Examining Cattle Producer in morphological features that it has defied Management of Horn Flies, Haematobia separation. In certain areas such as eastern Irritans (L.) (Diptera: Muscidae) Tennessee, typical T. sulcifrons fly distinctly earlier (June to August) than the darker R. T. Trout Fryxell1, K. Lewis1, S. Schexnayder1, form, which flies later (August to October). A. P. Griffith1, D. B. Taylor2, P. Olafson2, L. Dual seasonal peaks are suspicious for a McKay1 group that generally is univoltine. We have 1University Of Tennessee, 2505 EJ Chapman done morphological, molecular (16S and Dr, United States, 2USDA-ARS COI mitochondrial genes), and quantitative morphometric studies using T. sulcifrons s.l. Horn flies (Haematobia irritans (L.)) are not from across a wide geographic range. The only an economic concern to cattle producers, late-flying and darker form, also called the but also a health and welfare concern because “Carolina form” by some tabanid taxonomists, of their blood-feeding behaviors. Economic is indeed distinct from typical T. sulcifrons. losses to the cattle and dairy industries from The range of the late form extends from this obligate behavior include decreased southern New Jersey to northern Florida, and weight gain, loss in milk productivity, and west at least to about western Tennessee. transmission of bacteria causing mastitis in The late-flying form is the most abundant cattle. Horn fly management strategies are large horse fly attacking livestock in much labor intensive and can become ineffective of the mid-South (North and South Carolina, due to the horn fly’s ability to develop Georgia, eastern and central Tennessee, and insecticide resistance. Tennessee and Texas Alabama). The taxonomic situation in the cow-calf producers were surveyed to examine western portion of their apparent range is cattle producer management of horn flies. less clear, and ongoing studies are addressing While consumer surveys are prevalent in that. After much detective work in the old agricultural economics literature, producer literature, and despite heavy damage on surveys are less common given a producer the type specimen, the late-flying species panel is less assessable than a consumer is probably T. variegatus Fabricius 1805. panel. This paper will describe the results and That species name has not been used since the potential for introducing novel horn fly a Tabanus revision in 1938. We will present management strategies. the scientific and historical evidence for this conclusion, and a paper resurrecting and redescribing T. variegatus is in preparation. OA34.05 Back From the Dead: Resurrection of the USA Horse Fly Tabanus Variegatus Fabricius 1805 OA34.06 Adult Population Dynamics of the Stable Fly (Stomoxys Calcitrans, L.) on Dairy Bradley Mullens1, Rebecca Fryxell2, Paul Farms of Manitoba, Canada Masonick1, Travis Davis2 1University of California, Riverside, United Ms. Gina Karam1, Dr. Kateryn Rochon1 States, 2University of Tennessee, Knoxville, 1University of Manitoba, Winnipeg, Canada United States The stable fly (Stomoxys calcitrans L.) is one Tabanidae (horse and deer flies) are major of the most important livestock pests in North pests of livestock and sometimes people. America. Fly bites are painful and host energy The blood-feeding North American tabanid is diverted to avoidance behaviours, reducing fauna is well known taxonomically overall. weight gain and decreasing milk yields in However, the highly variable species Tabanus dairy cattle. Environmental conditions such sulcifrons Macquardt, which is widespread as temperature, precipitation and substrate and abundant in the eastern USA, has been suitability vary throughout the duration of the difficult to resolve using morphology alone. fly season and affect the number of stable In the southern USA a slightly smaller and flies that successfully emerge and reproduce. darker-colored variant has been suspected of Six Coroplast® sticky traps were deployed being different, but there is enough overlap weekly at three dairy farms from June 17 to

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 127 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

October 21 in 2017 (n=53,540 flies) and May 5Laboratorio de Inmunología, Facultad de 23 to October 3 in 2018 (n=42,585). In 2017, Ciencias Veterinarias, Universidad Nacional de stable flies were first trapped on June 17 and Cajamarca, Perú, Cajamarca, Perú, 6School of population distribution was unimodal with the Biological Sciences, The Queen´s University highest population recorded between July of Belfast, Belfast, United Kingdom, 7Instituto 14-27. In 2018, stable flies were first captured de Ganadería de Montaña (CSIC-Universidad on June 6 and population distribution was de León), Department of Animal Health, bimodal with peaks in July 24-August 2 Grulleros, Spain and August 30-September 6. The sex ratio was determined and used to uncover any The increasing widespread development of trap biases. Females removed from sticky drug resistance in the liver fluke Fasciola traps were dissected to determine ovarian hepatica has motivated the need for development (stage 0-4, nulliparous and alternative diagnostic tools. The work uniparous), which was used to determine reported here describes the validation of an changes in the population age structure egg hatch test (EHT) as an in vitro technique throughout the season. Relationships between to detect albendazole (ABZ) resistance in F. adult stable fly abundance as functions of hepatica. The validation includes the intra- environmental conditions over time were assay, inter-assay and intra-herd variations, analyzed using multiple linear regression and the comparison of results obtained after models and ANOVA F-tests, revealing relative performing the EHT and a controlled efficacy humidity, maximum air temperature, or soil test. Additionally, the development of the temperature to be the best predictors of fly protocol included the adjustment of different abundance, but the parameters changed critical factors to improve the simplicity of the between years. Very few studies on stable assay. The greatest uniformity between results fly biology have occurred in in Manitoba. within the assay and over time until 8 weeks Knowledge of stable fly population dynamics after gallbladder eggs collection (the deadline provides critical information on the timing of proposed for egg analysis), was obtained after life events linked to environmental conditions, incubation with an ABZ concentration of 0.5 and can aid in predicting outbreak patterns µM. The length of exposure to ABZ was shown and lead to strategic management plans. to be critical, as prolonged (15 days) ABZ incubation led to a reversal of drug resistance. There was a close agreement between the OA35 Ruminant Trematodes I outcome of the EHT and that obtained for the in vivo assays. Moreover, the same level July 10, 2019, 11:00 - 12:30 of resistance was observed when eggs and Breakout Room 5, Meeting Rooms KLOP, Level 4 faeces were collected from animals of four (4) different farms and analyzed with both the EHT and the faecal egg count reduction OA35.01 Diagnosis of Albendazole test. A 0.5 µM drug level is confirmed as the Resistance in Fasciola Hepatica discriminating concentration to predict ABZ resistance by the EHT in F. hepatica. Dr. Luis Alvarez1, Dr. Laura Ceballos1, Dr. Candela Canton1, Dr. Cesar Pruzzo2, Dr. Rodrigo Sanabria3, Dr. Laura Moreno1, Dr. OA35.02 On Farm Risk Mapping of Liver Jaime Sanchis4, Prof. Pedro Ortiz5, Prof. Ian Fluke (Fasciola Hepatica): Current Evidence Fairweather6, Prof. Carlos Lanusse1, Dr. Maria and Future Directions Martinez Valladares7 1Centro de Investigación Veterinaria de Tandil Dominique Maree Marendy1,2, Derek (CIVETAN), UNCPBA-CICPBA-CONICET, Schneider1, Dr. Lillian Mukandiwa1, Dr Tommy Tandil, Argentina, 2Facultad de Ciencias L.F. Leung1, Dr. Leslie Gabor2, Dr. Emma K Veterinarias, UNLP, La Plata, Argentina, Doyle1 3INTECH, CONICET-UNSAM, Chascomús, 1School of Rural and Environmental Science, Argentina, 4Departamento de Parasitología, University of New England, Armidale, Universidad de la República, Salto, Uruguay, Australia, 2Elanco, West Ryde, Australia,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 128 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

3School of Science and Technology; Precision management on property. These results Agriculture Research Group, University of are discussed as well as future directions New England, Armidale, Australia to improve the accuracy of this mapping technique. Fasciolosis is a disease affecting approximately 50 countries worldwide and resulting in an estimated US$3.2 Billion OA35.03 Evaluation of Anthelmintic Efficacy due to loss of production. Distribution and of Silver Nanoparticles against Fascciola transmission of liver fluke is dependent on Hepatica, Parasite of Ruminants, Lahore, the presence of lymnaea, the intermediate Pakistan host, which is limited to particular geographic regions. The snail and the free-living stages Dr. Asma Abdul latif1 of the parasite require narrow environmental 1Department Of Zoology,lahore College For conditions to survive. Geographic information Women University, Lahore, Pakistan system (GIS) mapping provides a unique tool to identify high risk grazing areas for Parasitic diseases affect lots of people world stock infection. While GIS mapping has been widely, particularly in developing countries. validated at a regional level, it has not yet Fasciola hepatica infects humans as well as been done at a farm level. ruminants, it causes Fasciolosis and it is also worldwide in distribution. Fasciola hepatica The initial study investigated potential is prevalent in different areas of Pakistan in indicators of metacercariae presence, on a ruminants. Synthetic anthelmintics/Vaccines commercial sheep and cattle property in the are not effective for the control of Fasciolosis New England region of New South Wales, due to parasitic resistance, therefore, new Australia. Measurements utilised included therapies are being explored for the control an electromagnetic induction survey to of parasites. Various Nano particles are being ascertain soil apparent electrical conductivity examined on wide scale as antiparasitic drugs. (ECa), a proximal normalised difference The objective of the present study was to vegetation index (NDVI) survey and a digital evaluate the anthelmintic activity of silver elevation model (DEM) was created using nanoparticles against Fasciola hepatica. Adult a differentially corrected geographical Fasciola hepatica were isolated from 100 liver positioning system (dGPS). samples of freshly slaughtered ruminants. The efficacy of seven different concentrations The sampling locations were randomly (mg/ml) of silver nanoparticles from 0.17, selected from 3 management zones created 0.085, 0.043, 0.021, 0.011, 0.0053 to 0.0027on by interpolating the three spatially measured adult flukes of Fasciola hepatica was checked parameters (ECa, NDVI and Elevation) in adult motility Assay. Albendazole drug (12 individually and applying an unsupervised mg/ml) and PBS were used as positive and K-means clustering algorithm. Metacercariae negative control respectively. It was revealed counts were coupled with static ECa, NDVI that the degree of immobilization got delayed and elevation measurements for calibration as the concentrations of silver nanoparticles purposes at these sample sites. Metacercariae were reduced. At highest concentration of density was positively associated with pH silver nanoparticles the mean time (min) (P<0.0001), ECa (p=0.0109) and NDVI for paralysis and mortality was recorded as (p=0.0039). Examination of the cluster 1.67±0.234 and 1.83±0.24 respectively. At means showed that one of the identified lowest concentration of silver nanoparticles management zones aligned directly with 0.0027mg/ml mean time for paralysis and the hypothesised higher risk parameters. death of flukes was recorded as 19.33±1.41and Overall, this one-off scoping study suggests 21.67±1.25 respectively. Total time recorded that this management zoning approach for death and paralysis of Fasciola hepatica and conversion, with ground-truthing, to a flukes in positive control of Albendazole “risk map” could be implemented as a tool 12mg/ml drug was recorded as 8.56±0.16 and to improve sampling programs for research 6.77±0.32 respectively. The Present study purposes and guide site-specific parasite showed substantial anti-parasitic effect of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 129 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

silver nanoparticles on adult flukes of Fasciola to Australian conditions requires further hepatica and hence emerged as dynamic and investigation. effective drug with minimum side effect as compared to other drugs. 1. George et al. (2017) Veterinary Parasitology,246, 60-69. 2. George et al. (2019) Vet Parasitology: X, OA35.04 Assessment of Factors Influencing https://doi.org/10.1016/j.vpoa.2018.100001 Faecal Diagnostic Outcomes and Predictive Modelling of Endemic Fasciola Hepatica on Australian Sheep and Cattle Farms OA35.05 Fasciola Hepatica: Secreted Molecules Associated With Liver Sarah George1, Ashley George1, Dr Peter Pathogenesis Rolfe1, Prof David Emery2 1Elanco Animal Health, Kemps Creek, Australia, Dr. Krystyna Cwiklinski1,2, Heather Jewhurst1,2, 2University of Sydney, Camden, Australia Dr. Oksana Lyubomska1, Orla Drysdale1, Dr. Carolina De Marco Verissimo1,2, Dr. Mark The diagnosis, monitoring and flukicide Robinson1, Dr. Sheila Donnelly3, Prof. John efficacy testing of fasciolosis on-farm is reliant Dalton1,2 on non-terminal methods. The coproantigen 1Queen’s University Belfast, Belfast, United ELISA (cELISA) has been recommended Kingdom, 2National University of Ireland for diagnosis of fasciolosis and associated Galway, Galway, Ireland, 3University of flukicide efficacy testing as an alternative Technology, Sydney, Sydney, Australia to fluke egg counts (FFEC) for monitoring parasitism. The liver fluke Fasciola hepatica is an economically important worm pathogen Experimental multi-age F.hepatica infection in of humans and their livestock worldwide. a controlled study1 and subsequent monthly Infection of the mammalian host involves monitoring on endemic sheep and cattle farms parasite activation from an encysted stage over 12 months2 indicated greatest diagnostic followed by penetration through the intestinal sensitivity when cELISA and FFEC were utilised wall and migration to the liver. The major in parallel. To analyse factors influencing pathogenesis associated with infection diagnostic outcomes on endemic farms linear results from the extensive liver damage models were applied for each diagnostic alone caused by the tunnelling and feeding activity and as binary data, both in parallel and series. of migrating immature flukes coupled with Variables examined were of climatic origin, the pathology associated with host immune locality, treatment status and species with fixed responses. Within the liver, the parasite’s effects being sample time and individual animal growth advances rapidly, doubling in size included as a random effect. Additionally, approximately every two weeks, alongside the suitability of the Ollerenshaw Index as a the development of the digestive and predictive model for fasciolosis under Australian reproductive structures. Here, we report conditions was examined. our transcriptomic and proteomic approach to determine the factors associated with Linear regression models demonstrated parasite development and the resulting the results of the two diagnostics utilized liver pathogenesis. Investigation of gene in parallel were explained substantially (R2= transcription demonstrated that the immature 0.91) as were series data (R2 =0.88) when the flukes up-regulates >8000 transcripts respective models were fitted. In contrast, once within the liver. Specifically, genes the fitted models for FFEC (R2=0.54) and associated with liver fluke metabolism and cELISA (R2=0.58) were poor explanations the mechanisms associated with active for test outcomes. The outcomes of these feeding and immune evasion, were found models support previous findings that to be enriched. These analyses were further suggest that the two diagnostic tests are corroborated by proteomic analyses, which best utilized together, particularly in parallel. highlighted that the parasites secrete a The application of the Ollerenshaw Index plethora of proteins to negotiate the range

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 130 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

of different host molecules, tissues and lifecycle were elucidated. Particularly, we micro-environments they will encounter and found an abundance of transcripts associated to effect the degradation of the liver tissue. with the immune response for both hosts, This study provides information vital to indicating that the parasite manipulates our understanding of fluke/host interactive the immune system of snail and mammal biology that can be exploited for the to ensure its own survival. 70% of the F. development of control strategies that target gigantica transcripts share homology with F. this pathogenic stage. hepatica genes; however, the profile of the abundantly expressed transcripts within the various lifecycle stages (egg, metacercariae OA35.06 Dynamic Transcriptomic Analyses and adult) imply species-specific regulation of the Entire Life Cycle of Fasciola Gigantica that could reveal specific adaptations that Identify Key Processes Through Vastly have taken place since these two parasites Different Environments diverged 25 million years ago. This is the first analysis of the transcriptional profile of Dr. Xiao-Xuan Zhang1, Dr. Krystyna Cwiklinski2, the F. gigantica lifecycle, which identified Mr. Rui-Si Hu1, Mr. Wen-Bin Zheng1, Mr. Zhao- key processes the parasite must undergo An Sheng3, Mr. Fu-Kai Zhang1, Dr. Hany as it progresses through vastly different Elsheikha4, Professor John Dalton5, Professor environments. The comparative analysis with Xing-Quan Zhu1 F. hepatica has provided further insight into 1Lanzhou Veterinary Research Institute, Fasciola biology, crucial for the development Chinese Academy of Agricultural Sciences, of novel control strategies against both Lanzhu, China, 2Queen’s University Belfast, Fasiola species. Belfast, United Kingdom, 3Guangxi University, Nanning, China, 4University of Nottingham, Loughborough, United Kingdom, 5National OA36 Treatment and Control of GI University of Ireland Galway, Galway, Ireland Nematodes in Ruminants

Fasciola gigantica is a digenean trematode July 10, 2019, 13:30 - 15:30 and causative agent of fasciolosis, an Plenary Hall, Madison Ballroom (ABCD), Level 4 economically important livestock disease and zoonosis. In this study, we reveal the dynamic transcriptional changes that occur throughout OA36.01 Performance Characteristics the parasite lifecycle. A total of 58,422 of the Adequacy of Infection Criteria transcript clusters (unigenes) were assembled Recommended in VICH GL 7 for Anthelmintic and re-mapped to the eight transcriptomes Efficacy studies representing the various lifecycle stages. Dr. Xiongce Zhao1, Dr. Emily Smith1, Dr. 7445 unigenes were transcribed by all 1 1 the lifecycle stages, with the remaining Aimee’ Philippi-Taylor , Dr. Virginia Recta 1FDA/Center For Veterinary Medicine, unigenes (n=50,977) exhibiting stage- Rockville, United States specific expression. The free-living miracidia transcribe a myriad of genes indicating this stage is prepared to seek out and invade Veterinary International Committee for the snail intermediate host. Consistent with Harmonization Guideline 7 (VICH GL7) “Efficacy previous studies of F. hepatica metacercariae, of Anthelmintics: General Requirements” F. gigantica metacercariae are metabolically recommends criteria for adequacy of infection active, though they display up-regulation in anthelmintic studies. The criteria include the of transcripts involved in regulating gene option to use statistical confidence limits based expression. By parsing the data derived from on the geometric mean or the median worm the stages that infect and develop within the burden in the control group. snail (miracidia, rediae and cercariae) with the buffalo-specific stages (juvenile 42dpi, Using simulation studies, we investigated the juvenile 70dpi and adult parasites), molecules performance of the statistical criterion given that are important for each stage of the in VICH GL7 for determining the adequacy of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 131 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

infection in anthelmintic studies. The results determine the average daily gain and feed show that under certain circumstances the efficiency. Carcass quality information was statistical criterion is useful in safeguarding obtained at slaughter. Pens were considered against overestimating the adequacy of the experimental unit. Anthelmintic treatment infection as the sample size increases. had no effect on the average daily gain and However, the statistical criterion may be feed efficiency during backgrounding or overly restrictive for samples with zero counts finishing. However, compared to the control and may not protect against overestimating group, some superior carcass quality traits adequacy of infection when the sample (quality grade, yield grade, and marbling contains no zero counts. score) were significantly more frequent in anthelmintic treated calves. was the predominant parasite species in all OA36.02 Effects of Anthelmintic Treatment calves before anthelmintic treatment혖혴혵혦혳혵혢혨혪� and혰혴혵혦혳혵혢혨혪 on Production Performance, Carcass Quality, in control calves 14 days post-treatment. and Predominant Nematode Species in After treatment, was the Feedlot Cattle from Western Canada predominant parasite species in ivermectin- treated calves while혊혰혰혱혦혳혪 no parasite� 혰혯혤혰혱혩혰혳혢 eggs were Mr. Eranga De Seram1, Dr. Gregory Penner1, recovered from calves given the combined Dr. John Gilleard2, Dr. John Campbell1, Dr. treatment. Anthelmintic treatment improved Elisabeth Redman2, Dr. Colleen Pollock3, Mr. some carcass quality traits in these calves, Samantha Ekanayake1, Dr. Fabienne Uehlinger1 the economic significance of which remains 1University of Saskatchewan, Saskatoon, to be determined. in Canada, 2University of Calgary, Calgary, control calves may suppress superior carcass Canada, 3Merck Animal Health, Kirkland, characteristics and this혖혴혵혦혳혵혢혨혪 requires� 혰혴혵혦혳혵혢혨혪 further research. Canada

Changes in climate and anthelmintic OA36.03 Relative Effectiveness of a Slow susceptibility are changing the epidemiology Release Drug Versus Strategic Administration of gastrointestinal nematodes in livestock. of a Short Acting Anthelmintic for However, the production impact of Controlling Nematodes in US Beef Cattle gastrointestinal nematodes in beef cattle from western Canada has not been studied Dr. Louis Gasbarre1, Dr. William Epperson2, in decades. We determined the effects of Dr. Dante Zarlenga3, Dr. Paul Beck4, Dr. Harold currently used anthelmintics on production Newcomb5 performance, carcass quality characteristics, 1Gasbarre Consulting, Buffalo, United States, and species diversity in western Canadian 2CVM Mississippi State university, Starkville, feedlot calves. A randomized controlled trial United States, 3Parasitic Disease Laboratory, was conducted with 234 auction market- ARS, USDA, Beltsville, United States, 4Dept derived, weaned, fall-placed steer calves. Animal and Food Sciences, Oklahoma State Calves were assigned to three treatment University, Stillwater, United States, 5Merck groups: control; injectable ivermectin; Animal Health, Kenilworth, United States combination of injectable ivermectin and oral fenbendazole. Each group contained There has been an increase in the use of a replicates of 6 pens with 13 animals per slow release long acting macrocyclic lactone pen. Calves were treated according to in beef cattle operations in the United States. individual body weights and manufacturers’ Given the potential for development of specifications. Individual fecal samples were anthelmintic resistance in such a program, collected to determine the pre- and post- a study was designed to ascertain if older treatment fecal egg counts by modified strategic anthelmintic programs using a Wisconsin sugar flotation. The predominant product with no residual activity could be parasite species in each treatment group as effective on parasite control and animal was determined using a deep-sequencing productivity. Two hundred calves were nemabiome assay. Monthly body weights acquired from sale barns throughout the and pen level feed intake were used to southeastern US. Calves were divided into

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 132 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

two groups; one receiving slow release lambing has been suggested as one factor eprinomectin as per label instructions that may increase selection for MOX (Group 1) while the other received an initial resistance in gastrointestinal nematodes. combination treatment with oral fenbendazole Recommendations to counter this potential and injectable ivermectin followed by a problem suggest that the use of a short topdressed mineral containing fenbendazole acting anthelmintic (exit-drench) at the in the daily feed supplement on days 28- end of the persistent period of activity can 31 and 56-59 (Group 2). Calves from each remove survivors from the MOX treatment group were slaughtered and parasites were and reduce the selection pressure for MOX recovered, enumerated, and identified to resistance, but no empirical data exists species at days 10, 66, and 114. Total mean to back up these recommendations in worm recoveries day 10 were: Group 1 - 5,247 Temperate Europe. A replicated field trial was worms consisting of Cooperia punctata (Cp) undertaken over the grazing seasons of 2017 and C. oncophora (Co), Haemonchus sp and 2018 assessing the impact of different (Hae), and Oesophagostomum radiatium (Or); treatment protocols on lamb productivity Group 2 – 3 larvae in the abomasum. At day and parasitological parameters (faceal egg 66: Group 1 - 16,973 worms – Cp, Co, Hae, Or, count, species composition). Ewes with twin and Ostertagia ostertagi (Oo); Group 2 - 40 lambs were grazed on 12 paddocks (n=5 and worms – Oo and larval Cooperia. At day 114: 10 respectively) between spring and late Group 1 - 7,737 worms and Group 2 - 3,532 autumn. Ewes were treated at turn out with worms, with all parasite species previously MOX (Cydectin 0.1% Oral drench)±monepantel identified in both groups. A representative (Zolvix; 5 weeks post MOX), lambs were sample of recovered Hae were subjected to treated at weaning and mid/late season PCR and all were shown to be H. placei. There with either MOX or ivermectin (Oramec) was no significant difference in weight gains or moxidectin+monepantel (as above) at between the 2 groups at the termination weaning. Results at the end of the trials on the experiment on day 123. Previous showed that the difference between the studies have indicated the Cooperia sp are poorest responding group (MOX administered increasingly surviving macrocyclic lactone to ewes and lambs) and the other treatment treatments. This study indicates that this may strategies were; Live-weight gain differences now be true of H. placei and O radiatum. were between 3-6Kg heavier over 140 days and worm burdens were greater 2x lower. Moxidectin efficacy based on worm burdens OA36.04 Longitudinal Study on the Effect ranged between 98%-100% and 85%-100% for of Moxidectin Administration With and years 2017 and 2018 respectively. The results Without an ‘Exit’ Drench on Production and demonstrate that an exit drench strategy may Parasitological Parameters in a UK Sheep have a positive impact, in the short-term, Flock when administering moxidectin.

Dave Bartley1, Leigh Andrews1, Arundhati Rao1, Alison Morrison1 OA36.05 Anthelmintic Efficacy and 1Moredun Research Institute, Penicuik, United Pharmacokinetics of Topical 0.5% W/V Kingdom Eprinomectin (EPRINEX®) Administered at 1 Mg per Kg Body Weight in Lactating Dairy Long-acting anthelmintics, like moxidectin Goats With Induced Nematode Infections (MOX), are powerful endectocides used for the treatment of both gastro-intestinal Dr. Dietmar Hamel1, Dr. Steffen Rehbein1, nematodes and sheep scab mites. Recent Valerie Kvaternick2, Dr. Hailun Wang3, Dr. studies in the UK suggest an increase in Michael Kellermann1, Sandra Mayr1, Renate MOX resistant nematodes. Due to the Rauh1, Martin Visser1, Thea Wiefel1, Becky perceived production benefits afforded Fankhauser3 to lambs later in the grazing season many 1Boehringer Ingelheim Vetmedica GmbH, UK farmers drench ewes with MOX around Kathrinenhof Research Center, Rohrdorf, lambing. The use of MOX in ewes around Germany, 2Boehringer Ingelheim Animal

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 133 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Health USA, Inc., North Brunswick, NJ, United lactating dairy goats was highly efficacious States, 3Boehringer Ingelheim Animal Health against gastrointestinal and pulmonary USA, Inc., Duluth, GA, United States nematode infections. These results as well as the pharmacokinetics in lactating dairy goats INTRODUCTION: Lactation is discussed of eprinomectin are similar to that observed in as a physiological covariate which may young growing and adult female dry goats. influence the exposure characteristics of systemically acting drugs including macrocyclic lactones and potentially alter OA36.06 Nemabiome Analysis Reveal a their pharmacological response. Therefore, Change in Parasite Fauna After Years of efficacy and pharmacokinetics of topical 0.5% Anthelmintic Treatment w/v eprinomectin, which has been recently authorized in Europe as EPRINEX® Multi as a Dr. Peter Halvarsson1, Professor Johan broad-spectrum anthelmintic with zero hours Höglund1 milk withdrawal in sheep and goats, were 1Swedish University of Agricultural Sciences, evaluated in a GCP and VICH anthelmintic Uppsala, Sweden testing guidelines compliant, blinded study using lactating dairy goats with induced Parasitic nematode infections are abundant in nematode infections. grazing livestock and they put a constraint in maintaining animal health and welfare, being METHODOLOGY: Twenty lactating German the most important factor that limits livestock White Noble goats, harboring induced production. Since the advent of anthelmintic infections of adult gastrointestinal nematodes drugs, heavy infections can be treated and lungworms were ranked on pre-treatment effectively. Recently, a buildup of resistance to bodyweight and allocated at random either anthelmintics has been observed all over the to remain untreated (control) or to be world. In this study we are investigating the treated with EPRINEX® topically at 1 mL per effects of anthelmintic treatments on parasitic 5 kg body weight. Plasma concentrations of nematode communities in 35 Swedish sheep eprinomectin B1a were determined in blood farms between 2007 and 2016. The first cases samples collected prior to treatment and at of anthelmintic resistance (to Valbazen) were specific times up to necropsy. A necropsy reported in 2007 and recommendations for was performed 14 days after treatment and treatment changed to Ivermectin. However, nematode counts made to determine efficacy. Ivermectin resistant Haemoncus contortus are now present on many farms. Larval fecal RESULTS: Treatment was well accepted cultures from 50 samples were collected and no health problems were observed before and after anthelmintic treatment throughout the study. Counts of adult respectively. The conserved ITS-2 region Dictyocaulus filaria, Haemonchus contortus, of the nematode rRNA was PCR amplified Teladorsagia circumcincta(pinnata/trifurcata), and sequenced using PacBio© SMRT Trichostrongylus axei, T. colubriformis, cell technology. Using this strategy, the Cooperia curticei, Nematodirus battus nemabiome of each sample was characterized and Oesophagostomum venulosum were for the whole nematode community. In the significantly (p<0.0070) lower in the treated period from 2007 to 2016, H. contortus, the goats (>95% reduction relative to controls). nematode considered the most pathogenic for Basic pharmacokinetic parameters of sheep, showed a 100% increase in frequency eprinomectin (B1a component) were: AUClast, of all ITS-2 sequences, while the two other 23.8 ± 9.7 day*ng/mL, Cmax, 5.55 ± 2.27 common species, Teladorsagia circumcincta ng/mL, MRTlast, 3.59±0.60 days, and T1/2, and Chabertia ovina showed 56% and 80% 3.57±0.77 days; individual maximum plasma decrease of the ITS-2 sequences respectively. concentrations were observed from 8 to 48 Furthermore, after treatment, H. contortus hours (median of 12 hours). showed a decreased sensitivity for Ivermectin and Valbazen. In contrast, treatment with In conclusion, eprinomectin administered Levamisole remained effective against H. topically at 1 mg per kg body weight to contortus on the investigated farms.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 134 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA36.07 Boron-containing Compounds Are OA36.08 New Benzimidazole Derivates As Rich Source of Future Antiparasitic Agents Potential Anthelmintic Compounds

Yong-Kang Zhang1, Tony Chun Yu Liu1, Elora Valderas1, María Álvarez1, Verónica Yasheen Zhou1, Chunliang Liu1, Marissa Castilla1, Rafael Balaña1, Esther Del Olmo2, Aubrey1, Jacob J. Plattner1 María Martínez1 1Boragen, Inc., Durham, United States 1Universidad de León, León, España, 2Universidad de Salamanca, Salamanca, Finding new chemical entities (NCEs) with España novel mechanisms of action is a common approach to combat parasitic resistance and Gastrointestinal nematode infections improve treatment efficacy. Boron chemistry cause direct and indirect losses of great provides an emerging area of rich NCEs economic impact worldwide by decreasing demonstrating broad antiparasitic activities. productions yields and increasing health For examples, it has been reported that care cost in sheep. Thus, the aim of this boron-containing compounds have excellent study was to test the nematocidal effect of a activities against tick and flea¹,², against new collection of benzimidazole derivatives malaria-causing parasite P. falciparum³, , against Teladorsagia circumcincta by in vitro against GI worms , against Animal African methods. Compounds were tested in eggs, Trypanosomiasis (AAT) parasites T. ⁴ first stage larvae (L1) and thrid stage larvae congolense and T.⁵ vivax , and against Human (L3) by means of the Egg Hatch Assay, L1 African Trypanosomiasis (AAT) parasite T. Mortality Assay and the Larval Migration brucei . This presentation⁶ will provide an Inhibition Assay. An initial screening was done overview of boron chemistry antiparasitic at a single concentration of 50 M; those application⁷ and predict possible future molecules that showed an activity higher than research direction. >90% were selected to to determineµ their half maximal effective concentration (EC50) and References were tested again against a resistant strain. 1. Y.-K. Zhang, J. J. Plattner, E. E. Easom, Y. Finally, the anthelmintic effect was compared Zhou, T. Akama, W. Bu, W. H. White, J. M. with their cytotoxicity in two digestive Defauw, J. R. Winkle, T. W. Balko, et al., Bioorg. established cell lines, Caco-2 and HepG2, and & Med. Chem. Lett., 2015, 25, 5589–5593. their corresponding concentrations required 2. Y.-K. Zhang, J. J. Plattner, E. E. Easom, Y. to reduce cell viability by 50% (CC50) were Zhou, T. Akama, W. Bu, W. H. White, J. M. determined in order to establish a selective Defauw, J. R. Winkle, T. W. Balko, et al., Bioorg. index (SI) for each molecules. Only 5 of the 18 & Med. Chem. Lett., 2016, 26, 3182–3186. compounds were able to arrest the hatching 3. Y.-K. Zhang, J. J. Plattner, E. E. Easom, R. of eggs in the susceptible strain in more than T. Jacobs, D. Guo, V. Sanders, Y. R. Freund, B. 99% at 50 M and 4 of them were also able Campo, P. J. Rosenthal, W. Bu, F.-J. Gamo, L. to produce a significant effect on the resistant M. Sanz, et al., J. Med. Chem., 2015, 58, 5344- strain used μin the current study. Considering 5354. only those compounds that have a significant 4. Y.-K. Zhang, J. J. Plattner, E. E. Easom, R. T. effect, their CC50 values were very Jacobs, D. Guo, et al., J. Med. Chem., 2017, 60, homogeneous between Caco-2 and HepG2, 5889-5908. which makes that SI values were comprised 5. T. Akama, K. Jarnagin, J.J. Plattner, S.R. between 1.84 to 3.60 for Caco-2 cells and Pulley, W.H. White, Y.K. Zhang, Y. Zhou, WO 1.53 and 2.73 for HepG2 cells. These results 2014149793. indicate that they could be good candidates 6. T. Akama, Y.-K. Zhang, Y. R. Freund, P. Berry, for further studies to determine their in vivo J. Lee, E. E. Easom, R. T. Jacobs, J. J. Plattner, activity. Study funded by AGL2016-79813-C2- M. J. Witty, et al., Bioorg. & Med. Chem. Lett., 1-R, Junta de Castilla y León co-financed by 2018, 28, 6–10. FEDER [LE020P17], EVG by FPU17/00627 and 7. Y.-K. Zhang, Y. R. Freund and R. T. Jacobs, MMV by RYC-2015-18368. Comprehensive Medicinal Chemistry III, volume 8 (2017).

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 135 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA37 Poultry Coccidia, Aquatic cholesterol, total protein and A/G ratio with Infections increases in AST, ALT, ALP and GGT activities and creatinine, uric acid, and total globulins July 10, 2019, 13:30 - 15:30 levels. Histopathological examination revealed Breakout Room 2, Hall of Ideas E&H, Level 4 in the (G2) post challenge; severe changes in the liver, gall bladder, spleen, intestine and kidney. While, less histopathological OA37.01 Hemato-Biochemical and changes were recorded in the (G3) and (G4) Histopathological Investigations Following and non-significant changes were recorded Concurrent Coccidia and Clostridia in the (G5). In conclusion, using the Coccidia Experimental Infection in Broiler Chickens vaccine beside Clostridia toxoid minimized Vaccinated With Clostridia Toxoid and/or the severity in hematological and biochemical Coccidia Vaccine parameters and histopathology of the internal organs in infected chicks. Dr. Ahmed El-Shemy1, Professor Safaa Yassin2, Professor Hamdy Soufy1, Professor Alaa 2 1 Ahmed , Professor Soad Nasr OA37.02 The Role of Parasitic Crustacea as 1 Department of Parasitology and Animal Vectors of Aquatic Diseases Diseases, National Research Center, 33 Bohouth Street, Dokki, P.O. Box 12622, Giza, Dr. Nico J Smit1, Dr. Kerry A Hadfield1 2 Egypt, Department of Clinical Pathology, 1North-West University, Potchefstroom, South Faculty of Veterinary Medicine, Cairo Africa University, P.O. Box 12211, Giza, Egypt In assessing the role of parasitic crustaceans Concurrent infection with Coccidia and as vectors, it is interesting to note that the Clostridia is one of the major diseases relevant literature appears limited to a handful affecting chickens causing economic losses. of species. Isopods of the genus Gnathia This experiment aimed to evaluate the effect (family Gnathiidae) likely act as definitive of concurrent experimental infection with hosts and vectors of fish blood parasites of Eimeria tenella and Clostridium perfringens the genus Haemogregarina. They may also type A -local isolates- on hematological and transmit fish viruses (such as viral erythrocytic biochemical parameters and histopathology necrosis), and may be intermediate hosts for of the internal organs of chicks vaccinated nematode larvae. Furthermore, cymothoid with Clostridia toxoid and/or Coccidia. A isopods (family Cymothoidae) may transmit total of 225 -one day old- broiler chicks were lymphocystis virus to fishes. Recent studies divided into 5 equal groups; (G1): normal show barnacles (subclass Thecostraca) on control, (G2): infected, (G3): vaccinated with the carapace and gill filaments of crabs Coccidia vaccine on the 3rd day old, (G4): could be potential reservoir hosts for shrimp treated with Clostridia toxoid at the 5th viruses. Copepods of the genus Caligus and day old, and (G5): vaccinated with Coccidia Lepeophtheirus (family Caligidae) are noted vaccine on the 3rd day old and treated with as potentially important mechanical vectors Clostridia toxoid at the 5th day old. All groups or alternative hosts of a number of viral were challenged orally after 21 days post diseases between fishes and Lepeophtheirus vaccination (dpv) with Coccidia oocysts at can transfer pathogenic bacteria between a dose of 5×104 and then with Clostridium fishes. Ergasilids (family Ergasilidae) parasitic perfringens type A at a dose of 0.5 ml of 24 h on the gill filaments of fishes can support the cooked meat broth 1×107 CFU at the 23rd dpv. replication of shrimp viruses, and likely act Blood samples and tissue specimens from as viral vectors and transmit lymphocystis. liver, spleen, gall bladder, intestine and kidney Branchiurans, specifically from the genus were collected from all groups at the 5th, 13th Argulus, are thought to serve as mechanical and the 21st dpv and at the 5th, 7th, 15th and vectors of several viruses to fishes, especially the 21st day post challenge. Results revealed carp, as well as acting as intermediate hosts that (G2) showed anemia, leukopenia, for dracunculoid and skrjabillanid nematodes decrease levels of iron, glucose, total of fishes. All of these vector examples are

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 136 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

further discussed within the presentation, that highlighted the sporozoites. From these and areas of possible future research are results, it will be possible to work towards identified. the automated diagnosis of these structures. Therefore, studies on the adequacy of TF-Test technique have been conducted (FAPESP OA37.03 Processing of Bovine Faecal PROCESS: 2014/12236-1), to make possible Samples by New Parasitological Technique the computerized identification of these and Perspectives of Automation in the parasites by building an image database that Diagnosis of Cryptosporidium SPP. will serve to train the classifier in the pattern recognition module of the system wich will Sandra Valéria Inácio1, Professor Jancarlo take into account inner structures of oocyst Ferreira Gomes2, Professor Alexandre Xavier and then offer better alternatives to the Falcão2, Saulo Hudson Nery Loiola2, Bianca procedure and reduce interpretation errors Martins dos Santos2 Celso Tetsuo Nagase improving consequently the efficiency in the Suzuki2, Professor Katia Bresciani1 laboratories routine. 1São Paulo State University (UNESP), School of Veterinary Medicine, Araçatuba, Brazil, 2Laboratory of Image Data Science OA37.04 The Effect Of Long-Term Cell- (LIDS), Institute of Computing, University of Culture Passage On Sarcocystis Neurona Campinas (UNICAMP), Campinas, Brazil Genome Content And Predicting Genes Necessary For Completion Of The Parasite Cryptosporidiosis arouses great interest in Life Cycle the scientific community due to its significant zoonotic potential. The usual techniques of Jamie Norris1, Dr. Sriveny Dangoudoubiyam1, concentration and permanent staining for the Dr. Daniel Howe1 parasitological diagnosis of Cryptosporidium 1Department of Veterinary Science - University spp. oocysts, present limitations in the of Kentucky, Lexington, United States existing protocols, mainly related to the composition of the chemical reagents, low Sarcocystis neurona is an obligate intracellular productivity, lack of coverage and high costs. parasite that causes equine protozoal In view of these problems, we made possible myeloencephalitis (EPM). The S. neurona a new parasitological technique called strain SN3 was isolated from an EPM horse TF-Test Coccidia, that was evaluated and in 1991 and has been passed in cell culture validated in an intralaboratorial study, aiming almost continuously for nearly 30 years. to identify this protozoan in temporary slides To aid molecular and genetic studies of S. which were assessed under conventional neurona, a reference genome for the SN3 light microscopy. For this purpose, 68 faecal strain was produced in 2013. Next generation samples of calves from the city of Araçatuba, sequencing (Illumina) was used subsequently São Paulo/Brazil, were processed by the new to obtain genomic sequence data from three technique just as centrifuged-sedimentation additional S. neurona isolates that had not with negative staining of malachite green. been passed extensively in cell culture (SN4, The cases were confirmed by Nested-PCR. SN-OT1, SN138). Interestingly, alignment of The Kappa index was used to measure the SN4, SN-OT1, and SN138 sequences to the the strenght of agreement between both SN3 reference genome revealed significant procedures. Thus, we noted the success of genomic regions (up to ~120 kb) that are TF-Test Coccidia for detection of oocysts seemingly absent from the SN3 strain. These of Cryptosporidium spp. which showed genomic regions are largely devoid of gene positivity in 34 samples and almost perfect annotations, partly due to a paucity of agreement (Kappa = 1,000). We considered transcript evidence from all stages of the as advances: the low amount of debris in the parasite’s life cycle. We hypothesize that fecal smear, the speed of preparation of the these regions have been deleted from the SN3 slide with a good concentration of oocysts genome because they contain genes that are without morphological deformation and the deleterious to asexual parasite growth in vitro adjustment of a temporary staining solution but potentially important during other stages

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 137 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

of the parasite’s life cycle (i.e., bradyzoites 120 hour prepatent period; oocyst shedding and/or sporozoites). Gene prediction persisted until 10 days post-inoculation. To models based on the closely-related parasite complement biological data, the complete Toxoplasma gondii will be used to determine mitochondrial genome and partial nuclear whether the deleted regions contain genes 18S rDNA were sequenced. Molecular and not identified in the original annotation effort biological observations confirm that this for the SN3 genome. Putative functions Eimeria species has not been reported from associated with these sequences may provide partridges nor any other galliform bird to insights into the natural lifecycle of S. neurona our knowledge. We suggest that it will and related parasites. need formal description as a new species. Understanding of the biology of this, as yet, unnamed Eimeria sp. will inform development OA37.05 Biological and Molecular of vaccination methodologies designed to Characterization of a Pathogenic Unnamed elicit protective immunity, such as: 1) live Eimeria Species Causing Clinical Coccidiosis oocyst vaccination followed by a carefully in Commercial Chukar Partridge (Alectoris monitored 2-step partial house brooding; and, Chukar) Flocks in Ontario 2) a ‘bioshuttle’ (vaccination/anticoccidial combination). Effective coccidiosis control will Jessica Rotolo1, Dr. John Barta1, Rachel Imai1, enhance flock health and increase profitability Perryn Kruth1, Ryan Snyder1 for commercial chukar producers. 1University Of Guelph, Guelph, Canada

A local commercial producer of Chukar OA37.06 When Minor Species Are Not So partridge (Alectoris chukar) has had recurrent Minor: Eimeria Innocua, Far From Innocuous issues with frequent clinical coccidiosis and high flock mortalities. Traditional live Rachel Imai1, Ryan Snyder1, Jessica Rotolo1, Dr. vaccination using a few infective oocysts John Barta1 given to day-of-age birds to elicit protective 1Department of Pathobiology, Ontario immunity has been unsuccessful. Partridge Veterinary College, University of Guelph, chicks are apparently unable to acquire Guelph, Canada protective immunity fast enough to prevent parasite replication and intestinal tract Historically, only Eimeria meleagrimitis and damage. In preparation for exploring alternate Eimeria adenoeides were considered highly methods of vaccination for coccidiosis pathogenic to turkeys. Recently, all other management in commercial Chukar partridge Eimeria spp. that infect turkeys (Eimeria flocks, the responsible pathogenic Eimeria dispersa, Eimeria gallopavonis, Eimeria species was isolated and characterized. The innocua and Eimeria meleagridis) were shown morphology and life cycle of this Eimeria to be present within flocks in commercial species was examined in vivo and sequence- turkey facilities. Detection of these ‘minor’ based genotyped at two genetic loci. species has only become possible recently Morphometrics of oocysts and sporocysts through application of a nested, species were measured using light microscopy with specific PCR assay that can differentiate all computerized image analysis. Experimental 6 species in fecal samples. The last of the infections with coccidia free chukar species added to this assay was Eimeria partridges were used to describe the innocua; this parasite was considered complete endogenous development and relatively non-pathogenic but preliminary daily fecal collection post inoculation was research suggested that this may not be used to determine the prepatent period the case. To clarify the pathogenic potential and duration of shedding. Endogenous of E. innocua, an in vivo infection trial was development was determined histologically conducted. Hens were reared coccidia-free from samples collected at 8 locations along and infected at 14, 23, 30 or 40 days-of- the intestinal tract every 8 hours throughout age. Poults were inoculated by oral gavage prepatency. The parasite had 5 asexual with 100 to 1,000,000 oocysts per bird. Five generations prior to oocyst formation over its days post-inoculation (DPI), turkeys were

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 138 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

necropsied to permit description of the larval cyathostominosis, a potentially macroscopic lesions at the various doses. fatal condition. Other, putative effects of Tissue samples were taken from each bird cyathostomin infection on horse productivity at fixed positions for histopathology. Body and performance are largely unsubstantiated. weights of each bird were taken immediately Regardless, the compulsion to deworm is before inoculation and again at necropsy. sacrosanct among horse owners, even if Eimeria innocua produced macroscopically implementation is based more on tradition obvious pathological changes that, at and myth than scientific evidence. higher doses, extended from the duodenal loop into the ileum (20cm or more beyond To date, effective cyathostomin control Meckel’s). Bleaching, ballooning and thinning measures share the common strategy of of the intestinal mucosa were evident; reducing environmental contamination with unsurprisingly, lesion severity and length eggs. However, application of this strategy is of affected intestinal tract both increased complicated by numerous practical questions, with higher numbers of oocysts inoculated. including which horses to treat, what drugs to However, dose-dependent increases in lesion use, and timing of administration in relation severity were not consistent for the youngest to seasonal and management factors. These poults; maturation of the turkey gut could persistent questions will remain unresolved potentially be a mitigating factor affecting the without a better understanding of the impact of infections and must be considered regulatory mechanisms of cyathostomin when challenge experiments are conducted. population dynamics. Accordingly, future Body weight gains during infections with efforts must investigate the effects of host the highest doses were up to 30% less than age, immunity and genetics, intensity of uninfected sham controls. Pathogenicity exposure, density-dependent mechanisms, of Eimeria innocua may have been grossly and climate and season of infection underappreciated in past literature. on establishment of EL3s, sequential development and survival of L4, L5, and adult stages, and associated egg production. OA38 Equine Cyathostomes II Anthelmintic therapy is widely considered July 10, 2019, 13:30 - 15:30 to be an essential tool for successful Breakout Room 3, Hall of Ideas F&I, Level 4 strongyle control in horses. But currently, the sustainability of this approach is threatened by almost universal resistance among OA38.01 Equine Cyathostomins 1: Relative cyathostomins to benzimidazoles, prevalent Importance and Challenges to Sustainable and expanding resistance to pyrimidines, Control and indications that macrocyclic lactones may be in early decline. Egg counts are the Dr. Craig Reinemeyer1, Dr. Dave Leathwick2, 2 3 current, gold standard measure of success, Dr. Christian Sauermann , Dr. Martin Nielsen but the overall benefits of control in terms 1East Tennessee Clinical Research, Inc., of reducing infection intensity, promoting Rockwood, United States, 2AgResearch, host immunity, and slowing the development Grasslands Research Centre, Palmerston of anthelmintic resistance remain nebulous North, New Zealand, 3M.H. Gluck Equine and perhaps even counter-intuitive. Future Research Center, Dept. of Veterinary Science, cyathostomin control strategies must address University of Kentucky, Lexington, United the dual challenges of significantly decreasing States environmental contamination and minimizing concurrent selection for anthelmintic Cyathostomin nematodes are ubiquitous resistance. parasites of horses and constitute the major target of helminth control for mature, managed equids. Cyathostomins are usually minor pathogens, but large burdens of encysted stages may cause

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 139 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA38.02 Equine Cyathostomins 2. What Does Thus, the model reproduces output typical of Building and Running the Model Tell Us? different climatic regions and suggests that observed patterns of arrested development Dr. Dave Leathwick1, Dr. Christian Sauermann1, may simply reflect the seasonality of free- Dr. Craig Reinemeyer3, Dr. Martin Nielsen4 living stages on pasture as determined by 1Agresearch, Palmerston North, New Zealand, different weather patterns. 2Agresearch, Palmerston North, New Zealand, 3East Tennessee Clinical Research Inc, Rockwood, United States, 4M.H. Gluck Equine OA38.03 Equine Cyathostomins 3. The Research Center, Department of Veterinary Importance of Genetic and Fitness Science, University of Kentucky, Lexington, Parameters When Modelling Anthelmintic United States Resistance

A model for the dynamics of equine Christian Sauermann1, Martin Nielsen2, Dave cyathostomins requires not only an Leathwick1 understanding of the basic life-history, but 1AgResearch, Palmerston North, New also of the factors which drive population Zealand, 2M.H. Gluck Equine Research Center, change. The dynamics of free-living stages is Department of Veterinary Science, University reasonably straight-forward because these of Kentucky, Lexington, United States respond to temperature and moisture similarly to other parasites. Modelling the parasitic In order to model the development of stages was more difficult due to limited data anthelmintic resistance in cyathostomin on what drives worm dynamics. The completed parasites it is necessary to incorporate genetic model includes 1) a declining establishment mechanisms for resistance into the models. of ingested infective stage larvae as horses However, very little is currently known about age, 2) constant development rates for all the the genetics of resistance to anthelmintics in parasitic stages except the encysted early third these parasites. The first use of this model was stage larvae (EL3), for which development to compare the effect of different assumptions rates are variable to reflect the sometimes regarding the inheritance of resistance on extended arrestment of this stage, 3) negative model outputs. Comparisons were made feedback from adult worms to the L4 stage between single and two-gene inheritance, which prevents L4 maturing to adults and where the heterozygote survival was dominant, results in larval mortality. In the absence intermediate or recessive under treatment, of anthelmintic treatments, the life span of and with or without a fitness disadvantage adult worms is approximately 12 months. associated with the resistance mechanism. Anthelmintic treatment, which removes the Resistance developed fastest when the adult worm burden, allows development of heterozygotes survived anthelmintic treatment L4 to replace the removed worms and an (i.e., were dominant) and slowest when they did acceleration in the development of EL3. not (i.e., were recessive). Resistance was slower to develop when inheritance was poly-genic Model performance is strongly influenced by compared to a single gene, and when there was the rate and seasonal pattern of ingestion a fitness cost associated with the resistance of L3 from herbage. While the adult worm mechanism, although the latter variable was the burden remains relatively stable within a least influential. Importantly, while these genetic year the numbers and proportions of larval factors sometimes had a large influence on the stages increase with the numbers of infective rate at which resistant genotypes built up in the larvae ingested. Further, the seasonal rise model populations, their order of ranking was and fall of encysted stages is largely driven always the same when different anthelmintic by the seasonal pattern of infective larvae use strategies were compared. Therefore, on pasture. Because of this, the model while the described model is unlikely to be a reproduces the contrasting seasonal patterns useful at predicting the timing of resistance of mucosal larvae, typical of temperate development, it should be useful for comparing and tropical environments, using only the different treatment and management strategies appropriate seasonality of larvae on pasture. on their propensity to select for resistance.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 140 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA38.04 Equine Cyathostomins 4: The development and these results call for more Impact of Climate and Seasonality on field research to better understand the Anthelmintic Resistance Development dynamics behind anthelmintic resistance in cyathostomin parasites. Martin Nielsen1, Christian Sauermann2, Dave Leathwick2 1University Of Kentucky, Lexington, United OA38.05 Characterization of the States, 2AgResearch, Palmerston North, New Cyathostominae Community within the Zealand Equine Large-Intestine “Nemabiome”: Morphological Identification and Next- Anthelmintic resistance is widespread in Generation Sequencing. equine cyathostomin populations across the world, and the equine industry is forced Jennifer Bellaw1, Dr. Jocelyn Poissant2, Dr. to abandon traditional parasite control John Gilleard3, Dr. Martin Nielsen1 regimens. Current recommendations aim at 1Department of Veterinary Science, University reducing treatment intensity and identifying Of Kentucky, Lexington, United States, high strongylid egg shedders in a targeted 2College of Life and Environmental Sciences, treatment approach. But, virtually nothing University of Exeter, Exeter, United Kingdom, is known about the effectiveness of these 3Department of Comparative Biology and recommendations, nor their applicability to Experimental Medicine, University of Calgary different climatic regions. This study made Faculty of Veterinary Medicine, Calgary, use of a computer model to evaluate the Canada influence of treatment intensity, climate, and seasonality on the development of Cyathostominae are prevalent equine anthelmintic resistance in cyathostomin nematodes, comprising ~40 distinct parasites. All simulations evaluated the use species in horses, nearly all of which are of a single anthelmintic (e.g., ivermectin) identifiable only by examination of adult over the course of 40 model years. The morphology upon equine necropsy. This study made use of weather station data study characterized distinct Cyathostominae representing four different climatic zones: a communities within the equine large cold humid continental climate, a temperate intestine, utilizing and comparing traditional oceanic climate, a cold semi-arid climate, morphologic identification and a recently and a humid subtropical climate. Initially, the developed next-generation sequencing assay impact of time of the year was evaluated by (NGS). Cyathostominae recovered from the simulating a single anthelmintic treatment cecum, ventral colon, and dorsal colon of administered once a year in any of the twelve 26 horses were identified by morphology months. We then evaluated the impact of or NGS. NGS identified 18 species, while treatment intensities varying between 2 morphological identification yielded 15 and 6 treatments per year. And finally, we species and “immature” and “unidentifiable” evaluated treatment schedules consisting categories. Cyathostomum catinatum, of a combination of strategic treatments Cylicocyclus nassatus, and Cylicostephanus administered to all horses and additional longibursatus together comprised more than selective treatments. Month of treatment had 50% of the entire Cyathostominae community. a large effect on resistance development in Immature stages contributed ~11% to the total colder climates, but little or no impact in the morphologically identified population. Most subtropical climate. Resistance development species exhibited spatial niche preferences was affected by treatment intensity, but was with ~80% or more specimens recovered also strongly affected by climate. Selective from a single gut compartment. Species therapy delayed resistance development in diversity varied significantly between gut all modelled scenarios, but, again, this effect compartments; the cecum was dominated was climate dependent with the largest delays by Coronocyclus coronatus, ventral colon observed in the colder climates. This study is by Cya. catinatum, and the dorsal colon the first to demonstrate an impact of climate by Cys. longibursatus. Diagnostic statistics and seasonality on anthelmintic resistance and method agreement were explored for

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 141 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

11 species. Statistically significant Cohen’s treated or 2) horses were treated if they Kappa values (p≤0.05) were interpreted as exceeded thresholds between 100 and 600 fair (0.21-0.40) for two, moderate (0.41-0.60) strongyle eggs per gram. We imported for six, and substantial (0.61-0.80) agreement weather data representing four different for three species. For seven species, climatic zones: a cold humid continental McNemar X² p-values (p>0.05) showed no climate, a temperate oceanic climate, a cold test disagreement. However, for the three semi-arid climate, and a humid subtropical most abundant species, Cya. catinatum, Cor. climate. Additionally, we evaluated equine nassatus, and Cys. longibursatus, McNemar’s herds with three different age structures; 1) all test showed significant disagreement yearlings, 2) all mature horses 10-20 years old, (p≤0.05). Sensitivity and specificity were and 3) a mixed age structure of 1-20 years of calculated for NGS against gold standard age. Results indicated a clear effect of age morphologic identification. Sensitivity and structure, with resistance developing quickest specificity were variable across species, both in the yearling group and slowest among ranging from ~0.43-0.95% with means of the mature horses. Resistance development 77%. Positive and negative likelihood ratios was affected by treatment intensity and ranged from 1.7-11 and 0.07-0.60, respectively. selective therapy generally delayed resistance. Poor test agreement, low specificity, and However, this effect was more pronounced high negative likelihood ratios are influenced in the mature horses compared to the two by a high rate of “false positives,” resulting other age structures. Finally, the effects of from NGS identifying immature or otherwise selective therapy on resistance development morphologically unidentifiable specimens. were also climate dependent with resistance Further development and validation of NGS being delayed least in the subtropical climate with previously morphologically identified and most in the other climates. These results specimens is warranted. suggest that defining an optimal selective treatment strategy is more complicated than simply applying an arbitrary threshold OA38.06 Equine Cyathostomins 5: Benefits across all horses and environments, and of Selective Anthelmintic Therapy are Age more work is needed to provide meaningful and Climate Dependent recommendations.

Martin Nielsen1, Christian Sauermann2, Dave Leathwick2 OA38.07 Outbreak of Acute Larval 1University Of Kentucky, Lexington, United Cyathostominosis After Anthelmintic States, 2AgResearch, Palmerston North, New Treatment: Investigation of the Clinico- Zealand Pathological Parameters and the Faecal Microbiome Changes in Twelve Horses Selective anthelmintic therapy has been recommended as a sustainable strategy for Ms. Nicola Walshe1, Professor Grace cyathostomin control in horse populations Mulcahy2,4, Dr. Fiona Crispie3, Dr. Paul Cotter3, for several decades. By determining Dr. Raul Cabrera Rubio3, Associate Professor strongyle fecal egg counts (FEC) for all Vivienne Duggan1 horses, and only treating those exceeding a 1School of Veterinary Medicine, University predetermined FEC threshold, the aim is to College Dublin, Ireland, 2School of achieve a reduction of overall egg shedding, Veterinary Medicine, University College while leaving a proportion of the herd Dublin, Ireland, 3Teagasc Moorepark/ untreated. This is based on the overdispersed Alimentary Pharmacobiotic Centre, University distributions of fecal egg counts across herds College Cork, Ireland, 4Conway Institute and the consistent shedding levels maintained of Biomolecular and Biomedical Research, by individual horses across time. We used Belfied, Ireland the cyathostomin model to evaluate the influence of selective treatment strategies Cyathostomins are the most common and with between 1 and 4 yearly treatment pathogenic endoparasites in horses. Within occasions, where either 1) all horses were the cyathostomin life cycle, hypobiosis of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 142 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

encysted larvae in the large intestinal mucosa This study provides further evidence of an can occur, allowing for accumulation of interplay between cyathostomin infection, large burdens. One clinical presentation is systemic inflammation and gut homeostasis in a severe local and systemic inflammatory the horse. syndrome – acute larval cyathostominosis (ALC). The risk factors and pathophysiology of ALC are incompletely understood, but OA38.08 Cyathostomin Infection Influencing it can be precipitated by administration of Growth Rate and Performance of anthelmintics. Thoroughbred

We have previously documented systemic and Dr. Marcelo Molento1, Dr. Luciana Dias local inflammation and decreased diversity de Castro1, Carolina Abrahao1, Dr. Paulo of the faecal microbiota, in clinically normal Guimaraes1, Joaquim Antunes2 horses hosting encysted larval cyathostomins. 1UFPR, 157, Brazil, 2Private Practitioner, Sao We hypothesised that similar, but greater, Jose dos Pinhais, Brazil changes in these parameters would be seen in horses which develop ALC. Here, we Cyathostomin infection are ascribed to document the clinico-pathological and faecal cause impairment to body development and microbiota changes during an ALC outbreak performance of foals, but the actual effect of in Ireland. the disease remains unclear. The aim of this study was to determine the growth rate and The outbreak of ALC in a herd of twenty- racing performance of 31 naturally infected three horses began in mid-November Thoroughbred horses, and to correlate with approximately three weeks after treatment gender, month of birth, age, number of with ivermectin/praziquantel. Clinical signs offspring per mare, fecal egg count (FEC) included sudden weight loss, diarrhoea, dull and packed cell volume (PCV), from birth to demeanour, pyrexia, colic and vasculitis. 3.8 years of age. Contents: Body weight (BW) Clinico-pathological changes included and withers height (WH) were conducted neutrophilic and lymphocytic leucocytosis, monthly, PCV was measured from 12-21 hyper-fibrinogenemia, hyper-globuinemia, months of age, and FEC from 9-21 months hypo-albuminemia and an albumin:globulin of age. Performance was determined by reversal. the Spearman’s correlation, considering the number of races between Dec/2015 and Faecal bacterial microbiota were Mar/2017, and previous FEC. Correlation characterised by 16sRNA sequencing in between all variables was analyzed using longitudinal samples taken throughout the Spearman’s rank correlation. All animals outbreak, in both affected and clinically were positive for cyathostomins, and FEC normal horses. ranged from 389 – 1.635. Average BW at birth was 56.55 kg for males and 51.9 kg for Affected horses were treated with moxidectin females, and at 18 months’ males and females and corticosteroids. Supportive therapy was weighed an average of 435.89 and 441.65 administered according to clinical status. kg, respectively. Average WH at 18 months Broad spectrum antimicrobial therapy was the mean height was 154.78 and 153.5 cm, administered to horses with persistent respectively. Cyathostomin infection did not pyrexia. appear to have affected the growth rate of the animals, as the correlations between FEC, Of the twelve clinically-affected horses, BW and WH were not significant. The number ten recovered and two were euthanised. of offspring per mare influenced the weight Postmortem findings showed a marked of the animals at birth (p = 0.03) and at 6 ulcerative typhlocolitis consistent with the months of age (p = 0.026). The racing history mass emergence of cyathostomin larvae (n=30) data revealed that animals with high and evidence of opportunistic bacterial FEC showed poorer (p<0.002) performance pathogenesis. when compared to those with low FEC with a coefficient of -0.580. Conclusions: The current

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 143 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

study provides strong evidence regarding relation to a susceptible reference strain. the expected growth rate of Thoroughbred Fisher’s test was used to determine the foals in Brazil, and the racing performance correlation of resistance among the drugs. associated FEC, which may have important There was correlation in the resistance breeding implications. between ivermectin and doramectin (p=0.04) in 19 samples, and there was 2.3-fold (95%CI: 1 to 5.4-fold) more chances of a tick population OA39 Insecticide and Acaricide to be resistant to doramectin if the same Resistance in Ruminants is resistant to ivermectin. There was no correlation of resistance between ivermectin July 10, 2019, 13:30 - 15:30 and moxidectin (p=0.24) nor between Breakout Room 4, Hall of Ideas G&J, Level 4 doramectin and moxidectin (p=0.67). This is the first study to demonstrate side-resistance among avermectins in R. microplus. The OA39.01 Macrocyclic Lactones Side- results obtained here have an impact on cattle Resistance in Isolates of Rhipicephalus tick control strategies, since side-resistance Microplus From Rio Grande Do Sul, Brazil between avermectins limits the availability of

1 1 drugs to be used. Contrarily, moxidectin can Marcelo Becker , Paola Antunes , Manoel still be rationally used as a control measure. Poitevin3, Dr. Jose Reck1, Prof. Tiago Gallina3, 1,2 Further studies should be performed to Dr. Guilherme Klafke elucidate the mechanisms of resistance and 1Instituto de Pesquisas Veterinarias Desiderio side-resistance among different macrocyclic Finamor, Secretaria da Agricultura do Rio lactones. Grande do Sul, Eldorado Do Sul, Brazil, 2USDA-ARS Catlle Fever Tick Research Laboratory, Edinburg, United States, OA39.02 Detection of Resistance to 3Universidade Federal do Pampa (Unipampa), Chemical Acaricides in Rhipicephalus Uruguaiana, Brazil (Boophilus) Microplus Ticks From Selected Cattle Farms in Luzon, Philippines The Rio Grande do Sul (RS) state plays a major role in Brazilian cattle industry. The Dr. Remil Galay1, Dr. Sherwin Alota1, Dr. Tisha parasitism by the cattle tick, Rhipicephalus Rogelle Edquiban1, Dr. Jayvee Evangelista1, microplus, causes great economic losses Mr. Fred Gio Rean Valera1, Dr. Kristina Andrea to the producers in the area. In order to Sandalo1, Dr. John Michael Bernardo1, Dr. minimize damage and optimize the chemical Tetsuya Tanaka2 control, it is fundamental to monitor the 1College of Veterinary Medicine, University acaricide resistance status. The resistance of the Philippines Los Banos, Los Banos, of R. microplus to macrocyclic lactones Philippines, 2Joint Faculty of Veterinary has been detected in RS for 17 years. Side- Medicine, Kagoshima University, Kagoshima, resistance among the drugs of this class Japan is an important condition, considering its implications in resistance management and Tick infestation and tick-borne diseases cattle tick control strategies. The aim of cause great economic losses in the this study was to evaluate the occurrence cattle industry worldwide. The cattle tick of side-resistance among avermectins Rhipicephalus (Boophilus) microplus is (ivermectin and doramectin) and milbemycins one of the most widely distributed ticks, (moxidectin) in R. microplus. Engorged female especially in tropical countries including ticks were sampled in 26 premises and their the Philippines. Tick infestation remains to larvae were used in larval immersion tests be a burden to many cattle raisers, leading with ivermectin, doramectin and moxidectin. to health problems, decreased productivity Mortality data was submitted to probit and economic losses. Tick control is mainly analysis to calculate the lethal concentrations through application of chemical acaricides (LC50) and 95% confidence intervals (CI). such as amitraz and synthetic pyrethroids. Resistance ratios (RR) were calculated in Ivermectin is also widely used to control

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 144 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

gastrointestinal nematodes, and partly for the world where livestock is a principal tick control. Acaricide resistance has been activity with great veterinary and economic reported in several countries, but there are no importance. One of the main tick control published reports in the Philippines. Hence, measures is the use of acaricides and this study aimed to determine whether there macrocyclic lactones (ML). In Mexico no is resistance already to various chemical information on the distribution of level of acaricides that are being used. Engorged or acaricide and ML resistance in R. microplus nearly engorged R. (B.) microplus female field populations is available. For this study ticks were collected from selected cattle field tick populations were collected from farms in provinces in northern and southern 42 farms in 12 different states of Mexico and Luzon island of the Philippines, which were level of resistance to different acaricides and allowed to lay eggs in the laboratory to macrocyclic lactone determined. The dose- produce larvae. The resulting larvae were response bioassays were carried out using exposed to various concentrations of amitraz, the modified larval packet (coumaphos and cypermethrin and ivermectin through the flumethrin) and larval immersion test (amitraz larval packet test (LPT). The concentrations and ivermectin) against R. microplus. Mortality included discriminating dose (DD), half of data were subjected to probit analysis to DD, and double of DD. Larval mortality was calculate lethal concentrations at 50%. A determined after 24 hours. LPT for amitraz logistic regression model was used to evaluate showed more than 90% larval mortality the relation between resistance and possible for all farms tested. Meanwhile, LPT for associated factors. Phenotype was defined as cypermethrin showed more than 90% larval susceptible, low resistance or high resistant. mortality in DD and double DD in all except The overall prevalence of cattle farms for three farms tested, wherein the mortality with R. microplus resistant to coumaphos, for DD was below 80%, suggesting presence amitraz, flumethrin and ivermectin were of resistance to cypermethrin. In case of 19.0%, 52.4%, 45.2% and 85.7%, respectively. ivermectin, the mortality of tick larvae from For coumaphos, 80.9%, 14.3%, and 4.8% five farms was lower than 80% even in double were susceptible, low resistance and high of DD, suggesting resistance. Additional farms resistance, respectively; for amitraz, 47.6%, from other provinces are currently being 26.2%, and 26.2%, respectively; for flumethrin, tested. This study provides the first scientific 54.8%, 23.8%, and 21.4%, respectively; and evidence of R. (B.) microplus resistance for ivermectin, 14.3%, 61.9%, and 23.8%, to cypermethrin and ivermectin in the respectively. We identified that cattle farms Philippines. without acaricide rotation program (OR: 9.16, CI95%: 1.70-48.40, P: 0.009) had higher probability of developing R. microplus OA39.03 Monitoring the Resistance of resistant to amitraz. It is concluded that Rhipicephalus Microplus to Conventional amitraz, flumethrin and ivermectin resistance Acaricides (Coumaphos, Amitraz and in R. microplus is common, but mainly at low Flumethrin) and Ivermectin on Cattle Farms level in cattle farms of Mexico. Besides the in Mexico intensive use of coumaphos for many years in the control R. microplus in cattle from Mexico, Professor Roger Ivan Rodriguez-Vivas1, Dr. this acaricide can still be used as a tool to Eduardo Eduardo Ramirez Espana2, Dr. Ivan control the cattle tick. Lozano Blanco2, Dr. Melina M. Ojeda-Chi1, QFB Iris Trinidad-Martinez1, Dr. Juan Agustin Torres Research founded by Bayer Animal Health, Islas2, Dr. Chandra Bhushan3 Germany. 1Universidad Autonoma De Yucatan, Merida, Mexico, 2Bayer Animal Health, Mexico, México city, Mexico, 3Bayer Animal Health, Germany, Leverkusen, Germany

Rhipicephalus microplus is widely distributed in tropical and subtropical regions of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 145 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA39.04 Mapping Acaricide Resistance also resistant to alpha-cypermethrin. Only of the Three Major Cattle Tick Species LD50 for A. variegatum could be determined Amblyomma Variegatum, Rhipicephalus because of the absence of a susceptible Appendiculatus and Rhipicephalus Microplus strain. in East and West Africa by Using the Larval Packet Test (LPT) The degree of resistance and use of the different acaricides in each country is Dr. Maxime Madder1, Dr. Theo Schetters1, Dr. discussed. Josephus Fourie2, Dr. Frans Jongejan3 1Clinglobal, Black River, Mauritius, 2Clinvet, Bloemfontein, South Africa, 3FAO Reference OA39.05 Efficacy of a New 2.5% Fluazuron, Centre for Ticks and Tick-borne Diseases, 7% Chlorpyrifos, 6% Cypermethrin, University of Utrecht, Utrecht, The 6% Piperonyl Butoxide-based Pour- Netherlands On Formulation Against Rhipicephalus (Boophilus) Microplus in a Cattle Population Acaricide resistance is a major constraint under Field Conditions in Brazil in the control of ticks on production and companion animals all over the world. A Dr. Fabio Barbour Scott2, Dr. Gabriela Ferreira variety of reasons contribute the development de Oliveira2, Dr. Barbara Rauta de Avelar2, of resistance, among which long-time use, Márcia Cristina Alves Teixeira3, Dr. Marcio under- or overdosing or inappropriate use. Dentello Lustoza3, Dr. José Manuel Moreno In this study the degree of acaricide Montroy4, Dr Emmanuel Briant1, Dr. Christelle resistance of three economically most Navarro1, Dr. Vanessa Chala1 important cattle tick species (Amblyomma 1Virbac, Carros, France, 2Universidade Federal variegatum, Rhipicephalus appendiculatus Rural do Rio de Janeiro, Seropédica, Brasil, and R. microplus) collected in resource- 3Virbac do Brasil Industria e Comercio LTDA, poor farmer communities in East and West São Paulo, Brasil, 4Laboratorios Virbac Mexico, Africa were determined. In seven countries Guadalajara, Mexico (Benin, Ethiopia, Ghana, Nigeria, Tanzania and Uganda) and at two localities in each country In Brazil Rhipicephalus microplus in cattle known for its high livestock density and causes serious economic losses. The objective occurrence of ticks and tick-borne diseases, of this study was to test the efficacy of engorged females belonging to these species, a single pour-on treatment with a new if present, were collected. Their subsequent formulation 2.5% Fluazuron, 7% Chlorpyrifos, offspring was used in larval packet tests to 6% Cypermethrin, 6% Piperonyl Butoxide at determine LD50 and degree of resistance, 1 mL/10kg/Bw on the elimination of a natural when compared to a susceptible tick strain. tick infestation in a Brazilian cattle population. For A. variegatum, six different tick stocks were analysed (one from each country); for For this study, 22 male or female cross- R. appendiculatus five (three from Uganda breed animals Bos indicus x Bos taurus, 24 and two from Tanzania) and for R. microplus to 36 months old, 190-340 kg live weight 10 different stocks (two from each country). were selected from the experimental herd To determine the degree of resistance, a of Universidade Federal Rural do Rio de susceptible strain of R. appendiculatus and Janeiro. Animals were kept on pastures, R. microplus, both from South Africa, were without parasiticide treatment and were included in the tests. naturally infested by Rhipicephalus microplus. On D0, these animals were randomized to Five different acaricides were tested: the control group (C, n=11) or the treatment chlorfenvinphos, alpha-cypermethrin, group (T, n=11) with consideration to the fipronil, ivermectin and amitraz. The results average number of engorged females ticks of the LPT indicate an intermediate and (ø ≥ 4.5 mm) collected on D-3, D-2, D-1. On high level of resistance for R. microplus and D0, 1 mL/10 kg/Bw of this new formulation R. appendiculatus against chlorfenvinphos containing 2.5% Fluazuron, 7% Chlorpyrifos, and amitraz. Rhipicephalus microplus was 6% Cypermethrin, 6% Piperonyl Butoxide

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 146 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

(Virbac) was poured on the back of group T in mite counts and the cure rate after the first animals, from the base of their horns to the treatment round and the number of treatment base of their tails. Once a week, from day D+7 rounds needed to cure all animals. In vitro until D+70, engorged females ticks (Ø ≥ 4.5 knock-down and mortality was evaluated in mm) were counted on animals. a contact assay based on Brimer et al., 1995 (Vet Parasitol 59, 249-255). Between D-3, D-2, D-1 the average tick counts in geometric means were 70.6 and 69.8 All farms needed ≥ 2 treatment rounds (2- in groups T and C, respectively. In treated 8) to obtain full efficacy. Cure rates varied animals, the tick population decreased from 0%-80%. Only three farms had a mite significantly (p<0.05) in comparison with the count reduction of >90%, two of which had a control group for all time points from days lower limit of the confidence interval of <90%. D+7 until D+70, and in geometric means, by All other farms had a mite count reduction 97.4, 98.6, 96.7, 98.7, 97.5, 96.4, 98.6, 97.15, <90% (-411%-81%). LD50 values in vitro varied 92.2, and 81.1% respectively. from 2,951-36,867 g/mL and 0.3-58.3 g/mL at 24h and 120h, respectively. No significant In this study, this new pour-on formulation correlation was found between in vitro LD50 was found to be effective against values and any of the parameters for in vivo Rhipicephalus Microplus with a reduction of efficacy. tick counts by more than 95 % until 56 days In conclusion, unambiguous treatment failure after the treatment. was detected on 13/16 beef farms, confirming the presence of ML resistance in Belgian Blue beef farms. Tentative in vitro parameters OA39.06 In Vivo and In Vitro Parameters for could not detect ML resistance. The potential Acaricide Efficacy of Macrocyclic Lactones use of different in vitro and in vivo parameters Against Psoroptes Ovis in Cattle to evaluate acaricide efficacy and to detect acaricide resistance will be discussed. Wouter Van Mol1, Nathalie De Wilde1, Stijn Casaert1, Zhen Zhen Chen1, Marieke Vanhecke1, Prof. Dr. Luc Duchateau2, Prof. Dr. Edwin OA39.07 Acaricidal Activity of Nanoscale Claerebout1 ZnO Encapsulated Piperine Formulation 1Department of Virology, Parasitology and Against Rhipicephalus Microplus Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium, Dr. Vukka Chengalva Rayulu1, Dr. Kancharana 2Department of Nutrition, Genetics and Snigdha1 Ethology, Faculty of Veterinary Medicine, 1Sri Venkateswara University, Tirupati, India Ghent University, Merelbeke, Belgium A study was undertaken with an aim to Psoroptic mange is an important disease synthesize and evaluate the acaricidal in beef cattle, and Belgian Blue cattle are activity of the nanoscale zinc oxide piperine particularly susceptible. Treatment failure of formulation (NZPF) against Rhipicephalus macrocyclic lactones (ML) against Psoroptes microplus ticks infesting cattle. NZPF was ovis has been reported, but clear evidence prepared by employing encapsulation for resistance of P. ovis against ML in cattle is technique using 0.1% zinc oxide nanoparticles lacking. This study was conducted to investigate (ZnONPs) and 2% piperine solution. The ML efficacy in 16 beef farms in Belgium and The synthesised NZPF was characterized by Netherlands in vivo and in vitro. subjecting UV–Vis spectroscopy, Fourier Transformed Infrared analysis, X-ray On each farm a group of animals (n= 7-14) Diffraction, Dynamic Light Scattering, with clinical psoroptic mange was treated Scanning Electron Microscopy and Energy with two subcutaneous injections of a short- Dispersion Spectroscopy analysis. Acaricidal acting ML with 7-10 days interval (n=15) or a activity of deltamethrin, piperine, ZnONPs single injection with a long-acting ML (n=1). In and NZPF on Rhipicephalus microplus was vivo efficacy was assessed by the reduction evaluated by two bioassays viz., Larval

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 147 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Packet Test (LPT) and Adult Immersion Test The current work assessed the relationship (AIT). LPT with a discriminating dose of between pharmacokinetic behavior and deltamethrin (75ppm) showed a mortality clinical efficacy of ivermectin (IVM) or of 59% of R. microplus larvae. Mortality of R. doramectin (DRM) against natural Psoroptes microplus larvae was 100% at concentration ovis var. bovis infection in cattle. The study of 9 ppm, 8 ppm and 7 ppm with piperine, involved two trials (I and II) carried out on ZnONPs and NZPF, respectively. AIT with a different beef cattle production systems, a discriminating dose of deltamethrin (75ppm) feedlot (Trial I) and a grazing (Trial II) system. against adult R. microplus showed a mortality In Trial I, 40 mange-infected steers were of 40%, oviposition inhibition of 78.309% and allocated into 4 groups (n=10) and treated the lowest egg mass weight with 17.8±1.31 mg. with a single (day 0) or repeated (days 0 and Mortality rate and oviposition inhibition of R. 7) subcutaneous injection of two different microplus were 100% whereas egg mass and formulations of IVM (1%) at 0.2 mg/kg. In reproductive index were completely nil with Trial II, 20 grazing calves with active mange both piperine and ZnONPs at a concentration infection were allocated into 2 groups (n=10) of 20 ppm and NZPF at a concentration of and treated with a single subcutaneous 15 ppm. NZPF showed a potent ovulation injection of either IVM (1%) or DRM (1%) at inhibitory activity with significantly (P<0.05) 0.2 mg/kg. Blood and skin samples were lower IC50 and IC99 values compared to collected from 8 animals of each group to ZnONPs and piperine. measure IVM/DRM concentrations by HPLC. Skin scraping samples were collected from Both LPT and AIT results indicated the each animal and mites were counted. In Trial I, development of resistance in R. microplus the repeated administration of IVM increased ticks against deltamethrin. NZPF, ZnONPs the systemic availability and skin drug and piperine were found to have significantly exposure compared to the single treatment (P<0.05) higher acaricidal activity. However, (p<0.05). However, both formulations failed NZPF had high acaricidal efficacy at lower to achieve a clinical mange cure at either concentrations than pure phytochemical single or repeated treatment. Efficacy of IVM piperine, ZnONPs and deltamethrin. NZPF was 10% (single dose) and 50% (repeated could be potential alternative to routine treatment) at day 14 post-treatment. The non- chemical acaricides for control of tick cured animals remained with active mange 28 infestation of cattle in the wake of the days post-treatment. No differences (p>0.05) development of acaricidal resistance, residual in the P. ovis scores density were observed effect and environmental pollution. after single or repeated treatments. In Trial II, there was also a positive correlation between IVM or DRM concentrations in plasma and OA39.08 Failure of Macrocyclic Lactones to skin samples. Although IVM and DRM failed Control Psoroptic Mange Infection in Feedlot to obtain a complete parasitological cure, the and Grazing Beef Cattle efficacy of DRM was higher (80%) than those obtained by IVM (10%)(p<0.05).Additional Dr. Candela Canton1, Dr. Cesar Fiel2, Dr. Pedro studies are needed to confirm the presence Steffan2, Veterinarian Sebastian Muchiut2, of P. ovis populations resistant to macrocyclic Veterinarian Maria Paula Domínguez1, Dr. lactones, and to enhance the control of Guillermo Virkel1, Veterinarian Laureano psoroptic mange in cattle. Schofs1, Dr. Carlos Lanusse1, Dr. Luis Ignacio Alvarez1, Dr. Adrian Lifschitz1 1Laboratorio de Farmacología, Centro de Investigación Veterinaria de Tandil (CIVETAN), UNCPBA-CONICET-CICPBA, Tandil, Argentina, 2Laboratorio de Parasitología y Enfermedades Parasitarias, Centro de Investigación Veterinaria de Tandil (CIVETAN), UNCPBA- CONICET-CICPBA, Tandil, Argentina

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 148 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA40 Zoonoses infections. Pk, Pcy and Pin were detected either singly or in combination in all the July 10, 2019, 13:30 - 15:30 malaria-positive macaques. This study Breakout Room 5, Meeting Rooms KLOP, Level 4 indicates that the simian malaria parasites that can infect humans, namely P. knowlesi, P. cynomolgi and P. inui, are found in macaques OA40.01 Zoonotic Malaria Parasites Among throughout Sarawak, Malaysian Borneo. Non-Human Primates in Sarawak, Malaysian Travelers to forested areas in Sarawak and Borneo similar areas in Southeast Asia should be made aware of the potential risk of acquiring 1 Ms. Khamisah Abdul Kadir , Dr. Thamayanthi zoonotic malaria. Nada Raja1, Dr. Paul Cliff Simon Divis1, Mdm Dayang Shuaisah Ahmad Mohamad1, Mr 1 Ahmad Matusop , Professor Mohd Tajuddin OA40.02 Neuroinflammation and 3 1 Abdullah , Professor Balbir Singh Neurodegeneration during Toxocara Canis 1 Malaria Research Centre, Universiti Malaysia and T. Cati-Induced Neurotoxocarosis in Mice Sarawak, Kota Samarahan, Malaysia, 2Sarawak Health Department, Kuching, Malaysia, Andrea Springer1, Patrick Waindok1, Lea 3 Universiti Malaysia Terengganu, Kuala Nerus, Heuer1,2, Elisabeth Janecek-Erfurth1, Fred Malaysia Lühder3, Andreas Beineke4, Christina Strube1 1Institute for Parasitology, Centre for Human infections with Plasmodium knowlesi, Infection Medicine, University of Veterinary a malaria parasite of long-tailed and pig- Medicine Hannover, Hannover, Germany, tailed macaques (Macaca fascicularis and 2Bayer Animal Health GmbH, Leverkusen, M. nemestrina respectively), were thought Germany, 3Department of Neuroimmunology, to be extremely rare until a large focus Institute for Multiple Sclerosis Research, The of human infections were reported in the Hertie Foundation, Göttingen, Germany, Kapit division of Sarawak, Malaysian Borneo 4Department of Pathology, University of in 2004. Macaques are natural hosts of P. Veterinary Medicine Hannover, Hannover, knowlesi (Pk), P. cynomolgi (Pcy), P. inui Germany (Pin), P. coatneyi (Pco), P. fieldi (Pfi) and P. simiovale. In Sarawak, only macaques in the Migrating larvae of Toxocara spp. may cause Kapit division have been examined so far and disease in humans, including neurotoxocarosis found to harbor these malaria parasites. The (NT). Previous studies on T. canis- and T. objective of this study was to determine the cati-induced NT in the model organism prevalence and distribution of simian malaria “mouse” have indicated distinct differences parasites among non-human primates in between both Toxocara species regarding other divisions in Sarawak, Malaysian Borneo. larval migration patterns and clinical From 7 out of the 9 administrative divisions in symptoms. The aim of the present study was Sarawak, blood samples were obtained from to investigate patterns of neuroinflammation 84 non-human primates: Macaca fascicularis and neurodegeneration during T. canis- and (n=45), M. nemestrina (n=28), Presbytis T. cati-induced NT in the mouse model. cristata (n=2), Nasalis larvatus (n=2), Macaca Histopathology was carried out on cerebra arctoides (n=3), Hylobates muelleri (n=4). and cerebella of Toxocara-infected and They were screened with nested PCR assays uninfected C57Bl/6 mice at 7, 14, 28, 42, using primers specific for Pk, Pcy, Pin, Pco and 70 and 98 days post infection (dpi). In Pfi. Only macaques were found to be infected addition to haematoxylin-eosin and luxol- with malaria parasites and they originated fast-blue staining, immunohistochemistry from all 7 Sarawak divisions sampled. A was employed to visualize microglia and total of 31 macaques (20 M. fascicularis and beta-amyloid precursor protein (beta- 11 M. nemestrina) were malaria-positive. All APP) as an indicator of axonal damage. 5 Plasmodium species were detected, with Furthermore, a comprehensive multiplex macaques harbouring either single (n=12), bead array assay profiling was conducted double (n=8), triple (n=7) or quadruple (n=4) to elucidate patterns of immunomodulatory

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 149 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

cytokines and chemokines in affected spleen of some animals. We investigated the mouse brains. Haemorrhages, eosinophilic cause of lesions in sites other than the muscle vasculitis and activated microglia were or brain in a total of 157 pigs from T. solium detected in both infection groups starting endemic regions of Uganda and Nepal which 7 dpi, followed by eosinophilic meningitis were subjected to extensive investigations at 14 dpi. Neurodegenerative processes necropsy. Lesions which had the potential to (demyelination, beta-APP accumulation, gitter be caused by T. solium were characterised by cells) occurred earlier during T. canis than macroscopic and microscopic examination, T. cati infection, and affected significantly histology as well as DNA characterisation more T. canis- than T. cati-infected mice, in PCR-RFLP and sequencing. Lesions especially regarding the cerebrum. In both were confirmed as being caused by Taenia infection groups, a continuous decrease of hydatigena (both viable and non-viable), certain pro-inflammatory cytokines, including by T. asiatica and Echinococcus granulosus TNF- , IFN- -and IL-12, was detected in the (in Nepal) and nematode infections. No cerebrum over the course of infection, while cysticercus or equivocal lesion was found differencesα γ were detected regarding Th2- to have been due to T. solium in any tissue cytokines. T. canis infection was characterised location other than muscle and brain. It by significantly elevated IL-4 and IL-5 levels in is recommended that future evaluations the cerebra in the acute and subacute phase of porcine cysticercosis in aberrant tissue of the disease, which were not detected in T. locations involving DNA analyses take cati-infected mice. Further differences were appropriate care to avoid the possibility of observed, for example, regarding eotaxin. contamination of tissue specimens with DNA Earlier and more severe neurodegeneration from a different tissue location or a different during T. canis- than T. cati-induced NT animal, and the use of appropriate control may explain the differences in behavioural samples in order to confirm the absence of alterations observed in previous studies. cross sample contamination. In addition to larval migration preferences, immune regulatory mechanisms may contribute to these patterns. OA40.04 Prevalence of Ticks Parasitizing Human Beings in the Nilgiris Hills of Southern India OA40.03 Accurate Diagnosis of Lesions Suspected of Being Caused by Taenia Dr. Soundararajan Chinnaiyan1, Dr. Prabhu Solium in Body Organs of Pigs with Naturally Kandasamy Perumal2, Dr. Nagarajan Acquired Porcine Cysticercosis Kumaresan3, Dr. Arul Prakash Mani4 1Tamilnadu Veterinary and Animal Sciences Marshall Lightowlers1 University, India, 2Tamil Nadu Animal 1Faculty of Veterinary and Agricultural Husbandry Department, India, 3Tamilnadu Sciences, Australia Veterinary and Animal Sciences University, India, 4Tamilnadu Veterinary and Animal The definitive method for diagnosis of Sciences University, India porcine cysticercosis is detection of cysticerci at necropsy. Cysts are typically located Ticks act as a vector to transmit varieties in the striated muscle and brain. Until of pathogenic organisms to human beings recently Taenia solium cysticerci have not viz., Kyasanur forest disease ad congo been definitively identified in other tissue haemorrhagic fever. A study was conducted locations, despite several comprehensive to know the prevalence of tick species investigations having been undertaken which on 92 human beings (56 male and 36 included investigation of body organs other females) working in the tea estate, fire wood than muscle and brain. Recently a study collectors, reserved forest, coffee /pepper conducted in Zambia reported 27% infection pickers, persons residing in home and also with T. solium in the liver of pigs with naturally on the premises at Pallipadi, Irumbupalam, acquired porcine cysticercosis, as well as Karalikandi, Devala, Pandiyar Tan tea 2B, some T. solium infection in the lungs and Sussex (Pitheri), Genepool (Nadukani),

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 150 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Erumadu and Mundakunnu of Gudalur, the of the Environment, Japan, and municipal Nilgiris district of Tamil Nadu, India from governments, including Kanagawa Prefecture, January 2018 to February 2019. Overall are implementing control measures to reduce prevalence of tick infestation on human their numbers. Enterocytozoon bieneusi is a beings was 53.26%. The collected ticks were common microsporidian species, frequently identified as larval and nymphal stages of reported from immunocompromised humans Haemaphysalis spinigera, H. megalaimae, and from a wide range of domestic and wild Haemophysalis Spp., Amblyomma Spp,, animals. More than 240 genotypes have Anomalohimalaya Spp. and nymphal stage been identified in both humans and animals of Amblyomma varanense and Hyalomma worldwide; however, information regarding isaaci by stereo zoom and scanning electron E. bieneusi in mammals in Japan is scarce. microscope (SEM). Ticks collected from the We aimed to determine the occurrence and premises by tick drag and tick flag methods genotype of E. bieneusi in Pallas’s squirrel were identified as larval and nymphal stages from Kanagawa, Japan for the first time. of H. spinigera. Persons infected with ticks Genomic DNA was extracted from a total showed fever and vomition until removal of of 168 feces samples from Pallas’s squirrels ticks and reluctant to work, severe itching, that were captured in Hayama City between tiredness, inappetance and the skin lesions January and June 2018. Nested PCR was persist from a week to more than two years. performed targeting the ribosomal internal Erupted, thickened brownish gray scar (2 transcribed spacer (ITS) region. The overall to 5 mm) on chest region and wide-spread prevalence was 16.7 % (28/168), and sequence multifocal lesions on the back from the neck analysis showed 100 % homology to genotype to hip region, abdomen, shoulder and hands SCC-2 for all 28 samples. This genotype were seen. The affected persons working in was recently reported from pet chipmunks the reserve forest region revealed multifocal (Eutamias sibiricus asiaticus) in Sichuan erythematous nodular eruptions in the leg and Province, China, which phylogenetically thigh region. The prevalence ticks found to clustered into a novel group (Group 10). It is be more on the persons working in tea estate not known whether this genotype poses a (42.86%) followed by persons residing in risk of zoonotic transmission and can cause home (20.41%), fire wood collectors (18.37%), human microsporidiosis. As Pallas’s squirrels reserved forest (18.37%) and no ticks were in Kanagawa Prefecture live in close proximity observed on coffee /pepper pickers. to humans, it is important to be aware of the Key words: Ticks – human – prevalence – potential risk from a public health perspective. lesions – southern India Continuing studies on the distribution of E. bieneusi genotypes in the squirrel population in Japan are necessary to further assess the OA40.05 Molecular Characterization of effect of the pathogen on both wildlife and Enterocytozoon Bieneusi in Pallas’s Squirrels human populations. (Callosciurus Erythraeus) from Kanagawa Prefecture, Japan OA40.06 Improved Diagnostics for Aya Masuda1, Yui Fuwa1, Fumiaki Yamasaki2, Echinococcus Multilocularis in Wild and Jun Matsumoto1 Domestic Canid Species. 1Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, Temitope Kolapo1, Dr. Caroline Frey2, Joy Wu1, Japan, 2School of Veterinary Medicine, Nippon Mila Bassil1, Brent Wagner1, Dr. Jason Acker3, Veterinary and Life Science University, Japan Dr. Emily Jenkins1 1University of Saskatchewan, WCVM, Canada, Pallas’s squirrel (Callosciurus erythraeus) 2Canadian Food Inspection Agency, Canada, was introduced from Taiwan to Japan in the 3Laboratory Medicine and Pathology, Canada 1930s and eventually established itself in several areas across the country. The species In recent times, there has been an increasing is designated as a regulated organism under detection of alveolar echinococcosis (AE) the Invasive Species Act by the Ministry in humans and dogs in Canada, which is

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 151 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

a serious and potentially fatal “parasitic Veterinary C, Bangalore, India, 3Department tumour” originating in the liver. Echinococcus of Veterinary Microbiology, Karnataka multilocularis, the tapeworm responsible for this Veterinary, Animal And Fisheries Sciences infection, normally occurs in wild canids (foxes, University (b), Veterinary C , Bangalore, India, coyotes and wolves) as its definitive hosts and 4Department of Molecular Virology Lab, IVRI, rodents as the natural intermediate hosts. Wild Bangalore, India canids are sources of infection for humans and dogs serving as aberrant intermediate hosts. Echinococcosis, a cyclozoonotic helminthosis Since humans have close interaction with dogs caused by the dwarf dog tapeworm and with the increasing density of wild canids Echinococcus granulosus. It is distributed in urban areas, it is important to adequately worldwide and common in areas where diagnose the infection in the canid definitive hygienic conditions are poor and literacy hosts so as to prevent or control spread of is low. In India, high prevalence of cystic infection to humans. The gold standard test echinococcosis has been reported but even (adult cestode recovery) for the detection of though dogs are disseminators of infection E. multilocularis is laborious and fatal for the as definitive hosts, very few studies have canid; hence, there has been a shift to molecular conducted. Therefore, the present study was methods of diagnosis. Fecal, heart blood, undertaken to detect the copro-DNA of E. chest fluid, and adult taeniid cestodes were granulosus in dogs by PCR in Karnataka state, recovered from two hundred coyotes trapped South India. Totally 300 faecal samples, which in Saskatchewan winter of 2018. We compared includes 100 samples of dogs collected during prevalence based on adult cestode recovery post-mortem examination and 200 from (73%) with taeniid eggs on fecal flotation and stray dogs were screened by microscopy. centrifugation (27%), and two new copro-PCR Out of 100 dogs examined on necropsy, only assay (73%). Our findings suggest that PCR ten were found positive for E. granulosus based methods offer promising results for worms along with nineteen showed mixed detection of intestinal infections with taeniid infection with Ancylostoma caninum, cestodes in both wild and domestic canids. Dipylidium caninum, Toxocara canis and Finally, it is important to detect cases of AE in Taenia hydatigena. Among 200 faeces of stray dogs as rapidly as possible for a more favorable dogs screened, forty-nine showed eggs of clinical outcome. Therefore, we will also test Ancylostoma caninum, Taeniia sp. Dipylidium coyote blood samples with a novel serological caninum, Toxocara canis, Spirocerca lupi, test for canine AE to ensure that the test Trichuris vulpis, Diphyllobothrium latum does not detect adult cestode infection. It is and Isospora canis. Since, the morphology important to be able to distinguish the two of taeniid eggs including echinococcus possible states in the canid hosts, as one has resembles each other and it’s very difficult to public health implications and the other has differentiate by microscopy alone, the positive animal health implications, and both require samples were subjected to Copro-PCR for prompt intervention. species specific identification of E. granulosus eggs from Taeniia species of dogs using two sets of E. granulosus specific primers of OA40.07 Screening of dogs for Echinococcus Eg 1f, Eg 1r and JB 3f, JB 4.5r. The Eg 1f, Eg granulosus infection by Copro-Polymerase 1r primers yielded a single amplicon of 255 Chain Reaction bp whereas, JB 3f, JB 4.5r yielded 440 bp amplicon which were specific to E. granulosus Dr. Ananda Konanur Javaregowda1, Dr. Placid in twenty-one dog samples. The Copro-PCR Eugin D’Souza2, Dr. Shrikrishna Isloor3, Dr. was found more sensitive in detection and Suryanarayana VVS4 differentiation of E. granulosus from Taeniia 1Department Of Veterinary Parasitology, sp. of dogs and can be used effectively as a Karnataka Veterinary, Animal And Fisheries confirmatory diagnostic tool for detection of Sciences University (b), Veterinary C, E. granulosus in dogs at field level. Shimoga, India, 2Department Of Veterinary Parasitology, Karnataka Veterinary, Animal And Fisheries Sciences University (b),

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 152 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA41 Biology and Pathology of GI three Alberta farms. Survival of C. oncophora Nematodes in Ruminants was proportionally greater than O. ostertagi. Although less than 5% of L3 present on July 10, 2019, 16:30 - 18:00 the pasture in the fall were able to survive Plenary Hall, Madison Ballroom (ABCD), Level 4 the subsequent winter, these were able to infect the tracer calves and establish spring infection. OA41.01 Demonstration of Overwintering of Ostealgia Ostertagi and Cooperia Oncophora These results to date suggest that L3 Larvae on Pasture in Western Canada overwintered larvae play an important role in the epidemiology of GIN infection of beef Dr. Tong Wang1, Ms. Dawn Gray2, Dr. Libby cattle in western Canada and needs to be Redman1, Dr. Russell Avramenko1, Dr. Janneke considered when designing strategic parasite Wit1, Ms. Cailey Patullo1, Ms. Bruna Salles1, Dr. control regimes. John Gilleard1, Dr. Doug Colwell2 1University Of Calgary, Canada, 2Agriculture and Agri-Food Canada (AAFC), Canada OA41.02 Outbreak of Calf Sudden Death Associated with Strongyloides Papillosus Gastro-intestinal nematodes (GIN) have Infection in a Western New York Dairy Farm a negative economic impact on beef production in pastured cattle. Current control Manigandan Lejeune1, Toby Pinn-Woodcock1, recommendations in western Canada are Belinda Thompson1, Gerald E. Duhamel1 based on the assumption that the cold 1Department of Population Medicine & winters in this region (lowest temperature Diagnostic Sciences, College of Veterinary -30 ) prevent overwintering of infective Medicine, Animal Health Diagnostic Center, larvae (L3) on pastures. However, there is Cornell University, United States no published℃ information on the ability of cattle GIN species to overwinter on pastures Strongyloides papillosus are slender hair- in western Canada. Consequently, we are like worms that have a predilection for investigating the overwintering capability of the proximal small intestine of ruminants bovine trichostrongylid L3 larvae on western worldwide. These worms have a unique life Canadian pastures. cycle in which the parthenogenetic females are the only adult stage in the host. They shed A study was conducted on three organic larvated eggs in feces and susceptible hosts farms over two consecutive years. Two are infected percutaneously by the filariform experimental approaches were taken: First, L3. Strongyloides papillosus are generally environmental samples (grass, feces and soil) considered nonpathogenic, as apparently were collected before and after each winter healthy animals tolerate heavy infection. followed by L3 enumeration in the laboratory. Although there are sporadic reports of major The species composition of L3 populations outbreaks with sudden death of calves from was quantified using nemabiome sequencing, East Asia, similar outbreaks have not been a novel deep amplicon sequencing approach. reported in North America. In this report we Second, L3 overwintering was further document an outbreak of sudden death of investigated using tracer calves, a more calves on a NY dairy farm attributed to S. sensitive approach. Parasite-free tracer calves papillosus infection. In early September 2018, were placed on pasture for 3-4 weeks in 15 calves died over a period of 14 days. Calves spring to allow ingestion of overwintered L3 were recumbent with signs of respiratory from pasture before adult worm burdens were distress prior to death, or found dead. All determined for each GIN species. Detailed female calves also had precocious udder weather data at farm level was monitored by development. Necropsy and histopathologic solar powered weather stations. We found findings revealed severe mammary congestion that a proportion of both C. oncophora and and ductal hyperplasia without other O. ostertagi, but not H. placei, L3 larvae significant lesions. Testing for various viral survived on pasture over the winter on all and bacterial agents yielded inconclusive

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 153 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

results. An ancillary finding of 18,800 EPG of In CO -depleted rumen fluid exsheathment S. papillosus in a fecal float from a dead calf of H. contortus could not be achieved, raised suspicion of a helminth-associated whereas₂ O. ostertagi, T. circumcincta and outbreak. Subsequent fecal testing of 7 other O. leptospicularis all showed some degree calves yielded EPG ranging between 1,370 of exsheathment (respectively 46 %, 22 % and 8,300. Shortly after deworming with and 15 %). These findings were confirmed in Dectomax and extensive cleaning of the barn, a CO -free artificial buffer. Interestingly, the death loss stopped abruptly and the udder ability to exsheath in the absence of CO was enlargement resolved. A similar mortality dependent₂ on the composition of the buffer, event on this dairy had occurred the previous indicating the involvement of co-factors.₂ year (July- August, 2017) resulting in the sudden death of 30 calves. Ionophore toxicity Overall, even though these species all exsheath was suspected but feed ionophore levels in the rumen and reside in the abomasum, were within safe limits. Unfortunately, parasite there appear to be significant differences in testing was not performed in 2017. This report their response to exsheathment triggers. The highlights the importance of including S. data suggest an important role for both heat papillosus in the list of differentials for sudden shock and CO in vivo, but in vitro their effects death in young calves. are dependent on the nematode species as well as the surrounding₂ medium.

OA41.03 Nematode Exsheathment 2: Abomasal Species Differ in Their Response OA41.04 Studies on Gastrointestinal to Exsheathment Triggers Nematodes of Australian Alpacas

Dr. Kiliana Bekelaar1, Dr. Tania Waghorn1, Dr. Mr. Harun Rashid1, Dr. Abdul Jabbar1 Michael Tavendale1, Catherine McKenzie1, Dr. 1Melbourne Veterinary School, The University Dave Leathwick1 of Melbourne, Australia 1Agresearch Ltd, Palmerston North, New Zealand Gastrointestinal nematodes (GINs) can cause significant economic losses in alpacas and Infection of ruminants by gastrointestinal llamas. Very little is known about prevalence nematodes is initiated by the exsheathment of GINs, control practices and the use of of the infective third stage larvae. Both anthelmintics. This study aimed to (i) assess carbon dioxide (CO ) and heat shock have the worm control practices used by Australian been shown to play crucial roles in triggering alpaca farmers (ii) assess the prevalence of Haemonchus contortus₂ exsheathment in vitro. GINs and (iii) assess the efficacy of commonly This study set out to investigate how relevant used anthelmintics against GINs of alpacas. these triggers are in rumen fluid and whether An online questionnaire survey was conducted this data can be extrapolated to other species. to assess current worm control practices. A longitudinal and cross-sectional study was In CO -saturated rumen fluid, all species conducted on 13 and 91 selected alpaca farms, showed an efficient exsheathment response respectively, located in different climatic (> 80 ₂% in under 4 hours) following exposure zones of Australia. For the longitudinal study, to heat shock, which was significantly higher a total of 1,692 faecal samples were collected compared to slow temperature changes. In and analysed using faecal egg counts (FEC) the artificial buffer, the effect of heat shock and faecal cultures. The mean faecal egg was species-dependent. For H. contortus count was 167 eggs per gram (epg), with a and Ostertagia leptospicularis the response highest burden of 15,540 epg for strongyle was similar to that in rumen fluid, but type nematodes. In the cross-sectional study, exsheathment of Ostertagia ostertagi and the mean egg count was 291 epg of 1545 Teladorsagia circumcincta was significantly samples analysed. Over one hundred of lower and/or slower, and with no benefit of gastrointestinal tracts were also examined heat shock. to determine the worm burdens and their spectrum. Total worm counts revealed the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 154 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

mean worm burden as 1,291, with a maximum (P=0.1). At the necropsy, abomasum tissue count of 29,000 worms in one alpaca. The samples were collected for histological and main five genera/species identified were immunohistochemistry analysis. A positive Camelostrongylus mentulatus, Haemonchus correlation was found between CD4+ and + contortus, Trichostrongylus spp., Cooperia T cells (r= 0.714; P= 0.04), suggesting that spp. and Nematodirus spp. Finally, the both cell populations could participate in theγδ efficacy of commonly used anthelmintics was early immune response. Moreover, we found assessed by faecal egg count reduction test. an association between the number of + In addition, above three studies, a molecular T cells and eosinophils (r= 0.600; P=0.05); diagnostic kit has been developed which can this association was even stronger in theγδ identify seven different nematodes of alpacas resistant group (r= 0.900; P= 0.037), but from faecal DNA. Results of this study showed absent in the susceptible animals, suggesting that shared-GINs are prevalent in Australian that the activation of this mechanism could alpacas and they are resistant to most widely play an important role in the resistance to the used anthelmintics. This study provides infection. On the other hand, the susceptible invaluable information on the prevalence of group showed a negative correlation between GINs of Australian alpacas, control practices globule leukocytes and + T cells (r= -0.812; and efficacy of commonly used anthelmintics P=0.05) but also slight with CD4+ T cells which could be used to develop control (r= -0.800; P=0.1). Therefore,γδ it is possible strategies against GINs in alpacas. that in the susceptible group the immune response was not strong enough to activate the recruitment of a higher number of globule OA41.05 Differences Within Churra Breed leukocytes to protect against the infection. Sheep in the Early Immune Response to the Infection by Teladorsagia Circumcincta OA42 Diagnostic Techniques Verónica Castilla Gómez De Agüero1, Maria Martinez Valladares1, Elora Valderas García1, July 10, 2019, 16:30 - 18:00 Beatriz Gutierrez Gil1, Juan José Arranz1, Jorge Breakout Room 2, Hall of Ideas E&H, Level 4 F. González2, Julia N Hernandez2 1Universidad de Leon, Leon, Spain, 2Universidad de Las Palmas de Gran Canaria, OA42.01 The Babesia Caballi Spherical Spain Body Protein 4 (SBP-4) Is Recognized by Antibodies in Sera from B. Caballi Infected In the present study, the mechanisms by Equids in Egypt and Has Potential for which Churra breed sheep can present Developing Novel Serological Diagnostic different resistance phenotypes were studied Method in adults during the early stage of the infection by Teladorsagia circumcincta. For Dr. Mona Mahmoud1, Dr. Omnia Kandil1, Dr. that, 12 animals were classified as resistant Nadia Abu El Ezz1, Dr. Seham Hendawy1, (6) or susceptible (6) to the infection by Bassma Mahmoud1 T. circumcincta based on their cumulative 1National Research Centre, Veterinary faecal egg count after an initial experimental Research Division, Parasitology and Animal infection. A negative correlation was found Diseases Department, Dokki, Cairo, Egypt between the cumulative FEC at the end of the infection and the level of IgA in serum Equine piroplasmosis is caused by two intra- at day 3 pi (r=-0.764; P= 0.004), when IgAs erythrocytic haemoprotozoan, Babesia caballi reached the highest values in the resistant (B. caballi) and Theileria equi (T.equi), in most group. Sheep were dewormed and were tropical and subtropical areas of the world infected again to be slaughtered at day 7 where competent tick vectors are present. post infection. At slaughter, level of IgA in A competitive inhibition ELISA diagnostic serum and gastric mucus was higher in the kit based on a monoclonal antibody to resistant group although only showing slight Bc48 (79/17.18.5), reactive with a member of significant differences in serum samples the Babesia sp. rhoptry-associated protein

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 155 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

(RAP)-1 family lacked the ability to detect against Sarcoptes scabiei mites. Motile stages specific antibodies of B. caballi strains in (larvae, nymphs and adults) of S. scabiei mites Egypt, Israel and South Africa. The Spherical were collected from experimentally-infected Body Protein 4 (SBP-4) was had shown an pigs maintained at the veterinary College of excellent performance when used as a new Alfort, France. Petri dishes filled with Columbia serological antigen in several distinct ELISA agar supplemented with pig serum were used diagnostic assay for B. bovis. This study for the bioassays. Each molecule to be tested describes investigations aimed to defining the (beauvericin versus dimpylate and ivermectin SBP4 of B. caballi as an alternative specific as positive controls) was incorporated into antigen that could be used for the detection the medium following the method described of B. caballi strains circulating in Egypt. by Brimer et al. (1993, 1995) with slight The B. caballi recombinant SBP-4 (r-SBP4) modifications. For each molecule, the efficacy protein derived from an Egyptian isolate of of 3 different concentrations (10, 100, and 1000 B. caballi, was purified by immunoaffinity and g/g) was evaluated. All tests were carried immunologically tested. The purified r-SBP4 in quintuplicate with 10 mites/plate at room shows a single 37 kDa band upon coomasie temperature.µ Plates were examined after 1, 2, 3, blue staining of SDS-PAGE gels, which 4, 5, 6, and 24h for survival assessment. Mites reacts with anti-HIS antibodies in Western were considered dead when no movement blot analysis. The pattern of reactivity of the occurred under the microscope. Final data of purified r-SBP4 against antibodies in the sera all treatments were analyzed by Kaplan Meier of equids infected with T. equi and B. caballi survival curves. The present study clearly from an equine population was studied. No demonstrated the activity of beauvericin cross-reactivity was detected when using against motile stages of S. scabiei. After 1h of T. equi positive serum, suggesting that the exposure, the mortality rate was 8%, 20% and r-SBP4 reacts specifically with B. caballi 56% at 10, 100, and 1000 g/g, respectively. infected sera. A total of 191 equids sera were After 6h of exposure, the mortality rate was tested by B. caballi- IFA slides of VMRD Inc. 42%, 86% and 94% at 10, µ100, and 1000 g/g, and a novel SBP-4 I-ELISA. IFAT slides detect respectively. The concentration effect was B. caballi specific antibodies in Egyptian notable at 10 g/g and the mortality recordedµ equine’s samples with low incidence (38.21 %) with ivermectin was significantly higher then in contrast to SBP-4 I-ELISA (61.78%). Hence, that recordedµ with dimpylate and beauvericin. r-SBP4 could be considered as a candidate The survival capacity seems to be different for developing a reliable serological test that according to the evolutive stage of the mites. can be used to detect B. caballi infection in Larvae and nymphs exposed to beauvericin Middle East and Africa. were killed more rapidly than adults.

OA42.02 In Vitro Evaluation of the OA42.03 Improved Patient Care by Activity of a Mycotoxin Produced by the Combining Diagnostic PCR Panels With Fecal Entomopathogenic Fungus Beauveria Antigen and Fecal Flotation Tests in Dogs Bassiana Against Sarcoptes Scabiei With Diarrhea

Charbel Al Khoury2, Dr. Nabil Nemer2, Prof. Christian M. Leutenegger1, Marko M. Estrada1, Jacques Guillot1 M. Alexis Seguin 1Veterinary College of Alfort, France, 2Holy 1Idexx Laboratories, Inc, United States Spirit University of Kaslik, Lebanon Diarrhea is a very common problem in dogs The entomopathogenic fungus Beauveria presenting to veterinary practices with GI bassiana is known to produce beauvericin, a clinical signs. Diagnosing the etiologic cause secondary metabolite with many biological of diarrhea in dogs and selecting the proper effects including insecticidal/acaricidal, treatment at presentation is challenging due antitumor, antibacterial, and antifungal to the many potential causes for an infectious activity. The objective of this study was to GI syndrome. evaluate the in vitro efficacy of beauvericin

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 156 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Real-time polymerase chain reaction (PCR), is widely prevalent and the larval stages also known as quantitative PCR (qPCR), of Dirofilaria, Dipetalonema and Brugia offers improved sensitivity, specificity, safety, species are identified as causative agents speed and scalability compared to other fecal among canines. Although, canine filariosis tests alone for the detection of pathogens is widespread in the temperate and tropical in fecal samples. Improved, rapid detection countries of the world, only few reports are of the often multifactorial causes of canine available in India. Accurate identification diarrhea may allow the initiation of early of canine filarid nematodes is important, etiologic treatments based on the detected because of zoonotic concerns and therapeutic pathogen(s) that are causing disease. This implications. Presently, there is no conclusive will help prevent unnecessary antibiotic use, data available on confirmatory diagnosis facilitate improved management practices, of D. repens in canines and its zoonotic minimize the severity of outbreaks, and implication in Karnataka state. In the present improves pet owner satisfaction. In addition, study, a total of 100 canine blood samples early etiologic treatment reduces the suspected for microfilariosis were collected potential of zoonotic pathogen transmission from different regions of Karnataka. Thirty- to humans, other pets or food animals. eight samples were found positive for In this study, fecal flotation, fecal antigen microfilaria by modified knott’s method ELISA (Fecal DxTM) for ascarids, hookworms, (MKM) and quantitative buffy coat (QBC). and whipworms were combined with a Morphologically, the microfilariae were comprehensive canine diarrhea RealPCRTM unsheathed with blunt head and a tapering panel. Of 157 included samples, 51% tested tail. The nerve ring and excretory cell at the positive for at least one infectious agent by excretory pore region of the microfilaria qPCR. More than 90% of all positive samples could be well appreciated and tail was long were positive for one or more pathogens with hook like posterior end. The micrometry by qPCR, with the majority (82%) positive of microfilaria recovered by the modified by only qPCR. Fecal flotation detected a knott’s method had length of 334.9±4.573 m gastrointestinal pathogen in 15.5% of samples. and width of 6.862±0.133 m. Based on the The Fecal Dx panel was positive for one morphometry, the microfilariae were identifiedµ or more parasites in 4.8% of samples. The as D. repens. To confirm µthe species, the addition of qPCR allowed the detection of 8 samples were subjected to polymerase chain times more infectious agents. reaction (PCR) targeting the cytochrome oxidase subunit I (COI) genes of Dirofilaria This study highlights the benefits of a repens. The amplicons were sequenced and syndromic qPCR panel directed at common analysed. The microfilariae recovered from gastrointestinal pathogens in combination with canines of different regions of Karnataka fecal antigen testing and the fecal flotation were confirmed as D. repens based on PCR tests for the evaluation of canine diarrhea. targeting COI genes. The phylogenetic analysis of the COI gene nucleotide sequence obtained in the present study showed no OA42.04 Phenotypic and Genotypic more than 95% homology with D. repens Characterisation of Canine Filarid sequences existing for comparison. Nematodes of Karnataka, India

Ansar Kamran C.3, Sumana K.2, Malatesh D S1, Ananda K Javaregowda1, Manjunath Reddy G. B.2, Lavanya K. V.3 1Veterinary College, Shimoga, Kvafsu, India, 2ICAR-NIVEDI, India, 3Veterinary College, India

Filariosis is one of the major parasitic diseases of animals and human beings caused by a small group of filarid nematodes throughout the world. Canine filariosis

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 157 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA42.05 Contemporary Status of Equine OA42.06 Detection of Blastocystis Mixed Trypanosomosis in Punjab, India Subtype Infections in Dairy Calves Using Next Generation Amplicon Sequencing Lachhman Singla1 1Department Of Veterinary Parasitology, Guru Dr. Jenny Maloney1, Dr. Monica Santin1 Angad Dev Veterinary and Animal Sciences 1USDA, ARS, EMFSL, United States University, India Blastocystis is a ubiquitous intestinal parasitic Equine trypanosomosis or Surra elicited by protist found in humans and many animals the first pathogenic obligate, eukaryotic, worldwide and is transmitted through fecal intercellular haemo-flagellate protozoan contamination of food or water. Nine of parasite Trypanosoma evansi, is an 22 Blastocystis subtypes have been found unsystematically reported disease of equids in both humans and animals, suggesting in India. The dearth of information regarding zoonotic transmission. Epidemiological data the epidemiological status, role of hemato- on Blastocystis in food animals is limited, and biochemical indices including oxidative stress within-host Blastocystis diversity remains indicators on pathogenesis and the need of largely unexplored. Therefore, a method to field oriented diagnostic test prompted us to study intra-host Blastocystis communities undertake this comprehensive cross-sectional using next generation amplicon sequencing survey, using a combination of parasitological (NGS) was developed and compared to (classical) and sero-molecular diagnostic Sanger sequencing. Fecal samples from cattle techniques together with analysis of risk factor were screened with a PCR that amplifies a associated with them, to unravel the exact fragment of the SSU rDNA gene followed position of epidemiological status of equine by direct Sanger sequencing. If a mixed trypanosomosis covering all equid species infection was suspected, amplicons were from different agro-climatic zones of Punjab. cloned. Seventy-five samples were analyzed The investigation established the endemic by NGS using the same SSU rDNA gene stability of Surra in all agro-climatic zones of region. Sanger sequencing indicated mixed Punjab. The risk factors evaluated indicated infections in 18 of the 75 samples tested, that the donkey/mules, unorganized farms, but only three were successfully confirmed commercial purpose equids and farm without through cloning. NGS identified 49 mixed application of any fly repellent/insecticides infections and revealed 14 subtypes, ten were more vulnerable to get infected from T. previously reported (ST-1 toST-5, ST-10, ST- evansi. Oxidative stress parameters indicated 11, ST-14, ST-17, and ST-21) and four novel that though microscopically positive animal (named ST-23 to ST-26); ST-1, ST-2, and ST-11, (patent infection) were more vulnerable, were not observed by Sanger sequencing. however, latent infection (PCR positive) was Subtypes 1 through 5 are potentially zoonotic, also producing pathogenicity by induced and one or more were found in 79% (59) of anemia and stress. Thus, diagnosis of latent the specimens using NGS. Subtype 3, the infection is important to save the animal from most common subtype found in humans, was induced stress and spread of infection to other found in 37% (28) of specimens tested by susceptible animals. As all the samples positive NGS and in only four specimens using Sanger. by microscopy and PCR were also positive by Blastocystis mixed subtype infections may LAMP assay, indicated LAMP assay to be a be far more common than previously thought promisingly sensitive and specific technique due to the limitations of current detection for the diagnosis of T. evansi under isothermal methods. Better characterization of within- conditions in field situations. Mapping of host Blastocystis subtype diversity in infected endemic area based on GIS predictive humans and animals is needed to improve our prevalence can act as alarming forecast model understanding of Blastocystis epidemiology. for establishment of control programs. The role of cattle as reservoirs of infection for humans and other animals through either direct contact or contamination of food and water should be explored.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 158 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA43 Equine Parasites testing for parasite stages, and were further analyzed for bacteria, bacterial toxins and July 10, 2019, 16:30 - 18:00 viruses by PCR. All animals were categorized Breakout Room 3, Hall of Ideas F&I, Level 4 for their helminth egg shedding potential as high, moderate or low according to AAEP Guidelines. Each individual infectious agent OA43.01 Cross-Sectional Analysis of was then compared to the helminth burden Infectious Disease Agents in Relation to present in the sample to test for an association, Helminth Parasites of Healthy Equidae in an using Chi-squared or Fischer’s Exact Test. This Exhibition Setting study provides initial baseline data on the role that helminth burden may play in modulating Ms. Hayley G. Hofmar-Glennon1,4, Ms. Nicole the occurrence of infectious agents in healthy M. André1, Dr. Alison Stout2,5, Dr. Renata equid populations. Ivanek2, Dr. Gary R. Whittaker1,4, Dr. Laura B. Goodman2,3, Dr. Belinda S. Thompson2,3, Dr. Manigandan Lejeuné2,3, Dr. Erin L. Goodrich2,3 OA43.02 Expansive Chemosensory, Vision, 1Department of Microbiology & Immunology, Immunity, and Insecticide Resistance Gene Cornell University, United States, 2Department Families in the Stable Fly: A Functional of Population Medicine & Diagnostic Sciences, Genomics Tale Cornell University, United States, 3Animal Health Diagnostic Center, Cornell University, Pia Olafson1, Joshua Benoit2, Stomoxys United States, 4Master of Public Health Genome Consortium Program, Cornell University, United States, 1U.S. Department of Agriculture - Agricultural 5Graduate Program in Biomedical & Biological Research Service, Livestock Arthropod Pests Sciences, Cornell University, United States Research Unit, United States, 2Department of Biological Sciences - McMicken College of Helminth infections are known to modulate Arts and Sciences, University of Cincinnati, the host immune response, which is United States thought to either protect or injure the host against other infectious agents. In equines, Stable flies are a global pest of livestock cyathostomins (small strongyles) are production, having economic and social considered the most common helminths, impacts on affected communities. Integrated however, their role in modulating host immune pest management tools are currently response is not clearly known. Horses are employed for population suppression, but predisposed to being infected with strongyles increasing worldwide outbreaks require at either high, moderate or low levels. The development of supplementary and/or spectrum of strongyle burden may influence improved control approaches. Understanding the prevalence of infectious pathogens those biological pathways regulating critical (bacterial or viral) in a healthy population. stable fly behaviors would inform such Moreover, testing for infectious disease development. Here, we report the high agents in feces from healthy equids is rarely quality sequencing and community curation performed and it is unclear which pathogens of the stable fly genome. Coupled with may be present in the general population. transcriptome and bacterial community Understanding the prevalence of infectious analysis, this effort revealed expansions in agents in healthy populations would help gene families that may enable the stable fly to determine the relationship between helminth persist in microbe-rich environments, localize burden and the occurrence of infectious hosts, and detoxify xenobiotic compounds. agents. This will also aid in the interpretation These insights enhance our understanding of of diagnostic test results in animals with this important pest and provide a framework confirmed clinical disease. In this study, 100 for identifying novel control targets. fecal samples were collected from apparently healthy equids at an exhibition in NY State. The feces were examined for helminths and other parasites through morphological

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 159 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA43.03 Surveillance of Equine Strongyle OA43.04 Etiology and Treatment of Buffalo Infections and the Efficacy of a Larvicidal Fly-Associated Lesions Dose of Fenbendazole in Horses Across the United States Dr. Peter James1, Ms. Hayley Kidd2, Dr. Con Constantinoiu3, Prof. Michael McGowan2 Dr. Brian Herrin1, Dr. Duane Chappell2, Dr. 1University Of Queensland, Australia, Bryant Craig2, Dr. Craig Barnett2, Dr. Earl 2University of Queensland, Australia, 3James Gaughan2, Dr. Wendy Vaala2 Cook University, Australia 1Kansas State University, United States, 2Merck Animal Health, United States Buffalo flies (Haematobia exigua), which are very closely related to horn flies (Haematobia Equine strongyles are a common parasite irritans) and are sometimes considered a of grazing equids, and the small strongyles sub species, are a major pest in Australia’s are considered to be the parasite around northern cattle industries. Buffalo flies (BF) which all adult horse deworming protocols are consistently ranked by northern cattle are built. The current study enrolled 410 producers as amongst their main cattle horses on 27 farms (197 weanlings; 213 health issues with the welfare aspects of BF- horses >1 year old) from across the United associated lesions often key amongst their States, to determine the strongyle burdens concerns. These lesions can vary from dry by fecal egg count (FEC) using the Modified and alopecic to open suppurating wounds, Wisconsin technique, as well as the efficacy found most frequently on the medial canthus of fenbendazole as judged by fecal egg count of the eyes, the neck, dewlap, abdomen and reduction test (FECRT) from horses with flanks of cattle. An unnamed species of filarial >150 strongyle eggs on the first FEC. Horse nematode, Stephanofilaria sp., vectored by owners were also asked to fill out a survey on BF, is frequently found associated with these their normal deworming and management lesions in northern herds. However, in the strategies. Horses under 1 year of age received more southern parts of the Australian BF one dose of fenbendazole (10 mg/kg), and range, where lesions are prevalent and can the older horses received the 5 consecutive be severe, detection of Stephanofilaria has days of treatment at 10 mg/kg. The average not been reported from either lesions or BF FEC for all animals with a strongyle-positive dissections. A similar species, Stephanofilaria fecal was 185 eggs per gram (Range: 1 – 1,286 stilesi, is transmitted by H. irritans, but EPG), with horses older than 1 year of age lesions are usually much less severe, ventral having a significantly higher FEC (244.6 in distribution and most commonly found on EPG) than weanlings (95.28 EPG, p-value < the abdomen or near the udder of dairy cows. 0.0001). Deworming reduced overall strongyle Determining the etiology and relative role egg counts by 71.8%, but the efficacy of of BF feeding and Stephanofilaria sp. in the fenbendazole was highly variable between development and persistence of BF lesions farms (Range: 41.6 – 100%). There was no is critical to determining optimal control and significant difference in reduction of strongyle treatment regimes. A study towards this end egg counts between the drug regimen used will be described. for the older horses versus the young horses (p-value = 0.1345). There were no significant management strategies that effected the OA43.05 Species Composition, Seasonal pre-treatment FEC or percent reduction. Incidence and Relative Abundance of Biting The data highlight the fact that most horses Midges at Horse Boarding Facility Located (398/410; 97%) fall into the low or moderate Near Cedar Key, Florida With a Case of egg shedding category for strongyles. While Sweet Itch fenbendazole treatment was >90% effective in reducing strongyle egg counts at some farms, Daniel Kline1 the variability highlights the importance of 1USDA-ARS, CMAVE, United States FEC and FECRT in the routine care for horses to determine the efficacy of the deworming For the past decade an ongoing research program at each barn. project has been conducted at a horse

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 160 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

boarding facility located near Cedar Key, Dogs were experimentally infested with 50 Florida, on the species composition, seasonal ticks and housed individually for 3 to 6h to incidence, and relative abundance of biting allow tick-attachment. Then dogs were co- arthropod populations. This study site has housed in pairs in biosafety rooms throughout gradually been transformed from a scrub the duration of the experiments. Color coded oak forest into a full fletched horse boarding ticks were counted and recorded for each dog facility with several extensive pastures. at 48h and 96h after each infestation. Changes in fauna and flora HAVE BEEN DOCUMENTED. Various trap types and Our results show that each tick species and sampling techniques have been utilized to gender had different migration behavior. monitor the changes in populations of biting R. sanguineus females migrate in higher midges, blackflies, mosquitoes, deerflies, proportion than males when their gender is the horseflies and stable flies. Since a case only one present in the room. However, when of Sweet Itch has been attributed to the both genders are present in the same room, R. biting of the horse by biting midges, this sanguineus males migrate more than females. presentation will focus on the biting midges. D. variabilis females migrate more than males Over 13 species of Culicoides were collected, whether only one or both genders are present but only six consistently from year to year. in the room. Interestingly, only I. ricinus males Three species seemed to bother the horses migrate. Individually marked ticks also showed the most. Culicoides furens was the main that migration within the same host (on-host nuisance species from late spring through migration) occurs frequently. October. Culicoides missisippiensis was the main pest species from mid-October through In conclusion, our data show species-dependent May. Culicoides insignis was abundant from and high mobility of ticks after experimental Novemeber through December and seemed infestation of dogs. This migration behavior to bother the horses a lot. The details of the may influence the transmission of tick borne Sweet Itch case will be presented. diseases, as pre-feeding of ticks on one dog prior to attachment to a second dog would be expected to accelerate transmission of OA44 Canine Arthropods pathogens to the other dog. This underlines the importance of tick prevention in dogs, to reduce July 10, 2019, 16:30 - 18:00 the risk of exposure for pets and humans. Breakout Room 4, Hall of Ideas G&J, Level 4

OA44.02 Assessment of an Activity Monitor OA44.01 Shall We Go for a Walk? – Tick (Vetrax®) and Effect of Pulicidal (Trifexis®) Migration Behavior on Dogs Treatment on Pruritic Behaviors in Flea-Bite Sensitive Dogs Heinz Sager1, Denis Mosimann2, Regina 1 3 Lizundia , Christian Epe Anthony Rumschlag1, Marcel Sarzen2, William 1Elanco Animal Health, Switzerland, 2Idorsia Ryan3, David Young4 Pharmaceuticals, Switzerland, 3Boehringer 1Elanco Animal Health, United States, 2AGL Ingelheim Animal Health, United States Technology, United States, 3Ryan Mitchell Associates LLC, United States, 4Young The objective of this study was to investigate Veterinary Research Services, United States the migration behavior of different tick species between hosts and within the same Activity monitors have potential for guiding host on experimentally infested dogs. veterinarians and owners to treat and to Different sets of experimental infections track canine responses to treatments. A were conducted using three tick species: study investigated the potential for the Dermacentor variabilis, Rhipicephalus Vetrax monitor to track canine pruritic sanguineus and Ixodes ricinus. The ticks were behaviors in flea infested, flea-bite sensitive color coded to allow identification of the host dogs, including following treatment with origin of the respective males and females. a combination product of spinosad and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 161 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

milbemycin oxime (SMO) (Trifexis). Vetrax felis populations are morphologically and units were attached to the collars of 10 genetically conserved, or whether human- mixed breed dogs on Day -14. On each dog, mediated migration of domestic cats and flea infestations of 100 fleas were placed dogs has resulted in homogenous global on Day -13, 30 fleas on Days -10, -7 and -4, populations. To determine the origin of the and 15 fleas were placed daily on Days 7 to species and to investigated the potential 12 and 21 to 27. Video recordings of dogs drivers behind the establishment of regional were scored for pruritic behavior by blinded cat flea populations, we characterised a global observers over a 4-hour period on Days -14, collection of fleas from cats and dogs across -3 (for randomization), 3, 13, 20 and 27. Dogs six continents. Using a multigene approach were randomized to be either untreated combining two mitochondrial (cox1 and cox2) controls or to receive SMO on Day 0. Fleas and two nuclear (Histone H3 and EF-1a) gene were combed from all dogs and counted on markers, as well as a cox1 survey of 516 fleas Days 5, 14 and 28. Comparisons of video- across 56 countries, we demonstrate out-of- recorded with monitor-recorded pruritic Africa origins for the genus Ctenocephalides behaviors were completed for each dog, and high levels of genetic diversity within C. combining data from each four-hour period felis. We define four bioclimatically limited C. over which observations were made. For felis clusters (Temperate, Tropical I, Tropical individual dogs, correlations between video- II and African) using maximum entropy and monitor-recorded behaviors ranged from modelling. This study defines the global 0.48 (an outlier dog that was compulsively distribution, African origin and phylogenetic paw-licking) to 0.91. Overall mean correlation relationships of global Ctenocephalides was 0.70 (P≤0.0001), or 0.75 adjusting data fleas, whilst resolving the taxonomy of from the outlier dog. No fleas were detected the C. felis subspecies and related taxa. in any SMO-treated dog, while control This study reveals the drivers behind the dogs remained infested (efficacy 100%). establishment and success of the cat flea Relative to controls, based on analysis of as a global parasite. Through synanthropic video observations, in the SMO group there host hitchhiking, the cat flea is successfully were significant reductions in mean pruritic achieving global dominance. behavior duration, overall (P=0.0242) (54.4%) and on Day 27 (P=0.0031) (66.2%). SMO reduces or eliminates the reaction of sensitive OA44.04 Ectoparasite Control on Pets: Who dogs to flea bites arising from challenges Did It, With What and Where? A Comparison through the month following treatment. Between Pet Owners from Two High-Risk Vetrax appears to be suitable for assessing Lyme Regions in the United States pruritic responses to pulicidal treatment of flea-bite sensitive dogs. Dr. Gebbiena Bron1, Dr. Maria del Pilar Fernandez2, Dr. Maria Diuk-Wasser2, Dr. Susan Paskewitz1, Dr. Lyric Bartholomay1 OA44.03 Out-of-Africa, Human-Mediated 1University Of Wisconsin - Madison, United Dispersal of the Common Cat Flea, States, 2Columbia University, United States Ctenocephalides Felis: The Hitchhiker’s Guide to World Domination Many pet owners know that outdoor pets are the proverbial canary in a coal mine for tick Prof. Jan Slapeta1 activity. Dogs and cats readily pick up ticks, 1Sydney School of Veterinary Science, which inspires owners to seek ectoparasite University of Sydney, Australia control measures for their pet. However, the initiation and adherence to recommended The cat flea (Ctenocephalides felis) is ectoparasitic treatment frequencies can be the most common parasite of domestic dependent on the owner’s perceived risk of cats and dogs worldwide. Due to the disease. Lyme disease (LD) and black-legged morphological ambiguity of C. felis and a lack ticks (Ixodes scapularis, also known as deer of — particularly largescale — phylogenetic ticks) are highly abundant in the United data, we do not know whether global C. States Northeast (NE) and Midwest (MW).

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 162 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

As part of a broader human behavioral study participated and 812 cats and 662 dogs were on tick encounters, we asked participants examined. Overall, 28.1% of cats and 14.4% about pet-tick encounters and details on pet of dogs were flea infested. More than 90% tick control, using a smartphone application of the fleas on both cats and dogs were as a survey tool (The Tick App). We aimed Ctenocephalides felis felis. Fleas of the same to understand if there are differences in species from each infested host were pooled. the implementation of tick control on pets DNA was amplified from 470 of the pooled between the MW and NE, and if controls flea samples using conventional PCR, 66 of increase with the owner’s personal protective which (14% ± 95% CI 3.14%) were positive for at measures? The majority of dog owners least one pathogen. Fifty-three (11.3% ± 95% CI treated their dog at least once during the 2.85%) of the pooled flea DNA samples were summer (91.4% of 574 respondents). There positive for Bartonella spp., 35 were from cats was no significant difference in treatment and 4 from dogs. Seventeen of the Bartonella frequencies (never, once, every 2-3 months, spp. samples were found to be Bartonella every 4-6 weeks, more frequent than every 4 henselae, 27 were Bartonella clarridgeiae, 4 weeks) between the NE and MW; about half samples were Bartonella alsatica and one was of the dog owners treated their dog every Bartonella grahamii; 4 samples could not be 4-6 weeks (51.7%). Interestingly, this included identified. Fourteen (3% ± 95% CI 1.53%) of the 17 dog owners who used a product with a flea DNA samples were found to be positive for recommended use frequency of every 8-12 Dipylidium caninum, 10 of which were collected weeks, implying overuse of the product. Dog from cats and one from a dog, the other 3 owners who reported more tick prevention positive flea samples had no host species measures for themselves were also treating record. Only 3 flea samples were positive their dogs more frequently, and dog owners for Mycoplasma haemofelis or Mycoplasma were also more likely to report daily tick haemocanis; 2 were collected from cats and checks of their pet if they had found a tick on one had no host species record. Three flea their pet(s) the previous summer. In summary, samples had were positive for more than one the implementation of tick checks and pathogen. This study highlights the need for ectoparasite control on pets was correlated ongoing flea control, particularly given the to the owner’s experience with prevention relatively high prevalence of Bartonella spp., strategies and finding ticks on pets. which is of concern for both animal welfare and human health.

OA44.05 Pathogens in Fleas Collected from Cats and Dogs: Distribution and Prevalence OA44.06 Tick Biting Trends on Pets in the UK Revealed by Crowdsourced Data: Some Surprises and Implications for Prevention Dr. Swaid Abdullah1, Dr. Chris Helps2, Prof. Séverine Tasker2, Ms. Hannah Newbury3, Prof. Heather Kopsco1, Steven Engborg1, Roland Richard Wall2 Duhaime1, Dr. Thomas Mather1 1The University of Queensland, Australia, 1University Of Rhode Island Center for Vector- 2University of Bristol, United Kingdom, 3MSD borne Disease & Tick Encounter Resource Animal Health, United Kindgom Center, United States

Fleas (Siphonaptera) are the most clinically Citizen science is increasingly utilized to track important ectoparasites of dogs and cats. important vectors of human and companion Rising levels of pet ownership, climate animal disease, providing a scalable, cost- change and globalisation are increasing the effective, and highly-sensitive strategy for importance of understanding the endemicity identifying new foci, changing phenology, and prevalence of flea-borne pathogens. The and associated disease impacts across study recruited veterinary practices around wide geographies. We describe a digital the UK, asked to follow a standardised flea tick surveillance program that provides inspection protocol on a randomised selection photograph-based tick identification and of cats and dogs. A total of 326 practices public health messaging services free to

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 163 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

the public and share important insights into OA45 Ruminant Trematodes II pet tick encounters and pressing tick bite prevention needs. July 10, 2019, 16:30 – 18:00 Breakout Room 5, Meeting Rooms KLOP, Level 4 From 2014-2016, the University of Rhode Island’s TickSpotters program received 17,906 tick photograph submissions from all 50 U.S. OA45.01 Is There Are an Interplay Between states and Canadian Provinces. More than Fasciola Hepatica and MAP Infection in half (55.5%) were from Lyme endemic states, Cattle? but submissions where pet was identified as host were more evenly distributed between Ms. Amalia Naranjo Lucena1, Laura Garza- endemic (49.2%) and non-endemic (50.1%) Cuartero1, Dr. Conor McAloon2, Dr. Alan Wolfe1, states. Pets were identified as hosts in 23% Dr. Guy McGrath3, Dr. David Graham4, José of all submissions while 10% were ticks found Pérez5, Prof. Grace Mulcahy1, Dr. Annetta Zintl1 loose and wandering on humans, pets, or in 1Veterinary Pathobiology Section, UCD School the home. Ixodes scapularis (43.7%) were of Veterinary Medicine, Ireland, 2Herd Health the most common species found attached and Animal Husbandry Section, UCD School to pets, followed by Dermacentor variabilis of Veterinary Medicine, Ireland, 3UCD Centre (26.7%), Amblyomma americanum (6.6%), and for Veterinary Epidemiology and Risk Analysis, Rhipicephalus sanguineus (5.9%). These ticks Ireland, 4Animal Health Ireland, Ireland, were mostly adults (88%), and ticks found on 5Department of Anatomy and Comparative pets had a longer estimated engorgement Pathology, Faculty of Veterinary Medicine, time (median three days) compared to University of Córdoba, Spain humans (median one day). Ticks were spotted year-round; during spring and summer, ticks Co-infection with Fasciola hepatica interacts from pet hosts made up 20.1% and 16.8% of with Mycobacterium bovis infection in cattle, all submissions, respectively, while pet host altering the response to tests for the diagnosis submissions increased to 34.8 and 39.1% of bovine tuberculosis and reducing bacterial during autumn and winter, respectively. load. The importance at an epidemiological scale is still unclear. We hypothesized that F. Crowdsourced data reveal that mostly adult hepatica might also interact with the clinical ticks are spotted on pets, and that they are course and/or epidemiology of infection detected later in the blood-feeding process with Mycobacterium avium paratuberculosis than humans, putting pets at heightened (MAP), the cause of Johne’s disease, and risk for tick-borne disease transmission. The used epidemiological and histopathological doubling of reports of ticks found on pets methods to investigate. during autumn and winter may reveal a critical knowledge gap regarding seasonal activity A database of MAP ELISA test results and of Ixodes scapularis, vector of Lyme disease, F. hepatica liver scores reported in abattoirs providing an opportunity for prevention- from 62,447 cattle in Ireland was analyzed. education to pet owners. A model for Johne’s disease infection was developed using both herd and environmental data. We found that Johne’s positive herds were less likely to be positive for F. hepatica infection ( 2 =13.895, df=1, P<0.002, OR=0.75, 95% CI: 0.62–0.9). However, multivariate analysis showedχ that the main risk factors for Johne’s disease were herd size, soil, and rainfall with F. hepatica co-infection ranking at position 17.

In addition, histological sections were obtained from the ileum of 17 animals with clinical Johne’s disease, with and without

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 164 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

concurrent or recent F. hepatica infection. transmission risk of F. hepatica in this area. Lesions were classified, and numbers of The method was able to detect up to 10-4 immune cells and apoptosis quantified by pg genomic DNA in a 25 L PCR reaction immunohistochemistry. The two animals with system even with high concentrations of snail current liver fluke infection only developed DNA, and no cross reactionμ being observed diffuse intermediate lesions, whereas from the genomic DNA of Paramphistomum animals which were either not infected with cervi, Clonorchis sinensis, Orientobilharzia, F. hepatica or had historic infections also Metorchis orientalis, Dicrocoelium chinensis. developed multifocal, diffuse multibacillary Of 409 snail samples, the overall F. hepatica and diffuse lymphocytic lesions. Moreover, prevalence is 43.76%. However, the F. hepatica there was less evidence of apoptosis in infection of snails was 92.75 % in the Tibetan animals with concurrent fasciolosis than in Autonomous Prefecture of Gannan, while no animals which had no concurrent liver fluke snail was detected in 216 samples collected in infection (P=0.026). Hui Autonomous Prefecture of Linxia. The nest- PCR was firstly used to detect the infection Our results show that while F. hepatica of F. hepatica in snail. It is a novel, useful and infection appears not to be a major factor in convenient method with high sensitivity and the epidemiology of Johne’s disease, it may specificity. This is the first report about the affect lesion development in the intestine epidemiological surveillance of F. hepatica and hence its clinical course. Further in vitro infection in Radix cucunorica in northwestern studies are warranted to investigate these China, which will help to evaluate the effects in more depth. transmission risk of F. hepatica in this area.

OA45.02 Development of a Nest-PCR for OA45.03 Molecular and Morphological the Detection of Fasciola Hepatica in the Characterization of Liver Fluke Intermediate Intermediate Snail Host, Radix Cucunorica, Host Lymnaeids (Gastropoda: Lymnaeidae) and the Evaluation of Transmission Risk in Snails From Selected Regions of Okavango Northwestern China Delta of Botswana, Kwazulu-Natal and Mpumalanga Provinces of South Africa Si-yang Huang1 1College of Veterinary Medicine, Yangzhou Prof. Samson Mukaratirwa1, Ms. Pulane University, China Malatji1, Prof. Jennifer Lamb1 1University of KwaZulu-Natal, School of Life Fasciolosis, a foodborne zoonotic disease, Sciences, South Africa caused by Fasciola hepatica, which is considered an important problem for human The study aimed at identifying populations health and livestock husbandry development. of Lymnaeidae snails from selected sites of Snails are intermediate hosts of F. hepatica, the Okavango Delta (OKD) in Botswana, and the epidemiological surveillance of snails sites located in the KwaZulu-Natal (KZN) can evaluate the transmission risk of this and Mpumalanga provinces of South Africa disease in human and livestock. The aim of using a combination of shell morphology this study was developing a nest-polymerase and molecular approaches. Lymnaeidae chain reaction (nest-PCR) to detect the F. snails were collected from 8 locations from hepatica infection in Radix cucunorica, a the Okavango delta in Botswana, 9 from prevalent intermediate host of this parasite KZN and one from Mpumalanga provinces in northwestern China. The nest-PCR was and were identified based on phylogenetic used to amplify a 208 bp fragment of the analysis of the internal transcribed spacer second internal transcribed spacer (ITS- (ITS-2). Principal component analysis (PCA) 2) of F. hepatica with two pairs of primers, was performed to assess if Lymnaeidae respectively. To evaluate the transmission risk species determined by analysis of the ITS-2 of this disease, 409 snail samples collected marker can be distinguished through shell from different areas of Gansu province morphometry measurements. Analyses based China, were used to detect and analyze the on the ITS-2 marker identified the presence

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 165 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

of a well-supported Radix clade containing related to the genera Metorchis, Clonorchis Radix auricularia, R. (Lymnaea) natalensis and and Opisthorchis. Recently, a high prevalence R. rubiginosa, which were not well resolved. of Amphimerus infection in humans, cats, Experimental samples from the OKD and KZN and dogs had been demonstrated in a present in this clade were referable to these tropical Pacific region of Ecuador. Hence, species. An unidentified experimental taxon we determined and characterized the entire from the OKD formed a well-supported sister mt genome sequences of adult liver flukes, clade to the Radix clade, although it was morphologically identified as Amphimerus, not possible to identify it. Galba truncatula collected in the endemic region of Ecuador, was well supported in a sister relationship and examined its phylogenetic relationships to a well-supported Pseudosuccinea with flukes in the Opisthorchiidae family columella clade which included samples from using Bayesian inference (BI) based on the Mpumalanga and KZN provinces of South concatenated amino acid sequences and Africa. We observed that P. columella shared partial cox1 sequences. The complete mt the same habitats with R. natalensis and genome sequence (15, 151 bp in length) of R. auricularia in KZN. Shell morphometric the Amphimerus sp. contains 35 genes, studies were not able to distinguish among including 12 protein-coding genes (PCGs, the above-mentioned genera and species. without atp8), two rRNAs (rrnL and rrnS) The study further identifies the species which and 21 tRNAs, lacking trnG. The gene content are likely to co-exist in the same environment and arrangement of the Amphimerus sp. and this information will be of use to those mt genome was similar to those of other designing control programs for fasciolosis. trematodes in the Opisthorchiidae family. This is the first study reporting the presence Genetic distances between Amphimerus sp. of R. auricularia in the OKD of Botswana and and other genera in Opisthorchiidae were KZN province of South Africa. rather high, ranging 26.86% to 28.75% at nucleotide level and 29.37% to 31.12% at amino acid level. Phylogenetic analysis placed the OA45.04 Characterization of the Ecuadorian Amphimerus within the branch of Mitochondrial Genome Sequences of the Opisthorchiidae, but it was not gathered with Liver Fluke Amphimerus (Trematoda: flukes from Opisthorchis. Our results indicate Opisthorchiidae) From Ecuador and that the disputable liver fluke Amphimerus Phylogenetic Implications from Ecuador does not belong to the genus Opisthorchis, and that it should be assigned Dr. Jun Ma1, Dr. Jun-Jun He1, Miss Cheng- under the valid genus Amphimerus. This Yan Zhou1, Dr. Miao-Miao Sun1, Dr. William study provided a new genetic marker for Cevallos2, Prof. Hiromu Sugiyama3, Prof. Xing- future studies on taxonomy and molecular Quan Zhu1, Prof. Manuel Calvopiña4 epidemiology of Opisthorchiidae trematodes. 1State Key Laboratory of Veterinary Etiological Also, the determination of mt genome Biology, Key Laboratory of Veterinary sequences of the Ecuadorian Amphimerus Parasitology of Gansu Province, Lanzhou has implications for diagnosis, drug design, Veterinary Research Institute, Chinese control and prevention of amphimeriasis. Academy of Agricultural Sciences, China, 2Carrera de Medicina, Facultad de Ciencias Médicas, Universidad Central del Ecuador, OA45.05 Global Serum Proteomic Changes Ecuador, 3Department of Parasitology, in Water Buffaloes Infected with Fasciola National Institute of Infectious Diseases, Gigantica Japan, 4OneHealth Group, Carrera de Medicina, Facultad de Ciencias de la Salud, Dr. Jun-Jun He1, Dr. Fu-Kai Zhang1, Dr. Rui-Si Universidad de las Américas (UDLA), Ecuador Hu1, Prof. Hany Elsheikha2, Dr. Zhao-An Sheng3, Prof. Wei-Yu Zhang3, Dr. Wen-Bin Zheng1, Prof. Amphimerus Barker, 1911 is a liver fluke Xing-Quan Zhu1 infecting several animal species and 1Lanzhou Veterinary Research Institute, humans. Being a digenetic trematode of the Chinese Academy of Agricultural Sciences, Opisthorchiidae family, Amphimerus is closely China, 2School of Veterinary Medicine and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 166 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Science, University of Nottingham, Sutton ruminants worldwide. With the emergence of Bonington Campus, United Kingdom, 3College resistance to anthelmintic treatments, efforts of Animal Science and Technology, Guangxi have focused on the development of a safe University, China and effective vaccine against this pathogen. Various studies have explored the use of many The liver fluke Fasciola gigantica modulates different antigens from this parasite within several signaling pathways in infected buffaloes a range of hosts. Although promising results to facilitate its survival and establishment of have been obtained, further efforts to increase persistent infection, while the buffalo host the level and consistency of protection would activates immune responses to counter be useful. Furthermore, it is proposed that the infection. Isobaric Tags for Relative the parasite may use decoy epitopes in an and Absolute Quantitation (iTRAQ)-based effort to misdirect the host immune response. quantitative proteomic analysis of the serum of We hypothesised that lymph node samples F. gigantica infected buffaloes can reveal serum from vaccinated and infected animals could protein changes that are linked with F. gigantica be used to identify protective and non- infection process. In present study, a total of protective T cell epitopes within antigens of 313, 459 and 399 proteins were identified; interest, with the ultimate aim of developing of these 92, 93 and 138 were differentially smart designer vaccines to protect ruminants abundant proteins at 3, 42 and 70 days post against fasciolosis. infection (dpi), respectively. Differentially abundant proteins were involved in key Lymph node cells were isolated from biological processes, such as the complement non-vaccinated infected cattle and from system, coagulation and platelet activation, vaccinated infected cattle. These cells were lymphocyte binding on liver epithelium and stimulated with overlapping peptides from lysozyme hydrolysis. Other important markers two antigens, FhCL1 and FhCL3 that have of infections were six proteins, which were been shown to be useful in vaccinates. Cell significantly upregulated in infected serum at all proliferation was measured using the BrdU three time points after infection. These findings assay and IFN production quantified by provide novel insight into the serum proteomic ELISA. Cattle vaccinated with FhCL1 and signature of buffaloes during the course of F. FhCL3 demonstratedγ higher levels of cell gigantica infection. proliferation than cattle vaccinated with FhCL1 alone while, in general, vaccinated cattle demonstrated higher cell proliferation than OA45.06 Identification of T Cell Epitopes control non-vaccinated infected cattle. IFN Within Cathepsins L1 and L3 From Fasciola production was found to be stimulated by a Hepatica Associated with Cell Proliferation region within the signal peptide sequence ofγ and Production CL1 (amino acids 3-15) along with the region within the active enzyme (amino acids 200- Dr. LaurenIFNγ M. O’Neill1, Dr. Laura Garza- 280) which has also been shown to be a B cell Cuartero1, Dr. Thomas Geurden2, Dr. Suman M. epitope. This work demonstrates the potential Mahan3, Dr. John M. Hardham3, Professor John of sub-unit vaccines to target T-cell responses P. Dalton4, Prof. Grace Mulcahy1 towards specific T-cell epitopes and this, 1School of Veterinary Medicine, University together with identification of protective College Dublin, Ireland, 2Veterinary Medicine B-cell epitopes, could aid in the development Research and Development, Zoetis Inc, of protective effective anti-fluke vaccines. Belgium, 3Veterinary Medicine Research and Development, Zoetis Inc., United States, 4School of Biological Sciences, Medical Biology Centre, Queen’s University Belfast, United Kingdom

Fasciola hepatica, the common liver fluke, has a geographically-widespread distribution, resulting in significant economic losses in

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 167 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA46 Gastrointestinal Protozoa in strengths and weaknesses between the Ruminants different loci and between short- and long- read metabarcoding approaches is discussed. July 11, 2019, 8:30 - 10:30 Plenary Hall, Madison Ballroom (ABCD), Level 1 OA46.02 A New Real-Time PCR Method for Identification of Two Pathogenic Eimeria OA46.01 Comparing Different Metabarcoding Species in Sheep, E. Ovinoidalis and E. Approaches to Investigate Eimeria Species Crandallis Diversity and Community Structure in Cattle Lea Bordes1, Dr. Christelle Grisez2, Francoise Dr. Libby Redman1, Dr. Berit Bangoura2, Dr. Prevot2, Didier Marcon3, Philippe Jacquiet2 Bruna Palmeira1, Ms. Nicollette Shaw3, Ms. 1UMR INRA/ENVT 1225 IHAP, France, 2National Jill De Rijke3, Ms. Rebecca Chen1, Dr. Matt Veterinary School of Toulouse, ENVT, France, Workentine1, Prof. John Gilleard1 3Domaine expérimental Bourges-La Sapinière 1Faculty of Veterinary Medicine University of INRA, France Calgary, Canada, 2Department of Veterinary Sciences, University of Wyoming, United Coccidiosis in lambs, caused by intestinal States, 3University of Waterloo, Canada parasites of the Eimeria genus, has high medical and economic impacts throughout Bovine coccidiosis, caused by protozoal the world. Diagnosis of lamb coccidiosis parasites of the genus Eimeria, is an is currently performed in laboratory by important cause of clinical disease and sub- microscopic examinations of fecal material clinical production loss in cattle with control but this method is time-consuming. being largely dependent on the routine Moreover, the species identification, based on administration of anticoccidial drugs. Whilst microscopy, requires experienced personnel E. zurneii and E. bovis are considered to be for accurate identification as oocysts of the most important, over 20 different species many different species are difficult to of Eimeria have been described in cattle distinguish morphologically. As an alternative and relatively little is known regarding their to morphological approach, a real-time pathogenesis, prevalence, drug sensitivity or Polymerase Chain Reaction using Taqman interactions with other pathogens. We have probes has been developed to quantify all undertaken a metabarcoding approach to Eimeria oocysts in the sample and to identify explore bovine Eimeria species diversity and and quantify more specifically oocysts of community structure. Paired-end short read Eimeria ovinoidalis and Eimeria crandallis, sequencing (Illumina Miseq) was used to the species associated with clinical ovine target three variable regions of the SSU 18S coccidiosis. As available genomic data rRNA coding sequence as well as the ITS-1 are scarce, a strategy of sequencing was rDNA region. The full length SSU 18S rRNA performed on different Eimeria spp oocysts, coding sequence was also targeted with isolated by micromanipulation. Sequences long-read sequencing (PacBio Sequel SMRT). of 18S were used to designing primers and The different approaches were compared probes for three PCR test: a Pan-Eimeria test to evaluate their ability to successfully for estimate the total number of oocysts, estimate the species proportions on a variety a Eimeria crandallis test and a Eimeria of different samples. These included mock ovinoidalis test for the amount of oocysts of communities of known Eimeria species each pathogenic species present in a fecal proportions and field samples from beef sample. The Locked Nucleic Acid technology and dairy cattle across Western Canada in was used to increase the specificity of which ~100 sporulated oocysts had been the PCR reactions. Thanks to a modified morphologically identified per sample. pUC57 plasmid from Escherichia coli and The species proportions determined by an estimation of the average of gene’s copy metabarcoding were broadly similar to the number within and between species, the morphological data but differences between standard curve accurately quantifies all the various methodologies occurred. The oocysts in the sample and the proportion of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 168 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

each pathogenic species. This new diagnostic pups is analysed by ELISA. The presence tool could be used in sheep farms for of anti-gp45 antibodies in the mouse monitoring ovine coccidiosis and for an early colostrum would support the possibility to detection of pathogenic species. passively immunize newborn calves against cryptosporidiosis. Preliminary data suggest that C. parvum gp45 is a promising vaccine OA46.03 Cryptosporidium Parvum gp45 candidate and support further investigation in Protein Is a Promising Vaccine Candidate to a cow/calf model of infection. Prevent Bovine Cryptosporidiosis

Dr. Karine Sonzogni-Desautels1,2,3, Dr. Timothy OA46.04 New Leads and Drug Targets in Geary3, Dr. Momar Ndao1,2,4 the Zoonotic Cryptosporidium Parvum by 1Research Institute of the Mcgill University Phenotypic Drug Screening Health Centre, Canada, 2Infectious Diseases and Immunity in Global Health Program, Haili Zhang1, Zi Jin1, Fengguang Guo1, Guan Canada, 3Institute of Parasitology of McGill Zhu1,2 University, , Canada, 4National Reference 1Department of Veterinary Pathobiology, Centre for Parasitology, Canada College of Veterinary Medicine & Biomedical Sciences, United States, 2Institute of Zoonosis, Bovine cryptosporidiosis is a scourge in Jilin University, China the dairy industry because newborn dairy calves can get infected as soon as they are Among the 26 or more Cryptosporidium born and can die during the first two weeks species, C. parvum is the major zoonotic of age due to Cryptosporidium parvum- agent infecting humans and other related severe diarrhea and dehydration. mammals. Currently, there is a lack of fully Our goal is to immunize pregnant cows to effective drugs to treat cryptosporidiosis transmit neutralizing antibodies through in both humans and animals. In the anti- colostrum to the newborn calf. We examined Cryptosporidium drug discovery campaign, several candidates for immunization and we developed an in vitro assay suitable selected C. parvum gp45 as the most for high-throughput screening of anti- promising antigen. We established by liquid cryptosporidial activity. Using this assay, we chromatography/tandem mass spectrometry have screened 1200 existing drugs and 800 that C. parvum gp45 is secreted during natural products with defined molecular excystation of sporozoites from oocysts. structures, and discovered a number of hits Confocal microscopy confirmed that gp45 is or leads with excellent efficacy against the expressed by C. parvum during excystation, growth of C. parvum in vitro (i.e., low nM most likely to promote parasite adhesion to sub-μM EC50 values) and large safety to host intestinal cells. We are currently intervals (i.e., much higher cytotoxicity TC50 investigating the ability of recombinant values than EC50 values). These hits and C. parvum gp45 to bind intestinal cells in leads are serving as templates for developing vitro. C. parvum-infected interferon gamma more efficacious and safer compounds. receptor knock-out (IFNgammaR-KO) mice Additionally, the phenotypic screening data present clinical signs of cryptosporidiosis also revealed and confirmed several drug similar to those of Cryptosporidium spp.- targets in Cryptosporidium. Some of the infected newborn calves. Adult IFNgammaR- targets are being characterized and explored KO mice were immunized with recombinant for target-based drug discovery against gp45 and challenged with C. parvum and cryptosporidiosis. These findings provide vaccination reduced intestinal parasite burden basis for developing more selective and by up to 80%. To determine if neutralizing potent inhibitors, and for studying the mode/ antibodies against C. parvum gp45 can be mechanism of action of leads on the parasite. transmitted by colostrum, we immunized female IFNgammaR-KO mice twice and bred them after the last immunization. Colostrum extracted from the stomach of the newborn

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 169 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA46.05 Cryptosporidium Parvum Animal OA46.06 Evaluating the “Protobiome”: A Model in Neonatal Calves Novel Diagnostic Tool for the Sustainable Control of Protozoan Parasite Communities Dr. Debora Smits1, Dr. Erik van Engelen1, Dr. in Ruminants Deterink-Damhuis1, Dr. W.A.J.M. Swart1, Dr. Lisette Kastelein1, Prof. A.G.J. Velthuis2 Umer Chaudhry1, Qasim Ali2, Imran Rashid2, 1GD Animal Health, The Netherlands, 2Cow Muhammad Zubair Shabbir2, Mike Evans1, Manager, The Netherlands Kamran Ashraf2, Ivan Morrison1, Liam Morrison1, Neil D. Sargison1 A Cryptosporidium parvum model was set 1University Of Edinburgh, United Kingdom, up for in future efficacy studies of veterinary 2University of Veterinary and Animal sciences products. Nine calves were included from Lahore, Pakistan which six were inoculated and three were negative controls. The calves were orally Piroplasmosis is caused by tick-borne inoculated with 10 (one animal 10 ) haemoprotozoa of the genera Theileria and commercially available viable oocysts at Babesia, that impose a serious impact on day of enrollment. ⁶Health parameters⁷ were animal production and human health. While scored twice a day and feces was sampled multiple haemoprotozoan species can infect a once a day (mixed). Calves were weighted single host, there is a lack of reliable molecular once a week. Health parameters included: diagnostic tools with which to understand overall clinical appearance, body temperature, the composition of these complex parasite feed (milk) intake and signs of dehydration. communities. Haemoprotozoa vary in their In daily feces, consistency was scored on epidemiology, drug sensitivity, pathogenicity a 3 point scale, and fecal volume, fecal dry and interaction of co-infected species in the matter content and fecal oocyst count were host, while common features include that determined. animals become the persistent carriers of these pathogens after primary recovery from All calves from the inoculated group shed infection and can play a significant roles as C. parvum oocysts and no control calves a reservoir hosts. In the present study, we shed C. parvum oocysts. When estimated developed for the first time, the use of deep with PCR, shedding started 3 to 5 days after amplicon sequencing using an Illumina Mi- inoculation and stopped between 14 and 17 seq platform to identify haemoprotozoan days after inoculation. All calves showed signs communities and to establish the concept of of diarrhea, both clinically and estimated a “protobiome”. Haemoprotozoa of ruminants by dry matter content. Based on dry matter were used to develop the concept, because content, the duration of diarrhea differed mixed species infections are common. between the calves from 1 to 10 days. When a First, four phenotypically verified species score of 2.5 per day was regarded as diarrhea, of Theileria and Babesia were used to all inoculated calves but no control calves prepare mock pools with random number of experienced diarrhea for more than three parasites, and amplified with four different days. All inoculated calves showed some signs numbers of PCR cycles to assess the species of general clinical illness, ranging from mild in representation bias. Second, we evaluated the some calves to more severe in others. threshold of the deep amplicon sequencing With this model, generating shedding of assay for each of the four species present at C. parvum and diarrhea in all calves while different levels of parasitemia and to confirm evoking only moderate symptoms of general the accurate relative quantification of all illness, we have a model that is suitable for four species. Finally, we applied the assay testing Investigational Veterinary Products. to the field samples to afford insight of the species composition of haemoprotozoan communities in small and large ruminants in the Punjab province of Pakistan. The “protobiome” concept has a wide range of potential applications in veterinary and human research including responses to drug

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 170 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

treatment, parasite epidemiology/ecology, OA47 Wildlife Helminths parasite interactions during mixed infections and parasite control strategies. July 11, 2019, 8:30 - 10:30 Breakout Room 2, Hall of Ideas E&H, Level 4

OA46.07 Cryptosporidiosis in Pre-Weaned Graded Murrah Buffalo Calves of Coastal OA47.01 Occurrence, Prevalence and Districts of Andhra Pradesh State, India Intensity of Internal Parasite Infections of African Lions (Panthera Leo) in Enclosures at Prof. Ravipati Venu1, Dr. V.C.S. Naik1 a Recreation Park in Zimbabwe 1Sri Venkateswara Veterinary University, India Dr. Whatmore Tagwireyi1, Dr. Norman Cryptosporidium is an obligate intestinal Mukarati2, Prof. George Vassilev2, Dr. Michael coccidian parasite, which infects a wide range Tavengwa2 of hosts. In the present study, Cryptosporidium 1University Of Pretoria, South Africa, in pre-weaned buffalo calves of coastal 2University of Zimbabwe, Zimbabwe districts (Guntur, Krishna, West Godavari and East Godavari) (Krishna-Godavari river A coprological survey was conducted to basin) of Andhra Pradesh was studied. Dung determine the types, prevalence, and intensity samples (n=505) from pre-weaned buffalo of infection of internal parasites in a population calves under one month old were screened for of captive African lions at a recreational Cryptosporidium infection. In modified cold game park in Zimbabwe. Fecal samples were strong Ziehl-Neelsen (mZN) staining technique collected over a 4-month period from each 54 (10.69 %) dung samples were found positive of 30 lions (55%) out of 55 animals held. The for Cryptosporidium oocysts, whereas in FEA samples were examined using flotation and sedimentation coupled mZN staining method sedimentation techniques as well as larvae detected 51 (12.14 %) positives. Randomly identification. The overall prevalence of selected 267 dung samples were subjected to helminth infections was 100% (30/30), and 80% nested PCR for detection of Cryptosporidium (24/30) of fecal samples also were positive for and 71 (26.59 %) positives were recorded. protozoan parasite forms. Eggs of Ancylostoma Based on the results of mZN staining method, spp. were found in the feces of 23 (76.7%) lions, the prevalence of Cryptosporidium infection Physaloptera sp. in 14 (46.7%) lions, Toxascaris was higher (15.30 %) in the Godavari river basin leonina in 13 (43.3%) lions, Toxocara cati in 12 (West Godavari and East Godavari districts) (40%) lions, and Gnathostoma spinigerum and and lowest in the Krishna river basin (Guntur Toxocara canis in 2 (6.7%) lions. Furthermore, and Krishna districts) (4.91 %). Nested PCR eggs of Cylicospirura subequalis, Gnathostoma detected 14.91 (17/114) and 35.29 percent spp., Lagochilascaris major, Acanthocephalan, (54/153) of Cryptosporidium infection in Linguatula spp. and larvae of Aelurostrongylus Krishna and Godavari river basins, respectively. spp. were identified in the feces of one lion. DNA sequence analysis revealed that, the Oocysts of five apicomplexan parasites and majority of the buffalo calves has C. bovis cysts of one mastigophoran protozoan parasite infection, followed by C. ryanae and C. parvum were recorded, namely, Cystoisospora leonina infection. The zoonotic species, C. parvum was in 11 (36.7%) lions, Cystoisospora spp. in 9 recorded both from the Krishna river basin (30.0%) lions, Cystoisospora felis in 5 (16.7%) and the Godavari basin areas of the Andhra lions; Toxoplasma-like spp. in 5 (16.7 %) lions, Pradesh state. In conclusion, this is the first and Giardia spp. in 8 (26.7%) lions. The majority report on the Cryptosoridium in pre-weaned of lions (28/30) showed mixed infections buffalo calves of the Krishna-Godavari river with different internal parasite. Eimeria spp. basin region of Andhra Pradesh state. oocysts, identified were spurious and probably originated from prey species. Among the parasites identified were some of zoonotic importance that have health implications for at- risk personnel and visitors who get into contact with the animals.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 171 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA47.02 Clinical Crenosomosis Lungworm OA47.03 Fecal Examinations to Inform Infection in a Black Bear (Ursus Americanus) Deworming Protocols of Raccoons (Procyon lotor) Entering a Rehabilitation Center in Ms. Haifaa Mahjoub1, Nicole Murphy1, Dr. Upstate New York Paula-Marie Mather2, Dr. Spencer Greenwood1, Dr. Gary Conboy1 Dr. Araceli Lucio-Forster1, Dr. Bridget Barry2, 1University of Prince Edward Island, Canada, Dr. Dwight Bowman1 2Maritime Animal Hospital, Canada 1Cornell University College of Veterinary Medicine, United States, 2Cornerstone An orphaned 9-month-old underweight black Veterinary Hospital, United States bear was evaluated at a wildlife rehabilitation facility for chronic cough and wheezing. Fecal A local veterinarian working in collaboration samples were submitted to the AVC Diagnostic with a local wildlife rehabber in Upstate New Services for parasitological examination York state, was interested in developing a for detection of GI helminths. Baermann tailored deworming protocol for raccoons examination was performed due to the detection (Procyon lotor) entering the facility. To of first-stage nematode larvae (L1) on centrifugal determine the presence, identity and flotation; large numbers (>8000 L1/gm) of L1 prevalence of parasitic infections, fecal samples were recovered and identified as Crenosoma from raccoons under his care were examined spp. based on morphology. Three species have over a 1-year period (September 2017 through been reported from black bears (Crenosoma September 2018). A total of 37 samples were petrowi, Crenosoma potos, Crenosoma vulpis). collected from raccoons of various ages, Based on larval length measurements, the sometimes housed as litters. Fecal samples L1 were tentatively identified as C. petrowi. were stored under refrigeration, and processed The length of L1 ranged from 253.4 to 276.7 within 7 days of collection. Each sample µm. Further molecular characterization using was examined by 1.18 spg zinc sulfate and Polymerase Chain Reaction (PCR) and DNA 1.3 spg sugar centrifugal flotation. Parasites sequencing of the small subunit (SSU) RNA were detected in 30 of the 37 fecal samples gene and two regions of the large subunit (LSU) processed. These infections consisted of three rRNA gene did not match any submissions in genera of protists and at least four genera of GenBank, but were most similar to Crenosoma nematodes and one trematode; no cestode mephiditis. The only three sequences available infections were diagnosed. Single parasite from the GenBank database are Crenosoma infections were diagnosed in 18 samples, while vulpis, Crenosoma mephitidis, and Crenosoma 10 had 2 parasites present, and 2 samples striatum. There is a paucity of molecular had more than 2 parasites detected (3 and data for members of the genus Crenosoma. 5, respectively). Protistan infections were Unfortunately, no sequence data on Crenosoma the most common, followed by nematode, petrowi or Crenosoma potos are currently and trematode infections. The most common available in GenBank. Therefore, lungs were parasite detected was Cryptosporidium, and obtained from black bears during the following the most common co-infection was with spring hunting season. Analysis of partial SSU lungworms. Details of the findings of the and LSU rRNA gene sequences from adult-male- investigation and potential contributing factors C. petrowi recovered from the black bear was will be presented and discussed. identical to that obtained from the bear cub L1 confirming the morphological identification as C. petrowi. Following treatment with Panacur® OA47.04 Occurrence of a Zoonotic (fenbendazole), clinical signs resolved. This is the Tapeworm, Taenia Crassiceps (Zeder, first report of clinical chronic respiratory disease 1800) Rudolphi, 1810 in a Muskrat (Ondatra due to Crenosoma infection in a black bear. Zibethicus) in New York State, USA

Amira Abdu1, María J Forzán1, Manigandan Lejeune1 1Department of Population Medicine & Diagnostic Sciences, College of Veterinary

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 172 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

Medicine, Animal Health Diagnostic Center, Heterobilharzia americana is a trematode Cornell University, United States parasite of mammalian wildlife in the southern United States that is related to the human Taenia crassiceps is a widely distributed schistosome, Schistosoma mansoni. The parasite across the Holarctic, cycling most common definitive host is the raccoon between canine and rodents as definitive (Procyon lotor), but it is found in many other and intermediate hosts, respectively. In hosts such as coyote (Canis latrans), opossum contrast to other Taenia species, T. crassiceps (Didelphis virginiana), nutria (Myocastor has proliferative cysticercus that develops coypus) and less frequently in dogs or other asexually by budding, thus ingestion of domestic animals. one or a few eggs can result in massive infection. In this report, we describe a case In the mammalian host, adult worms live of cysticercosis attributable to the taeniid in the mesenteric veins, and eggs migrate cestode T. crassiceps in the carcass of an through the intestinal wall to be excreted adult male muskrat from New York State. in feces. Eggs can enter the circulation Gross inspection revealed numerous round to and become trapped in other organs often square pearly white soft structures (1-2 mm in causing granulomatous inflammation. The diameter) present on the serosal surface of all impact of this infection in dogs can range abdominal viscera and the peritoneal surface from asymptomatic to fatal, and clinical of the diaphragm. These structures were findings may include hemorrhagic enteritis, identified as taeniid cysticerci and based on diarrhea, weight loss, hepatic disease, and the characteristic rostellar hook morphology hypercalcemia. were confirmed as that of Taenia crassiceps. Histopathological examination revealed In a historical report, H. americana was found cysticerci with distinct rostellar hooks, lateral in a small number of Florida panthers (Puma suckers, and numerous calcareous corpuscles. concolor coryi), an endangered subspecies of Remnants of hooks associated with small puma restricted to a small geographic area in mineral deposits were occasionally observed Florida, USA. In this study, archived tissues in the adipose tissue of the retroperitoneal collected opportunistically from 18 panthers space. Molecular investigations with PCR (12 male, 6 female, age range 4 months-4 amplification using a universal primer set years, median age 2 years) necropsied targeting a partial sequence of mitochondrial between 2004-2016 were evaluated cytochrome c oxidase subunit 1 (COX1) gene histologically for H. americana infection. of taeniid yielded amplicons which were Sections of small intestine and liver were Sanger sequenced and blasted to confirm examined for all 18 animals, and additional T. crassiceps. This reported case emphasize tissues including pancreas, lung, and kidney the zoonotic threat pose by T. crassiceps in were examined in some cases. Despite poor New York State and the need for continued fixation and advanced autolysis of many surveillance in domestic dogs to reduce the tissues, trematode eggs were observed in risks to humans. the small intestine of 5 animals, and also in the liver of one of those. Eggs were most commonly observed in the intestinal mucosa OA47.05 Investigating the Potential Impact and were not associated with inflammation of Heterobilharzia Americana on Endangered in most cases. Molecular techniques were Florida Panthers (Puma Concolor Coryi) attempted to confirm the identity of parasites in selected tissues from multiple animals. To Dr. Carolyn Hodo1, Dr. Jessica Y. Rodriguez2, preserve this endangered population, it is Dr. Bambi Clemons3, Dr. Mark W. vitally important to determine the impact of Cunningham3, Dr. Karen F. Snowden1 infectious diseases such as H. americana. 1Texas A&M University, College of Veterinary Medicine, United States, 2ZOETIS, United States, 3Florida Fish and Wildlife Conservation Commission, United States

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 173 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA47.06 Mammomonogamus N. Sp. From The introduction of the bank vole (Myodes the Nasal Cavity of River Otter (Lontra glareolus) to Ireland in the 1920’s presents Canadensis) in Canada a unique opportunity to investigate how bio-invasions alter the parasitic dynamics Dr Gary Conboy1, Nicole Murphy1, Dr Spencer of invasive and native species. Such models Greenwood1, Dr Pierre-Yves Daoust1 are required due to the association between 1Atlantic Veterinary College-Upei, Canada translocations of hosts and parasites and emerging infectious diseases. Bank voles and The nasal cavities of 308 river otter (Lutra native woodmice (Apodemus sylvaticus) were canadensis) obtained as a by-product after trapped in Ireland at the point of introduction pelting from trappers in Eastern Canada and of the vole, at the expansion front and in areas the Maritimes were examined for helminth where only woodmice are found. Parasite infection. The trematode, Clinostomum surveys were carried out and 16SrRNA marginatum was found in 4.2% (13/308) sequencing was used to detect major genera of the otter with a mean infection intensity of pathogenic bacteria. Results demonstrate of 2.4 flukes/infected animal (range = 1-16 that the influence of the bio-invasion on flukes). A nematode identified as a species pathogen dynamics is dependent on where of Mammomonogamus (Family Syngamidae; the animals are located across the expansion Subfamily Syngaminae) was recovered from range. Overall voles trapped in Ireland have 8.8% (27/308) of the otter. The mean infection less helminth parasites than those elsewhere intensity was 6.6 worms/infected animal in Europe. Also at the expansion front they with a range of 1-22 worms recovered. In have significantly less parasites than at the most instances adult female worms were core population. Helminth parasites of the recovered with male worms attached (=“Y” native woodmice are also diluted, with a worms). Female worms were 11-22 mm in significant decrease in Brillouin’s Index of length (Mn=15.4 mm) and red in color; male Diversity, where the vole has been established worms were 2-5 mm in length (Mn=3.7 mm). longest. Metagenomic results show voles Adult worms had a large buccal cavity with and mice share pathogenic bacteria in their longitudinal ribs visible on the inner wall. Eggs endemic range (France) and the pathobiome in the uterus of the female lacked opercula of mice from France and Ireland is dominated and the rays in the male genital bursa were by the same pathogenic bacteria. However rounded. All previously described species voles in Ireland have a unique pathobiome of Mammomonogamus have been reported to that of voles in their endemic range. This from tropical or subtropical regions of the unique pathobiome may be related to the world infecting the larynx, nasal cavities and changes in the host parasite community. middle ear in cervids, felids, ruminants and the elephant. Endemic Mammomonogamus infection has not been reported in river otter or OA47.08 The Role of Wildlife in Parasitic in a temperate region previously. Zoonoses: The Case of Philippine Wildlife

Dr. Vachel Gay Noreen Paller1 OA47.07 The Unseen Side of a Bio- 1University of the Philippines, Philippines Invasion: A Unique Spatiotemporal Study of Infection Dynamics in a Natural Large-Scale Wildlife ecosystem has been subjected to Perturbation Experiment environmental and anthropogenic impacts such as deforestation and climate change. Peter Stuart1, Karen Loxton2, Nathalie Many wildlife species are being displaced Charbonnel3, Maxime Galan3, Heikki and unable to adapt to environmental Henttonen4, Joeseph Jackson5, Colin Lawton6, alterations leading to biodiversity loss in areas Celia Holland1 of urban development. However, some wild 1Trinity College Dublin, Ireland, 2University animals are attracted to human habitations College Cork, Ireland, 3CBGP, France, 4LUKE, due to abundant food supply and presence Finland, 5University of Salford, United Kingdom, of structures where animals could shelter. 6National University of Ireland, Ireland Macaques are common sights in some urban,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 174 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

agriculture, and tourist areas in South East metabolism, interactions with host immune Asia, including the Philippines. The same is cells, and xenobiotic defense mechanisms. true for rodents, wild cats, wild pigs, and other Numerous studies report the maintenance wild animals in some residential, agriculture of adult Ascaris suum employing a variety and agroforestry areas in the country. of nutrients, media, and environmental Unfortunately, some of these wildlife species conditions. However, no studies have are also hosts for a number of parasites of determined the requirements of the equine public health and veterinary importance. Due ascarid, Parascaris spp. Helminth longevity to the complexity of many parasitic life cycles is traditionally assessed on an alive/dead involving several host species, the interactions basis, and some reports subjectively assessed between wild animals, domestic animals and viability. Currently, none of the published humans could enhance transmission of these methods allow for a viability assessment of parasites. Our recent studies confirm the adult Parascaris spp. while accommodating presence of five simian Plasmodium spp. in their large size. The aims of this study was macaques, eight parasite species in Palawan to establish the in vitro requirements of native leopard cats, 12 helminth parasite Parascaris spp., and to develop an objective species in wild rats, and 15 parasite species in viability assessment method. In total, 1045 wild native pigs. In addition, hemoparasites worms were maintained over 212 cultures. of birds and reptiles were recorded. Moreover Worm longevity and viability was assessed. risk factors of zoonotic transmission due to Worms obtained from naturally infected animal-human conflicts will be discussed. foals at necropsy were maintained in culture Since more than 75% of human diseases are of flasks containing 200 mL of culture media. zoonotic origin, it is important to understand Various media, nutrients, and environmental the dynamics of wildlife, domestic animal, conditions were examined. A scoring system and human nexus, as well as conduct more (0-7) based on muscle tone and motility was interdisciplinary researches, to include socio- used to assess worm viability. Worms were economic factors, for control strategies of evaluated every 12 hours. The addition of zoonotic transmissions. glucose to ascaris ringers solution resulted in a significantly longer lifespan (84 hours) than any of the other added nutrient supplements, OA48 Equine Ascarids but glucose did not improve viability. Of all the media and nutrient supplements July 11, 2019, 8:30 - 10:30 tested, worms maintained in the Roswell Breakout Room 3, Hall of Ideas F&I, Level 4 Park Memorial Institute-1640 medium had significantly better viability (P<0.0001) and a maximum lifespan of 168 hours. The use of OA48.01 Long Live the Worms: In Vitro a platform rocker also significantly improved Maintenance for Intestinal Stages of viability (P=0.0305). The CO2 incubator did Parascaris Spp. And a Method for Assessing not influence worm longevity or viability. Viability This is the first study to examine the in vitro requirements for maintenance of Parascaris Jessica Scare1, Dr. Ashley Steuer1, Dr. Carrie spp. intestinal stages and to objectively Shaffer1, Dr. Paul Slusarewicz2, Dr. Angela evaluate their viability over time. Mousley3, Dr. Martin Nielsen1 1University of Kentucky, Gluck Equine 2 Research Center, United States, MEP Equine OA48.02 Hatching Protocol and in Vitro 3 Solutions, United States, Queen’s University Exposure to Ivermectin in Parascaris Belfast, School of Biological Sciences, Ireland Univalens

Parascaris spp. is a clinically important Dr. Eva Tydén1, Frida Martin1, Sofia Jonsson1 parasite infecting young horses and has wide- 1Swedish University of Agricultural Sciences, spread anthelmintic resistance. Maintenance Sweden of helminth parasites in vitro facilitates the elucidation of numerous processes, such as The equine roundworm, Parascaris univalens

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 175 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

is a pathogenic parasite of foals and yearlings roundworm Parascaris univalens is an and infection can result in impaired growth increasing threat to equine welfare as and intestinal obstruction. Drug resistance resistance to all available drug classes have to macrocyclic lactones is spread worldwide been found in recent years. Despite the first and in recent years resistance to pyrantel report of treatment failure to ivermectin has been reported in the U.S and Sweden 17 years ago the mechanisms for drug and to benzimidazoles in Australia. Little is metabolism and drug resistance in this known about molecular mechanisms linked species are still largely unknown. The aim of to anthelmintic resistance (AR) and currently this project is to gain knowledge about the no in vitro model is available for P. univalens. molecular mechanisms of drug metabolism The aim of this project is to understand the and resistance in P. univalens. We have molecular mechanisms of AR by developing investigated the expression of a number of an in vitro hatching protocol for P. univalens candidate genes believed to be involved in the and to study gene expression after drug drug metabolism of P. univalens by qPCR. exposure of the larvae. P. univalens eggs were Adult P. univalens were obtained from isolated from foals naturally infected with two 6 month old Icelandic horses that the parasite. The external chitinous layer was had never been treated with anthelmintic removed from unembryonated eggs using drugs. The worms were collected at an 2% sterile sodium hypochlorite solution and abattoir, transported to the laboratory and incubated in 25 ℃ for 10 days for development in vitro exposed to ivermectin, pyrantel of larva. Hatching was induced according or thiabendazole for 24 h. RNA was then to a protocol for Toxocara canis with slight extracted from the anterior end of the worm, modifications of additional three strokes converted to cDNA and used as template for in a glas homogenizer to improve number PCR and quantitative real time PCR. hatched larva. Hatched larva were cultured for 16 days in RPMI (containing 10% FBS 1% Results show that drug metabolizing enzymes Penicillin and Streptomycin, 1% L-glutamine) such as the phase I cytochrome P450 family, in 37 ℃ with 5% CO2. The hatching ratio of phase II glutation S-transferases and UPD- this protocol was approximately 28% and glycosyltransferases and transport proteins the survival rate of hatched larva 85% after such as ABC transporters are expressed. 16 days. In addition, expression of genes Preliminary results also show that phase I and involved in the drug metabolizing pathways phase II enzymes are differentially expressed were studied in hatched P. univalens larva after drug exposure. after 24 h of exposure to ivermectin. Our hope is that this in vitro method will help us gain This is the first time phase I and II drug more knowledge about AR and the genes metabolizing enzymes have been shown to involved in different pathways responding to be expressed in P. univalens. Results from anthelmintic treatment in ascarid worms. The this study will hopefully help us gain more situation with development of resistance to all understanding of the mechanisms of drug avaliable drug classes is an increasing threat metabolism and AR. for the equine industry and animal welfare.

OA48.04 Differential Transcriptome OA48.03 Expression of Drug Metabolizing Analysis of Parascaris Univalens After In Genes in Parascaris Univalens Vitro Exposure to Ivermectin, Pyrantel and Thiabendazole Frida Martin1, Dr. Matthías Eydal2, Prof. Johan Höglund1, Dr. Oskar Karlsson Lindsjö1, Sofia Dr. Frida Martin1, Dr. Oskar Karlsson Lindsjö1, Jonsson1, Dr. Thomas F. Bergström1, Dr. Eva Dr. Matthías Eydal2, Prof. Johan Höglund1, Dr. Tydén1 Tomas F. Bergström1, Dr. Eva Tydén1 1Swedish University of Agricultural Sciences, 1Swedish University of Agricultural Sciences, Sweden, 2University of Iceland, Iceland Sweden, 2University of Iceland, Iceland

Anthelmintic resistance (AR) in the equine Anthelmintic resistance in Parascaris univalens

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 176 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

is an increasing threat to equine welfare as OA48.05 Effect of Macrocyclic Lactones on resistance to all available drug classes have Parascaris Sp. Glutamate-Gated Chloride been found in recent years. Despite this Channels fact, the genetic and molecular background of drug metabolism and resistance in P. Nicolas Lamassiaude1, Dr. Elise Courtot1, Dr. univalens is still largely unknown. The aim Cédric Neveu1, Dr. Claude L. Charvet1 of this study is to gain deeper knowledge 1ISP, INRA, Université Tours, France about drug metabolizing genes in P. univalens. To achieve this a whole genome Parascaris sp. is the largest parasitic approach was applied, which is an unbiased nematode of horse causing digestive and and systematic method to identify genes respiratory disorders to the animal. The involved in different pathways responding to control of equine ascaridiosis relies on anthelmintic treatment. We have used RNA- anthelmintic treatments including macrocyclic Seq to compare the transcriptomes of adult lactones (MLs) as the gold standard. However, P.univalens after in vitro exposure to three control of infestation is increasingly difficult different drug classes. due to the emergence of resistant parasites throughout the world. In the free-living model Adult P.univalens was obtained after slaughter nematode Caenorhabditis elegans, glutamate- from two 6-month old Icelandic horses that dependent chloride channel receptors had never been treated with anthelmintic (GluCls) were identified as the main targets drugs. In the laboratory the worms were in of MLs. However, in Parascaris sp, the mode vitro exposed to ivermectin, thiabendazole of action of MLs remain poorly understood. and pyrantel, each at three different Here we identified the Parascaris sp. GluCls concentrations, for 24 h. RNA was extracted and characterized the effect of a wide range from the anterior end of the parasite and of MLs. three biological replicates per concentration were sequenced using Illumina NovaSeq. Using a candidate gene approach, we identified the orthologs of 6 genes encoding Sequencing of 36 samples (27 exposed and 9 GluCls subunits in Parascaris sp. The controls) generated 20,462,622 to 35,338,763 complete cDNAs encoding these subunits paired-end reads with a length of 100bp per were amplified by PCR and cloned into a sample. A mapping index was created using transcription vector. The corresponding the previously published genome (GenBank: cRNAs were synthesized in vitro and then NINM00000000.1), transcript expression were microinjected into Xenopus laevis oocytes. quantified using Salmon and differential gene Two-electrode voltage-clamp experiments expression analysis was performed using R were performed on recombinant GluCls to package DESeq2. investigate their pharmacological properties. Thus, the expression of a single subunit and Preliminary results show that between 35 combination of different subunits in Xenopus and 474 genes are significantly differentially oocytes allowed us to obtain the functional expressed (p < 0,05) in response to exposure homeric and heteromeric GluCls of Parascaris depending on drug and concentration. Ongoing sp. The receptors were both sensitive to analysis of KEGG pathways will provide insights glutamate and ivermectin but the effect of about the function of these genes and should seven different ML compounds revealed help us gain more understanding of drug striking differences. The physiological function metabolism in P.univalens. and the impact of MLs on these receptors in vivo are in progress.

This study, provides a better understanding of the pharmacology of GluCls as well as the mode of action of MLs in nematode parasites.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 177 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

OA48.06 Ascarids Exposed: In Vitro Drug Gene functionalities are related to muscle Exposure and Gene Expression Analysis of contraction, detoxification and exocytotic Anthelmintic Naïve Parascaris Spp. events, and regulatory processes. qPCR did not identify any significant differences Jessica Scare1, Dr. Pouya Dini1, Jamie Norris1, between groups, but this may be due to the Dr. Jianbin Wang2, Dr. Ashley Steuer1, Dr. lack of method sensitivity. The results of Kirsten Scoggin1, Holli Gravatte1, Dr. Daniel RNA-seq analysis and gene function warrant Howe1, Dr. Paul Slusarewicz3, Dr. Richard further investigation and their role in ascarid Davis2, Dr. Martin Nielsen1 xenobiotic responses. 1University of Kentucky Gluck Equine Research Center, United States, 2University of Colorado, Department of Biochemistry and Molecular OA48.07 Gastrointestinal and Respiratory Genetics, United States, 3MEP Equine Nematodes in Feral Horses of Sable Island Solutions, United States – A Living Experiment in Equine Parasite Epidemiology Ascarid parasites infect a variety of hosts, and control measures rely heavily on anthelmintics Dr Emily Jenkins1, Amber Lynn Backwell1, raising concerns for anthelmintic resistance. Alice Liboiron1, Julie Colpitts1, Christina Parascaris spp., the equine ascarid, is the Tollett2, Sarah Medill3, Jennifer Bellaw4, only ascarid with documented substantial Todd Shury1,3, David McRuer3, John Gilleard5, drug resistance. Elucidating drug responses Jocelyn Poissant5, Philip McLoughlin1 of susceptible parasites may lead to further 1University Of Saskatchewan, Canada, understanding of anthelmintic resistance 2Queen’s University, Canada, 3Parks Canada, mechanisms, identifying alternative drug Canada, 4Gluck Equine Research Center, targets, and parameters for slowing the United States, 5University of Calgary, Canada development of resistance among other ascarid species. The purpose of this study was Sable Island National Park Reserve is a small to examine the response of anthelmintic naïve sandy island located approximately 275 km Parascaris spp. to in vitro drug exposure using east of Halifax, Nova Scotia, Canada. Horses transcriptomic analyses and quantitative PCR. (about 500 head) are the only terrestrial Adult Parascaris spp. worms were obtained mammals on the island, and survive despite at necropsy from foals born into a herd kept high pasture densities and absence of human without anthelmintic treatment. Adult worms interventions such as supplemental feeding were maintained in Roswell-Park Memorial and deworming. Strongyle fecal egg counts, medium 1640. The drugs employed were monitored in all live horses every summer, oxibendazole (OBZ) and ivermectin (IVM), are high (mean 1500 eggs per gram - EPG), and 10-fold serial dilutions were prepared. vastly exceeding the guidelines to deworm at Optimum drug concentration and exposure values greater than 200 EPG in companion length were determined using a worm viability horses. Larval culture has demonstrated that assessment method. Subsequently, RNA-seq small strongyles (cyathostomes) had the analysis identified genes with significantly highest relative abundance amongst Sable different expression between drug treated horses, followed by the large strongyles and control groups. Finally, a selection of the Strongylus equinus, S. edentatus, and S. significant genes was further investigated vulgaris. In April 2017, we examined 30 using qPCR in a repeat drug exposure trial. natural mortalities and found evidence of Following initial viability assessments, OBZ cyathostomiasis, pathology associated with (10 g/mL) for 24 hours and IVM (1 g/ larvae of S. vulgaris in the cranial mesenteric mL) for three hours were the conditions artery of yearlings, and pancreatic damage employed.μ RNA-seq analysis revealedμ a total associated with larvae of S. equinus. We of 88 transcripts with significantly ( =0.01) also found high prevalence of Dictyocaulus different expression between all drug treated arnfeldi in airways, a lungworm not normally and control worms. The top five genesα thought to circulate in horses in the absence selected for further analysis were snap25, of donkeys, the normal maintenance host. cytochrome p450, kelch10, sup9, and ferm. Ten of 12 (83%) yearlings were infected, and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 178 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

lungworms were also found in 8 of 18 (44%) diminished reproductive capacity. Integrated adult horses. Dictyocaulus larvae were tick management (ITM) strategies have been present in 3 of 31 (10%) of fecal samples developed that have defined tactics including collected from live horses in early August. habitat and wildlife management, fencing, Identification as D. arnfeldi was based on DNA grazing rotations and cattle treatments characterization of the ITS 2 region of both with acaricides. Tick management requires adult and larval nematodes. In April 2018, all producers to gather and physically inspect fecal samples examined from live yearlings animals on a regular basis to determine tick (n=8) had larvae of Dictyocaulus (mean of 17 presence and abundance, and make informed larvae per gram -LPG) while 13 of 36 (36%) decisions regarding management tactics. of adult horses were shedding larvae (mean Animal stress, time, labor, facilities wear and 3 LPG). Transmission of this lungworm is expense are disincentives to ITM adoption. enigmatic, and further investigation is needed Near infrared reflectance spectroscopy (NIRS) to determine significance as a respiratory has previously been used to detect tick pathogen, especially in yearlings which had infested cattle and monitor success of ITM the highest intensities of infection. tactics. Ticks modulate the immune system of cattle in order to obtain a blood meal over a period of days to weeks. This modulation OA49 Ticks on Cattle of the immune system results in cascading effects through the endocrine system to the July 11, 2019, 8:30 - 10:30 digestive system producing changes in fecal Breakout Room 4, Hall of Ideas G&J, Level 4 chemistry that are detectable by NIRS. This presentation will evaluate if NIRS analysis of bovine feces (fNIRS) can discern different OA49.01 Detection of Winter Tick Infested levels of winter tick infestations on cattle, Cattle Using Near Infrared Reflectance further testing the sensitivity and feasibility Spectroscopy of Bovine Feces of fNIRS technology as an alternative method expected to improve ITM adoption, decision- Ms. Samantha Hays1, Dr. Pete Teel1, Dr. Sonja making, and efficacy for tick management. Swiger2, Dr. Jeffery Tomberlin1, Dr. Thomas Hairgrove3, Dr. David Anderson3 1 2 Texas A&M University, United States, Texas OA49.02 Modeling Cattle-Nilgai-Deer A&M AgriLife Research and Extension, United Interactions to Assess Impact of Nilgai on 3 States, Texas A&M AgriLife Research and Cattle Fever Tick Eradication Efforts in the Extension, United States U.S.A.

External parasites annually cost the U.S. beef Kimberly Lohmeyer1, Hsiao-Hsuan Wang2, cattle industry $2.4 billion through the direct William Grant2, Denise Bonilla3, Hallie Hasel4, effects of parasitism, and an even greater Andy Schwartz5, Pete Teel2, Adalberto Perez cost when animal handling and treatment de Leon1 expenses are included. Tick parasitism of 1USDA-ARS, United States, 2Texas A&M range cattle can occur year-round. While University, United States, 3USDA-APHIS- most species are active in spring and summer VS, United States, 4USDA-APHIS-VS, United months, certain species such as the winter States, 5Texas Animal Health Commission, tick, Dermacentor albipictus, are active in United States fall and winter when forage quality and quantity are low. Dermacentor albipictus is Abundant nilgai, Boselaphus tragocamelus, recognized as a one-host tick economically in South Texas pose significant challenges to important in large ungulates and is a known cattle fever tick (Rhipicephalus microplus and vector that has previously been shown to R. annulatus) eradication efforts. Modeling transstadially transmit Anaplasma marginale, cattle-nilgai-white-tailed deer interactions causal agent of bovine anaplasmosis. Direct provides a tool to assess the impact of nilgai production losses accrue from tick parasitism on the efficacy of standard cattle fever tick through bloodloss, irritation, weightloss, and eradication protocols. To meet this need, we

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 179 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

revised our spatially-explicit, individual based, +/- 4.7 kg) at the USDA-ARS Cattle Fever tick simulation model that simulates cattle Tick Research Laboratory, in Edinburg, TX, fever tick population dynamics in response to before, during, and after an infestation with tick control measures in the presence of cattle 5000 R. microplus larvae. Cluster analyses and/or white-tailed deer on hypothetical were conducted using GRAMS IQ for NIR heterogeneous landscapes in southern Texas. spectra in the 576-1126 nm range to test The updated model includes nilgai as an for fecal chemistry changes different from alternative cattle fever tick host species and pre-infestation condition, and coincident refines information on rangeland landscapes with the biological phases of R. microplus in and around the permanent quarantine zone infestation. The first three factors of spectral in south Texas along the U.S.-Mexico border. variation accounted for 87.87% of spectral Information on nilgai life history and ecology, variation among all samples. Factors 1, 2 and including patterns of habitat use and efficacy 3, had F-Ratios for the Reduced Eigenvalues as a tick host were analyzed and used to of 941.59, 387.44 and 221.79, respectively. describe movement rules quantitatively, and Three-dimensional analysis for these 3 to estimate the spatial and temporal dynamics factors shows shifts in sample clusters away of host seeking larvae in the system. Updates from pre-infestation and coincident with on the development of the cattle-nilgai-white- progressive R. microplus blood-feeding and tailed deer model and how it can be utilized post-infestation recovery. We conclude that to adapt cattle fever tick suppression tactics fecal NIRS may provide a tool for detection of utilized by the Cattle Fever Tick Eradication R. microplus-infested cattle. However, further program will be presented. testing is needed to determine the sensitivity of detection on cattle with varying levels of R. microplus burden, and a protocol developed OA49.03 Detection of Fecal Chemistry and evaluated for fecal sampling under field Changes in Cattle Infested With the Southern conditions. Cattle Tick, Rhipicephalus (Boophilus) Microplus OA49.04 Interaction Between a Systemic Mr. Brian Rich1, Dr. Pete Teel1, Dr. Donald Acaricide and Immunological Control of Thomas2, Dr. Jay Angerer1, Dr. Douglas Rhipicephalus (Boophilus) Microplus Tolleson1, Dr. Adalberto Perez-DeLeon3 1Texas A&M University, United States, 2USDA Charluz Arocho Rosario1, Robert Miller2, Pete ARS, United States, 3USDA ARS, United States Teel1, Felix Guerrero3, Adalberto Perez de Leon3 Bovine babesiosis, a highly fatal tick-borne 1Texas A&M, United States, 2USDA-ARS, disease of cattle, was eliminated from the United States, 3USDA-ARS, United States US in the last century through the federal and state Cattle Fever Tick Eradication The southern cattle fever tick Rhipicephalus Program against the cattle fever ticks (CFT) (Boophilus) microplus, causes large economic Rhipicephalus microplus and R. annulatus. losses in cattle production, particularly in The threat to the US cattle industry continues tropical and subtropical parts of the world. through CFT introductions from Mexico. In the United States losses were estimated The standard method to detect infestation to be ~$130.5 million in the late 1800’s before by the Program is physical examination of the eradication program began. Ectoparasites restrained cattle to find CFT. New methods of develop easily in tropics and sub-tropics and detecting CFT-infested cattle could improve are responsible for large economic losses in reliability and reduce animal stress. This the dairy and meat industry. study determined whether changes in fecal chemistry induced by R. microplus infestation The southern cattle tick R. microplus is a was detectable using near-infrared reflectance one-host tick species considered the most spectroscopy (NIRS). Fecal samples were important ectoparasite of livestock in the collected daily from 6 stanchioned Bos taurus world because of its association with high yearling heifers (initial mean weight 163.3 kg financial loss due to direct feeding and in the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 180 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

transmission of the hemoparasites Babesia acaricides are the main strategy of tick bovis, B. bigemina, and Anaplasma marginale, control, however, there are reports of the causative agents of babesiosis and development of resistance from various anaplasmosis, respectively. parts of the world. Thus present study was designed to check the cypermethrin Rhipicephalus microplus has a high potential resistance against cattle tick at three different for population growth due to its relatively Govt. livestock farms and their surrounding short life cycle and preference for cattle areas in Punjab, Pakistan by using larval reared in large numbers. Unfortunately, ticks packet test. A total of 33 farms were screened in many parts of the world have evolved for this purpose from three different districts, resistance to all pesticides available on the namely Sargodha, Okara and Attock located market, driving the development of new in different geographical areas of Punjab, technologies to control this species. Pakistan. The overall prevalence of resistance was 24.2% of the samples collected from Vaccination against ticks using the protein these farms. The percentage of susceptible Bm86 has been shown to be effective against and tolerant strains from all the studied farms acaricide-resistant ticks. This technique has was 36.5 and 39.3%, respectively. The results been successfully implemented in Puerto of this study revealed that when the RF50 Rico for the control of R. microplus on dairy was considered resistant, 45.5, 16.7 and 10% and beef cattle. Observations from Puerto strains were found to be resistant in Districts Rico indicate a potential interaction between of Sargodha, Okara and Attock, respectively. anti-tick vaccination in conjunction with On the other hand, when the RF99 was used, systemic acaricide use. Controlled animal the percentage of resistant strains was raised studies were completed directly comparing to 100% at all farms. It was concluded that the efficacy of vaccination with and without R. microplus strains collected from different systemic acaricide. Additionally, in vitro farms of Punjab and their surrounding areas feeding of ticks with immunoglobulin-G from have great variability in their cypermethrin vaccinated animals with several combinations resistance and susceptibility level, depending of acaricides was used to screen antigen/ on the particular condition at each farm. acaricide combinations and to confirm results of field tests using animals. The results show that the vaccine had a synergistic interaction OA49.06 Comparison of Ixodes Ricinus with the acaricide. Better and longer control Populations in Adjacent Habitats on a was achieved with the combination than when Pasture-Based Dairy Farm either treatment was applied alone. Mr. Taher Zaid1, Ms. Sorcha Brosnan1, Prof. Olaf Schmidt1, Dr. Jason Newton2, Dr. John OA49.05 Diagnosis of Acaricide Resistance Browne1, Ms. Tiphaine Dubourdieu5, Prof. in Cattle Tick in Three Districts of Punjab, Jeremy Gray1, Mr. Jack O’Connor3, Dr. John Pakistan Mee4, Dr. Annetta Zintl1 1University College Dublin, Ireland, 2NERC Life Dr. Zia ud Din Sindhu1, Mr. Muhammad Usman Sciences Mass Spectrometry Facility, United Naseer1, Talha Zafar1, Rehman Hafeez1, Rao Kingdom, 3MSD, Ireland, 4Moorepark Research Zahid Abbas1, Muhammad Kasib Khan1, Bilal Centre, Ireland, 5Paul Sabatier University, Aslam2, Zafar Iqbal1 France 1Department of Parasitology, University of Agriculture, Pakistan, 2Institute of Pharmacy, The distribution of Ixodes ricinus ticks in the Physiology and Pharmacology, University of environment tends to be highly overdispersed Agriculture, Pakistan with greatly divergent densities in adjacent habitats. In order to (1) investigate the The one host tick, Rhipicephalus (R.) mechanisms that underlie this overdispersion microplus is the most important species, and to (2) assess the animal and public health which causes many obstacles in a profitable risks associated with the different habitats, livestock business of bovines. Chemical we compared tick populations in adjacent

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 181 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

habitats with regard to tick abundance, the Research, India, 4National Botanical Research most recent engorgement host and the Institute, India, 5College of Veterinary and prevalence of tick-borne pathogens (TBPs). Animal Sciences, Pookode, Lakkidi P.O., India Five adjacent sites on a dairy farm in Co Kerry were sampled for the presence of ticks using Due to sanguivorous nature, Rhipicephalus standard blanket dragging methods. From microplus, causes severe economic loss each site, 15 nymphs were analysed for C and to the dairy industries globally. Repeated N stable isotope compositions and 40 were application of different generations screened for the presence of TBDs using of chemical acaricides has led to the Taqman qPCR analysis. development of multi-acaricide resistant tick populations. Along with organophospates, Both nymphs and adult ticks were most synthetic pyrethroids and amidines, the tick abundant in hedges bordering the pasture, population in India is developing resistance to somewhat less common at the edge of the ivermectin. Consequently, tick management path and in the woodland and absent from has become a challenge to sustain the animal the centre of the pasture. Isotope analysis productivity. Phyto-formulation and strategic indicated that the nymphs that quested in use of cross-protective vaccine were targeted the woodland had fed on the broadest range to mitigate the situation. By combining of hosts, while qPCR analysis suggested empirical and modern approaches, a highly that nymphs collected from the hedge that potent, widely available plant having anti separated the woodland from the pasture had tick activity was identified. Chemoprofiling the highest prevalence of TBPs (Anaplasma of the active solvent guided extract resulted phagocytophilum, Babesia (cattle or deer in identification of 22 compounds, of which spp) and Borrelia spp). two potent, safe and stable molecules were selected. Integration of solvent guided While there was some overlap in the feeding extracts and active compounds in delivery guilds of hosts parasitized by nymphs in matrix resulted in the development of one the different habitats, nymphs in the hedge natural and another nano-formulation having bordering the woodland seemed to be chiefly >80% anti-tick activity against experimental feeding on one guild (possibly ruminants) challenge infestations. The formulations are while nymphs in the woodland fed on a working through multiple routes resulted broader range of hosts (probably rodents in damage of cuticle, gut and reproductive and birds in addition to ruminants). The organs with significant up and down regulation results also suggest that ticks that detach of multiple genes involved in physiological from livestock or deer on pasture move into process of the tick species. the hedge, boosting the tick numbers and increasing the prevalence of livestock TBPs in Conserved tick proteins, Subolesin, Calreticulin, ticks in this site. Cathepsin, Ferritin-2 and Tropomyosin of Hyalomma anatolicum and Subolesin of R. microplus were identified using RNAi and OA49.07 Multi-Acaricide Resistant Tick in silico analysis. The identified proteins Population: Problem and Mitigation Strategy are expressing significantly (P<0.001) high level in all the stages of the tick species. The Dr. Srikant Ghosh1, Dr. Rajesh Kumar2, Dr. recombinant version of the proteins conferred Satyanshu Kumar3, Dr. Sharad Srivastava4, 65.4% (Subolesin), 41.3% (Calreticulin), Dr. Sanis Juliet5, Dr. Binod Kumar1, Dr. 30.2% (Cathepsin), 51.2% (Ferritin-2) and Manjunathachar V1, Mr. Sonu Sharma3, Ms. 64% (Tropomyosin) protection against H. Ankita Verma2, Dr. Anil Kumar Sharma1, Dr. anatolicum and 65.4% (Subolesin) against Gaurav Nagar1, Dr. Gajanan Chigure1, Dr. R. microplus experimental challenge Shanmugnath C.1, Dr. Parthasarthi BC1, Dr. infestations. The possibility of development Mukesh Shakya1 of multiantigenic antigen formulation for 1Indian Veterinary Research Institute, India, conferring cross-protection against the most 2Indian Agricultural Research Institute, India, economically important ticks was explored. 3Directorate of Medicinal and Aromatic Plants Simulation of both the strategies in field

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 182 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

situation against multi-acaricide resistant tick EHIA was considered for PCs classification infestations is discussed. as bioactive (coumarin and caffeic acid) and non-bioactive (quercetin and rutin). Phytochemical interactions were subsequently OA50 Alternative treatments for assessed combining bioactive:non-bioactive Parasites in Ruminants II PCs (8:2 ratio), and the nature of their interaction was classified using the fractional July 11, 2019, 8:30 - 10:30 inhibitory concentration index (FICindex). Breakout Room 5, Meeting Rooms KLOP, Level 4 All four combinations had a synergetic interaction against larval exsheathment (FICindex < 0.5), and only coumarin:rutin had OA50.01 Anthelmintic-like Activity of no interaction against egg hatching (FICindex Four Polyphenolic Compounds and Their > 0.5). Quercetin and rutin acted as PCs Interactions Against the Cattle Nematode AH-like activity enhancers, reducing EC50 of Cooperia Punctata bioactive molecules in a range of 43% to 64% and 68% to 83% for EHI and LEI, respectively. Dr. Elke von Son-de Fernex1, Dr. Selene Furthermore, coumarin and caffeic acid Escareño-Díaz1, Dr. Miguel Ángel Alonso-Díaz1, bioactivity against free-living stages of C. Dr. Pedro Mendoza-de Gives2, Dr. Epigmenio punctata makes them suitable candidates Castillo-Gallegos1 as markers for anthelmintic-like activity in 1Centro de Enseñanza Investigación y bioactive forages. Extensión en Ganadería Tropical, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico, OA50.02 Effect of Cry5B From Bacillus 2Centro Nacional de Investigaciones en Thuringiensis on Haemonchus Contortus in Parasitología Veterinaria, INIFAP, México Experimentally Infected Sheep

Polyphenolic compounds (PCs) have been John Sanders1, Dr. Raffi Aroian2, Dr. Gary proposed as one of the most bioactive Ostroff2, Dr. Ambily Abraham2, Dr. Kelly secondary metabolites group occurring Flanagan2, David Gazzola2, Dr. Yan Hu2, Tasia in nature, and have been associated to Kellogg2, Dr. Hanchen Li2, Dr. Anne Zajac1 anthelmintic (Ah)-like activity of plants 1Department of Biomedical Sciences and against cattle nematodes. However, little is Pathobiology, Virginia Tech, United States, known regarding their synergetic/antagonistic 2University of Massachusetts Medical School, interactions. This study assessed the in vitro Program in Molecular Medicine, United States anthelmintic-like activity of commercial PCs: quercetin, caffeic acid, rutin and The worldwide rise of anthelmintic resistance coumarin and their combinations against in trichostrongyle nematodes of ruminants the egg hatching and larval exsheathment of has led to the urgent need for alternative Cooperia punctata; one of the most prevalent methods of control. Previous research has nematodes affecting grazing cattle in tropical shown significant anthelmintic effect in vitro regions. The molecules selected for the in and in vivo of Bacillus thuringiensis (Bt) Cry 5B vitro analysis were previously identified crystal protein on several intestinal nematode in bioactive plants through bio-guided parasites of rodents, pigs, and dogs. fractionation. To estimate mean effective concentrations (EC50) five increasing The effect of Cry5B on Haemonchus concentrations were used for both Egg contortus was examined in vitro using hatching (EHIA) and larval exsheathment larval development and adult motility (LEIA) inhibition assays (0.6 to 9.8 mg / assays. Exposure of H. contortus eggs and mL-1 and 0.15 to 2.4 mg / mL-1, respectively). larvae to Cry 5B significantly reduced the From the four molecules, only rutin did not number of third stage larvae produced. affect egg hatching; while quercetin, showed Additionally, in a motility assay, adult H. no bioactivity against eggs or larvae (P > contortus collected from sheep abomasa 0.05). Best-fit EC50 estimated through the and incubated in medium containing Cry5B

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 183 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

showed significantly reduced motility flagrans (strain CIL1A) against GIN in a compared to control worms. To investigate controlled field trial. The experiment was the effect of the compound in vivo, five carried out under Colombian Andes tropical female and five castrated male lambs aged conditions (Mosquera, Colombia). Four approximately 7-8 months were removed different groups of lambs naturally infected from pasture and confined for the duration with GIN were allocated in separate paddocks of the study. All were dewormed to remove and received daily different fungi formulations existing trichostrongyle infection. Sheep at a dosage of 1 x 106 spores/kg of body were subsequently each administered 10,000 weight for 18 weeks. Control group was not third stage H. contortus. Following patency treated with fungi. Parasitological criteria of infection, sheep were divided into two such as the number of eggs per gram of feces groups with similar mean FEC (determined (EPG), the percentage of larvae recovered by Modified McMaster test, detection limit from coprocultures and some others clinical 25 epg). Treatment group sheep were orally and zootechnical indicators were evaluated administered 200 mL of a suspension of weekly. The results were compared using a genetically engineered non-sporulating repeated measures across time model. The Bacillus producing Cry5B daily for 4 days. estimated mean reduction of EPG of the Control group sheep received 200 mL water. developed formulations compared with the FEC were determined daily for 7 days post control group were 62.7%, 79.2% and 62,3% infection (PI) and thereafter every two days for E3, E16 and CS respectively. There was until necropsy at 14 days PI. After 24 hours, a significant reduction (P < 0.05) between mean FEC of treated animals was reduced treated groups and control group. No by 78% compared to controls and by 94% significant differences were observed between after 72 hours. Of the few eggs produced E3 and E16 formulations. The main nematode by treated animals, only 27% developed genera identified were Haemonchus spp., to the L3 compared to >90% of eggs from Trichostrongylus spp., and Teladorsagia spp. control lambs. These results suggest that (resistant Moxidectin-benzimidazole strains) B. thuringiensis Cry5B protein may show and reductions were demonstrated for each promise as a treatment for H. contortus. of these species. It is considered that the reduction of EPG will lead to the reduction of infective larvae in the grass and would result OA50.03 Control of Gastrointestinal in productivity increases in grazing sheep Nematodes in Wool Sheep Using Novel and reduce the use of anthelmintic chemicals. Formulations Based on a Colombian Strain Therefore, use of these formulations will of the Nematophagous Fungi Duddingtonia provide an alternative option to control GIN Flagrans: Reduction in Eggs per Gram of parasites on pasture. Feces Indicator

Jaime Andres Cubides-Cardenas1, Dr. Diego OA50.03 Association of Phytominerals Cortes-Rojas1, Elizabeth Céspedes-Gutierrez1, with the Serum Trace Elements Profile Prof. Jimmy Jolman Vargas-Duarte2, Dr. and Quantitative Gastrointestinal Parasitic Martha Izabel Gómez-Alvarez1 Burden in Grazing Sheep 1Corporacion Colombiana de Investigacion Agropecuaria - Agrosavia, Bioproducts Dr. Rizwan HM1, Dr. Sajid MS2, Dr. Raaziya Department, Colombia, 2Universidad Nacional Nadeem3, Dr. Abdul Qudoos4, Mr. Haider de Colombia, Instituto de Genética, Colombia Abbas1, Mahvish Maqbool2 1Discipline of Parasitology, KBCMA College of Gastrointestinal nematodes (GIN) can reduce Veterinary and Animal Sciences, Narowal Sub or limit sheep production in Colombia. Campus UVAS Lahore, Pakistan, 2Department Nematophagous fungi are natural enemies of of Parasitology, University of Agriculture, GIN and a new alternative for its biological Pakistan, 3Department of Chemistry, control. The aim of this study was to evaluate University of Agriculture, Pakistan, 4Central the effectiveness of three novel formulations Hi-Tech Laboratory, University of Agriculture, (E3, E16 and CS) based on Duddingtonia Pakistan

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 184 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

The grazing meadows and forages are Liquid chromatography coupled to mass important components of the animal spectrometry/electron spray ionization production. Variety, composition, distribution (LC-MS/ESI) was used to characterize and nutritive value of forages in grazing the polyphenolic profile of the OFIEOH areas depend upon the soil type, season, extract. Fifteen compounds were identified and climate. We studied the role of mineral in the OFIEOH extract. Tri-glycosylated supplementation in animals thru selected methyl quercetin derivatives were the main forages in mitigating the gastrointestinal (GI) compounds identified. In vitro egg hatch parasitic threats in grazing sheep. To this end, (EHT) and larval migration tests (LMT) were level of Zinc (Zn), Copper (Cu), Manganese used in a range of concentrations of OFIEOH: (Mn) and Cobalt (Co) were determined in 12.5 to 100 mg/mL for EHT and 12.5 to 200 grazing forages, respective soils and sera mg/mL for LMT. In addition, the LMT was used of the grazing sheep of Sialkot, Punjab, to test ivermectin (IVM), associated with the Pakistan during spring 2015. The correlation inhibitory concentration of 50% (IC50) for of serum trace elements was done with the OFIEOH. The combination of OFIEOH (12.5 burden of GI parasites in grazing sheep. The to 200 mg/mL) plus the IC50 of IVM was prevalence of GI parasites in grazing sheep also tested. OFIEOH got up to 90% efficacy was 32.81% with eight species of GI parasites in the EHT, and 77% in the LMT, showing identified. A significant (P < 0.05) difference a concentration-dependence inhibitory in the level of selected trace elements was effect. We found a drug-extract antagonistic determined in a total of eight collected neutralizing effect when doses of IVM were forages. The mean serum concentrations of added to OFIEOH (maximum efficacy of Zn and Cu were inversely proportional to 73.78%), while a positive additive effect the mean magnitude of EPG in the grazing was observed when OFIEOH was added to sheep. The present study indicated that the IC50 of IVM (IC50 of 82.79 for OFIEOH trace elements (preferably Zn and Cu) rich alone against an IC50 of 55.08 of OFIEOH forages (Cichorium intybus and Cynodon + IVM). Conclusions: OFIEOH alone may be dactylon) have ability to control or lower the considered as a suitable ecofriendly product burden of GI parasites in grazing animals. This to control gastrointestinal parasites of sheep, mitigation approach may eventually develop offering a more holistic approach to improve the resilience against GI parasitic infections animal farming and welfare. The drug-extract particularly in resource-poor countries. interaction is also a promising therapeutic alternative, reducing the final dose to the host, with an optimum combination effect. OA50.04 Opuntia Ficus-Indica (L.) Mill. Hydroalcoholic Extract Against Nematodes of Sheep OA50.05 In Vitro Anthelmintic Efficacy of Citrullus Colocynthis on Haemonchus Dr. Marcelo Molento1, Dr. Carolina Santos1, Dr. Luciano Campestrini2, MSc Douglas Vieira1, Dr. Tauseef Rehman1, Miss Azra Anwer2, Dr Dr. Izanara Pritsch1, Dr. Fabio Yamassaki2, Dr. Khalid Iqbal2 Selma Zawadzki-Baggio2, Dr. Juliana Maurer2 1Cornell University, United States, 2The Islamia 1Univ Federal do Parana / Parasitic University of Bahawalpur, Pakistan Diseases, Brazil, 2Univ. Federal do Parana / Biochemistry, Brazil The appearance of anthelmintic resistance, drug residues that potentially enter the food Opuntia ficus-indica (L.) Mill. is a xerophylous chain, high costs, and the lack of accessibility plant originated in tropical America. The to anthelmintic drugs in distant rural areas aim of this study was to determine the have motivated investigations into novel efficacy O. ficus-indica hydroalcoholic alternatives such as medicinal plants. The extract (OFIEOH) against gastrointestinal present project was designed to evaluate parasites of sheep. Contents: Initially, the the anthelmintic efficacy of Citrullus (C.) hydroalcoholic extract from cladode peels of colocynthis against Haemonchus. In vitro O. ficus-indica was produced by maceration. anthelmintic effects of aqueous-methanol

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 185 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

and ethyl acetate extracts of the fruit of this, costing a further $222 million. C. colocynthis against Haemonchus were Widespread drug resistance urgently determined through egg hatch and adult necessitates the discovery of new therapies motility assays. The effects of four serial to control these infections. Venoms have dilutions of 25 mg/ml of each extract evolved over millions of years to become compared to levamisole (0.55 mg/ml) and cocktails of selective and potent bioactive oxfendazole (three serial dilutions of 25 g/ molecules, but their potential as sources ml) were studied. In the egg hatch assay, of novel antiparasitic compounds has been means of 83.67% and 80.67% of Haemonchusµ underexplored. We screened over 250 crude eggs failed to hatch at the same 25 mg/ml venoms from a diverse panel of spiders, doses of ethyl acetate and aqueous-methanol scorpions, assassin bugs, caterpillars, marine extracts. The ethyl acetate extract exhibited snails, ants and wasps for anthelmintic activity a slightly higher effectiveness than the against the blood-feeding small ruminant aqueous-methanol extract, but statistically, nematode Haemonchus contortus in a larval there was no significant difference (P=0.138). development assay. At 0.2 mg/ml crude In the adult motility assay, the dose of 25 mg/ venom the hit rate for the screen was 20.6%, ml of ethyl acetate extract paralyzed all the with hits dominated by arthropod venoms, worms within only 4h after the start of the particularly tarantulas and ants. Candidate experiment, and the aqueous-methanolic venoms were characterised using bioassay extract of the plant at the maximum guided fractionation to identify the active concentration tested (25 mg/ml) paralyzed all compounds. Five novel small linear peptides worms by 8h post-exposure (P=0.001). were identified from Amazonian stinging ant Results of the present study strongly suggest venoms. Peptides were synthesised and found that fruit extracts of C. colocynthis may to have low micromolar activity against H. present promising alternatives to synthetic contortus larvae (2 – 30 M). The peptides drugs for treating Haemonchus infections. were also injected into adult Lucila cuprina Further in vivo studies to assess bio-availability sheep blowflies, resultingµ in paralysis within of active ingredients from the plant extracts 1 hour of injection at moderate doses (PD50 and minimum non-lethal concentrations for the = 0.5 – 38 nmol/g) and lethality at high safety of livestock treatment are suggested as doses. The peptides were counter-screened needed future studies. for cytotoxicity, haemolysis and pain in vitro using sensory neurons soaked in calcium- fluorescent dye. One peptide showed well OA50.06 Discovery of Novel Amazonian over 100-fold selectivity for H. contortus over Stinging Ant Venom Peptides with Activity mammalian cell lines and no sensory neuronal Against the Sheep Parasites Haemonchus activation. We subsequently generated Contortus and Lucilia Cuprina mutant peptides to improve potency and selectivity against H. contortus and L. cuprina, Ms. Samantha Nixon1,2, Dr. Akello Agwa1, Dr. but found that these were closely correlated. Samuel Robinson1, Dr. Andrew Walker1, Dr. Overall, these findings indicate that Axel Touchard3, Dr. Christina Schroeder1, Prof. arthropod venoms may be a useful source for Irina Vetter1, Dr. Volker Herzig1, Dr. Andrew C. antiparasitic drug discovery. Kotze2, Prof. Glenn F. King1 1Institute For Molecular Bioscience, The University of Queensland, Australia, 2CSIRO OA50.07 Development and Evaluation of In Agriculture and Food, Australia, 3CNRS, UMR Vitro Anthelmintic Activity of Herbal Extracts Ecologie des forêts de Guyane, France on Benzimidazole Resistant Haemonchus Contortus Parasites remain a serious challenge for veterinary medicine and livestock production Mr. Kazim Ali1, Dr. Muhammad Arfan Zaman1, worldwide. The Australian sheep industry Dr. Syed Ehtisham-ul-Haque1, Dr. Muhammad is severely threatened by gastrointestinal Fiaz Qamar1, Miss Rabia Tamkeen2 nematodes, which cost in excess of $430 1Department of Pathobiology, College of million AUD annually. Blowflies exacerbate Veterinary and Animal Sciences, Sub-campus

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 186 63rd Annual Livestock Insect Workers Conference ORAL SESSIONS

University Of Veterinary And Animal Sciences, Pakistan, 2Department of Basic Sciences, College of Veterinary and Animal Sciences, Sub-campus University Of Veterinary And Animal Sciences, Pakistan

The problem of anthelmintic resistance lead the path to make-in-use the herbal extracts (HE) for the treatment of parasitic diseases. The current study was conducted to evaluate the efficacy of medicinal plants against the benzimidazoles (BZ) resistant strains of Haemonchus contortus in small ruminants. Total adult worms (n=1500) that were morphologically identified as H. contortus were frequently collected from different small ruminant abattoirs covering two geographic regions of Punjab Province (Pakistan). These worms were confirmed using PCR and then subjected for DNA gene sequencing to isolate the resistant strains. The DNA sequences of the resistant strains were deposited into NCBI GENBANK database with Accession No. MF043121-28. Worms were treated in vitro with combined aqueous extract of Nigella sativa and Tachyspermum ammi. Anthelmintic activity of the HE has been evaluated by using the Egg Hatch Test (EHT) and Adult Mortality Assay (AMA). Dose-and- time dependent responses of the HE against the resistant strains were recorded. The results obtained from AMA were statistically evaluated by one-way ANOVA. Mortality of worms was comparable with the reference drug levamisole at the maximum dose rate of the HE (200mg/ml). Data from EHT had been evaluated by Probit model test and lethal concentrations were calculated. HE affects the hatching of eggs in dose dependent manner. The LC50 values of the experiments were distinctly different. The P values for the experiments for efficacy of HE concentrations and oxfendazole were recorded as 0.263 and 0.192, respectively. In conclusion, the HE efficacy is comparable with the efficacy of the synthetic anthelmintics. It can be used being field friendly, environmently safe, socially efficient, cheap and easily available alternatives. Moreover use of HE in treating the parasitic problems will help to reduce the dilemma of anthelmintic resistance.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 187 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 188 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Poster Sessions

Poster Session 01 in 2 out of 3 cats after 14 days of treatment and were still observed in little amounts at July 8, 2019, 10:00 – 16:30 D28 for both cats and at D58 for one cat. Exhibit Hall A, Level 1 Other hematology and blood biochemistry parameters remained unaffected.

PS01.01 Preliminary Safety Study of Miltefosine Miltefosine administered orally at the dose- in Healthy Cats Treated During 14 Days level of 2 mg/kg for 14 consecutive days induced sporadic vomiting in one out of three Dr. Caroline Bouchez1, Dr. Emmanuel Briant1, cats. At hematology, the number of Heinz Dr. Magali Dolon1, Dr. Christelle Navarro1, Dr. bodies increased in two out of three cats Vanessa Chala1 with no consequence on other hematology 1Virbac, France parameters.

Feline leishmaniosis is a chronic disease due to Leishmania infantum with clinical signs PS01.02 Effect of a Long-Acting Moxidectin and laboratory abnormalities similar to those on the Control of Gastrointestinal Nematodes found in dogs. Miltefosine (Milteforan®, Virbac) and the Productivity of Suckling Calves has got veterinary marketing authorizations in Europe and Brazil to control canine Fernando Borges1, Mario Conde1, Mariana leishmaniasis. The objective of this non- Freitas1, Matheus Nakatani1, Vinícius controlled study was to evaluate the clinical Rodrigues1, Guilherme Reckziegel1, Fernando tolerance of miltefosine on healthy cats. Aono2, Elio Moro2, Frederico Moreira3 1Ufms, Campo Grande, Brazil, 2Zoetis Animal After a 7-day acclimation period, three cats Health, Brazil, 3Zoetis Animal Health, United (two males, one female, weighing 3.7 to States 6.1 kg) received Milteforan® once a day on their food ration at the dose-level of 2 mg The impact of gastrointestinal nematodes miltefosine/kg for 14 consecutive days (D0 (GINs) on different categories of beef cattle in to D13) followed by a 14-day recovery period. tropical production systems requires studies Animals were observed daily from D-7 to and little is known about the advantages of D28. Product consumption was recorded 30 anthelmintic treatment in pre-weaned calves. minutes to 1 hour after each treatment and The objective of this study was to evaluate the food consumption daily. Clinical follow-up was effect of a long-acting formulation containing performed before each treatment and clinical 10% moxidectin on the control of GINs and examination including rectal temperature, the productivity of calves before weaning. A body weight measurement performed weekly. first study was designed to evaluate the effect Blood samples were taken at D-6, D14, D28 for of the treatment of calves between 3 and 5 hematology and blood biochemistry, at D58 months of age, and later ten other clinical for hematology. studies were carried out in different Brazilian states to evaluate the repeatability of these All animals ingested spontaneously the results. In the first study, 202 animals were expected dose-volume of product for 14 distributed in two groups (moxidectin 500 g/ consecutive days, showing good palatability. kg or Saline) in a randomized block design, One out of three cats vomited 4 times during considering the weight, fecal egg counts μ the treatment period, without impact on (FECs), breed, and dam’s parity. Other ten health status. No other clinical signs possibly field studies were carried out in commercial related to treatment was reported. Rectal farms from four Brazilian geographic regions, temperature, body weight, food consumption where 692 calves were randomly assigned remained unaffected. Heinz bodies increased to receive either 10% moxidectin or saline

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 189 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

treatments. Calves treated with moxidectin This tool has been validated according to the in the first study had an increase of 9.4 recommendations of the French standard kg on the body weight (BW) (P<0.05), a NFU47-600-2. reduction in mean OPG (P<0.05) and a higher percentage of negative FECs (56.8% vs. 36%; Thus, for the PCR part, the following P<0.05). In the other 10 field studies, calves characteristics were determined: LODPCR (5 treated with moxidectin had an increase of copies / PCR), Efficiency (99%) and Linearity 4.8 kg (p < 0.02) and a greater average daily Domain (between 10 copies corresponding to BWG (0.704 vs. 0.653 kg/d; p<0.001). The the LOQPCR and 1.106 copies / PCR). FECs decreased 68.7% from the treatment to the weaning in the treated calves, whereas in Various modes of faecal preparation were the placebo group, there was an increase of compared in terms of the extraction yield 23.2% in this interval. Therefore, the treatment of known faeces positives for Giardia: either of suckling calves with long acting moxidectin mechanical lysis, or thermal and chemical reduced FEC and increased weaning weight. lysis, or mechanical lysis followed by thermal and chemical lysis; or no pretreatment at all before nucleic acids extraction-purification. It PS01.03 Validation of a Semi-Quantitative is the association of mechanical and thermal- Real-Time PCR Assay for the Diagnosis and chemical lysis that reproducibly gives the best the Monitoring of Giardia Intestinalis Infection extraction yield. in Canine Faeces Samples According to the French Standard NFU47-600-2 Finally, the complete method part was characterized by the determination of the Florence Va1, Myriam Thomas2, Sylvain diagnostic sensitivity and specificity. We Bellier1,3, Claire Ciancia4, Valeriia Klubkova4, analyzed a hundred dog faeces with or without Claire Pelletier5, Eric Sellal4,5, Bruno Polack1,3 an evocative clinic in order to determine an 1BioPôle Alfort, Ecole Nationale Vétérinaire infection threshold of clinical interest making d’Alfort, France, 2UMR BIPAR, Anses, Ecole it possible to distinguish between a chronic Nationale Vétérinaire d’Alfort, INRA, Université carriage and a clinical infection. Paris-Est, Animal Health Laboratory, France, 3UMR BIPAR, Ecole Nationale Vétérinaire Because of its wide range of inclusivity (7 d’Alfort, ANSES, INRA, Université Paris-Est, recognized assemblies and exogenous IPC), France, 4BioSellal, France, 5BioDev, France the qPCR Premium® Giardia intestinalis kit can also be used on other animal species such as Infection of young dogs with Giardia cats, ruminants or even environmental samples. intestinalis is very common in France and is a major clinical problem, especially in breeding facilities. PS01.04 Development of a Diagnostic Assay for the Detection and Differentiation of In order to obtain a reliable, sensitive and Theileria Spp. In White-Tailed Deer semi-quantitative diagnosis that can easily be carried out in a routine laboratory, we have Mr. Alec Thompson1, Dr. Michael Yabsley1,2 developed a complete tool combining firstly 1Southeastern Cooperative Wildlife Disease a ready-to-use quantitative duplex real-time Study, Department of Population Health, PCR kit (qPCR Premium® Giardia intestinalis College of Veterinary Medicine, University of ; BioDev) to detect all Giardia intestinalis Georgia, United States, 2Warnell School of assemblies A to F (beta-giardin target) by Forestry and Natural Resources, University of FAM labelling and an exogenous internal Georgia, United States positive control (IPC) by Cy5 labelling, and secondly a method of preparation samples The spread of non-native parasites with the and extraction-purification of nucleic acids to movement of animals is a major concern for obtain an optimized yield (BioPrep® Giardia disease emergence and native species health/ and BioExtract® column, BioSellal). conservation. Haemaphysalis longicornis (Asian longhorned tick) is native to eastern

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 190 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Asia, but has become invasive in several distribution, common in freshwater fish. The countries including Japan, Australia, New life cycle takes place between 15-21°C in Zealand, and now the United States. Within 3-4 weeks. In Mexico there are 7 endemic the established range, H. longicornis is a species of axolotl. However, the presence vector of the protozoan parasite, Theileria of L. cyprinacea has only been reported in orientalis subtype Ikeda, which until recently Ambystoma mexicano. In the Atlangatepec was not known to occur in the United States. Aquaculture Center, in the state of Tlaxcala, In 2017, clinical theileriosis caused by T. Mexico, European carps (Cyprinus carpio) orientalis Ikeda was reported in a cattle are raised in ponds for commercial purposes. herd in Virginia which was also infested A group of 8 axolotls (Ambystoma velasci) with H. longicornis, but it is not known if H. also lived in the aquaculture center and were longicornis transmitted T. orientalis to the housed as reproductive couples in separate cattle. Within the United States, white-tailed tanks. The axolotls went to an exhibition deer (WTD, Odocoileus virginianus) are outside the center for two months and their infected with several genotypes of a Theileria tanks were filled with water from the carp’s sp. (often called T. cervi) that is distinct ponds, a species in which Lernaea cyprinacea from T. orientalis. It is currently unknown is commonly found. The temperature of the if deer are susceptible to T. orientalis, a water in the fish tanks housing the axolotls pathogen of agricultural concern. In this had a range of 11-15 ºC. The parasites were study, we developed a restriction fragment collected of all axolotls with the help of length polymorphism (RFLP) assay that can tweezers and placed in plastic tubes with 70% distinguish between Babesia spp., selected alcohol. A total of 22 individual parasites were exotic Theileria spp., and the WTD Theileria obtained and identified at the Parasitology sp. (although not the different genotypes Laboratory, UNAM. The frequency of the [F, G1, and G2]). Using this assay, we tested location of the parasites was: 40.90% (9/22) cervid blood samples from the eastern United in the gills, 18.18% (4/22) in the base of the States and found 222 of 264 (84%) positive neck, 13.63% (3/22) in the groin, 9.09% for the WTD Theileria. sp., 23 (9%) positive for (2/22) in the eyes, 4.54% (1/22) in the right Babesia spp., and 2 (<1%) for exotic Theileria shoulder and 4.54% (1/22) in the base of the spp. Seven of the 264 samples had insufficient cloacal glands. In addition, fibrous nodules quantities of DNA to visualize RFLP results. were observed in axolotls where Lernaea Sequencing of 20 selected samples confirmed cyprinacea was removed from the skin and RFLP results. Sequences of the two ‘exotic’ gills. It was concluded that the water from the Theileria amplicons were >99% similar to T. ponds where the carps were raised was the ovis, a nonpathogenic ovine parasite native source of contamination for the axolotls. In to Asia. Our data indicate that WTD Theileria. addition, it was speculated that the parasites sp. infections are common and that WTD adapted well to the low temperature of the can possibly maintain a higher diversity of axolotls’ tanks and thus, they survived. Theileria spp. than recognized.

PS01.06 Susceptibility of Selected Strains of PS01.05 Report of Lernaea Cyprinacea in Cat Fleas, House Flies and Darkling Beetles Axolotl Serrano (Ambystoma Velasci) in the Obtained From Selected Laboratory Colonies Municipality of Atlangatepec in Tlaxcala, to Deltamethrin, Fipronil and Imidacloprid Mexico Bill Donahue1 Prof. Maria Cristina Guerrero-Molina1, Prof. 1Sierra Research Laboratories, Inc., United Ángel García-Hernández1, Prof. José Manuel States Cobo-González1, Lidia Boleaga-Rivera1, Itzel Hernández-Aranda1, Natalia Vieyra-Gómez1 Veterinary pests such as cat fleas, house 1Facultad De Medicina Veterinaria Y Zootecnia- flies, and litter beetles are routinely reared in UNAM, Cd Universitaria CDMX, Mexico entomology laboratories around the world. Bioassays are conducted in the laboratory to Lernaea cyprinacea is a parasite of worldwide determine the efficacy of new compounds

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 191 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

and formulations which may be used for location of adult worms within dogs, cats, and product registration or to conduct product multimammate rats. In the present study, an comparisons for marketing claims. Concerns attempt was made to restrict the location of regarding insect health, vigor, behavior adult Brugia spp. within Mongolian gerbils. and genetic diversity should be addressed Two groups of 5 male gerbils each were and characterized to better understand the infected subcutaneously in the dorsal neck results obtained during testing. Whereas with either 75 infective third-stage larvae of standardization of colonies may be B. malayi or B.pahangi. The animals were impractical and in many cases undesireable, maintained for 90-92 days before necropsy. characterization of colonies should be Just prior to necropsy, blood was collected documented. from each animal for determination of patency. A complete necropsy was done on For example, insecticide resistance from each animal with examination of lymphatics, field populations can be quite variable within testes, hearts and lungs. All dissected tissues laboratory reared colonies, often being lost and carcasses were soaked overnight and if not periodically challenged or infused with the soakings were examined for worms. Four new field collected stock. Baseline data from gerbils infected with B. malayi had worms. new insect collections should be established Recoveries ranged from 2-12 worms (5.5 as soon as colonies are reproductively viable worms per animal, average) with all but 1 and evaluated periodically to note changes female worm being recovered from the heart in susceptibility to selected classes of and lungs. This single female worm was insecticides. recovered from the left testicle of the gerbil that had 12 worms. All 5 gerbils infected with Results from dose response bioassays B. pahangi had worms and all of the worms will be presented for several strains of were recovered from the heart and lungs. cat fleas, house flies and litter beetles Recoveries of B. pahangi ranged from 1-19 from selected laboratory colonies used (5.4 worms per animal, average). Microfilariae for product development and regulatory were found circulating in the blood of only 1 activities. Technical deltamethrin, fipronil gerbil infected with B. malayi. This gerbil had and imidacloprid were serially diluted in 12 worms (10 females, 2 males). None of the acetone and applied to various substrates. animals infected with B. pahangi were patent. Adult insects were confined to the treated This initial success at limiting the location of substrates and continuously confined to the worms to the heart and lungs of gerbils may treated surface. Knockdown and mortality lead to the validation of a rodent model for were assessed at predetermined time intervals canine heartworm infection. through 48 hours. Probit analysis (Polo Plus ver 2.0) was used to determine lethal dose values at selected times after exposure to the PS01.08 Effects of Protein Diet Level and treated substrate. Haemonchus Contortus Infection on Lamb Performance

PS01.07 Preliminary Study to Localize Adult Camila Chaves1, Paula Henriques1, Victor Brugia Malayi and Brugia Pahangi Within Nakaguchi1, Isabela Moraes1, Alessandro Mongolian Gerbils (Meriones Unguiculatus) Amarante2, Luciana Katiki3, Adibe Abdalla1, Following Subcutaneous Injection of Helder Louvandini1 Infective Third-Stage Larvae 1Centro de Energia Nuclear na Agricultura, Brazil, 2Universidade Estadual Paulista, Brazil, Michael Dzimianski1, Erica Burkman1, 3Instituto de Zootecnia, Brazil Christopher Evans1, Andrew Moorhead1 1The University of Georgia, United States The aim of this study was to verify the effects of diet protein content and Haemonchus The site of subcutaneous infection with contortus infection in sheep performance. infective stage larvae of Brugia malayi and Thirty Santa Inês lambs (16 females and 14 B. pahangi has been utilized to limit the castrated males), six months old; initial body

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 192 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

weight (BW) 24.9 ± 4.03 kg (mean ± SD) were their responses to experimental infection. randomly distributed in a factorial arrangment To analyze the effectiveness of this strategy 2x2, with high (HP: 17%) and low (LP: 10%) and its potential effects on growth traits, we protein levels, and infected (i) and non- divergently selected sheep of the Romane infected (c, control) with H. contortus, coming meat breed based on their response to up to four treatments: HPc (n = 6), HPi (n = Haemonchus contortus experimental 10), LPc (n = 6) and LPi (n = 10). On infected infections. A total of 91 naïve female lambs of groups each lamb received a single dose of the second generation (51 from the resistant 10,000 H. contortus L3 larvae. Fortnightly, the line and 40 from the susceptible line) were animals were weighted (BW). During 62 days, successively challenged with 3,500 larvae and daily dry matter intake (DMI) was recorded 10,000 larvae of H. contortus at 3 to 5 months for feed intake (FI) determination in g DMI/ of age. In all lambs, we measured fecal egg kg BW0.75. No differences were observed count (FEC), hematocrit, weight, back fat on worm burdens between infected groups and muscle thickness at the beginning and (P>0.05). The male lambs (30.1 kg) were at the end of the second infection, and twice heavier than females (26.5 kg) (P<0.05), but a week during the second infection for the there was no interaction between sex and 42 most divergent lambs. Lambs from the treatments (P>0.05). There was protein and S line excreted on average ten times more infection interaction on FI and BW (P<0.05). eggs than those from the R line during the The HPi group (66.3 g DMI//kg BW0.75) had second infection. Although growth in body a lower FI (P<0.0001) than HPc (69.1 g DMI/ mass did not differ between lines, growth in kg BW0.75), LPc (69.3 g DMI/kg BW0.75) fat thickness was 20% higher in the S line than and LPi (69.5 g DMI/kg BW0.75). Lambs from in the R line. Overall, selection for resistance HPc presented a significant (P<0.001) higher to H. contortus based on experimental BW (30.0 kg) in comparison to HPi (26.8 kg), infestation apparently resulted in highly while the other groups showed intermediate effective immune response in growing female values LPc (28.4 kg) and LPi (28.3 kg). In lambs but may compromise fat deposition, conclusion growing lambs when fed with suggesting a potential energetic cost of higher protein presented better performance, immunity against GIN infection. but they were more affected on FI and BW by H. contortus infection, than those with lower protein availability in the diet. PS01.10 Resistance of Helminths to Closantel 10%, Albendazole 5% and Praziquantel 7.5% in Hampshire Down Sheep from the PS01.09 Highly Effective Selection for Department of Misiones, Paraguay Resistance to Haemonchus Contortus Experimental Infections May Lead to Dr. José Mariano Riveros1, Dr. Antonio Reduced Fat Deposition in Growing Sheep Rodríguez2, Dr. Jorge Miret1 1Facultad de Ciencias Agropecuarias y Sophie Aguerre1, Dr. Frederic Douhard1, Dr. Ambientales (FCAA) Universidad Nacional de Christelle Griesez2, Francoise Prevot2, Didier Canindeyú (UNICAN), Paraguay, 2Centro de Marcon3, Dr. Jean-Louis Weisbecker1, Dr. Diagnóstico Veterinario (CEDIVEP), Paraguay Carole Moreno1, Prof. Philippe Jacquiet2 1GenPhySE, Université de Toulouse, INPT, Gastrointestinal nematodes is a serious INPT-ENVT, France, 2INRA-ENVT UMR1225, problem in sheep production, causing a France, 3Domaine expérimental Bourges-La decrease in body condition, weight loss, Sapinière INRA, UE 0332, France reduction of productivity, and in severe cases death of the animal. The objective of As anthelmintic resistance is now widespread this research was to evaluate the efficacy in gastro-intestinal nematodes (GIN), genetic of closantel 10%, albendazole 5% (Microtel, selection of sheep for a better resistance to Laboratorio Microsules, Uruguay. S. A) and GIN infection is increasingly considered as praziquantel 7.5% (Oviquantel, Vetanco, an alternative strategy. One possibility for Argentina) in 32, 7 months to 6 years this is to select breeding animals based on Hampshire down sheep from both sex from a

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 193 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

farm of the Department of Misiones, Paraguay. of the body of Harris’Hawk (Parabuteo The anthelmintic resistance was determined unicinctus) young birds from Morelos, in by McMaster technique (described by Coles et Mexico. Sixty-eight fly larvae of forunculus al., 1992), before and 14 days after treatment. lesions were collected from the head, chest The helminthes species identification was region, thighs, legs and phalanges of two one- carried out by coproculture and identification month-old Harris’Hawk (Parabuteo unicinctus), of stage 3 larvae (L3), (Fiel et al., 2011). High from the state of Morelos in Mexico. In the resistance to anthelmintic treatment was Parasitology Laboratory of the Faculty of observed since only 6/32 sheep showed Veterinary Medicine and Animal Husbandry of reduction in eggs counts ≥ 90%. Sheep with the UNAM, the larvae were placed between less than 1 year showed at the coproculture the holder and covered objects with Hoyer’s and identification of L3: Ostertagia sp 6%, liquid. The posterior respiratory stigmas of the Cooperia sp 38%, Haemonchus sp 43% and larvae were observed under a stereoscopic Trichostrongylus 5%. Adult sheep: Ostertagia microscope. After observing the morphological sp 6%, Cooperia sp 19%, Haemonchus sp characteristics of the larvae, it was determined 67% and Nematodirus sp 8%. This research that they were larvae of the genus Philornis sp. shows the high resistance to helminths in The importance of this communication is that sheep from the Department of Misiones to the injuries caused by the larvae of Philornis sp, the three molecules tested and that should be in the muscular and cartilaginous tissues can implemented prophylactic measures that will produce irreversible deformations that affect reduce the problem along the production such the movement of the birds. as feeding supplementary, pasture rotation and adequate use of anthelmintics in sheep. PS01.12 A Retrospective Study on the Occurrence of Parasitic Diseases of Wild PS01.11 Infestation with Larvae of the Fly Animals in Western Ghats of Karnataka, India Philornis Sp. Of Harris’Hawk (Parabuteo Unicincus) Young Birds in Morelos, Mexico Ananda Konanur Javaregowda1, B. Kavitha Rani1, S. S. Manjunath1, S. Vinay1 Prof. Maria Cristina Guerrero-Molina1, Prof. 1Department Of Veterinary Parasitology, Cintli Martínez Ortiz de Montellano1, Prof. Karnataka Veterinary, Animal and Fisheries Yazmin Alcala-Canto1, Prof. Juan Carlos Sciences University (b), Veterinary C, India Morales Luna1 1Facultad De Medicina Veterinaria Y Wildlife is our national treasure and the wild Zootecnia-UNAM, Cd Universitaria Cdmx, animals are susceptible to wide variety of Mexico parasitic diseases which often go unnoticed. Among factors that threaten the existence The fly of the genus Philornis (Diptera, of wild animals, infectious diseases in Muscidae) has 51 species and a distribution is particular those arising from parasites play mainly Neotropical. Their larvae are parasites an important role. The present study reports of only subcutaneous location and feed the occurrence of parasitic diseases in on blood from a wide range of birds. The wild animals, both captive and free range development of the larva is rapid between received for necropsy in Western Ghats 4 and 6 days and forms boils that extend of south-central Karnataka from 2009 to through the dermal openings of its hosts. The 2018. The necropsy of tigers showed Taenia black-and-white Hawk (Parabuteo unicincus) sp., Ancylostoma sp., Paragonimus sp. and is distributed from the southwestern United Toxocara sp. Leopards harboured Spirometra States of North America, in Mexico, central sp., Taenia sp. and Physaloptera sp. The Argentina and southern Chile. This bird is necropsy of elephants showed Amphistome used frequently in falconry, which is why sp., Cubboldia elephantis bots and Murshidia it is sometimes raised and marketed in a sp. of strongyles. The necropsy of Pythons particular way. The objective of this work was showed infections with Ophidascaris sp. and to communicate the presence of larvae of flies Ascaridia galli whereas; Checkered Keelback of the genus Philornis sp, in different regions snake had Ascaridia galli. The post-mortem

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 194 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

examination of peacock showed Ascaridia inhibitor of thrombin, blocking its coagulation galli and Heterachis gallinarum species, Ibis of plasma and inhibiting S-2238 hydrolysis in a bird had Echinostoma sp. and Ascaridia competitive manner with an inhibition constant galli whereas, Red Jungle fowls showed (Ki) of 7.7 pM. Using isothermal titration Heterachis gallinarum in the caecum. The calorimetry, we observed XC-43 and thrombin intestinal contents collected during necropsy binding reaction reaction was enthalpy-driven were examined by qualitative methods for ( H = −2.95 x 104 kcal/mol) with an equilibrium detection of parasitic ova. The intestinal constant (K) of 1.33 × 10-10 M−1. Although contents of tiger showed the eggs of above XC-43Δ has a putative thrombin cleavage said parasites along with Isosporan oocyst. site (Lys11-Met12), peptide is not cleaved by Similarly, in species like leopard, python, thrombin. Evaluation of XC-43 anticoagulant Checkered Keelback and Ibis, the eggs properties in vivo is underway. The identification observed corresponded to the parasites of novel natural anticoagulants and the mentioned above respectively. In elephants, understanding of their mechanism of action along with ova of the above said parasites, may offer opportunities for designing new there were also eggs of Fasciola sp. and antithrombotics disrupting blood clotting. Schistosoma sp. Whereas, in Peacock and Taking into account the selectivity for thrombin Red Jungle fowls, the ova of Ascaridia galli and its kinetic properties, this peptide could be and Heterachis gallinarum were observed a candidate for clinical use as an antithrombotic. along with coccidian oocysts. The present study reports the existence of wide variety of parasitic diseases among wild animals PS01.14 Mosquito Distribution and Duck which is infectious with a capacity to spread, Tembusu Virus Infection on Duck Farms in probably also with zoonotic potential. Sing Buri and Ang Thong Provinces, Central Thailand

PS01.13 Functional Characterization of a Jitra Sanisuriwong1, Nichapat Yurayart1, Dr. Salivary Thrombin Inhibitor from the Flea Aunyaratana Thontiravong2, Dr. Sonthaya Xenopsylla Cheopis Tiawsirisup1 1Animal Vector-Borne Disease Research Unit, Dr. Lucas Tirloni1, Dr. Jose M. C. Ribeiro1, Dr. Veterinary Parasitology Unit, Department of John F. Andersen1 Veterinary Pathology, Faculty of Veterinary 1Laboratory of Malaria and Vector Research, Science, Chulalongkorn University, Thailand, National Institute of Allergy and Infectious 2Department of Veterinary Microbiology, Diseases, National Institutes of Health, United Faculty of Veterinary Science, Chulalongkorn States University, Thailand

The salivary glands of hematophagous animals Duck Tembusu virus (DTMUV) is an emerging contain a complex cocktail that interferes infectious disease in ducks, belonging to the with the host hemostasis and inflammation Ntaya virus group of the Flavivirus genus and pathways, thus increasing feeding success. Flaviviridae family. The emergence of DTMUV Analysis of the salivary gland transcriptome has been observed on layer and breeder of the flea Xenopsylla cheopis, the vector of duck farms in China since 2010 as well as human plague, reveals presence of a 59-aa on layer and broiler duck farms in Thailand long peptide (named XC-43) containing a since 2013. Infected ducks show neurologic signal peptide, and without significant matches signs, including an incapability to stand, to other proteins in public databases. Mature ataxia, and paralysis. A significant drop in XC-43 (36-aa long) was synthesized and used egg production is usually observed among in coagulation assays with human plasma, layer ducks. The transmission of DTUMV where we were able to observe anticoagulant involves insect vectors, with mosquitoes properties, delaying thrombin time (TT), also being important vectors for this virus. prothrombin time (PT), and partially activated However, the exact role of mosquitoes in thromboplastin time (aPTT). XC-43 was the ecology of DTMUV in Thailand remains found to be a specific, fast and tight-binding unclear. This study was conducted to examine

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 195 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

mosquito distribution and DTMUV infection in of Anycylstoma caninum ACR-16 as a mosquitoes on duck farms in central Thailand. drug target using two-electrode voltage- Mosquitoes were collected from two duck clamp electrophysiology. This technique farms in Sing Buri Province and two duck allowed us to explore the effects of several farms in Ang Thong Province from September cholinergic agonists and antagonists on 2015 to July 2016. Four CDC-light traps were the nAChRs expressed in Xenopus laevis used for collecting mosquitoes from each oocytes. Acn-ACR-16 was not sensitive duck farm and were operated overnight. A to many of the existing cholinomimetic total of 30,841 mosquitoes were collected anthelmintics (levamisole, oxantel, pyrantel, and identified. Seven mosquito species were morantel, bephenium and tribendimidine). found in this study, comprising Anopheles 3-Bromocytisine was the most potent barbirostris, An. stephensi, Culex gelidus, agonist (>130% of the control acetylcholine Cx. quinquefasciatus, Cx. tritaeniorhynchus, current). Unlike Asu-ACR-16, the Anycylstoma Mansonia annulifera, and Ma. uniformis. caninum nAChR did not produce a response The most collected mosquitoes from each to oxantel. The mean time constants of duck farm and each collected time were Cx. agonists for desensitization rates ranged tritaeniorhynchus. The pools of mosquitoes between 1.5 and 4.8s for Acn-ACR-16 and were then examined for DTMUV infection by were longer than the rates observed in Asu- RT-PCR using specific primers. A total of 273 ACR-16. In contrast to Asu-ACR-16, the A. mosquito pools were examined, with only one caninum receptor was completely inhibited by pool of Cx. tritaeniorhynchus collected from Dh E and moderately inhibited by -BTX. In Sing Buri Province in November 2015 testing conclusion, we have successfully reconstituted positive for DTMUV. Thus, this study indicates a fullyβ functional homomeric nAChR,α ACR- that Cx. tritaeniorhynchus may play an 16 from A. caninum, which is used as a important role as a vector in the transmission model for human hookworm infections. The of DTMUV in Thailand. However, additional pharmacology of the receptor is distinct from studies concerning the vector competence of the levamisole sensitive nematode receptors. this mosquito for DTMUV are needed. The ACR-16 homologue also displayed some pharmacological differences from Asu- ACR-16. Hence, A. caninum ACR-16 may be PS01.15 Pharmacological Characterization a valid target site for the development of of a Homomeric Nicotinic Acetylcholine agents against hookworm infection. Receptor Formed by Ancylostoma Caninum ACR-16 PS01.16 Anthelmintic Effect of Shivani Choudhary1, James G. Tipton1, Supplementation with Mimosa Melanie Abongwa1, Matthew Brewer1, Jeba Caesalpiniifolia and Feeding Behavior of J. Chelladurai1, Nicole Musselman1, Richard J. Goats Grazing in Tropical Deciduous Forest Martin1, Alan P. Robertson1 1Iowa State University, United States Nathalie Silva Pinto1, Danilo Rodrigues Barros Brito2, Ellen Silva1, José Cutrim-Junior2, Pedro Parasitic infections are treated using Gozalez-Pech3, Felipe Torres-Acosta3, Helder anthelmintic drugs, some of which target Louvandini4, Carolina Rocha Silva1, Naylene nicotinic acetylcholine receptors located Carvalho Sales Silva1, Livio Costa-junior1 in different tissues. The limited arsenal of 1Federal University of Maranhão, Brazil, anthelmintic agents and the prevalence of 2Federal institute of education science and drug resistance implies that future defense technology of Maranhão, Brazil, 3Universidad against parasitic infections will depend Autónoma de Yucatán, Mexico, 4State on the discovery of novel targets and University of São Paulo, Brazil therapeutics. Previous studies have shown Ascaris suum ACR-16 nicotinic acetylcholine The use of tropical deciduous forests receptors to be a suitable target for the (TDF) is an important resource in goat development of antinematodal drugs. In this production, due to the great diversity of study we characterized the pharmacology fodder plants rich in protein and condensed

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 196 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

tannins (CT), which may be important in the Jesse Pawlak1, Erica Ward1, Amanda Kuzyk1, control of gastrointestinal nematodes. The Haley Silas1, Dr. John Gilleard1 objective of the study was to evaluate the 1University Of Calgary, Canada, 2University of effect of supplementation of an important Saskatchewan, Canada tanninipherous plant, Mimosa caesalpiniifolia, Gastrointestinal nematodes are a major against H.contortus in goats grazing in problem for sheep industry worldwide. TDF. Egg hatching, larval exsheathment, Nematode species differ in their clinical bromatological analysis and chromatographic manifestations, economic impacts, profile of the sample plant, were performed. epidemiology and drug sensitivities. Twenty-four goats were infected with a single Consequently, quantifying the species dose of infective larvae of H.contortus, 28 days composition of parasite communities is prior to the start the experiment. The animals important to direct drug treatment choice, were distributed into two groups according diagnose drug resistance and inform parasite to fecal eggs count and body weight. Group management strategies. The recently I receiving concentrate with M.caesalpiniifolia developed next-generation deep amplicon (128.7 mg/CT/kg) and Group II:receiving ITS-2 rDNA nemabiome sequencing, concentrate without M.caesalpiniifolia. All using the Illumina Miseq platform, has animals received isoproteic and isoenergetic many applications in this area due to its supplementation and have daily access to high accuracy, throughput, sensitivity and TDF. Animals were weighed weekly and faecal increasing affordability. Here we validate egg counts were performed twice per week. deep amplicon ITS-2 rDNA sequencing for The direct observation method was applied ovine gastrointestinal nematodes and show in 4 goats to record the species of plants its reliability in the relative quantification of consumed. After 28 days of experimental nematode species from either eggs, L1 or feeding, all animals were humanly slaughtered L3 larvae harvested from fecal samples. We and adult worm populations were estimated. have applied the validated assay to a large A total of 20 species of TDF plants were scale study of more than 40 farms across consumed. The medium values of dry matter Western Canada over 5 years to investigate and crude protein Intake was significantly parasite species distribution and anthelmintic different between the groups(P<0.05). The resistance. Nemabiome sequencing was in vitro exsheathment inhibitory effect (IC50) applied to samples collected by producers was 0.46mg/mL. The low inhibitory effect before and after drug treatment and mailed was observed in egg hatch test. The reduction to the laboratory. We also visited farms in of faecal egg counts was 25%. There was no Alberta and performed Fecal Egg Count significant effect of the supplementation with Reduction tests over several years to M.caesalpiniifolia on the reduction of adult investigate benzimidazole, ivermectin and worm burdens. There is significant potential in closantel resistance. Nemabiome sequencing the use of M.caesalpiniifolia in the goat diet for was used to help confirm resistance and the control of H.contortus, but there is a need determine which parasite species were for other studies to clarify in more detail how resistant to which drug. Haemonchus the anthelmintic effect of the tannins or other contortus was the most prevalent species polyphenols present in the plant studied. on most farms and was most commonly found to be resistant to both ivermectin and benzimidazoles followed by Teladorsagia PS01.17 The Use of Deep Amplicon ITS-2 circumcincta and Trichostrongylus rDNA Nemabiome Sequencing to Investigate colubriformis. Nemabiome sequencing was Anthelmintic Resistance in Ovine Gastro- also instrumental in determining that the low Intestinal Parasites efficacy of closantel in many Fecal Egg Count Reduction tests was not due to anthelmintic Dr. Camila Queiroz1, Dr. Russell Avramenko1, resistance but due to the narrow spectrum Dr. Elisabeth Redman1, Dr. Fabienne of the drug. Results show that ITS-2 rDNA Uehlinger2, Dr. Michel Levy1, Dr. Lana Swain1, nemabiome sequencing is a valuable adjunct Dr. Kelsey Kearns1, Dr. Amelia Whitelaw1, to the FECRT in investigating anthelmintic Elspeth Yates1, Erika Brandson1, Jeff Lees1, resistance.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 197 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS01.18 Exploring Thermosensory Behaviours foresee this approach enabling the continued and Pathways in Filarial Nematode Parasites identification and annotation of D. immitis thermosensory effectors, which could Mr. Zachary Heimark1, Mrs. Katherine Kudrna1, be putative targets for the prevention of Dr. Nicolas Wheeler1, Dr. Mostafa Zamanian1 heartworm infections. 1University Of Wisconsin-Madison, United States

Dirofilaria immitis is the etiologic agent PS01.19 Study of the Role of of canine and feline heartworm disease. P-Glycoprotein-2 of the Parasitic Nematode The prevention of this disease is achieved Heligmosomoides Polygyrus in the through the administration of macrocyclic Cholesterol Uptake lactones as a prophylactic, or melarsomine dihydrochloride in dogs with adult infections. Mohamed Issouf1, Fatima Mohamed1, Cédric Severely limited pharmacologic options make Neveu1 control of this disease difficult, especially 1INRA, UR1282 Infectiologie Animale et Santé in regions of drug resistance or areas Publique, Centre de Recherches de Tours, with high non-compliance of preventative France supplementation. Therefore, in order to decrease the reliance on a single-drug Recent studies have described the group, other prevention strategies need involvement of Caenorhabditis elegans to be explored. L3 and L4 larval stages p-glycoprotein-2 in lysosome formation and of D. immitis are the targets for existing lipid storage. In that respect, we are carefully chemotherapies, and these stages exhibit trying to identify in Heligmosomoides thermosensory behaviours in vitro, which are polygyrus bakeri the free-living nematode likely used during transmission and infection model Caenorhabtidis elegans p-glycoprotein of the mammalian host. This behaviour homologs using degenerate primers. These could be exploited by chemotherapeutic experiments have allowed the identification of intervention, but the molecular effectors of five partial p-glycoprotein sequences. Among thermosensation are unknown in D. immitis. those Hba-pgp-2 has particular interest. To identify putative thermosensory effects Since, C. elegans PGP-2 is involved in the in D. immitis, we conducted a pan-phylum transport of cholesterol. Cholesterol is needed comparative genomic and phylogenetic for molting and growth of the nematodes. analysis informed by thermosensory In this study, a full-length cDNA sequence pathways elucidated in the model nematode corresponding to Hba-pgp-2 was obtained. Caenorhabditis elegans. To validate these It presents high homologies with the Cel- putative thermosensory effectors, we pgp-2 gene sequence. Moreover, we showed developed a reverse genetics approach to that Hba-pgp-2 is expressed among the knock-down expression of gene targets eggs, fourth-stage larvae and adult and their predicted to function in thermosensation and expression could be induced by cholesterol. In assayed the modulation of thermosensory order to characterize the Hba-pgp-2 function, phenotypes. Because of the fragmented the Hba-pgp-2 promoter was analyzed nature of D. immitis gene predictions, we using the nematode model C. elegans and used long-read isoform sequencing of complementation of C. elegans pgp-2 mutant adult males and females to annotate full- by Hba-pgp-2 was performed. This opens the length genes, which enabled the design of way for studying the involvement of Pgps improved dsRNA triggers. Larval worms in parasitic biological process that can lead were exposed to dsRNA during in vitro to develop new way to control nematode culture or by injecting infected mosquitos, parasites. and worms were tested for thermosensory defects using a linear thermal gradient. In addition to RNAi, we also treated larval worms with chemicals that are known to interact with predicted thermosensory effectors and tested for thermosensory defects. We

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 198 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS01.20 The Giant African Snail Achatina Alto), which requires confirmation, since in Fulica as Carrier of Nematodes of Medical bordering municipalities this species is found and Veterinary Importance in the Rio De in dense populations, including specimens Janeiro State, Brazil infected with nematodes. This survey reinforces the importance of A. fulica in the Rodrigues1, Mônica Fernandez1, Suzete transmission of parasitic diseases of medical Rodrigues1, Langia Montresor2, Prof. Norma and veterinary importance, besides providing Labarthe3, Silvana Thiengo1 subsidies to the control and prevention of 1Laboratório de Referencia Nacional para those parasites. Esquistossomose-Malacologia - Instituto Oswaldo Cruz/Fiocruz, Brazil, 2Moluscário Lobato Paraense, Instituto de Pesquisas René PS01.21 Expression of Four Nicotinic Rachou/FIOCRUZ, Brazil, 3Fundação Oswaldo Acetylcholine Receptor Subunits in Ascaris Cruz, Brazil Suum Intestine and Muscle Cells

Invasive land molluscs have generated Mark McHugh1, Paul Williams1, Saurabh Verma1, economic damage to agriculture, environment Melanie Abongwa1, Shivani Choudhary1, and problems to animal and human health Sudhanva Kashyap1, JoAnne Powell-Coffman1, worldwide, including Brazil. Achatina fulica Alan Robertson1, Richard Martin1 is an intermediate host of nematodes of 1Iowa State University, United States medical and veterinary importance, such as Angiostrongylus cantonensis, A. costaricensis Ascaris suum is a gastrointestinal parasite and Aelurostrongylus abstrusus. Achatina of pigs which is of primary health concern fulica was introduced for commercial and economic importance to the swine purposes in Paraná State, South Brazil, in the industry. In the absence of effective vaccines, 1980’s and is currently recorded in all Brazilian anthelmintics are required for prophylaxis states, except in Rio Grande do Sul. Our main and treatment. However, intensive use has objective was to evaluate the occurrence of A. led to the development of resistance globally. fulica infected with Metastrongyloidea larvae Drugs such as levamisole and pyrantel in the 16 municipalities of the Mesoregion target nicotinic acetylcholine receptors Centro Fluminense of Rio de Janeiro state, (nAChRs) found on the somatic muscle of Southeast Brazil. Molluscs were collected the parasite. nAChRs are transmembrane from December 2016 to October 2018. In all, proteins comprised of five subunits that 358 A. fulica specimens were collected and surround a central cation-permeable pore examined through the artificial digestion and facilitate coordinated muscle movement technique. Morphotypes of nematode that is crucial for the parasite’s maintenance larvae were obtained: Angiostrongylus sp., of position in the intestine. Additionally, Rhabditis sp., Caenorhabditis sp. and A. different combinations or stoichiometry of abstrusus. Metastrongyloidea larvae are being nAChR subunits gives rise to a wide diversity identified using DNA Barcode (mitochondrial of receptor pharmacology. In vertebrates, cytochrome c oxidase subunit I). The nAChR function is not restricted to fast- municipality of Paraíba do Sul presented the neurotransmission. Given the diversity highest infection rate (72%), and Macuco of nAChR subtypes in nematodes, we (first report of A. fulica) presented the hypothesize that nAChR expression (and lowest infection rate (8%). Aelurostrongylus function) will not be restricted to nervous abstrusus larvae, a feline lung parasite, and muscle tissue in worms. Here we focus was observed in 44% of the investigated on the nematode intestine, which is involved municipalities. We highlight that the high in digestion and absorption of nutrients prevalence of this species indicates that the making it an attractive site for anthelmintic parasite may be underdiagnosed in cats, intervention. PCR was used to confirm and that A. fulica may be the main vector expression of nAChR subunits: Asu-unc-38, for its transmission. In four municipalities, Asu-unc-29, Asu-unc-63 and Asu-acr-8 that no specimens of A. fulica were found (Areal, constitute the putative levamisole receptor Bom Jardim, Duas Barras e São Sebastião do in adult female A. suum muscle and intestine.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 199 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Expression of these subunits within the calculation of the LC50 with the independent intestine was notable and unexpected. variables (dose) transformed by natural We conducted further experiments using logarithm (log dose). LC50 of citronellol was RNAscope in situ hybridization to localize 1.08 mg/mL demonstrating better efficacy of expression of the subunits at higher resolution this compound in inhibiting the development in the intestine and muscle; and qPCR of larvae in comparison to citronellal which to compare mRNA levels in both tissues. LC50 was 3.57 mg/mL. We concluded Calcium imaging was also conducted on citronellol showed high anthelmintic efficacy isolated intestinal tissue where levamisole against resistant strains of Haemonchus and acetylcholine elicited intracellular calcium contortus. - FAPESP/2018-02423-0 responses. These findings raise questions on the functional roles of nAChRs in the intestine which may not be limited to neuromuscular PS01.23 In Vitro Anthelmintic Effect of Four transmission, but an acetylcholine paracrine Extracts Obtained From Caesalpinia Coriaria function. Further studies on the mechanisms Foliage Against an Haemonchus Contortus involved in intestinal nAChR signaling is Isolate. 1. Egg Hatch Test paramount for therapeutic exploitation. Dr. Diana Rojas1, Dr Andrea Montenegro1, Dr. Leyla Rios de Alvarez1 PS01.22 Anthelmintic Effectiveness of 1AGROSAVIA (Corporación Colombiana de Citronellal and Citronellol on Haemonchus Investigaciones Agropecuarias), Colombia Contortus Evaluated In Vitro Larval Development Test The failure of anthelmintics to control parasites of small ruminants is a worldwide Poliana Pacheco1, Jessica Ribeiro2, Bruna problem, alternative control and plants Wedy2, Danielle Balbino2, Guillherme Luciani2, secondary metabolites are one of the Dr. Cecília Veríssimo2, Dr. Rodrigo Giglioti2, Dr. potential solutions. The present study Helder Louvandini1, Dr. Luciana Katiki2 evaluated the in vitro anthelmintic effect of 1University of São Paulo, Brazil, 2Institute of extracts obtained from Caesalpinia coriaria Animal Science, Brazil foliage, a legume consumed by sheep and goats in the department of La Guajira- Research on natural products is expanding Colombia, and used for medicinal purposes due to the promising results found in the by the Wayúu community. To measure the sustainable and strategic control against susceptibility of an Haemonchus contortus gastrointestinal nematodes. Citronellal and isolate the in vitro Egg hatch test was used Citronellol are natural acyclic monoterpenes with extracts of different solvents. The yields found in essential oils. The objective of of the extracts were 30.54, 23.51, 27.7 and this study was to evaluate the activity of 16.6% for acetone-water (ACW), methanol- these compounds on larvae of Haemonchus water (MW), acetone-dichloromethane-water contortus resistant to multiple drugs through (ADCW) and methanol-dichloromethane- the larval development test and determination water (MDCW), respectively. The anthelmintic of lethal concentrations (LC) using doses activity of the different concentrations ranging from 5 mg/mL to 0.00001 mg/mL. (125, 250, 500, 1000, 2000 and 3000 g/ For each concentration, 6 replicates were ml) of C. coriaria, were compared with the performed diluted in Tween and negative hatching percentage in Phosphate Bufferedμ control with Tween and distilled water. The Saline (PBS) as a negative control. Death eggs were placed in 48-well plates and of H. contortus was determined within a incubated for 24 hours at 27 °C. On the next period of 48 h. C. coriaria extract had mean day, nutrient medium and oils concentrations mortality of 46.7, 37.1, 38.8 and 29.8 % at were added and they were incubated more 5 4000 g/ml for the ACW, MW, ADCW, and days, until the larvae reached the third stage. MDCW, respectively; the extracts inhibited The counting of L3 and undeveloped larvae egg hatchabilityμ of H. contortus in a was evaluated under the inverted microscope. concentration-dependent manner. Effective The SAS Probit Program was performed for concentrations 50% (EC50) were calculate by

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 200 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Probit Analysis and best-fit values were 6763, generalized linear model with the log link. 8853, 8762 and 12872 μg/ml, for ACW, MW, Extract by exposure time (ET) interaction ADCW and MDCW, respectively. The ACW effect was statistically significant (P<0.05). was the most active against H. contortus At an ET of 10 minutes, there were not eggs, with the lowest EC50. This study statistically significant differences between demonstrates that the extracts of C. coriaria extracts, while at 60 minutes, all pairwise foliage have anthelmintic activity; therefore, differences were significant except for MW they could have an application in the control and MDCW, which yielded the lowest means. of helminths in livestock. Thus, larvae exsheathment was affected by extracts, but the extend of the effect depends on exposure time. At an ET of 60 minutes, PS01.24 In Vitro Anthelmintic Effect of Four EC50 values were 5927.4, 9955.4, 2883.4, Extracts Obtained From Caesalpinia Coriaria 9876.3 g/ml, for ACW, MW, ADCW and MDCW, Foliage Against An Haemonchus Contortus respectively. These results suggest that at this Isolate. 2. Larvae Exsheathment Inhibition Test ET, the μmost effective extract was ADCW.

Dr. Diana Rojas1, Dr. Jaime Cubides1, Andrea Montenegro1, Carlos Martínez2, Leyla Rios1 PS01.25 Anthelmintic Activity of a Synthetic 1Agrosavia (Corporación Colombiana Peptide Bioinspired in the Protein Rc-2S-Alb De Investigaciones Agropecuarias) CI Towards Haemonchus Contortus Tibaitatá, Colombia, 2Agrosavia, Dirección de Investigación y Desarrollo, Sede Central., Alexandra Martins Santos Soares1, Lucas Colombia Pinheiro Dias2, Pedro Filho Noronha Souza2, Ruana Cardoso Lima3, Luciana Magalhães The increase of anthelmintic resistance Rebelo Alencar3, Carolina Rocha Silva4, José worldwide, has caused researchers to focus Tadeu Abreu Oliveira2, Livio Martins Costa- on the use of plants in order to reduce the Junior4 use of commercial drugs. The present study 1Federal University of Maranhão, Brazil, evaluated the in vitro anthelmintic effect of 2Federal University of Ceará, Brazil, 3Federal extracts obtained from Caesalpinia coriaria, University Of Maranhão, Brazil, 4Federal a legume consumed by small ruminants in University Of Maranhão, Brazil the department of La Guajira-Colombia, used by the Wayúu indigenous community for The nematode Haemonchus contortus has controlling parasites of sheep and goats. The been developed resistance to the most effect was measured against an Haemonchus common drugs commercially available. In contortus isolate, the larvae exsheathment the search for alternatives to conventional inhibition test was performed, using L3 larvae anthelmintics, peptides have been (1 to 3 weeks) incubated for 3 hours in four investigated. Here, a linear synthetic peptide extracts of C. coriaria; then washed with named RcAlb-Pep bioinspired from the Phosphate Buffered Saline (PBS) solution antimicrobial protein Rc-2S-Alb was designed, and placed in 24-well plates to be exposed synthesized, and tested against H. contortus. to a hypochlorite solution (NaClO) at 0.05 Physicochemical properties of the peptide, % (previously standardized). Six repetitions the 3D structure model, the egg hatch per concentration (125, 250, 500, 1000, inhibition and larval development inhibition 2000 and 3000 g/ml) were used and a of H contortus were carried out. Additionally, negative control in PBS. The extracts were the ultrastructure of the nematode after prepared with differentμ solvents: acetone- treatment with the peptide was evaluated by water (ACW), methanol-water (MW), atomic force microscopy. The peptide showed acetone-dichloromethane-water (ADCW) molecular weight, Boman Index, hydrophobic and methanol-dichloromethane-water ratio and net charge of 826.05 Da; -1.28; (MDCW). Using PBS control, 100% of larvae 62%; and +1; respectively. RcAlb-Pep has a were exsheathed at 60 min after adding small negatively charged region and a larger NaClO. Counts of non-exsheathed larvae positively charged region, due the presence were analyzed using a negative binomial of positively charged amino acid lysine

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 201 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

(Lys). The RcAlb-Pep inhibited the larval by AFM. H. contortus eggs, L1, L2, L3 and development of H. contortus with an EC50 of adults females were obtained from sheep 0.15 mg mL-1 and had no effect on egg hatch. that were experimentally infected. The L3 Atomic force microscopy reveals the affinity larvae were unsheathed with 0.02 % sodium of RcAlb-Pep with the cuticle of H. contortus hypochlorite solution. For fixation, all stages in L2 stage. For the L3 stage, low affinity with of H. contortus were maintained in 5% RcAlb-Pep was verified, which indicates the formalin. We observed a qualitative reduction low interaction is due to the formation of in Young’s modulus when eggs develop the sheath at this stage, promoting an extra from the morulae to the larvae stage. The protection against biomolecules targeting AFM analysis of L1 stage showed a series of the cuticle. In conclusion, the bioinspired periodically separated annuli, with remains RcAlb-Pep has potential to be used as a from larvae hatching on the nematode cuticle new anthelmintic compound to control of at this stage. The images of H. contortus gastrointestinal nematode parasites. in adult, L3 with sheath, L2 and L1 stages make it possible to compare the alterations of the annuli structures during the evolution PS01.26 Assessment of Biophysical of the parasite. The artificially unsheathed Properties of Haemonchus Contortus From process made evident the genital primordium Different Life Cycle Stages With Atomic of H. contortus in the L3 stage. The results Force Microscopy revealed an increase in adhesion values on the surface of the nematode in the L3 stage due Ms. Carolina R. Silva1, Dr. Livio M. Costa- to sheath removal. Our results provide early Junior1, Dr. Alexandra M. S. Soares2, Dr. Alan S. insight into the differential biomechanical Menezes3, Dr. Maria R. L. Silva4, Dr. Alessandro and ultrastructural properties of our samples, F. T. Amarante4, Dr. Luciana M. R. Alencar3 which can explain biological and biochemical 1Laboratory of Parasite Control, Department steps in the life cycle of these parasites. of Pathology, Center for Biological and Health Sciences, Federal University of Maranhão, Brazil, 2Chemical Engineering Course, Center PS01.27 Anthelminthic Effect of Extracts for Exact Sciences and Technology, Federal from Petiveria Alliacea and Diospyros University of Maranhão, Brazil, 3Department Anisandra on Haemonchus Placei Larval of Physics, Federal University of Maranhão, Development and Egg Hatching Brazil, 4Bioscience Institute, São Paulo State University, Brazil Gabriela Janett Flota Burgos1, Dr. José Alberto Rosado Aguilar1, Dr. Roger Iván Rodríguez Haemonchus contortus is the most Vivas1, Dra. Rocío de Lourdes Borges Argáez2, pathogenic and economically important Dra. María Marcela Gamboa Angulo2, Dra. gastrointestinal nematode species infecting Cintli Martínez Ortiz de Montellano3 small ruminants. Light, scanning and 1Universidad Autónoma De Yucatán, Mexico, transmission electron microscopy have 2Centro de Investigación Científica de been successfully used to assess damage Yucatán, México, 3Universidad Autónoma de or structural parameters of H. contortus. México, México However, some limitations of these techniques are preventing advances in understanding Haemonchus placei is one of the most the effect of different compounds on H. prevalent and pathogenic nematodes that contortus. Atomic Force Microscopy (AFM) is affect cattle. Several anthelminthics have been a high-resolution microscopy technique that used for its control; however, resistance to provides information about the topography benzimidazoles, levamisole and macrocyclic and surface composition of a wide variety of lactones has been reported. Control materials ranging from living individual cells alternatives include the use of plant extracts through fixed tissue. The present study report, with anthelminthic effect. The objective of for the first time, assessed the topographic the present study was to evaluate the in vitro and biomechanical characterization of H. anthelminthic effect of Petiveria alliacea and contortus eggs, larvae and adult cuticle Diospyros anisandra extracts on Haemonchus

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 202 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

placei larval development and egg hatching. which might impair the effectiveness of a Extracts from P. alliacea (stems and leaves) treatment. The objective of this study was and D. anisandra (bark and leaves) collected to assess the in vitro AH-like activity of PCs- in dry and rainy season, were evaluated by combinations (Coumarin:Quercetin, and the egg hatch assay (concentrations: 600 g, Cafeic-acid:rutin; 8:2 ratio) against adult 300 g, 150 g, 75 g and 37.5 g per ml). Lethal motility of the cattle nematode C. punctata. concentrations at 50% (LC50) were obtainedμ Immediately after recovery, 10-13 motile C. withμ Polo plusμ software.μ The μextracts from punctata adult worms were incubated at stems and bark had higher percent of egg 37 °C with the PCs-combinations for 2, 4, 6, hatch inhibition (EHI) than the extracts from 12 and 24 hours, with a 5 % CO2 inclusion. leaves. Extracts from P. alliacea stem and Five increasing concentrations were used for D. anisandra bark, collected in rainy season Coumarin:Quercetin (0.043, 0.087, 0.175, 0.35, showed higher percent of EHI (91.9% and 0.7 and 1.4 mg / mL -1) and Caffeic-acid:Rutin 96.4% in 300 g/ml, respectively) than the (0.052, 0.105, 0.21, 0.42, 0.84 and 1.68 mg / extracts collected in dry season. The extracts mL -1). Ethanol-2.5 % and physiological saline of P. alliacea stemμ and D. anisandra bark solution were used as negative controls. Four collected in rainy season showed the lowest replicates were run for each concentration LC50 (78.9 and 71.6 g/ml, respectively). and control. Individual motility was calculated Additionally, it was demonstrated that these as: Worm motility%= 100 (motile worms in extracts have high ovicidalμ effect (91.6% and the concentration / total number of worms 89.0% in 300 g/ml, respectively). The results in the same concentration), corrected for by of the present study showed that the extracts control motility percentages. Motility of worms of stem from P.μ alliacea and bark from D. was not affected after 2 and 4 hours using anisandra collected in rainy season have high both PCs-combinations, neither for 6 hours anthelmintic effect in vitro on Haemonchus post-exposure to Caffeic-acid:Rutin (P>0.05). placei larval development and egg hatching, Estimated EC50 for Coumarin:Quercetin were: representing a possible alternative for the 1.621 ± 0.154 mg mL-1, 0.398 ± 0.064 mg mL-1 control of these nematodes. and 0.073 ± 0.071 mg mL-1 for time lapse of 6, 12 and 24 hours, respectively. Caffeic acid:Rutin EC50 estimation was: 0.192 ± 0.061 mg mL-1 PS01.28 Anthelmintic-like Activity of Four and 0.051 ± 0.164 mg mL-1 for time lapse of Polyphenolic Combinations and Their 12 and 24 hours, respectively. Results suggest Interactions Against Adult Worms Cooperia that PCs-combinations assessed interfere Punctata nematodes neurophysiology which leads to their paralysis. After addressing their potential Dr. Elke von Son-de Fernex1, Estefanía toxicity and pharmacokinetics, formulations Zúñiga-Olivos1, Dr. Miguel Ángel Alonso-Díaz1, could be considered for in vivo field trials. Dr. Pedro Mendoza-de Gives2, Dr. Epigmenio Castillo-Gallegos1 1Centro de Enseñanza Investigación y PS01.29 Prevalence of Gatrointestinal Extensión en Ganadería Tropical, Facultad de Parasites and the Effective Control Trials in Medicina Veterinaria y Zootecnia, Universidad Some Dairy Farms in Bahrain Nacional Autónoma de México, Mexico, 2Centro Nacional de Investigaciones en Dr. Abdalla Fadlalla Azrug Ahmed, Eman Parasitología Veterinaria, INIFAP, México Magzoub Saeed1, Abdelaziz Atta M. Alatta1 1National Veterinary Laboratory, Animal Health Polyphenolic compounds (PCs) from bioactive and Control Directorate, Agriculture and plant extracts have been identified and Marine Resources Affairs, Bahrain assessed for anthelmintic (Ah)-like activity. Previous reports have proposed the use of Livestock represents an important PCs-combinations (Coumarin:Quercetin and contributive part of the economy, although Caffeic-acid:Rutin) against free-living stages the livestock population is very small due to of Cooperia punctata. However free-living and the limited land-space in Bahrain. This field parasitic stages have biological differences, study was conducted to evaluate the parasite

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 203 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

status of dairy cattle in Bahrain. Material and was based on the mean score per group at Methods: A clinical epidemiological study in each time point and assessed by the Abbott’s dairy cattle was conducted in Bahrain during formula: 93.6% on D2 and 100% from D8 until 2017 - 2018. The study evaluated fresh fecal D28 (p<0.0001). samples collected from a total of 92 cattle across 11 dairy farms. The study included The second study was conducted in a various animal ages, breeds, lactation stages commercial laying farm in Brazil. The Isa and sexes to study the significance on Brown laying hens were housed in 2 similar parasitic infestivity across animal breeds and buildings of ca.5000 hens and were infested sexes. Results: The overall prevalence rate with the Poultry Red Mite (Dermanyssus of Haemonchus contortus among sampled gallinae). One house was treated and the cattle was 26.07% (24 of 92 samples tested). other left untreated. The infestation was Most of the positive cases were found as evaluated in blind conditions using mite traps mixed infections with other species of (20 per house left 48h on 12 occasions over gastrointestinal parasites, Trichostrongylus 4 months). Treatment efficacy was based species , Trichuris, Nematodirus and on the mite reduction in traps (Henderson- Eimeria species while all these species were Tilton formula). It reached 95.9% on D2 and considered as less significant due to the level 100% from D8 till D115. A positive effect was of infection. Eimeriosis caused by various observed on hen mortality and laying rate. Eimeria species was seen in 17 cases (18.5%) The third study was conducted in a causing serious losses among newly born commercial broiler breeder farm in the calves. Discussions and Conclusion: The Philippines of ca.11,000 chickens housed in effects of gastrointestinal helminthes among cages. The visual examination of the vent cattle in Bahrain were varied with animal and the feathers of 79 randomly selected ages, management and nutritional conditions, birds revealed a multiple infestation with lactating stages and severity of infestations. Tropical Fowl Mites (Ornithonyssus bursa) and feather mites (Megninia cubitalis, Pterolichus obtusus). No mites were seen on these birds PS01.30 Clinical Efficacy of Exzolt® (1% from D7 till D56. Fluralaner Aqueous Solution) In the Treatment of Mite Infestations of Flocks In These 3 studies showed the fast onset and Tropical Countries high efficacy of Exzolt® against not only 3 species of blood-sucking poultry mites but Emmanuel Thomas1, Rafael Chiummo1, Luis also against feather mites. Vettorato2, Luis Alves3, Nathaniel Mendoza Realeza4, Anja Heckeroth1 1MSD Animal Health, Germany, 2MSD Animal PS01.31 Evaluation of DynaTrap Flylight (DT- Health, Brazil, 3State University of Western 3009) Plug-In Units to Trap Indoor House Paraná (UNIOESTE, Brazil, 4MSD Animal Flies and Stable Flies Health (Phils) Inc, Philippines Dr. Jerome Hogsette1, Dr. James Cilek2 Three clinical studies were conducted in Brazil 1USDA/ARS, United States, 2Navy Entomology and the Philippines in flocks affected with Center of Excellence, United States different fowl mites. All flocks were treated with Exzolt® at the label dose of 0.5 mg/kg DynaTrap Flylight traps with 9-watt UV BW in drinking water twice 7 days apart, D0 lights were evaluated alone and paired with and D7. Gardner GT-200 traps, an industry standard. Gardner traps are larger than DynaTraps with The first study involved 60 Hisex Brown two 40-watt fluorescent tubes. Evaluations laying hens sourced from a commercial were conducted in a windowless room at farm in Brazil. All had a Northern Fowl Mite 23°C with constant illumination. Tests used infestation, Ornithonyssus sylviarum (score colony-reared house or stable flies. Flies were ≥ 2 of 7). They were separated into 2 groups, anaesthetized and used after a 30-minute treated and untreated. The treatment efficacy recovery period. Trap pairs were placed

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 204 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

parallel to each other, 2 m apart and 1 m high PS01.33 First Detection and Molecular on a test room counter. Single traps were Identification of Neospora Caninum From placed on the same counter with other traps Naturally Infected Cattle, Sheep and Goats in removed. Captured flies were recorded at 1, North Africa 4, and 24 hours post-release. After the 24- hour count, remaining live flies were killed and Yosra Amdouni1, Mohamed Ridha Rjeibi1, Safa glue boards were replaced. Single and paired Amairia1, Yousra Said1, Sofia Awadi2, Prof. traps were tested over 6 dates and traps in Mourad Rekik3, Prof. Mohamed Gharbi1 paired tests were rotated after every replicate. 1Laboratoire de Parasitologie, Univ. Because the DynaTrap is designed to plug Manouba, Institution de la Recherche et de into outlets in walls, tests were performed l’Enseignement Supérieur Agricoles, École with a simulated wall. At 24 hours, individual Nationale de Médecine Vétérinaire de Sidi DynaTraps captured about 83% and 33% of Thabet, Tunisia, 2Regional Slaughterhouse the house flies and stable flies, respectively. of Béja, Tunisia, 3International Center for These results changed slightly with the Agricultural Research in the Dry Areas simulated wall placed behind the units. In (ICARDA), Jordan both cases, significantly more house flies were trapped than stable flies. In paired tests, Neospora caninum is a coccidian parasite Gardner traps captured significantly more with a wide range of hosts. It is considered house flies (93%) and stable flies (55%) at 24 as a major cause of reproductive failure in h than the DynaTraps (house flies = 4%, stable livestock females causing economic losses. flies = 9%). Gardner traps, tested individually, The objective of this study was to estimate captured significantly more house flies than the infection rate and molecular identification stable flies. Single DynaTraps effectively of N. caninum in Tunisian ruminants. captured house flies indoors, but were less A total number of 469 meat samples effective against stable flies, which are not were collected from 150 cows, 198 ewes usually found indoors. The Gardner GT-200 and 121 goats slaughtered in the regional captured more flies than DynaTraps because slaughterhouse of Béja and tested for the of its size and brightness. The comparison presence of N. caninum ITS1 gene using between these traps provides an indication of PCR followed by sequencing of some PCR differential capture rates based on brightness. products. The three nucleotide sequences The DynaTrap performed well and removed are available in the GenBank database under house flies from the environment at a better- the accession number KY496699 for cattle, than-expected rate. KY562727 for sheep and KY486651 for goats. A phylogenetic tree was then constructed. The overall molecular infection prevalence of PS01.32 Flight Time and Distance N. caninum was significantly higher in cattle Comparison of Field Collected and Colony than in goats and sheep (22, 19 and 10.6%, Reared Horn Fly and Face Fly respectively, p=0.001). In sheep, the highest prevalence was observed in the northern Béja Wes Watson1, Fallon Fowler, Steve Denning (31.2±16.1), with the Noire de Thibar breed as 1Department of Entomology and Plant the most infected sheep breed (31.7±14.2%) Pathology, NCSU, United States (p<0.001). In cattle, and goats, there were no differences in the molecular prevalence of N. Flies reared in captivity may exhibit a fitness caninum according to breeds and localities. cost, reducing flight time and distance when The association between age and N. caninum compared to wild type flies. To measure molecular prevalence was statistically potential fitness cost, we compared the significant in the three species; the highest flight time and distance of horn fly and face prevalence was observed in cattle between fly using flight mill instrumentation. First two and eight years of age (28.8±10.9%), generation flies emerging from field collected in goats aged between two and four years dung pats and from laboratory reared (31.9±13.3%) and in sheep of more than one colonies under teneral, 2d, and 5 day old year of age (19.4±9.1%). Comparison of the conditions were compared. partial sequences of the ITS1 gene revealed

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 205 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

96-100% similarity with amplicon deposited in The cyst counts were reduced by > 90% from GenBank. day 3 to 5 (86.8%, 83.7%, 99.97%, 99.97%, 93.5%, from day 1 to day 5 respectively) and To our knowledge this is the first detection the trophozoite counts by 99.9% in the dogs and molecular identification of N. caninum in treated compared to placebo group (192106 in cattle, sheep and goats in North Africa. This placebo vs 183 in treated group). information is pertinent in designing control programmes that would reduce economic These results indicate that the metronidazole losses in the livestock industry. suspension was effective for the treatment of Giardia infection (cysts and trophozoites) in dogs and demonstrate the link between the PS01.34 A Pilot Study to Determine the reduction of Giardia cysts in faeces and of the Efficacy of a Metronidazole Suspension trophozoite forms in the gut in metronidazole on the Reduction of Cyst and Trophozoite treated dogs. Counts in Dogs Infested with Giardia

Dwight Bowman1, Céline Nicolas2, Christelle PS01.35 Approaching to Development of Navarro2, Sandrine Fournel3, Sloane Joanes4, Transmission Blocking Vaccine Based on Dr. Vanessa Chala2 Developmental Stages of Babesia Gibsoni 1Cornell University, United States, 2Virbac Global Medical, France, 3Virbac R&D, France, Dr. Seung-Hun Lee1, Prof. Xuenan Xuan1 4Virbac R&D, United States 1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and The objective of this randomized masked Veterinary Medicine, Japan study was to determine the effectiveness of an oral metronidazole suspension (Eradia/ Babesia gibsoni is a tick-borne Ayradia, Virbac), when used in dogs infested hemoprotozoan pathogen and it may cause with Giardia spp., to reduce Giardia cysts and canine babesiosis. Clinical signs of canine trophozoites. babesiosis include fever, thrombocytopenia, anemia, and sometimes can be fatal, however, From the eighteen dogs screened (> 3 month- effective vaccine or treatment are not old, from 5-10 kg), ten were positive for available yet. Recently, transmission blocking Giardia and six dogs with the highest fecal vaccine that aims to prevent transmission cyst counts (≥ 750 cysts per gram, determined of pathogens from infected to un-infected by immunofluorescent assay) were selected. host has been studied. The purpose of this The dogs were evenly and randomly allocated study is to develop transmission blocking to either a placebo (control) group or a vaccine based on different gene expression metronidazole treated group (25 mg/kg of level of B. gibsoni in blood stage and in tick body weight BID) for five consecutive days. stage. First, from B. gibsoni (Oita strain) Fecal samples were collected once daily and blood stage cultured in vitro, ring form and submitted for cyst counts. At the end of the free merozoites were separated using Percoll study, samples of the entire small intestines density gradient centrifuge. The separated were sent to a laboratory for the trophozoite parasites were in alive after centrifuge. count. The number of trophozoites recorded However, syringe filtering, refrigeration, and for each intestinal section for each dog were sorbitol treatment that are known can be summed and the resulting number was the used for developmental stage separation of total number of trophozoites per dog. Efficacy Plasmodium spp. did not applicable on B. of treatment was measured as a reduction gibsoni. On the other hand, to separate B. in the number of both Giardia spp. cysts gibsoni in tick stage, semi-artificial feeding (Henderson-Tilton formula) and trophozoites system which uses mouse skin instead of dog after treatment (geometric means). was used. As a vector tick, Haemaphysalis No serious adverse events were reported longicornis (Okayama strain) which is known during the study. vector of B. gibsoni was used. Using semi- artificial feeding system, H. longicornis was

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 206 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

infected with B. gibsoni in midgut and ovary PS01.37 Optimized Excystation Protocol for which was confirmed by PCR. Morphology Ruminant Eimeria Spp. Sporulated Oocysts and transcriptome of each developmental (Apicomplexa, Coccidia) stages will be analyzed. Sara Lopez Osorio1, Dr. Liliana M.R. Silva3, Dr. Zahady Velazquez33, Dr. Habil Anja Taubert3, PS01.36 Establishment of a Stable Dr. Habil Carlos Hermosilla3 Transfection System for Genetic Manipulation 1Universidad De Antioquia - Justus Liebig of Babesia Gibsoni University, Colombia, 2CIBAV Research Group, Facultad de Ciencias Agrarias, Dr. Mingming Liu1, Dr. Xuenan Xuan1 Universidad de Antioquia, Colombia, 3Institute 1Obihiro University of Agriculture and for Parasitology, Justus Liebig University, Veterinary Medicine, Japan Germany

Aims: Genetic manipulation techniques, such An improved method for excystation and as transfection, have been previously reported collection of infective sporozoites from in many protozoan parasites. In Babesia, stable sporulated oocysts of E. bovis and E. arloingi, transfection systems have only been established is here described. The protocol uses conditions for bovine Babesia parasites. We recently which mimics the intestinal anaerobic reported a transient transfection system and environment of ruminants in vivo. Firstly, the selection of promoter candidates for sporulated oocysts were washed with 6% Babesia gibsoni. The establishment of a stable sodium hypochlorite solution. Then, Eimeria transfection system for B. gibsoni is considered oocysts have been filtered through different to be urgent to improve our understanding of sieves sizes to remove debris with a recovery the basic biology of canine Babesia parasites percentage of 95%. Afterwards, the oocysts for a better control of babesiosis. have been treated for at least 12 h with 0.02M L-cysteine HCl/0.2M NaHCO3 solution in a Contents: To establish a stable transfection 100% CO2 environment at 37 °C. The last system, we generated a plasmid construct in oocyst treatment was performed with a 0.4% which the 5 -intergenic (IG) region-B of the ef- trypsin/8% sterile bovine bile excystation 1 gene (5 -ef-1 ) drives the gfp reporter gene, solution, which disrupted oocyst wall thereby and the 5 -actin′ promotes the expression releasing up to 90% of sporozoites in ofα the selection′ α marker hdhfr. The plasmid approximately 2 h of incubation (37 °C ) with was designed′ for integration into the ef-1 a 1:2 (oocysts:sporozoites) ratio. Free-released locus of B. gibsoni genome by double cross- sporozoites were up to 99% viable, highly over homologous recombination. Linearizedα motile, capable of infect host cells and to form plasmid was transfected by 4D Nucleofector™ meront stages in vitro. The improved method into in vitro cultured B. gibsoni and 10 nM is much cheaper, faster, and accessible for all WR99210 was added for drug selection labs with minimum equipment, and without two days after transfection. GFP-expressing requirement of neither expensive solutions nor parasites were observed by fluorescence sophisticated instruments. microscopy as early as two weeks after drug selection, and consistently expressed GFP for more than 3 months without drug pressure. Genome integration was confirmed by PCR, sequencing and Southern blot analysis.

Conclusions: We present the first successful establishment of a stable transfection system for B. gibsoni. This finding will facilitate functional analysis of Babesia genomes using genetic manipulation and will serve as a foundation for the development of tick- Babesia and host-Babesia infection models.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 207 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS01.38 Molecular Characterization of PS01.40 Targeted Overexpression of Cyclic Cryptosporidium in Calves From Rural AMP-Dependent Protein Kinase Subunit in Settlements in the Northwest Region of the Toxoplasma Gondii Promotes Replication State of São Paulo, Brazil and Virulence in Host Cells

Lucas Vinicius Shigaki de Matos1, Luiz Silveira Dr. Hongchao Sun1, Dr. Yi Yang1, Prof. Aifang Neto2, Bruno César Miranda Oliveira5, Miriam Du1 Yumi Makatu5, Julia Cestari Pierucci1, Milena 1Institute of Preventive Veterinary Medicine, Araúz Viol5, Jancarlo Ferreira Gomes3, Gilson College of Animal Sciences, Zhejiang Pereira de Oliveira1, Giovanni Widmer4, University, China Marcelo Vasconcelos Meireles5, Alex Akira Nakamura5, Elis Domingos Ferrari5, Katia Toxoplasma gondii (T. gondii) is one of the Denise Saraiva Bresciani5 most common parasites that can infect 1Faculdade de Ciências Agrárias, FCAV/ almost any warm-blooded animals including UNESP, Brazil, 2Universidade Federal do humans. The cyclic nucleotide-dependent Tocantins, UFT, Brazil, 3Universidade Estadual protein kinase (PKA) regulates a spectrum de Campinas, Unicamp, Brazil, 4Cummings of intracellular signal pathways in many School of Veterinary Medicine at Tufts organisms. Protein kinase catalytic subunit University, United States, 5Faculdade de (PKAC) is the core of the whole protein, Medicina Veterinária de Araçatuba, FMVA/ and plays an important role in the life cycle UNESP, Brazil of T.gondii. Here, T.gondii PKAC (TgPKAC) overexpression strain (TgPKAC-OE) was The study was conducted on 25 properties constructed. The growth of the TgPKAC-OE, of the settlements São José I and Salvador, RH Ku80, and TgPKAC inhibition strains located in the municipalities of Brejo Alegre (TgPKAC-H89) were analysed by SYBR- and Birigui, in the State of São Paulo, Brazil. green△ real-time PCR, and the ultrastructure A record of variables was elaborated and was observed by transmission electron included data such as gender, breed and age microscopy. The survival rate in mice was of the animals. A total of 231 stool samples also recorded to analyse the virulence of the were collected from bovines aged one to six parasites. We also investigated the subcellular months, 128 being females and 103 males, localization of TgPKAC in Vero cells by laser 131 crossbred and 100 Holstein. Among the scanning microscope. We found that TgPKAC- 231 samples, 17 (7.36%) were positive for OE strain exhibited obviously increased Cryptosporidium spp. both by malachite growth rate in Vero cells in vitro, and infected green negative staining and by nested- mice survived for a shorter time compared to PCR. Of the 17 positive samples, 14 were wild type strain. Ultrastructural analysis found sequenced in agarose gel. These sequences more autophagosomes-like structures in were detected between 99% and 100% of TgPKAC-H89 parasite compared to RH Ku80 genetic similarity for the following species. strain, and the relative expression level of One sequence was similar to C. parvum Toxoplasma gondii autophagy-related protein△ (AB513880.1), one to C. bovis (MF074602.1), (ATG8) in TgPKAC-H89 parasite was higher two to C. ryanae (KT922233.1), one to C. felis than wild type parasite. Laser confocal results (KM977642.1) and nine were similar for C. showed that TgPKAC was mainly expressed andersoni reference MF350628. C. andersoni in the cytoplasm of Vero cells. In conclusion, was found in animals aged 2–6 months, an we hypothesized that inhibition of TgPKAC age group which is different from those could cause autophagy of Toxoplasma gondii described by several authors. The presence and then influence the replication of the of C. parvum indicates that the calves in parasite. TgPKAC plays an important role in the studied region should be considered a parasite virulence in vivo, and the subcellular potential source for zoonotic transmission. localization was successfully detected in Vero For the first time to our knowledge, C. felis cells. Our data will provide a basis for further was identified in cattle in America. study of TgPKAC function and help screen drug targets of T. gondii.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 208 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS01.41 Klossiella Equi: A Cause of Renal PS01.42 The Bacterial Microbiome of Disease in Horses? Rhipicephalus Sanguineus Ticks in the Mnisi Community, South Africa Elizabeth Zeldenrust1, Alexandre Leveille1, Dr. Karen Carleton1, Dr. Daniel Kenney1, Dr. John Miss Rebecca Ackermann1, Dr. Cory Gall2, Barta1 Prof. Kelly Brayton1,2, Prof. Nicola Collins1, Dr. 1Department of Pathobiology, Ontario Agatha Kolo1, Miss Jeanette Wentzel3, Dr Ilana Veterinary College, University of Guelph, van Wyk3 Canada 1Vectors and Vector-borne Diseases Research Programme, Department of Veterinary Klossiella equi (Apicomplexa, Adeleorina) is a Tropical Diseases, Faculty of Veterinary monoxenous coccidian parasite that infects the Science, University of Pretoria, South Africa, kidneys of equids globally. Usually, K. equi is 2Department of Veterinary Microbiology and an incidental finding during necropsy because Pathology, Washington State University, clinical signs referable to it are usually absent. America, 3Hans Hoheisen Wildlife Research However, gross and histological lesions have Station, Faculty of Veterinary Science, been reported from infected kidneys. It is University of Pretoria, South Africa unknown if K. equi contributes to or causes clinical kidney disease but long-term infection Rhipicephalus sanguineus is a three host tick has been documented in immunocompromised that completes all its life stages on domestic hosts. Although sporocysts that are presumed dogs. It is, however, capable of parasitizing to be immediately infective to other horses most vertebrates, including wildlife, cattle, are shed in the urine of infected animals, dogs and humans. It is known to transmit sporocysts are rarely detected by normal various tick-borne diseases. In Mnisi, a rural urinalysis; worse, sporocysts are difficult to community in Bushbuckridge, Mpumalanga, float even when targeted. Shedding periods South Africa, R. sanguineus is one of the most and duration of the lifecycle remain unknown. prevalent ticks found on dogs. The community With the difficulty of detecting the parasite lies at the wildlife-livestock-human interface antemortem, the true prevalence of K. where humans are at risk of infection with equi is unknown locally (Southern Ontario) various tick-borne zoonotic diseases. The aim and globally. With the recent publication of this study was to investigate the prevalence of sequences from K. equi (partial nuclear and diversity of tick-borne bacterial 18S rDNA; whole mitochondrial genome), pathogens in R. sanguineus that may impact detecting infections with K. equi in horses human and animal health. To achieve this, we antemortem may be possible using molecular analysed the microbiome of ticks sampled means. To test this, tissue, blood and urine from community dogs over a 12-month period samples are being obtained opportunistically to detect bacterial pathogens and symbionts. from horses submitted for routine necropsy to To date, R. sanguineus (n=582), R. simus the Animal Health Laboratory at the University (n=82), Amblyomma hebraeum (n=97), as well of Guelph, regardless of the underlying cause as 183 unidentified tick have been collected of death. For cases with fresh urine available, from 51 dogs. Ticks were kept in a humidity detection of sporocysts will be attempted and temperature controlled chamber for microscopically. DNA extracted from kidney two days to allow them to digest their blood tissues will be tested for the presence of K. meal. Three R. sanguineus ticks from each equi using PCR with species-specific primers dog were surface sterilized, and dissected to to establish the prevalence of this coccidium remove their midguts and salivary glands and in Southern Ontario. DNA from blood and then pooled. Genomic DNA was extracted urine from positive cases will be used to and PCR amplified using universal 16S rDNA explore the feasibility of developing a PCR barcoded primers. Sequencing will be done assay to detect K. equi antemortem in blood at Washington State University using Pacific or urine samples. Such a test would permit a Bioscience’s circular consensus sequencing better understanding of the prevalence of this strategy. Environmental conditions and parasite globally and its role, if any, in clinically other factors that could influence the tick relevant changes to the equine kidney. population or tick microbiome were also

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 209 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

analysed. Preliminary data shows that the PS01.44 Simulation Model of Host- microbiome is dominated by Coxiella spp, Transferring Adult Males Ticks (Dermacentor including C. burnettii, the causative agent Variabilis) (Acari: Ixodidae) in the of Q-fever.The information gained about Transmission of Equine Piroplasmosis the bacterial communities that abound in R. (Theileria Equi) sanguineus will undoubtedly aid health care practitioners in the area with the diagnosis of Mr. Taylor Donaldson1, Hsiao-Hsuan Wang2, important tick-borne diseases in animals and William Grant2, Pete Teel3 humans. 1Texas AgriLife, Texas A&M University - Dept. Entomology, United States, 2Texas A&M University - Dept. Wildlife and Fisheries PS01.43 Efficacy of a Single Spot-on Sciences, United States, 3Texas A&M Administration of Fipronil and Permethrin University - Dept. Entomology, United States Against Haemaphysalis longicornis in Dogs Equine piroplasmosis (Theileria equi) is a tick- Prof. Frederic Beugnet1, Wilfried Lebon1, borne disease of horses and other equines. Julian Liebenberg2 In 2009, an outbreak occurred in south 1Boehringer-ingelheim, France, 2Clinvet, South Texas, where Dermacentor variabilis were Africa collected off infected horses. Experimental transmission studies have shown D. variabilis The efficacy of fipronil plus permethrin to be a vector of T. equi by means of (Frontline Tri-Act®, Boehringer Ingelheim), intrastadial transmission, specifically by male administered once topically at the minimum adult ticks. Male ticks can remain on the hosts recommended dose, was assessed in dogs for long periods of time and take multiple experimentally infested with Haemaphysalis bloodmeals. Additionally, males may transfer longicornis ticks. from host-to-host which can allow for further transmission. Male transfer has been well The study was a blinded, negative controlled documented in D. albipictus with Anaplasma clinical efficacy study using a randomized marginale but no studies have investigated block design. Fourteen purpose bred Mongrel male transfer with D. variabilis. The aim of and/or Beagle dogs, 6 females and 8 males, this study was to model the transmission of were included. Dogs were randomly allocated T. equi while incorporating the transfer of either to the negative control group, or to males from one host to another. Simulations the treated group. Dogs were experimentally were run over a five-year period with four infested with approximately 50 viable, adult, different infection probabilities (1%, 0.5%, unfed female H. longicornis ticks (Okayama 0.25%, and 0.1%) and five transfer probabilities strain, Japanese origin) on days -2, 7, 14, of males (100%, 75%, 50%, 25%, and 1%). The 21 and 28. On Day 0, dogs in Group 1 were number of horses infected varied with the left untreated and dogs in Group 2 were combination of the two rates. The duration treated with Frontline Tri-Act® (fipronil plus of the infection was influenced by both rates. permethrin). Ticks were thumb counted at Results from this study show that transferring 24 hours after post-treatment infestations on adult male D. variabilis may play an important Days 8, 14, 22 and 29, and were removed and role in the transmission of T. equi. In addition, counted at 48 hours after treatment or tick this model may serve as a starting point for infestations on Days 2, 9, 16, 23 and 30. further investigation of the role of transferring Frontline Tri-Act® administered once male ticks in other disease systems. topically at the minimum recommended dose was highly effective against existing tick infestations (curative efficacy of 98.7%, p=0.0006). Regarding the re-infestations, the sustained efficacy of Frontline Tri-Act® was 100%, 100% 98.8% and 97.1% on Days 9, 16, 23 and 30, respectively. In addition, 24h in-situ count ranked from 87.2% to 100%.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 210 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS01.45 An Insight into Genetic Diversity infesting bovines across different AE zones of Ticks of Bovines Across Different Agro- of Pakistan. Findings of this study highlights Ecological Zones of Pakistan the importance of using morphological and molecular approaches for identification of ticks. Mr. Abdul Ghafar1, Prof. Robin B. Gasser2, Mr. Adil Khan1, Prof. Abdul Jabbar1 1Department of Veterinary Biosciences, PS01.46 Evaluation of Four Drug Application Melbourne Veterinary School, University Strategies for Rhipicephalus Microplus of Melbourne, Australia, 2Department of Control in Crossbred Naturally Parasitized Veterinary Biosciences, Melbourne Veterinary Cattle in Água Clara Municipality, MS, Brazil School, University of Melbourne, Australia Paulino Bonatte Junior2, Willian Maciel1, Introduction: Tick infestation is one of the Leandro Higa3, Mr. João Dillio6, Marcos Garcia4, leading causes of economic losses in bovines Jacqueline Barros5, Renato Andreotti5 across tropical and sub-tropical regions of 1Universidade Estadual Paulista - UNESP, the world. In Pakistan, bovine population is Brazil, 2Pós-Graduação em Ciência Animal, exposed to tick infestation throughout the year Faculdade de Medicina Veterinária, UFMS and little is known about biology and genetic - Univ. Federal Mato Grosso do Sul, Brazil, diversity of tick population across different 3Programa de Pós-Graduação em Doenças agro-ecological (AE) zones of the country. Infecciosas e Parasitárias, Universidade Federal de Mato Grosso do Sul, Brazil, Objective: To determine the genetic diversity 4Bolsista Fundapam/ Laboratório de Biologia of hard ticks infesting bovine population do Carrapato, Embrapa Gado de Corte, Brazil, across different AE zones of Pakistan. 5Empresa Brasileira de Pesquisa Agropecuária, Materials & methods: Thirty villages were Embrapa Gado de Corte, Brazil, 6Universidade selected from five different AE zones of do Oeste Paulista - Unoeste, Brazil Punjab and Sindh provinces of Pakistan and the sampling was carried out from September The study objective was to evaluate four to November 2017. A total of 753 adult ticks medication application strategies for tick and 20 nymphs were collected from cattle control in naturally parasitized bovines, (n =114) and water buffaloes (n = 88) kept pointing out the best cost-benefit. It was by small-holder dairy farmers (<10 animals). selected 72 bovines with superior counting All ticks were identified morphologically to 10 ticks and divided in four experimental to species level using dichotomous keys groups: G1-Fipronil 1mg/kg (TopLine Pour- and then representative specimens were on); G2-Fluazuron 2.5 mg/kg (Forbox Pour genetically characterized using primers on); G3-Moxidectin 1mg/kg (Onix Injectable); targeted at cytochrome c oxidase subunit 1 G4-chlorpyriphos 30g, cipermethrin 15g (COX1), internal transcribed spacer 2 (ITS2) and fenthion 15g (Colosso FC30 - spray). of the nuclear ribosomal DNA and 16S gene. Treatments was realized according to DNA sequences of all three regions were manufacturator reccomendations. The G4 analyzed separately using Bayesian Inference group presented best reduction percentual, and Neighbor Joining phylogenetic methods. greater 7° Post Treatment Day (PTD) indice Results: Morphological and molecular (83.23%) and a 35° PTD without effect. The identification of 753 specimens showed that second best strategy was G3 with inverse they belonged to Hyalomma (Hy.) anatolicum reduction percentage, presenting a worst (n = 680), Hy. hussaini (n = 3), Hy. scupense indice in first weeks and better results in last (n = 1), Rhipicephalus (Rh.) microplus (n treatments (82.85% in 28°PTD and 44.48% in = 66) and Rh. (Boophilus) annulatus (n = 35° PTD). G1 and G2 had a better reduction 1). Phylogenetic analyses of three genetic percentage in 21° PTD (32.63 and 2.79%, markers revealed high diversity in all tick respectively), with insatisfatory percentage species genetically characterized. in other dates. Group one, two, three and four presented a USD 0.37, USD 0.36, USD Conclusion: This study provides a detailed 1.61 and USD 0.28 respectively, for each account of the genetic diversity of hard ticks treatment and each animal. G1, 2 and 3 need

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 211 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

animal weighing and could be measured using 3 samples for Borrelia spp.(0.14%) and 2 balance (fast and high cost) or weighing sample for B.gibsoni(0.09%). thoracic tape (low cost and slow). For G4, a spray chamber was adquired with a USD In this study, molecular surveillance of tick- 4,818.63 cost, making a onerous strategy but, borne rickettsial and protozoan infectious for herds above 5,000 head, the return for diseases was conducted to investigate the low cost of the dose per animal can occur the infectious status in the Republic of in less than a year. In summary, the acaricide Korea. Continuous monitoring of tick-borne contact formulation administered through the infectious diseases is needed to prevent spray chamber presented a better reduction neglected tick-borne zoonoses having percentage, easier application and lower cost potential emerging diseases in the future due per treatment, and could be indicated as a to climate change. strategy of better cost-benefit for medium and large herds. PS01.48 Mortality Time of Rhipicephalus Microplus Larvae Immersed in Binary PS01.47 Molecular Surveillance of Tick-Borne Mixtures of Essential Oils Diseases in Dogs, 2017–2018 Leandro Rodrigues1, Dr. Cecília José Jinhyeong Noh1, Hyunkyoung Lee2, Mi-Sun Veríssimo1, Karen Tieme Akashi3, Dr. Luciana Yoo 1, Yeojin Park1, Subin Min1, Jiyeon Lim1, Morita Katiki1, Dr. Mariana Santos De Miranda1, Bang-Hun Hyun1, Yun Sang Cho1 Poliana Araújo Pacheco2, Dr. Rodrigo Giglioti1 1Parasitic and Insect disease Laboratory, 1Institute of Animal Science, Brazil, 2University Animal and Plant Quarantine Agency, of São Paulo, Brazil, 3State University of Republic of Korea, 2Animal Pathodiagnostic Maringá, Brazil Laboratory, Animal and Plant Quarantine Agency, Republic of Korea Rhipicephalus microplus causes damages to livestock and transmits diseases to its The frequency of canine tick-borne hosts. Essential oils from different plant diseases, which is babesiosis, granulocytic species are toxic to the larval stage of this anaplasmosis, monocytic ehrlichiosis and parasite. The objective of this work was to lyme disease are increasing globally including evaluate in vitro the mortality time of R. Korea. The aim of this study was to identify microplus larvae immersed in combinations tick-borne pathogens in dogs and provide (1:1) of the mixture of ten essential oils base line data in controlling of tick-borne (anethole, vanillin, carvacrol, cinnamaldehyde, diseases that may affect human and animal 1,8-cineole, carvone, eugenol, linalool, thymol health in Korea. and L-limonene) aiming to evaluate possible synergistics effects of these compounds. The Canine blood samples were taken from test consisted of immersing approximately companion dogs in veterinary clinic and shelter 100 larvae of R. microplus in 200 μl of the dogs in abandoned animal shelter. DNA purified binary mixture, putting each one and its from blood samples was used for the detection repetition in one well of a 48 well plate; of 6 of tick-borne diseases (Anaplasma then, with the aid of a magnifying glass and phagocytophilum, A. platys, Ehrlichia a cronometer, we verified the time in which chaffeensis, E. canis, Borrelia spp., Babesia spp.) 100% of the larvae were dead. The ten binary using previously reported PCR assays. mixtures with the shortest mortality time were diluted in soybean oil: 50, 25, 12.5, 6.25, A total of 2,215 dog blood samples were 3.125 and 1.56%. Negative control (soybean collected. Whole bloods’ DNAs were oil, S) and two Pour on acaricides were used extracted from samples using Maxwell as positive controls (C1: Flumethrin 1g, and 16 whole blood DNA kit according to the C2: Cypermethrin 5.0 g + Chlorpyrifos 7.0 g manufacturer’s instuction. As a result of PCR + Citronelal 0.5 g). All binary mixtures, doses analyzed 16s rRNA gene, total of 11 samples and the control treatments were tested in were positive for A. phagocytophilum(0.5%), triplicates. The larvae mortality time data

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 212 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

were analyzed by the PROC GLM procedure, PS01.50 Molecular Characterization of whose model included the fixed effects of the Fasciola Hepatica from Indigenous Animals compound, dose and its interactions, and the in Cajamarca, Peru in Comparison with That means were compared by the tukey test (p from Livestock <0.05). The most toxic mixture for the larvae was thymol + limonene, because at the dose of Dr. Madoka Ichikawa-Seki1, Dr. Pedro Ortiz2, 1.56% presented the shortest mortality time (12 Dr. Cristian Hobán, Dr. María Cabrera, Dr. José min) (p 0.05). Mortality time for the controls Angulo-Tisoc were: C1, 66.67 ± 1.53; C2, 141.0 ± 1.0 minutes; 1Laboratory of Veterinary Parasitology, S, 2400.1±0.1˂ min. It is concluded that this Iwate University, Japan, 2Laboratorio methodology is easy to do, low cost and able de Inmunología, Facultad de Ciencias to test in vitro toxic oil substances for larvae. Veterinarias, Universidad Nacional de Cajamarca, Peru, 3Instituto Veterinario de Investigaciones Tropicales y de Altura, Sede PS01.49 Efficacy of Oral Formulation of Marangani, Universidad Nacional Mayor de Sarolaner (SimparicaTM) Against Naturally San Marcos, Peru Infestation Brown Dog Ticks in Dogs Presented as Veterinary Patients in Thailand Fasciola hepatica is widely distributed in the temperate regions including South American Dr. Piyanan Taweethavonsawat1, Dr. Thitirat countries. It has been speculated that F. Chaimee2, Dr. Patcharin Priyavoravong2, Dr. hepatica was introduced from European Thunchanok Jantanawaranon2, Dr. Chatree countries to South American continent along Khatiworavage1 with movement of animals. However, limited 1Chulalongkorn University, Thailand, 2Zoetis information is available to reveal the actual (Thailand) Limited, Thailand dispersal route. In this study, we focused on Peruvian F. hepatica collected from indigenous Brown dog ticks are recognized as one of animals Cajamarca, Peru. the most prevalent ectoparasites in Thailand. Ticks are vectors for many pathogens that Fasciola flukes collected from 11 alpacas and 15 can cause several diseases in dogs. Treatment guinea pigs in Cajamarca were fixed with 70% and control ectoparasites infestations on dogs ethanol. One or two flukes were analyzed per is an important role for general veterinary animals. Species identification was performed practice. The purpose of this study was by using the DNA fragment patterns of nuclear evaluated the efficacy of oral formulation phosphoenolpyruvate carboxykinase (pepck) of sarolaner treatment naturally infestation and DNA polymerase delta (pold) genes. brown dog ticks in dogs. Sixty dogs were Again, the DNA sequences of mitochondrial selected into study from overall country. NADH dehydrogenase subunit 1 (nad1) gene One dog in each household was allowed were analyzed to reveal the phylogenetic to be enrolled as the primary patient. This relationship with F. hepatica from livestock primary patient had to harbor at least 3 (cattle, sheep, pigs) collected from the attached alive-ticks at enrolment. Dogs same area. The median-joining network was received oral formulation of sarolaner follow constructed to compare the nad1 sequences recommendation dose at Day 0. Tick counts with the reference European countries. on primary dogs were conducted prior to treatment on Day 0, and on post-treatment All of the Fasciola flukes collected from at Day 7 and 30. Dogs were thoroughly alpacas and guinea pigs were identified as F. examined for within 10 minutes by blinded hepatica by using nuclear pepck and pold. study personnel until all ticks were removed. Again, the mitochondrial nad1 sequences of F. Efficacy of sarolaner against ticks was 93%, hepatica from alpacas and guinea pigs were 96.7% on Day 7 and 30 respectively. Sarolaner almost identical to those of F. hepatica from administered orally at monthly intervals at a livestock which were previously reported. recommendation dose was highly effective Therefore, it is demonstrated that there is against naturally infestation brown dog ticks no difference between Peruvian F. hepatica on dogs in Thailand. populations from indigenous animals and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 213 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

livestock. Since Peruvian F. hepatica population control group received no treatment. Fifteen had close relationship with Spanish F. hepatica days later, all the sheep were sacrificed population, it is concluded that F. hepatica was and the number of adult Fasciola parasites introduced from Europe to Peru. present in the livers was counted. The results of the controlled efficacy test in sheep show a negative TCBZ efficacy (-125.7%). It is PS01.51 Controlled Efficacy Test of concluded that cases in which therapeutic Triclabendazole Against Fasciola Hepatica cure did not occur appear to be related to Derived from Human Infections resistance of F. hepatica to TCBZ.

Dr. Pedro Ortiz1, Dr. Marco Rivera-Jacinto2, Dr. Claudia Rodríguez-Ulloa2, Silvia Chilón2, PS01.52 Seasonal Re-emergence of Fatal Cristian Hobán1, María Cabrera1, César Murga- Acute Immature Paramphsitomosis in Goats Moreno1, Dr. Corpus Cerna1 After Flood – A Longitudinal Study in Tamil 1Laboratorio de Inmunología, Facultad de Nadu, India Ciencias Veterinarias, Universidad Nacional de Cajamarca, Cajamarca, Peru 2Laboratorio de Dr. Soundararajan Chinnaiyan1, Dr Nagarajan Microbiología, Facultad de Ciencias de la Salud, Kumaresan2, Dr Arul Prakash Mani3 Universidad Nacional de Cajamarca, Peru 1Tamilnadu Veterinary and Animal Sciences University, India, 2Tamilnadu Veterinary and Cajamarca region of Peru is an area highly Animal Sciences University, India, 3Tamilnadu prevalent of Fascioliasis caused by Fasciola Veterinary and Animal Sciences University, India hepatica. The parasite affects domestic species including man. Currently, human Parampisotomosis causes severe morbidity patients are treated with a human formulation and mortality in ruminants in India. of Triclabendazole (TCBZ) (Egaten®). Outbreak of acute paramphistomosis in The objective of the present work was to goats maintained under semi-intensive at investigate the efficacy of TCBZ in humans Mottur-Siruvakkam village, Kancheepuram infected with the liver fluke F. hepatica district, Tamil Nadu was carried out from using a controlled efficacy test in sheep June 2015 to May 2016. The overall mortality experimentally infected with metacercariae and morbidity was 58.82 and 35.95%. derived from human isolates of F. hepatica. Among the seasons, highest mortality was Twenty-six patients were treated with TCBZ, observed during winter (50.67%) followed divided in two groups. Patients in group A by summer (13.51%), northeast monsoon (n = 13) were treated with TCBZ at 10 mg/ (2.67%) and no mortality during south west kg bodyweight and those in group B (n = 13) monsoon. Similarly, highest morbidity was with 20 mg/kg. Oral treatment with TCBZ was observed only during winter (336.67%) and administered on two consecutive days. Five no morbidity was observed during other patients from both groups failed to cure after seasons. Incubation period was 2 days in TCBZ treatment. From those patients that goats. Adult paramphistomes observed in did not cure, F. hepatica eggs were collected rumen were Cotylophoron cotylophorum and the miracidia produced was used to (40.54%), Gastrothylax crumenifer (29.19%), infect Lymnaea snails. Sufficient amount Paramphsitomum cervi (19.19%), Fischoederius of metacercariae was obtained to perform elongatus (8.38%) and F.cobboldi (2.70%). a controlled efficacy test in sheep. Twelve Affected animals showed severe diarrhoea sheep were artificially infected orally, with two with bad smell, bottle jaw, protrusion of hundred metacercariae each, derived from mucous membrane of eyes and died with human patients. The infected sheep were 2 days after recumbent. On postmortem randomly divided into two groups: treated examination, adult paramphistomes were group (n = 6) and control group (n = 6). 106 found in rumen and mmature paramphistomes days after infection, animals in the treated were found mostly in the duodenum and group were treated by oral administration abomasum. Gall bladder and urinary bladder of TCBZ (Bilevon®) at the dose of 10 mg/kg were enlarged and nodules found on the live weight as a single dose. Animals in the spleen. On histopathology, rumen showed

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 214 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

presence of adult paramphistome and few enough to avoid disease. Knowledge presence of plug of ruminal papillae in the oral of actual viability of oocysts is therefore cavity of adult paramphistomes. In immature required for determination of optimal dosage. paramphistomosis, stunting of intestinal villi, Unfortunately, no rapid and accurate method desquamation of intestinal villi around the for determination of oocyst viability exists. worm, necrosis of villi, infiltration of MNC Difficulty assessing oocyst viability ultimately (eosinophils, lymphocytes, neutrophils), impedes successful implementation of glandular hyperplasia were observed. There vaccination programs. We have demonstrated was presence of immature paramphistomes an in vitro assay that uses transcriptional deeply embedded in the submucosa as well activity as a proxy for viability of Eimeria as just above the Brunner’s glands. Snails spp. oocyst viability. We have investigated collected from the lake were identified as a range of specific assay targets, with early Indoplanorbis exustus (79.54%) followed by data showing a strong correlation between least number of Pila globosa (12.50%), Pila abundance of these targets and oocyst virians (3.41%), Bellamya b.eburnea (3.41%) viability. Assay optimization of transcript and Stenothyroides blandfordiana (1.14%). quantification protocols, including assessment Amphistome cercariae were recovered from of one- and two-step RT-qPCR strategies and I.exustus snails. the use of digital droplet PCR (ddPCR) aim to increase sensitivity. Other potential assay applications include use as an epidemiological PS01.53 Transcriptional Activity as a Proxy tracking tool or validation of strategies for for Eimeria Spp. Oocyst Viability: The environmental control of Eimeria and related Key to Optimizing Coccidiosis Vaccine parasites. Performance?

Perryn Kruth1, John R. Barta1, Ryan Snyder1 PS01.54 Efficacy of Three Avermectins 1University Of Guelph, Canada in EPRINEX®, IVOMEC® F and DUOTIN® Formulations Against Dermatobia Hominis Eimeria spp. (Apicomplexa) infect the Larvae in Cattle Naturally Infested in Brazil digestive tract of a wide range of vertebrates. These parasites are generally of little concern Debora Sartor1, T Azevedo2, Cristiano in wildlife, companion animals, and humans. Nascimento3, Luis Pelissoni1, Gustavo Sabatini1, In the commercial poultry industry, where Fabio Scott2, Andre Cutolo1 animal stocking densities are high and feed- 1Boehringer Ingelheim Animal Health Do Brasil conversion efficiency is of top concern, they Ltda., Brazil, 2Universidade Federal Rural do are a major biological threat. Oocysts are Rio de Janeiro, Brazil, 3Convolution, Brazil shed in feces of infected birds, and sporulate to become infective to poultry. Parasite Avermectins (abamectin, ivermectin and replication damages enterocytes to produce eprinomectin) are veterinary endectocides disease ranging in severity from simple used on livestock having a broad-spectrum unthriftiness to death. Severity of this self- of activity and a safe profile. Dermatobia limiting infection is a function of three major hominis is amongst the main ectoparasites of factors: Number of infective oocysts ingested; bovines, being widely distributed in tropical parasite species; and the host’s previous and subtropical regions of Latin America. exposure to that species (i.e. immune status). Economic losses caused by this parasite in Infection protects the host against future Brazilian herd were estimated at US$ 380 infection by homologous species; several million per year. The aim of the study was rounds of infection produce sterile immunity. to evaluate the efficacy of eprinomectin in The self-limiting and immunogenic nature of EPRINEX®, ivermectin in IVOMEC® F and Eimeria spp. make live vaccination an ideal abamectin in DUOTIN® against Dermatobia strategy for coccidiosis control. Vaccination hominis. 40 bovines naturally infested with requires establishment of infection with a minimum of 10 grubs per animal were sufficiently numerous viable oocysts to used and splitted in four groups with 10 stimulate development of immunity, but animals each: EPRINEX® Pour On (500

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 215 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

g of eprinomectin/kg of body weight), were tested on the egg hatch inhibition assay IVOMEC® F (200 g of ivermectin/kg of (EHIA) and the larval development inhibitions bodyµ weight + 2 mg of clorsulon/kg of body assay (LDIA) and compared with Albendazole weight, subcutaneousµ injection), DUOTIN® the positive control. The concentrations for (200 g of abamectin/kg of body weight, the plant extract, fractions and Albendazole subcutaneous injection) and an untreated used for the study were 0.78, 1.56, 3.125, 6.25 group.µ The products were administered and 12.5mg/ml. The results showed that the to the study animals in the treated groups crude extracts, fractions and Albendazole at following the manufacturer’s instructions. concentration dose of 12.5 mg/ml produced The animals were examined visually and by 100% inhibition of egg hatching and larval palpating the back and the sides of each development of Haemonchus contortus. animal for the presence of live subcutaneous Although, there was no significant difference grubs before and after treatment to calculate (p>0.05) in the mean percentage egg hatch the efficacy. The number of nodules with and larval development inhibition of the live larvae was counted 7 and 14 days after crude extract and fractions when compared treatment to evaluate efficacy. The tested with Albendazole. However, significant products were effective in controlling grubs difference (p<0.05) was observed with from all treated groups. The percentages of n-butanol fraction which inhibited 96.17% of efficacy (arithmetic means) on days 7 and egg hatchability. The extract, fractions and 14 respectively for EPRINEX® were 92.25% Albendazole showed ovicidal and larvicidal and 92.18%; for IVOMEC® F were 93.02% and activity and produced over 50% inhibition 94.90% and for DUOTIN® were 90.70% and of egg hatching and mortality of larvae at 92.52%.These data showed that EPRINEX®, concentration ranges of 0.78 - 12.5 mg/ml. IVOMEC® F and DUOTIN® were effective in the The result obtained from this study suggests treatment of D. hominis grubs in cattle. No that H. pachyanta possess anthelmintic adverse reactions were observed in any of the effects against H. Contortus and validates the treated animals. folkloric use of the plant in the management of H. Contortus infections.

PS01.55 In Vitro Anthelmintic Activity of Hymenodictyon pachyanta Stem Bark Extract PS01.56 Anthelmintic Activity of Icacina and Fractions Against Haemonchus contortus Trichanta Extract and Fractions Against Haemonchus Contortus Prof. Olorunfemi Jegede1, Dr. Olutayo Olayemi1, Dr. Michael Onakpa1 Dr. Monday Michael Onakpa1, Dr. Olutayo- 1University Of Abuja, Nigeria Daniels Olayemi2, Dr. Victoria Elochukwu2, Prof. Cornelius Olorunfemi Jegede2 The developoment of helminthe resistance 1Department of Pharmacology and Toxicology, and high cost of synthetic anthelmintics has Faculty of Veterinary Medicine, University of prompted the need for an in vitro anthelmintic Abuja, Nigeria, 2Department of Parasitology evaluation of crude stem bark extract and and Entomology, Faculty of Veterinary fractions of Hymenodictyon pachyanta Medicine, University of Abuja, Nigeria plant as an alternative in the management of endoparasites including Haemonchus Haemonchus contortus is an important contortus which is one of the most prevalent gastrointestinal parasite of ruminants, which parasitic nematodes in small ruminant farming causes anaemia, submandibular oedema, globally. Hymenodictyon pachyanta stem bark progressive weight loss, weakness and even is used as anthelmintic by indigenous farmers death. This study was aimed at evaluating in Nsukka, Enugu State and Gwagwalada, the in vitro anthelmintic activity of Icacina Federal Capital Territory, Nigeria. The stem trichanta leaf extract and fractions against bark of H. pachyanta were collected from H. Contortus eggs and larvae. Phytochemical the field in Nsukka, Enugu State, air dried, analysis of the crude extract was carried pulverized and extracted with 80% Methanol. out using standard techniquesto detect The extract and fractions of H. pachyanta secondary metabolites contained in the plant.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 216 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

The in vitro anthelmintic activity of the crude demarcated on the lawn next to the Biological extract and fractions was determined using Control Laboratory of the Advanced Research eggs hatch inhibition assay (EHA) and larval Center in Campinas, State of São Paulo, development inhibition assay (LDA), while Brazil, where a small group of capybaras Albendazole and distilled water were used as (Hydrochoerus hydrochaeris) maintained a positive and negative controls respectively. population of Amblyomma ticks. Each set was The extract of I. trinchanta leaves inhibited assigned to treatment with either Beauveria hatching of eggs and larval development of bassiana IBCB 66 or Metharhizium anisopliae H. contortus in a concentration-dependent IBCB 425 from the Oldemar Cardim Abreu manner. Concentrations of 0.78, 1.56, Entomopathogenic Funghi Collection of the 3.12, 6.25 and 12.5 mg/ml of the extracts Biological Institute. Emulsions containing completely inhibited the hatching of eggs infectious propagules of each strain were and development of larvae and these were sprayed on the respective parcels at 5×10¹¹, comparable to Albendazole anthelmintic 5×10¹² and 5×10¹³ conidia/ha. Control parcels activity. At the concentration of 12.5 mg/ were sprayed with the same volume of water. ml, the crude extracts, and fractions and Ticks were recovered from the lawn parcels Albendazole produced 100% ovicidal and with the aid of dry ice traps after application larvicidal activity against egg hatching and and then at four consecutive weekly intervals. larval development inhibition of H. Contortus, Six traps were laid concomitantly on each except for n-butanol and ethyl acetate parcel. Because of the season, only nymphs fractions which inhibited 98% and 99.8% and adults were captured. Ticks were counted, of egg hatching and larval development identified and placed in test tubes closed with respectively. However, there was no significant cotton stoppers, corresponding to each single difference (p>0.05) in the anthelmintic trap. Tubes were kept in an incubator at 28°C activity of the crude extracts and fractions and 80% humidity. Mortality was checked +1, when compared to the anthelmintic effects +3 and +10 days after capture. Provided the observed with Albendazole activity. The result number of ticks captured in each trap in each in this study showed that crude extract and occasion was random and eventually zero, the all the fractions tested possess anthelmintic proportion of dead ticks was used to enable compounds, warranting further in vivo comparison of the different treatments. evaluationfor its safety and toxicity profiles. Mortality and treatment were significantly associated in all cases (p < 0.001). Correlation coefficients for Beauveria and Metharizium PS01.57 Dose Determination Study were -0.674 and -0.466 for nymphs and of Entomopathogenic Funghi for the -0.619 and -0.589 for adults, respectively. Environmental Control of Amblyomma Ticks The best LC adjustment was obtained with Cauchy’s generalized linear model for binary Shirley Batista da Silva Araújo1,4, Karina data. Median₅₀ lethal doses for Beauveria and Araújo dos Anjos1,4, Fernanda Calvo Duarte2, Metharizium were 1.69×10¹² and 3.82×10¹² Leonardo Costa Fiorini2, Paulo Henrique conidia/ha for nymphs and 8.19×10¹² and Selbmann Sampaio2, José Eduardo 5.93×10¹² conidia/ha for adults, respectively. Marcondes de Almeida3, Márcia Cristina The current results will support further Mendes2 research focused on the biological control of 1Master’s Program in Health, Food and ticks on lawns and pastures. Environmental Safety in Agribusiness, Biological Institute, Brazil, 2Animal Health Research Center, Biological Institute, Brazil, 3Plant Protection and Animal Health Advanced Research Center, Biological Institute, Brazil, 4Coordination of Superior Level Staff Improvement (CAPES) scholarship, Finance Code 001, Brazil

Two sets of four 6 m square parcels were

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 217 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS01.58 Foldscope as a Diagnostic Tool diseases. Authors are thankful to Department for Identification of Canine Parasites with of Biotechnology, Government of India for Emphasis on Zoonotic Parasites providing the financial grant and facilities to carry out the research work. Dr. Mandeep Singh Bal1, Dr. Rashmi Sagar1, Dr. Paramjit Kaur1, Dr. Vishal Mahajan1, Dr. Gursimran Filia1 PS01.59 Efficacy of Oral Afoxolaner 1Guru Angad Dev Veterinary & Animal (NexGard®) and Afoxolaner Plus Milbemycin Sciences University, India (NexGard Spectra®) Against Rhipicephalus Sanguineus and Ctenocephalides Sp. In The study was undertaken to use foldscope Naturally Infested Dogs in Brazil. (magnification 140x) as a portable diagnostic tool for canine parasitic infections. Foldscope Andre Cutolo1, Luis Pelissoni1, Debora Sartor1, is a paper microscope developed in USA & is Gabriela Vito1, Cristiano Nascimento2, Tassia attachable with smart phone to visualize and Vale3, Isabella Souza3, Naylene Silva3, Livio capture pictures of specimens/slides. Faecal Costa Junior3, Maycon Heidmann4, Itallo samples were collected from 173 dogs (83 pet, Carvalho4, Marcela Mendes4, Taminis Santos4, 90 stray) from different districts of Central Isabella Chagas4, Bruno de Castro4 plain zone of Punjab, India. Skin scrapings 1Boehringer Ingelheim Animal Health Do were also collected from dogs (15) with Brasil Ltda., Brazil, 2Convolution, Brazil, history of dermatitis and ticks from dogs (10) 3Universidade Federal do Maranhão, Brazil, with tick infestation. Samples were processed 4Universidade Federal do Mato Grosso, Brazil as per standard parasitological techniques and examined by foldscope and optical Rhipicephalus sanguineus ticks and microscope. Ctenocephalides felis fleas are the most prevalent ectoparasites of dogs in Brazilian Overall 86 faecal samples (49.71%) were urban areas, potentially causing irritation, found positive for gastrointestinal parasites blood depletion, pruritus, skin lesions and with predominance of hook worm eggs pathogen transmission. Afoxolaner is a (46.24%), followed by Taenid eggs (1.73%), molecule from the isoxazoline family with coccidian oocysts (1.15%) and Toxocara eggs proven ectoparasiticide action against fleas (0.58%). Ticks collected were identified as and ticks infesting dogs. Milbemycin Oxime Rhipicephalus sanguineus. Skin scrapings is an antiparasitic endectocide belonging were found positive (26.67%) for Demodex to the group of macrocyclic lactones and is spp. mites. Gastrointestinal parasitic infections active against several gastrointestinal worms. were found to be higher in stray dogs Afoxolaner (NexGard®) and Afoxolaner + (83.33%) than pet dogs (13.25%). Low grade Milbemycin Oxime (NexGard Spectra®) were gastrointestinal parasitic infections were administered to the study dogs per the label not detected by foldscope. Relative efficacy instructions. Different breed owned dogs, of foldscope in detecting ticks, mites and heavier than 2kg and older than 8 weeks parasitic eggs/oocysts evaluated as 100%, old, from urban areas of São Luis, Maranhão 80% and 75.6%, respectively as compare to state, Sinop and Ipiranga do Norte, Mato conventional light microscope (assuming Grosso state, were visited in their houses and efficacy of light microscope as 100%). included in the study if harboring more than Examination of parasitic stages in faecal and 6 fleas and/or more than 4 live attached ticks. skin scrapings by foldscope showed problem 136 tick infested dogs and 146 flea infested in focusing of glass slides. Therefore, a new ones were included. Part of the animals method using paper slides with slits was also harboured mixed infestations. Dogs from a used in the study. Presence of parasites viz; same house were treated with the same test Hook worms, Toxocara spp. and Taenid eggs product. Sixty-seven dogs infested with ticks in dogs indicate possible risk of zoonotic and 73 dogs infested with fleas received infections to the people of the region. NexGard® and 69 dogs with ticks and 73 dogs Foldscope may be used as a novel, low cost with fleas received NexGard Spectra® single tool for quick diagnosis of canine parasitic oral dose at Day 0. Efficacy was calculated

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 218 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

based on the comparison of the arithmetic compared to ELISA. Positive samples with mean number of ticks or fleas before treatment coproscopy confirm the presence of Giardia versus the mean number at Day 30 (±4). The cysts, however negative results cannot mean number of ticks pre-treatment and exclude the presence of the parasite. In this percentage of efficacy were respectively for study, 10 negative samples with coproscopy the NexGard® and NexGard Spectra® groups presented specific Giardia antigens with the 21.3 and 98.5%; 31.1 and 98.3%; and for fleas ELISA test. respectively 46.4 and 99.2%; 50.9 and 99.8%. Both NexGard® and NexGard Spectra® showed The combination of both coproscopy and excellent efficacy results against R. sanguineus soluble antigen detection increases the ticks and Ctenocephalides sp. fleas in housed probability of correctly identifying negative naturally infested dogs in Brazil. samples. In comparison with combined coproscopy and ELISA, Speed™ Giardia presented good sensitivity (87.9%) and PS01.60 Efficacy of the Rapid In-Clinic specificity (95.7%). Speed™ Giardia Test for the Detection of Giardia Duodenalis in Dog Faeces Compared Conclusion: This study confirms the strong to ELISA and Coproscopy agreement between the Speed™ Giardia test and ELISA for the detection of Giardia Dr. Stéphane Audhuy1, Dr. Sofia Loukeri2, Dr. antigens from faecal samples. Speed™ Giardia Coralie Ségalen2, Dr. Christelle Navarro2, Dr. can be an efficient way for identification Emmanuel Claret1, Dr. Vanessa Chala2 of canine Giardia infections in-clinic. A 1Research & Development, BVT-Virbac, France, combination of coproscopy with an antigen 2Global Medical Department, Virbac, France detection test increases the probability of identifying canine giardiasis. Giardia infection is a frequent cause of diarrhoea in dogs and giardiasis should be added on the differentials for any dog with PS01.61 The AAVP Educators Meeting: Past, acute or chronic diarrhoea. The aim of the Present, and Future study was to evaluate the diagnostic efficacy of the rapid in-clinic immunochromatographic Prof. Dwight Bowman1, Prof. Heather Walden2, test, Speed™ Giardia (Virbac) and to compare Prof. Brian Herrin3, Prof. Byron Blagburn4 the accuracy of the immunochromatographic 1CVM Cornell University, United States, test and the zinc sulfate flotation technique 2University of Florida CVM, United States, (coproscopy) versus the ELISA, ProSpecT™ 3Kansas State University CVM, United States, Giardia EZ Microplate Assay (Thermo Fisher 4Auburn University CVM, United States Scientific). The first AAVP Educators Meeting occurred Method: Feacal specimens from 104 dogs were in 2006 in Atlanta, GA, with a $15,000 first examined with coproscopy. Then, samples donation from the Companion Animal were blindly tested with Speed™ Giardia test Parasite Council [CAPC] to cover meals and and ProSpecT™ Giardia assays which served as food and AAVP supplying up to $300 travel the gold standard for this study. funds for one AAVP faculty member from each North American Veterinary School or Results: In comparison with the gold standard, College. Additional faculty could attend Speed™ Giardia demonstrated high sensitivity at their own expense. The CAPC donation (92.7%, CI95 of 82.7% to 97.14%), specificity has continued allowing the meetings to (95.9%, CI95 of 86.3% to 98.9%) and positive occur biannually. Since 2006, meetings and negative predictive values of 96.2% and have followed at the CVM Oklahoma 92.1% respectively. The results obtained with State University, Stillwater (2009); USDA- the two soluble antigen detection methods ARS, Beltsville, MD (2011); Loews Hotel, presented a strong agreement (94.2%). Chicago IL (2013); AVMA headquarters, Coproscopy presented a very good specificity Schaumburg, IL (2015), the CVM Ohio State (100%) but lower sensitivity (81.8%) when University, Columbus (2017), and coming

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 219 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

in 2019 at the CVM University of Florida in Zambia, 6Institute of Parasitology, McGill Gainesville. The meeting format, including University, Canada, 7Brantam House, South plenary speakers and group exercises Africa, 8Virginia-Maryland Regional College for analysis and discussion, has provided of Veterinary Medicine, Population Health interactive peer support to instructors in Sciences, United States veterinary parasitology worldwide, provides opportunities for exchange of information on Since 1997, the World Association for the teaching strategies, course content, computer Advancement of Veterinary Parasitology hardware and software platforms, specimens (WAAVP) African Foundation (AF) has for laboratories, suggestions for examination achieved a “first“ by establishing a successful content and format, and information on endowment which awards travel scholarships the accreditation process. The Symposium to Next Generation (NG) African veterinary aids instructors in establishing core content parasitologists to present their research for NAVLE preparation and has generated findings at WAAVP Biennial Conferences. To the publication: AAVP recommendations date, 200 NG veterinary parasitologists from for core competency standards relating to 24 African countries submitted applications; parasitological knowledge and skills. Snowden 80 from 15 countries were awarded et al., JVME 43:344-8;2016. This meeting scholarships to present research at WAAVP is especially important for parasitologists conferences in Europe, Asia, South America, entering the field and for new and veteran USA, Canada, Australia and New Zealand. faculty who are developing resources for courses and laboratories, for the sharing The WAAVP AF goals are to support deserving of ideas, networking, the development of awardees ready to present their research to avenues for teaching advice, and sharing an international audience, to develop global of resources (i.e. specimens, photos, case networks, and to bring positive recognition reports, etc.). Participant feedback supports to their institutions, countries and academic the value and need for continuation of the fields. The WAAVP AF Committee oversees a program and recovgnizes its value in when successful financial investment and biennially questioned as to the importance of the invites approximately 100 global stakeholders material within curricular discussions within to assist with recruitment across Africa. universities. This biannual meeting has become a valuable part of AAVP and our From 2008-2017, 42 awardees participated continued efforts to strengthen and expand in these conferences, and 37 submitted veterinary parasitology education. narratives which were published in the WAAVP quarterly online newsletters (https:// www.waavp.org). These narratives were PS01.62 A Model of Success: World analyzed using QDA Miner 5.0 (Provalis Association for the Advancement of Research, Montreal, QC, Canada). The coding Veterinary Parasitology African Foundation process started with an initial open-minded (1997-2017) reading to learn the data. During the second reading, each narrative was scrutinized for Prof. Rosina C. Krecek1, Ms. Maggie Fisher2, emerging mutually exclusive categories or Prof. Annemarie Avenant-Oldewage3, Prof. themes. A third read through was performed Banie L. Penzhorn4, Prof. Isaac K. Phiri5, Prof. after coding to fine-tune the results. Roger Prichard6, Mr. Derek Sumption7, Prof. Stephen R. Werre8 Nine themes emerged from the analyses 1Texas A and M University, United States, with the top five being: 84% expressing 2Shernacre Enterprise Ltd., The Mews Studio, gratitude to WAAVP AF for their conference Portland Road, United Kingdom, 3Department opportunity, 76% for positive learning of Zoology, University of Johannesburg, South experiences, 60% for exposure to current Africa , 4Department of Veterinary Tropical research, 51% for networking opportunities Diseases, Faculty of Veterinary Science, and 35% for interaction with other scientists. University of Pretoria, South Africa, 5School The WAAVP AF promises to continue for the of Veterinary Medicine, University of Zambia, foreseeable future. It attributes 20 years of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 220 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

success to being financially well invested in the importance of heartworm disease, their South Africa, a dedicated Committee and future role in prevention of this disease, awardees committed to identify innovative and the significance of offering education and novel solutions to address parasites that to support prevention awareness. Further plague animal and human health. community and academic teaching and learning opportunities, which support these educational needs are warranted. PS01.63 Development of Veterinary Students’ Knowledge, Attitudes, and Practices Towards Dirofilaria Immitis in St. Kitts PS01.64 Serological Evidence of Canine Arthropod-Borne Infections in an Ecotone Dr. Mary Mauldin Pereira1, Ms. Kaitlyn Jonk1, Area of a Natural Reserve at the Pantanal, Dr. Jennifer Ketzis1, Dr. Anne Conan1 Brazil 1Ross University School of Veterinary Medicine, St. Kitts Bruno Alberigi1, Celeste Souza2, Flavia Cardoso2, Caroline Magalhães2, Caroline Dirofilaria immitis (heartworm) has a spatial Almeida2, Prof. Flavya Mendes-de-Almeida3, heterogeneous distribution on the Caribbean Prof. Norma Labarthe4 island of St. Kitts with a documented 1Universidade Federal Rural do Rio de Janeiro, higher prevalence in the southeastern Brazil, 2Laboratório de Imunomodulação region. Numerous Ross University School e Protozoologia, Instituto Oswaldo Cruz, of Veterinary Medicine (RUSVM) students Fundação Oswaldo Cruz, Brazil, 3Universidade reside in this area, and we hypothesize that Federal Fluminense, Brazil, 4Fundação this student population will promote a control Oswaldo Cruz, Brazil campaign for heartworm. To investigate, a standardized questionnaire was developed Arthropod-borne infections are highly to assess RUSVM students’ knowledge, dependent on environmental conditions; attitudes, and practices towards heartworm. therefore, anthropomorphic meddling close Seventy-three incoming students completed to natural reserves may disrupt the natural the questionnaire in September 2017, and balance that maintains wildlife and onehealth. one year later, 38 (52%) of these students Since human populations keep domestic completed the same questionnaire. Majority animals as pets or working animals, it is of respondents were women (~80%) originally common to find dogs roaming in Brazilian from the United States (70 from 31 states). natural reserves and also horses or bovines While there were only five students (7%) working under humane orders. This close living in the southeastern region of the island living favors the spillover of bioagents at the time of the first questionnaire, a 48% that can affect wildlife, domestic animals increase was reported a year later. Student and humans. Dirofilaria immitis, Ehrlichia knowledge surrounding the ability to identify canis, Ehrlichia ewingii, Anaplasma platys, the scientific name of the parasite increased Anaplasma phagocytophilum, Borrelia by 47% in a year’s time. Attitudes toward burgdorferi and Leishmania spp. infect heartworm prevention did not change over unprotected animals without propensity. the year (Wilcoxon paired test, p>0.05), The present survey was conducted using 84 except more advised their family or friends sera from dogs residing at the border area who have a cat in the United States to give of the SESC-Pantanal reserve (16°40´51´´S; it preventative against heartworm (p=0.01). 56º17´45´´W) stored at the Laboratório de Only six incoming students (8%) had a dog on Protozoologia e Imunomodulação, Instituto island versus 15 (39%) after a year. All student Oswaldo Cruz. Samples were tested with dog owners used heartworm preventative, SNAP® Canine Leishmania Antibody Test yet only six used a way to protect their dogs (IDEXX Laboratories) or DPP® canine visceral against mosquitoes. Our results indicate that leishmaniasis test (Bio-Manguinhos) for the despite an improvement of the knowledge, presence of L. infantum antibodies and with attitudes did not change. This highlights the SNAP® 4Dx Plus Test (IDEXX Laboratories) need for veterinary students to understand for D. immitis, Ehrlichia spp., Anaplasma spp.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 221 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

and B. burgdorferi seroprevalences. Ehrlichia cases changed from more than 20 in 2005 spp. seroprevalence was the most frequent to less than 10 in this study. Globally the (76.2%) and of these 19 samples were also average number of annual cases per clinic Anaplasma spp. positive and another sample was reduced when compared to 2005 (44 was positive for the three tick-borne parasites. areas). These results indicate a regression The frequency of Anaplasma spp. positive of the impact of the disease in many parts samples was 27.4%. L. infantum prevalence of France. However, during the recent was 13.1% and D. immitis 7.1%. D. immitis period (2012-20017) the evolution is mostly prevalence was higher than expected in the considered unchanged with an increase in area, although lower than the Brazilian known 8 and decrease in 12 areas. Interestingly in prevalence (23.1%). Tick-borne diseases 12 areas, cases initially considered imported presented high seroprevalence similar to the are now autochthonous. Four of these zones expected for the Pantanal (70.9%), suggesting are located in the Southeast were the vector anthropomorphic impact. L. infantum of Babesia canis is absent. This may suggest prevalence was lower than the known that in these areas where Rhipicephalus prevalence for the Pantanal area (61.7%) sanguineus (s.l) is common, the likely agent suggesting that the biodiversity of the reserve is Babesia vogeli, known quite recently depurates Leishmania circulation enhancing described in France. both wildlife and domestic animal welfare.

PS01.66 Asymptomatic Carrier Status PS01.65 Distribution and Dynamics of and Seroprevalence of Leptospira spp. in Canine Babesiosis in France: Results of a Unvaccinated Canines Presented for Elective Questionnaire Based Survey Sterilization to the St. George’s University, School of Veterinary Medicine Junior Surgery Prof. Patrick Bourdeau1, Florian Carrez1, and Anesthesia Lab Floriane Hubert1, Laetitia Imparato1 1Unit -DPM Veterinary School of Nantes- Rebecca Howell1, Noemi Prieto1, Sarah Oniris, France Stowe1, Norma Valdez1, Elizabeth Wilson1, Dr. Marta Lanza Perea1, Dr. Tara Paterson1, Canine Babesiosis (Babesia canis) is an Dr. Christian Leutenegger2, Dr. Ramaswamy important disease in dogs in France, Chandrashekar2 transmitted by Dermacentor reticulatus. A 1St. George’s University, Grenada, 2IDEXX national survey was conducted in summer Laboratories, Inc, United States 2017 covering the period 2012-2017. Questionnaires were sent to veterinary clinics, The Leptospira genus of bacterium are and distribution and evolution of babesiosis of global concern as pathogenic species was evaluated by using the number of cases cause Leptospirosis – a zoonotic disease diagnosed per year (<1, 1-10, 11-20, 21-50, most prevalent in tropical and subtropical >50), autochthonous (= acquired within the developing countries. Domestic and wildlife territory of activity of the veterinary clinic) or reservoir hosts harbor the bacteria within not. The results were compared to a similar the renal tubules and intermittently shed study conducted in 2005. Distribution was the organism in urine. These asymptomatic analyzed using administrative territorial areas carriers are of significant animal and public (“départements” n= 90). The questionnaires health concern. While the majority of from 620 clinics covered 85/90 areas. The research on the prevalence of Leptospirosis average numbers per clinic was: <1 in 12 areas in animals is based on serology, the few (mainly in South-east and Corsica), 1-10 in 33 studies that have assessed for asymptomatic (West, North, South-east). Higher average carriers within canine populations reported prevalence were detected: 11-20 in 25; 21- prevalence of 0.2 - 22%. There is currently 50 in 13 (versus 32 in 2005) and > 50 cases no data regarding the prevalence of canine in 8 (versus 26 in 2005), mostly distributed asymptomatic carriers of Leptospira spp. in in the Southeast, centre and East of the the Caribbean. The aim of this investigation country. In 14 areas the average number of is to determine the exposure of dogs to

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 222 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Leptospira bacteria using serology and prevalence of Eimeria was 75.5% (1006/1333), assess for the presence of asymptomatic with no difference observed between age carriers using PCR analysis of urine. Using categories, although greater number of oocysts the IDEXX SNAP® Lepto Test, seroprevalence were shed in the lower age groups at 6 months was determined to be 12.65% (21/166) in of age. Thirteen species of Eimeria were 166 owned dogs, not previously vaccinated identified, among which E. bovis, E auburnensis, against Leptospira, presenting to the Junior and E. zuernii were the most predominant Surgery and Anesthesia Lab at St. George’s species in 33.5%, 12.5%, and 11.9% of the positive University School of Veterinary Medicine samples examined, respectively. These were during fall of 2017. Urine samples from 149 of followed by E. pellita (7.4%), E. ellipsoidalis the 166 dogs were tested using a Leptospira (5.4%), E. canadensis (5.3%), E. wyomingensis spp. real-time PCR with 6.71% (10/149) (5.3%), E. bukidnonensis (5.3%), E. brasilensis positive for Leptospira spp. DNA. Nine of (3.4%), E. alabamensis (3.1%), E. subspherica the 10 PCR-positive dogs were followed up (2.9%), E. cylindrical (1.6%), and E. illinoinensis one year later. Blood and urine PCR were (0.8%). There were 358/1333 (26.9%) samples negative in all cases and 55.56% (5/9) were with single species infections and 352/1333 seropositive on SNAP® Lepto Test. Four (26.5%) with mixed infections by two or more of the 9 dogs (44%) were seropositive on Eimeria spp. This is the first large scale study MAT, all of whom had antibodies against reporting on prevalence and diversity of the L. icterohaemorrhagiae serovar, and one Eimeria species in calves throughout Colombia individual also had antibodies against L. and it can be assumed that infections are autumnalis and L. bratislava. The identification ubiquitous in the country and may be playing of Leptospira spp. DNA in canine urine an important role as a subclinical disease samples in this study suggests that there affecting production parameters in conventional are asymptomatic carriers within the canine management systems. population in Grenada at rates similar to that reported in other countries. PS01.68 A Survey of Gastrointestinal Helminths and Coccidia in Llamas and PS01.67 Epidemiological Survey of Eimeria Alpacas Raised in France Species in Naturally Infected Calves Throughout Colombia Ms. Juliette Loiseaux1, Ms. Chloé Defives1, Bruno Polack1, Dr. Jacques Guillot1, Radu Sara Lopez Osorio1, Dr. David Villar-Argaiz1, Blaga1, Mrs. Radia Guechi1, Dr. Veronica Risco- Dr. Klaus Failing2, Dr. Habil Anja Taubert2, Castillo1 Dr. Habil Carlos Hermosilla2, Dr. Jenny J 1Ecole Nationale Vétérinaire d’Alfort (EnvA), Chaparro-Gutierrez1 France 1CIBAV Research Group. University of Antioquia, Colombia, 2Institute for Despite relentless increase of llama (Lama Parasitology, Justus Liebig University, glama) and alpaca (Vicugna pacos) rearing in Germany France, information regarding the presence and prevalence of gastrointestinal parasites There is a lack of information on the in these species remains scarce. This study epidemiology of eimeriosis in naturally infected aimed to determine the presence of helminths cattle in Colombia. A cross-sectional study was and coccidia in llamas and alpacas raised in conducted to evaluate the prevalence, level of France and to identify epidemiological factors infection, and distribution of Eimeria species associated to their detection. The study was in 55 randomly distributed cattle farms across performed between May and July 2017 and 7 States of Colombia. Fecal samples from it included faecal samples from 111 alpacas 1333 calves less than 1-year-old were examined and 150 llamas coming from 22 French farms at a single sampling time from August 2016 located all over the country. All samples were to December 2016. Oocysts were identified analysed by using saturated magnesium to species level based on morphological and sulfate and quantitative McMaster’s technique. morphometric characteristics. The overall Epidemiological analysis to identify risk

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 223 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

factors comprised variables such as camelid incidence across the United States. Given species, sex, age, geographical origin, farming the shared environment in which people and practices (one-species or mixed-species dogs interact, dogs can be both primary farms) and parasite control protocols. hosts and sentinel animals for tick and tick- borne disease distributions for humans. Sampling coverage reached 100% among Therefore, measures taken to better track, farms raising between 2 and 24 individuals prevent, and/or treat tick-borne diseases in (18 farms), and 4 farms with more than 30 dogs can lead to better prevention or control animals sampled between 25% and 37% of of human exposure to these infections. Our their population. Sixteen out of 22 farms study sought to identify demographic and (72%) raised their llamas and/or alpacas biological risk factors for canine seropositivity without contact with other livestock. Twelve to Anaplasma spp., Babesia spp., Ehrlichia farms raised llamas and alpacas together spp., and B. burgdorferi in a cohort of highly (54%). Age varied from one-year old to more exposed hunting dogs in the United States. than 10-years old, while 46% and 31% of Results: After controlling for age and samples were coming from 3-10 years-old and Anaplasma spp. seroprevalence, dogs in less than 3-years old animals, respectively. regions outside of the west were 2.3x more Strongyle-type eggs, Nematodirus sp eggs, likely (90% CI: 1.1274 – 4.5438, p=0.0216) to and Trichuris sp. eggs were found on 19.9%, test seropositive for B. burgdorferi. Dogs 1.9% and 2.7% of animals, respectively. who tested seropositive for Anaplasma spp. Oocysts of Eimeria macusaniensis and small were 1.4x more likely (90% CI: 1.0412 – 1.7846, coccidia (E. punoensis and/or E. alpacae) p=0.0242) to test seropositive for Babesia were found in 8.4% and 75.3% of animals, spp.. Analysis of Babesia spp. seroprevalence respectively. While most results show a low indicated that dogs seropositive for Babesia parasite-egg counts in most animals, oocyst spp. were 1.6x more likely (90% CI: 1.2043 – excretion was associated to age, being 2.1826, p=0.0014) to also be seropositive for animals younger than 12 months the most Anaplasma spp.. Dogs from regions outside infected (Mann-Whitney test p<0.001). The of the west were 0.0512x more likely (90% CI: results confirmed that parasite burden in 0.0118 – 0.2226, p=0.0001) to test seropositive France was comparable to other European for Ehrlichia spp.. Hunting dogs seropositive countries, regardless the heterogeneity of for B. burgdorferi were more likely to also farming practices and the gastrointestinal be seropositive for Anaplasma spp. (0.981 – parasite-control protocols among llama and 2.1225, p = 0.0569). alpaca herds. Conclusions: Region, due to the ticks that are found there, significantly influenced the PS01.69 Regional, Biological, and Exposure likelihood of hunting dog exposure to ticks Risk Factors That Drive Hunting Dog and tick-borne pathogens. Our results suggest Seropositivity to Tick-Borne Co-infections that hunting dogs were more frequently co- infected with tick-borne pathogens. Dogs had Kurayi Mahachi1, Eric Kontowicz1, Bryan significantly higher exposure to Babesia spp. Anderson1, Angela Toepp1, Adam Lima1, Mandy than previously reported, perhaps because it is Larson1, Geneva Wilson1, Tara Grinnage-Pulley1, not regularly diagnosed through rapid tests and Carolyne Bennett1, Marie Ozanne1, Michael is not susceptible to doxycycline treatment. Anderson1, Hailie Fowler1, Molly Parrish1, Jill Saucier2, Phyllis Tyrell2, Zachary Palmer1, Jesse Buch2, Ramaswamy Chandrashekar2, Breanna Scorza1, Grant Brown1, Jacob Oleson1, Christine Petersen1 1University Of Iowa, United States, 2IDEXX Laboratories Inc., United States

Background: Ticks and tick-borne diseases have increased both in range and rate of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 224 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS01.70 Oxifendazole Effect on Gene below detection levels for all tested genes Expression of P-glycoproteins in Two with the exception of L9 ribosomal protein, Haemonchus contortus Isolates Selected P-gps 2 and 3. Both resistant isolates for Resistance Against Ivermectin and presented similar low P-gp 3 expression in Oxifendazole comparison to untreated controls. P-gp 2 was also downregulated by oxifendazole Jomar Patricio Monteiro1,2, Janaelia Ferreira dosing but the ivermectin resistant isolate Vasconcelos2, Jessica Maria Leite dos Santos2, still maintained detectable levels of gene Gracielle Araújo Frota3, Marcel Teixeira1,2, expression. Cláudia Maria Leal Bevilaqua4, Luiz da Silva Vieira1 1Embrapa Caprinos e Ovinos, Brazil, 2Centro PS01.71 Haemonchus Contortus Infection Universitário INTA (UNINTA), Brazil, Modulates Lamb Ruminal Microbiome 3Universidade Estadual Vale do Acaraú (UVA), Composition and Function Brazil, 4Universidade Estadual do Ceará (UECE), Brazil Dr. Patricia Correa1,3, Dr. Lucas Mendes1, Ms. Leandro Lemos1, Dr. Emiliana Romagnoli1, Haemonchus contortus is the most prevalent Dr. Ana Sampaio1, Dr. Juliano Issakowicz1, small ruminant gastrintestinal nematode in Dr Concepta McManus2, Dr. Adibe Abdalla1, tropical areas. It is usually controlled using Dr. Siu Tsai1, Dr Antonio Faciola3, Dr. Helder anthelmintic drugs such as benzimidazoles Louvandini1 and macrocyclic lactones. Parasite resistance 1Centre for Nuclear Energy in Agriculture, against these compounds is currently University of São Paulo, Brazil, 2Institute of considered a global problem. P-glycoproteins Biological Sciences, University of Brasília, (P-gps) are ABC transporters involved with Brazil, 3Department of Animal Sciences, the removal of compounds from the cell University of Florida, United States interior and are known to be involved with drug resistance. In this study, we evaluated Haemonchus contortus (HC) is the most the gene expression of P-glycoproteins 2, important gastro enteric nematode in sheep 3 and 10 in H. contortus isolates selected in many regions of the world. However, the for ivermectin and oxifendazole resistance interaction between helminth and ruminal before and after oxifendazole dosing (5 microbiome remains largely unknown. Six mg/kg) in experimentally infected sheep. lambs fed exclusively Cynodon dactylon sp. Experimental animals (three per group) were hay were used in an experiment, divided in infected and, once infection was established, two treatments: artificially infected with 5,000 euthanized before and after dosing at the L3 (HC, n = 3) and control (n = 3) free of approximate peak of oxifendazole presence infection. Animals were followed for 34 days. in the blood stream. Adult male H. contortus Ovine ruminal contents were collected for were collected and total RNA extracted. DNA metagenomic sequencing to investigate Haemonchus contortus P-gps were identified the impact of HC infection on the structure based Caenorhabditis elegans orthologs and and dynamical function of the ruminal primers for reverse transcribed quantitative microbial community. Our results showed that PCR (RT-qPCR) were designed based HC infection increased Archaea and bacteria on sequences from genome assembly abundance in the rumen, but with no effect PRJEB506. RT-qPCR results were obtained on overall microbial diversity. The composition for the above mentioned P-gps and GAPDH, and functional analysis revelated less actin and ribosomal protein L9 as reference abundance (P < 0.05) of Thermomonospora, genes. There was no difference in expression Saccharomonospora, Leptospirillum genus, between both isolates for all tested genes as well as less abundance (P < 0.05) prior to exposure to oxifendazole with the of genes related to glycine, serine, and expception of P-gp 2 which was upregulated threonine metabolism and higher abundance in the ivermectin selected isolate. Gene (P < 0.05) of genes related to methane expression levels 8 hours after oxifendazole metabolism and stress response in the HC dosing led to a decrease in gene expression infected group. Our findings suggest that

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 225 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

the infection by HC may lead to a decrease more sensitive to ABZ and ugt-13 and ugt-12 of specific genera related to plant biomass make worms moderately sensitive to ABZ. degradation and may promote a differential By using CRISPR-Cas-9 knockout approach abundance of functional traits essential for we made a ugt-9 cluster (df) strain (~24 kb immunomodulation, microbiota restoration, deletion knocking out the tandem array of and the metabolism of essential amino acids seven ugt genes ) that showed a greater for protein synthesis in infected lambs. sensitivity to ABZ compared to each single loss-of-function mutant demonstrating that multiple ugt-9 cluster genes are involved in PS01.72 Ugt Genes Have Redundant Functions modulation of sensitivity to ABZ. in Modulating Benzimidazole Sensitivity in the Nematode Caenorhabditis Elegans PS01.73 Sub-Lethal Effects of Moxidectin Ms. Nidhi Sharma1, Ms. Vinci Au3, Dr. James on the Dung Beetle Onthophagus Landolti McGhee2, Dr. Susan Stasiuk2, Dr. Mark Edgley3, Harold (Coleoptera: Scarabaeinae) Dr. Don Moerman3, Dr. Paul Mains2, Prof. John Gilleard1 Prof. Roger Ivan Rodriguez-Vivas1, Gertrudis 1Faculty of Veterinary Medicine, University Basto-Estrella1, Enrique Reyes-Novelo2, of Calgary, Canada, 2Cummings School of William Arcila-Fuentes1, Melina M. Ojeda-Chi1, Medicine, University of Calgary, Canada, Imelda Trinidad-Martínez1, Imelda Martínez-M3 3Life Sciences Centre, University of British 1CCBA Universidad Autonoma De Yucatan, Columbia, Canada Mexico, 2Centro de Investigaciones Regionales “Dr. Hideyo Noguchi”, UADY, Mexico, The benzimidazoles (BZ) are a family 3Instituto de Ecología, A.C., Mexico of anthelmintics used in livestock and human parasitic nematode control. Drug Macrocyclic lactones can have adverse effects metabolism is an important modulator of in dung beetles exposed to manure containing drug efficacy in many systems including them. An evaluation was done of survival and insects and mammals. However, there has fecundity in Onthophagus landolti Harold been much less research in nematodes. adults fed manure from cows treated with Our previous work has shown that the free- moxidectin, and of imago emergence from living model nematode, Caenorhabditis the masses built by these adults. Three cows elegans and parasite Haemonchus contortus (Bos indicus x B. taurus) were subcutaneously detoxify BZ anthelminitics by conjugation injected with 1% moxidectin (0.2 mg kg-1 p.v.) with a glucose residue. This suggests the and another three were injected with 10% potential involvement of phase II xenobiotic moxidectin (1.0 mg/kg-1 p.v.). Manure was metabolizing enzymes (XMEs), specifically collected from these animals one day prior to the UGT enzyme family. The major objective moxidectin administration, at five days post- of this project is to identify and functionally treatment in the 1% and 10% treatments, and characterize nematode UGT enzymes that at fourteen days in the 10% treatment. Four metabolize ABZ in C. elegans. We undertook bioassays were done: a control using manure an RNAi screening of ugt genes and without moxidectin; 1% moxidectin at five identified that knockdown of ugt-9, ugt-11 days post-treatment; 10% moxidectin at five and ugt-22 make worms sensitive to ABZ days post-treatment; and 10% moxidectin treatment compared to wild type. Further we at fourteen days post-treatment. In each identified that RNAi of ugt-9 and ugt-11 may replicate, one pair of adult O. landolti was target other six ugt genes present in ugt-9 daily fed 30 g manure according to the cluster that are tightly liked and are closely treatments. No lethal effects were observed in phylogenetically related. We demonstrate the any of the four treatments. Sub-lethal effects redundant function of ugt-9 cluster genes in (P < 0.05) were present in the 10% moxidectin modulating ABZ potency in vivo by examining treatments at five and fourteen days post- the phenotype of seven loss-of-function ugt treatment. Fecundity was reduced by 78.2% mutants present in ugt-9 cluster. Specifically, at five days and 54.9% at fourteen days, and mutations in ugt-9 and ugt-11 make worms imago emergence was negatively affected at

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 226 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

both times. Current moxidectin application were compared by Tukey’s test (P <0.05). methods may have negative effects on the The counts of ticks for S. guianensis and S. cv environmental services provided by dung Campo Grande were 0.0 ± 0.0 and 20.0 ± 56.1 beetles, and therefore need to be modified and were significantly lower (P <0.05) when to minimize any impacts they might have compared to control B. ruziziensis (2587.0 ± on these vital members of tropical livestock 85.0). Percentage of larval mortality for S. systems. gracilis (53.3 ± 7.7) did not differ among other species (P> 0.05) but was greater (P <0.05) Research founded by CONACYT-Ciencia when compared to control B. decumbens (29.7 Básica (253578) grant. ± 10.2). More studies are needed in order to use Stylosanthes as part of sustainable control of R. microplus and H. contortus in pastures. PS01.74 Effect of Stylosanthes Species on Parasites of Cattle and Sheep PS01.75 Distribution Patterns of Sodium Luciana Katiki1, Bianca Tainá Azevedo1, Channel Mutations Associated With Guilherme Favero Luciani, Rodrigo Giglioti1, Pyrethroid Resistance and RDL Mutations Poliana Araújo Pacheco2, Bruna C Rodrigues Associated With Fipronil in Rhipicephalus Wedy1, Cristiane F Carvalho Fiorin1, Danielle Microplus Populations From Uruguay and Rio Rizatto Balbino1, Waldssimiler Teixeira de Grande Do Sul, Brazil Mattos1, Jessica Carolina Ribeiro1, Leandro Rodrigues1, Cecília José Veríssimo1 Eleonor Castro-Janer1, Dr. André Díaz1, Br. 1Institute of Animal Science (IZ/APTA), Brazil, Florencia Baraibar1, Florencia Fontes1, Dr. 2University of Sao Paulo, Brazil Magdalena Baraibar1, Dr. Tatiana Saporitti2, Guilherme M. Klafke3 Parasites cause losses in animal production. 1Udelar-School of Veterinary, Montevideo, The ectoparasite Riphicephalus microplus Uruguay, 2Instituto Nacional de Investigación and the endoparasite Haemonchus contortus Agropecuaria-Plataforma de Salud Animal, exposed to leguminous forages of the genus Colonia, Uruguay, 3Instituto de Pesquisas Stylosanthes which have phenolic compounds Veterinarias Desiderio Finamor, Secretaria da with known antiparasitic activity. In this Agricultura do Estado do Rio Grande do Sul, study, S. guianensis cv Mineirão, S. gracilis, Eldorado do Sul, Brazil S. bracteata, S. capitata, S. humilis and S. cv Campo Grande were evaluated in repellent Rhipicephalus microplus resistance to activity on R. microplus larvae and in mortality pyrethroids is widely dispersed and three on H. contortus larvae. Brachiaria ruziziensis nucleotide substitutions in the para- and B. decumbens were used as controls, sodium channel gene have been associated. respectively. The leguminous were harvested Resistance to fipronil is increasing in some from the agronomic field of Institute of countries and mutations in GABA-Cl gene Animal Science. For the repellency evaluation, were described in laboratory and field fipronil- triplicates of plants was exposed to 4,000 resistant strains. We developed a multiplex larvae of R. microplus. After 72 hours the allele specific-PCR (AS-PCR) to screen for tip of each triplicate were put inside Falcon® mutations in both genes of tick populations tubes and washed with 50 mL of distilled from Uruguay (n=31) and Rio Grande do Sul water. Ticks were count in five aliquots of 200 state, Southern Brazil (n=14). Toxicological L. For anthelmintic evaluation,1 g of each in vitro bioassays with larvae and adults plant was placed in Petri dishes in triplicates were used to determine susceptibility to withμ 15 mL distilled water with 1,000 L3 H. cypermethrin, flumethrin and fipronil. The contortus and incubated 24 hours/27°C. The AS-PCR included the detection of the three plants were washed with distilled water to a nucleotide substitutions the para-sodium final volume of 50 mL and larvae of 10% of channel gene (C190A; G215T and T2134A) this volume were counted as dead or alive. and a non-synonymous mutation in the Statistics were performed with PROC GLM GABA-Cl gene (G858T/C859C). The mutation (SAS), and the means of the different plants C190A was present in all pyrethroid-resistant

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 227 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

populations from Uruguay and Brazil, most which had not been treated in the previous frequently homozygous (30.9%). The mutations 2 months. Faecal Egg Count was performed G215T and T2134A were not detected. Of 12 with Mini-FLOTAC technique to select the fipronil-resistant populations, 10 presented 2-3 faeces with the highest FEC values and at least one mutant allele, more frequently in to make a bulk faecal sample. In addition heterozygosis. Nevertheless, in two fipronil- to coproculture, Egg Hatch Assay was susceptible populations, the mutation performed as follows: i) eggs were isolated G858T/C859C was detected. The present by sieving, centrifugation and flotation in survey shows a high frequency of pyrethroid saturated NaCl solution and collected with a resistance mutations widely dispersed in the sucrose step gradient centrifugation, ii) eggs region. The absence of the G215T and T2134A were then tested using 7 concentrations of could be suggest a geographic separation thiabendazole (TBZ) from 0.01 to 0.5 g/ of C190A. Other mechanisms besides target mL to determine EC50 (cut-off: 0.1 g/mL). site insensitivity may be involved in fipronil In the 17 farms, EC50 values ranged fromµ resistance, since in some populations the 0.027 to 0.051 g TBZ/mL and indicatedµ mutation was not detected. the susceptibility of the worm populations. Coprocultures resultsµ showed a predominance Acknowledgements: ANII-FSA 1-2013-13028, of Cooperia infective larvae. The allele INIA-PSA, CNPq, FAPERGS. frequency of BZ-resistance associated beta-tubulin isotype 1 single-nucleotide- polymorphisms within pooled samples of each PS01.76 Benzimidazole Resistance Survey of the 17 examined will be documented using in Gastrointestinal Nematodes of Dairy and a previously established pyrosequencing- Beef Cattle in Western France Using Egg based assay. The results of this study will Hatch Assay have to be confirmed on a larger scale but suggest, in contrast to small ruminants, that Dr Christophe Chartier1, Dr Nadine Ravinet1,, BZ anthelmintics are fully effective in cattle. Marie Bertocchi1,, Léa Hirschauer1,, Dr Helga Waap2, Dr Georg van Samson-Himmelstjerna3 1BIOEPAR, INRA, ONIRIS, Nantes, France, PS01.77 Selective Sweep and Phylogenetic 2National Institute for Agrarian and Veterinary Models for the Emergence and Spread of Research, Oeiras, Portugal, 3Institute for Pyrimethamine Resistance Mutations in Parasitology and Tropical Veterinary Medicine, Plasmodium Vivax Freie Universität, Berlin, Germany Umer Chaudhry1, Ayaz Shaukat3, Qasim Ali2, There is a general consensus on the Timothy Connelley1, Muhammad Azmat Ullah widespread anthelmintic resistance (AR) Khan3, Mushtaq A. Saleem3, Mike Evans1, Imran of gastrointestinal nematodes in Europe in Rashid2, Neil D. Sargison1 small ruminants. There is much less data 1University Of Edinburgh, United Kingdom, available regarding AR in cattle, although 2University of Veterinary and Animal Sciences reports of lack of efficacy of macrocyclic Lahore, Pakistan, 3University of Central lactones in several European countries Punjab, Lahore, Pakistan have been published recently. Concerning benzimidazoles (BZ), only one survey on 12 Pyrimethamine resistance is a major concern herds in Germany and Sweden is available and for the control of human haemoprotozoa, indicated a full effectiveness of albendazole. especially Plasmodium species. Currently, Our study aimed at assessing the anthelmintic there is little understanding of how efficacy of BZ in dairy/beef cattle in western pyrimethamine resistance developed France using the Egg Hatch Assay (EHA). in Plasmodium vivax in the natural field During summer 2018, 17 cattle farms were conditions. Here, we present for the first- selected as a convenience sample. Individual time evidence of positive selection pressure faecal samples were collected and stored on a dihydrofolate reductase locus and its anaerobically in approximately 10 first or consequences on the emergence and the second grazing season animals per farm spread of pyrimethamine resistance in P. vivax

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 228 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

in the Punjab province of Pakistan. First, we levels of parasite resistance in nematodes of examined the dihydrofolate reductase locus sheep. Changes in drug target sites and the in 38 P. vivax isolates to look for evidence of up-regulation of detoxification systems seem positive selection pressure in human patients. to be implicated in this phenomenon. Several The S58R (AGA)/S117N (AAC) double mutation studies have shown that ATP-binding-cassette was most common, being detected in 10/38 (ABC) transporters such as P-glycoprotein isolates. Single mutation S117N (AAC), I173L (P-gp) play an important role in multidrug (CTT) and S58R (AGA) SNPs were detected resistance in many organisms, including in 8/38, 2/38 and 1/38 isolates, respectively. several nematode species. The goals of the The F57L/I (TTA/ATA) and T61M (ATG) SNPs current work were: 1) to compare the gene were not detected in any isolates examined. expression of several cellular transporters Although both soft and hard selective sweeps in both a susceptible (S-IVM) and a highly have occurred with striking differences between ivermectin (IVM)-resistant Haemonchus isolates, there was a predominance of hard contortus isolate (HR-IVM); 2) to assess the sweeps. A single resistance haplotype was effect of IVM on ABC transporters expression present at high frequency in 9/14 isolates, patterns in the HR-IVM isolate under in vivo providing a strong evidence for single conditions. To this end, the transcriptional emergence of resistance by the single mutation, levels of ABC transporters in adult HR-IVM H. characteristics of hard selective sweeps. contortus recovered from IVM-treated lambs In contrast, 5/14 isolates carried multiple (2 mg/kg) at 12 and 24 hours post-treatment, resistance haplotypes at high frequencies, were compared to those obtained from the providing an evidence of the emergence S-IVM and HR-IVM specimens collected from of resistance by recurrent mutations, untreated lambs. The phylogenetic tree with characteristics of soft selective sweeps. Our the transporter sequences of the reference phylogenetic relationship analysis suggests that nematode Caenorabditis elegans and H. S58R (AGA)/S117N (AAC) and S117N (AAC) contortus allowed us to found the putative mutations arose multiple times from a single orthologous genes P-gp 1, 2, 3, 9.1, 10, 11, 13, origin and spread to multiple different cities 16 and 17; MRP 3, 4, 7 and 8; and Haf 2, 3, 4, in the Punjab province through gene flow. 6 and 9. Next generation sequencing analysis Interestingly, the I173L (CTT) mutation was showed that both H. contortus isolates present on a single haplotype, suggesting that it express 6 of the 9 P-gps, 3 of the 5 multi- arises rarely and has not spread between cities. drug resistant proteins (MRPs) and 4 of the Our work shows the need for responsible use of 5 Half (HAF) transporter genes. Some of existing and new antimicrobial drugs and their these ABC transporter genes are differentially combinations, control the movement of infected expressed in the S and HR isolates. IVM patients and mosquito control strategies. treatment induced slight changes in the mRNA levels of MRP-4 and P-gp transporters, but the biological significance of the observed PS01.78 Transcriptomic Analysis of ABC- changes may not be enough to explain the Transporters (P-GP, MRP and HAF) in high level of IVM resistance displayed by the Haemonchus Contortus Isolates With isolate under study in the current trial. Different Susceptibility to IVM

Laura Maté1, Mariana Ballent1, Candela Cantón1, PS01.79 Sheep, Strongyles and Sequencing: Laura Ceballos1, Prof. Adrián Lifschitz1, Prof. Investigating Ivermectin Resistance in UK Carlos Lanusse1, Paula Domínguez1, Prof. Luis Field Populations Alvarez1, Pedro Lirón1 1Laboratorio de Farmacología, Centro de Miss Jennifer McIntyre1, Dr. Roz Laing1, Ms. Investigación Veterinaria Tandil (CIVETAN) Kirsty Maitland1, Mr. James McGoldrick1, Dr. UNCPBA-CICPBA-CONICET, Campus Stephen Doyle3, Dr. James Cotton3, Ms. Nancy Universitario, Tandil, Argentina Holroyd3, Prof. Eric Morgan4, Dr. Hannah Vineer5, Miss Katie Bull6, Dr. David Bartley2, Inadequate and intensive use of anthelmintic Mrs. Alison Morrison2, Miss Kim Hamer1, Prof. compounds has led to the emergence of high Neil Sargison7, Prof. Eileen Devaney1

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 229 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

1University of Glasgow, United Kingdom, PS01.80 Treatment of Canine and Feline 2Moredun Research Institute, Edinburgh, Gastrointestinal and Respiratory Helminths United Kingdom, 3Wellcome Sanger Institute, Using a Combination of Milbemycin Oxime Hinxton, United Kingdom, 4Queen’s University and Praziquantel (Milpro® Tablets, Virbac) in Belfast, Belfast, United Kingdom, 5University Greece of Liverpool, Liverpool, United Kingdom, 6University of Bristol, Bristol, United Kingdom, Dr. Athanasios Angelou1, Dr. Konstantinos 7Royal (Dick) School of Veterinary Studies, Arsenopoulos1, Dr. Isaia Symeonidou1, Edinburgh, United Kingdom Dr. Athanasios Gelasakis1, Dr. Emmanuel Briant2, Dr. Stephane Bonneau3, Dr. Elias Parasitic gastroenteritis is a major production Papadopoulos1 limiting disease of small ruminants 1Aristotle University of Thessaloniki, worldwide, and anthelmintic resistance is Thessaloniki, Greece, 2Virbac Global Medical steadily increasing. The abomasal nematode Department, Carros, France, 3Virbac R and D Teladorsagia circumcincta is the primary Department, Carros, France pathogen on most sheep farms in the UK during the summer months, when lambs are Dogs and cats are the most common pets in their primary growth phase. Ivermectin living in close relationship with humans, is commonly used to treat infected sheep offering several benefits. Both animal throughout the year, but resistance is highly species can harbour numerous helminths, prevalent; recent studies in the UK and Ireland which may affect their health and wellbeing, have demonstrated rising numbers of farms while some of them may have zoonotic with detectable ivermectin resistance by implications. The aim of this study was to faecal egg count reduction test, a concern for evaluate the effectiveness of the combination sustainable control of parasites in the future. of milbemycin oxime and praziquantel There are many limitations to the faecal egg (Milpro, Virbac) against gastrointestinal and count reduction test, and such limitations, respiratory helminths in dogs and cats in combined with a highly complicated disease, Greece. Towards this end, faecal samples make understanding the effect of different were collected for coprological examination management practices on the development from 728 dogs and 670 cats from different of anthelmintic resistance difficult. Ivermectin parts of the country, targeting potentially resistance in T. circumcincta is poorly infected animals. A questionnaire was understood, and molecular tests are needed. accompanying the sample with relevant In this study we have looked at two UK farm information (sex, age, lifestyle, breed, owner populations of T. circumcincta pre- and post- education etc). Overall, 63.2% (460/728) and ivermectin treatment using next generation 41.8% (280/670) sampled dogs and cats, sequencing techniques (ddRAD-Seq and respectively, were detected to excrete at least Pool-Seq) to identify regions of the genome one parasitic element. Infected animals were under selection. Despite the fragmented treated at D0 with the above combination and nature of the T. circumcincta draft genome a second sample was tested again at D30 to assembly, we found a single large locus to evaluate the effective removal of the parasitic be under ivermectin selection, in addition elements (% efficacy). Dogs were found to to many smaller loci. Though some selected be infected with Toxocara canis (24.9%, mean loci appear farm specific, several of the same efficacy 98.7%), hookworms (19.6%, efficacy contigs are under selection on both farms. 100%), Trichuris vulpis (12.1%, efficacy 93.7%), Previous candidate ivermectin resistance Toxascaris leonina (8.7%, efficacy 100%), genes from the literature do not appear to be Dipylidium caninum (4.9%, efficacy 99.7%) under selection so are unlikely to contribute and Taenia spp. (0.5%, efficacy 97.7%). Cats to ivermectin resistance in these populations. were found to be infected with Toxocara cati (31%, efficacy 99.5%), hookworms (21.3%, efficacy 100%), lungworms (5.7%, efficacy 92.4%) and Dipylidium caninum (1.6%, efficacy 99.3%). In all cases of reduced efficacy, a second treatment was carried out at D30 and

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 230 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

retested at D60 leading to a 100% parasite with similar kinetics as a typical experimental removal. The most important risk factor was mite infestation. Topical application of found to be lifestyle and owner education. carvacrol on shaved skin caused mild and It was concluded that the combination of transient erythema 20 min after treatment. No milbemycin oxime and praziquantel (Milpro, other side effects were observed. Considering Virbac) offers an effective solution to control the strong acaricidal activity of carvacrol in common gastrointestinal and respiratory vitro and in vivo and the mild and transient helminths in dogs and cats in Greece. local side effects after topical treatment, carvacrol shows potential as an acaricidal agent in the treatment of P. ovis in cattle. PS01.81 Acaricidal Activity of Plant-Derived Essential Oil Components Against Psoroptes Ovis In Vitro and In Vivo PS01.82 Evaluation of Levamisole Pharmacokinetics and Milk Excretion in Dairy Dr. Zhenzhen Chen1, Dr. Wouter van Mol1, Dr. Goats Marieke Vanhecke1, Prof. Luc Duchateau2, Prof. Edwin Claerebout1 Miss Jorgelina Lúpori1, Dr. Juan Sallovitz1, Mrs. 1Laboratory Of Parasitology, Faculty of Paula Dominguez1, Mrs. Cristina Farias1, Mrs. Veterinary Medicine, Ghent University, Ghent, Gabriela Cadenazzi1, Dr. Carlos Lanusse1, Dr. Belgium, 2Department of Nutrition, Genetics Fernanda Imperiale1 and Ethology, Faculty of Veterinary Medicine, 1Laboratorio de Farmacología, Centro de Ghent University, Ghent, Belgium Investigación Veterinaria de Tandil (CIVETAN), UNCPBA-CICPBA-CONICET. Facultad de Psoroptes ovis is a major health problem Ciencias Veterinarias, UNCPBA., Tandil, in beef cattle. Treatment is limited to local Argentina administration of amitraz or pyrethroids or systemic administration of macrocyclic Introduction: Levamisole (LVM) is a lactones. Treatment failures with macrocyclic nematodicidal compound available for lactones have been reported in recent veterinary medical use for over 50 years. years. To investigate potential alternative Since the widespread development of treatments, the acaricidal activity of four drug resistance is a serious productive plant-derived essential oil components, i.e. inconvenient, LVM is currently used extralabel geraniol, eugenol, 1,8-cineol and carvacrol as a therapeutic alternative in dairy sheep against P. ovis was assessed in vitro and and goats. Thus, information on drug residual in vivo. Three components showed a concentrations in milk is needed to assure concentration-dependent acaricidal activity protect consumer`s safety. The current work in a contact assay, with LC50 of 0.56 %, 0.38 aimed to evaluate the concentration profiles % and 0.26 % at 24 h for geraniol, eugenol, of LVM in plasma and its pattern of milk and carvacrol, respectively. In a fumigation excretion in lactating dairy goats treated at bioassay, carvacrol demonstrated the best different stages of lactation. efficacy as it killed all mites within 50 min after treatment, whereas geraniol, eugenol, Material and Methods: Twelve (12) female and 1,8-cineol needed 90 min, 150 min, Saanen dairy goats at early-mid stage of and 90 min, respectively. Following a 72 lactation (group A) and at mid-late stage of h incubation period in a residual bioassay, lactation (group B) were orally treated with eugenol and carvacrol killed all mites after 4 h LVM (7.5 mg/kg) (Ripercol®, Zoetis). Blood of exposure to LC50 and LC90, while geraniol and milk samples were collected between 0 killed all mites only after 8 h exposure at and 5 days post-treatment to characterize LC50. Topical treatment with 2 % carvacrol in the plasma and milk disposition kinetics. Tween-80 of six calves with experimental P. LVM concentrations in plasma and milk were ovis infestations, reduced mean mite counts determined by HPLC with UV detection. by 98.48±2.36 % at 6 weeks post treatment. Results: LVM parent compound was detected In the control group which was treated with in plasma and milk up to 8 h and 12 h post- Tween-80 only, the mite population increased treatment, respectively. Plasma concentrations

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 231 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

increased progressively to a maximum from 2154 ng/g to 225 ng/g; mean IVM concentration (Cmax) of 0.89 ± 0.2 g/ml at concentration in ryegrass ranged between 0.27 h (group A) and 0.77 ± 0.3 g/ml at 0.2 378.65ng/g and 21,74ng/g. Strikingly, clover h (group B). LVM milk residual concentrationsµ development was delayed until 30 dpt and were lower than measured in plasma.µ IVM concentration in this specie ranged between 94,09 ng/g and 4,56ng/g. Significant Conclusion: The pharmacokinetic results differences were detected between species reported here confirm that LVM is excreted (p=0,0374). In the LG, IVM concentration by milk in lactating dairy goats. The was between 22,26ng/g and 1,02ng/g in residual concentrations in milk (0.05 g/ml) ryegrass, and between 10ng/g and 1,02ng/g detected up to 12 h, should be considered in clover, without statistically significant before issuing any recommendation onµ the differences between species (p=0,8301). Trial manufacturing of milk from dairy goats under 2: IVM was detected in both plant species at antiparasitic treatment with LVM. significant levels (p>0,05) in all the sampling times; mean IVM concentration was between 10ng/g and 5ng/g in radish, and 17,70ng/g PS01.83 Uptake of Ivermectin from Growing and 6,55ng/g in lettuce. IVM concentration Substrate to Plant Species in the substrate decreased from 1311ng/g to 116ng/g. In conclusion, IVM concentrations in Lucía Iglesias1, Juan Sallovitz2, Carlos Saumell1, soil or composted substrate are transferred Silvina Fernandez1, Milagros Junco1, Federica to plants during growth period, and could Sagues1, Zegbi1, Adrián Lifschitz2 be incorporated into the food chain of both 1Área de Parasitología y Enfermedades livestock and humans. Parasitarias, Centro de Investigación Veterinaria de Tandil (CIVETAN) (UNCPBA- CICPBA-CONICET), Facultad de Ciencias PS01.84 The Efficacy of a Proprietary Veterinarias, UNCPBA, Tandil, Argentina, Formulation of Imidacloprid 10% + 2Laboratorio de Farmacología, Centro de Moxidectin 1% (Advantage Multi®, Advocate®) Investigación Veterinaria de Tandil (CIVETAN) Spot-On for the Treatment Against (UNCPBA-CICPBA-CONICET), Facultad Microfilariae of Brugia Pahangi in Naturally de Ciencias Veterinarias, UNCPBA, Tandil, Infected Cats Argentina Dr. Tawin Inpankaew1, Miss Pornkamol Ivermectin (IVM) is a worldwide-used Phoosangwalthong1, Dr. Susanne Siebert2, Dr. antiparasitic drug. However, its high level of Roland Schaper2 faecal elimination together with its transfer 1Faculty of Veterinary Medicine, Kasetsart from dung pats to the underlying soil as well University, Bangkok, Thailand, 2Bayer Animal as the common practice of using manure Health GmbH, Leverkusen, Germany for soil amendement represents a potencial risk to plants growing in these substrates. Lymphatic filariasis (LF) is one of the most Two trials were conducted to evaluate the debilitating neglected tropical diseases uptake of IVM to: 1) a crop of ryegrass (Lolium (NTDs). It is caused by parasitic worms multiflorum) and clover (Trifolium repens) transmitted to humans by mosquitoes. Recent growing for 120 days post treatment (dpt) evidence has also indicated that Brugia in IVM-spiked soil at 3000 (High group, HG) pahangi, a filarial nematode that is naturally and 90ng/g (Low group, LG); and 2) a crop found in cats, can cause clinical infection of radish (Raphanus sativus) and lettuce in humans, with clinical presentations that (Lactuca sativa) growing for 60 dpt in a are consistent with lymphatic filariasis. mix of soil and 10% IVM-spiked manure at Imidacloprid 10% + Moxidectin 1% has more 3000ng/g. Soil, soil-manure mix and plants recently been introduced as a topical spot- were sampled starting at 15 dpt and at the on, and marketed worldwide as Advantage end of each trial. All matrices were analyzed Multi® or Advocate® by Bayer Animal Health by HPLC to quantify IVM concentration. Trial (Leverkusen, Germany). However, the efficacy 1: In HG, IVM concentration in soil decreased against infections with B. pahangi in cats

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 232 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

has never been tested. The goal of this 0.39mg/ml (30.89%) reduction in the egg project was to determine the efficacy of the count as compared to the reference dose combination of imidacloprid 10% + moxidectin (Oxfendazole) concentration 0.01415mg/ml 1%against microfilariae of B. pahangi in (78% ) , 0.00707mg/ml (68%) , 0.003537 naturally infected cats. Twelve cats naturally mg/ml (68.4%) ,0.001768 mg/ml (47.06%) infected with B. pahangi were divided into ,0.00088 mg/ml (30.89%) , 0.00043mg/ treatment and control groups. Cats in the ml (27.81) and 0.000215 mg/ml (30.15%) treatment group were given a combination reduction in the eggs. So the herbal complex of imidacloprid (10 mg/kg bodyweight) and showed a graded response as anthelmintic moxidectin (1 mg/kg bodyweight) spot on so Herbal complex seems to be promising. monthly for 5 months. Microfilariae were The trials on the large scale for the efficacy counted weekly for a period of 20 weeks for and safety, however, are recommended both groups. The mean microfilaria counts before the herbal complex is considered for decreased significantly in the treatment commercialization. group after 3 month application. Moreover, all treatment cats were negative for microfilaria count on week 15, 18 and onward PS01.86 Morphological and Molecular after 4 month application. In this study, Characterization of Feline Filarid Nematode cats naturally infected with B. phangi were and Its Zoonotic Implication successfully treated using 5 monthly doses of a imidacloprid 10% + moxidectin 1% spot on Prof. Ananda Konanur Javaregowda1, Prof. formulation. Malatesh D. S.1, Manjunath Reddy G.B.3, Lavanya K.V.4, Scolar Sumana K3 1Department of Veterinary Parasitology, PS01.85 Anthelmintic Activity of A Herbal Karnataka Veterinary, Animal And Fisheries Complex Against Heamonchus Contortus Sciences University (b), Veterinary C, Shimoga, India, 2Department of Veterinary Ms. Warda Qamar1, Muhammad Arfan Zaman1, Medicine, Karnataka Veterinary, Animal And Mr. Zohaib Shahid1, Ms. Uzma Mehreen1, Mr. Fisheries Sciences University (b), Veterinary Kazim Ali1, Fiaz Qamar1 C, Shimoga, India, 3NIVEDI, ICAR Research 1CVAS, Jhang, Pakistan Institute , Yelahanka, Bangalore, India, 4Department of Veterinary Parasitology, The study was carried out to evaluate Karnataka Veterinary, Animal And Fisheries anthelmintic properties of a herbal complex Sciences University (b), Veterinary C, Hebbal, (Oregano essential oil, Cinnamon essential Bangalore, India oil , Rosemary essential oil and Red chilli powder) against Heamonchus contortus a Dirofilariosis is a potential zoonotic disease common pathogenic nematode effecting caused by filarid nematodes, prevalent in the ruminant by sucking blood from the several parts of the world transmitted mainly abomasum resulting to anemia and dullness by mosquito vectors. In the present study, which leads to low productivity and mortality. the filarid worms were recovered from the The herbal complex with concentration subcutaneous tissues of seven cats during (25mg/ml to 0.39mg/ml) was exposed to the sterilization in mangalore, a coastal region heamonchus eggs in the lab with 136egg/ of Karnataka. The worms were collected in 0.3ml concentration following the standard PBS and identified as D. repens based on the procedure of egg hatch test. The reference morphometry. Morphologically, adult worms drug used in the study was oxfendazole showed longitudinal ridges on the cuticle with with the concentration of 0.01415mg/ml anterior muscular and posterior glandular to 0.000215mg/ml. After incubation of 48 type of oesophagus. The length and width of hours the results recorded for the egg count female worms were in the range of 112 to 130 in which the herbal formulation used with mm and 4.1 to 5.8 mm in diameter whereas, 25mg/ml ( 96.70%), 12.5mg/ml (91.13%) , male worms of 44 to 58 mm length and a 6.25mg/ml (71.30%),3.125mg/ml (44.12%), diameter of 3.2 to 4.1 mm respectively. The 1.56mg/ml (41.18%) ,0.78mg/ml (33.83%) and blood samples of cats showed microfilaria

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 233 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

by modified knott’s method (MKM) and of Cytauxzoon felis. Sixteen cats were dosed quantitative buffy coat (QBC). Morphologically, with selamectin/sarolaner or a placebo the microfilariae were unsheathed with blunt (vehicle control) on Days 0, 28, and 56. In head and the tail was long, curved with hook phase 1, each cat was infested with 50 (±5) like posterior end. The length and width of unfed adult A. americanum on Day 4 and microfilaria were in the range of 310.9±9.10 m tick counts were conducted on Day 6 (48 and 6.51±0.14 m respectively. Based on hours post-infestation) and Day 7 (72 hours morphometry, the microfilariae were belongsµ post-infestation). In phase 2, each cat was to D. repens. Duringµ the study, a human infested on Day 60 with 50 (±5) adult A. filarid worm from the said region received for americanum acquisition-fed as nymphs on identification and the morphometry showed two C. felis-infected donor cats. Tick counts similar to that of D. repens. For confirmation, were conducted on Day 62 (48 hours post- DNA from both cat and human filarid worms infestation) and Day 63 (72 hours post- including microfilariae were extracted and infestation). Placebo cats were adequately subjected to PCR using pan-filarial primer pair infested on all count days, with least squares of DIDR-F1 and DIDR-R1 targeting ITS2 of the (geometric) mean live tick counts ranging ribosomal DNA of D. repens. The PCR yielded from 34.0 (28.8) to 46.1 (46.0). Treatment amplicons were sequenced and analysed. The reduced the least squares (geometric) adult worms of cats, human and microfilariae mean counts compared to placebo by 27.1 of cats from coastal region of Karnataka (32.1)% and 90.4 (96.8)% on Days 6 and 7, were confirmed as D. repens based on PCR respectively. The corresponding percent targeting ITS2 genes. The nucleotide sequence reductions were 56.4 (60.6)% and 94.7 (97.3) analysis obtained showed more similarity with % on Days 62 and 63, respectively. Least D. repens sequences than D. immitis. squares mean counts were significantly lower in the treated group compared with the placebo group on all count days (P≤0.0286). Poster Session 02 In phase 2, seven cats in the control group and no cats in the selamectin/sarolaner) group July 9, 2019, 10:00 – 16:30 became infected with C. felis (P=0.0017). Exhibit Hall A, July 9, 2019, Level 1 Topical treatment with selamectin/sarolaner (Revolution® Plus) was >90% effective in reducing A. americanum tick counts 72 PS02.01 Efficacy of a Topical Formulation of hours after infestation and prevented the Selamectin Plus Sarolaner (Revolution® Plus) transmission of C. felis from infected ticks Against Induced Infestations of Amblyomma following administration of three consecutive Americanum on Cats and Prevention of monthly treatments. Extracted from https:// Cytauxzoon Felis Transmission doi.org/10.1016/j.vetpar.2018.10.018. Mason Reichard1, Jady Rugg2, Jennifer Thomas1, Kelly Allen1, Annie Barrett1, Jill Murray1, Brian Herrin1, Rachel Beam1, Vickie King2, Adriano Vatta2 1Oklahoma State University, Stillwater, United States, 2Zoetis, Kalamazoo, United States

Our aims were to determine the efficacy of a selamectin (6.0 mg/kg) plus sarolaner (1.0 mg/kg) combination (Revolution® Plus / Stronghold ® Plus, Zoetis) applied topically once a month on cats for three consecutive months against induced infestations of Amblyomma americanum adults and to evaluate the effectiveness of the product in preventing the transmission

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 234 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.02 Roles of Flies and Beetles INTRODUCTION: LongRange® (eprinomectin Associated with Animal Agriculture in extended-release injection) provides Salmonella Maintenance and Transmission therapeutic activity against developing, inhibited and adult nematodes as well as Prof. Nancy C. Hinkle1, Aubree Kelly2, Yumin prevention from infection with a range of Xu3, Dr. Sha Tao4, Dr. Mark Harrison5, Dr. Jinru gastrointestinal nematodes and lungworms Chen3 for up to 150 days. However, data are 1Dept. of Entomology, University of Georgia, lacking on its efficacy against developing Athens, United States, 2Centers for Disease and adult Bunostomum phlebotomum, Control, Atlanta , United States, 3Dept. of Haemonchus contortus and Nematodirus Food Science & Technology, Univ. of Georgia, helvetianus, and developing Haemonchus Griffin, United States,4 Dept. of Animal and placei, Oesophagostomum radiatum and Dairy Science, Univ. of Georgia, Tifton, United Trichostrongylus colubriformis. For this States, 5Dept. of Food Science & Technology, reason, nine blinded studies compliant to Univ. of Georgia, Athens, United States GCP and VICH were conducted in the US and Germany to assess the efficacy for each Salmonellosis affects over 1.2 million species and stage. Americans annually, with 65 of every 100,000 children under the age of 5 years suffering METHODOLOGY: 218 young cattle, 180 with from the illness. There are many means by induced and 38 with naturally acquired which Salmonella infection is acquired, but nematode infections were included in seven research is demonstrating that arthropods and two studies, respectively. In each study, are frequent Salmonella carriers. Flies, not cattle were formed randomly into groups surprisingly, are commonly contaminated of nine or ten animals each, which served with Salmonella, and their close association either as saline-treated controls or received with humans and human food makes them LongRange® (1 mL per 50 kg bodyweight) likely means of transmission. Their mobility when the parasites were fourth-stage enhances their vectorial significance. A developing (L4) or adult nematodes. Efficacy common pest of poultry production, the lesser was determined based on geometric means mealworm (also known as the poultry darkling of nematode counts established following beetle) not only acquires and transmits necropsy of animals. Salmonella as both larvae and adults, but is capable of maintaining Salmonella infection RESULTS: Treatment was well accepted and through pupation, emerging as Salmonella- no treatment-related problems were observed infected adult beetles. Chickens can acquire in any study. Counts of each species and Salmonella by eating infected beetle larvae stage of nematodes were significantly lower or adults. Thus, management of Salmonella in the LongRange®-treated cattle than in the exposure necessitates suppression of these controls. Percent efficacy for each study by arthropods involved in maintenance and species was 100% against L4 and adult B. transmission of the pathogen. phlebotomum; 69.6% and >99% against L4 and ≥90.8%, >99% and 100% against adult H. contortus; 91.4% and 97.4% against L4 H. PS02.03 Use of LongRange® (Boehringer placei, >99% and 100% against L4 and >99%, Ingelheim Animal Health) Placei and >99% and 100% against adult N. helvetianus; Nematodirus Helvetianus, Oesophagostomum >99% and >99% against L4 O. radiatum; and Radiatum and Trichostrongylus Colubriformis 98.9% and 100% against L4 T. colubriformis. Infections in Cattle Results of these studies demonstrated that Martin Liebstein1, Steffen Rehbein1, Bruce LongRange ® is an efficacious treatment Kunkle1, Jeffrey Shryock1, Jonatan Bader1, against a broad spectrum of developing Dietmar Hamel1, Martin Visser1, Stephen Yoon1, larval and adult nematode endoparasites. The Becky Fankhauser1 efficacy demonstrated was similar to what 1Boehringer Ingelheim Animal Health, Fulton, was previously reported against the most United States widespread nematodes.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 235 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.04 Targeted Selective Treatment This study was supported by Swedish Against Liver Flukes (Fasciola Hepatica) in Research Council (FORMAS) Nr. 2016- Dairy Herds in Sweden 00510 and OP VVV Project CZ.02.1.01/0.0/ 0.0/15_003/0000495 FIT (Pharmacology, Giulio Grandi1, Bengt-Ove Rustas1, Niclas Immunotherapy, nanoToxicology). Högberg1, Adam Novobilsky2, Johan Höglund1 1Swedish University of Agricultural Sciences, Uppsala, Sweden, 2Veterinary Research PS02.05 Trypanosomiasis in Sedentary Institute, Brno, Czech Republic Cattle at Previously Assumed Trypanosoma- Free Jos Plateau, Nigeria Control of fasciolosis is primarily based on anthelmintic treatment in combination with Prof. Maxwell Opara1 grazing routines. In dairy cows, flukicide 1University Of Abuja, Abuja, Nigeria treatment is restricted due to concerns about drug residues in milk. Although the impact of Trypanosomiasis in sedentary Cattle at fluke infection on dairy production is under previously assumed Trypanosoma-free Jos debate, the costs of fasciolosis and benefits plateau, Nigeria of deworming have never been evaluated in Obeta, S. S., Mohammed, B. R., Simon, K. M., Sweden. Jegede, O. C. and Opara, M. N*. Protozoal and Zoonotic Parasites Research The aim of this study was to evaluate the Group efficacy of a targeted anthelmintic treatment Department of Veterinary Parasitology and by investigation of the epidemiology of Entomology liver fluke infection and its impact on dairy University of Abuja, Nigeria production. *Corresponding Author: opara.maxwell@ gmail.com Four fluke-infected dairy herds in southwest Sweden were monitored for Fasciola hepatica Abstract antibodies in milk and/or faecal coproantigens The occurrence of trypanosomiasis was between 2017 and 2018. In addition, data on investigated in sedentary Bunaji (Bos indicus) milk yield and quality were collected on a cattle grazing at 5 different villages of Barkin monthly basis between 2016 and 2018. The Ladi Local Government Area of Plateau State, deworming protocol was oral drenching of all Nigeria. Two hundred cattle were examined, non-lactating animals (heifers and dry cows) 76 (38.0%) of which had trypanosome with albendazole (10 mg/kg bodyweight) on infection. Three species, Trypanosoma vivax three different visits both during winters 2017 (78.9%), T. Congolense (15.8%), and T. brucei and 2018. On each visit, dewormed animals (5.3%) were encountered. More males (42.9%) along with 15 individual milking cows were also than females (31.8%) were affected while sampled for the detection of coproantigens. animals of 3-5 years age cohort had the Furthermore, milk samples were collected highest infection rate of 46.7%. The existence quarterly from the whole herd and examined of trypanosome infections in sedentary cattle for antibody levels using an indirect ELISA. populations of the Jos Plateau seems to Total prevalence in dairy herds based on suggest that the area which was previously coproantigen varied from 27 to 85%. However, assumed to be Trypanosoma-free may have in all four herds, heifers had lower F. hepatica acquired infection status. Suggestions on the prevalence than milking cows. Deworming cause of this shift are presented in study. in 2017 resulted in a significant decrease of coproantigen-positivity but only in the two herds with higher fluke burden in 2018. Nevertheless preliminary data show a significant increase of energy corrected milk only on one of these farms. More detailed statistical modelling is needed before final conclusion about the impact of liver flukes on dairy cattle.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 236 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.06 Profitability of Parasiticides for PS02.07 Risks with Next Generation Control of Horn Flies (Diptera: Muscidae) in Sequencing in the Detection of Strongylus Stocker Cattle Vulgaris

Dr. Justin Talley1, Dr. Wade Brorsen1, Dr. Dale Alice Anlind1, Peter Halvarsson2, Johan Blasi2, Dr. Brian Adam2 Höglund2 1Oklahoma State University, Stillwater, United 1Vidilab Ab, Enköping, Sweden, 2Swedish States, 2Kansas State University, Manhattan, University of Agricultural Sciences, United States Department of Biomedical Science and Veterinary Public Health, Uppsala, Sweden Horn flies (Haematobia irritans) have been estimated to cause over US $2 billion per Strongylus vulgaris is considered as the year to the cattle industry. The efficacy of most pathogenic equine parasite. NGS- two parasiticides for controlling horn flies, technology is just starting to gain traction in Corathon® and LongRange®, and their effect the field of clinical veterinary parasitology. on profitability of raising stocker cattle In this study, we tested the ability of NGS for were evaluated. The objectives were to: (1) mass screening of S. vulgaris. Faecal larval determine whether either parasiticide would cultures from 279 horses were collected in a control horn flies better than the control routine diagnostic laboratory receiving equine group, and (2) determine the profitability samples from all over Sweden. In addition to a producer of beef stocker animals could NGS all samples were screened for S. vulgaris expect to achieve by using either of these both by microscopy and amplification of a 171 parasiticides relative to a control group. bp product of the second internal transcribed A total of 302 stocker cattle records from spacer (ITS-2) using standard PCR. For Kansas State University Beef Stocker Unit NGS the ITS2-region was amplified using were analyzed. Horn fly populations were nematode universal NC1 and NC2 primers on average 34% lower with application of flanked by barcoding tags. The resulting Corathon® and 50% lower with application amplicons were prepared and sequenced of LongRange® compared with the control using PacBio RSII SMRT cells. Reads were group. Comparing value of gains with costs, mapped against a local database including all animals receiving the Corathon® treatment nematode rDNA sequences in NCBI. This way gained an average of 10.01 lbs/head (0.1112 lb. the whole strongyle nematode community x 90 days), or 4.55 kg/head, and additional was characterized in each sample. Strongylus profit from the application of Corathon® is vulgaris was identified in 50 (18%) samples estimated at $5.37/head. Animals receiving by microscopy, 62 (22%) by PCR and 30 (11%) the LongRange® treatment gained an average by PacBio. There was substantial agreement of 14.11 lbs/head (0.1568 x 90 days), or 6.41 between microscopy and PCR results (Cohens kg/head, and additional profit from the Kappa 0.7 ±0.05), whereas it was moderate application of LongRange® was estimated a between microscopy and NGS (Cohens t$4.52/head. Although use of LongRange® Kappa 0.6 ±0.07). The agreement between achieved a greater reduction of horn flies, PCR and NGS was weak (Cohens Kappa 0.5 and a greater increase in pounds of gain, its ±0.06). The low sensitivity of S. vulgaris using additional profit is similar to that of Corathon® Pacbio was common especially in samples because of the higher cost per animal to with low relative levels of S. vulgaris DNA. apply (Corathon® = $4.47 per head versus This indicates a negative bias for S. vulgaris LongRange® = $10.00 per head). These by the NGS method. Thus, a low relative profits will be discussed in relation to variable abundance of S. vulgaris DNA may lead price slides in the cattle market. to underestimation of the infection when diagnosed by NGS. Therefore, diagnosis using microscopy or PCR is advised.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 237 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.08 Study on Survival of Nonstrangulating in cases with NSII in order to improve the Intestinal Infarctions Associated With survival rate. Strongylus Vulgaris: Treatment Outcomes of 39 Horses (2008–2019) PS02.09 Pilot Study on the Parasites of Feral Dr. Marie Louise Honoré Jørgensen1, Dr. Horses on Cumberland Island, Georgia, USA Martin K. Nielsen2, Dr. Susanne Nautrup Olsen1, Dr. Páll S. Leifsson1, Prof. Stine Jacobsen1, Dr. Polly Wedlon1,2, Jennifer Ketzis1 Tina H. Pihl1 1Ross University School of Veterinary Medicine, 1University of Copenhagen, Taastrup, Basseterre, Saint Kitts and Nevis, 2Florida State Danmark, 2University of Kentucky, Lexington, University, Tallahassee, United States United States Strongylus vulgaris is the most pathogenic Cumberland Island (147.37 km2), a National gastrointestinal parasite in the horse and Park since 1972, off the coast of Georgia has it is reemerging, in amongst other places, had feral horses for many years, descended Denmark. If nonstrangulating intestinal from horses brought to the island from the infarctions (NSII) caused by Strongylus 1700s through the mid-1900s. The current vulgaris are not diagnosed in due time, the population, estimated to be 120 to 190, is horse may die from this condition. This could unmanaged and consists of approximately most likely be prevented with an increased 15 not well define herds. Health of the horses awareness and recognition of the disease and is a major concern with parasites blamed the initiation of the correct treatment. for many deaths; however, no studies on parasite burdens have been performed and The objective was to investigate the the applicability of data from other island survival rate in horses with NSII undergoing feral horses (e.g., Sable Island off of the coast exploratory laparotomy. of Canada) is uncertain due to differences in island size, climate and horse breeds. During The study is conducted as a retrospective the summer of 2015, with permission from case series. the National Park Service, fresh fecal samples from 15 horses and 6 herds (approx. 10% of the NSII was diagnosed in 39 horses with a horses and 40% of the herds), representing localized intestinal infarction and concurrent various ages, were analyzed using the Cornell- signs of Strongylus vulgaris migration and no McMaster technique. Body condition (BC) signs of intestinal strangulation or enterocolitis. using the Henneke System (scale 1 to 9) for the Data were obtained from medical records in sampled horses was assessed at a distance. the period from 2008 to 2019. Strongyle (15 of 15 positive), Oxyuris (7/15) and Parascaris (6/15) eggs were identified 39 horses were diagnosed with NSII with mean fecal egg counts per gram of 1317 associated with Strongylus vulgaris migration. (225-2525), 86.7 (0-300) and 38.3 (0-300), Exploratory laparotomy was undertaken in 26 respectively. No correlation between BC (mean horses. 12 of these 26 horses were euthanized 4.9; 3 to 7) and strongyle egg counts was intraoperatively due to a poor prognosis. found. The results of this pilot study do not Surgical treatment was carried out in 14 of the indicate that parasites are the primary cause 26 horses and of these 7 (50 %) survived to of health issues with strongyle counts lower discharge. 13 horses were treated medically than that seen in other feral horse populations. due to the owners declining surgery; of these However, interpretation of the study results 100 % were euthanized due to no sign of must be made with caution given that only improvement with medical treatment. a small number of horses were tested and single samples at one time during the year A small group of subjects is the main were analyzed. Further studies are required limitation. to understand factors that influence parasite burdens in these island feral horse populations In conclusion the findings indicate that an and to assist in developing appropriate exploratory laparotomy seems to be justified management plans.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 238 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.10 In Vitro Assays to Evaluate Lethal to be highly active against S. scabiei. A few or Repulsive Activity of Chemical Products products also displayed a repulsive activity Against Sarcoptes Scabiei against the mites.

Dr. Raluca Mindru2, Charbel Al Khoury3, Dr. Fang Fang4, Dr. Charlotte Bernigaud5, Prof. PS02.11 Anthelmintic Resistance in Jacques Guillot1 Gastrointestinal Nematodes of French 1Veterinary College of Alfort, Maisons-alfort, Angora Goat Flocks France, 2Faculty of Veterinary Medicine, Iasi, Romania, 3Holy Spirit University of Kaslik, Dr Christophe Chartier1, Dr Romain Beirut, Lebabon, 4Faculty of Veterinary Cassaigne2, Dr Carine Paraud3 Medicine, Nanning, China, 5Henri Mondor 1BIOEPAR, INRA, Oniris, Nantes, France, Hospital, France 2ANECA, Saint-Ange-et-Torçay, France, 3Anses Niort Laboratory, Niort, France The life cycle of Sarcoptes scabiei does not include an environmental stage per se and Anthelmintic resistance (AR) in transmission may depend on the ability of gastrointestinal nematodes (GIN) is very mites to survive off the host, and to maintain frequent in dairy goats, especially to infectivity. We recently developed in vitro benzimidazoles. Considering that the use of tests to assess the activity of chemical anthelmintics can be quite different between products against motile stages of S. scabiei. dairy and fiber goats, an anthelmintic Mites were collected from experimentally resistance survey was conducted in 15 infected pigs as described by Mounsey et al. French Angora flocks located all over the (2010) and further optimized by Bernigaud country. On each farm, goats were randomly (2016, 2018). Crusts present in the external allocated into several groups of animals: an ear canal were gently removed and placed untreated control group, a group that was in a Petri dish. Adults, nymphs and larvae orally administered oxfendazole (10 mg/ were picked with a needle and used for the kg body weight-BW-) and a group that tests. To evaluate lethal activity, exposure received orally a moxidectin drench (0.4 mg/ to the products was obtained through kg BW). Individual faecal egg counts and immersion, fumigation or direct contact to a pooled larval cultures were done 14 days solid medium (supplemented with a specific after anthelmintic treatment. AR was present concentration of product) according to Brimer when the % of Faecal Egg Count Reduction et al. (1993, 1995). Mites were inspected under (%FECR) was <95 % and the lower limit of the a stereomicroscope after 10 min, 30 min, 1h, 95% confidence interval <90 %. Two farms 2h, 3h, 4h, 8h and 24h. Persistent immobility, were excluded from the analysis because the even when stimulated with a needle was control group was absent. For the 12 flocks considered as death. To evaluate the where oxfendazole was tested, faecal egg repulsive activity, each chemical product was count reductions ranged from zero to 83%, incorporated in Columbia agar through a small meaning that resistance to benzimidazoles hole (3 mm in diameter) or a small piece of was present in all flocks. Post-treatment tissue. The mites were placed in the center larval cultures indicated that Teladorsagia/ of the Petri dish and the migration ability Trichostrongylus was the predominant larval and potential repulsive effect were evaluated type. In the 11 flocks where moxidectin was according to the following criteria defined by tested, resistance was present in 3 of them Brimer et al. (1993): (i) the number of mites with FECR ranging from 69 to 94%, with having demonstrated the ability to migrate, as Teladorsagia/Trichostrongylus being also the indicated by footprints and bacterial colonies, predominant post-treatment culture larval and (ii) the length of the migration tracks. type. This study confirms the extremely high Lethal and repulsive activity was evaluated for prevalence of resistance to benzimidazoles commercially available insecticide/acaricide for GIN in French Angora goats and products (mainly pyrethroids), repellents demonstrates that resistance to moxidectin is (DEET, icaridin, IR3535) but also for several likely developing. essential oils. Several molecules were proved

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 239 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.12 Use of a PCR/Sequencing Technique samples and Haemonchus sequences from for Identifying Haemonchus Species and Genbank indicated multiple genotypes of H. Genotypes From Trichostrongyle Egg DNA contortus and H. placei are present in South Isolated From American Plains Bison (Bison Dakota bison. Bison Bison) Fecal Samples

John McKenzie1, Pratisksha Khanal1, Priscilla S. PS02.13 Anthelmintic and Anti-inflammatory Heliso1, Dr. Michael B. Hildreth1 Activities of Selected Southern African 1South Dakota State University, Brookings, Plants Crude Extracts and Isolated United States Compounds

Haemonchus contortus is a common Mr. Moise Ondua1, Dr. Emmanuel Mfotie Njoya1, trichostrongyle parasite of sheep and goats, Dr. Muna Ali Abdalla1, Prof. Lyndy McGaw1 while H. placei is more commonly found in 1Phytomedicine Programme, Department of cattle. Both species have been identified in Paraclinical Sciences, Faculty of Veterinary Canadian bison, based upon nemabiome Science, University of Pretoria, Pretoria, South metabarcoding of trichostrongyle eggs. The Africa second internal transcribed spacer (ITS2) region of ribosomal DNA has been used to Haemonchus contortus is responsible for over identify genotypes of adult Haemonchus spp. 80% of global parasite infestations of livestock isolated from various ruminants. The purpose and develop in the animal abomasum, causing of this study was to develop a convenient inflammation, anemia, and death. This study was method for species-level identification/ aimed to investigate the biological activities of genotype documentation of Haemonchus eleven southern African plants used traditionally spp. based upon mixed populations of to treat inflammation. Typha capensis, Ficus trichostrongyle eggs isolated from bison elastica, Carpobrotus edulis, Cotyledon orbiculata, fecal samples. For this method, six PCR and Senna italica crude extracts showed high primer pairs were evaluated for their ability antioxidant (IC50 range between 0.22 g/mL to consistently amplify a major portion of ± 1.18 and 7.11 g/mL ±1 .71), anti-inflammatory the Haemonchus spp. ITS2 region so that using 15- lipoxygenase inhibition (IC50 µrange the amplified product could be sequenced. between 3.47µ g/mL ±0.07 and 18.00 g/mL The goal was to identify two primer pairs ± 4.23), and anthelmintics against nematode collectively spanning the entire ITS2 region, parasites H. contortusµ and free-living nematodesµ with each pair consisting of a genus-specific Caenorhabditis elegans with LC50 between primer and an ITS2 universal primer. Samples 184.94 g/mL ± 2.62 and 2669.67 g/mL ± 4.24. tested included adult and larval Haemonchus Two new compounds were isolated for the first spp. and mixed trichostrongyle egg samples time fromµ T. capensis (Isorhamnetin-3-O-µ -D- from sheep, cattle, and bison; the analyses glucoside, and Isorhamnetin 3-O-rutinoside) included amplification consistency, quality showed good anti-inflammatory and anthelminticβ values (QV) from Sanger sequencing, and activities. Typha capensis showed good activity sequence fidelity between individual worms not only against H. contortus larvae but by and mixed egg samples. A primer pair set inhibition of inflammatory mediators, increasing composed of a forward universal primer in consequently the healing process on the the 5.8S subunit gene coupled with a genus- abomasum of the animals. Typha capensis can specific reverse primer (at the ITS2 3’ end, be used as potential source of new remedies extending into the 28S gene) provided high against inflammation and nematodes infections. QV values over the ITS2 region, except for There was high correlation between antioxidant the 3’ end. Two other primer sets provided and anthelmintic activities, suggesting that high QV values in the 3’ half of ITS2. Egg antioxidant assay can be used as bioassay-guided DNA results were consistent with individual fractionation for isolation of compounds with worm results. Haemonchus placei and H. anthelmintic activities. contortus were found in cattle samples; only H. contortus was found in sheep samples. *Corresponding author: [email protected] MEGA7 generated phylogenies from bison Tel.: (+27) 748308068

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 240 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.14 Effect of Trichostrongylus infection could negatively impact the nitrogen Colubriformis Infection and Phosphorus metabolism, increasing the nitrogen losses via Dietary Levels on Lamb’s Nitrogen Metabolism urine.

Ms. Thiago F V Bompadre1, Dr. Gabriel Z Sakita1, Mr. Victor M Nakaguchi1, Ms. Isabella P PS02.15 Epidemiological and Molecular Moraes1, Prof. Alessandro F T Amarante2, Prof. Updates on Cystic Echinococcosis in Wild Adibe L Abdalla1, Prof. Helder Louvandini1 Boars from Italy 1University of São Paulo, Piracicaba, Brazil, 2Sao Paulo State University, Botucatu, Brasil Dr. Giovanni Sgroi1, Dr., PhD Antonio Varcasia2, Dr. Giorgia Dessi2, Dr. Nicola D’Alessio3, Dr. The aim of the trial was to compare the Mario Santoro3, Dr. Claudia Tamponi2, Dr. nitrogen metabolism of growing lambs fed Urmas Saarma4, Dr. Vincenzo Caputo5, Dr. with different phosphorus dieraty levels Paolo Sarnelli6, Dr. Valerio Toscano5, Dr. Paolo and infected with T. colubriformis. Eighteen Varuzza1, Dr. Antonio Scala2, Dr. Alessandro Santa Ines castrated male lambs (31.8 ± 4.58 Fioretti1, Dr. Vincenzo Veneziano1 kg of body weight), averaging 6 months of 1Department of Veterinary Medicine and age were randomly allocated in a factorial Animal Productions, University of Naples arrangement 2 × 2. The factors were 2 Federico II, Napoli, Italy, 2Department of diets containing 50% (Low-P diet) or 100% Veterinary Medicine, University of Sassari, (Adequate-P diet) of their P requirements Sassari, Italy, 3Istituto Zooprofilattico based on the National Research Council, Sperimentale del Mezzogiorno, Portici, Italy, and two infection status (infected or not). 4Department of Zoology, Institute of Ecology The infected lambs received a single oral and Earth Sciences, University of Tartu, dose of 40,000 L3 of T. colubriformis. The Tartu, Estonia, 5Regional Reference Center of animals were individually housed in stalls Urban Veterinary Hygiene (CRIUV), Napoli, for a 96-days experimental period. The Italy, 6UOD Prevenzione e Sanità Pubblica diets were formulated with a forage source Veterinaria, Regione Campania, Napoli, Italy (60%) of Tifton 85 bermudagrass hay (Cynodon dactylon cv.), and a concentrate Cystic echinococcosis (CE) is one of the source (40%) composed by cassava meal most important worldwide parasitic zoonotic and glutenose, and the adequate-P diets disease and it represents an important public was also supplemented with dicalcium health and socio-economic concern. In the phosphate. This diet composition provided last years, the wild boar in Italy is increased 14.7% of crude protein, 0.38%, and 0.18% and this ungulate could play an important of phosphorus for adequate-P and low-P role in the spreading of CE in wildlife. diets, respectively. Thirty-six days after This survey was carried out to determine infection, an apparent digestibility trial was prevalence of hydatid cysts and genotypes performed, where the feed intake, feces of E. granulosus circulating in the wild boar and urine were weighed and sampled for population from Campania Region, Southern nitrogen analyses. The Kjeldahl laboratorial Italy. The carcasses of animals, obtained from technique was used to determine the sample different boars hunting areas (BHAs), were nitrogen concentration. The N-absorption examined during two hunting seasons (2016- was calculated based on N-intake minus 2017) by 51 veterinary involved in the project N-feces, and N-retention was caclulated as Piano Emergenza Cinghiali Campania. Wild N-absorbed minus N-urine. All statistics were boar origin, gender and age were collected performend using the R software, and the per each animal. When cysts were found, fixed effects and interactions were compared their number, morphology and fertility by Tukey’s test (P<0.05). The results indicated were determined by visual and microscopic that the infection and the P dietary did examination. Cysts were classified as fertile, not affect the N-absorption. However, the sterile, caseous and calcified. Protoscoleces N-retention was greater in non-infected (46% and germinal layers were collected from of N-intake) compared to infected (29% of individual cyst and DNA was extracted. A N-intake; P<0.01) animals. In conclusion, the specific molecular diagnosis was obtained

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 241 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

by sequencing PCR-amplified mitochondrial of this study it can be concluded that CE is genes encoding for the NADH dehydrogenase not an issue in the Gambian abattoirs and subunit 4 (ND4), ATPase subunit 6 (ATP6), no financial losses are incurred as a result of NADH dehydrogenase subunit 2 (ND2) and organ condemnation. partial cytochrome c oxidase subunit 1 (COI). Out of a total of 2,108 wild boars examined For a better understanding of the CE status for CE, 93 (4.4%) were found positive. The in The Gambia, further investigations should total number of cysts collected was 123, of be carried out using advanced molecular which 118 (95.9%) in the liver, 4 (3.3%) in the diagnostic techniques on samples from both lungs and 1 (0.8%) in the spleen. Cysts were the definitive and intermediate hosts to rule 70 (56.9%) fertile and 53 (43.1%) sterile/ out the existence of the parasite in this country. acephalous. The presence of fertile cysts was detected in 19,4% of positive boars. Out of a total of 28 boars, molecular diagnosis showed PS02.17 Molecular Identification and 19 (67.9%) of these infected with the pig strain Function Analysis of SUMO and UBC9 Genes (G7). These results document the prevalence of Taenia Pisiformis of E. granulosus infection and reveal, for the first time, the presence of the pig strain (G7) Dr. Shaohua Zhang1, Dr. Xuenong Luo1, Dr. in wild boars from Italy. Aijiang Guo1, Dr. Shuai Wang1, Dr. Xuepeng* Cai1 1State Key Laboratory of Veterinary Etiological PS02.16 Prevalence and Economic Losses of Biology, Lanzhou Veterinary Research Cystic Echinococcosis in The Gambia Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China Mr. Saidal Ali Bah1, Dr. Suzanne Gokool2, Prof. Volkan Akyol1 SUMOylation is an essential post-translational 1Uludag University, Nilufer, Turkey, 2Northwick modification for protein in eukaryotes, which Park Institute for Medical Research Tropical participates in protein-protein interactions, Parasitic Diseases, United Kingdom subcellular localization, protein stabilization and diverse cellular process regulation. The ABSTRACT SUMOylation pathway has been studied in Cystic Echinococcosis (CE) is globally several model species. However, SUMOylation distributed, is super endemic in East and system and processes in tapeworms remain North Africa and causes great financial losses unknown. In the present work, the full- to the livestock industry in these regions. In length cDNA of two core components genes, West Africa, the disease is suspected to be small ubiquitin-like modifier (TpSUMO) and prevalent but few epidemiological studies E2-conjugating enzyme (TpUBC9), in the have been conducted so far. The aim of this SUMOylation pathway of Taenia pisiformis study was to determine the presence of the were obtained through rapid amplification disease and estimate the prevalence of CE in of cDNA ends using the polymerase chain slaughtered livestock in The Gambia. reaction (RACE-PCR) strategy. Bioinformatic In five abattoirs located across The Gambia, analysis showed that the TpSUMO gene from July to December 2017, a total number contained a 309 bp ORF, encoding 103 amino of 1968 slaughtered cattle, sheep and goats acids with a predicted weight of 11.4 kDa and were inspected for the presence of CE using the TpUBC9 gene with 516 bp ORF encoding a ante-mortem and post-mortem examinations. 172 amino acid protein with a putative mass of The distribution of the animals examined was 19.6 kDa. The His-TpSUMO (~18 kDa) and His- 56.1% goats, 28.9% cattle and 15.0% sheep. TpUBC9 (~25 kDa) proteins were expressed in The average age of the animals slaughtered Escherichia coli Transetta (DE3), respectively. was 4-6 years old. Of the total number of In addition, using leucine aminopeptidase of examined animals, no hydatid cysts were Taenia pisiformis (TpLAP) as a SUMOylation identified 0/1968 (0.0%) and 16/1400 (1.1%) substrate we further demonstrated the calcified cysts were collected from the total physical interaction of TpLAP-TpUBC9 number of sheep and goats. From the result via one-to-one yeast two-hybrid and co-

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 242 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

immunoprecipitation analyses. The truncated PS02.19 Do Dirofilaria Immitis Infections N-terminus (aa 1-185) and C-terminus (aa in Aedes Aegypti Affect Wing Beat 186-522) of TpLAP interacted with TpUBC9, Frequencies? respectively, while TpLAP-TpSUMO or TpLAP- TpSUMO GG showed no direct interactions, Doyeon Park1, Haripriya Mukundurajan2, Joy suggesting that the SUMOylation of TpLAP Bowles3, Haluk Kalin1, Dr. Manu Prakash2, Dr. was mediated△ by TpUBC9. The findings Lindsay Starkey3, Dr. Byron Blagburn3, Dr. provide solid evidence that the SUMO Sarah Zohdy1,3 modification pathway plays an important part 1School of Forestry and Wildlife Sciences, in regulating the protein functions involved in Auburn University, Auburn, United States, adult worm growth and development. 2Department of Bioengineering, Stanford University, Stanford, United States, 3Department of Pathobiology, College of PS02.18 Field Flea Collections Reveal Veterinary Medicine, Auburn University, New Host and Distribution Records for Auburn, United States Stenoponia Americana (Baker, 1899) and Amphipsylla Washingtona (Hubbard, Dirofilaria immitis, the causative agent of 1954) (Siphonaptera: Ctenophthalmidae, canine heartworm disease, is transmitted by Leptopsyllidae) in the United States numerous genera and species of mosquitoes. Principally among them are Aedes, Dr. Gebbiena Bron1, Dr. Michael Hastriter2 Anopheles, Culex, and Ochlerotatus spp. 1University Of Wisconsin - Madison, Madison, Within the mosquito, ingested microfilariae United States, 2Monte L. Bean Life Science develop through three developmental stages Museum, Provo, United States (L1, L2, L3). A recent study demonstrated that mobile phones are capable of capturing In the United States, fleas (Siphonaptera) acoustic data from mosquito wingbeats. were extensively collected and described Since each mosquito species has a different in the 20th century, but currently field wingbeat frequency by which they attract collections are rare and primarily aim to mates, a brief wingbeat recording (<1/10th further understand their role in plague disease of a second) can be analyzed to determine ecology. In this study, fleas were collected mosquito species and thus its capability to from small rodents at several locations in transmit disease. Here, we examine wingbeat the United States during plague disease signatures and flight duration patterns of ecology studies in the summers of 2013 Aedes aegypti to determine if wingbeat to 2015. Twenty specimens of Stenoponia mobile phone recordings can be used americana (Baker, 1899) (Siphonaptera: to distinguish infected and non-infected Ctenophthalmidae) collected from sagebrush mosquitoes. Individual female mosquitoes voles (Lemmiscus curtatus) and deermice were placed into a chamber at various time (Peromyscus maniculatus) on the Awapa points after feeding on normal blood or blood plateau (Wayne County, Utah) are an containing microfilariae and recorded with expansion of the geographical range and a mobile phone for 60 seconds. Recordings host distribution of this large flea species. were processing using an in-house Python Amphipsylla washingtona also collected script to determine wingbeat frequency and from Lemmiscus curtatus, Wayne County, flight duration. One hundred sixty recordings Utah is a new record for Wayne County and were obtained. The presence and number expands the known distribution for this rare of heartworm larvae were determined by species an additional 600 kilometers from mosquito dissection. Our findings indicate additional records published herein from the that L3 (infective stage) infected mosquitoes BYU flea collection from the National Reactor have significantly lower (mean=429 Hz) Testing Station near Idaho Falls, Butte County, wingbeat frequencies than age-matched Idaho. These observations illustrate that negative mosquitoes (mean= 577 Hz; flea distributions are still changing and that p<0.0001). Flight duration based on wingbeat trusted handbooks and regional identification recordings was substantially lower (17.6% keys need temporary updating. of the time) in L3 infected mosquitoes than

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 243 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

mosquitoes fed on negative blood (35.9% of anthelmintic treatment. These results indicate the time). We present data suggesting that that CarLA® is unaffected by anthelmintic wingbeat frequencies may be used to identify treatment, and that levels measured during a mosquitoes infected with pathogens of public lamb’s first grazing season are predictive of health or veterinary concern. subsequent levels. Therefore, under Ontario grazing conditions, selection of replacement ewes with elevated CarLA® may reduce PS02.20 Correlation of Salivary Antibody pasture contamination following lambing and to Carbohydrate Larval Antigen With during their second grazing season. Gastrointestinal Nematode Parasitism in Sheep Under Ontario Grazing Conditions PS02.21 Developmental Activation of Emma Borkowski1, Niel Karrow1, Paula Infective Strongyloides Stercoralis Larvae Menzies1, Jacob Avula1, Brandon Lillie1, Andrew After Percutaneous Migration Through an In Peregrine1 Vitro Model 1University Of Guelph, Guelph, Canada Ms. Shannon Kitchen1, Dr. Tegegn Jaleta1, Dr. Gastrointestinal nematodes (GINs) are an Zhu Wang2, Dr. David Mangelsdorf2, Dr. James important cause of morbidity, mortality Lok1 and financial loss on Ontario sheep 1University Of Pennsylvania, Philadelphia, farms. In light of the rising prevalence of United States, 2University of Texas anthelmintic resistance, selecting sheep Southwestern Medical Center, Dallas, United with a superior immune response to GINs States is an attractive complementary control strategy. In New Zealand, such sheep are Strongyloides stercoralis is a global identified by measuring salivary antibody parasitic nematode of humans and canids. to a carbohydrate larval antigen (CarLA®, While infections are usually self-limiting, AgResearch Inc.). However, GIN epidemiology hyperinfection syndrome can occur in under cold continental grazing conditions, immunocompromised individuals, in which such as in Ontario, is different from New autoinfective larvae disseminate throughout Zealand. The purpose of this study was to the body and helminth burden reaches fatal determine whether salivary CarLA® antibody levels. Currently, albendazole and ivermectin correlates with GIN burden under Ontario are the only drugs deemed effective against grazing conditions. Replacement ewe lambs the disease, but often result in incomplete (n=107) on an Ontario commercial sheep eradication due to the nematode’s unique farm were followed for two years, including autoinfective lifecycle. Consequently, their first lambing and lactation. GIN fecal understanding larval developmental pathways egg counts (FECs) were monitored every 6-8 within the lifecycle is necessary for progress weeks from May-November in 2016 and 2017. towards identifying new therapeutic agents. Salivary CarLA® antibody was measured at Parallels between S. stercoralis infective the beginning (May), middle (August) and third-stage larvae (iL3) arrest/reactivation end (November) of each grazing season, and and the dauer entry/recovery mechanisms in at mid-gestation in late winter (March). Mean Caenorhabditis elegans have been identified, CarLA® levels increased in 2016, declined including the importance of steroid hormones over winter, and rapidly increased during the known as dafachronic acids (DA) in binding 2017 grazing season. Spearman correlation to the nuclear hormone receptor DAF-12 to coefficients between CarLA® levels were initiate dauer exit. Although the mechanism consistently positive, of weak to moderate for DA production is known, which involves strength (rho = 0.19-0.58), and generally a cytochrome P450 enzyme (CYP), the significant (p = 7.3 X e-11 to 4.7 X e-2). In a exact cue to begin synthesis of DA is not. To multivariate mixed model, increased salivary examine this, we utilized in vitro percutaneous CarLA® was significantly associated (p <1.0 larval migration chambers with mouse skin X e-4) with decreased GIN FEC throughout and a culture medium that does not permit both grazing seasons, regardless of previous spontaneous resumption of feeding upon a

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 244 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

shift to host-like temperature. In cohorts of iL3 for H. contortus, and 31 days for H. placei. 42 that penetrated mouse skin, 51.7% resumed days after the artificial infections, the 14 kids feeding, as reflected in fluorescein ingestion, were slaughtered, and the Haemonchus sp. while only 9.6% of non-penetrating controls parasites were harvested, in totum, from the did so. This iL3 activation via skin migration abomasum. The experimental groups GI, GII, was further investigated using LC/MS to and GIII had, respectively, an average of 25.5, determine whether there was a correlation 619.5, and 724.75 (120 H. placei, and 604.75 H. with DA production, however preliminary contortus) adult specimens, and no immature results did not indicate a peak in synthesis. forms. Under the conditions of this study, the Considering the possibility that DA levels viability of goat infection by H. placei was required for resumption of feeding are below confirmed, although, with low susceptibility. the level of LC/MS detection, we are now Nevertheless, the parasitism of this helminth assessing the effect of ketoconazole, a CYP species was more intense when associated inhibitor, to ascertain the requirement of CYP with H. contortus. This fact indicates that in function for DA biosynthesis in penetration- common grazing between cattle and young mediated developmental activation of S. goats, when the latter end up ingesting both stercoralis iL3. Moreover, genetic analysis is Haemonhcus species, especially in a mixed being performed on skin-penetrated larvae to infection, H. placei may also parasitize them. identify transcripts associated with the dauer recovery pathway. PS02.23 Ovicidal Effect of Citronellal and Citronellol in Haemonchus Contortus In Vitro PS02.22 Viability of Haemonchus Placei Egg Hatch Test Parasitism in Experimentally Infected Young Goats Poliana Pacheco1, Jessica Ribeiro2, Bruna Wedy2, Danielle Balbino2, Guilherme Luciani2, Dr. Isabella Santos1, Dr. Willian Maciel1, Dr. Karen Akashi3, Dr. Rodrigo Giglioti2, Cristiane João Toscano1, Dr. Gustavo Felippelli1, Dr. Fiorin2, Leandro Rodrigues2, Bianca Azevedo2, Breno Cruz1, Dr. Ana Chagas4, Dr. Vando Gunta Gutmanis2, Dr. Cecilia Veríssimo2, Dr. Soares3, Dr. Lucas Gomes1, Dr. Welber Lopes2, Helder Louvandini1, Dr. Luciana Katiki2 Dr. Alvimar Costa1, Dr. Gilson Oliveira1 1University Of São Paulo, Piracicaba, Brazil, 1Universidade Estadual Paulista - UNESP, 2Institute of Animal Science (IZ/ APTA/ SAA), Jaboticabal, Brazil, 2Universidade Federal de Nova Odessa, Brazil, 3University of State of Goiás - UFG, Goiânia, Brazil, 3Universidade Maringá, Umuarama, Brazil Brasil - UB, Descalvado, Brazil, 4Embrapa Pecuária Sudeste, Empresa Brasileira de Gastrointestinal nematodes are one of the Pesquisa Agropecuária - EMBRAPA, São main causes of decrease in the productivity Carlos, Brazil of small ruminants and are commonly treated with anthelmintics. However, the The present study aimed to evaluate the indiscriminate use of these drugs led to viability of Haemonchus placei parasitism the reduction of efficacy through the in experimentally infected goats, since selection of resistant parasites, so it is there are no such studies regarding this essential to preserve the efficiency of the ruminant species. For that, 14 newborn male current anthelmintics, exploring new control Saanen kids were placed in one of the four alternatives such as the use of phytochemical experimental groups: GI – infected with 5000 compounds as essential oils. The objective of H. placei L3 (n = 4); GII – infected with 5000 the present study was to evaluate the activity H. placei L3 (n = 4); GIII – infected with 2500 of citronellal and citronellol on Haemonchus H. contortus L3 + 2500 H. placei L3 (n = 4), contortus resistant strain through the egg and GIV – control, inoculated with distilled hatchability test and to determine lethal water (n = 2). Each kid received, orally, the concentrations CL50 and CL90 using infective dose in a single inoculum. Based increasing doses (0.003 mg / mL, 0.007 on daily fecal egg counts (FEC), the average mg / mL, 0.010 mg / mL, 0.030 mg / mL, pre-patent period was determined as 24 days 0.060 mg / mL, 0.120 mg / mL, 0.250 mg

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 245 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

/ mL and 0.500 mg / mL). The eggs were eggs used in the tests were recovered from placed in 48-well plates and incubated with adult donor sheep. Egg hatch tests (EHT) the described concentrations for 24 hours at were performed with serial concentrations 27°C. The hatchability was evaluated under from 1.0 mg/mL to 0.004 mg/mL to obtain the inverted microscope. The calculation a dose-response curve and reach the 99% of lethal concentrations CL50 and CL90 effective concentration. EHT results for the were performed through the SAS Probit resistant isolate showed 99.5% efficacy at program, with the independent variables concentrations above 0.5 mg/mL. However, (dose) transformed by natural logarithm the EHT results for the sensitive isolate were (log dose). Citronellol presented the best much lower at 32.3% at 0.5 mg/mL and anthelmintic efficacy when compared to 41.8% efficacy at the maximum concentration citronellal as LC50 was 0.02 and Citronellol tested (1.0 mg/mL). Our results suggest 0.63 mg / mL, and CL90 was 0.63 and 3.26 that the components in Mentha piperita mg / mL, respectively. It was concluded essential oil probably have a different mode that the use of essential oils compounds of action in gastrointestinal nematodes as had a positive influence on H. contortus the resistance background to benzimidazole eggs, especially citronellol. The low dose was not associated with EHT results for the of Citronellol to inhibit the hatch of eggs studied isolates. Thus, the importance of reinforces the potential anthelmintic activity testing natural compounds on more than present in essential oil compounds and one nematode population is evident in order deserves further scientific investigations. - to provide a better overview of the actual FAPESP/2018-02423-0 efficacy of said compounds.

PS02.24 Efficacy Variation of Mentha PS02.25 Encysted Cyathostomin Larval Piperita Essential Oil on Haemonchus Counts: Mucosal Digestion Revisited Contortus Isolates With Differing Benzidazole Resistance Backgrounds Haley Zynda1, Dr. Martin Nielsen1, Jessica Kenealy1, Dr. Ashley Steuer1, Haley Anderson1 Dr. Marcel Teixeira1,2, Dr. Fernando Raul 1University Of Kentucky, Lexington, United Correia de Vasconcelos2, Dr. Janaelia Ferreira States Vasconcelos2, Dr. Jomar Patricio Monteiro1,2, Dr. Luiz Silva Vieira1 Cyathostomins are pervasive equine parasites 1Embrapa Caprinos e Ovinos, Sobral, Brazil, and may cause larval cyathostominosis. Upon 2Centro Universitário UNINTA, Sobral, Brazil ingestion of the third larval stage, the larvae encyst in the mucosal membranes of the The spread of anthelmintic resistance made cecum, ventral colon, and dorsal colon. Once treatment options against gastrointestinal encysted, they can arrest development and parasites increasingly scarce so the search for accumulate in high numbers. No published natural compounds as antiparasitic candidates study has investigated spatial variation is now an important field. However, because of encysted larvae within each intestinal concentrations of active compounds may vary compartment and the current mucosal according to their origin and post-harvest digestion protocol lacks a description of a processing, there are many obstacles for the standardized area from which to take the establishment of standards hindering reliable tissue sample. Therefore, this study sought results. To date it is not known if a given to evaluate spatial variation in encysted compound may vary on efficacy between larval counts in defined sections of each parasite populations. In this work we evaluate large intestinal organ. Following humane the efficacy of Mentha piperita essential oil euthanasia, ceca, ventral, and dorsal colons against eggs of two Haemonchus contortus were harvested from 8 foals raised in an isolates with and without history of resistance anthelmintic naïve parasitology research to benzimidazole. Tests were carried out herd. Each organ was weighed and separated using the standard egg hatch inhibition test into 3 equal sections by length: the orad, with a 2% Tween 80 as diluent. Nematode intermediate, and aborad portions. From

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 246 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

each of those sections, two-5% weight population per field of vision on the average tissue samples were selected and underwent was 2.9±0.43 in the control group; 8.5±2.3 in mucosal digestion to quantify the number the first test group; 9.4±0.71 in the second of early third stage larvae (EL3) and late test group. On the 15th day of storage, the third stage larvae/fourth stage larvae (LL3/ bacterial count in 1 g of meat in the first test L4). A mixed model statistical analysis was group was (43±13.4)•103; staphylococcus - performed to evaluate for differences of larval (57±26.2)•103; streptococcus - (9.4 ±2.7)•103; counts between the different organs, sections, in the second test group respectively - and the interaction terms between the (139±13.4)•103, (75.3±26.2)•103, (11.0±2.7)•103; organs and sections. There were significant and the control group: (2.6±0.4)•103, differences between organs (P = <0.0001), (1.2±0.8)•103 and (1.4±0.6)•103. with the cecum having higher counts than the ventral and dorsal colons. However, there The mixed invasions affect the sanitary quality were no significant differences between the and food value of the products received, and three defined sections (P = 0.1076). Although makes meat less stable upon storage. The meat not significant, the following sections had the is fresh during 24 hours after the slaughter; on highest average counts of encysted larvae: the 15th day its bacterial content increases and intermediate cecum, orad ventral colon, shows signs of doubtful freshness. and aborad dorsal colon. These trends may become significant in larger data sets and should therefore be investigated further in PS02.27 Parasitic Structures Carried by future studies. Sarcophagidae Fly in Two Different Biotopes in the Region of Teodoro Sampaio, Sao Paulo, Brazil PS02.26 Veterinary-Sanitary Evaluation of Mutton with Mixed Invasions Karen Akashi1, Dr. Leonice Dias2, Jessica Paz1, Leandro Rodrigues4, Poliana Pacheco4, Dr. Dr. Shermatov Sagynbek1 Rogerio Giuffrida3, Dr. Cecilia Verissimo4, Dr. 1Naryn State University, Naryn, Kyrgyzstan Luciana Katiki4, Dr. Vamilton Santarém3 1State University of Maringá, Umuarama, The aim of the research is to examine Brazil, 2State University Paulista, Presidente veterinary-sanitary quality of mutton with Prudente, Brazil, 3University of West Paulista, mixed invasions. The mutton carcasses are Presidente Prudente, Brazil, 4Institute of divided into 3 groups depending on the Zootecnia, Nova Odessa, Brazil intensity of co-members consisted in the Helminthic cenosis. The first group included The synanthropic flies of the Sarcophagidae the material from non-infested animals (Diptera, Insecta) family are potential (control group); the second group (first test mechanical vectors of etiological agents group) included samples from two-members such as virus, bacteria, protozoan cysts and (dicroceliasis + monieziasis); the third group helminth eggs. Sarcophagidae have been (second test group) included materials from implicated in facultative wound myiasis. Poor three-members (dicroceliasis + monieziasis + sanitation is probably the most important strongylatosis). risk factor for human and animal myiasis. The present study aimed to evaluate the The probes of the meat were taken in presence of parasites in Sarcophagidae 24 hours and on the 15th day of storage flies captured in two different areas: a rural (0…4oC) after the slaughter of sheep. The settlement community and in a countryside meat of the animals belonging to the II and city of the Brazilian Southeast. For trapping III groups, unlike that of the control group, of the flies, five traps were used per bait with showed the humidity increase by 5.1 and 7.9 attract of cow liver and fish. Capture of flies % respectively; proteins decreased by 3.6 was carried out at the beginning, middle and and 6.6%; fat - by 27.2 and 30.1%; caloric in the end of each season (March/2012 to value - by 18.3 and 21.1%; In 24 hours after the February/2013), by using of traps. Captured slaughter of helminthic animals, the microbial flies were washed for evaluating the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 247 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

exoskeletal, and desiccated for the removal May 7th, 22 days after infestation, 8 hours of the intestines. The fluid obtained from the after blockage of the ulcer with an adhesive external lavage and intestinal content were tape, a live larva was removed mechanically. separately subjected to the centrifugation- The specimen was identified as an L2 stage flotation and sedimentation methods for of Dermatobia hominis. Ticks do not fly and observation of parasitic structures (helminth have restricted dispersal around the host eggs and protozoan oocysts/cysts). A total after they leave it. These facts make ticks of 88 insects were captured, 53 flies in the poor vectors of D. hominis eggs. Since D. city and 35 in the rural area. The parasite hominis is reported to perform oviposition on structures observed were: Giardia spp. (2 inanimate objects despite of flying diptera, cysts, 2 in the city), Entamoeba spp. (10 cysts, and ticks are hematophagous attaching to 6 in the city and 4 in the rural area) and non- the skin of vertebrate hosts, the rare use sporulated oocysts of protozoa (3 oocysts, 1 of ixodidae ticks as successful egg vectors in the city and 2 in the rural area). There was is possible, as previous described by Dunn a significant difference between the number (1918). An adequate eco-epidemiological of cysts recovered from the external portion situation with high abundance of A. sculptum in relation to the intestine (p <0.05 test t). ticks, D. hominis and different co-existing The results show that Sarcophagidae flies vertebrate hosts might have contributed to can carry parasites potentially pathogenic this exceptional report. to humans and animals, both in the rural and countryside city areas. PS02.29 Diversity of Culicoides Species in New England, NSW, Australia: 28 Years of PS02.28 Amblyomma Sculptum (Acari, Data Ixodidae) as a Probable Vector of Dermatobia Hominis (Diptera, Cuterebridae) Mr. Biniam Tsegaye Lakew1, Prof. Stephen W on a Human Cutaneous Infestation in Brazil Walkden-Brown1, Dr Adrian H. Nicholas2 1University Of New England, Armidale, Andre Cutolo1 Australia, 2Biosecurity & Food Safety NSW, 1Instituto Adolfo Lutz, São Paulo, Brazil Central Coast Primary Industries Centre, Gosford, Australia Dermatobia hominis is a conspicuous Neotropical parasite fly and its larva Culicoides are biting midges approximately parasitizes different species of vertebrate 3mm in length with females feeding on blood hosts including man. On April 15th, 2018 of a wide range of mammalian and avian the author (AAC) visited a rural property hosts. They play an important role as vectors in Itirapina, São Paulo state, Brazil. The site of more than 50 viruses and can cause host had few heads of cattle heavily parasitized annoyance themselves when present in by D. hominis and pasture areas with horses large numbers. In Australia, the distribution keeping a population of Amblyomma of Culicoides is monitored by the National sculptum in the environment. On the visit day Arbovirus Monitoring Program (NAMP). New AAC was wearing boots, trousers and long England is in the north of the state of New sleeved shirts aiming to be protected from South Wales and is an important area of mosquito bites and walked around but with cattle and sheep production. Under the NAMP trousers openings not sealed allowing tick’s surveillance program, Culicoides were trapped infestation. On the same day an Amblyomma from thirteen different locations in this region sculptum adult tick was found on AAC’s during the spring to autumn months over a arm. Two days later AAC, which is allergic to period of 28 years (1990 – 2018) using light tick bites, started to scratch a lesion on the traps set at approximately monthly intervals external face of his left thigh. On April 28th for a period of 2 nights during the new episodes of pain like stings on the lesion moon near cattle camps. Samples from 4482 started to occur. On May 3rd an ulcer less collections (672 in Spring, 2118 in Summer than 1 mm wide was identified and movement and 1692 in Autumn) were enumerated and of the caudal end of a larva was noticed. On speciated. Of these trapping events 85.2%

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 248 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

resulted in successful Culicoides catches with the individual samples that were collected. a total catch of 156,101, a median catch of 4 These NWS markers lay the foundation for a and mean log10 catch of 0.87. A total of 25 SNP database for this species which can be different species were identified, of which used to assess the presence of diagnostic 15 have been reported to feed on humans, 9 SNPs that discriminate by population and can on sheep, 8 on Australian marsupials, 7 on facilitate the identification of the geographic horses, 6 on cattle and smaller numbers on origin of NWS flies associated with future birds, dogs, flying foxes and rabbits. For 7 potential outbreaks. species, no information on the mammalian/ bird host is available. The most trapped 6 species in descending order were C. marksi, PS02.31 First Report of Philornis sp. in a C. austropalpalis, C. victoriae, C. dycei, C. Protected Area in Uruguay bundyensis and C. brevitarsis. The highest mean catch counts were from the plains Mrs. María Ernestina Olhagaray Torres1, (mean altitude 198 m) followed by the slopes Dr. Ana Laura Pérez Sarasqueta1, Dr. César (512 m) and the tablelands (871 m). Counts Echaides Ortíz2, Dr. Eleonor Castro-Janer1 were lowest in spring, highest in summer and 1Departamento de Parasitología, Facultad intermediate in autumn. A diverse range of de Veterinaria, Universidad De La República, Culicoides species is present in the region. Montevideo, Uruguay, 2Intendencia Municipal de Montevideo, Montevideo, Uruguay

PS02.30 New World Screwworm The genus Philornis (Diptera: Muscidae) is (Cochliomyia Hominivorax) Population composed of about 50 species, which are Structure Analysis Using Genotyping by distributed mainly in the neotropical zone. Sequencing The adults of Philornis are characterized by laying their eggs in bird nests. After that, the Dr. Robert Mitchell III1, Dr. Adalberto Pérez larvae penetrates the skin, producing myiasis de León1, Dr. Felix Guerrero1, Dr. Scott Geib2, in the chicks. Due to their subcutaneous Dr. Sheina Sim2, Dr. Agustin Sagel3, Dr. Pamela location and type of feeding they generate Phillips4, Dr. John Welch5 delays in the development of chicks and, in 1USDA-ARS, Kerrville, United States, 2USDA- some cases, death. ARS, Hilo, United States, 3USDA-ARS, Pacora, United States, 4USDA-APHIS, Pacora, United As part of a study of dipterous of Veterinary States, 5USDA-APHIS, College Station, United importance, carried out in the Lecocq Park States Zoo (Montevideo), dipterous samples were taken during 2018. Larvae of the New World screwworm (NWS) fly (Cochliomyia hominivorax) feed on living The Zoo is located inside the Protected tissue of warm-blooded hosts, including Area: St. Lucia Wetlands (86.517 hectares), livestock and humans, causing myiasis. While where conservation plans are carried out for the U.S. was officially declared NWS-free in endangered species. Here, over 150 species 1966 there have been sporadic introductions of vertebrates and invertebrates develop in of NWS into the U.S. including one of the wildlife. Every month specimens of Philornis most recent outbreaks that occurred in the were collected. Florida Keys in September of 2016. To further explore the origin of NWS flies collected Many of the birds present in this Protected during the 2016 outbreak, we applied Area carry out two migratory flows, some Genotyping by Sequencing (GBS) to analyze are winter migrations and others are visitors samples collected from Florida, the Caribbean, of the summer months. Of this last group, Central, and South America to identify several reproduce in the Area. In addition, diagnostic single nucleotide polymorphisms species that come from migratory movements (SNPs) to distinguish population structure. from tropical climates have recently been This method yielded genotypes for thousands found in Uruguay, perhaps due to the effects of loci across the entirety of the genome from of climate change.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 249 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

captured. In contrast, the abundance of both The protection of the species requires species in the dairy farm was higher. A total international work and commitment. of 52,017 insects were captured and 16,444 According to the Red List of birds in Uruguay, were identified as M. domestica and 3,206 as of the 458 taxa evaluated, representing 453 S. calcitrans. species, 46 were identified as Threatened, 31 Vulnerable, 12 Endangered and two Critically In both locals, the fly fluctuation was similar. Endangered. An increase in the M. domestica population was observed in May 2018. In the case of This communication represents the first the stable fly, an increase in the population report of Philornis sp. in a Protected Area in was observed during spring (November- Uruguay, which is a very important alert to the December), and a little peak in winter (June) conservation of bird species because they could was registered. The winter in this year was be affected by the presence of this parasite. extremely warm.

In the Zoo, other little peak of S. calcitrans PS02.32 Seasonal Abundance of Stomoxys population was observed in autumn, during Calcitrans and Musca Domestica in the South March- April 2018. Apparently, the behavior of Region of Uruguay (2018–2019). Preliminary S. calcitrans in this zone was bimodal. Data These preliminary results could be focusing the fly control from the beginning of the Ana Laura Pèrez Sarasqueta1, Anderson spring to avoid the peak of autumn. Saravia3, Maria Ernestina Olhagaray Torres1, Cèsar Fernando Echaides Ortiz2, Eleonor Castro Janer1 PS02.33 Orientation of Stable Fly Larvae 1Departamento De Parasitologia Veterinaria, Facultad De Veterinaria, Universidad De La Dr. David Taylor1, Emily Kirkvold1, Dr. Kristina Republica, Uruguay., Montevideo, Uruguay, Friesen1, Dr. Dennis Berkebile1 2Intendencia de Montevideo, Montevideo, 1USDA-ARS-AMRU, Lincoln, United States Uruguay, 3Instituto Nacional de Investigaciòn Agropecuaria, Colonia, Uruguay Stable fly larvae can detect odorants within their developmental substrates and orient in Musca domestica and Stomoxys calcitrans response to them. Ammonium is the strongest are the main pests in confined animals and attractant eliciting a response from 75% of domestic farms with a worldwide distribution. larvae. Most of the esters examined acted as In Uruguay, the population dynamics of these attractants larvae as well. Alcohols, carboxylic flies are unknown, and this information is acids, ketones, aldehydes, aromatics and fundamental to their control. sulfides elicit little response. Stable fly larvae The study was conducted in a dairy farm in appear to use chemical cues different from the department of Colonia and in a wildlife those used by gravid females for identifying reserve, Parque Lecocq, located in the preferred developmental substrates. department of Montevideo, from February 2018 to January 2019. Dairy farm has 250 milking cows and the Zoo approximately 400 animals in 10 paddocks. Three models of traps were used to collect these flies: the Nzy, emergency traps and a homemade trap made with yellow plastic bottle with natural bait. The traps were placed in strategic sites, according to the behavior of each gender. In the Lecocq Park Zoo, 14,998 insects were captured. The abundance of M. domestica and S. calcitrans was very low. Only 70 specimens of M. domestica and 322 of S. calcitrans were

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 250 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.34 Distribution and Dynamics of PS02.35 Molecular Detection of Eimeria Canine Generalized Leishmaniosis in France: Species Among Chickens in KwaZulu-Natal Results of the 4th National Questionnaire Province, South Africa Based Survey with Veterinary Clinics Mr. Abiodun Fatoba1, Dr. Oliver Zishiri1, Prof. Patrick Bourdeau1, Laetitia Imparato1, Fanny Damer Blake3, Dr. Sunday Peters2, Dr. Jeffrey Alexandre1, Mailys Hilary1 Lebepe1, Prof. Samson Mukaratirwa1, Dr. 1Veterinary School of Nantes-oniris - France, Matthew Adeleke1 Nantes, France 1University Of Kwazulu-natal, Durban, South Africa, 2Berry College Mount Berry, Mount Canine Generalized Leishmaniosis Berry, United State, 3The Royal Veterinary (L. infantum) is widely distributed in College, Hawkshead lane, United Kingdom Mediterranean countries, including traditionally the South East part of France. Coccidiosis is a renowned intestinal disease Three previous National surveys (Bourdeau of livestock which impair growth and et al 1986-2004-2011) have shown a productivity. The impact of this disease has progressive extension of the endemic area. caused drastic production loss in poultry A fourth questionnaire-based survey with industry. Effective prevention and control of veterinary clinics was conducted in summer this disease hinges on proper identification. 2017 covering the period 2012-2017. For Despite the prevalent of this parasite species each of the 620 participating veterinary in different regions, there is little or no clinics the number of annual clinical cases, information of these species in South Africa. autochthonous (infection considered The objective of this study was to examine acquired within the range of the territory of the occurrence of Eimeria parasite using activity of the clinic) or not, were collected. molecular markers.To our knowledge, this is From the 90 administrative areas (= French first molecular survey of Eimeria parasite in departements), clinics with more than 10 KwaZulu-Natal province. In this study, 300 annual cases were found in 21 (20 in 2011). In fecal samples were collected from 12 different 33 areas, the average number of annual cases farms in KwaZulu-Natal province among per clinic was >1 (26 in 2011). Autochthonous which 102 (34%) were positive for Eimeria cases were suspected in 13 areas outside of infection. Four different species of Eimeria vis- the endemic zone, with kennel or familial a-vis: E. tenella, E. mitis, E. acervulina and E. foci in 6 and including 4 where the sand fly maxima were detected using species specific vector has never been captured and is not primers targeting ITS-1 regions with E. tenella supposed to be present. The endemic area being the most prevalent. Multiple infections covers 29 departements (26 in 2011) with with two or more species of Eimeria parasite kennel or familial foci detected in 22 of them. were also detected in some of the samples As observed for the period 2004-2011 and with E. tenella and E. maxima being the predicted, the northern progression of the most common combination. Single infection disease continued on the 2012-2017 interval. prevalent with E. tenella was also detected The estimated progression of surface of the in some of the samples examined. The endemic territory was 9%. In France the “wave report of this study shows that Coccidiosis effect” described in 2011 is maintained in the is widespread in chickens and a concerted periphery of endemic territories, whereas a strategy to prevent and control this disease is stabilisation or even a decrease of prevalence imperative in KwaZulu-Natal province. is suggested in the traditional center of the endemic zone. However, in the endemic zone, diagnosis was made on dogs originating from new places in 22 areas.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 251 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.36 Molecular Characterization of not be able to detect T. evansi DNA from Trypanosoma Evansi in Seventeen Isolates two in vitro cultivation isolates. There were from Five Natural Infected Hosts and In Vitro 3 among 17 isolates that lacked the Rotat 1.2 Cultivation in Thailand gene. Here we are the first T. evansi typing report in Thailand. Dr. Ketsarin Kamyingkird1, Miss Siriphon Tontong1, Dr Saruda Tiwananthakorn2, Dr. Ruttayaporn Ngasaman3, Dr. Burin PS02.37 3D-Holotomographic Analysis Nimsuphan1, Mr. Wissanuwat Chimnoi1, Miss of Respiration of Eimeria Bovis Oocyst Suchada Choocherd1, Dr. Marc Desquesnes1,4,5, Sporogony Dr. Nuttaphon Manojai6, Dr. Thiantada Bodhipongsathorn7, Dr. Tawin Inpankeaw1 Dr. Sara Lopez Osorio1, Dr. Zahady D 1Department Of Parasitology, Faculty Of Velasquez2, Dr. Ivan Conejeros2, Dr. Habil Anja Veterinary Medicine, Kasetsart University, Taubert2, Dr. Habil Carlos Hermosilla2 Bangkok, Thailand, 2Department of Veterinary 1Universidad De Antioquia - Justus Liebig Biosciences and Veterinary Public Health, University, Medellin, Colombia, 2Institute Faculty of Veterinary Medicine, Chiang Mai for Parasitology, Justus Liebig University, University, Mae Hiae, Thailand, 3Faculty Giessen, Germany of Veterinary Science, Prince of Songkhla University, Hatyai, Thailand, 4CIRAD, UMR Eimeria species are apicomplexan protozoan InterTryp, Bangkok, Thailand , 5InterTryp, parasites with high economic impact in Univ Montpellier, CIRAD, IRD, Montpellier, livestock. Oocysts of most enteropathogenic France, 6Hang Chat District Livestock Office, Eimeria spp. are shed by defecation into Lumpang, Thailand, 7Lumpang Small Animal environment as un-sporulated parasitic stages. Hospital, Lumpang, Thailand Exogenous oocyst development, namely from un-sporulated into sporulated infective status Trypanosoma evansi the causative agent for (i. e. sporogony) is only possible in aerobic trypanosomiasis in various vertebrate animals metabolic conditions. The sporogony process worldwide. There are two type of T. evansi can exclusively be observed in alive oocyts, as including type A and B. T. evansi type A was commonly used standard staining procedures predominantly niche and regularly reported do not penetrate highly resistant oocyst in West and East Africa, South America wall, thereby hampering permanent oocyst and China. T. evansi type B is restricted circumplasm labeling. Furthermore, it is difficult in Kenyan camel. T. evansi has endemic in to prepare Eimeria oocysts for transmission various kind of hosts with high mortality in electron microscope (TEM) by conventional horses, interfered animal health and reduced techniques explaining why TEM ultraestructural livestock production in Thailand. However, analyses have focused on endogenous there is no report that indicate the T. evansi merogony- and gamogony-stages, but never so type in Thailand. The aim of this study was to far on exogenous sporogony. In this study, we characterize T. evansi type using molecular document for the first time in vivo respiration technique. Seventeen DNA isolates of T. evansi of E. bovis oocyst sporogony using live cell infected blood collected from 1 dog, 1 buffalo, imaging microscopy techniques. For achieving 2 beef cows, 4 dairy cows, 7 horses and 2 sporogony, E. bovis oocysts (strain H) were re- from in vitro cultivation were extract and suspended in potassium bichromate K2Cr2O7 used for characterization. Specific primers solution (2%, w/v), at room temperature (RT) at including Rotat 1.2 (type A specific), EVA B 25 °C with constant oxygenation. Daily, oocysts (type B specific), ITS 2, VSG, Mini (minicircle were documented using 3-D holotomographic kDNA) and NC (ITS 5.8-2 region) were used. microscopy (3D Cell Explorer®, Nanolive) Phylogenetic analysis was conducted. Results to explore instantly live sporogony in 3D showed that there were 14/17 isolates of T. dimensions without any labeling or chemical evansi type A and 0/17 of T. evansi type B. marker staining. 3D-E. bovis oocyst images NC primer targeting ITS 5.8-2 region was very were digital stained based on the cell’s physical sensitive and be able to detect T. evansi DNA refractive index using STEVE® software in all 17 isolates. VSG and ITS 2 primers were (Nanolive).

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 252 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.38 Seroprevalence of Toxoplasma PS02.39 Validation of Single-Tube Nested Gondii Infections in Dogs and Cats of Korea, Real-Time PCR and Genetic Sequencing 2017–2018 for Detection and Characterization of Cryptosporidium Spp. in Birds Jinhyeong Noh1, Mi-Sun Yoo1, Yeojin Park1, Jiyeon Lim1, Subin Min1, Bang-Hun Hyun1, Yun Dr. Bruna Santana1, Ms. Isabela Nascimento1, Sang Cho1 Dr. Marcelo Meireles1 1Parasitic and Insect disease laboratory, 1Universidade Estadual Paulista (unesp), Animal and Plant Quarantine Agency, Campus De Araçatuba, Faculdade De Gimcheon-si, Republic of Korea Medicina Veterinária., Araçatuba, Brazil

Cats are the natural reservoir of Toxoplasma Molecular detection and characterization of gondii and excrete the resistant oocyst to Cryptosporidium spp. in fecal samples are usually environments, and dogs play a role in the performed by two steps conventional PCR assays mechanical transmission of the protozoa. followed by agar gel electrophoresis and genetic The objectives of this study were to assess sequencing. The objective of this study is to the prevalence and risk factors of T. gondii validate a protocol of single-tube nested real-time infection in dogs and cats, which have an PCR assay followed by melting curve analysis and important role in pulic health, by serological genetic sequencing to detect and characterize diagnostic test. the species and genotypes of Cryptosporidium in birds. The assay was performed to amplify Serum samples were collected from a fragment of the 18S rRNA gene using DNA companion and shelter animals in veterinary samples extracted from 249 fecal samples from clinic and in animal shelter. The presence birds (47 and 202 tested positive and negative, of T. gondii antibodies was analyzed using respectively, by nested PCR targeting the 18S the commercial kit based on enzyme-linked rRNA gene), two sets of primers with different immunosorbent assay. annealing temperatures, and the SsoFast EvaGreen Supermix (Bio-Rad) in the CFX96 A total 2,169 blood samples of dogs and cats real-time PCR system (Bio-Rad). Conditions of from veterinary clinic and animal shelter. T. the assay were: 98 ºC for 2 min was first followed gondii antibodies were detected in 45(2.1%) by 20 cycles of denaturation at 98 ºC for 5 s and of the 1,215 positives in cats. The positive annealing/extension at 70 ºC for 30s. This was rates of shelters(17.0%) was higher than followed by 35 cycles of denaturation at 98ºC companions(3.3%) in cats. On the other for 5 s, annealing at 63 ºC for 5 s, and extension hands, T. gondii antibodies were examined at 72º C for 30 s. Fluorescence signal acquisition as positive in 17(1.1%) of the 1,613 dog occurred at the second cycling step. Melting curve blood samples and shelter and companion analysis was performed from 70 °C to 95 °C. dogs had 0.7% and 0.1%, respectively. The Amplified fragments were purified using ExoSAP- seroprevalence of T. gondii according to IT (Thermo Fisher Scientific) and submitted to district is shown as follows; Gyeonggi(1.2%), bidirectional sequencing for confirmation of Jeolla(0.2%), Jeju(0.2%), Gyeongsang(0.1%) Cryptosporidium species/genotype. Melting and Chungnam(0.1%). temperatures of the bird Cryptosporidium species and genotypes were 79.6°C, 79.6°C, 79.4°C, Serological surveillance of toxoplasmosis 80°C, and 80.2°C for Cryptosporidium baileyi, in dogs and cats was investigated in Cryptosporidium meleagridis, avian genotype this study. Differences are shown in I, Cryptosporidium galli and avian genotype III, seroprevalence between dogs and cats, respectively. The frequency of samples positive shelters and companions by region. Further for Cryptosporidium spp. by nested real-time PCR epidemiological study is required to was 80/249 (32.1%). Our results show that single- dertermine sources of T. gondii infection in tube nested real-time PCR could be used as an dogs because information on the infectious alternative to conventional nested PCR with the status of T. gondii in dogs is important for advantages of lower turnaround time and lower assessing the risk to public health. risk of carry-over contamination.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 253 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.40 Efficacy of a Single Oral Universidade Estadual “Júlio de Mesquita Administration of Afoxolaner Alone or in Filho” – UNESP, Sorocaba, Brazil, 3Embrapa Combination with Milbemycin Oxime Against Pecuária Sudeste – CPPSE, São Carlos, Brazil Ixodes Hexagonus in Dogs The aim of this study was to evaluate the Prof. Frederic Beugnet1, Dr. Wilfried Lebon1, larvicidal potential of formulations developed Dr. Pascal Dumont1, Dr. Marielle Servonnet1 from a nanocarrier system based on 1Boehringer-ingelheim, Lyon, France cypermethrin (CPM) + chlorpyrifos (CPF) and its association with bioactives isolated from The efficacy of afoxolaner (NexGard® and plants. The CPM + CPF based formulations NexGard Spectra®, Boehringer-Ingelheim), were developed from Nanostructured administered once orally at the minimum Lipid Carriers (NLC) and associated with recommended dose, was assessed in substances isolated from plants commercially dogs experimentally infested with Ixodes acquired (Sigma-Aldrich): cytral (Form. A), hexagonus ticks. menthol (Form. B), and limonene (Form. C). The formulations A, B, C and NLC (alone) The study was a blinded, negative controlled were then evaluated against R. microplus clinical efficacy study using a randomized block larvae from 100 to 0.78 l.mL-1 concentrations design. Twenty-four purpose bred Beagle dogs, by the Larval Packet Test (LPT), in triplicates, 12 females and 12 males were included. Dogs negative (water) and positiveµ (Colosso® were randomly allocated either to the negative CPM+CPF at 1.25 L.mL-1) control. After 24h control group, or to one of the two treated of incubation (± 27 °C; > 80% RH) the count groups. Infestations were performed with 50 of live and dead larvaeµ was performed. The adult I. hexagonus ticks on days -2, 7 and 28. On results were analyzed by ANOVA One-way day 0, dogs in groups 2 and 3 were treated with followed by Tukey’s test. Formulations A, NexGard ® (afoxolaner) or NexGard Spectra® B and C caused 100% mortality at 1.56, 3.12 (afoxolaner + milbemycin oxime), respectively. and 6.25 L.mL-1, respectively, with a dose- Live tick counts were conducted 48 hours after dependent effect, and significant differences treatment (day 2) and 48 hours after each (p≥0.05).µ The positive control had 100% subsequent infestation (days 9 and 30). efficacy, and the formulations A and B caused 81.31 and 76.27% mortality at 0.78 L.mL-1. In both treated groups, afoxolaner was Moreover, CLN caused mortality > 40% at 100% effective against existing infestations 50 L.mL-1. It was demonstrated thatµ the (p<0.0001). Regarding the re-infestations, nanocarrier system evaluated was effective, overall efficacy of afoxolaner was 100% at day 9 sinceµ the active compouds, even reduced, and 98.5% at day 30. caused mortality rates similar to those of the commercial reference product. In 100 NexGard and NexGard Spectra chewable mL, Colosso® and Formulations (A, B and C) tablets administered once orally at the contained, respectively: 15.0g and 0.1875g minimum recommended dose were highly of CPM; 25.0g and 0.3125g of CPF; 1.0g of effective against I. hexagonus infestations for citronellal and 0.0125g of isolated from plant the 4 weeks duration of the study. (cytral, menthol or limonene) in addition to 5.87*1015 of CLN in the formulations of present study. Thus, the CLN system can PS02.41 Nanocarrier Formulations Against be considered a possible option on the Rhipicephalus (Boophilus) Microplus Larvae development of new acaricides. New studies are necessary to elucidate the function of Dr. Amanda Figueiredo1,3, Dr. D.F. Cola2, Dr. these associations, and to validate its efficacy Y.A. Gainza1,3, Dr. L.F. Fraceto2, Dr. A.C.S. against R. microplus. Chagas3 1Departamento de Medicina Veterinária Preventiva, Universidade Estadual “Júlio de Mesquita Filho” – UNESP, Jaboticabal, Brazil, 2Departamento de Engenharia Ambiental,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 254 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.42 Diversity and Geographic PS02.43 Candidatus Neoehrlichia Mikurensis Distribution of Brown Dog Ticks in the in Ixodes Ricinus Ticks in the Czech Republic United States Dr. Jaroslav Ondruš1,2, Prof. Adam Novobilský3, Michelle Lentile1, Kellee Sundstrom, Meriam Dr. Pavel Kulich3, Prof. Pavel Široký1,2 Saleh, Susan Little 1University Of Veterinary and Pharmaceutical 1Oklahoma State University, Stillwater, United Sciences Brno, Brno, Czech Republic, 2CEITEC States Veterinary and Pharmaceutical University, Brno, Czech Republic, 3Veterinary Research Brown dog ticks (Rhipicephalus sanguineus Insititute, Brno, Czech Republic sensu lato) are now known to exist as a species complex comprised of several distinct Candidatus Neoehrlichia mikurensis (CNM) lineages. Although surveys are limited, two is a tick-borne emerging intracellular lineages have been described in the United bacterium recently found in ticks from over States to date: temperate and tropical. To 20 Eurasian countries. Small rodents serve more fully characterize the distribution of R. as a reservoirs. Despite this bacterium is sanguineus s.l. throughout the United States, acknowledged as a human pathogen since a 12S rRNA gene fragment was amplified and 2010, only little is known about its life cycle, sequenced from brown dog ticks removed transmission, structure or target cells. In this from 153 dogs and 3 cats from different study, distribution and prevalence of CNM in locations across 14 of the 50 states from Ixodes ricinus using PCR was investigated in March 2018 through January 2019; a nested the Czech Republic. In addition, transmission PCR assay targeting an Ehrlichia spp. 16S electron microscopy (TEM) was used to rRNA gene fragment was also used to assess identify and visualize CNM in ticks. prevalence of infection with Ehrlichia canis in ticks. Brown dog ticks were submitted in To identify the possible hotspots of every month of the year except February. “neoehrlichiosis”, over 13 600 ticks were A majority (79.5%; 124/156) of brown dog collected by flagging in surroundings of tick populations evaluated were temperate major Czech cities. Ticks were subsequently lineage, while only 32/156 (20.5%) were classified by stage and sex and pooled tropical lineage. As expected, most (26/32; into groups of 5 individuals. In total, DNA 81.3%) tropical lineage R. sanguineus s.l. was isolated from 2 666 groups. CNM were from areas with an annual average daily detection was done by conventional PCR. temperature > 20°C (68°F), including Arizona, Ca. Neoehrlichia mikurensis was detected Florida, Hawaii, Nevada, and southern Texas. in I. ricinus in 150 of 169 localities examined. However, several (n=6) tropical R. sanguineus Furthermore, several sites were identified as s.l. infestations were found on dogs from hotspots for neoehrlichiosis, mainly in areas unexpected geographic locations in California, with high tourism activity. Kentucky, Michigan, Minnesota, Oklahoma, and northern Texas; follow up conversations The ultrastructure of CNM in the vector host with veterinarians and owners in these cases was examined on the ultrathin sections of suggested some of these infestations resulted infected tissues of I. ricinus using TEM. Ticks from recent travel of dogs from tropical areas. flagged in the locality of high CNM prevalence Ehrlichia canis infection was not detected in were allowed to feed in the artificial in vitro any of the R. sanguineus s.l. ticks in this study. feeding system. After dissection, individual These data confirm that most brown dog ticks organs were prepared for TEM analysis. Since in the United States are temperate lineage and this pathogen has not yet been cultivated suggests that pet travel occasionally results in and the surface structures remain unknown, translocation of tropical lineage R. sanguineus the screening was somewhat hindered by s.l. to new areas. unavailability of labeled probes. However, structures resembling the CNM bacteria in mammalian tissues were found. In summary, we detected Ca. Neoehrlichia mikurensis in the close proximity of the majority of sizable

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 255 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Czech cities and described putative Ca. was found between the molecular structure Neoehrlichia mikurensis individual bacteria of the compounds and their activity. Cyclic within the salivary glands of the European tick compounds which containing ten carbon I. ricinus. molecules in their chemical structure were more toxic to larvae compared to acyclic This study was supported by grants IGA ones. It is concluded that the methodology of 205/2018/FVHE, IGA 201/2019/FVHE and evaluation by means of larval mortality time AZV 16-33934A. was able to determine which essential oil had the highest toxicity in vitro tests.

PS02.44 Mortality Time of Rhipicephalus Microplus Larvae Immersed in Essential Oils PS02.45 Mortality Time of Rhipicephalus Microplus Larvae Immersed in Different Leandro Rodrigues1, Dr. Cecília José Solvents Veríssimo1, Dr. Luciana Morita Katiki1, Karen Tiemi Akashi3, Dra. Mariana Santos De Leandro Rodrigues1, Dr. Cecília José Miranda1, Ms. Poliana Araújo Pacheco2, Dr. Veríssimo1, Karen Tiemi Akashi3, Dr. Luciana Rodrigo Giglioti1 Morita Katiki1, Dr. Mariana Santos De Miranda1, 1Institute of Animal Science, Nova Odessa, Poliana Araújo Pacheco2, Dr. Rodrigo Giglioti1 Brazil, 2University of São Paulo, Piracicaba, 1Institute of Animal Science, Nova Odessa, Brazil, 3State University of Maringá, Maringá, Brazil, 2University of São Paulo, Piracicaba, Brazil Brazil, 3State University of Maringa, Umuarama, Brazil Rhipicephalus microplus ticks are responsible for damage to the health of Researchers are currently looking to study its hosts and generate economic losses to and develop new acaricidal products to the livestock sector. The development of control Rhipicephalus microplus tick. The parasitic resistance to most commonly used objective of this work was to evaluate commercial acaricides has led to the search the mortality time of R. microplus larvae for alternative control. In this scenario, the immersed in liposoluble solvents (glycerol, present study aimed to evaluate the toxicity vaseline, castor oil, xylol) and water soluble of essential oils by the mortality time of R. solvents (acetic acid, acetone, Tween 80, microplus larvae in direct contact with oils. DMSO, propylene glycol, ethanol, methanol Among 20 terpenes studied, 14 were cyclic and isopropyl alcohol) in vitro. The solvents (carvacrol, thymol, 1,8-cineole, anethole, were tested pure (100% concentration) and cinnamaldehyde, L-limonene, vanillin, carvone, were diluted in water - negative controls eugenol, menthol, terpinolene, -pinene, (A) or soybean oil (OS) according to their D-limonene and - terpinene) and six were solubility to perform 50, 25, 12.5, 6.25, 3.125 acyclic chains (citral, citronellal,β citronellol, and 1.56% concentrations. The test consisted geraniol, linalool αand nerolidol). Mortality of immersing approximately 100 larvae time was compared with negative controls of R. microplus in 200 l of the solvent (soybean oil) and two positive controls tested in a 48 well plate. All larvae were (commercial Pour on). The test consisted in observed with a magnifyingμ glass and with immersing approximately 100 R. microplus a cronometer, we verified the time in which larvae in 100 l of each oil in 100% dose in 100% of the larvae were dead. All solvents, 48 well polyethylene plates and verifies the concentrations and the control treatments time when 100%µ of larvae were dead, with the were tested in triplicates. Data were analyzed aid of a timer. The ten oils with the shortest by PROC GLM procedure of SAS, whose mortality time were diluted in soybean oil model included the fixed dose effect. Each to form the doses of 50, 25, 12.5, 6.25, 3.125 solvent was analyzed individually. The means and 1.56%. The essential oil with higher larval between the different doses were compared toxicity was thymol, in which the lowest dose by the Tukey’s test. All solvents and their (1.56%) had the shortest mortality time (15 concentrations differed from controls (A) min) compared to the others. A relationship and (OS) (p 0.05) which had a mortality

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WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 256 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

time of 4320 and 2400 minutes respectively. PS02.47 Efficacy of Different Concentrations It was observed that larval mortality time is of Duddingtonia Flagrans Chlamydospores dose dependent for most solvents. Among against Different Levels of Faecal Egg the liposoluble solvents, xylol presented the Counts of Cattle Gastrointestinal Nematodes shortest mortality time of larvae ranging from 0.2 min (100% dose) to 185 min (1.56% dose). Sara Zegbi1, Dr. Federica Sagües1, Dr. Carlos Among the water soluble solvents, acetic acid Saumell1, Prof. Lucía Iglesias1, Inés Guerrero1, had the shortest mortality time (p 0.05) with Dr. Silvina Fernández1 variation from 0.6 min (100% dose) to 97 min 1Área de Parasitología y Enfermedades (1.56% dose). We concluded that the˂ solvents Parasitarias, Centro de Investigación used in acaricidal products can influence the Veterinaria de Tandil (CIVETAN) (UNCPBA- mortality of larvae. CICPBA-CONICET), Facultad de Ciencias Veterinarias, UNCPBA, Tandil, Argentina

PS02.46 Exotic and Disease Vectoring Ticks The nematophagous fungus Duddingtonia on White-Tailed Deer in Southern Texas flagrans is a biocontrol agent that reduces the number of infective larvae of gastrointestinal Dr. Donald Thomas1, Dr. Roberta Duhaime2 nematodes in animal faeces, thus, lowering 1USDA-ARS, Edinburg, United States, 2USDA- pasture infectivity. However, the minimum APHIS, Laredo, United States amount of fungal chlamydospores in faeces required for this effect is largely unknown. White-tailed deer were scratch inspected The following in vitro study aimed to for ticks during the 2018 hunting season determine the fungal efficacy of four different at the Laguna Atascosa National Wildlife chlamydospores concentrations against Refuge in Cameron county TX. Only four three different levels of cattle faecal egg of the 73 deer examined were negative for counts. The chlamydospores concentrations infestation by ticks. A total of 3380 ticks tested were 11000, 6250, 3000 and 1000 were counted and identified to one of seven chl/g faeces. Faeces from naturally-infected species. Five of the encountered tick species calves were used to obtain faecal egg are precinctive to the neotropics and just counts of 100, 480 and 840 epg. Ten faecal enter the United States in the border area of cultures of 10 g faeces each were set up Texas: Amblyomma inornatum, A. mixtum, for each chlamydospores concentration/ A. tenellum, Anocentor nitens and Boophilus faecal egg count combination, plus one microplus. The latter two species, the tropical control group for each faecal egg count horse tick and the southern Cattle Fever Tick, level without chlamydospores. All cultures both one-host ticks, are important economic were kept at room temperature (20-26ºC) pests of livestock and thus, it is of some for two weeks and then L3 were recovered concern that they use white-tailed deer as a from each individual culture by overnight sylvatic reservoir. Notably, although deer have baermannisation, counted and identified. The been reported as an incidental host, the very larval reductions by D. flagrans, in decreasing high numbers (2922) including larvae and order of chlamydospores concentrations, nymphs as well as adults, suggests that deer were: 100% (P<0.0001), 99% (P<0.0001), are the native host of the “Horse tick” given 92% (P=0.0032) and 77% (ns) compared that horses are not native to the western to the 100 epg control; 100% (P<0.0001), hemisphere and A. nitens does not occur in 98% (P<0.0001), 98% (P=0.0002) and 92% the Old World. The gulf coast tick, another (ns) compared to the 480 epg control; and pest of livestock was common and coexistent 100% (P<0.0001), 100% (P<0.0001), 99% with the other Amblyomma spp. in this deer (P<0.0001) and 96% (ns) compared to the herd. Finally, the detection of 21 specimens of 840 epg control. There were no differences Ixodes scapularis, the vector of Lyme disease, between the three levels of faecal egg counts suggests that this species may not be as at any given fungal concentration, which rare in south Texas as previous surveys have indicates that the numbers of nematode eggs indicated. in cattle faeces would not be a determining factor for the efficacy of D. flagrans. These

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 257 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

results suggest that the concentration of of egg hatching and mortality of larvae at 1000 chl/g faeces would not be sufficient to concentration ranges of 0.78 - 12.5 mg/ml. achieve a significant fungal efficacy; however, The result obtained from this study suggests it could be argued that, in practice, larval that H. pachyanta possess anthelmintic reductions of 77% to 96% would contribute to effects against H. Contortus and validates the control gastrointestinal nematodes. folkloric use of the plant in the management of H. Contortus infections.

PS02.48 In Vitro Anthelmintic Activity of Hymenodictyon pachyanta Stem Bark Extract PS02.49 Humoral Response of Canaria Sheep and Fractions Against Haemonchus contortus Vaccinated With a Recombinant Teladorsagia Circumcincta Prototype Vaccine Prof. Olorunfemi Jegede1, Dr. Olutayo Olayemi1, Dr. Michael Onakpa1 Cynthia Machin1, Julia N. Hernández1, Tara 1University Of Abuja, Abuja, Nigeria Pérez-Hernández1, Yolanda Corripio-Miyar2, Harry Wright2, Dan R.G. Price2, Tom N. The developoment of helminthe resistance McNeilly2, Alasdair J. Nisbet2, Jacqueline B and high cost of synthetic anthelmintics has Matthews2, Jorge F. González1 prompted the need for an in vitro anthelmintic 1Universidad De Las Palmas De Gran Canaria, evaluation of crude stem bark extract and Arucas, Spain, 2Moredun Research Institute, fractions of Hymenodictyon pachyanta United Kingdom plant as an alternative in the management of endoparasites including Haemonchus Among the potential alternatives to contortus which is one of the most prevalent anthelmintics against gastrointestinal parasitic nematodes in small ruminant farming nematodes, vaccines are an environmentally globally. Hymenodictyon pachyanta stem bark friendly and promising option. An efficacious is used as anthelmintic by indigenous farmers recombinant sub-unit vaccine against in Nsukka, Enugu State and Gwagwalada, Teladorsagia circumcincta has been tested Federal Capital Territory, Nigeria. The stem in Texel-cross sheep (Nisbet et al., 2013; bark of H. pachyanta were collected from 2016; 2019). Here, we evaluated the effect the field in Nsukka, Enugu State, air dried, of this prototype vaccine in another sheep pulverized and extracted with 80% Methanol. breed, the Canaria Sheep (CS). In vaccinates The extract and fractions of H. pachyanta compared to adjuvant-only control sheep, were tested on the egg hatch inhibition assay female worms were found to be significantly (EHIA) and the larval development inhibitions shorter and had fewer eggs in utero. Although assay (LDIA) and compared with Albendazole mean worm burden and faecal egg counts the positive control. The concentrations for were lower in vaccine recipients, the levels the plant extract, fractions and Albendazole were not significantly lower that found in the used for the study were 0.78, 1.56, 3.125, 6.25 controls. Significant negative associations and 12.5mg/ml. The results showed that the between antigen-specific IgG and IgA and crude extracts, fractions and Albendazole at parasitological parameters (worm burden/ concentration dose of 12.5 mg/ml produced worm fecundity) were found. These data 100% inhibition of egg hatching and larval add to our understanding for the further development of Haemonchus contortus. development of this vaccine for sheep. Although, there was no significant difference Acknowledgements: European Union’s (p>0.05) in the mean percentage egg hatch Horizon 2020 Research and Innovation and larval development inhibition of the programme under the Grant Agreement No. crude extract and fractions when compared 635408 (PARAGONE). with Albendazole. However, significant difference (p<0.05) was observed with n-butanol fraction which inhibited 96.17% of egg hatchability. The extract, fractions and Albendazole showed ovicidal and larvicidal activity and produced over 50% inhibition

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 258 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.50 A Predation Test to Evaluate the lower in the test cultures (p=0.048), but it Efficacy of Macrochelidae Mites as Biological was unfeasible to recover mites from the Control Agents for Parasitic Nematode substrate. The procedure was repeated with Larvae horse rectal sample used to prepare six repetitions with three coprocultures each. Two Karina Araújo dos Anjos1,5, Shirley Batista da cultures from each repetition received a single Silva Araújo1,5, Oswaldo José Christe Adorno2, female Holostapella and the third served as Tatyana Sacchi Carmona Rodrigueiro3, control. The additional cultures with acari Leonardo Costa Fiorini4, Fernanda Calvo were placed in the Berlese-Tullgren apparatus. Duarte4, Paulo Henrique Selbmann Sampaio4, Five cultures yielded live mites, and four Márcia Cristina Mendes4 produced two. Nematode larvae counts were 1Master’s Program in Health, Food and significantly lower in test cultures (p=0,037). Environmental Safety in Agribusiness, The current model has yet to be refined, but Biological Institute, São Paulo, Brazil, it offers positive evidence concerning the 2Coordination of Integral Technical Assistance potential use of these predators as biological (CATI), Guaratinguetá, Brazil, 3Technical State control agents for parasitic nematode larvae School Adhemar B. Heméritas, Paula Souza on pastures. Center, São Paulo, Brazil, 4Animal Health Research Center, Biological Institute, São Paulo, Brazil, 5Coordination of Superior Level PS02.51 Evaluation of the Fecal Dx® Antigen Staff Improvement (CAPES) scholarship, Test for the Diagnosis of Nematodes with Finance Code 001, Brasília, Brazil Carnivorous Samples from a Germany Zoo

Macrochelidae acari are foretic predators Dr. Jinming Geng1, Dr. David Elsemore1, found in soil, decomposing organic matter Manuela Enzelberger2, Katrin Baumgartner2, and feces. A survey conducted in dairy farms Simone Pürkel2, Hermann Will2, Majda at the municipality of Guaratinguetá - SP, Globokar3, Alaa Alnassan3, Nikola Pantchev3 Brazil recovered Allogynaspis flechtmanni, 1IDEXX Laboratories, Westbrook, United Glyptholaspis americana, Glyptholaspis States, 2Tiergarten Nürnberg (The Nuremberg saprophila, Holostapella bifoliata, Macrocheles Zoo), Nuremberg, Germany, 3IDEXX Vet Med insignitus, Macrocheles mammifer, Labor GmbH, Ludwigsburg, Germany Macrocheles merdarius, Macrocheles muscaedomesticae, Macrocheles robustulus, Common nematodes in zoo felids and Macrocheles glaber, Macrocheles scutatus canids are Toxocara sp., Toxascaris leonina, and Macrocheles subbadius from partially dry Ancylostoma sp., Uncinaria stenocephala cattle dung patches. Mites were fed Musca and Trichuris vulpis. Apart from various domestic eggs and infective nematode larvae clinical signs of gastrointestinal parasitosis from coprocultures of cattle, horses and like anorexia, weight loss, diarrhea or bowel sheep. A colony of Holostapella bifoliata was obstruction (especially in young animals), T. established and assigned to a predation test canis and T. cati are also important zoonoses. on coprocultures. Feces collected directly The diagnosis of these diseases is usually from the rectum of seven cattle were blocked dependent on fecal egg flotation, which according to their respective EPG counts. is time-consuming and not user-friendly. Each sample was halved and each half sample Compared with fecal flotation, the IDEXX Fecal was mixed with vermiculite for coproculture. Dx® antigen test (Fecal Dx) is an ELISA that One was assigned to the test group and its provides detection of infection in the absence pair served as control. After two days at 30°C of an egg observation. However, few studies ± 2°C e 70% ± 10% humidity, five female and have been reported about the application of two male mites were placed in each test Fecal Dx for diagnosis of ascarids, hookworms culture. After eight days, infective nematode and whipworms in wild carnivores (cheetah, larvae were recovered and counted. Mann- lynx, leopard, tiger, lion, wolf, polar bear, Whitney test was used to compare recovery red panda, meerkat, fishing cat and yellow- rates calculated on the base of the initial throated marten). Since Fecal Dx was designed EPG counts. Larvae counts were significantly to detect the secreted/excreted proteins

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 259 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

from Ancylostoma caninum, Toxocara canis, evaluations 3, 4 and 5, and EPG counts Toxocara cati and Trichuris vulpis in domestic superior to 1000 for McMaster and Mini- canines and felines, very little is known about FLOTAC) and Untreated. Prior to the trial’s its performance for the diagnosis of Ascaris, beginning, coprocultures were carried out hookworms and whipworms in the wild animals for the generic identification of helminths. mentioned above. There were 1142 comparable evaluations, and Haemonchus (67%) was the most prevalent In this study, Fecal Dx, with passive fecal genre in coprocultures. The FAMACHA© egg flotation as a reference method, was method showed 82.66% of the evaluations evaluated with 51 fecal samples from 11 in degrees 1 and 2. 69.44% and 67.51% of all different carnivores from a German Zoo. For evaluations presented EPG counts between 0 ascarids, 25.4% (13/51) of the samples were and 990, for the McMaster and Mini-FLOTAC positive by egg flotation, compared with methods, respectively. The FAMACHA© 39.2% (20/51) of the samples with Fecal Dx. method presented small correlation with EPG Cheetahs and leopards were most likely to counts, demonstrating that, the higher the be infected with ascarids compared to other egg mean counts, the more marked were the carnivores tested. In addition, cheetahs were degrees of anemia. Comparison of McMaster the only species in which hookworm and and Mini-FLOTAC techniques revealed that whipworm infections were found. Contrasting 86.25% of evaluations presented differences to flotation where no hookworm or whipworm from 0 to 490, with 133 results being identical, eggs were observed, Fecal Dx showed 5.8% 578 obtaining differences from 10-190 and (3/51) and 1.9% (1/51) of the cheetahs tested 274 from 200-490. Treated and Untreated were hookworm and whipworm positive, evaluations for each technique resulted in a respectively. 50% reduction in treatment frequency when FAMACHA© was applied (P < 0.05). Based on obtained results, it I possible to conclude PS02.52 Comparative Efficiency of the that all three methods presented expressive McMaster, Mini-FLOTAC and FAMACHA© helminths diagnosis results, with little Methods for Diagnosing Helminths in Sheep difference between them. However, when data were transformed into Treated and Untreated Dr. Willian Maciel1, Dr. Isabella Santos1, P Dr. animals, the FAMACHA© method stood out, Breno Cruz1, Dr. Carolina Buzzulini1, Mr. Daniel significantly reducing the needed frequency Melo1, Mr. Dalmo Quilis1, Dr. Lucas Gomes1, of treatments. Mr. Davi Salvador1, Ms. Ana Flávia Mendes3, Dr. Gustavo Felippelli1, Dr. Welber Lopes2, Dr. Alvimar Costa1, Dr. Gilson Oliveira1 PS02.53 Association Between Dirofilaria 1Universidade Estadual Paulista – UNESP, Immittis and Euocleus Aerophilus in Naturally Jaboticabal, Brazil, 2Universidade Federal Infected Dogs, in the Northern Outskirts of de Goiás – UFG, Goiânia, Brazil, 3Centro Buenos Aires and the Tigre Region Universitário Barão de Mauá - CBM, Ribeirão Preto, Brazil Dr. Gabriela Perez Tort1,2, Dr. Pablo Borrás3, Dr Jordana Gueijman1 The present study aimed to evaluate and 1Veterinary Virreyes Hospital, San Fernando, compare three methods of diagnosing ovine Argentina, 2Faculty of Veterinary Sciences helminths. For such, 48 matriarchs were Buenos Aires University, Buenos Aires, selected, with EPG (eggs per gram of feces) Argentina, 3Servicio de enfermedades counts superior to 400 eggs on a preliminary parasitarias e infecciosas Veterinaria Panda, McMaster screening. Fecal collections and Buenos Aires, Argentina evaluations of ocular conjunctiva were performed in all ewes, for evaluating parasite The aim of this report is to point out the burdens by the FAMACHA©, Mini-FLOTAC presence of both Dirofilaria immitis and and McMaster methods, every 14 days over Eucoleus aerophilus in dogs naturally an entire year. Results of all three techniques infected in the littoral region and the northern were dichotomized in Treated (FAMACHA© outskirts of Buenos Aires. Both parasites

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 260 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

can produce chronic cough or dogs can The authors present the first report in be asymptomatic. Eucoleus aerophilus, Argentina of Demodex gatoi in one cat. live attached to mucous membranes of Demodex gatoi is a short mite located in trachea and bronchi. The eggs (bipolar and the skin stratum corneum. Unlike D.cati, it asymmetric) can be found in faeces, measure causes itching and a contagious condition 60-70u x 35-40u, and contain a single cell in immunocompetent cats. The main when passed. Pets are infected by ingestion clinical signs are associated with moderate of the egg with an infective larva; eggs to intense pruritus. Diagnosis was based require a period of development in the soil on skin scraping, acetate tape, and fecal to become infectious. Dogs presented to The flotation copro-parasitological test to detect Veterinary Virreyes Hospital from 1997 to parasites. Identification is based on the typical 2018 were included in this paper. Of 35 dogs morphology of this Demodex. positive to Eucoleus aerophila (diagnosed by fecal flotation or endoscopy), 20 were It was a Feline, neutered female, 4 months positive to Dirofilaria immitis, (diagnosed by old, of Bengali breed referred to the clinic, Antigen test and Knott test) and 15 negative. with diarrhea and intense itching that started Of 410 positive to Dirofilaria immitis 20 were since her arrival at home. The cat was from positive to Eucoleus aerophilus. A complete a cattery in the city of Buenos Aires. She blood count, creatinine, ALT, urinalysis, showed pruritus, which manifested with urine culture, coproparasitological exam, scratching, excessive licking and bites all over thoracic radiography, electrocardiography her body. Inflammation of the bowel and were performed to all of them. The medical erythema of the head and neck were detected treatment sequence recommended by the at clinical examination. The blood count and American Heartworm Society was followed. blood chemistry showed no changes. FIV / Complications of heartworm disease or FeLV negative. Fecal studies were performed other diseases were managed before with fresh samples: modified Ziehl-Neelsen adulticide therapy was begun. Eucoleus stain, direct exam with physiological solution, was treated with fenbendazole 50mg/kg and Bembrock technique. Tritrichomona for 3 days before Heartworm treatment was foetus, Giardia lamblia, Cryptosporidium spp. begun. The adulticide treatment was made and Demodex gatoi were found. Trichography with melarsomine dihydrocloride, 2,5mg/ samples with acetate strips from affected and kg, all dogs were kept at the hospital for unaffected areas of the skin were taken as a fortnight. From 2002 on, all dogs were well as skin scrapings. Eggs and adult mites of treated with the alternate dose regimen. Demodex gatoi were observed. An analgesic protocol was started from one hour previous to the melarsomine injection. The cat was treated specifically against From 2009 on, patients have also received Demodex gatoi with Advocate spot on (Bayer doxycycline orally 10 mg /kg once a day to laboratory, Imidacloprid 100 mg, Moxidectin control Wolbachia. It is concluded that in the 10mg) repeated at day 15; negative results presence of either Dirofilaria or Eucoleus the were obtained at the end of the treatment. other should be suspected, in this region. Demodicosis is a rare parasitic disease in feline consulting room; this being the first report of Demodex gatoi in Argentina. PS02.54 First Clinical Case of Demodex Interestingly, diagnosis was made using a Gatoi in a Cat in Argentina fecal flotation. This disease should be then taken into account when facing a pruritic Gabriela Perez Tort1, Dr. Javier Cespedes2, Dr. cat mainly if the patient comes from feline Pablo Borras3 communities. 1Virreyes Veterinary Hospital, Buenos Aires, Argentina, 2Dr. Javier Céspedes Clínica Veterinaria, Rosario, Argentina, 3Servicio de Enfermedades Infecciosas y Parasitarias, Veterinaria Panda, Buenos Aires, Argentina

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 261 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.55 Use of a PCR to Identify the Main L3 from mixed GIN samples. The study will Gastrointestinal Nematodes Resistant to continue to improve the agreement between Anthelmintics in Cattle Farms, in Uruguay both morphological and molecular tests for our working conditions. Celeste Serrano1 1Instituto Nacional de Investigacion Agropecuaria, Tacuarembo, Uruguay PS02.56 Easy, Cheap and Convenient Method for the Detection of Different Species of The objective of this work was to set up Babesia molecular tests to identify the main resistant gastrointestinal nematodes (GIN) identified as Mr. Zohaib Shahid1, Dr. Muhammad Arfan resistant by coprocultures when performing Zaman1, Ms. Uzma Mehreen1, Ms. Warda the fecal egg count reduction test (FECRT). Qamar1, Professor Muhammad Fiaz Qamar1, Third stage larvae (L3) were obtained from Mr. Kazim Ali1 the FECRT conducted on 10 extensive cattle 1College of Veterianry and Animal Sciences, farms in the North of Uruguay during 2018. Jhang, Pakistan Morphological and molecular identification was done using L3 obtained for drug groups: Babesia is a disease transmitted by ticks. Ivermectin 1% (IVM), Levamisole (LEV), it is present in whole world. It has impact Ricobendazole (RBZ), Fenbendazole (BZ) on animal health as it causes severe anemia and untreated control. A commercial kit which leads to loss of meat and milk and NucleoSpin® Soil DNA (Macherey-Nagel) was has direct impact on human as babesia utilized to extract DNA from a mixed pool microti present in human. Recently there are of L3 from each FECRT group. Uniplex PCR many costly procedures are adopted for its reactions were conducted with primer pairs diagnosis. Among all methods Microscopy described in the literature specific for regions detection methods are still the cheapest in the Internal Transcribed Spacer 2 (ITS 2) and fastest method to identify Babesia of Haemonchus sp., Cooperia oncophora, although their sensitivity and specificity Ostertagia ostertagi and Trichostrongylus spp. are limited. Molecular and immunological From 38 L3 cultures where Haemonchus methods have developed and they offer spp was morphologically classified, 20 faster, sensitive and specific methods. These gave an amplicon of 226 base pairs (bp) includes PCR, RT-PCR, DNA Probes, Loop corresponding to the expected size for Mediated Isothermal Amplification (LAMP), this parasite. From 34 samples where Indirect Fluorescent Antibody Test (IFAT), Trichostrongylus spp. was typified, 29 Enzyme-Linked Immunosorbent Assay revelead amplicons of 106 bp reported for (ELISA), Immunochromatography Test (ICT), this genus. From 38 samples where Cooperia Reverse Line Blot Hybridization (RLB). These oncophora was typified, 20 gave amplicons methods are time taking, costly and needs of 192 bp described for the species and out experts. but we need cheap, convenient, of 21 samples where Ostertagia ostertagi portable and sensitive method for detection was typified, four gave amplicons of 124 bp of babesia in blood that should be available expected for this species. at farm which should not require any expert and not time taking .My futuristic approach is The Cohen´s kappa statistics showed a fair that we should go with a kit just cassette of agreement (k value=0.21, p<0.01) between nitrocellulose on which different control lines morphological typification and PCR on of specific dye labelled antibodies against 192 mixed L3 samples. Discriminating by specific antigenic proteins of babesia species nematode, kappa statistics indicated fair .With the buffer, blood drop, lysing agent agreement for Haemonchus contortus (k and simple chromatographic process blood value=0.29; p<0.01), low agreement for along antigenic protein and buffer move on Ostertagia ostertagi (k value=0.12) and the gel and eventually captured by specific Cooperia oncophora (k value=0.14, p=0.10). antibodies and with the help of fluorescence The preliminary results indicate that the of dye we can easily detect which specie is applied PCR protocols successfully identify present in blood of specific animal and human.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 262 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

We can produce its vaccine also if we know report on the students’ ability to correctly the antigenic protein of babesia species so it’s identify microfilaria in the blood samples and another preventive point. It can be done simply if they noticed morphological changes based at farm level and doesn’t need expert person on cryopreservation and fixation used. to check as color lines can be visible easily by anyone and need few drops of blood. PS02.58 Parasitological Procedures, Skills, and Areas of Knowledge Used by Equine PS02.57 Heartworm Microfilaria: Banking the Practitioners in North America Source for Teaching Diagnostic Techniques Dr. Mary Mauldin Pereira1, Dr. Rosina Krecek2, Ms. Sidney Long1, Ms. Jessica Shrake1, Dr. Dr. Anne Conan1, Mr. Christopher Pierson1, Dr. Antoinette Marsh1 Elpida Artemiou1 1The Ohio State University, Columbus, United 1Ross University School of Veterinary States Medicine, Basseterre, St. Kitts, 2Texas A & M College of Veterinary Medicine and Proper diagnosis of Dirofilaria immitis Biomedical Sciences, College Station, United infection is critical to Day 1 veterinarians and States technicians. Antibody-antigen blocking, causing false negatives, confounds some The American Association of Veterinary testing. Therefore, detection of microfilaria is Parasitologists (AAVP) describes the ability an important diagnostic skill. However, finding to proficiently perform and interpret common a supply of blood containing microfilaria diagnostic procedures in parasitology as when scheduled to teach the procedure a core competency. Mastering these skills presents a challenge. To address this issue, requires an understanding of the current we evaluated the use of cryopreserved parasitological procedures, skills, and areas of microfilaria in two standard diagnostic knowledge used in large-animal practice. To tests, Modified Knott’s and carbonate filter investigate this need, a study was designed tests. We also evaluated the impact of by Ross University School of Veterinary environmental friendly 2% acetic acid in place Medicine (RUSVM) where a questionnaire of 2% formalin as a fixation reagent in the involving common procedures was developed assay. The specific aims included determine and completed by 46 equine practitioners. if students could 1) detect the cryopreserved Respondents provided general information on Dirofilaria immitis microfilaria added to practice characteristics identifying the main fresh blood and 2) detect morphological diagnostician, area of their practice, number differences between the microfilaria in of years in practice, number of veterinarians the modified training assays. With this and technicians as well as number of patients information, teaching laboratories could examined per day. Following, participants utilize cryopreserved microfilaria in freshly reported their procedures and frequency on obtained blood in lieu of freshly collected diagnoses of equine helminths, protozoa and microfilaria-containing blood. One aliquot ectoparasites. Descriptive Results indicated of fresh blood was removed, designated as that quick methods requiring minimal a baseline and immediately evaluated using involvement are more likely performed at the two diagnostic techniques. Microfilaria the practice. Specific results regarding the were isolated from whole blood containing diagnosis of equine helminths indicated that JYD or MO microfilaria and cryopreserved practices located in peri-urban areas are using Glycerolyte 57 solution. The microfilaria less inclined to perform fecal flotation using underwent rapid thaw, were spun down centrifugation (p= 0.02). Additionally, the and were added to freshly acquired blood veterinarian is more inclined to perform the from a healthy donor. Students (n = 120) Baermann test (p= 0.01), where the majority were then randomly assigned samples and of diagnosticians perform this test on a yearly fixatives (cryopreserved microfilaria, non- basis (88%). Moreover, the adhesive tape test cryopreserved microfilaria, and either acetic was less likely to be performed in practices acid or formalin fixation) to analyze. We where less than 5 patients were examined

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 263 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

per day (p= 0.02). In practices with 4 or more This surveys reinforce the need to support the veterinarians, skin biopsies to detect mites parasite control guidelines recommended by were more likely to be performed (p= 0.0014). the European Scientific Counsel Companion Though all diagnostic procedures are termed Animal Parasites (ESCCAP) and emphasize common by the AAVP core competencies, the need to examine pets regularly for some results suggest a lack of understanding intestinal parasites. Continued shedding as to what procedures are performed as of parasites in faeces can create heavily well as whether or not they are sent out to contaminated environments, increasing the a diagnostic laboratory. Outcomes of this probability of infection and also of zoonotic study will support parasitology teaching transmission of some parasites. opportunities to further enhance veterinary graduates’ expected core competencies. PS02.60 A One Health Paradigm: Multiple Bovine Infections in the Human-Wildlife- PS02.59 Prevalence of Gastrointestinal Livestock Interface of Lambwe Valley, Parasites of Dogs and Cats in Greater Dublin Western Kenya Area, Ireland Dr. Shewit Kalayou1 Dr. Andres Garcia-Campos1, Mr. Cormac 1International Center of Insect Physiology and Power1, Dr. James O’Shaughnessy2, Mr. Colm Ecology, Nairobi, Kenya Browne3, Ms. Amanda Lawlor1, Dr G McCarthy1, Dr. Emma O’Neill1, Prof. Theo de Waal1 Borne by the tsetse fly, trypanosomosis is a 1University College Dublin, Belfield, Ireland, protozoal diseases complex which has been 2Department of Agriculture, Food and the extensively documented in the Lambwe valley Marine, Backweston Laboratory Campus, since as early as the 20th century. Despite Ireland, 3DSPCA, Rathfarnham, Ireland some level of earlier achievements, the valley lacks long term tsetse and trypanosomosis This study was undertaken to obtain a control plan. At present, there are several snapshot view of the prevalence of canine and complaints by the local community due feline endoparasites in Dublin. Faecal samples to its considerable challenge to livestock were randomly collected from dogs (n=627) productivity and impact the livelihoods. A and cats (n=289) entering a rehoming centre cross-sectional study was undertaken in in Dublin. Samples were analysed by two villages located within 10 km radial distance flotation techniques: one with saturated sugar from Ruma park with the objectives to solution (SG=1.30) and the other zink sulphate estimate the prevalence and to identify risk (SG=1.18) solution, a modified Baermann factors contributing to the occurrence of the technique and faecal smears stained with a disease. A total of 682 blood were sampled Modified Kinyoun’s staining procedure. from local zebu cattle in December 2018 and Surprisingly, 21.7% all dogs and 31.7% of tested using two tests, buffy coat technique all cats sampled harboured at least one BCT and using high-resolution melting (HRM) intestinal parasite. The main parasitic agents analyses. Using the two tests in parallel, the detected were ascarids (15.52 and 30.26%), overall trypanosome infection (Trypanosoma Cystoisospora (3.27 and 3.69%), Giardia congolense, T. brucei, and T. vivax) was 11.6% spp. (6.02 and 1.84%) and lungworms (0.64 and 28.3 %, respectively. Performance of the and 2.08%) in dogs and cats respectively. BCT and HRM was assessed in 682 matching The lungworms identified in dogs were blood samples. The degree of agreement Angiostrongylus vasorum (0.48%) and was found to be fair (75.07%, k= 0.234). T. Crenosoma vulpis (0.16%) while cats were congolense was observed at a rate of 14.8 %, only infected with Aelurostrongylus abstrusus T. brucei at 7.9%, T. vivax at 7.8% and mixed (2.08%). Ascarid eggs and Cystoisospora spp. infection at 2.3%. The putative risk factors oocysts were more often found in young dogs homestead-park distance and village were (p<0.001 and p=0.008, respectively) or cats significantly associated with infection. Our (p<0.001 and p=0.013, respectively) less than analysis showed that 49% of the overall 3 months of age. infection occurred at less than 1.5 km from the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 264 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

park. This is true for T. congolense savannah (2010-19), either as sporadic incidents or as (58%, p ≤ 0.01) and T. vivax (54%, p ≤ 0.01). outbreaks. Occurrence of D. dendriticum was In contrast, more than 67% of the T.brucei documented from 20 counties in NY State sub group infections occurred at ≥ 3 km from (Chenango, Clinton, Columbia, Cortland, the park. Therefore risk-based vector control Delaware, Dutchess, Franklin, Herkimer, interventions should focus on this distance Lewis, Madison, Monroe, Otsego, Queens, range. Further experiments are underway to St. Lawrence, Saratoga, Schoharie, Schuyler, determine what implication of the result exist Tompkins, Ulster, and Wyoming). This includes in the potential spread of infection to human. 17 new counties that were not identified in the 1951 study. Three cases were from counties outside NY State (Grafton-New Hampshire, PS02.61 Retrospective Analysis of Hunterdon-Pennsylvania, and Sussex-New Dicrocoelium Dendriticum (Rudolphi, 1819) Jersey). This study highlights the continued Looss, 1899 Cases Diagnosed During 2010-19 and expanded threat pose by lancet flukes to at the Animal Health Diagnostic Center, New livestock and humans in NE United States. York State, USA

Danielle Maguire1, Holly White1, Manigandan PS02.62 Ehrlichia Canis in Dogs of Mexico: Lejeune1 Prevalence, Incidence, Co–Infection With 1Department of Population Medicine & Rickettsia Parkeri and Factors Associated Diagnostic Sciences, College of Veterinary Medicine, Animal Health Diagnostic Center, Dr. Melina M. Ojeda–Chi1, Prof. Roger Ivan Cornell University, Ithaca, United States Rodriguez-Vivas1, Dr. Maria D. Esteve–Gasent2, Dr. Adalberto A. Pérez de León3, Dr. Joseph J. Dicrocoelium dendriticum (Lancet fluke) is Modarelli2, Sandra L. Villegas–Perez1 an endoparasitic distome with a predilection 1Campus de Ciencias Biológicas y to the bile duct and has a wide host range Agropecuarias. Facultad de Medicina including species of domestic and wild Veterinaria y Zootecnia, Universidad mammals. The lancet fluke was considered Autonoma de Yucatan, Merida, Mexico, a parasite of economic importance in sheep 2Department of Veterinary Pathobiology, and cattle of New York State during the early College of Veterinary Medicine and 1950s. In fact, its occurrence was recorded Biomedical Sciences, Texas A&M University, in six counties (Cayuga, Cortland, Madison, College Station, United States, 3USDA–ARS Oneida, Onondaga, and Tompkins) in central Knipling–Bushland U.S. Livestock Insects NY State, the only region considered enzootic Research Laboratory and Veterinary Pest in the United States in 1951. By late 1980s Genomics Center, Kerrville, United States this was not considered a serious parasite of domestic animals. However, sporadic cases The prevalence, incidence and factors of dicrocoeliosis has been reported, even in associated with Ehrlichia canis infection, as humans, in NY State in the last few decades. well as other Rickettsial infections in dogs of This raised our curiosity to understand the rural communities of Yucatan, Mexico, were current geographical range in the North investigated. A total of 246 dogs were blood Eastern United States. In this study, we sampled and initially screened for pathogens retrospectively analyzed D. dendriticum belonging to the Anaplasmataceae family by cases diagnosed at the AHDC, NY State a quantitative real–time PCR (qPCR) assay, with an aim to understand the geographical to detect Ehrlichia canis, E. chaffeensis, E. spread and the common hosts that it infects. ewingii, Anaplasma phagocytophilum and Diagnosis was based on either detection of Rickettsia rickettsii. Sixty–five dogs were ova by modified Wisconsin fecal floatation monitored and sampled twice 7–8 months test using sugar (1.33 SPG) or through apart. Using the qPCR, 72 positive dogs to E. morphological identification of the flukes. canis were detected (prevalence of 29.26%). A total of 145 cases (92 sheep, 31 cattle, These dogs were also tested by nested PCR 17 goats, 3 bison and 2 llamas) of lancet to detect the same pathogens. None of the flukes were recorded over the last ten years studied dogs were positive to E. chaffeensis,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 265 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

E. ewingii nor A. phagocytophilum by both six months. At the end of this period the PCR assays. The cumulative incidence of animals were euthanized, adult H. contortus E. canis infection was 38.46%. Sequencing were collected, counted and stored for analysis of the nested PCR products revealed RNA extraction. Adult parasite counts were 100% and 98.1% identity of E. canis and R. consistently lower on Moxotó goats with the parkeri, respectively. We found a dog co– exception of 4 animals that had counts similar infected with E. canis and R. parkeri. It is to Saanen goats. Total RNA was extracted concluded that high prevalence and incidence from pools of 20 adult male H. contortus from of E. canis in the dog population of Yucatan the three most resistant (x=243±70 parasites) were detected and age (>3 years old) was the and susceptible Moxotó goats (x=1,981±445 only factor associated with E. canis infection parasites) and from three Saanen̄ goats in dogs. This study presents the first report of (x=2,511±549 parasites). RNASeq̄ was done a R. parkeri active infection in a domestic dog on these samples with 718 million paired in the state of Yucatan, Mexico, presented as a reads̄ generated (160 bases long) by Illumina co–infection with E. canis. sequencing. The paired-end reads where splice-aligned to the H. contortus assembled Research founded by FMVZ–2016–0007, Texas genome (PRJEB506) using HISAT2 and AgriLife and TVMDL’s seed grant, Molecular assembled in 32,460 genes by StringTie. Diagnosis of Zoonotic Tick–Borne Diseases, Using Ballgown, 220 genes were determined and USDA–ARS Project Nos. 3094–32000– to be differentially expressed among the 039–00D and 3094–32000–036–00D. treatments and were blasted in order to determine their identities. These results are currently under analysis and we expect to PS02.63 Transcriptome Analysis of shed some light on the mechanisms related Haemonchus Contortus Infecting Goats from to parasite survival in hosts of different Breeds with Differing Resistance Levels resistance levels.

Dr. Jomar Patricio Monteiro1, Dr. Felipe Rodrigues da Silva2, Edilson Pereira de PS02.64 Cytokine Expression in Calves Freitas3, Janice Araújo Fontenele3, Dr. Jessica Experimentally Infected with Two Theileria Maria Leite dos Santos4, Dr. Marcel Teixeira1, Parva Stocks Dr. Luiz da Silva Vieira1 1Embrapa Caprinos e Ovinos, Sobral, Brazil, Dr. Gloria Patsanza1, Dr. Darshana Morar- 2Embrapa Informática Agropecuária, Leather1, Dr. Nicola Collins1, Dr. Kgomotso Campinas, Brazil, 3Centro Universitário INTA Sibeko-Matjila1 (UNINTA), Sobral, Brazil, 4Universidade 1Vector and Vector-borne Disease Research Estadual do Ceará (UECE), Fortaleza, Brazil Programme, Department of Veterinary Tropical Diseases, University of Pretoria, Haemonchus contortus is the main small Onderstepoort, South Africa ruminant gastrintestinal parasite in tropical areas. Different host breeds present variable Cytokines play a vital role in the immunity and levels of overall resistance against these immunopathogenesis of parasitic diseases. parasites. Moxotó goat breed is locally However, most of the studies on the role of adapted to the Brazilian semi-arid and it cytokines in infection by the tick-transmitted is known for its resistance against these protozoan parasite Theileria parva, were nematodes. Nevertheless, resistant animals performed in in vitro cell lines. Furthermore, still carry gastrointestinal nematodes but there are no comparative studies for different in lower numbers than the susceptible stocks of T. parva even though cattle counterparts. In this study, parasite-free theileriosis caused by T. parva manifests in goats from two breeds (20 Moxotó and different disease syndromes (East Coast 5 Saanen) known to differ in resistance fever and Corridor disease), depending on against nematodes (Moxotó > Saanen) were whether the parasite strain is cattle-derived experimentally infected every two weeks or buffalo-derived. Hence, an in vivo study with the same H. contortus population for was performed to investigate the expression

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 266 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

of cytokines in calves experimentally infected of toll like receptor 2 (TLR2), TLR4, TLR7, with two T. parva vaccine stocks (Katete and interleukin (IL)-10, interferon gamma (IFN- ) Chitongo) used for immunization in Zambia. and (program death ligand) PD-L1 gene mRNA expression of anti-inflammatory expression in LST positive reactions and γ cytokines, IL-10, and IL-4, as well as that paired normal-looking skin of nine infected of pro-inflammatory cytokines, TNF- , IL-2 Ibizan hound dogs. Normal skin from ten and IFN- , were determined using RT-real- seronegative dogs from a non-endemic time qPCR. Notably, both Chitongo- αand area was analysed as a negative control. Katete-infectedɣ calves reacted differently Immune genes expressions were examined to infection, with some showing clinical by quantitative PCR (qPCR) analysis. LST signs (CS) and others not (NCS). The anti- positive reactions presented significant inflammatory cytokine IL-10 expression was upregulation of TLR4, IL-10, IFN- and PD-L1 up-regulated in both CS and NCS animals; and downregulation of TLR7 when compared especially in the Katete-infected CS group. with normal skin of control dogs.γ Moreover, While the expression of IL-4 mRNA was a trend for TLR2 upregulation was observed. generally down-regulated, with significant All transcripts but TLR7 were higher in LST lower expression in the Chitongo-infected CS positive reaction than in paired normal-looking group. The pro-inflammatory cytokines, TNF- skin. The expression profile of immune genes and IFN- , were up-regulated in all groups; in LST positive reactions was similar to that however, IFN- expression in the Chitongo- α previously observed in clinically-lesioned skin infected ɣNCS group was significantly lower of mildly diseased dogs with papular dermatitis compared to otherɣ groups. IL-2 was generally due to Leishmania infantum infection. Our data down-regulated, especially in Chitongo- provide additional support for the important infected NCS group. Overall, these findings role of TLRs in canine leishmaniosis. suggest that T. parva infection influences the expression of investigated cytokines. Similar cytokine expression profiles were generally PS02.66 Human Macrophages Cytokine observed between calves infected with the Response to Dirofilaria Repens Antigens two parasite stocks. However, significant variations were detected in the expression Dr. Magdalena Elżbieta Wysmołek1, levels between the CS and NCS groups; Dr. Ewa Długosz1, Dr. Piotr Bąska1, Dr. suggesting that cytokines such as IL-10, IL-4, Anna Zawistowska-Deniziak2, Dr. Maciej IFN- and IL-2, may have a vital role on the Klockiewicz1, Dr. Marcin Wiśniewski1 disease outcome. 1Warsaw University of Life Sciences, funded ɣ by KNOW (Leading National Research Centre) Scientific Consortium, Healthy Animal - Safe PS02.65 Toll-Like Receptors 2, 4, and 7, Food”, decision of Ministry of Science and Interferon-Gamma, Interleukin 10 and Higher Education No. 05-1/KNOW2/2015, Programmed Death Ligand 1 Transcripts in Warsaw, Poland, 2Witold Stefanski Institute Leishmanin Skin Test Positive Reactions of of Parasitology, funded by KNOW (Leading Ibizan Hound Dogs National Research Centre) Scientific Consortium„ Healthy Animal - Safe Food”, Dr. Laura Ordeix1, Dr. Sara Montserrat-Sangrà1, decision of Ministry of Science and Higher Dr. Pamela Martinez-Orellana1, Dr. Laia Education No. 05-1/KNOW2/2015, Warsaw, Solano-Gallego1 Poland 1Dep. Medicina I Cirurgia Animals, Universitat Autonoma de Barcelona, Bellaterra, Spain Helminths are known for producing immunomodulatory molecules, which allow The leishmanin skin test (LST) is an in vivo them to evade the host immune system. The technique commonly used to evaluate the aim of the study was to investigate the effect Leishmania-specific cellular immune response of somatic antigens from an adult zoonotic in dogs. However, information regarding filarial nematode Dirofilaria repens on the in the local immune response in LST positive vitro production of pro-inflammatory and reactions is scarce. We examined the pattern regulatory cytokines by human macrophages.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 267 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

THP-1 human monocytes were differentiated construction of ever-expanding databases into macrophages and stimulated with of DNA sequences, it has become clear that Dirofilaria repens worm extract (DWE) with parasite morphology is not as accurate a and without LPS. TNF- , IL-6, IL-1 , IL-10 representation of true genetic relationships cytokine concentration was measured in as once believed and has contributed culture medium using ELISAα commercialα kits. to erroneous assignments of generic classifications to many parasites. The genus Macrophages stimulated with DWE released Eimeria is polyphyletic, with members significant amounts of the regulatory IL-10 grouping in several separate clades alongside cytokine comparing to unstimulated cells. The members of Isospora, Cyclospora, Caryospora, treatment had no influence on the secretion and others. Eimeria taggarti, recently of IL-1 , IL-6 or TNF- . The production of described from the prostate of an Antechinus proinflammatory cytokines was upregulated flavipes (yellow-footed antechinus), further only inα THP-1 macrophagesα stimulated with contributes to the polyphyly of the genus LPS. Additional DWE treatment of LPS Eimeria. Based on a partial 18S SSU rDNA stimulated macrophages had no effect on the sequence, E. taggarti groups with reptile- production of pro-inflammatory cytokines. infecting Choleoeimeria and Acroeimeria spp., as well as several Eimeria (s.l.) spp. We Dirofilaria repens extract does not induce completed the sequencing of the nuclear18S the synthesis of pro-inflammatory cytokines SSU rDNA and generated a full mitochondrial and promotes the production of IL-10 genome of E. taggarti; these molecular data immunoregulatory molecule. It might be were used to conduct a phylogenetic analysis suspected that M2 phenotype is induced in to reassess the validity of its taxonomic these cells, promoting the anti-inflammatory assignment. A close relationship amongst E. response. taggarti, Choleoeimeria spp., and Acroeimeria spp. was confirmed. Together, these parasites These findings allow to better understand the form a clade basal to the Eimeria spp., reason for the usual asymptomatic course of sensu stricto. Histological observations of skin dirofilariosis. The next step would be to infected tissues showed that endogenous perform an analysis of the T cell polarization stages are limited to the prostate and that in response to DWE as well as the analysis of sporulation occurs endogenously. Based on immunomodulatory proprieties of identified these phylogenetic data and morphological single D. repens proteins. This may lead to observations, we propose that E. taggarti the establishment of the immunomodulation does not belong in the genus Eimeria, and mechanisms used by the parasite in the merits reclassification. Further, we propose a human host. reconsideration of several of the taxonomic designations within this clade to better reflect the true relationships among its constituents. PS02.67 Phylogeny of a Novel Coccidian Parasite Found in the Prostate of Antechinus Flavipes (Yellow-Footed Antechinus) PS02.68 The First Determination of Eustrongylides Sp. Larvae in the Gulf of Perryn Kruth1, Cassia Michels1, Jemima Amery- Finland and Ladoga Lake Gale2, John R. Barta1 1University Of Guelph, Guelph, Canada, Nadezhda Ryabova1 2Melbourne Veterinary School, Faculty of 1Saint Petersburg State Academy of Agricultural Studies, University of Melbourne, Veterinary Medicine, Saint Petersburg, Russian Melbourne, Australia Federation

Morphological traits, especially those of Abstract: Cases of Eustrongylides excisus sporulated oocysts, have historically been larvae detection in the zander Sander the basis of coccidian phylogeny. With lucioperca caught in the Gulf of Finland and development of molecular tools for assessing Ladoga Lake are described. The dynamics relationships among these parasites and of extensiveness and intensity of invasion

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 268 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

since 2003 are traced, and analysis of the membranous oval structures consistent relationship of these data with fluctuations in with cestode infection. A centrifugal fecal the number of commercial zander in the Gulf flotation test did not detect cestode eggs. of Finland is given. Zander is the paratenic In late November, further needle specimens host for E. excisus and therefore serves as the of cystic lesions were submitted to Antech best indicator of the state of the ecosystem, Diagnostics. Histopathologic findings other food fish (roach, pike, perch etc.) may included extensive necrosis, inflammation also be infected with the fourth-stage larvae and irregular frequently folded PAS-positive of this zoonotic helminth. At the same time hyaline-like membranous material interpreted zander is the most important commercial fish as representing the cyst wall of a larval of both the Gulf of Finland and Ladoga Lake cestode. Additional aspirate material was and a very popular object of game fishing. submitted to the Animal Health Laboratory, Infection of this species with E. excisus has University of Guelph for Echinococcus and a great ecological, economical and medical Taenia PCR testing. A PCR product was significance. Despite the time-distant initial generated using primers specific for E. discovery by the author (as well as numerous multilocularis. Subsequent sequence data consumers of fish products in the markets) of were 100% homologous (344/344 bp) to E. E. excisus larvae in the zander S. lucioperca multilocularis NADH dehydrogenase subunit (2003), this presentation is the first official I gene sequences listed in GenBank. The scientific report confirming the presence parasite masses were considered inoperable of this dangerous nematode in the Gulf of and the dog was placed on albendazole Finland and Ladoga Lake. (10mg/kg) daily. In early February, 2019 the dog’s condition deteriorated and euthanasia was performed. The dog was purchased in PS02.69 Alveolar Echinococcosis in a Dog in Mississippi as a young puppy and brought the Eastern U.S. to Virginia with no history of subsequent travel. This is the first report of alveolar Anne Zajac1, Donald Fairman2, Bridgette echinococcosis in a dog in the United States Wells3, Evan McGee3, Andrew Peregrine4 and the first report of E. multilocularis 1Virginia-Maryland College of Veterinary infection apparently acquired in the Mid- Medicine, Blacksburg, United States, Atlantic region of the U.S. 2Kingdom Animal Hospital, Clear Brook, United States, 3Antech Diagnostics, Fountain Valley, United States, 4Ontario Veterinary PS02.70 Experimental Infection of Domestic College, Guelph, Canada Dogs with Borrelia Turicatae: Clinical Pathology and Cross-Reactivity Echinococcus multilocularis is known to be endemic in Canada and the north central Dr. Jenna Gettings1, Dr. Job Lopez2, Aparna U.S. In early November 2018, an 8-year-old Krishnavajhala2, Brittany Armstrong2, Mr. Alec 36 kg neutered male Labrador Retriever Thompson1, Dr. Michael Yabsley1 was presented to a small animal practice 1University Of Georgia, Athens, United States, in northern Virginia U.S. The owners 2Baylor College of Medicine, Houston, United reported that the dog had recently been States lethargic. Physical exam findings were normal. Radiographs of the abdomen Tick-borne relapsing fever spirochetes are showed loss of detail in the cranial abdomen closely related to Borrelia burgdorferi, the and a possible hepatic mass. An ultrasound agent of Lyme disease and both pathogens performed within a few days of the initial can cause similar clinical signs in dogs. presentation revealed several large hepatic However, the current knowledge of TBRF in masses and multiple smaller masses involving dogs is limited to a handful of case reports the pancreas. Fine needle aspirates of the and so little is known about the clinical hepatic masses were submitted to Antech pathology during the course of acute Diagnostics. The cytologic findings included infection. Despite their similarity, diagnosis inflammation and necrosis with eosinophilic, of TBRF and B. burgdorferi differs in that

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 269 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

TBRF diagnostics are primarily detection of sources of bacteria (manure, waste, wounds) spirochetes using blood smear or molecular and healthy or uninfected animals, including assays, while diagnosis of B. burgdorferi humans, and their habitats. Evaluating and primarily relies on serology. However, reports characterizing the bacterial communities of cross-reactivity in human cases raises the in flies collected from natural habitats will question as to whether dogs seropositive help in understanding their role in harboring to TBRF spirochetes will cross-react on any and disseminating bacteria. In this study, of the several commercially-available assays we used next generation sequencing of designed for B. burgdorferi. bacterial 16S rRNA gene to characterize the gut bacterial communities of adult female TBRF infection dynamics and cross-reactivity house flies collected from three different were evaluated in this study by inoculating environments in Manhattan, KS: agricultural B. burgdorferi-negative dogs with B. (beef cattle feedlot), urban (downtown turicatae, the most commonly identified dumpsters), and mixed environment (business TBRF spirochete in dogs in the United States. located near animal agriculture). Bacterial To describe the dynamic changes in select taxa affiliated to the phylum Firmicutes clinical parameters, temperatures were dominated the bacterial communities in collected daily, platelet counts twice weekly, flies from the agricultural setting while and complete blood counts weekly, during the Proteobacteria was the predominant first two months of infection. Seroconversion phylum in flies from the urban and mixed to B. turicatae was determined by GlpQ environments. In the lower taxonomic Western blot. Samples were tested with level, genera associated with feces or the several commercial and veterinary diagnostic vertebrate gut (rumen-associated microbes laboratory B. burgdorferi-based tests. such as Blautia, Phascolarctobacterium, Anaerovibrio, Clostridiales, Ruminococcaceae, Several samples cross-reacted with Escherichia-Shigella) dominated the commercial and veterinary diagnostic agricultural environment. Importantly, laboratory tests. Practicing veterinarians potential human pathogens including should choose screening and diagnostic Providencia and Aeromonas were tests with care if trying to distinguish dominant in the mixed environment between relapsing fever borreliosis and whereas Enterobacteriaceae, Providencia, Lyme borreliosis. Clinically, the dogs did not Enterococcus and Phascolarctobacterium develop signs of disease, but platelet counts were dominant in urban environment. decreased within the first two weeks post- Principal coordinates analysis showed that inoculation and returned to baseline over the bacterial communities in the house fly the next several weeks. Subclinical infections gut were similar between flies collected from may be present in the population and that the same site, but community composition possibility combined with cross-reactivity of varied significantly across the environments. some tests highlights the need to critically Furthermore, bacterial species richness was evaluate positive tests in the context of an highest in agricultural environment, which animal’s risk of exposure. differed significantly from mixed and urban environments. Similarly, Shannon index was higher in agricultural environment than PS02.71 Bacterial Communities Associated the other two environments. These results with the Gut of Female House Flies Collected demonstrate that the house fly gut harbors From Three Environmental Niches in Kansas complex bacterial communities, including potential human and animal pathogens, and Dr. Dana Nayduch1, Dr. Saraswoti Neupane2 that community composition is strongly 1USDA-ARS, Arthropod-Borne Animal influenced by the environment. Diseases Research Unit, Manhattan, United States, 2Kansas State University, Department of Entomology, Manhattan, United States

Adult house flies serve as a bridge between

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 270 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.72 Use of Barbervax: A Commercial PS02.73 Impact of Shearing on Ectoparasites Haemonchus Contortus Vaccine, in Alpacas in Holstein Cattle over Two Grazing Seasons Cecília José Veríssimo1, Mariana Fogale Michelle Carman1, Grace VanHoy1, Jeffrey Andrade1, Gunta Gutmanis1, Cristiane Lakritz1, Greg Habing1, Kelsey Paras2, Fernandes de Carvalho Fiorin1, Danielle Antoinette Marsh1 Rizatto Balbino1, Mariana Santos Miranda1, 1The Ohio State University College of Bianca Tainá Azevedo1, Jessica Carolina Veterinary Medicine, Columbus, United States, Ribeiro1, Guilherme Favero Luciani1, Luciana 2University of Georgia College of Veterinary Morita Katiki1, Luciandra Macedo de Toledo1, Medicine, Athens, United States Aníbal Vercesi Filho1, Enílson Geraldo Ribeiro1, Rodrigo Giglioti1 Haemonchosis in camelids remains a 1Institute Of Animal Science, Nova Odessa, challenging disease to treat, and prevention Brazil has become increasingly problematic due to widespread anthelmintic resistance. In the tropics, ectoparasites cause extensive Barbervax is an adjuvanted vaccine containing damage to cattle, including the Rhipicephalus natural H-11, H-gal-GP antigens obtained microplus tick, the horn fly Haematobia irritans, from Haemonchus contortus adults via and the larvae of the fly Dermatobia hominis. a proprietary process and solubilized in These parasites are difficult to control because Quil A. This vaccine is approved for use in of its resistance to chemical products. Control Australia after demonstrating its safety and alternatives are required. With the hypothesis efficacy in sheep and goats. The vaccine that the short hair length would reduce the utilizes a mixture of the parasite gut mucosal infestation by the parasites, this work was membrane enzymes including H-gal-GP and carried out. The objective was to know the H11, involved in digesting a blood meal from impact of shearing on natural infestation of the host. This study monitored the efficacy of Holstein cattle, raised on a dairy farm located in the Barbervax vaccine in a group of alpacas. Nova Odessa, State of São Paulo, Brazil. Twenty- In the prior grazing season, three alpacas seven females from four different categories received the vaccine and two remained as were evaluated: lactating cows; lactating heifers; negative controls. These five alpacas were dry heifers and calves, each category raised in then followed during their second grazing different places. These animals were clipped by season after either re-vaccination with a clipper (blade 10) and were evaluated on three Barbervax or received no treatment. To occasions, at 20 days intervals, for infestations further challenge the animals during the by R. microplus, H. irritans and D. hominis larvae. study, each alpaca was dosed with 50,000 Infestation data were transformed into log10 H. contortus larvae. Daily observations (n + 1) to approximate the normal distribution were made. Weekly fecal egg counts (FEC) and were analyzed by means of mixed models were monitored. The re-vaccinated alpacas using time-repeated measures. In this model, developed titers to the H. contortus antigen the category effect, evaluation and their as measured by ELISA. None of the FEC interactions were included, and each variable values went over 500 EPGs. Using terminal was analyzed individually. The mean infestations worm counts, the vaccinated alpacas did not for each category were compared by the demonstrate adult H. contortus worms relative tukey test (p <0.05). The results obtained from to non-vaccinated animals. In conclusion, the the analyzes showed that, in some situations, Barbervax vaccine demonstrated safety when the ectoparasite count was lower in the first used during in the second grazing season in a evaluation, made post shearing, in relation to small group of healthy castrated male alpacas. the other two subsequent evaluations, whereas Despite the large challenge dosing, only one in other situations, the reverse occurred. It was non-vaccinated alpaca demonstrated higher not noticed the impact of the shearing in the EPGs and higher adult H. contortus worm infestation by ectoparasites, since the results burden when compared to the other alpacas were not repeated in the categories (Sponsor: in this trial. Fapesp 2016/19938-7).

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 271 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.74 Rapid Selection of Multiple- 2 ng/ml in all eprinomectin-treated animals Resistance to Eprinomectin and at day 2 post-treatment, indicating that the Benzimidazole in a Dairy Goat Farm lack of effect of this drug was not due to low exposure of the worms to eprinomectin. Lea BordesS1, Nicolas DumontT2, Anne Interestingly, moxidectin treatment was totally Lespine3, Elise Souil4, Jean-Francois Sutra3, effective. This is the first report of multiple- Francoise Prevot4, Christelle Grisez4, Lola resistance to eprinomectin and benzimidazole Romanos4, Aurelie Dailledouze4, Philippe of a Haemonchus contortus population in a Jacquiet4 French dairy goat farm, with moxidectin being 1UMR INRA/ENVT 1225 Ihap, Toulouse, France, a relevant alternative in this case. 2Veterinary practice Gaztainadoia, Saint-Jean- le-Vieux, France, 3INRA-InTheRes UMR 1436, Toulouse, France, 4National Veterinary School PS02.75 Effect of In Vitro Ruminal Digestion of Toulouse, ENVT, Toulouse, France on Solid Formulations of the Nematophagous Fungus Duddingtonia Flagrans Eprinomectin was commercially launched in the late 90’s for lactating cows as a pour- Elizabeth Cespedes-Gutierrez1, Diego Cortes- on formulation with a zero milk withdrawal Rojas1, Jaime Andres Cubides-Cardenas1, Dr. period, and it was recently registered for Martha Gomez-Alvarez1 small ruminants. Given the high prevalence 1Agrosavia, Mosquera, Colombia of resistance to benzimidazoles (BZD) in gastrointestinal nematodes in dairy goats and Pharmacological control of gastrointestinal ewes in France, more and more farmers use nematodes using chemical derived eprinomectin exclusively to treat their animals. compounds represent different problems such We report here the first case of multiple- as anthelmintic resistance, drug residues in resistance to eprinomectin and benzimidazole meat/milk products and alteration in pasture in a French dairy goat farm. A veterinary ecosystem. Biological control of nematodes practitioner noted a poor response to using fungus such as Duddingtonia flagrans two types of eprinomectin treatments (BGMSABV-Df-Col-H-001-2014) is a promising (a topical application and an injectable alternative to overcome these problems in formulation) with high mortality rates. We Colombia. The mode of action of the fungus therefore evaluated the efficacy of several imply the oral ingestion and the transit anthelmintic drugs (eprinomectin, topical through the digestive tract, in the faeces the and subcutaneous routes, moxidectin and fungus generates traps that reduce nematode fenbendazole), using the fecal egg count population. Oral formulations need to be reduction test according to the WAAVP developed to protect fungal spores (conidia guidelines. In parallel, nematode species were and chlamydospores) from the chemical, identified at day 0 and 14 post-treatment physical and microbiological conditions of the after bulk larval cultures, by morphology gastrointestinal cavities. In vitro methods are and real-time PCR. Plasma concentrations of needed as a screening method previous to eprinomectin were analysed by HPLC at day 2 the in vivo assays. The effect of temperature and 5 post-treatment in eprinomectin-treated (39°C), pH and mechanical forces on fungus groups. spores were evaluated in order to correlate the isolated factors with the in vitro ruminal Egg excretions remained high in animals digestion assay. In this work two formulation treated with topical (-16.7% (CI: -237 to 59)) prototypes and a control (dried fungus and subcutaneously (21.5% (CI: -126 to 73)) without any excipient), were submitted to an eprinomectin, and with fenbendazole (-5.8% in vitro assay simulating the ruminal cavity of (CI: -205 to 63)). Haemonchus contortus was ruminants consisting of a mixture of artificial the main species identified by morphology saliva and ruminal fluid under agitation at and real-time PCR before and after treatment 150 rpm and 39 °C. Samples were taken at (97 to 98% of identified larvae), followed 2h, 6h and 12h evaluating the fungal viability, by Trichostrongylus colubriformis. Plasma concentration and in vitro nematophagous concentrations of eprinomectin were above activity. A gradual reduction of the in vitro

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 272 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

nematophagous activity along the time was sequencing. Calves treated with ivermectin observed achieving 69 % for the control, 77 had an 82.5% (95% confidence interval 67.8- % and 79 % for the formulations developed. 90.5) reduction in FEC. The combination These results showed that the aggressive treatment of injectable ivermectin and oral conditions in the ruminal cavity could affect fenbendazole had a 100% efficacy in reducing the fungal ability to capture the nematodes FEC. In the ivermectin group, and the in vitro method could be employed as was the predominant parasite species a selection tool for formulations. before treatment; however, 혖혴혵혦혳혵혢혨혪� was혰혴혵혦혳혵혢혨혪 predominant post-ivermectin treatment. There was an increase in the혊혰혰혱혦혳혪 relative� 혰혯혤혰혱혩혰혳 � PS02.76 Ivermectin-Resistant proportion of in calves in Feedlot Cattle from Western following ivermectin treatment compared to Canada 혊혰혰혱혦혳혪� that of pre-treatment. 혏혢혦혮혰혯혤혩혶 This� 혱혭혢혤혦혪 study confirmed 혖혯혤혰혱혩혰혳혢 resistance to ivermectin Mr. Eranga De Seram1, Dr. ElisabethRedman2, while was ivermectin Ms. Camila de Queiroz2, Dr. John Gilleard2, sensitive.혊혰혰혱혦혳혪� 혰혯혤혰혱혩혰혳혢 The increase of Dr. John Campbell1, Dr. Colleen Pollock3, Mr. following혖혴혵혦혳혵혢혨혪 ivermectin� 혰혴혵혦혳혵혢혨혪 treatment is concerning Samantha Ekanayake1, Dr. Fabienne Uehlinger1 because it can cause devastating혏혢혦혮혰혯혤혩혶 losses� 혱혭혢혤혦혪 in 1University of Saskatchewan, Saskatoon, beef cattle operations. In addition, this study Canada, 2University of Calgary, Calgary, identifies regional, accurately phenotyped, Canada, 3Merck Animal Health, Kirkland, ivermectin-resistant parasite populations for Canada future molecular studies.

Gastrointestinal nematode resistance against commonly used anthelmintics in cattle is PS02.77 Resistance of Sheep increasingly reported; however, investigations Trichostrongylids Against Moxidectin in in Canadian beef operations are limited. Austria Therefore, we determined the efficacy of currently used anthelmintic products in Dr. Barbara Hinney1, Untersweg Florian1, Prof. feedlot cattle from western Canada. One Dr. Anja Joachim1 hundred and fifty-six auction market-derived, 1Institute for Parasitology, Vetmeduni Vienna, weaned, fall-placed steer calves were Vienna, Austria randomly allocated to two treatment groups: injectable ivermectin; injectable ivermectin Anthelmintic resistance (AR) in sheep combined with oral fenbendazole. Each group nematodes is increasingly reported contained replicates of 6 pens and 13 animals worldwide. Few studies have been per pen. Anthelmintics were administrated conducted in Austria and while AR against according to individual body weights and benzimidazoles (BZ) seem to be frequent, manufacturers’ recommendations. Fecal egg AR against macrocyclic lactones (ML) has counts (FEC) were obtained from each calf not been clearly identified yet. As severe before and 14 days post-treatment using a treatment failures were increasingly reported modified Wisconsin floatation technique. from sheep flocks we aimed to monitor the To determine the anthelmintic efficacy, efficacy of BZ and ML by fecal egg count we calculated the percentage reduction in reduction tests (FECRT) in selected farms. In post-treatment FEC from pre-treatment total 11 farms were examined, 3 from Styria, FEC using the package “eggCounts” in R 8 form Tyrol. Overall, 500 animals were statistical software. Anthelmintic resistance included. At the first farm visit feces was was determined when the reduction in mean taken and examined by Mini-Flotac (detection FEC was <95% and the upper and lower limit EpG= 5). Animals with an EpG>100 were 95% confidence intervals were <95 and included. A larval culture was set up from <90%, respectively. Relative proportions of pooled fecal samples before treatment and gastrointestinal nematode species in calves in case of a positive FEC also after treatment. pre- and post-treatment was determined Animals were treated with Moxidectin by deep amplicon ITS-2 rDNA nemabiome (Cydectin®) (MOX, group 1) Fenbendazol

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 273 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

(Panacur® ) (BZ, group 2) or left untreated study was to assess the resistance profile as controls (group 3). All three groups could of SCFT infesting livestock at a farm in the be tested on one farm. Moxidectin was Department of Arauca, Colombia, through applied on ten farms, Fenbendazol on three. bioassays and molecular techniques. Results FECRT indicated a reduced sensitivity of from multiple Adult Immersion Tests (AIT) trichostrongylids against Mox on two farms showed complete lack of deltamethrin (78%, 79%) and against BZ on two farms (48%, efficacy at concentrations of 2 and 4x above 89%). The predominant nematode genera therapeutic levels of 50 ppm. The Larval before treatment were Haemonchus spp., Immersion Test (LIT) with deltamethrin followed by Trichostrongylus spp. and the confirmed the high resistance status (RR Chabertiinae. After treatment predominantly = 241.6). At label concentrations, the Haemonchus spp. and to a lesser extent organophosphorus chlorpyrifos (312 ppm) Trichostrongylus spp. were found. The had an intermediate value efficacy of 65-75%, apparent lack of efficacy of the ML in sheep whereas ethion (622 ppm) maintained 100% flocks with visible clinical consequences efficacy. With the (LIT), the LC50 (6.34 ppm) indicates the urgency of implementing and LC99 (22.79 ppm) for ivermectin were sustainable control practices in Austria. not different to a susceptible SCFT strain, and 5-6 fold lower than values published before showing lack of field efficacy. Due PS02.78 Acaricide Resistance Profile of to complete lack of efficacy against SCFT, Rhipicephalus Microplus From Arauca, pyrethroids were no longer used by the Colombia producer in this farm. Ethion was the main acaricide employed with alleged good SCFT Dr David Villar1, Mr Arlex Rodríguez-Durán2, control. A PCR-RFLP technique to investigate Mr Felipe Bossio1, Dr. Robert Miller3, Dr. the presence of a carboxylesterase mutation Guilherme Klafke4, Dr. Adalberto Pérez de associated with pyrethroid resistance revealed León5, Dr. Jesús A. Cortés-Vecino2, Dr. Jenny J. a mixture of homozygous wild-type (n=4), Chaparro-Gutiérrez1 heterozygous (n=13), and homozygous (n=10) 1Grupo de Investigación CIBAV, Escuela de mutant genotypes, although SCFT showed Medicina Veterinaria, Facultad de Ciencias a resistant phenotype against deltamethrin. Agrarias, Universidad de Antioquia, Thus, different resistant mechanisms to Medellín, Colombia, 2Grupo de Investigación pyrethroids may be involved including Parasitología Veterinaria, Departamento de mutations in the para-sodium channel. These Salud Animal, Facultad de Medicina Veterinaria results complement reports from other y de Zootecnia, Universidad Nacional de regions of Colombia where a complete lack Colombia , Bogotá D.C., Colombia, 3USDA-ARS of efficacy occurs towards pyrethroids. Best Cattle Fever Tick Research Laboratory and practices need to be developed regarding the Veterinary Pest Genomics Center, Edinburg, use of ethion as the only acaricide affording United States, 4Instituto de Pesquisas complete SCFT efficacy across different parts Veterinarias Desiderio Finamor, Eldorado of Colombia. do Sul, Rio Grande do Sul, Brasil, 5USDA- ARS Knipling-Bushland US Livestock Insects Research Laboratory and Veterinary Pest PS02.79 The Prevalence of Anthelmintic Genomics Center, Kerrville, United States Resistance in Gastrointestinal Nematodes of Beef Cattle in Uruguay Rhipicephalus microplus, commonly known as the southern cattle fever tick (SCFT), Dr. America Mederos1, Dr. Celeste Serrano1, prevents raising pure Bos taurus cattle breeds Dr. Laura Rinaldi2, Dr. Georg von Samson- in Colombia because it feeds on blood and Himmelstjerna3 transmits pathogens. Intense chemical 1Instituto Nacional De Investigacion treatments with synthetic acaricides as Agropecuaria, Tacuarembo, Uruguay, the only control method selected for SCFT 2University of Naples, Department of populations that are resistant to multiple Veterinary Medicine and Animal Production, classes of acaricides. The objective of this Naples, Italy, 3Free University of Berlin,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 274 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Parasitology and Tropical Veterinary Medicine PS02.80 Comparative Efficacy of Doramectin Institute, Berlin, Germany 1% and Closantel 10% + Albendazole 5% in Goats from the City of General Artigas, The aim of this study was to determine the Canindeyú Department, Paraguay prevalence of anthelmintic resistance (AR) in gastrointestinal nematodes (GIN) in beef Perla Gómez1, Dr. Antonio Rodríguez2, Dr. cattle farms under extensive production Laura Portillo1, Dr. Jorge Miret1 systems in Uruguay. A cross-sectional study 1Facultad de Ciencias Agropecuarias y was designed and the sample size (n=35-40 Ambientales (FCAA). Universidad Nacional farms) estimated under the assumption of de Canindeyú (UNICAN), Curuguaty, 20% prevalence of AR; 5% allowable error Paraguay, 2Centro de Diagnóstico Veterinario and 95% confidence interval. The farms were (CEDIVEP), San Lorenzo, Paraguay conveniently selected mainly in the North part of the country, based on willingness to Small ruminant like goats are susceptible to participate. Drug efficacy was evaluated using helminth infection so generally the control the in vivo Fecal Egg Reduction Test (FECRT) of gastrointestinal nematode relies primarily for the following drugs: Ivermectin (IVM, on antihelminthics administration by oral 200 g/kg bodyweight), Levamisole (LEV, o subcutaneous route. The objective of 7.5mg/kg bodyweight), Ricobendazole (RBZ, this study was to determine the efficacy of 4mg/kgµ bodyweight) and Fenbendazole doramectin and closantel in 38 Boer and (BZ, 5mg/kg bodyweight). Additionally, BZ mixed goats from Yvy Kui farm, in General efficacy was evaluated by the in vitro Egg Artigas City located at the Canindeyú Hatch Test (EHT). At each farm on Day 0, 15 Department in Paraguay. Two group animals were assigned randomly to each of were used. Group 1: Received doramectin the treatment groups (T) and one group was (Dectomax®, Zoetis, Argentina), at the left untreated (C). For each group, individual recommended dosage of 200 g/Kg by fecal samples were collected at baseline and subcutaneous injection) and Group 2: received day 14 and analyzed by the Mini-FLOTAC closantel 10%, 10 mg/kg plus albendazoleμ 5% technique. The efficacy was obtained using 5 mg/kg (Microtel®, Laboratorios Microsules the formula described by Dash [1-(( C0/C14)/ Uruguay, delivery orally). Fecal samples (T14/T0))]*100 and a threshold of <95% was were collected at the time of treatment and the criteria to declare AR positive A pool 7, 14 and 21 days later. The fecal egg count of samples for each farm was kept under was performed by the McMaster technique anaerobic conditions for EHT. Coprocultures (Coles et al., 1992) and the fecal culture and were done for each treatment group. three stage larvae genus identification (L3) From 27 farms sampled until present, results according to Fiel et al., 2011. The fecal culture from the FECRT indicated a proportion of before the treatment showed the infection by 100%, 29.6%, 22.2% and 4.7% presenting Cooperia sp. (45%), Haemonchus sp. (32%), lack of efficacy for IVM, LEV, RBZ and BZ, Ostertagia sp. (18%) and Nematodirus sp. respectively. The EHT showed a range (5%). In both groups 5/19 (26,3%) goats EC50 values of 0.01 g/ml-0.088 g/ml, showed a reduction of 90% in the fecal egg indicating no evidence of GIN resistance count at 14 days post treatment and the to Thiabendazole (Thresholdµ >0.10µ g/ml). same percentage was registered at 21 days Cooperia spp. was the main GIN genera post treatment in 7/19 (36,8%) goats of both identified as resistant in all drug groups.µ groups. An average of 13/19 (68.4%) goats The preliminary results of the present study of the tested animals in both groups did not demonstrated the widespread prevalence of showed any reduction in their egg counts. AR in GIN of beef cattle in Uruguay as already This study showed the high resistance to reported for sheep. these antihelminths molecules in goats and major studies should be carried out in order to evaluate the real impact of the antihelminth resistance in these animals so appropriate strategies can be developed to address this problem.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 275 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.81 Pomegranate (Punica Granatum) (protein 5.5%, casein 4.1%, lactose 4.3%, fat Aqueous Extract for Control of 8.3%). Although results showed a discrete Gastrointestinal Nematodes in Sheep anthelmintic efficacy, the benefits derived from not using synthetic drugs and the quali- Fabio Castagna1, Antonio Bosco2, Rosalia quantitative increase in milk production Crupi3, Valeria Maria Morittu1, Prof. Domenico indicate the use of the mixture as useful and Britti1, Vincenzo Musella1 promising in control strategies against GIN. 1Department of Health Sciences, University of Catanzaro Magna Graecia, Catanzaro, Italy, 2Department of Veterinary Medicine PS02.82 Efficacy Evaluation of a Formulation and Animal Production, University of Naples with Ivermectin 1% w/V Plus Clorsulon 10% Federico II, Eboli, Italy, 3Department of w/V (Ivomec® F) in Cattle Naturally Infected Chemical, Biological, Pharmaceutical and with Fasciola Hepatica in Brazil Environmental Science, University of Messina, Messina, Italy Luis Pelissoni1, Isabela Martins2, Fabio Scott2, Gabriela Oliveira2, B Avelar2, Cristiano Helminth infections, and in particular Nascimento3, Debora Sartor1, Andre Cutolo1 Gastrointestinal nematode (GIN) are amongst 1Boehringer Ingelheim Animal Health Do Brasil the most important production-limiting Ltda., Paulínia, Brazil, 2Universidade Federal pathogens of grazing ruminants globally. The Rural do Rio de Janeiro, Seropédica, Brazil, negative impact of GIN on livestock farms 3Convolution, Ribeirão Preto, Brazil is further exacerbated by the escalating spread of anthelmintic resistance (AR). Fasciola hepatica is a trematode that Phytomedicine has been used by farmers to parasitizes the liver of several species treat parasitism and improve performance including ruminants and is responsible for of livestock, however, scientific evidence a decrease in animal welfare and significant on the anti-parasitic efficacy of most plant economic losses in cattle and sheep. In Brazil, products is limited. Scientific validation of the area with higher prevalence of bovine the anti-parasitic effects is necessary prior to fasciolosis is the Southern region, however their adoption as a novel method for parasite the disease has also been noted in the states control. The aim of this study was to evaluate of Rio de Janeiro, São Paulo, Espírito Santo, the anthelmintic efficacy and benefits on the Minas Gerais and Goias. The treatment of milk production of the natural extract rich Fasciola hepatica infection is mainly done in tannin present in the peels and seeds of through the use of chemical substances. This the pomegranate (Punica granatum). The study aimed to evaluate the efficacy of a research was conducted in sheep naturally single subcutaneous injection of a commercial infected by GIN. 30 sheep were selected and formulation with ivermectin 1% plus clorsulon divided into 2 groups of 15 sheep each: TG, 10% w/w (Ivomec® F) in bovines naturally treated orally at single-dose with 50 ml of infected with Fasciola hepatica. The study aqueous extract 21 days before calving; CG, was conducted on the facilities of the Federal untreated. On Days 7, 14 and 21, individual Rural University of Rio de Janeiro State faecal samples were collected to evaluate and the animals were obtained from Alegre the Faecal egg count (FEC) using FLOTAC city, state of Espírito Santo. Twenty bovines technique and FEC reduction (FECR) on the (age = 9.5 ± 4.1; weight = 380.3 ± 59.8) were different days. The formula used to evaluate allocated pre-treatment to blocks based on was 100x(1-[epgTG/epgCG]). The milk decreasing arithmetic means EPG counts production was measured 8 times every 7 for Fasciola hepatica eggs, performed on days starting from beginning of lactation. The Days -7, -5 and -3 and divided in two groups results of epg mean and FECR (%) were: TG- (Control and Treated) of 10 animals each. D0: 460; D7: 264 (55.7%); D14: 298 (53.8%); EPG means pretreatment for control and D21: 392 (46.1%); CG-D0: 450; D7: 596; D14: treated group was 8.7 ± 5.2 years and 8.8 ± 646; D21: 728. Regards the milk was observed 5.5 kg, respectively. Animals from Treated an increase average over the entire lactation Group received (Day 0) a single subcutaneous both quantitatively (15.5%) and qualitatively injection of Ivomec® F (1mL/50kg bw). On

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 276 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Days 19 and 20 the animals were euthanized parameters reflecting the rate and extent and the livers were removed and sliced into of IVM absorption, indicate the existence 2cm pieces to recover adult specimens of of some differences among preparations Fasciola hepatica on bile ducts. All flukes in terms of pharmaceutical behaviour. The recovered were counted. No specimens relationship between the plasma kinetic of Fasciola hepatica were recovered from profiles of the different formulations and the Treated Group animals, while in Control Group “theoretical threshold” to obtain an optimal mean fluke number observed per animal was efficacy against ticks was calculated using 12.60 ± 8.64. In conclusion, Ivomec® F efficacy the period of time during which ivermectin was 100% on the treatment of Fasciola concentrations were above 10 ng/mL. hepatica in bovine naturally infected. Formulations A, B and D remained above 10 ng/mL for a longer period of time compared to Formulations C, E and F (a highest dose PS02.83 Comparative Ivermectin Plasma rate of 800 g/kg was used for formulation Concentration Profiles After Subcutaneous D). The tested Long-Acting ivermectin Administration of Different Long-Acting formulationsµ showed slight differences in Formulations to Cattle their absorption patterns, which was reflected in the observed plasma pharmacokinetic Paula Leite1, Dr. Evelise Fonseca1, Dr. Sandra behavior. Formulations A and B showed the Fernandez1, Reinaldo Imato1, Bergson Oliveira1, best performance from the pharmacokinetic Prof. Luis Alvarez2, Dr. Mariana Ballent2, Prof. point of view, showing initial higher ivermectin Guillermo Virkel2, Ms. Paula Domínguez2, Dr. concentrations followed by sustained plasma Candela Canton2, Prof. Carlos Lanusse2, Prof. concentrations above 10 ng/mL for more than Adrian Lifschitz2 40 days that may be relevant to obtain an 1Ceva Saúde Animal Ltda., Paulínia, Brazil, optimal efficacy against ticks. 2Laboratorio de Farmacología, Centro de Investigación Veterinaria de Tandil (CIVETAN), CONICET-CICPBA-UNCPBA, Facultad de PS02.84 Effect of Monepantel on Ciencias Veterinarias, UNCPBA, Tandil, Gastrintestinal Nematodes Inefection and the Argentina Infulence on Gestant Sheep

The pharmacokinetic behaviour and systemic Luciana Katiki1, Karen Tiemi Akashi3, Eveline availability of ivermectin were compared Henrique dos Santos3, Jessica Priscila da following subcutaneous application of Paz3, Luciano Antunes Barros3, Poliana Araujo six different Long-Acting formulations Pacheco1, Cecilia Jose Verissimo1, Claudio A to cattle. Forty-two healthy calves were Massamitsu Sakamoto2 randomly allocated into six experimental 1Institute of Animal Science (IZ/APTA), Nova groups (n=7). Animals in each group were Odessa, Brazil, 2Federal University Fluminense, subcutaneously treated with one ivermectin Niteroi, Brazil, 3State University of Maringa, formulation at label recommended dose rate Umuarama, Brazil (Formulation A= 700 g/kg; B= 700 g/ kg; C= 630 g/kg, D= 800 g/kg; E= 700 One of the main impasses in sheep breeding g/kg; F= 630 g/kg).µ Blood samplesµ were is the gastrointestinal nematodes (GN) caused collected overµ 120 days post-treatmentµ (14 by the high morbidity and mortality of the samplingµ times).µ Ivermectin concentrations animals. Several anti-helminthic molecules in plasma were measured by HPLC. Complete have been used to control GN, especially pharmacokinetic analysis were performed during the critical period of sheep peripartum. for all ivermectin preparations. The mean However, the indiscriminate use of drugs peak plasma concentration (Cmax) and favors the development of resistance by the area under the concentration vs time helminths. Monepantel belongs to the new curves (AUC, drug exposure) obtained for class of aminoacetonitrile derivatives and is each formulation were compared following indicated for the treatment and control of dose rate normalization. The statistically multiresistant GN in sheep. The objective significant differences observed in the kinetic of this study was to evaluate the influence

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 277 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

of the peripartum period on the helminth This work evaluated the in-vivo and in- parasite load, the anthelmintic efficacy of vitro pharmaco-chemical interaction and Monepantel during the gestational period of the in-vivo efficacy of the combination the sheep and whether its use generates any of albendazole (ABZ) with a phenolic influence on the development of gestation natural monoterpene, thymol (TML), in and the health of newborn lambs. Thirty- lambs naturally infected with resistant two pregnant sheep were divided into two gastrointestinal nematodes. Thirty (30) lambs groups (G) of 16 animals each, according were allocated into three (3) experimental to egg count per gram of feces (EPG) at groups. Each group was treated orally with the 4th week of gestation (WG). G1 was either ABZ (5 mg/kg), Thymol (150 mg/kg, treated with Monepantel at the 8th and 16th twice every 24 h) or the co-administration WG; G2 was untreated (control). Stool were of both compounds. Blood samples were collected for EPG at 4th, 8th, 12th, 16th and collected between 0 and 51 h post-treatment 20th WG and two weeks after lambing. At and TML, ABZ and its metabolites were birth, all newborns were clinically examined. determined by HPLC. Individual faecal Monepantel did not present side effects samples were collected at days -1 and 14 in the gestation of the sheep, and all the post-treatment to perform the faecal eggs offspring were born healthy. It was also count reduction test. Additionally, the effect verified that the first 10 days after delivery of TML on the metabolic sulphoreduction and correspond to the period with the highest the sulphonation of ABZ sulphoxide was in- parasitic infection. In the present study, the vitro assessed using ruminal content and liver treatments with interval of 63 days showed microsomes, respectively. No changes on the efficient reduction of EPG during peripartum. pharmacokinetic behavior of ABZ sulphoxide In addition, Monepantel did not cause clinical were observed in the presence of the natural and gestational changes in the offspring, the product (TML). In contrast, the ABZ sulphone two treatments performed at the 8th and 16th Cmax and AUC were lower (p<0.05) in the week of gestation had satisfactory efficacy in co-administered animals (0.16±0.07 g/mL the control of GN in pregnant sheep. y 3.63±1.21 g.h/mL) compared with those that received ABZ alone (0.45±0.15 µg/mL and 9.50±2.84µ g.h/mL). TML was detected PS02.85 Combination of Bioactive in the bloodstream between 1 and 51µ h Phytochemicals and Synthetic Anthelmintics: post-treatment,µ which indicates the time of In Vivo and in Vitro Assessment of the target nematodes exposure to the bioactive Albendazole-Thymol Association monoterpene. However, the in-vivo efficacy of TML was 0% and the presence of TML did not Victoria Miro1, Sonia Luque2, Mercedes increase the efficacy of ABZ. The presence Lloberas2, Livio Costa-Junior3, Carlos Lanusse1, of TML inhibited significantly (P< 0.05) the Guillermo Virkel1, Adrian Lifschitz1 ruminal sulphoreduction and the hepatic 1Centro de Investigación Veterinaria de Tandil sulphonation of ABZ sulphoxide. In-vivo (CIVETAN)(CONICET-CICPBA-UNCPBA), pharmaco-parasitological studies are relevant Facultad de Ciencias Veterinarias, Universidad to corroborate the adverse kinetic/metabolic Nacional del Centro, Tandil, Argentina, interactions and the efficacy of bioactive 2Laboratorio de Parasitología, Instituto natural products combined with synthetic Nacional de Tecnología Agropecuaria (INTA), anthelmintics. Balcarce, Argentina, 3Centro de Ciências Biológicas e da Saúde, Universidade Federal do Maranhão, Maranhão, Brazil

The search of novel strategies is an urgent need considering the widespread increasing of anthelmintic resistance in livestock. Bioactive phytochemicals may contribute to improve parasite control by enhancing the effect of existing anthelmintic drugs.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 278 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.86 Evaluation of the Ethnoveterinary PS02.87 Emerging and Reemerging Zoonotic Potential of Picralima Nitida (Apocynaceae) Parasitic Diseases of Companion Animals in as an Anthelmintic India

Dr. Terry Nzeakor1, Dr. Francis Obi1, Dr. Sandra Dr. Arumugam Samgaran1, Dr. Deka Neelakshi1 Nzekwe1, Dr. Martin Omeje1, Dr. Kalu Idika1, 1Madras Veterinary College, Tamilnadu Prof. Chukwunyere Nwosu1, Prof. Romanus Veterinary and Animal Sciences University, Ezeokonkwo1 Chennai, India 1University Of Nigeria, Nsukka, Enugu, Nigeria There are well recognised zoonotic infectious Aim of the study: The study evaluated the diseases transmitted from companion animals potentials of P. nitida as an anthelmintic. to human beings across the globe and Materials and methods: Up and down method among the various pathogens responsible for of acute toxicity was used to determine the this zoonoses, parasites play an important acute toxicity of the ethanol seed extract of role. Pet animals play a significant role to Picralima nitida (ESEPN) up to 2,000 mg/kg the social, physical, mental and emotional body weight given orally. The anthelmintic development of kids, and are a companion efficacy of ESEPN was investigated in to the elderly people and these pets are vitro using the egg hatch assay and in almost a family member in most of the vivo using thirty adult male albino mice houses. Most of the cases or instances, the randomly divided into 6 groups (A–F) of 5 disease due to such zoonotic parasites goes mice each and experimentally infected with undiagnosed or unnoticed due to lack of Heligmosomoides bakeri (with the exclusion awareness or knowledge. The presence of of mice in Group A which was the negative the gastro intestinal parasites in pet animals control group). Graded doses of ESEPN leaves a major problem in transmitting the (250mg/kg, 500mg/kg and 1,000mg/kg) and disease to human beings and cause severe Albendazole (25mg/kg) were administered to morbidity. The transmission is possible to mice in groups D – F and C respectively. The human beings by ingestion of infective stages body weight, packed cell volume, erythrocyte, such as oocysts, cysts, spores, ova, larva or total leucocyte and faecal egg counts (FEC) larval stages or through consumption of raw were assessed. or undercooked meat with infective larval stages in tissues. Companion animals like Results: No death or sign of toxicity was dogs and cats act as reservoir host for a observed in the mice following acute toxicity number of pathogens of parasitic zoonoses, assay. LC50 of 0.120 and -1.557 for ESEPN viz., toxoplasmosis, giardiosis, toxocarosis and and Albendazole respectively was obtained ancylostomosis. Their roles in transmitting following a Probit log regression analysis of human infections have been recognized the percentage egg hatch inhibition. A pre- worldwide. An integrated multidisciplinary patent period of 7±3 days was observed. ‘One Health’ approach is the need of the Anaemia, weight loss and leucocytosis hour to prevent transmission of zoonotic were observed following infection. A dose parasites, necessitating collaboration dependent increase to near pre-infection between veterinary and medical scientists, values of the body weight and haematological policy makers and public health officials. The parameters was observed following treatment interaction amongst veterinary and human with either albendazole or ESEPN. A drop medicine with clinicians, researchers, agencies in FEC was observed till days 9 and 10 post and governments together will benefit treatment for ESEPN and Albendazole domestic as well as wild animals, human respectively however; none was able to clear health and the global environment in general. the helminth infection.

Conclusion: Despite promising in vitro anthelmintic activity of ESEPN, its in vivo was poor. Thus ESEPN should be exploited for other uses.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 279 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS02.88 Plasmodium Knowlesi and Other Poster Session 03 Malaria Parasites in Long-Tailed Macaques from the Philippines July 10, 2019, 10:00 – 16:30 Exhibit Hall A, Level 1 Mr. Lief Erikson Gamalo1, Dr. Judeline Dimalibot1, Ms. Khamisah Abdul Kadir2, Dr. Vachel Gay Paller1, Prof. Balbir Singh2 PS03.01 Treatment of Tritrichomonas 1University of the Philippines Los Baños, Foetus in a Cat Colony with Pre-existing Laguna, Philippines, 2Universiti Malaysia Neurological Disorders Sarawak, Kota Samarahan, Malaysia Dr. Barbara Hinney1, Isabelle Christen1, Macaca fascicularis (long-tailed macaque) Stéphanie Jähne1, Andrea Markl3, Stefanie is the most common species of macaque Gasibauer2, Prof. Anja Joachim1, Dr. Priv.-Doz. in Southeast Asia and the only species of Frank Künzel2 monkey found naturally in the Philippines. The 1Institute for Parasitology, Vetmeduni Vienna, species is the natural host for the zoonotic Vienna, Austria, 2Clinical Unit of Internal malaria parasites, Plasmodium knowlesi and Medicine Small Animals, Vetmeduni Vienna, P. cynomolgi and for the potentially zoonotic Vienna, Austria, 3Clinical Unit of Obstertric, parasite, P. inui. Other Plasmodium species Gynaecology and Andrology, Vetmeduni such as P. coatneyi, P. simiovale and P. fieldi Vienna, Vienna, Austria are also natural parasites of M. fascicularis. The aims of this study were to identify and Aim of the study was to eliminate the determine the prevalence of Plasmodium diarrheal agent Tritrichomonas foetus from species infecting wild and captive long-tailed a cat colony with co-existing morbidity. macaques from the Philippines. A total of 95 Seventeen Maine-Coon cats were confiscated blood samples from long-tailed macaques for animal welfare reasons and kept in an were collected from three locations; 30 at the animal shelter. Six cats showed neurological National Wildlife Rescue and Rehabilitation disorders before treatment. Liquid diarrhea Center (NWRRC) in Luzon, 25 at the Palawan was observed in three cats, soft faeces in Wildlife Rescue and Conservation Center four. Fourteen cats were tested positive for (PWRCC) in Palawan and 40 from Puerto T. foetus by PCR. All cats were treated with Princesa Subterranean River National Park ronidazol (Ridzol®, 30 mg/kg PO q24h) for (PPSRNP) in Palawan. The Plasmodium spp. 14 days. Two weeks after treatment T. foetus infecting the macaques were identified using could not be detected by PCR anymore; species-specific nested PCR assays on DNA however, three to five weeks later three cats extracted from these blood samples. All 40 were again positive for T. foetus. The success of the wild macaques from PPSRNP and 5 rate for elimination of the pathogen within the of 25 captive macaques from PWRCC were observation period of 8 weeks post treatment Plasmodium-positive, while none of the 30 was 79%. Liquid diarrhea could be controlled captive macaques from the NWRRC had any in all three cats. During treatment nine cats malaria parasites. Overall, P. inui was the most showed neurological disorders at least once. prevalent malaria parasite (44.2%), followed In five of cats that showed neurological by P. fieldi (41.1%), P. cynomolgi (23.2%), P. disorders before treatment a minor coatneyi (21.1%), and P. knowlesi (19%). Mixed impairment of clinical signs (coordination species infections were also observed in 39 problems, head tilt) was observed. As of the 45 Plasmodium-positive macaques. neurological signs occurring during treatment Wild long-tailed macaques from the island were similar to the clinical signs observed of Palawan, the Philippines are infected with before treatment it could not be determined P. knowlesi, P. inui, P. coatneyi, P. fieldi and if these were side effects of the ronidazol P. cynomolgi. The presence of these simian therapy. Three cats showed neurological Plasmodium parasites, especially P. knowlesi disorders for the first time during treatment and P. cynomolgi in the long-tailed macaques (slight head tremor and slight ataxia). In in Palawan presents risks for zoonotic conclusion, due to possible side effects transmission in the area. of ronidazol in cats a treatment of feline

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 280 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

tritrichomonosis should always be decided Bw of this new formulation containing 0.5% on a risk-benefit assessment. Nonetheless, Eprinomectin, 5% Diflubenzuron (Virbac) systemically ill animals should not generally were poured on the back of group T animals, be excluded from treatment, especially when from the base of their horns to the base of they are living in animal shelters where a good their tails. EPG counts and coprocultures were health status is a prerequisite for adoption. performed on days D-7, D-2, D+7, D+14, D+21, Because the affected cats were all closely D+28 and D+ 35. related, we hypothesize that a high coefficient of genetic relation might be a significant risk On D-7, D-2 the average EPG counts in factor for the development of tritrichomonosis arithmetic mean were 695 and 790 in groups in cats. C and T, respectively. Coprocultures at D-7 included Cooperia spp (38%), Haemonchus spp (32%), Oesophagostomum spp (27%), PS03.02 Efficacy of a New 0.5% Trichostrongylus spp (3%), respectively. In Eprinomectin, 5% Diflubenzuron-based Pour- group T, EPG counts decreased significantly On Formulation against Gastro-Intestinal (p <0.05) in comparison with group C for Nematodes in Cattle Population under Field all time points from D+7 until D+35, and, in Conditions in Brazil arithmetic means by 91.2%, 95.60%, 89.5%, 89.5 and 74.5%, respectively. Dr. Fabio Barbour Scott2, Dr. Gabriela Ferreira de Oliveira2, Dr. Barbara Rauta de Avelar2, In this study, the efficacy of this new pour- Márcia Cristina Alves Teixeira3, Dr. Marcio on formulation was demonstrated by the Dentello Lustoza3, Dr. José Manuel Moreno reduction of EPG counts greater than 80 Montroy4, Dr. Emmanuel Briant1, Dr. Christelle % until 28 days after treatment in Brazilian Navarro1, Dr. Vanessa Chala1 cattle naturally infested by gastro-intestinal 1Virbac, Carros, France, 2Universidade Federal nematodes. Rural do Rio de Janeiro, Seropédica, Brasil, 3Virbac do Brasil Industria e Comercio LTDA, PS03.03 Toxoplasma Gondii and Besnoitia São Paulo, Brasil, 4Laboratorios Virbac Mexico, Besnoiti Infections Differentially Modulate Guadalajara, Mexico Host Cell Cycle Progression in Primary Endothelial Host Cells Gastro-intestinal nematodes (Trichostrongylidae) are the most frequently Dr. Zahady Velasquez1, Dr. Ivan Conejeros1, Dr. observed parasites in grazing cattle worldwide Katharina Kerner2, Prof. Carlos Hermosilla1, causing digestive disorders, anemia, delays in Prof. Anja Taubert1 growth and production. The objective of this 1Insititute of Parasitology, Justus Liebig study was to test the efficacy of a new 0.5% University Giessen, Giessen, Germany, Eprinomectin, 5% Diflubenzuron-based pour- 2Institute of Hygiene and Infectious Diseases on formulation on the elimination of a natural of Animals, Justus Liebig University Giessen, gastro-intestinal nematodes infestation in a Giessen, Germany Brazilian cattle population. Background: Toxoplasma gondii was For this study, 20 male or female cross-breed described to modulate the host cellular cell animals B. indicus x B. taurus, 8 to 24 months cycle and to dampen host cell replication, old, 100-260 kg live weight, were selected while no respective data are available on from the experimental herd of Universidade Besnoitia besnoiti. However, T. gondii-related Federal Rural do Rio de Janeiro. Animals data are inconsistent since some authors were kept on pastures, without parasiticide detected a T. gondii-induced cell cycle arrest treatment and were naturally infested by in the G0/G1 phase whilst others report on a gastro-intestinal nematodes. On D0, these G2/M-based cell cycle arrest in infected cells. animals were randomized to the control Objective: We here analyzed effects of T. group (C, n=10) or the treatment group (T, gondii and B. besnoiti infections on host cell n=10) according to the average EPG counts cycle progression in primary endothelial cells. performed on D-7, D-2. On D0, 1 mL/10kg/

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 281 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Methods: Proliferation of infected and non- bovine polymorphonuclear neutrophils infected bovine umbilical vein endothelial (PMN). The purpose of this study was to cells was estimated microscopically. Cell analyze if pure B. besnoiti tachyzoite- or cycle phases (G0/G1-, G2/M-, S-phase) were calcium ionophore (A23187)-triggered analyzed by a FACS analysis of cellular DNA NETs would damage endothelial host cells content. Cell cycle-specific key regulatory and subsequently influence intracellular proteins (cyclin A2, cyclin B1 ) and histone H3 development and proliferation of B. besnoiti S10 as a marker of mitosis progression were tachyzoites in primary bovine endothelial analyzed via Immunoblotting. cells. Thus, we here evaluated endothelial host cell damage triggered by histone 2A Results: T. gondii-infections induced (H2A) and B. besnoitia tachyzoite-induced enhancement of host cellular proliferation NET preparations and furthermore estimated when compared to non-infected controls. In the effects of PMN floating over B. besnoitia- contrast, B. besnoiti infections had no effect infected endothelium under physiological on host cell division. Referring to their impact flow conditions on endothelial host cell on cell cycle phases, T. gondii induced a G2/M viability. Overall, all treatments (H2A, B. phase arrest in infected cells with impairment besnoiti-triggered NETs and floating PMN) of host cellular cytokinesis while B. besnoiti significantly induced endothelial cell death seemed to arrest cell cycle progression in of B. besnoiti-infected host cells. However, G0/G1 phase. B. besnoiti infections caused thought host cell damage led to significantly a decrease of cyclin A2 and of cyclin B1 altered intracellular parasite development with phosphorylation whilst in T. gondii-infected respect to meront diameter and numbers, BUVEC no respective alterations were the total proliferation of the parasite over found. In contrast to B. besnoiti, confocal time was not significantly affected by these microscopic analyses of the different treatments thereby denying any direct effect mitosis phases revealed that chromosome of NETs on intracellular B. besnoiti replication. segregation in T. gondii-infected cells was significantly impaired in infected cells, which occasionally presented more than two poles PS03.05 Strategic Control of Gastrointestinal of cell division. Thus, T. gondii impedes Nematodes in Beef Cattle in Brazil correct mitosis process and subsequent host cell division. Fernando Borges1, Dyego Borges1, Mariana Freitas1, Flávia Fávero1, Elvis Coelho Filho1, Conclusion: The current data show that T. Jonis Genro2, Welton Cabral3, Elio Moro3, gondii and B. besnoiti differentially interfere Frederico Moreira4 with cell cycle machinery of primary 1Ufms, Campo Grande, Brazil, 2Zoetis endothelial host cells and thus use different Animal Health, São Paulo, Brazil, 3Novapec, strategies of host cell modulation. Rondonópolis, Brazil, 4Zoetis Animal Health, Jersey City, United States

PS03.04 Bovine Neutrophil Extracellular The aim of this study was to compare Traps Induce Damage of Besnoitia Besnoiti- two protocols for strategic control of Infected Host Endothelial Cells but Fail to gastrointestinal nematodes in beef cattle Affect Total Parasite Proliferation in Brazil. Sixty Nelore weaned calves were distributed in three groups, based on the Dr. Ivan Conejeros1, Dr. Zahady D. Velasquez, number of fecal egg counts (FECs) and live Dr. Ershun Zhou, Prof. Anja Taubert, Prof. weight: T1 Placebo, T2 May – moxidectin Carlos Hermosilla 500 g/Kg, November – doramectin 700 g/ 1Justus Liebig University Giessen, Germany Kg, and T3 May – doramectin 700 g/Kg, Augustµ – moxidectin 200 g/Kg, Novemberμ Abstract: Besnoitia besnoiti tachyzoites – doramectin 700 g/Kg. Animals wereµ kept infect and develop in bovine endothelial in exclusive padocks with µan area repetition. cells in vivo and triggers the release of Individual weighingµ and faecal collections neutrophil extracellular traps (NETs) from were performed every 28 days from d0

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 282 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

(May/2017) to d313 (April/2018) after a 12 In the present study, we describe h fastening. At the end of the study, calves experimental infection in bull and heifer from T2 and T3 were 11.3 and 17.3 kg heavier calves with T. foetus trophozoites. We discuss than those of T1 (P<0.05) respectively, but culture and PCR findings following infection there was no difference between T2 and and describe the distribution of parasites in T3,. Two peaks of FECs were observed in host tissues. Our results provide a framework untreated animals: the first in May and June for future studies regarding diagnostic and and the second from October to December, treatment options for bovine trichomoniasis. with a predominance of Haemonchus sp. and Cooperia sp. The treatment with long-acting moxidectin resulted in a significant reduction PS03.07 Mini-FLOTAC Automated System of FEC in June (d 28) and maintained FEC reduced until September (d 112), whereas Alessandra Amadesi1, MP Maurelli1, Antonio this pattern was not observed in T3.. The Bosco1, Davide Ianniello1, ME Morgoglione1, treatment with a long acting formulation of Mirella Santaniello1, Laura Rinaldi1, Giuseppe moxidectin in T2 (May) had the same effect Cringoli1 on performance than using the first two 1Department of Veterinary Medicine and formulations in T3 (May and August). Both T2 Animal Production, University of Naples and T3 protocols can be used depending on Federico II, CREMOPAR, Napoli, Italy the level of resistance and the availability of handling the animals two or three times. In an era of technological revolutions in the diagnostic industries, also diagnostic methods for parasitic helminth infections are evolving, PS03.06 Development of a Natural Host for instance through the use of Mini-FLOTAC Model for Tritrichomonas Foetus techniques, the use of pooled samples, point- of-care diagnosis and automation of faecal Dr. Katy Martin1, Dr. Jeba R J Jesudoss egg count (FEC) and FEC reduction (FECR) Chelladurai1, Dr. Matt Brewer1 tests for assessing helminth infections and 1Iowa State University, Ames, United States anthelmintic resistance.

Tritrichomonas foetus is a sexually- The aim of this study was to design and test transmitted protozoan parasite and the cause a prototype of the Mini-FLOTAC automated of bovine trichomoniasis. Infection with T. system that allows a rapid laboratory workup foetus results primarily in early embryonic and can be used directly on livestock farms death, which in turn causes significant for automated FEC/FECR including image economic losses for producers. There are software analysis. currently no approved treatments for bovine trichomoniasis in the United States. Due to The scanning device for the Mini-FLOTAC was the economic impact of the disease, infected designed as an equivalent of a XYZ motorized animals are generally culled to prevent further microscopy stage that is automatically moved transmission of the disease. step by step to scan the two entire flotation chambers by taking partially overlapping Typically, only adult bulls maintain infection pictures. The movement of the stage is with T. foetus. This has contributed to controlled via software by a main processing significant challenges in the study of bovine unit, which can be either a PC, a Tablet trichomoniasis, due to the cost and potential or a Smartphone. The development of an safety concerns of maintaining bulls in image-analysis software is able to identify a research setting. We hypothesize that and count helminth eggs in order to reduce developing a calf infection model for the the time required for the analysis and the study of bovine trichomoniasis will allow for human errors. Furthermore, the development advancements in diagnostic tests, treatment of an automated software permits to options, and prevention strategies for the transmit collected data via internet to expert disease. diagnostic centers.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 283 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

The use on farm of this simple, automated side effects was 5%. Obstruction and choking system will allow a rapid assessment of FEC/ were not observed or reported during this FECRT in large and small ruminants to assist the study. Two serious adverse events (blindness new generation of veterinarians and farmers. and renal failure) were reported during this study and were most likely due to factors other than treatment administration (genetic PS03.08 Palatability and Safety of a Soft predisposition and long term exposure to Chewable Tablet Containing Ivermectin, non-steroidal anti-inflammatory drugs). Praziquantel and Pyrantel Pamoate in Client- Owned Adult Dogs The product was found to be well accepted (96.3% voluntary acceptance) and safe Christelle Navarro1, Frank Hurtig2, Sloane (low incidence of self-limiting side effects Joanes3, Dr. Vanessa Chala1 reported) during this trial. 1Virbac Global Medical, Carros, France, 2Virbac Corp., Fort Worth, United States, 3Virbac R&D, Fort Worth, United States PS03.09 Quantitative Proteomics Analysis of Angiostrongylus Vasorum-Induced Palatability and safety of heartworm Alterations in Dog Serum preventative treatments are crucial to ensure good compliance. Dr. Lucienne Tritten1, Dr. Tobias Kockmann Kockmann2, Dr. med. vet. Nina Gillis- An open-label field study was conducted at Germitsch1, Prof. Manuela Schnyder1 five veterinary clinics in Texas to determine 1Institute of Parasitology, University of Zurich, the palatability and safety of a new soft Zürich, Switzerland, 2Functional Genomics chewable tablet (Iverhart Max soft chew, Center ETH/UZH Zürich, Switzerland Virbac) under conditions of field use. There were 132 dogs enrolled in the study Blood contains hundreds of proteins, (including 14 dogs from breeds known to reflecting ongoing cellular processes bear the MDR1 mutation allele), who received and immune reactions. Infection with each 3 treatments at monthly intervals. The Angiostrongylus vasorum in dogs is number of doses taken voluntarily (offered associated with a perturbed blood protein by hand or in a bowl with or without food) profile, but existing reports lack the necessary was evaluated to determine the palatability depth of analysis to resolve the observed of the bacon-flavored chews. The reported pathologies in A. vasorum infections, adverse events were analysed to establish the including bleeding disorders. safety profile of the product. The incidence of vomiting, chocking, obstruction and Serum proteins from eight experimentally- other gastrointestinal incidents were closely infected dogs (i) before inoculation with A. evaluated to address any safety concerns vasorum, (ii) 34 days post-inoculation (p.i.; regarding dosing with the chew. immature infection), and (iii) 75 days p.i. (mature patent infection), were analyzed Three hundred and eighty-nine dosing events using liquid chromatography and tandem occurred during the study: 336 doses were mass spectrometry (LC-MS/MS). Sera from accepted as a treat (by hand or in empty two dogs were additionally examined at bowl) (86.3%), 39 in a bowl with food (10%) days 104 and 230 p.i.. A data-independent and 11 had to be pilled and placed in the acquisition workflow was employed in order mouth (3%). Three dogs (0.7%) could not be to generate quantitative data. Computational successfully dosed with this dosage form. analysis revealed 139 up- and down-regulated The adverse events reported by the owner proteins following infection (log2 ratio cutoff were diverse (vomiting (2.3%), diarrhea ≥ 1.0; q-value ≤ 0.05). or loose stool (1.2%), increased salivation (0.7%)), generally required little treatment Differences in serum profiles were most or follow-up and were self-limiting. Overall pronounced at day 75 p.i. when compared the incidence of potential gastrointestinal to before inoculation. Among up-regulated

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 284 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

proteins, chitinase 3, several saposin-like than adult dogs, while adult dogs were more proteins and heat shock proteins were greatly likely infected with T. vulpis (P<0.0001); egg/ increased (log2 fold-changes ≥ 5). Levels of cyst/oocyst per gram estimates for positive pulmonary surfactant protein B were already samples were not significantly different by elevated at day 34 p.i. in the prepatent phase. age class (P>0.05). Infection with T. vulpis Pathway enrichment analyses revealed that was significantly more common (P<0.0001) in complement (especially the lectin pathway) dogs also infected with A. caninum. Antigen and coagulation cascades as significantly testing for D. immitis was performed on 157 affected upon analysis of down-regulated dogs (≥5 months), 99 with fecal flotation, and proteins. Among them there were mannan- modified Knott’s test to detect microfilariae binding lectin serine peptidases, ficolin, and in blood. Heartworm antigen was detected coagulation factors. in 13 (8.3%) before and 16 (10.2%) after heat- treatment; microfilariae were detected in 9 These results reflect the ongoing immune (4.5%), including 7 dogs with D. immitis and response and stress imposed to the lungs 2 with Acanthocheilonema reconditum. Two by the parasite. In addition, they add new dogs with microfilariae of D. immitis were elements towards understanding the antigen negative before but positive after coagulopathies observed in A. vasorum- heat treatment of serum. Heartworm infection infected dogs. was not found to be significantly associated with age or detection of any parasites by fecal flotation (P>0.05). Intestinal parasites and PS03.10 Prevalence of Intestinal Parasites heartworm remain common in shelter dogs, and Heartworm in Northeastern Oklahoma with younger dogs particularly likely to be Shelter Dogs shedding protozoa and adult dogs more likely infected with T. vulpis. Kellee Sundstrom1, Megan Wohltjen1, Kelly Allen1, Susan Little1 1Department of Veterinary Pathobiology, PS03.11 Surveillance of Parascaris Equorum Oklahoma State University Center for Infections and the Efficacy of Fenbendazole Veterinary Health Sciences, Stillwater, United in Horses Across the United States States Dr. Brian Herrin1, Dr. Duane Chappell2, Dr. Intestinal parasites, including several zoonotic Bryant Craig2, Dr. Craig Barnett2, Dr. Earl agents, and heartworm (Dirofilaria immitis) Gaughanv, Dr. Wendy Vaala2 are commonly detected in shelter dogs 1Kansas State University, Manhattan, United due to inconsistent treatment and frequent States, 2Merck Animal Health, Summit, United exposure. To determine the prevalence of States intestinal parasites in fecal samples and evaluate interactions between age, sex, and Parascaris equorum is the most significant infection status, semi-quantitative centrifugal internal parasite pathogen of foals, presenting flotation in sugar solution (SG 1.25) was as respiratory disease as the juvenile performed on samples (4g) from 218 dogs ascarids migrate through the lung, and as with results evaluated by chi-square (P<0.05). gastrointestinal disease as the adult parasites Flotation revealed parasites in 173/218 (79.4%, remove nutrients or large populations cause 95% CI 73.5–84.2%) samples, including impaction colic or intussusception. The Ancylostoma caninum (118; 54.1%), Trichuris current study enrolled 410 horses on 27 farms vulpis (73; 33.5%), Toxocara canis (21; 9.6%), (197 weanlings; 213 horses >1 year old) from Cystoisospora spp. (43; 19.7%), Giardia sp. across the United States, to determine the (41; 18.8%), Cryptosporidium sp. (12; 5.5%), ascarid burdens by fecal egg count (FEC), and Sarcocystis sp. (4; 1.8%); 1 sample each as well as the efficacy of fenbendazole as (0.5%) contained Demodex sp., Dipylidium judged by fecal egg count reduction test caninum, or Taenia sp. Dogs <1 year of age (FECRT) from horses with any P. equorum were more likely to be shedding Giardia sp. eggs on the first FEC. Horse owners were (P<0.01) and Cystoisospora spp. (P=0.03) also asked to fill out a survey on their normal

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 285 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

deworming and management strategies. All method and the Modified McMaster horses under 1 year old received one dose of technique. Details of anthelminthic use were fenbendazole (10 mg/kg), and the horses > collected using a standardized face-to-face 1 year of age received 5 consecutive days of owner/horse-keeper questionnaire. The treatment of fenbendazole at 10 mg/kg. Of Student`s T-test of independence was used to the 130 horses that had an ascarid-positive investigate the association between exposure fecal, 107 of them were weanlings (p-value < variables and infection status/intensity. The 0.00001), and average ascarid FEC for those following helminthes were identified in the horses was 174 EPG (Range: 1 – 6,768). The study: Parascaris spp.(51.97%), Dictyocaulus overall efficacy of fenbendazole against P. spp.(17.32%), Strongylus spp.(14.96%), Oxyuris equorum was very high with a FECRT of 98.3% spp.(7.8%), Strongyloides spp.(2.36%), (Range: 83.0 – 100.0%). Because of the high Gastrodiscus spp.(3.15%), Anoplocephala efficacy, there were no significant differences spp.(0.79%), Habronema spp.(0.79%), and in egg reductions between age of animal, Trichonema spp.(0.79%). Of the 103 samples deworming strategy, or even between farms. examined, 87 (84.47%) were positive for Additionally, there were no management helminthes ova while 16 (15.53%) were practices that significantly changed the pre- negative. Parascaris equorum was the most treatment FEC or reduction of ascarids egg prevalent helminth among the sampled shedding. Infections with P. equorum continue horses, followed by Dictyocaulus arnfieldi and to be a significant problem for horses under then Strongylus spp. The class Nematoda has the age of one, but fenbendazole remains the highest percentage of infection (94.18%), a highly effective dewormer for controlling Trematoda (1.16%), Cestoda (1.16%) and mixed those infections. Overall, this study reiterates infections (3.48%). Eggs of Strongylides spp the need for a tailored deworming strategy were not seen. Intensities of helminth infection based on the horse’s age and other risk was found to decrease with horses age and factors, incorporating fecal egg counts into the use of proprietary equine anthelminthic program for assessing the deworming efficacy. products. There was no significant variation in the mean intensities of infection with respect to the location of stables as well as PS03.12 Coprological Prevalence and the sex of the animals sampled. In conclusion, Intensity of Helminth Infections In Polo and strongyle infection is endemic in the FCT but Work Horses (Equus caballus) In Abuja, equine anthelminthics assists in managing the Federal Capital Territory (FCT), Nigeria infection. Recommendations made include the need to institute wholistic parasite control Olorunfemi Jegede1, Victor Okorie1, Professor measures involving strategic deworming Maxwell Opara1, Sylvester Obeta1, Bala programme for stabled horses. Mohammed1 1University Of Abuja, Abuja, Nigeria PS03.13 Coproscopical, Antibody-Based This study aimed to estimate the prevalence and Molecular Survey on Equine Helminth and intensity of helminth infection of horses Infections on Horse Farms in Berlin/ (Equus caballus) in the Federal Capital Brandenburg, Germany Territory (FCT), Abuja, Nigeria and investigate the associations between infections and Laura Jürgenschellert1, Dr. Jürgen Krücken1, horses age, sex and owner-reported use of Dr. C.J. Austin2, Prof. Dr. Martin K. Nielsen3, anthelminthics. In a cross-sectional survey, Dr. Eric Bousquet4, Dr. Georg von Samson fresh faecal samples were obtained from Himmelstjerna1 103 horses in nine randomly selected horse 1Freie Universtität Berlin, Berlin, Germany, stables in three Area Councils of the FCT, 2Austin Davis Biologics Ltd., Great Addington, viz: Abuja Municipal Area Council (AMAC), United Kingdom, 3University of Kentucky, Gwagwalada and Kuje, and worm egg counts Lexington, United States, 4Virbac, Carros, performed at the parasitology laboratory, France University of Abuja using the direct faecal smear, floatation method, sedimentation Knowledge concerning the current prevalence

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 286 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

of helminth infections in horses is sparse, PS03.14 Anticoccidial Efficacy of Naringenin while at the same time this information is and a Grapefruit Peel Extract in Naturaly- urgently needed to guide new worm control Infected Growing Lambs with Eimeria Spp. approaches with less routine anthelmintic treatments. This study aimed at providing Master Agustin Pérez-Fonseca1, Dr. Yazmín representative data on the occurrence of Alcalá-Canto1, Dr. Cristina Guerrero-Molina1, the major groups of intestinal helminths on Dr. Aldo Alberti-Navarro1 horse farms in Berlin and Brandenburg. From 1Universidad Nacional Autónoma de México, a total of 484 horses on 48 farms faecal, Mexico City, Mexico saliva and serum samples were collected from May 2017 to January 2018. Faecal The current study aimed to determine samples were analysed using mini-FLOTAC the anti-Eimeria efficacy of an extract technique and combined sedimentation- of grapefruit peels (GF) and commercial flotation. Additionally, PCR on DNA from naringenin (NAR) in naturally-infected lambs, strongyle larvae was performed to investigate as well as the influence of these flavonoids the potential presence of Strongylus spp. on the oxidative status. Pharmacokinetic The latter was also approached by testing profiles were also determined. Extracts were serum samples for antibodies against administered per os to Eimeria naturally Strongylus vulgaris (rSvSXP-Antigen). Serum infected growing lambs during 90 consecutive (ADB IgG(T) serologic ELISA) and saliva days. The commercial anticoccidial drug (EquiSal® Tapeworm test) were examined toltrazuril (TTZ) was included in this trial as a for antibodies against Anoplocephala. standard. Twenty-four lambs were divided into In 66.9% of the faecal samples, eggs of four groups: NAR, lambs given a daily dose strongyles were found (64.2% in mini-FLOTAC of 5 mg of a commercial naringenin extract technique, 57.6% in combined sedimentation- of 98% higher purity per kg body weight; GF, flotation). Additionally, 1.2%, 0.6% and 0.4% lambs that recived a daily dose of 5 mg of of faecal samples were positive for Oxyuris ethanolic extract of grapefruit peels per kg equi, Anoplocephala spp. and Parascaris body weight; TTZ, lambs treated with 20 mg spp. eggs, respectively. Out of the 365 of toltrazuril/kg body weight on days 0 and 15 horses examined using the Anoplocephala of the experiment; and CTRL, untreated lambs saliva test, 29.6% were positive on 28 out that received daily dose of 30 ml of water. of 37 (75.7%) farms, while of 481 serum Daily doses of GF and NAR were dissolved in samples 16.2% were positive on 25 out of 30 ml of water. The CTRL group received 30 48 farms (52.1%). Regarding S. vulgaris, ml of water; as well as the TTZ group for the seropositivity was observed for 21.2% of period after the single dose administration. 481 samples corresponding to 83.3% farm Fecal and serum samples were collected from prevalence, while in 13 of the 484 examined all lambs. Anticoccidial efficacy was estimated faecal samples (2.7%) from 6 farms (12.5%) by coprological techniques. Generation Strongylus spp. DNA was detected. The of nitric oxide levels and the antioxidant unexpectedly high (sero-) prevalence data capacity of the experimental compounds encountered for both tapeworm and large were determined by the Griess and ABTS strongyle infections point at an urgent need assays, respectively. On day 30 post- for routine parasite surveillance. Due to its ingestion, anticoccidial efficacy was 91.76% high pathogenicity the exceptionally high (NAR) and 89.65% (GF); whereas 99.63% of seroprevalence of S. vulgaris is alarming and efficacy was achieved with TTZ 15 days after requires further attention, particularly in the treatment. NAR, GF and TTZ significantly context of ‘targeted selective treatment’. reduced oxidative stress in infected animals. Following the oral administration of NAR and GF, values in plasma approached maximum concentrations within 2.1 to 2.5 h. In conclusion, the administration of NAR and the GF extract reduced Eimeria oocyst output and oxidative stress in infected lambs.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 287 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.15 Mechanism and the Effects of PS03.16 Anthelminitc Efficacy and Urea on the Development and Hatching of Cytotoxicity of Diospyros Whyteana Trichostrongylus Colubriformis Eggs Compounds Derived Using Antifungal Activity

Mr. Robin McAnulty1, Mr. Nollan Puget2, Dr. Ms. Bellonah Sakong1, Prof. Mathew Adamu2, Andrew Greer1 Dr. Balungile Madikizela3, Dr. Maurice 1Lincoln University, Christchurch, New Awouafack4, Dr. Monnye Mabelebele1 Zealand, 2Agrosup Dijon, Dijon, France 1University of South Africa, Florida, South Africa, 2Federal University of Agriculture, Interrupting the parasite lifecycle outside Makurdi, Nigeria, 3University of Pretoria, of the host may provide novel methods Pretoria North, South Africa, 4University of of control that complement the use of Dschang, Dschang, Cameroon, 5University of anthelmintic drugs. Topical application of South Africa, Florida, South Africa Urea can interrupt nematode parasite egg hatching, although the mechanisms are still Gastrointestinal (GI) nematode infections are to be determined and are the objective of one of the major health challenges affecting this study. Firstly, the role of osmolality was performance of small stock. Furthermore, investigated using egg hatch assays with animals have developed resistance against either Urea, NaCl, Glucose or CaCl2 solutions the use of clinical drugs, and therefore, this at an osmolality ranging between 0 and led to the utilisation of alternative products. 1500 mOsmoles per l. All solutions reduced Hence, the aim of this study was to investigate percentage egg hatch from in excess of 85% if antifungal assays can be used as a model to in water to less than 3% at 1500 mOsmoles. isolate anthelmintic plant compounds, based However, there was a solution x concentration on potential correlations in the two activities. interaction whereby the lethal concentration Anthelmintic egg hatch assay (EHA) and (LC) 50 was 792 ± 32.1, 486 ± 41.2, 498 ± 36.5 the larval development assay methods were and 299 ± 36.8 mOsmoles per l for Urea, NaCl, used. Bioactivity of the fractions as well as glucose and CaCl22, respectively, being least cytotoxicity (IC50, tested using four human (P<0.05) in CaCl2, not different (P>0.05) cells in the colorimetric MTT assay) was between NaCl and Glucose (P>0.05) and assessed during fractionation, which resulted greatest in Urea (P<0.05). As the osmolality in isolation of three active compounds. of solutions increased the egg hatching The Diospyros whyteana acetone leaf extract percentage decreased and differences in egg had good activity with EC50 values of 4.43 hatching percentage between solutions at the and 5.58 g/ml respectively. The extract was same osmolality indicates that this effect is active against Candida albicans with MIC of not solely due to osmolality. The second study 40 g/ml,µ and this organism was used as a examined the penetration of Urea and NaCl model to isolate antifungal compounds from when applied to faeces. Following topical D. whyteana.µ EC50 values of the compounds application, faeces were left for 24h before ranged from 0.11 to 3.30 in the EHA and 1.5 samples measuring 1cm x 1cm were sectioned to 4.35 in the LDA, with promising selectivity throughout the faecal pat and placed in index (SI) values above 28. The three mini-bearman apparatus. The number of compounds had the same MIC of 160 ug/ larvae recovered from either NaCl or Urea ml against C. albicans. Conclusions: Results treated faeces were reduced relative to water showed good correlation between the EHA controls in the first 1 cm by 64% and by 34% and LDA as well as good antifungal activity, in the second 1 cm cube from the surface indicating that the use of antifungal assay only. Results indicate the mechanism of Urea assisted with the isolation of anthelmintic interrupting egg hatching is not solely due to compounds. Consequently, -amyrin and osmolality and when Urea is topically applied -amyrin were isolated and structures to faeces interrupts the egg development in elucidation were also identified.α the first 2cm of faeces nearest to the surface. β

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 288 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.17 Identification and Molecular LoVo cells in vitro. These findings provide Characterization of Exosome-Like Vesicles new insights into the interaction between Derived from Taenia Asiatica Adult Worms tapeworms and host mediated by exosome- like vesicles. Panhong Liang1, Li Mao1, Shaohua Zhang1, Xiaola Guo1, Guangxue Liu1, Lijie Wang1, Yadong Zheng1, Xuenong Luo1 PS03.18 Efficacy of a Permethrin and Fipronil 1Lanzhou Veterinary Research Institute, Combination (Effitix® Spot-On), Against Lanzhou, China Flea Infestations in Dogs: A Randomized, Controlled Study T. asiatica is one of the most important food-borne parasites and poses a great Dr. Fabio Barbour-Scott1, Dr. Thais Correia- threat to human beings. So far little is Azevedo1, Dr. Debora Azevedo Borges1, Dr. known about the specific infection and Difrey Ribeiro Campos1, Dr. Priscilla Cardim immune escape mechanisms of T. asiatica. de Oliveira1, Dr. Marcia Cristina Alves Teixeira2, Recently, exosome-like vesicles have been Dr. Marcio Lustoza2, Dr. Vanessa Chala3, Dr. emerging as a regulator in the interactions Stephane Bonneau4 between parasites and hosts, providing a 1Universidade Federal Rural do Rio de Janeiro, new direction for research on infection of T. Seropédica, Rio de Janeiro, Brasil, 2Virbac asiatica. Here, exosome-like vesicles were Brasil, R and D Department, São Paulo, Brasil, harvested from the excretory/secretory 3Virbac, Global Medical Department, Carros, products of cultured T. asiatica and isolated France, 4Virbac, R and D Department, Carros, by differential centrifugation. The purified France vesicles, ranging from 30 to 150 nm in size, were identified as exosome-like vesicles Flea infestations can seriously impair dog’s by transmission electron microscope and health through parasites they can transmit as nanoparticle tracking analysis. Proteomic Dipylium caninum, or allergic conditions they analysis showed that a total of 464 proteins can arouse (as Flea Bite Allergy)(1) or may were identified in the exosome-like vesicles worsen (as Atopy)(2). Thus an adapted flea (455 derived from T. asiatica and 9 derived control remains mandatory. from the human). Of these proteins, enzymes involved in metabolic processes, such as This study evaluates the efficacy of a glyceraldehyde 3 phosphate dehydrogenase, Permethrin (44.88%) plus Fipronil (6.01%) based fructose 1, 6 bisphosphate aldolase, cytosolic spot-on (Effitix®, Virbac, Brasil) against fleas malate dehydrogenase, enolase and so on, infestations, compared to a reference product were enriched. The abundant proteins from (Frontline ® top spot, fipronil 10 %, Boehringer, proteomic analysis, 14-3-3 and enolase, were Brasil) in adult dogs. Eighteen healthy Beagle present in the exosome-like vesicles as shown dogs were equally randomly allocated to group by immunogold labeling. In addition, we 1 (Frontline®), group2 (Effitix®) or control group found 20 known miRNAs present in T. asiatica (no treatment). Treatment was applied once on sRNA libraries. MiR-71 was the most abundant, D0, following manufacturer recommendations followed by let-7 and tas-miR-4989. Also, (at least 60mg/kg of permethrin and 6.7mg/ some other common miRNAs, such as miR- kg for Fipronil). Dogs were infested with +/- 1, miR-9 and miR-10 were also found in the 100 fleas on D-2, D5, D12, D19, D26, D33, D40, vesicles. We further validated the expression D47, and D54. Flea counts were performed of T. asiatica miRNAs, including 6 known accordingly on D2 and then 48 hours following miRNAs (tas-miR-71, tas-miR-1, tas-miR-7, other flea challenges. tas-miR-9, tas-miR-10, tas-let-7) and 3 novel miRNAs (tas-m0022-3p, tas-m0816-3p, tas- The mean geometrical efficacies against a m0082-5p). The nine miRNA abundance were preexisting infestation were very high as soon consistent with high-throughput sequencing as D2 (99.02% in group 1 vs 100 % in group 2), results. Additionally, we demonstrated that as well as the prevention of new infestations the exosome-like vesicles experimentally (100 % on D7, D21 and D28 for both groups, labeled with PKH67 were internalized by 99.76% vs 100% on D14, 98.45% vs 98.33%

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 289 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

on D35 and 95.42% vs 96.96% on D42 for mosquito feeding activities, as well as group 1 and 2 respectively, with no statistical variation of mosquito feeding between hosts. differences (p<0.05). Patterns of mosquito feeding such as points of insertion, probing, and digestion were This trial allows to confirm the rapid onset of also discernible from the recordings. From action of the permethrin-fipronil combination, this study, EPG could likely be implemented regarding a preexisting infestation, with no in future research to investigate the feeding living fleas as soon as 2 days after product patterns of other mosquito species and application. This spot-on offers beside a very pathogen acquisition and transmission. good protection against fleas over 6 weeks.

PS03.20 Can Cell Phones Detect Mosquitoes PS03.19 Using Electropetography (EPG) Infected with Canine Heartworm? to Surf the Waves of Host Choice in Aedes Aegypti Doyeon Park1, Haripriya Mukundurajan2, Joy Bowles3, Haluk Kalin1, Dr. Lindsay Starkey3, Dr. Victoria Pickens1, Dr. Astri Wayadande1 Byron Blagburn3, Dr. Manu Prakash2, Dr. Sarah 1Oklahoma State University, Stillwater, United Zohdy1,3 States 1School of Forestry and Wildlife Sciences, Auburn University, Auburn, United States, Electropenetography (EPG) was originally 2Department of Bioengineering, Stanford developed for use on Hemipteran phloem- University, Stanford, United States, 3College feeding insects to study feeding activities, of Veterinary Medicine, Auburn University, including points of pathogen acquisition and Auburn, United States transmission. Because mosquitoes feed in a similar manner, it is possible that EPG could Dirofilaria immitis is a filarial nematode and be used to study mosquito probing activities. the causative agent of canine heartworm. D. Host choice is often studied in mosquitoes immitis can be transmitted by mosquitoes by observing behavior when exposed to of the genera Aedes, Anopheles, Culex, a specific host characteristic. Using EPG, and Ochlerotatus. Within the mosquito, the host choice could be studied via direct nematode parasite matures through three measurements of mosquito probing activities developmental larval stages (L1, L2, L3). A while taking a blood meal from a given host. recent study has shown that mobile phones EPG recordings were taken of Aedes aegypti are capable of capturing acoustic data from females exposed to three different hosts. mosquito wingbeats. Since each mosquito The hosts included one typical male, one species has a different wingbeat frequency typical female, and one atypical male. The by which they attract mates, with just a typical hosts have a past of feeding mosquito brief recording (<1/10th of a second) these colonies and experiencing no issues with acoustic signatures can be analyzed to quickly regular feedings, including Aedes aegypti, and determine if mosquitoes belong to a species report average mosquito feeding in natural that is known to transmit different pathogens. environments. The atypical host reported We examined wingbeat signatures and flight below average mosquito feeding in natural duration patterns of D. immitis infected and environments and negative impact when non-infected Aedes aegypti to determine if feeding mosquito colonies: the mosquitoes wingbeat mobile phone recordings can be died shortly after the second feed. EPG used to distinguish infected mosquitoes from recordings from the three hosts were non-infected ones. Female mosquitoes were collected using a completely randomized recorded prior to and at various time points design. Present results show irregular feeding after feeding on infected or non-infected patterns in A. aegypti when exposed to the dog blood by placing individual mosquitoes atypical host, such as prolonged probing time into a chamber and recording for 60 lengths and disrupted probing and digestion. seconds using a standard mobile phone. To The results of this study demonstrate that uniformly analyze audio data, recordings were EPG provides quantifiable recordings of processed using an in-house Python script

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 290 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

to determine wingbeat frequency and flight known drug targets and antigens. Lastly, duration. One hundred sixty recordings were we’ve performed comparative analyses of the gathered, and mosquitoes were dissected transcriptomic state of the filarial ES system to confirm the presence and number of with the model nematode Caenorhabditis heartworm larvae. Our findings indicate that elegans to identify conserved transcripts and L3 (infective stage) infected mosquitoes have to elucidate differences between free-living significantly lower (mean=429 Hz) wingbeat and parasitic nematodes. We hypothesize that frequencies than age-matched negative cell-surface targets prioritized by this effort mosquitoes (mean= 577 Hz; p<0.0001). Flight are promising targets for the development of duration based on wingbeat recordings was new anthelmintics. substantially lower (17.6% of the time) in L3 infected mosquitoes than non-blood fed mosquitoes (35.9% of the time). We present PS03.22 Effects of Gastrointestinal data suggesting that wingbeat frequencies Nematode Parasitism on Growth and may be used to identify mosquitoes infected Reproductive Performance in Ewe Lambs in with D. immitis, a pathogen of veterinary and Ontario, Canada public health concern. Emma Borkowski1, Niel Karrow1, Paula Menzies1, Jacob Avula1, Brandon Lillie1, Andrew PS03.21 Mining the Excretory-Secretory Peregrine1 System of Filarial Nematodes for New Drug 1University Of Guelph, Guelph, Canada Targets Infection with gastrointestinal nematodes Dr. Paul Airs1, Nick Kemble1, Gabriella Muñoz1, (GINs) is an important cause of loss of Dr. Nic Wheeler1, Dr. Mostafa Zamanian1 productivity on sheep farms in Ontario. 1University Of Wisconsin-Madison, Madison, Efforts to quantify the effect of GIN infection United States on growth have demonstrated mixed results; investigation of its impact on reproductive Filarial nematode parasites significantly performance has been limited. This study impact human (e.g. Brugia malayi), veterinary evaluated the effect of GIN parasitism on (e.g. Dirofilaria immitis), and wildlife heath; growth and reproductive performance of but the fundamental mechanisms underlying ewe lambs under Ontario grazing conditions. parasitism are not fully understood. The Replacement Rideau cross ewe lambs (n excretory-secretory (ES) system of filarial = 140) on a farm in central Ontario were nematodes is likely the primary conduit followed for two years (2016-2017) from for the release of products essential to the nursing through their first lambing and establishment and maintenance of infection. lactation event. The lambs grazed from There has been significant progress in May to November of each year and were cataloging the complex makeup of filarial sampled every 6-8 weeks during both grazing ES products released at the host-parasite seasons and once at mid-gestation in March interface, and there is continued interest 2017. At each sampling the ewe lambs were in resolving the role of these products weighed, body condition scores assigned, (proteins, vesicles, and nucleic acids) in serum proteins and hematocrit measured, modulating host responses to infection. In and fecal egg counts (FECs) performed. contrast, comparatively little attention has Rainfall levels and numbers of infective larvae been paid to the structure and function of on pasture were unusually low during the the underlying ES system in filarial nematode first grazing season, but were more typical species. We have utilized low-input and of Ontario conditions in the second grazing spatial transcriptomic approaches to identify season. The three most common GIN species gene transcripts enriched in ES-associated were Haemonchus contortus, Teladorsagia cells and tissue. We have used these spatial circumcincta and Trichostrongylus spp. data to identify ES-enriched cell-surface Multivariate mixed models were generated receptors and to highlight transcripts with for weight change over time and litter size at spatial expression profiles that cluster with lambing. Gastrointestinal nematode FEC was

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 291 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

not a significant predictor of weight change were stratified by FEC, then balanced for in replacement ewe lambs during their first sex, weight, and genetic variation and fed two grazing seasons. However, higher FECs one of three levels of dietary CV (n=7 per at lambing were associated with larger litter group): 0 (control), 250, or 500g CV/day for sizes (p = 0.05), likely reflecting increased 10 weeks. The lambs were fed 1kg daily using periparturient egg rise in ewes with larger a combination of CV pellets and commercially litters. GIN-associated morbidity was not produced sheep pellets to achieve the observed during the study, indicating that targeted CV consumption. Weekly weight, infection levels may have been insufficient to FEC, and packed cell volume (PCV) were negatively impact productivity. Therefore, determined. The FEC of lambs fed 500g CV subclinical GIN infection appears to have were significantly lower than those of control minimal impact on growth and reproductive lambs at week 1 (p = 0.02, 864 ± 205 vs 1807 performance in Ontario sheep. ± 387; mean ± SEM) and week 2 (p = 0.03, 800 ± 273 vs 1671 ± 289). The FEC decreased in each group over the duration of the trial. PS03.23 Anthelmintic Efficacy of There was no effect on PCV or weight gain. Nutritionally Balanced Cranberry Vine Pellet Further research is needed to establish Against Strongyle Infection in Dorset Lambs the optimal time to feed CV for maximum anthelmintic effect against GIN. Bailey Chalut1, Marissa Brummett1, Dr. Carly Barone2, Dr. Anne Zajac3, Dr. Michael Thonney4, Dr. Jess Reed5, Dr. Christian PS03.24 Chromosome-Scale Assembly Krueger5, Dr. Katherine Petersson1 of Haemonchus Contortus, a Model 1University Of Rhode Island, Kingston, United Gastrointestinal Parasite States, 2Bia Diagnostics, Colchester, United States, 3Virginia Tech, Blacksburg, United Dr. Stephen Doyle1, Dr. Roz Laing3, Mr. States, 4Cornell University, Ithaca, United Alan Tracey1, Dr. David Bartley3, Dr. Alison States, 5University of Wisconsin, Madison, Morrison3, Dr. Kirsty Maitland2, Dr. Nancy United States Holroyd1, Dr. Matthew Berriman1, Dr. Neil Sargison4, Dr. Eileen Devaney2, Dr. John Plants containing secondary compounds Gilleard5, Dr. James Cotton1 such as proanthocyanidins have been shown 1Wellcome Sanger Institute, Hinxton, United to suppress gastrointestinal nematode (GIN) Kingdom, 2University of Glasgow, Glasgow, infections, providing promise for alternative United Kingdom, 3Moredun Research institute, methods of GIN control in small ruminants. Penicuik, United Kingdom, 4University of To date, in vivo studies from our laboratory Edinburgh, Edinburgh, United Kingdom, have demonstrated a decrease in fecal egg 5University of Calgary, Calgary, Canada count (FEC) after six weeks of feeding cranberry vine (CV), however palatability Haemonchus contortus is an economically and nutritional quality limited consumption important gastrointestinal parasite of small of the CV. The purpose of this study was ruminants and a model for anthelmintic to test the anthelmintic efficacy of feeding research. Building upon the draft genome varying amounts of CV incorporated into published in 2013, we have completed and nutritionally equivalent to a 16% sheep a chromosomal-scale assembly for the pellet on lambs naturally and experimentally MHco3(ISE) isolate, which now represents infected with GIN. Cranberry vine prunings the largest, most contiguous assembly of any were incorporated into a feed pellet parasitic nematode to date. We describe the containing 500g CV in 1kg. Five-month-old distribution of genetic diversity throughout parasite naive lambs were turned onto pasture the genome from within and between lightly infected with GIN. An experimental globally distributed parasite populations, infection (5000 H. contortus L3) was and together with a significantly improved superimposed on the natural infection after transcriptome annotation aided by full-length one week on pasture to ensure adequate cDNA sequencing, we explore coordinated infection. After the infection matured, lambs gene expression throughout the life cycle.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 292 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

The assembly and annotation is available C. elegans N2 strain. We show that activation on WormBase Parasite (https://parasite. of parasite receptors in body wall and wormbase.org/Haemonchus_contortus_ pharyngeal tissue can be measured in simple prjeb506/Info/Index/), providing a significant plate-based assays and through the use of genomic resource for a broad group of Clade electropharyngeogram (EPG) recording, V nematodes, which include parasitic species and that activation of parasite receptors in of major veterinary and medical importance. the ASH sensory neuron can be measured This removes some of the dependency on through microfluidic trapping and single- using C. elegans as a genome reference for neuron calcium recordings. The suitability which parasite-specific traits are not relevant. of these approaches for a given parasite This chromosome-scale assembly now allows GPCR will depend on the complement of insight into chromosome evolution among related receptors and endogenous ligands these species, and offers a robust scaffold for that signal in targeted tissues. We show that genome-wide analyses of important parasite transgenic strains can be created in various traits such as anthelmintic resistance. genetic knockout backgrounds to help mitigate these concerns. While expression in scalable single-cell systems will remain PS03.25 Tissue-Specific Heterologous an important objective for high-throughput Expression of Filarial Parasite G screening (HTS) against GPCR targets, Protein-Coupled Receptors (GPCRs) in functional parasite receptor assays in a Caenorhabditis Elegans more native nematode cell and physiological environment can provide important baseline Kendra Gallo1, Dr. Nicolas Wheeler1, Tran To1, pharmacological data. Further, it is possible Eric Chen1, Athena Golfinos1, Cole Gilsdorf1, that these transgenic whole-organism assays Troy Meikle1, Dr. Mostafa Zamanian1 can be ultimately adapted for medium or 1University of Wisconsin Madison, United high-throughput screening. States

Parasite G protein-coupled receptors PS03.26 A One Health Approach: Translating (GPCRs) remain unexploited as anthelmintic Livestock Parasite Research to Scan for targets despite their involvement in critical Benzimidazole Resistance Mutations in sensory, neuromuscular, and physiological Human Soil Transmitted Helminths processes. One significant bottleneck in exploring the pharmacology of parasite Ms. Abhinaya Venkatesan1, Dr. Russell GPCRs results from difficulties in consistently Avramenko1, Dr. James Wasmuth1, Dr. Matt establishing heterologous expression in Workentine1, Dr. Piet Cools2, Dr. Bruno single-cell systems. Yeast and mammalian Levecke2, Prof. John Gilleard1 cell culture systems have paved the way 1Faculty of Veterinary Medicine, University for deorphanization of helminth GPCRs, of Calgary, Calgary, Canada, 2Faculty of but not all receptors express or behave Veterinary Medicine, University of Ghent, properly in cell types derived from distant Ghent, Belgium phylogenetic lineages. The combinations of accessory proteins, molecular chaperones, Soil Transmitted Helminth (STH) infections G proteins, and membrane determinants are a major health concern among developing required for the successful folding, cell- nations. Mass Drug Administration (MDA) surface expression, and signaling of parasite programs that aim to reduce the morbidity receptors in surrogate systems have not been associated with STH infections involve the comprehensively identified. To avoid some of administration of benzimidazole drugs to these complications, we’ve established new school age children considered to be at risk of endpoints for parasite GPCR expression in the infection in endemic regions. The increasing model nematode C. elegans. Building upon use of these drugs leads to a greater risk the work of others, we have expressed Brugia of the emergence of drug resistance which malayi aminergic receptors in the body wall would potentially confound the aims of the muscle, pharynx, and sensory neurons of the MDA programs. However, the detection

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 293 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

of resistance in STH the field is extremely no reliable in vitro biochemical assays to challenging and unreliable. Benzimidazole study pharmacological responses in the resistance is common and well characterized L3 or microfilarial stages. The currently in parasitic nematodes of livestock and used resistance validation methods include several Single Nucleotide Polymorphisms experimental infection establishment in dogs (SNPs) in the -tubulin genes encoding the and microsatellite analyses, both of which drug targets are known to be associated are expensive and time-consuming. We with resistance.β We have developed a report the evaluation of in vitro biochemical number of deep amplicon sequencing assays to test responses to drugs in validated assays, using the Illumina Miseq platform, resistant and susceptible isolates of Dirofilaria to undertake large scale scanning for these immitis microfilariae. These assays test benzimidazole resistance associated SNPs response parameters of microfilaria such as in gastro-intestinal parasites of ruminants. cell membrane integrity, cellular metabolism, We are now applying the same approach apoptosis, and efflux proteins to dilutions to investigate the potential emergence of of FDA approved MLs. We discuss how the benzimidazole resistance in the major human presented data can supplement the currently soil transmitted helminths (STH); Necator used resistance validation methods to identify americanus, Ancylostoma duodenale, Trichuris isolates of drug resistant heartworm. trichiuria and Ascaris lumbricoides. We have combined phylogenetic and RNAseq analysis of the complete -tubulin gene families in PS03.28 Effects of Alpha-Terpinene, each of the major STH species to prioritize Citronellal, Citronellol and Limoene in the genes for targeting.β Primer sets have been Caenorhabditis Elegans Motility Test designed to amplify fragments encompassing the relevant codons - 167, 198 and 200 - of Dr. Luciana Katiki1, Jessica Carolina Ribeiro1, our prioritized genes and the assays are deep Bruna C. Rodrigues Wedy1, Danielle Rizatto amplicon sequencing assays currently being Balbino1, Guilherme Fávero Luciani1, Poliana optimized. Once the assays are optimized and Araújo Pacheco2, Rodrigo Giglioti1, Cecília validated, we they will be used to screen STH José Veríssimo1, Helder Louvandini2, Karen populations from endemic regions, including Tiemi Akashi3 Tanzania and Ethiopia, to detect and quantify 1Institute of Animal Science (IZ/SAA), Nova the presence of benzimidazole resistance Odessa, Brazil, 2University of Sao Paulo, associated SNPs. Piracicaba, Brazil, 3State University of Maringa, Umuarama, Brazil

PS03.27 Evaluation of Biochemical Assays Caenorhabditis elegans is a small nematode for the In Vitro Testing of Drug Response in that can be maintained at low cost and the Canine Heartworm Dirofilaria Immitis handled using standard in vitro techniques. Microfilaria Unlike other in vitro tests, C. elegans motility assays provide data from a whole animal Dr. Jeba R J Jesudoss Chelladurai1, Krystal with intact and metabolically active systems. Magiera1, Dr. Katherine Martin1, Dr. Matthew T The model has the potential to inform Brewer1 farmacological doses, as well as in vivo studies 1College Of Veterinary Medicine Iowa State and also to predict active doses. Plant and our University, Ames, United States derivates including essential oils are globally acclaimed for their medicinal and therapeutic Dirofilariosis caused by Dirofilaria immitis values, especially anthelmintic effect. The is a source of great suffering in dogs and present study was designed to explore the cats in endemic regions in the United effect of different concentrations (5; 2.5; 1.25; States. Resistance development to the 0.6 mg/ml) of alpha-terpinene, citronellal, prophylactic macrocyclic lactones (MLs) is citronellol and limonene on C. elegans motility emerging in these regions and is a cause for test. For each concentration, 6 replicates were concern. Although bioassays and molecular performed diluted in Tween and negative markers have been developed, there are control with Tween and distilled water. Data

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 294 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

from the motility were analyzed by the third-stage larvae isolated by peptic digestion PROC GLM procedure of SAS, which model of infected Cornu aspersum snails (5 cats) or included fixed effects of oil, concentration and with tissues of an infected snail (1 cat). Snails interactions. The means between different had been inoculated with T. brevior isolated oils and doses were compared by the Tukey’s from a cat from Sicily. Starting 19 days post test (P <0.05). The means of the highest inoculation (dpi) fecal samples of 10 grams concentrations (2.5 and 5 mg/mL) differed each were collected daily and subjected to the from the control. Citronellol at the lowest Baemann technique. At 38 dpi, two cats were concentration had 100% mortality and must necropsied for lungworm recovery and count. be evaluated in lower doses to calculate LC50. The LC50 followed by their confidence limits None of the cats showed any respiratory signs for citronellal were 2.48 mg/mL (2.18±285), during the observation period. Cats started 4.16 mg/mL (3.44±5.50) for limonene and 5.11 shedding larvae at 21 dpi and all cats shed mg/mL (3.20±16.39) for the alpha-terpinene. larvae from 23 dpi onwards. Arithmetic mean Results suggest that the anthelmintic effect fecal larva counts peaked at approximately 40 of the oils on nematode C. elegans and dpi (831 larvae per gram feces). Necropsy of the potential to be investigated on other two cats revealed 16 (cat inoculated with 16 nematodes. - FAPESP 2017/23540-1. larvae) or 112 (cat inoculated with snail tissue) adult T. brevior. Larval shedding from the four cats followed up ceased 107 dpi, 141 dpi, 141 PS03.29 New Insights in the Biology of dpi or 227 dpi for the cats inoculated with Troglostrongylus Brevior 53, 12, 29 and 13 third-stage T. brevior larvae, respectively. Martin Knaus1, Emanuele Brianti2, Domenico Otranto3, Steffen Rehbein1 1Boehringer Ingelheim Vetmedica GmbH, PS03.30 Development of an Automated Kathrinenhof Research Center, Rohrdorf, Motility Assay Using Infrared Tracking Germany, 2Dipartimento di Scienze on Exsheathed Third- Stage Larvae Veterinarie, Università degli Studi di Messina, of Haemonchus Contortus for New Messina, Italy, 3Dipartimento di Medicina Anthelmintics Discovery Veterinaria, Università degli Studi di Bari “Aldo Moro”, Bari, Italy Dr. Beatriz Munguía1, Dr. Jenny Saldaña1, PhD Inés Carrera1, Manuela Ferrer1, Ramiro Teixeira1, While feline lungworms share a similar Elisa Melian1, Dr Laura Domínguez1 life-cycle with terrestrial gastropods as 1Laboratory of Pharmacology, Faculty of intermediate hosts, other aspects of their Chemistry, Universidad De La República, biology may differ. First contributions to the Montevideo, Uruguay biological characteristics of Troglostrongylus brevior were reported by Gerichter in 1949 Haemonchus contortus is one the most who verified the infectivity of T. brevior prevalent nematodes in gastro-intestinal third-stage larvae, after developing in snails, parasite infections of small ruminants and for the domestic cat. More recent research responsible of major economic losses in focused on the understanding of the biology livestock production. Several anthelmintic of the larvae within the intermediate host, drugs are available in the market to control potential pathways of transmission and risk this helminthiasis, but development of factors for transmission. However, several resistance against all major classes, has been questions concerning the development of T. reported. Therefore, the development of brevior in the final host remain open, including novel anthelmintic compounds in veterinary the duration of larval shedding and their medicine is essential. Availability of parasitic variability in individual definitive hosts. stages for in vitro screening relies on artificial infections and adult stages require ruminant In preparation for a clinical study, six domestic necropsy. Contrary, exsheathed L3 (xL3) short hair cats (8 to 9 months old, 2 male, 4 parasitic stage, can be easily obtained starting female) were inoculated orally with 12 to 53 from eggs in feces. In this work we show the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 295 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

set-up of a whole-organism automated motility Research Institute, Pentlands Science Park, assay for new anthelmintics discovery on United Kingdom, 8UMR 1225 IHAP INRA/ xL3 stage of H.contortus (Kirby anthelmintic- ENVT, France susceptible McMaster isolate). xL3s were cultured in multiwell plates according to Helminth parasitic pathogens cause severe Preston et al 2015, with minor modifications disease and are amongst the most important and the larval motility was measured for 30 production-limiting diseases of grazing min at 37°C, 72 hs post-incubation using an ruminants. Frequent anthelmintic use to infrared tracking device with temperature control these infections has resulted in control (WMicrotracker Arena, PhylumTech). the selection of drug resistant helminth This system uses video recording of infrared populations. Anthelmintic resistance (AR) microbeams illumination through multiwell is today found in numerous major helminth plates and the analog signal is mathematically species across the EU and globally. COMBAR processed to detect the passage of the larvae (COMBatting Anthelmintic Resistance in in the sensing area. This assay was calibrated Ruminants) is a COST Action (launched in using albendazole, monepantel and ivermectin 2017) which aims to advance research on as standards, with different mechanism of the prevention of AR in helminth parasites of action, and the half-maximum inhibitory ruminants and disseminate current knowledge concentration (IC50) was determined for each among all relevant stakeholders. COMBAR anthelmintic from four independent assays aims to integrate, evaluate and assess the with six replicates of each concentration economic trade-off of the novel developments evaluated, being 0.81±0.23, 0.10±0.04 and in the field mainly by networking and has 0.16±0.04 M (average ± standard deviation) already attracted scientists from 31 countries. respectively. Also controls of culture medium The Network has been organised around and cultureµ medium with DMSO were also three Working Groups (WG): WG1 “Improving carried out. Due to its fully automated Diagnosis” which aims to prioritise, evaluate characteristics for reading this is a rapid and implement cost-effective methods for the method for the screening of new compounds diagnosis of helminth infections and AR; WG2 with potential anthelmintic activity using a “Understanding the socio-economic aspects” easily obtained parasitic stage of H.contortus. which aims to develop, disseminate and apply methods to study the economics and human Acknowledgements: ANII (Uruguay), behaviour in the field of helminth control in PEDECIBA Química (Uruguay). ruminants and WG3 “Innovative, sustainable References: Preston, S. et al. Int. J. Parasitol., control methods” which aims to develop 2015, 45, 333-343. practical and sustainable helminth control strategies that integrate current insights from diagnostics, Targeted (Selective) Treatment PS03.31 Can We Combat Anthelmintic approaches, epidemiology, anti-parasitic Resistance in Ruminants? forages, vaccinology, farm economics and human behaviour. Dr. Johannes Charlier1, Dr. Smaragda Sotiraki2, Prof Laura Rinaldi3, Prof Edwin Claerebout4, Prof Eric Morgan5, Prof Georg von Samson- PS03.32 A Putative Cryptic Freeze-resistant Himmelstjerna6, Dr. Dave Bartley7, Dr. Herve Species of Trichinella Discovered in Wolverine Hoste8 1KREAVET, Belgium, 2Veterinary Research Peter Thompson, Rajnish Sharma2, Eric Institute HAO- DEMETER, Greece, 3University Hoberg3, Brad Scandrett4, Kelly Konecsni4, of Naples Federico II, Italy, 4Laboratory of N Jane Harms5, Piia M Kukka5, Tom S Jung5, Parasitology, Faculty of Veterinary Medicine, Brett Elkin6, Robert Mulders6, N.C. Larter7, Ghent University, Belgium, 5Queen’s University Marsha Branigan8, Jodie Pongracz8, Brent Belfast, Institute for Global Food Security, Wagner2, Benjamin Rosenthal1, Emily Jenkins2 United Kingdom, 6Freie Universität Berlin, 1USDA, Agricultural Research Service, Animal Institute for Parasitology and Tropical Parasitic Diseases Laboratory, Beltsville Veterinary Medicine, Germany, 7Moredun Agricultural Research Center, United States,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 296 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

2Department of Veterinary Microbiology, wolverine in northwestern Canada, especially Western College of Veterinary Medicine, Yukon, suggesting a possible historical link University of Saskatchewan, Canada, 3Museum to Beringia and the Palearctic. Exploration of of Southwestern Biology and Department of Alaskan and Siberian isolates may contribute Biology, University of New Mexico, United to further resolution of a geographically States, 4Centre for Food-borne and Animal complex history for Trichinella and other Parasitology, Canadian Food Inspection Agency, parasites across the Western Hemisphere, Saskatoon Laboratory, Canada, 5Department Beringia and Eurasia. of Environment, Yukon Government, Canada, 6Environment and Natural Resources, Government of the Northwest Territories, PS03.33 The New Rat Lungworms: The Canada, 7Government of the Northwest Occurrence of Physaloptera Hispida and a Territories, Department of Environment and Mastophorus Sp. In Pulmonary Vessels of the Natural Resources, Canada, 8Environment Hispid Cotton Rat (Sigmodon Hispidus) From and Natural Resources, Government of the Georgia, U.S.A. Northwest Territories, Canada Mr. Alec Thompson1, Dr. Christopher Trichinellosis is caused by nematodes of Cleveland1, Mr. Troy Koser1,2, Mr. Seth the genus Trichinella, which has two major Wyckoff1,2, Dr. Michael Yabsley1,2 clades. The encapsulated clade includes 1Southeastern Cooperative Wildlife Disease six species and three currently unnamed Study / University of Georgia, Athens, United genotypes, whereas the non-encapsulated States, 2Warnell School of Forestry and clade includes three species. In North Natural Resources, University of Georgia, America including Canada, five species of Athens, United States Trichinella (T. spiralis (T1), T. nativa (T2), T. pseudospiralis (T4), T. murrelli (T5) and During 2017-2018, a survey for the rat Trichinella genotype T6) have been found lungworm, Angiostrongylus cantonensis in homeothermic vertebrate hosts. Here we (Nematoda: Metastrongyloidea) in rodents discuss the discovery, phylogeny, geography from Piedmont and Lower Coastal Plains and host range of a previously unrecognized, physiographic regions of Georgia, U.S.A. was encapsulated, cryptic species of Trichinella conducted. On four occasions, a single worm designated as the T13 genotype. This novel was recovered from the pulmonary vessels genotype was discovered while sequencing of a single cotton rat (Sigmodon hispidis). the mitochondrial genome of an isolate One of these worms was identified as a mistakenly determined to be T. nativa based Physaloptera sp. and the remaining three as a on multiplex PCR. Subsequent phylogenetic Mastophorus sp. by morphology. Physaloptera analysis showed that the new genotype is (Nematoda: Physalopteroidea) and at the base of the subclade containing T. Mastophorus species (Nematoda: Spiruroidea) patagoniensis (T12), T. nativa, T. britovi (T3), are stomach parasites of many wild and T. murrelli (T5), and genotypes T6, T8, and domestic animals. This is the first report of T9 which is sister to the lineage containing T. these species in the pulmonary vessels of a spiralis (T1) and T. nelsoni (T7). Of 95 animals definitive host. To better characterize these from Canada that tested positive for T. parasites, representative specimens were nativa based on multiplex PCR, 14 wolverines collected from cotton rat stomachs and (Gulo gulo) were infected with the novel identified morphologically and molecularly. genotype. These occurred mostly as single Based on partial cytochrome c oxidase infections (11/14), but occasionally as mixed subunit 1 (COI) gene sequences, Physaloptera infections with T. nativa and Trichinella T6. hispida from stomachs were identical to We recommend use of a newly developed the Physaloptera sp. from the pulmonary PCR-RFLP or DNA sequencing to confirm vessels. The COI sequences from the identification of the isolates of T. nativa or Mastophorus sp. from the stomach exhibited any species of Trichinella, respectively. Our a higher degree of variability but confirmed results indicate that T13 is not geographically that the pulmonary worms were the same widespread in Canada, and may be limited to Mastophorus species. Furthermore, sequences

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 297 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

of Mastophorus from a coastal site clustered shock rather than temperature itself is a separately from a clade of Mastophorus crucial aspect in triggering the biological sequences from cotton rats from a Piedmont exsheathment cascade, and thus infection site. Collectively, our data show that adult process, of H. contortus. worms recovered from pulmonary vessels of cotton rats could be either Physaloptera or Mastophorus sp., indicating that these PS03.35 Conserved and Divergent Aspects parasitic worms are not always restricted to of Chemosensory Signaling in Filarial the stomach and that worms from pulmonary Parasitic Nematodes vessels must be carefully examined to obtain a definitive diagnosis of A. cantonensis infection. Dr. Nic Wheeler1, Zachary Heimark1, Tran To 1, Eric Chen1, Dr. Paul Airs1, Dr. Mostafa Zamanian1 PS03.34 Nematode Exsheathment 1: Heat 1University Of Wisconsin-Madison, Madison, Shock and Haemonchus Contortus United States

Dr. Tania Waghorn1, Dr. Kiliana Bekelaar1, Dr. Chemosensation - the process of receiving Mike Tavendale1, Catherine McKenzie1, Dr. Dave exogenous chemical cues, initiating a Leathwick1 signaling pathway, and precipitating 1Agresearch, Palmerston North, New Zealand an evolved behavioral response - is a fundamental life process that is shared from Understanding the processes involved in the animals to bacteria. The model nematode transition from the free living to the parasitic Caenorhabditis elegans has been shown to life stage of ruminant parasitic nematodes display positive and negative chemotaxis to may possibly identify new targets amenable soluble and volatile compounds, and much to future intervention. The transition to is known regarding the neuronal networks parasitism is initiated by exsheathment and molecular pathways underpinning these and is triggered by the sudden change in behaviors. Some of these pathways have been environment after ingestion of the infective recently explored in parasitic nematodes, larva by the host. Two major changes in particularly soil-transmitted helminths. In environment are the increases in temperature contrast, there is very little known about and carbon dioxide (CO2) levels. For CO2 chemosensory behaviors in vector-borne a role in exsheathment has been described filarial parasites such as Brugia malayi and previously, but the exact role of temperature Dirofilaria immitis, the causative agents of was unclear. This study investigated the the neglected tropical disease lymphatic importance of temperature in triggering filariasis and dog heartworm, respectively. exsheathment of Haemonchus contortus. We hypothesized that chemosensory Carbon dioxide induced exsheathment in signaling is integral to the migration and H. contortus proved to be temperature transmission of filarial parasitic nematodes, dependent, as no exsheathment was observed and that sensory pathways could present at room temperatures. The temperature new targets for vector or chemotherapeutic requirement to trigger exsheathment methods of filariasis control, and we have was quite specific, with a rapid change in taken the first steps toward elucidating the temperature (heat shock) very efficiently molecular mediators of chemotaxis in filarial inducing high levels of exsheathment. In nematode species. Pan-phylum analyses of contrast, when the larvae were exposed the nematode chemoreceptor GPCR family, to a slow increase in temperature the as well as TRP and CNG channels that are exsheathment response was smaller and hypothesized to function downstream of delayed. Further investigation revealed that chemoreceptor activation in the amphid timing of the heat shock in relation to the sensory neurons, revealed that filarial CO2 administration was crucial, as well as the nematodes have a reduced and divergent final temperature and magnitude of the heat subset of chemoreceptors, while they retain shock. one-to-one homologs to the canonical TRP In conclusion, these data indicate that heat and CNG families annotated in C. elegans. We

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 298 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

show enrichment of expected chemosensory erythrocyte count (TEC), haemoglobin proteins in the parasite anterior and explore and packed cell volume (PCV) and an stage-specific and temperature-dependent increase in total leucocyte count (TLC) and aspects of chemosensory receptor expression. eosinophilia. The biochemical studies revealed Using plate-based chemotaxis assays, hypoalbuminemia and hyperglobulinemia. we show that in vitro chemotaxis can be dysregulated by pharmacological and reverse genetic approaches targeting chemosensory PS03.37 Metazoan Parasitic Infections for proteins. Lastly, using long-read isoform Clarias Gariepinus (Burchell, 1822) in Native sequencing, we have cloned B. malayi and Extralimital Distribution Ranges Within osm-9 and tax-4 and are currently testing Southern Africa transgenic rescue of chemosensory function in corresponding C. elegans knockout strains. Miss Marliese Truter1,2,4, Dr. Kerry A Hadfield1, Prof. Olaf LF Weyl2,3,4, Prof. Nico J Smit1 1Water Research Group, Unit for PS03.36 Clinico- Histopathological Environmental Sciences and Management, Observations of Camel Sarcopticosis: A Case North-West University, Potchefstroom,, South Report Africa, 2DST/NRF Research Chair in Inland Fisheries and Freshwater Ecology, South Dr. Abhilasha Dadhich1, Dr. Manish Agarwal1, African Institute for Aquatic Biodiversity, Dr. Manisha Mathur1 Grahamstown, South Africa, 3South African 1Rajasthan University of Veterinary and Animal Institute for Aquatic Biodiversity (SAIAB), Sciences, India, Bikaner, India Grahamstown,, South Africa, 4Centre for Invasion Biology, Grahamstown, South Africa Cutaneous ectoparasitoses is one of the important skin diseases of camels and As one of the most widespread freshwater ranks second commonest infestation after fishes and an economically important fish in trypanosomosis. Sarcopticosis affects Africa, few studies investigate the parasitic productivity but mostly cause debilitation communities of the African sharptooth leading to heavy economic loss. Camel catfish, Clarias gariepinus (Burchell, 1822) in sarcopticosis also known as sarcoptic mange southern African freshwater systems. More is common at all ages in camels. It is usually specifically, studies investigating the parasitic a chronic condition with high morbidity communities and what determines the and low mortality. In this study, a skin composition of these parasitic communities sample was collected from camel (Camelus in both the native and extralimital distribution dromedarius) suspected for cutaneous range are limited. A recent review emphasises lesions, was processed mechanically the potential for C. gariepinus to be a for paraffin embedding by acetone and suitable host for various parasite taxa, with benzene technique for histopathological ca. 106 parasitic species across seven major examination. The blood sample was also metazoan taxa recorded from C. gariepinus collected from suspected case for analysis in Africa. Currently only ~24 parasite species of various haemato-biochemical parameters. are known from C. gariepinus in South Sarcopticosis was confirmed after the mites Africa. In addition, in recent years only five were identified as Sarcoptes scabei var descriptions of new species from South Africa cameli from the skin scrapings. The affected highlighting the paucity of knowledge on areas in head and neck showed papules and the parasitic communities of C. gariepinus eruptions. Microscopically, minute cavities in southern Africa. The present study aims in the epidermal layer extending into dermis to investigate the metazoan parasitic were present. These cavities were filled with communities of C. gariepinus in native and tissue debris and infiltrated mononuclears. extralimital distribution ranges in southern The epidermis showed hyperkeratosis and Africa. Furthermore, it aims to determine acanthosis and there was proliferation of which invasion mechanisms (i.e. parasite fibrous connective tissue. Haematological spillback or enemy release) are at play and observations revealed decrease in total how the diversity and abundance of these

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 299 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

parasite communities change throughout the studies to assess male/female ratio of flies distribution range, taking into account the age collected around the eyes of human baits, the of the fish, metal concentrations and nutrient T. callipaeda natural infection rate of flies and levels in the environment as well as the method the potential overwintering of T. callipaeda in of translocation to the extalimital regions. diapausing P. variegata.

PS03.38 Thelazia Callipaeda Eyeworm and PS03.39 Detection of Theileria Orientalis Its Vector Phortica Variegata: Strategies to Genotypes and Identification of Potential Survive the Winter Vectors in Central Queensland

Arme Thomas1,2, Riccardo P. Lia2, Marco Miss Kacie Baker1, Mr. Biniam Lakew1, Dr. Pombi3, Domenico Otranto2 Sarbast Kheravii1, Mr. Grant Parker2, Professor 1School of Biological Sciences, University Steve Walkden-Brown1 of Leicester, Leicester, United Kingdom, 1University Of New England, Armidale, 2Dipartimento di Medicina Veterinaria, Australia, 2Department of Agriculture and Università degli Studi di Bari, Valenzano Fisheries, Emerald, Australia Bari, Italy, 3Dipartimento di Sanita Pubblica e Malattie Infettive, Sapienza Università di Theileria orientalis is a tick-borne protozoan Roma, Rome, Italy parasite of bovine red blood cells inducing persistent infection. Clinical theileriosis, Thelazia callipaeda (Spirurida, Thelaziidae) caused by T. orientalis has become a cattle is a vector-borne nematode that infects disease of economic importance in Australia the conjunctival sacs and associated orbital since 2006 with the emergence of pathogenic tissues of many mammals, including humans. genotypes of the parasite. There is a scarcity The nematode was once commonly referred of information on the identification of these to as the ‘Oriental eyeworm’, but after more genotypes and their epidemiology in central than 20 years, it has spread throughout Queensland. We used recently developed Europe and many studies have elucidated molecular methods to differentiate and the role of a bizarre fruit fly (i.e., Phortica quantify pathogenic and non-pathogenic T. variegata) as the intermediate host and orientalis genotypes in cattle and ticks on a vector of this zoonotic nematode, through property near Capella, Central Queensland its lacriphagous behaviour. Though domestic where clinical theileriosis is believed to be and wild carnivores are the main definitive absent. Thirty blood samples were collected hosts for this parasite, more than 1000 and 10 from cattle and ticks were collected human cases have been reported, respectively, off pasture and off animals fortnightly in Asia and in Europe. The presence of adult between February and June 2018. Real-time nematodes and their serrated cuticle cause quantitative PCR (qPCR) analysis of the a variety of clinical signs in the infected blood samples revealed a generic T. orientalis animals which range from mild conjunctivitis, prevalence of 100% with prevalence of the follicular hypertrophy of the conjunctiva, different genotypes being 100% for Buffeli foreign body sensation, epiphora, itchiness, (non-pathogenic), 80% for Chitose (possibly congestion, swelling and even blindness. The pathogenic) and 0% for the pathogenic Ikeda close relationship among T. callipaeda, their genotype. Absolute quantification revealed definitive hosts and their vector P. variegata similar parasite burdens of Buffeli and Chitose remains in many instances enigmatic and, with no association between the two. It also unlike many other arthropod vectors, flies revealed significantly higher Theileria burdens feeding around the eyes of definitive hosts in sampled cows than heifers. On pasture, (therefore transmitting the third infective two tick types were detected, Amblyomma larval stages) are almost exclusively males. sp. (95.8%) and Haemaphysalis bancrofti Indeed, the development of the nematode (4.2%). The ticks collected from cattle were larval stages (L1-3) closely associated with the very different being 98.6% Rhipicephalus biology of the male. In this poster the authors microplus and 1.4% Amblyomma triguttatum. will describe recent field and laboratory Quantitative PCR of pooled tick head parts of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 300 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

the three tick species revealed high levels of these ones the cause of diarrhea in all clinical genotype Buffeli in H. bancrofti and moderate cases. A significant association (p = 0.007) levels in R. microplus. No other Theileria among calves with diarrhea and high OPG genotypes were detected. Based on these was found. There was a positive association and other findings the most likely vector of between number of oocysts (≥4000 OPG) T. orientalis in this area is Haemaphysalis and diarrhea. On the other hand, the risk of bancrofti despite the lack of detection on diarrhea increases (Relative Risk = 2.37) when cattle. Further research is required on the the number of oocysts eliminated increases role of Rhipicephalus microplus. T. orientalis too. The presence of E. bovis and E. zuerni was endemic on this property but the most in cases of diarrhea shows that this disease pathogenic genotype, Ikeda was absent. is being underestimated in calf rearing. It is recommended to maximize hygienic-sanitary measures in these dairy farms. The high PS03.40 Eimeria Spp. And Its Association incidence of calves’ infections suggests that With Diarrhea in Dairy Calves in Uruguay fecal monitoring is a necessary tool for the control and prevention of this parasite disease. Licenciado en Laboratorio Anderson Saravia1,2, Doctor of Veterinary Science Franklin Riet Correa1, Doctor of Veterinary PS03.41 Mitochondrial DNA of Babesia Science Eleonor Castro-Janer2 Odocoilei as a Means of Diagnosis for 1Instituto Nacional de Investigación Babesiosis in Cervids Agropecuaria (INIA). Estación Experimental INIA La Estanzuela, Colonia del Sacramento, Ms. Fiorella Vialard1, Mr. Alexandre Léveillé1, Uruguay, 2Facultad de Veterinaria, Universidad Dr. Adriana Pastor2, Dr. John Barta1 de la República, Montevideo, Uruguay 1University of Guelph, Guelph, Canada, 2Calgary Zoo, Calgary, Canada Bovine eimeriosis is a disease of worldwide distribution that cause significant economic Babesia odocoilei is a piroplasm losses. It is produced by Eimeria spp. and (Apicomplexa) infecting cervids that is affects young animals, mainly. Many species transmitted through blood-feeding by its have been described but only few are tick definitive host, Ixodes scapularis. In pathogenic. Usually, it is a self-limiting disease. dead-end cervid hosts (e.g. reindeer and elk), B. odocoilei can cause hemolytic In Uruguay, only two cases of eimeriosis have anemia and death. Although frequently been reported, one by Eimeria zuernii (1918) used for species identifications, nuclear and, recently, other by Eimeria bovis (2015). 18S ribosomal DNA (rDNA) sequences may Although several suspects were reported, lack sufficient species-level divergence to little is known about the different species provide reliable identifications. Consequently, that could affect cattle and which are more diagnostic tests based on this locus may lack relevant in clinical cases. The objectives of this the ability to differentiate among closely study were to identify the Eimeria species in related piroplasms. In contrast, piroplasm feces of 757 calves (<10-month-old) from ten mitochondrial (mt) DNA exhibits extensive different dairy farms with diarrhea in calves species-level diversity that can be exploited and to determine their responsibility as a for diagnostics. Complete mt genome cause of diarrhea. sequences of related piroplasms (e.g. Babesia gibsoni, Babesia bigemina, Babesia bovis, and The oocyst count per gram (OPG) was Babesia caballi) are available but not for B. determined by McMaster method and for odocoilei. We used polymerase chain reaction identification species, fecal sample cultures (PCR) and amplicon sequencing to generate to sporulation were used. Six species were a substantial majority (>5.3 kbp) of the mt identified: E. bovis, E. zuernii, Eimeria genome of B. odocoilei. DNA was obtained ellipsoidalis, Eimeria auburnensis, Eimeria from the spleen of a reindeer confirmed canadensis and Eimeria alabamensis, with a to have died from a B. odocoilei infection. high prevalence of E. bovis and E. zuernii being Standard PCR amplifications were performed

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 301 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

using primers targeting conserved regions of PS03.43 Repellent Effects of Encapsulated apicomplexan mt genomes. PCR amplicons Carvacrol on the Southern Cattle Tick, were purified and Sanger sequenced. The Rhipicephalus (Boophilus) Microplus (Acari: mt genome has the same components and Ixodidae) arrangement as other related piroplasms including three protein-coding regions (CDS; Aldilene Silva Lima1, Gabriel Alves Landulfo1, i.e. cytochrome c oxidase I [COI], cytochrome Livio Costa-junior1 c oxidase III [COIII], and cytochrome B 1Federal University of Maranhão, São Luis, Brazil [CytB]) and numerous fragmented rDNA. Unlike the nuclear 18S rDNA sequences, the Rhipicephalus (Boophilus) microplus mt CDS regions demonstrated considerable (Canestrini) is a problem for livestock genetic divergence among these related production systems, and its control has piroplasms. For example, pairwise sequence become challenging due to the selection identities of partial COI sequences ranged of tick populations resistant to synthetic from 76 to 82% among the related Babesia chemical acaricides. The use of repellent species listed above. This newly generated B. compounds prevents contact between the odocoilei mt genome sequence will provide a arthropod parasite and the host and can thus target for more reliable species delimitation, contribute to increases in the efficacy of these potentially including species specific PCR, acaricides. Carvacrol monoterpenic phenol is than the existing nuclear 18S rDNA sequences a possible alternative method for controlling and will support the detection of babesiosis in R. (B.) microplus; however, this compound wild and farmed cervids. is highly volatile, and its volatilization can be decreased through microencapsulation, which results in the timed release of the PS03.42 Metabolic Budget of Ticks in The compound. The cell wall of Saccharomyces Field in S.W. England cerevisiae can be utilized for the protection of volatile molecules. The aim of this study Mr. Saeed Alasmari1, Professor Richard Wall1 was to evaluate the in vitro repellent effect 1University of Bristol, Bristol, United Kingdom of yeast cell wall-encapsulated carvacrol on susceptible R. (B.) microplus larvae. The spread of tick-borne diseases in many Specifically, the vertical filter paper bioassay parts of the world represents a significant was employed to analyze the repellent activity health challenge for livestock and companion of encapsulated carvacrol, nonencapsulated animals despite many advancements in the carvacrol and DEET(N,N-diethyl-meta- understanding of tick biology. Of crucial toluamide) at concentrations ranging from importance to the success of a tick species 0.75 to 0.001mg/cm², and the repellent to reproduce and thrive in its specific activities were evaluated. Both carvacrol environmental niche is the ability to efficiently and encapsulated carvacrol exhibited obtain, store and metabolise the key bio- repellent effects on R.(B.)microplus larvae, nutrients from their blood meal. Neither the and the encapsulated compound showed metabolic requirements of developing ticks the highest repellent activities at the lowest nor the allocation of resources to development concentrations. Carvacrol encapsulated have been studied extensively. Therefore, many exhibited a low repellent concentration in all aspects of tick physiology remain unclear. times (≤0.05mg/cm2), while the carvacrol In the present work, a range of biochemical nonencapsulated ranged CR50 from 0.13 to assays were used to estimate the seasonal 0.27mg/cm2 at 1 to 6h posttreatment. The patterns of free sugar (glucose), glycogen, present paper provides the first description lipid and protein accumulation through the life of the use of a microencapsulation technique cycle of ticks in s.w. England. A total of 1.303 for achieving the highest repellent effect of nymphs, males and females were analysed over carvacrol and indicates that this technique the course of a year. The metabolic patterns might be used to obtain new delivery systems observed provide a direct insight into the for volatile and hydrophobic compounds. feeding history and life-history partitioning of resource during tick development.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 302 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.44 Distribution of Ticks and Molecular PS03.45 The Use of Computed X-ray Detection of Tick-Borne Diseases From Dogs Microtomography (microCT) for Non- in the Republic of Korea, 2017–2018 invasive Imaging of Ixodes Ricinus Ticks

Jinhyeong Noh1, Hyunkyoung Lee2, Mi-Sun Adam Novobilsky1, Jaroslav Ondrus2 Yoo 1, Yeojin Park1, Subin Min1, Jiyeon Lim1, 1Veterinary Research Institute, Brno, Czech Bang-Hun Hyun1, Yun Sang Cho1 Republic, 2Department of Biology and 1Parasitic and Insect disease laboratory, Animal Wildlife Diseases, University of Veterinary and and Plant Quarantine Agency, Gimcheon-si, Pharmaceutical Sciences and CEITEC, VFU, Republic of Korea, 2Animal Pathodiagnostic Brno, Czech Republic laboratory, Animal and Plant Quarantine Agency, Gimcheon-si, Republic of Korea Candidatus Neoehrlichia mikurensis is an emerging bacteria transmitted by Ixodes Ticks are vectors of a number of pathogens ricinus in Europe. C. N. mikurensis can infect that are important to humans and also several mammals, including humans. However, veterinary practice. Many of the pathogenic the impact of C. N. mikurensis on internal agents transmitted by ticks, including morphology of I. ricinus is not known. Ehrlichia spp., Anaplasma spp., Borrelia Internal and external structure of ticks is spp., and Babesia spp., are known to be commonly visualized using light, electron humans and animal pathogens worldwide. microscopy or histology techniques. Recently, In this study, we are proposed to detect and computed X-ray tomography (microCT) has characterize tick-borne diseases in ticks from been used as a novel tool for investigation of Korea using molecular techiniques. morphologic features in insect. In comparison to electron microscopy and histology, Ticks were removed from shelter and microCT enables fast and easy sample companion dogs, also colllected near the processing, examination of several specimen animal shelterby dragging and flagging. Based at the same time. While final resolution of on microscopic examination, collected ticks images from microCT scans is sufficient (2.5 were identified to species and developmental to 6 m), major challenge of X-ray scanning stage characterized. DNA purified from ticks consists in low contrast of internal structures. was used for the detection of 6 of tick-borne Aim μof the study was to develop and diseases (Anaplasma phagocytophilum, A. optimize protocol for microCT scanning platys, Ehrlichia chaffeensis, E. canis, Borrelia of both unfed and fully fed ticks of I. spp., Babesia spp.) using previously described ricinus. Unfed ticks were dehydrated in a PCR assays. graded 70-100% ethanol, then incubated in 2% osmium tetraoxide and dried using A total of 2,294 ticks from two genera hexamethyldisilazane. Another group of and three species (2,261 of Haemaphysalis unfed ticks were dried by letting them placed longicornis, 10 of H. flava, 22 of Ixodes at room temperature for several weeks. All nipponensis) was collected from northern ticks were scanned on Bruker Skyscan 1276 area (1,461 ticks), central area (718 ticks) and with no filter, resolution 4 m and 4K binning. southern area (115 ticks) of ROK. 24 of 1,111 Scanned data were reconstructed using tick pools (2.2%) were PCR-positive for A. N-recon, further modifiedμ and analyzed using phagocytophilum, and 4 of 1,111 tick pools CTan software. 3-D rendered volumes of ticks (0.9%) PCR-positive for Borrelia spp. were visualized in CTvox. Contrast and quality of internal structure did not differ between In this study, tick identification and molecular naturally dried and ethanol dehydrated ticks detection were conducted to monitor of ticks contrasted with osmium. In group of fed ticks, and tick-borne diseases in the Republic of artificial in vitro blood feeding of the ticks Korea. It is important to continue the efforts with addition metal-based contrast agents will to identify additional tick-borne pathogens in be carried out as next step. In conclusion, our ticks because of public health significance of study shows that microCT is a novel efficient these agents. tool for imaging of tick internal structure.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 303 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

This study was supported by OP VVV microscopic and molecular technique was Project CZ.02.1.01/0.0/0.0/15_003/0000 9.29% and 19.0% respectively. The infection 495 FIT (Pharmacology, Immunotherapy, was higher in males 6.05% than females nanoToxicology). 3.24%. Younger equids (<12 years) were more infected 6.91% than older ones (>12 years) 2.38%. The Arewa 4.7% and 6.42% Idabari PS03.46 Ticks and Piroplasms of Equines in breeds of horses and donkeys respectively Nigeria: Prevalence, Risk Factors and Health were the most infected. Implications.

Thankgod Onyiche1, Prof. Oriel Thekisoe1 PS03.47 Cross Border Transhumance, a 1North West University Potchefstrrom, South Dissemination Way of Ticks and Tick-Borne Africa, Potchefstroom, South Africa Pathogens in West Africa

The aim of the study was to conduct an Achille S. Ouedrogo1, Olivier Zannou5, epidemiological investigation of ticks and Souaibou Farougou3, Adrien Belem4, Claude equine piroplasms by determining the species Saeger5, Marinda C. Oosthuizen6, Abel diversity of ticks, detection of piroplasms, Biguezoton2, Laetitia Lempereur1 risk factors associated with prevalence and 1Laboratory of Parasitology and Parasitic the possible effects of these pathogens on Diseases, Center for Fundamental and Applied the health of equines in Nigeria. Ticks and Research for Animal and Health (FARAH), blood samples were collected and counted Faculty of Veterinary Medicine, University from 463 equids (horses and donkeys) in of Liège, Liège, Belgium, 2Unité Maladies à four (4) states, namely, Kano (n =169), Jigawa Vecteur et Biodiversité (UMaVeB), Centre (n=139), Katsina (n = 108) and Kaduna (n International de Recherche-Développement = 47). Epidemiological questionnaire was sur l’Élevage en zone Subhumide (CIRDES), also administered to the animal owners to Bobo-Dioulasso, Burkina Faso, 3Unité de ascertain risk factors. A total of 445 ticks Recherche sur les Maladies Transmissibles representing four species were collected. (URMAT), Université d’Abomey-Calavi, The overall prevalence of tick infested horses Abomey-Calavi, Bénin, 4Laboratoire de was 24.6% while that of donkeys was 0.5%. Santé Animale Tropicale, Institut du Rhipicephalus evertsi evertsi 85.84% was Développement Rural, Université Nazi BONI, the most abundant, followed by Hyalomma Bobo-Dioulasso, Burkina Faso, 5Research Unit truncatum 8.76%, Hyalomma dromedarii 3.59% in Epidemiology and Risk Analysis applied to while Rhipicephalus decolaratus 1.79% was the veterinary sciences (UREAR-ULg), Center for least abundant. The average tick burden was Fundamental and Applied Research for Animal 6.5 with a range of 1-14. The highest collection and Health (FARAH), Faculty of Veterinary of ticks was from Kano state 32.35% while Medicine, University of Liège, Liège, Belgium, Kaduna state15.06% was the least. 6Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Analysis of the risk factors associated with Pretoria, Pretoria, South Africa tick infestation reveals that the Arewa breed of horses has the greatest risk 15.9% of been The extensive and low-input livestock system infested with ticks. Other breeds include Talon occurring in West Africa lay on a cross-border 4.71% and Sudanese 3.99%. Male horses had transhumance which represents an important higher tick infestations 16.5% than females animal production strategy. Meanwhile, cattle 8.69%. Furthermore, younger horses (<12 tick infestations constitute one of the main years) 21.38% had higher infestations than constraints to the development of livestock older horses (>12 years) 3.62%. production in this region. Pathogens involved can be spread by live animals or vectors Microscopic and molecular analysis revealed movements. This study aims to evaluate the the presence of piroplasms such as Babesia involvement of transhumance in the spread caballi and Theileria equi. The overall of ticks and associated pathogens between prevalence of equine piroplasmosis using the eastern Burkina Faso (BF) and northern

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 304 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Benin (BN), where the sanitary conditions and an increasing incidence of these diseases of cattle, considering ticks and tick-borne is the most significant potential outcome diseases, are less known. The first step has of climate changes that affect the ticks been a cross sectional survey, which consisted directly or indirectly. Transmission of infection in an investigation on ticks and tick-borne occurs when there is an overlap of activities pathogens in cattle in the Eastern BF (490 between reservoir, vectors and humans, cattle) and the Northern BN (456 cattle). and differs according to the pathogens and The second step was a longitudinal survey location. Climate change may impact all including 311 cattle from BF, monitored of these stages and their interactions. The during one season of transhumance. These changes in climate and the length of different two surveys were carried out with cattle seasons directly affect tick survival, activity ticks and peripheral blood collection from and development because of the concept December 2016 to March 2017. According to that rising temperature will result in greater the cross sectional survey, the most abundant abundance of ticks. Additionally, climatic tick species recorded during this period change will also have indirect effect on tick were Amblyomma variegatum, Hyalomma borne pathogen transmission by affecting the truncatum and Rhipicephalus (Boophilus) survival and abundance of tick maintenance microplus. This latter species was evidenced hosts. The magnitude of the effects of climate in some areas in Northern BN where it wasn’t change in an endemic area depends on local reported yet, and only in a herd of BF coming conditions, ecosystems and biodiversity, from transhumance in BN. The microscopic migration patterns of birds and immunity in examination of blood smears revealed a high the population. There may be difficulties in prevalence of Anaplasma sp up to 86,12% determining the future scenario, however, it and 85,75% respectively in BF and BN. The can be overcome by conducting long-term genus Theileria was solely diagnosed in BN studies regarding disease incidence, tick within 8,11% of cattle. Fisher exact test showed biology, tick distribution and tick abundance, a prevalence of Babesia significantly higher host abundance and distribution and relevant (p<0.05) in BN (36.84%) than in Burkina Faso vegetation biology, specifically in relation to (5.31%). Such pattern of ticks and tick-borne climate change. pathogens distribution suggest a south- north expansion gradient which will be more investigated using longitudinal survey data PS03.49 Development and Evaluation of and by Reverse Line Blot hybridization essay. an Innovative Product Formulated With Essential Oils for Rhipicephalus Microplus Tick Control PS03.48 Threat of Ticks and Tick Borne Diseases in the Scenario of Changing Climate Student Leandro Rodrigues1, Dra. Cecília José Veríssimo1, Germano Scholze1, Dra. Luciana Rizwan HM1, Sajid MS2, Haider Abbas1, Mahvish Morita Katiki1, Dr. Rodrigo Giglioti1 Maqbool2 1Institute of Animal Science, Nova Odessa, 1Discipline of Parasitology, KBCMA College Brazil of Veterinary and Animal Sciences, Narowal Sub Campus UVAS Lahore, Narowal, Pakistan, Rhipicephalus microplus tick causes a lot of 2Department of Parasitology, University of damage to susceptible cattle. Its control is Agriculture, Faisalabad, Pakistan increasingly difficult due to the resistance to acaricides. Some plants naturally contain Zoonotic tick-borne diseases have become active principles to protect them from insect increasingly prominent and the incidence attack, among them are the essential oils. The of these diseases have risen dramatically Institute of Animal Science, in partnership over the last decade. Climatic suitability for with HYG Systems, has developed an essential a tick population is the fitness of a set of oils based product (HYGIZ) that has been climatic conditions for the existence of that tested in vitro and in vivo. In the in vitro tests, population in a given region. All the tick performed in the Laboratory of Parasitology species are important vectors of diseases of the Institute of Animal Science in Nova

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 305 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Odessa, SP, the product presented excellent as positive controls and, as negative controls, results: 100% efficacy in adult immersion test; water and alcohol/acetone (1:1). In order 100% repellency in 72 hours; 100% mortality to verify the acaricide odor effect on ticks, in larvae immersion test and odor test. Two it was created an open experimental unit in vivo tests were performed. In the first one, composed by two Falcon tubes connected animals treated with the HYGIZ had a drastic and interleaved with filter paper to prevent drop (more than 80%) in the tick count made the direct contact of the ticks with the disc in the first week of the experiment already impregnated with the oils that stayed in the and in the follow ones; unlike the group inferior tube. An opening was made in the which received a comercial Pour on product plastic cover of the upper tube that was shut that only droped tick counts in the following with filter paper to allow only the odor to counts (+14 and +21 days). In the other in vivo escape to the environment. In each treatment, test, teleogines were counted throughout the larvae or engorged females were exposed to body of 10 animals on days -2, -1, 0 in order the odor for 72 hours. In the bioassays with to distribute the animals homogeneously in engorged females, the oviposition inhibition, two groups: treated or not, and evaluated for percentage of hatched larvae, and product 22 consecutive days. After application, with efficacy were determined. Efficacies ranged a hand pump in the most infested regions 80-100% for almost all essential oils and its (neck, barb, arms, belly, groin, perineum, combinations in certain doses. Citronellal, scrotal sac and inside and outside the ears), 1,8-cineole and its combinations caused larvae HYGIZ controled all tick stages, and the daily mortality > 90%. In general, the odor of all effectiveness of the product ranged from 73.6 essential oils and their combinations had to 98.5%, with an overall effectiveness of 91%. acaricide effect on both larvae and engorged HYGIZ did not alter parameters related to females. The odor of commercial acaricides hepatic and renal functions of treated animals. had no effect on larvae and teleogines. (Sponsor: Hyg Systems).

PS03.50 Acaricidal Effect by Odors of Eucalyptus Globulus, Corymbia Citriodora, PS03.51 Estimates of Correlations and 1,8-Cineole and Citronellal Essential Oils Repeatability Between Infection Level of Against Rhipicephalus Microplus Tick Fever and Tick Infestation in Holstein Calves Raised in Paddock Leandro Rodrigues1, Mariana Santos Miranda1, Poliana A Pacheco1, Luciana M Katiki1, Rodrigo Rodrigo Giglioti1, Marco Antonio Faria Silva2, Giglioti1, Karen T Akashi1, Cecília J Veríssimo1 Luciandra Macedo Toledo1, Enilson Geraldo 1Institute Of Animal Science, Nova Odessa, Ribeiro1, Cecília José Veríssimo1, Cristiane Brazil, 2State University of Maringá, Maringá, F. Carvalho Fiorin1, Bianca Tainá Azevedo1, Brazil Guilherme Fávero Luciani1, Mariana Fogale Andrade1, Gunta Gutmanis1, Luciana Morita Essential oils have been investigated as Katiki1, Aníbal Vercesi Filho1, Cintia Hiromi an alternative for parasite control, aiming Okino3, Marcia Cristina de Sena Oliveira3 to delay the development of tick genetic 1Institute Of Animal Science, Nova Odessa, resistance, a consequence of the extensive Brazil, 2Federal University of Jequitinhonha use of chemicals for its control. The present and Mucuri´s Valley, Diamantina, Brazil, study aimed to evaluate the acaricidal effect 3Embrapa Pecuária Sudeste, São Carlos, Brazil of the odor of Corymbia citriodora and Eucalyptus globulus essential oils and its main The present study aimed to estimate the constituents, citronellal and 1,8-cineole, at the repeatability and correlation between doses of 10, 20, 40, 60, 80 and 100 mg.g-1 the infection levels of Babesia bovis, B. and in binary mixtures (1:1) on Rhipicephalus bigemina and Anaplasma marginale and microplus larvae and engorged females. natural infestations by R. microplus in calves Commercial acaricides (amitraz 12.5%, reared individual with shelter in paddock. cypermethrin 15% and cypermethrin 30% + Twenty newborns calves were monitored chlorpyrifos 30% + fenthion 15%) were used biweekly in the period between 08/21/2018

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 306 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

and 10/30/2018 (5 evaluations), wherein Babesia spp and Theileria spp in cattle. They R. microplus adults females were counted kill humans and animals, and have many and blood samples were collected for DNA economic losses of cattle and poultry farms. extraction. The infections levels for each Human tried to control the ticks and poultry hemoparasite specie were estimated by mites very long times but it was very difficult. qPCR. The qPCR assays set primers for B. In these trials, we tried to evaluate some bovis and B. bigemina were located in a commercial insecticides and select some region of cytochrome B (mt-cyt B) gene, good effective drugs. Six anti-insecticide whereas for A. marginale were located in drugs, Cyper-Killer (A), CyperKill Power (B), surface major protein 1b (msp1b) gene. The New-Goldenpex EC (C), New-Goldenpex qPCR assays were performed using HOT Powder (D), Green Ticks(E), and Natural FIREPol EvaGreen® qPCR Supermix (Solis herb extracts(F) were evaluated the efficacy Biodyne) and Rotor Gene Q thermocycler against bovine ticks and poultry red mites (Qiagen). The hemoparasites loads and tick in the experimental laboratory, field poultry counts were transformed into log10 (n + 1) farms and Korean cattle farms. for normal distribution approximation and were analyzed by the SAS Mixed procedure, In the anti-insecticides effect of insecticides in which the model included the fixed effects against bovine ticks in vitro culture, drugs of evaluation, sex, hemoparasite species A, B, C and D were good effective against and interactions. Animal was included in ticks in vitro culture. In the anti-insecticides the model as a random effect.The estimated effect of insecticides against poultry red mites correlation coefficients between tick counts in vitro culture, drugs A and C were good and hemoparasites loads were low or close effective against poultry mites in vitro culture. to zero, as well as the correlations between In the anti-insecticides effect of insecticides A. marginale and Babesia spp loads. against bovine ticks in Korean cattle farms, However, the correlation between the two drugs A, B and D were good effective against species of babesias levels was 0.49. The ticks in field trials. In the anti-insecticides repeatability estimates of B. bigemina, B. effect of insecticides against poultry red mites bovis and A. marginale were 0.74, 0.77 and in the poultry farms, drugs C and F were best 0.70, respectively. Based on these results, effective against poultry red mites. And drugs under conditions of the present study, we B, D and E were good effective against them. suggest that it is possible to identify animals presenting a most resistant phenotype against So that drugs A, B and D were recommended infection by hemoparasites. Furthermore, the to be used in the bovine ticks and C and F high correlation found between both babesias were recommended to be used in the poultry indicates that the factors that determine an red mites. increase or decrease in parasitemia for one species cause the same variation in the other species. PS03.53 A High Throughput Deep Amplicon Sequencing Method to Show the Emergence and Spread of Calicophoron Daubneyi in PS03.52 Studies on the Insecticides Against United Kingdom Cattle Herds Hard Ticks and Poultry Mites Umer Chaudhry1, Kashif Shahzad2, Stella Hee-jeong Youn1 Mazeri1, Neil D. Sargison1 1College Of Veterinary Medicine, Seoul 1University Of Edinburgh, United Kingdom, National University, Seoul, South Korea 2University of Skovde, Sweden

Ticks and poultry mites are very fecund The prevalence of C. daubneyi infection and resistant to environment, so that we in the United Kingdom has increased, but can easily see them in pasture of cattle despite the potential for rumen flukes to and poultry house. They are bloodsucking cause production loss in ruminant livestock, and cruel to animals, and transmit many understanding of their emergence and spread kinds of pathogens to host animals, such as is poor. Here we describe the development

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 307 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

of a method to explore the multiplicity of C. daubneyi infection and patterns of the F. hepatica DNA was detected in silage parasite’s emergence and spread, based on samples inoculated with as few as 6 Illumina MiSeq deep sequencing of meta metacercariae using ITS-2 PCR. Metacercariae barcoded amplicons of a fragment of the mt- from laboratory-maintained G. truncatula COX-1 locus. Our results show high levels of specimens were shed onto grass samples of genetic diversity per infection and between 20%, 30% and 40% dry matter (DM) content. populations of 10 to 47 of adult C. daubneyi, Ensiling vessels were either sealed with an each from a total of 32 finished prime cattle anaerobic fermentation lock or perforated consigned to slaughter from northern United plastic wrap and ensiled for 2, 6 and 10 weeks. Kingdom; with 18 unique mt-COX-1 haplotypes. Following ensiling, PCR and in vitro assays This has implications for the adaptability of comparing excystment rates of ensiled and un- environmental and intermediate host stages ensiled control metacercariae were conducted. of the parasite to changing climatic and animal management conditions, or of parasitic F. hepatica DNA persisted at 2, 6 and 10-week stages to exposure to anthelmintic drugs; timepoints within silage originating from grass potentially allowing for greater pathogenicity, of all DM contents, under both anaerobic and or the development of anthelmintic resistance, aerobic conditions. In terms of excystment respectively. Our results illustrate the impact rates; following 2 weeks anaerobic ensiling of high levels of animal movements in the no excystment occurred irrespective of initial United Kingdom, whereby multiple common grass DM. However, under aerobic (spoiled) mt-COX-1 haplotypes were identified in 26 ensiling conditions, ~28% excystment populations in the absence of geographical occurred after 6 weeks (30% DM content) clustering of clades. and unhatched juvenile parasites developed following 10 weeks (20% DM content).

PS03.54 Can Fasciola hepatica Metacercariae In conclusion, whilst detection of F. hepatica Survive Within Grass Silage And Retain DNA in silage is useful, excysting metacercariae Viability? post-ensiling is a more useful indicator of viability. Anaerobic conditions are vital to Miss Bethan John1, Dr. David R Davies2, prevent survival of metacercariae, highlighting Professor Diana Williams1, Professor Jane the importance of forage sealing. These Hodgkinson1 investigations will lead to informed husbandry 1Institute of Infection and Global Health, The advice on reducing fasciolosis prevalence University of Liverpool, United Kingdom, through improved silage management. 2Silage Solutions Ltd, Ceredigion, United Kingdom PS03.55 Efficacy of Oral Afoxolaner Plus Fasciola hepatica, the common liver fluke, Milbemycin (NexGard Spectra®) on the is an important cause of morbidity and Reduction of Eggs per Gram (EPG) of Feces mortality in ruminant livestock worldwide. It is of Dogs Naturally Infected With Ancylostoma recognised that metacercariae can overwinter Sp., Toxocara Sp. And Trichuris Vulpis in Brazil on pasture and infect grazing livestock the following spring. However, the infection risk Andre Cutolo1, Luis Pelissoni1, Debora posed by feeding silage is not well described. Sartor1, Gabriela Vito1, Cristiano Nascimento2, Existing studies date back to 1961 and pre- Tassia Vale3, Isabella Souza3, Carolina Silva3, date the modern advantages of applying Livio Costa-Junior, Filipe Freitas4, Brenda molecular techniques to detect F. hepatica Deeckens4, Maycon Heidmann4, Bruno de DNA persistence within silage. The impact Castro4 of aerobic spoilage on survival of ensiled 1Boehringer Ingelheim Animal Health Do metacercariae and the ability of juvenile Brasil Ltda., Paulínia, Brazil, 2Convolution, parasites to excyst must be understood, to Ribeirão Preto, Brazil, 3Universidade Federal establish how silage provision impacts F. do Maranhão, São Luis, Brazil, 4Universidade hepatica transmission. Federal do Mato Grosso, Sinop, Brazil

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 308 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.56 In Vitro Hatchability of Eggs of Ancylostoma spp., Toxocara spp. and Trichuris Haemonchus Contortus and Trichostrongylus vulpis are the most prevalent nematodes Colubriformis With Powdered Bark of Acacia infecting dogs in Brazil. Infection by these Mearnsii worms may be asymptomatic but also lead to different lesions and blood/nutrient Murilo Antonio Fernandes1, Poliana A depletion in the gut depending on the level of Pacheco1, Jessica Carolina Ribeiro2, Daniele infestation. Ancylostoma spp. and Toxocara Rizatto Balbino2, Bruna Costa Rodrigues spp. are also important as zoonotic agents Wedy2, Dr. Sui Mui Tsai1, Dr. Luciana M. Katiki2, since L3 larvae may infect humans as larva Dr. Helder Louvandini1 migrans. NexGard Spectra® is a combination of 1University of São Paulo, Centre for Nuclear Afoxolaner and Milbemycin Oxime promoting Energy in Agriculture., Piracicaba, Brazil, a broad efficacy action versus canine ecto 2Institute of Animal Science, IZ/APTA, SAA - and endoparasites. Different breed owned SP, Nova Odessa, Brazil dogs, weighing greater than 2kg and more than 8 weeks old, from urban areas of São Gastrointestinal helminths in small ruminants Luis, Maranhão state, Sinop and Ipiranga do are one of the major health problems Norte, Mato Grosso state, were included in the affecting the production of these animals study if harboring quantifiable eggs of any worldwide. The resistance of these parasites of these worm species on a McMaster EPG to commercial anthelmintics entails great evaluation. Eighty-four Ancylostoma spp., 31 economic losses. Therefore, the search for Toxocara spp. and 21 Trichuris vulpis. infected effective alternatives for the control and dogs were included in the study and treated eradication of these helminths has been the once orally with a single tablet delivering target of researches, among which the use 2.50 to 5.2 mg/kg of afoxolaner + 0.50 to 1.07 of phytotherapeutic compounds. Among mg/kg of milbemycin oxyme per dog. Few these compounds, plants with high contents included animals harboured mixed infections. of condensed tannins, such as Acacia The individual EPG were transformed to Log mearnsii, have been presented as a promising (x + 1) and the efficacy was calculated based alternative in the control of these parasites. on the comparison of the geometric mean The objective of this study was to evaluate the number of EPG before treatment versus the effect of Acacia mearnsii powdered bark (AM) mean number at Day 14 (±4). The geometric on egg hatch assay (EHA) to Haemonchus means were calculated by averaging the EPG contortus and Trichostrongylus colubriformis. log-counts, taking the anti-logarithm, and Decreasing concentrations of the AM (50; then subtracting 1. The mean number of EPG 25; 12.5; 6.25; 3.125; 1.56; 0.78; 0.39; 0.19; pre-treatment and percentage of efficacy 0.09; 0.04; 0.02; 0.01; 0.006; 0.003; 0.001 were 1031 and 98.5% for Ancylostoma spp., mg/mL) was solubilized with Tween80 and 665 and 96.8% for Toxocara spp., and 400 distilled water, were added in 48 well plates, and 96.8% for Trichuris vulpis.. NexGard each well containing approximately 100 eggs Spectra® showed excellent efficacy results in of H. contortus and the same conditions for the field against Ancylostoma spp., Toxocara T. colubriformis and incubated for 24 hours spp. and Trichuris vulpis. in housed naturally at 27°C. The data were analyzed with the infected dogs in Brazil. software SAS® using the PROBIT procedure to estimate the LD50 (lethal dose enough to inhibit hatchability of 50%. The LD50 for T. colubriformis eggs was 1.28 mg/mL, and for H. contortus eggs was 13.55 mg/mL. Thus, we can conclude that AM may be a potential alternative for the control of gastrointestinal parasites in small ruminants, due to inhibitory effect showed on egg hatchability, being more effective in T. colubriformis eggs, once it presented a lower LD50.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 309 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.57 Effect of Different Faecal mm/d). Albendazole reduced the mycelial Concentrations of Anthelmintics on Mycelial growth only at 1 ppm (0.89 mm/d, P<0.0001), Growth of the Biological Control Agent while fenbendazole affected negatively the Duddingtonia Flagrans fungal growth in all concentrations (0.46 to 2.19 mm/d, P<0.0001). These results Sara Zegbi1, Dr Carlos Saumell1, Dr. Federica represent a first step on elucidating whether Sagües1, Dr. Laura Ceballos2, Inés Guerrero1, anthelmintics could be used in very specific Prof. Lucía Iglesias1, Dr. Silvina Fernández1 situations while biological control is applied. 1Área de Parasitología y Enfermedades Parasitarias, Centro de Investigación Veterinaria de Tandil (CIVETAN) (UNCPBA- PS03.58 Evaluation of Attractiveness and CICPBA-CONICET), Facultad de Ciencias Specificity of New Plastic Blue Screens Veterinarias, UNCPBA, Tandil, Argentina, for Monitoring and Control of Stomoxys 2Laboratorio de Farmacología, Centro de Calcitrans Populations Investigación Veterinaria de Tandil (CIVETAN) (UNCPBA-CICPBA-CONICET), Facultad Shukri Sharif3, Gerard DuvalletT2, Lucas de Ciencias Veterinarias, UNCPBA, Tandil, Etienne2, Clement Mongellaz4, Emmanuel Argentina Liénard1, Christelle Grisez1, Michel Franc1, Emilie Bouhsira1, Marc Desquesnes5, Philippe Duddingtonia flagrans acts as biocontrol Jacquiet1 agent by preying on pre-parasitic nematode 1National Veterinary School of Toulouse, larvae in animal faeces. This fungus could be Envt, Toulouse, France, 2UMR5175, University exposed to anthelmintic drugs eliminated in Paul-Valéry Montpellier, Montpellier, France, faeces, but little is known as to whether these 3UMR 1225- IHAP, INRA/ENVT, Toulouse, drugs could alter the fungal development France, 4VetAgro Sup, Lyon, France, 5CIRAD and its efficacy. Thus, the aim of this study Kasetsart, Bangkok, Thaïland was to determine the in vitro effect of certain anthelmintics on the growth of this fungus. Stomoxys calcitrans is considered as a major In two assays, active ingredients diluted in pest of livestock worldwide. Insecticides have methanol of the five anthelmintic drugs most been extensively used to control this pest but commonly used in Argentina were used in resistance to these chemical compounds is concentrations reported as found in bovine now reported in many countries. Therefore, faeces: levamisole, 1 ppm; albendazole, 0.027, a more sustainable and efficient control is 0.054 and 1 ppm; fenbendazole, 0.027, 0.054 needed. A lot of different traps have been and 1 ppm; ricobendazole, 1 and 2.77 ppm; tested to catch stable flies such as Vavoua and ivermectin, 1, 2 and 10 ppm. Each of these Nzi traps or Alsynite sticky traps. However, drug concentrations were added to corn meal low numbers of stable flies are caught per day agar (CMA) 2% and then poured on Petri and per trap with these devices (usually less dishes (n=12/concentration). Plates with CMA than two hundreds individuals per day and and containing only methanol and only D. per trap), which is sufficient for establishing flagrans were used as control. All plates were a population’s dynamics but not for control. inoculated with 1 cm2 of fresh D. flagrans The objective of this study was to evaluate mycelia growing in CMA, and incubated at the attractiveness and the specificity of seven 27ºC for 7 (assay 1) or 12 (assay 2) days. The different new types of blue screens (one fungal growth rate was determined every polyester fabric blue screen and six plastic 24 h by measuring the radial growth. Similar blue screens) for stable flies. These screens, fungal growth was obtained from the control showing slight differences in their reflectance plates containing only D. flagrans (6.79 to around 460 nm, were tested during summer 8.12 mm/d), only methanol (6.5 to 7.17 mm/d), 2016 in southwestern France. Height of the levamisole (6.49 to 6.99 mm/d) and both screen and its east or west orientation were concentrations of ricobendazole (6.51 and also considered. High levels of S. calcitrans 7.53 mm/d). Ivermectin slowed significantly captures were recorded during this study (P=0.0181 – P=0.0351) the fungal growth in (from 141 to 7301 individuals per blue screen all tested concentrations (6.14, 5.67 and 6.29 and per day) whereas the numbers of

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 310 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

Tabanids and pollinator insects remained are known to be lethal to ticks. Simple Green® very low (less than 10 individual per screen (SG), a household cleaning product, has and per day). No significant difference in anecdotal information of insecticidal activity. attractiveness has been shown between the The present study aimed to determine different types of blue screens. The lower half efficacy of EIC3, EPN and SG against adult of the blue screens caught significantly more ticks on deer hides. In vitro bioassays with stable flies (70%) than the higher half (30%). adult and larval stages of SCFT were carried The “east” side of the screen was a little more out to establish doses of EIC3 and SG to be attractive (60% of stable flies) than the west used in the hides treatment. For the deer hide side. These results are highlighting the interest test, each hide (10 replicates) was inoculated of these blue screens not only to monitor with ten engorged female ticks and treated populations but also to control stable flies in with one liter of EIC3 (6.25%), EPN (50,000 cattle farms as some insecticides could be infective juveniles/ml), SG (100%) or water incorporated in the plastic material. (control). After treatment the hides were rolled and placed into plastic bags and held at 8oC for 12 hours. After 12 hours, ticks were PS03.59 Alternative Treatments for Southern incubated to allow for egg laying and larval Cattle Fever Tick, Rhipicephalus Microplus hatch determination. SG treatment resulted in (Acari: Ixodidae) Infested White-Tailed Deer low mortality rates both in vitro (10%) and in Hides the hides (27%). EN resulted in even lower e mortality (<15%). EIC3 was the most effective Guilherme Klafke1, Robert J. Miller1, John treatment tested against SCFT resulting in Goolsby1, Donald B. Thomas1, Denise Bonilla2, 100% mortality of larvae, 94% reduction of Hallie Hasel3, Roberta Duhaime3, Chelsea fertility of adults and 98% mortality of fully Pike4, T.R. Lansford4, Andy Schwartz4, engorged females in treated hides. EIC3 Adalberto Pérez de León5 can be used as an alternative to treat SCFT 1USDA ARS Cattle Fever Tick Research infested deer hides. Laboratory, Edinburg, United States, 2USDA APHIS VS Natural Resources Research Center, Fort Collins, United States, 3USDA APHIS PS03.60 Dog’s Parasite Control Practices by VS Cattle Fever Tick Eradication Program, Veterinarians in Brazil Laredo, United States, 4Texas Animal Health Commission, Austin, United States, 5USDA Tássia Lopes Vale1, Brazil Isabella Chaves ARS Knipling-Bushland U.S. Livestock Insects Sousa2, Caio Pavão Tavares1, Nobate Gomes3, Research Laboratory and Veterinary Pest Livio Martins Costa Junior4 Genomics Center, Kerrville, United States 1Graduate Program in Health Sciences, State University of Maranhão - UFMA, São Luís, Southern cattle fever tick (SCFT), Brasil, 2Graduate Program in Biotechnology, Rhipicephalus microplus, is the most State University of Maranhão - UFMA, São economically important invasive species of Luís, Brazil, 3Biological Sciences Course, ticks in the United States. SCFT parasitizes State University of Maranhão - UFMA, São cattle and also wild ungulates as the white- Luís, Brazil, 4Laboratory of Parasite Control, tailed deer (WTD), Odocoileus virginianus, Department of Pathology, State University of which can spread tick populations posing Maranhão – UFMA, São Luís, Brazil a threat to the cattle fiver tick eradication program (CFTEP). Hunter killed WTD and The aim of the present study was assess other game hides are inspected for ticks and dog’s parasite control practices by Vets treated with coumaphos before they leave from different regions of Brazil. The on- tick quarantined areas of Texas. Concerns line survey were performed with 403 Vets. over human intoxication have led the CFTEP From all professional surveyed 41% (n=168) to consider alternative treatment methods. routinely ask stool test being 21% every Essentria® IC3 (EIC3), a plant essential oils- six month, 16% yearly and 98% performed based product and entomopathogenic prophylactic treatment to helminth. The nematodes (EPN) (Steinernema carpocapse) most commonly antihelminth (AH) used

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 311 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

was the association of Benzimidalozoles+P Vaccinated animals responded favorably yrimidines+Pirazinoisoquinolone (33%) and to immunization; over the sampling period, Benzimidalozoles+Macrocyclic lactones+Pyri vaccinates shed 66% fewer eggs (when midines+Pirazinoisoquinolone (22%). The AH measured as mean cumulative faecal egg group are change by 57% of the Vets and 26% counts) than challenge control lambs. At already suspected the inefficiency, being the postmortem, vaccine recipients had a 70% Benzimidazoles the group most cited (23%). mean reduction in mean total worm burden The data showed that 47% of the vets change compared to the control group. These results routinely the drugs against ectoparasites and indicate that, in young lambs of this breed, 92% associate with environmental control. A the subunit vaccine is capable of inducing high percentage of surveyed (58%) reported vaccine -protective responses at an early a dog hypersensibility to collar for fleas and age. Acknowledgements: European Union’s ticks control. The Isoxazoline was the most Horizon 2020 Research and Innovation indicated group (40%). The Vets report programme under Grant Agreement No. inefficiency of different chemicals groups 635408 (PARAGONE). to tick control. The most report was Pirazol (63%), followed by Pyrethroids (8.2%) and Isoxazoline (5%). These results show the PS03.62 Anthelmintic Effect of Banana indiscriminate use of antiparasitic compounds Plantation Residues in Sheep and the reports of resistance accelerates the need for guidelines for treatment and control Paulo Henrique Selbmann Sampaio1, Daniel of dogs parasites in the veterinary clinic. Daza2, Luciana Morita Katiki3, Danilo Eduardo Rozane4, José Roberto Cavichiolo5, Kato Edna Tomiko Myiake2, Maria Ines Genovese PS03.61 A Teladorsagia Circumcincta Rodriguez2, Eduardo Harry Birgel Jr.6, Rodrigo Recombinant Vaccine Prototype Is Able to Martins Soares7, Lilian Gregory7, Fernando Protect Lambs of Canaria Hair Breed Sheep José Benesi7 1Animal Health Research Center, Biological Tara Pérez-Hernández1, Julia N. Hernández1, Institute, São Paulo, Brazil, 2Faculty of Cynthia Machín1, Yolanda Corripio-Miyar2, Pharmaceutical Sciences, University of Harry W. Wright2, Dan R.G. Price2, Tom N. São Paulo, São Paulo, Brazil, 3Institute of McNeilly2, Alasdair J. Nisbet2, Jacqueline B. Animal Science, Nova Odessa, Brazil, 4São Matthews2, Jorge F. González1 Paulo State University (Unesp), Campus of 1Universidad De Las Palmas De Gran Canaria, Registro, Registro, Brazil, 5Food Technology Arucas, Spain, 2Moredun Research Institute, Institute, Campinas, Brazil, 6Faculty of Animal United Kingdom Science and Food Engineering, University of São Paulo, Pirassununga, Brazil, 7Faculty Gastrointestinal nematodes (GIN) are a major of Veterinary Medicine and Animal Science, constraint on production in the small ruminant University of São Paulo, São Paulo, Brazil sector worldwide. As anthelmintic resistance is now widespread, alternative non-chemical Acetone extracts of leaves, pseudostems control strategies have become a priority. and bunch peduncles of the “Nanica” Recently, a recombinant vaccine prototype banana cultivar, a triploid clone (AAA) of the against Teladorsagia circumcincta has been Cavendish subgroup, significantly inhibited shown to be protective in Texel-cross sheep. Haemonchus contortus eggs from hatching The level of immunity stimulated by the in vitro, even when condensed tannins were vaccine varies between individuals and with suppressed with polyvinylpolypyrrolidone (p age. Previous studies suggest that Canaria < 0,0001). A further extraction was performed Hair Breed (CHB) sheep have an element of with n-butanol. Thin layer chromatography ‘natural resistance’ to GIN infection, including (TLC) was conducted with chloroform: in young lambs. Here, we tested the effect methanol: water (70: 30: 4) as solvent system of the T. circumcincta recombinant vaccine and vanillin – phosphoric acid as reagent. A in three-month-old CHB lambs subjected retention factor corresponding to syringin to a trickle challenge with this parasite. was obtained. Syringin is a phenylpropanoid,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 312 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

a group of phenolic compounds related to banana plant resistance against Introduction: The protozoa of the genus phytonematodes. Total phenolic contents of Giardia remain a common cause of diarrhea leaves, pseudostems and bunch peduncles in dogs and constitute a public health were 66.2±0.4; 30.9±0.4 and 47.1±0.7 mg of concern. The aim of this study was to gallic acid equivalents per 100 g of sample, evaluate the diagnostic performance of respectively. No alkaloids were detected by the immunochromatographic strip test TLC. Samples were negative for saponins, Speed™ Giardia (Virbac) compared to the producing neither persistent foam nor parasitological method of flotation, the hemolysis. Wet basis moisture contents were enzyme immunoassay (ProSpecT™ Giardia 89.01%; 93.00% and 94.28%, respectively. Microplate) and the polymerase chain reaction Crude protein was 17.47%; 12.47% and (PCR) for canine giardiosis. 8.68%, respectively. In spite of lower protein contents, bunch peduncles are easily available Materials and methods: Initially, fecal samples at the packing houses of banana plantations, from dogs with the typical clinical signs of so they were chosen for in vivo trials. Twenty giardiosis were tested with the strip test four crossbred sheep artificially infected with Speed™ Giardia and two groups of 50 dogs Haemonchus were blocked into four groups emerged, namely group A and group B with according to their fecal egg counts (FEC). positive and negative samples, respectively. Each group was assigned to treatment with Thereafter, all samples were examined by zinc fresh chopped bunch peduncles fed at 0, 10, sulfate 33.2% flotation, the ProSpecT™ Giardia 20 and 30% dry matter during 14 consecutive Microplate assay and PCR. The combination days. Diet was complemented with Brachiaria of the last two methods was considered as a hay ad libitum and mineral salt. Sheep gold standard during statistical analysis. developed a normocytic anemia at the end of the trial. FAMACHA scores did not correlate Results: Giardia cysts were not detected with to FEC under trial conditions. Chopped microscopy in 16 out of the 50 samples (32%) peduncles have significantly reduced FEC on of group A, neither in samples of group B. day +13 (p=0.0286) compared to those fed Eight out of 50 samples of group B (16%) 0%. Although long term studies and further were tested positive both with the ProSpecT™ phytochemical investigation are required, the Giardia Microplate assay and PCR. Fecal current results offer evidence for use of these examination with the strip test Speed™ Giardia plantation residues as an anthelmintic. was significantly more sensitive (86.2%, CI95 of 75.1% to 92.8%) than the parasitological method (58.6%, CI95 of 45.8% to 70.3%) PS03.63 Giardia Spp.: Diagnostic Evaluation and gave results comparable to those of of the Immunochromatographic Strip Test, the ProSpecT™ Giardia microplate assay. the Parasitological Method of Flotation, the The specificity was excellent (100%, CI95 of Enzyme Immunoassay and the Polymerase 91.6% to 100%) for both Speed™ Giardia and Chain Reaction parasitological method.

Isaia Symeonidou1, Athanasios Gelasakis2, Conclusions: The immunochromatographic Androulla Miliotou3, Athanasios Angelou1, strip test Speed™ Giardia (Virbac) is a quick, Konstantinos Arsenopoulos1, Sofia Loukeri4, easy to perform and efficient method. Coralie Ségalen4, Christelle Navarro4, Professor However, in case of a negative result in dogs Elias Papadopoulos1, Vanessa Chala4 with compatible clinical signs it is advised to 1Laboratory of Parasitology and Parasitic confirm by an enzyme immunoassay and PCR. Diseases, School of Veterinary Medicine, Aristotle University, Thessaloniki, Greece, 2Agricultural University, Athens, Greece, 3Laboratory of Pharmacology and Parasitic Diseases, School of Pharmacy, Aristotle University, Thessaloniki, Greece, 4Virbac, Global Medical Department, Carros, France

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 313 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.64 Cost-Benefit Analysis of Four residues in meat, and pressure on helminth Helminth Control Strategies in Naturally populations for selection of resistance. Infected Sheep on a Low Technification Therefore, it can be inferred that application Property of the FAMACHA© method significantly reduced costs, in comparison to other Dr. Willian Maciel1, Msc. Isabela Barbosa1, Dr. techniques, making it possible to indicate it as Breno Cruz1, Mr. Dalmo Quilis1, Miss Ana Paula a strategy with the best cost-benefit ratio. Oliveira5, Dr. Gustavo Felippelli1, Mr. João Gabriel Flórid4, Dr. Lucas Gomes1, Dr. Welber Lopes2, Miss Laura Giroto1, Dr. Vando Soares3, PS03.65 A Case of Chorioptic Mange in a Dr. Alvimar Costa1, Dr. Gilson Oliveira1 Dairy Farm in the Municipality of Descalvado, 1Universidade Estadual Paulista - UNESP, SP, Brazil Jaboticabal, Brazil, 2Universidade Federal de Goiás - UFG, Goiânia, Brazil, 3Universidade Paulo Henrique Selbmann Sampaio1, Claudia Brasil - UB, Descalvado, Brazil, 4Universidade Del Fava1, Eduardo Pires Macedo2, Isabella Paulista - UNIP, Ribeirão Preto, Brazil, Barboza de Almeida3,4, Karina Araújo dos 5Faculdade de Educação São Luís de Anjos3,4, Shirley Batista da Silva Araújo3,4, Jaboticabal - FESLJ, Jaboticabal, Brazil Fernanda Calvo Duarte1, Leonardo Costa Fiorini1, Márcia Cristina Mendes1 This research evaluated cost-benefits of four 1Animal Health Research Center, Biological helminth control strategies in ovines. Forty- Institute, São Paulo, Brazil, 2Boehringer eight matriarchs, naturally parasitized by Ingelheim Animal Health, Campinas, Brazil, gastrointestinal helminths, were distributed in 3Master’s Program in Health, Food and four groups: GI - animals with egg per gram Environmental Safety in Agribusiness, of feces (EPG) counts ≥ 1000 by Mini-FLOTAC Biological Institute, São Paulo, Brazil, evaluations were treated; GII - similar to GI, 4Coordination of Superior Level Staff but using McMaster analyzes; GIII – all animals Improvement (CAPES) scholarship, Finance treated every 56 days; GIV – deworming of Code 001, Brasília, Brazil animals with FAMACHA© degrees 3, 4 or 5. Coprocultures were performed on fecal Fourteen lactating cows in a dairy herd samples, to identify helminths genres, and of 300 Holstein Frisian cattle presented parasitological necropsies were performed bilateral crusty focal lesions on the base of in five animals that died. For cost-benefit the tail. Skin scrapings were collected for calculations, sums of expenses with direct examination after clarification with treatments, laboratorial exams and materials 10% potassium chloride. Tissue samples for sample collection were calculated. from the affected areas and from the According to results of coprocultures and adjacent healthy skin were collected for necropsies, Haemonchus and Trichostrongylus histopathological examination with the were the most prevalent amongst all 10 aid of disposable dermatological punches. diagnosed genres, representing 94.49% of Skin samples were immediately placed in total parasitic burdens. Statistical differences cassettes for fixation with 10% buffered in EPG counts between groups were observed formaldehyde. At the laboratory they were in only five dates, with GIV showing higher dehydrated with increasing ethyl alcohol averages in three of those. Regarding concentrations and then treated with treatments, only GIII and GIV were similar (P xylol for later paraffin inclusion and block > 0.05), receiving a smaller number of doses assembly. Samples were cut in a microtome at the end of one year. Total treatment costs (3 micrometers) and assembled on slides were USD 2.74, USD 3.57, USD 4.65 and USD to be stained with hematoxylin and eosin. 3.82 for groups IV, III, I and II, respectively. Direct examination revealed Chorioptes Strategies I, II and IV required extra expenses bovis in all of the samples. Histopathological of USD 1,462.11, USD 1,391.71 and USD 11.89, examination revealed intense hyperkeratosis respectively. In spite of these extras, the and acanthosis of the epidermal strata; crusts strategy adopted in group III presents with acari and polymorphonuclear cells; disadvantages in relation to presence of drug intense multifocal eosinophilic, neutrophilic

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 314 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

and lymphoplasmacytic infiltrate in the propagation remains challenging. To expand epidermis and dermis; sudoriparous glands the diagnostic capabilities of our lab, we with dilated ducts; accentuated edema and have validated the novel application of an congestion of the epidermis and dermis; automated platform and assay developed moderate eosinophilic and lymphoplasmacytic for human diagnostics. Using 312 previously perivascular cuffing in the reticular dermis, MAT IgG antibody tested frozen serum consistent with an eosinophilic epidermidis samples (Negative=154; Positive=158), caused by parasitic mites. Infected animals we evaluated the miniVIDAS® and TOXO were treated with 0.5% eprinomectin pour Competition (TXC) assay (bioMerieux) for on (Eprinex Pour-on, Boehringer Ingelheim). comparison. Representative animal groups Except for one cow, all skin scrapings included our most common clinical and performed thirty days after treatment were research submissions: canine (n=62), feline negative. However, on this occasion, another (n=63), macropod (n=62), primate (n=61), two animals with clinical signs were identified. and aquatic mammal (n=64). Nonparametric These findings are an evidence of the usual analyses evaluated interrater agreement ( ), mild nature of this parasitic infection. Skin homogeneity ( 2), and performance (ROC lesions were restricted to the base of the AUC). Overall, TXC was an excellent test 횱 tail and didn’t result in apparent pruritus or compared to the훘 MAT for combined animal decreased productivity noticeable by the groups. Stratified analyses indicated that each farmer. The recurrence after a single treatment canine, feline, macropod, and primate animal with eprinomectin and the occurrence of group had near perfect or perfect agreement another two cases in the period exposes the with excellent performance. However, among difficulty of eradicating chorioptic mange, aquatic mammals, TXC had a poor to fair provided it seems to go unnoticed amidst the performance depending on the species. In this intense dairy farming routines. study, we determined that the novel use of the TXC assay has excellent overall performance for IgG detection, particularly in canines, PS03.66 A Novel Application of Enzyme- felines, macropods and primates. However, Linked Fluorescent Assay for Detection of additional work is required to validate this Veterinary Toxoplasma Gondii Infections in technique for aquatic mammals. This assay Multiple Hosts represents a valuable tool for detection of T. gondii in companion animals, captive exotics, Aly Chapman1, Heidi Wyrosdick1, John and wildlife. Schaefer1 1University Of Tennessee, Knoxville, United States PS03.67 Seroprevalence, Biogeographic Distribution and Risk Factors for Toxoplasma gondii is a global zoonotic Aelurostrongylus Abstrusus Infections in Cats protozoan parasite of endothermic animals. In addition to causing clinical disease in domestic Dr. Emily Katharina Gueldner1, Urs Gilli2, Prof. and companion animals, this parasite threatens Christina Strube3, Prof. Manuela Schnyder1 captive exotic and wild animals. Veterinary 1Institute of Parasitology, University of Zurich, diagnostics rely primarily on serologic Zürich, Switzerland, 2IDEXX Diavet AG, Baech, methods, most requiring species-specific Switzerland, 3Institute for Parasitology, Centre conjugates. Our laboratory has adapted for Infection Medicine, University of Veterinary the modified agglutination test (MAT) for Medicine, Hannover, Germany simultaneous IgG and IgM detection in warm- blooded animals, testing more than 18,000 Aelurostrongylus abstrusus is a worldwide samples over 30 years. The commercially occurring feline lungworm. The spectrum produced MAT is ideal in veterinary testing of clinical signs in infected cats ranges due to the nonspecific reagents suitable for from mild to severe respiratory distress. We testing multiple host species. However, the performed a seroepidemiological study to federally regulated reagents require special define prevalence and risk factors for A. import permits and the necessary tachyzoite abstrusus infections in Swiss cats and to

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 315 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

assess the biogeographic distribution of The family Cyclocoelidae comprises numerous this parasite. Sera of 4067 domestic cats flattened digenetic trematode species were tested for the presence of antibodies parasitizing in diverse organ systems and the against A. abstrusus by a novel ELISA and thoraco-abdominal cavity of birds. Most of the results correlated with biogeographic the cyclocoelid trematode species develop aspects. A subsample of 1000 datasets in wet land mollusks except Morishitium was used for risk factor analyses. Overall, polonicum, which is transmitted by terrestrial 10.7% (434/4067, 95% confidence intervals snails of genus Helicella. In 2018, 11 black [CI]: 9.7-11.7%) of the cats were tested birds (Turdus merula) from Northern Baden- positive, with variations from 0.0% to 20.0% Wuerttemberg were routinely examined for among ten different biogeographic regions. parasites by section after being found dead or Differences were significant between the severely injured and euthanized due to human Western (13.9%, CI: 11.4-16.7%) and the endpoints for health burden. Trematodes were Eastern (9.2%, CI: 8.0-10.5%) Swiss Plateau, found in 4 animals, being mainly located in possibly attributable to the suitability of the the thoraco-abdominal cavity above posterior areas for intermediate hosts. In total 90.3% air sacs, scattered from caudal part of lungs (392/434) of the seropositive cats originated until caudal end of thoraco-abdominal cavity from regions lower than 700 m above sea with a parasite burden range from 12 to 51 level. Correspondingly, 98.9% (429/434) of worms per bird. No trematodes could be positive samples were obtained from regions found in visceral organs (air sacs, lungs, liver with a mean temperature higher than -2 °C in or intestine). According to morphological January, suggesting altitude and temperature criteria (size and localization of testes, ovary, being limiting factors for A. abstrusus suckers, oesophagus etc.) and sequencing of infections. Concerning individual risk factors, 672 bp of 18S rRNA gene, the parasites were prevalence was higher in intact (15.5%, CI: identified as M. polonicum (Cyclocoelidae). 9.5-23.4%) than in neutered cats (5.8%, CI: 7.9- This species has been described in several 10.4%). Young adult cats (aged 11-22 months) European countries including Poland (Baltic were significantly more often seropositive seacoast), Czech Republic, Spain (Mallorca) (10/76, 13.2%, CI: 6.5-22.9%) than kittens aged and Italy, but no reports on occurrence of 1-10 months (1/34, 2.9%, CI: 0.1-15.3%) or adult M. polonicum are available from Southern and senior cats > 22 months (58/889, 6.5%, Germany and Northern Baden-Wuerttemberg. CI: 5-8.4%). We here confirm that the use of It can be assumed that M. polonicum was a serological test can contribute to improve introduced into this region by migratory birds. the identification of infected animals, through The spreading of the trematode parasite evaluation of risk factors on a population level might have been increased by Usutu virus, and for a better management on an individual which had caused massive Turdus merula level, overcoming the challenges represented die-off circulating in Southern Germany by faecal examinations and the correlated since 2011-2012, influencing the population underestimation of the occurrence of A. dynamics of black birds. The reduction of the abstrusus in cats. Further seroepidemiological black bird population in Germany could have studies are ongoing. led to additional immigration of M. polonicum infected black birds from neighboring countries. To our knowledge, this is the first detection of PS03.68 Morishitium Polonicum in M. polonicum in this region of Germany. Black Birds (Turdus Merula) in Baden- Wuerttemberg, Germany

Dr. Viktor Dyachenko1, Dr. Rüdiger Richter2, Dr. Ronald Schmaeschke3, Prof. Dr. Arwid Daugschies3, Dieter Barutzki1 1Tierärztliches Labor Freiburg GmbH, Freiburg, Germany, 2Veterinary Practice Cappel, Öhringen, Germany, 3Institute of Parasitology, Leipzig, Germany

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 316 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.69 Canine Cardio-Vascular in France and Dirofilaria remains apparently Helminthoses in Dogs in France: located to the southern part of the country, Distribution and Dynamics of Heartworm also possibly slowly extensive. However (Dirofilaria Immitis) and French-Heartworm the global increase of diagnosis of cases of (Angiostrongylus Vasorum) in France: Dirofilariosis suggest a wide distribution of Results From a National Survey With dogs as sources to mosquitoes vectors with Veterinary Clinics potential changes in the future.

Professor Patrick Bourdeau1, Cecile Roux1, Caroline Dumont1, Camille Douine1 PS03.70 Companion Animal Parasite Council: 1Unit DPM -Veterinary school - Oniris, Nantes, One Health in Practice France Mrs. Stella Self2, Dr. Michael Yabsley1, Dr. A national survey on canine Angiostrongylosis Robert Lund2, Dr. Christopher McMahan2, Dr. and Dirofilariosis was conducted in summer Christopher Carpenter3, Dr. Jenna Gettings1 2017 with questionnaires sent to veterinary 1University Of Georgia, Athens, United States, clinics, to evaluate the distribution and 2Clemson University, Clemson, United States, prevalence of these diseases covering the 3Companion Animal Parasite Council, Salem, period 2008-2017. Information obtained United States from 620 questionnaires was compared to previous studies (2000 and/or 2008). Over the past few decades, companion More than one case of angiostrongylosis animals have become an integral part of many and dirofilariosis was detected in clinics family households in the United States and from respectively 50 and 28 administrative around the world. Not only do pets share a territories (départements n = 90). Cases were home with their human counterparts, they considered autochthonous (= suspected to often accompany the family away from be acquired within the territory of activity of the home, particularly dogs. Consequently, the veterinary clinic) in 41 for Angiostrongylus companion animals and humans share in and 9 for Dirofilaria (6 in 2008). The two risks of exposure to a variety of vectors and parasites seem to overlap in only 6 areas, pathogens. The Companion Animal Parasite mainly located in the Mediterranean zone. Council (CAPC) recognizes this shared risk Up to 10 annual cases of Angiostrongylosis and aims to conduct scientifically rigorous were diagnosed in clinics from 8 areas. For studies on the relationship of infectious Dirofilaria, clinics diagnosing 2-4 annual disease between humans and animals. CAPC cases were found in 3 areas (up to 10 in currently maintains a repository of testing one). Angiostrongylus was widely distributed data from dogs and cats covering vector- but more frequently in 3 regions: Ile de borne pathogens (Borrelia burgdorferi, France (around Paris), Rhône Alpes (mid Anaplasma spp., Ehrlichia spp., and Dirofilaria east) and the historial region of South-west. immitis), gastrointestinal parasites (Toxocara Multiple cases in kennels were diagnosed spp., Ancylostoma spp., Trichuris spp., and in 19 areas. The distribution of Dirofilariasis Giardia spp.), and viral pathogens (Feline is much dispersed with more cases in Leukemia Virus, Feline Immunodeficiency the perimediteranean zone and Corsica. Virus). These data are used to maintain The dynamics was evaluated. In 19 areas surveillance of these pathogens within the Angiotrongylosis was considered increasing, United States through maps available at stable in 29. For Dirofilaria the diagnosed https://capcvet.org/maps which are updated cases (mainly considered imported) were monthly. Additionally, annual forecasts considered increasing in 24. Interestingly are provided in the early spring each Angiostrongylosis was diagnosed in 19 areas year predicting the seroprevalence of B. not previously identified (2000 and 2008) burgdorferi, Anaplasma spp., Ehrlichia spp., suggesting a possible extension. In return and D. immitis in dogs. A recent publication Dirofilariosis was newly mentioned only on describes the association between the canine area. Canine Angiostrongylosis is an active B. burgdorferi data and human incidence of and widely distributed canine helminthosis Lyme disease. This further substantiates the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 317 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

use of dogs as sentinels for zoonotic vector- PS03.72 A Rapid, Parasite-Dependent borne disease. CAPC aims to use canine data Cellular Response to Dirofilaria Immitis in the to inform on human risk of vector-borne Jird (Meriones Unguiculatus) disease with the ultimate goal of providing a tool for health care providers and public Christopher Evans1, Katherine Day1, Bridget health officials to use in the prevention and Garner1, Kaori Sakamoto1, Andrew Moorhead1 control of vector-borne disease. The data 1University Of Georgia, Athens, United States is available on a monthly and county-level scale, eliminating much of the missing data, The Mongolian jird (Meriones unguiculatus) underreporting, and reporting lags found in is long-recognized as a permissive host for human case reporting. With these, CAPC can the filarial parasite Brugia malayi. This is produce maps regularly that estimate the believed to result from the immunological level of risk of exposure to various vector- characteristics of the species, as particular borne pathogens. immunodeficient mouse models exhibit this same permissivity. At the same time, the jird is nonpermissive to another filarial parasite, PS03.71 The Prevalence and Intensity factors canine heartworm (Dirofilaria immitis), and so of Otodectic Mange In Cat In Two Districts of by elucidating differences in early response Pakistan (Lahore and Sheikhupura). to infection, we hope to identify mechanisms involved in the species-specific clearance of Uzma Mehreen1, Arfan Zaman1, Zohaib Shahid1, these parasites. We hypothesized that the Warda Qamar1, Fiaz Qamar1, Kazim Ali1 early clearance of D. immitis in intraperitoneal 1Cvas, Pakistan infection of the jird is immune-mediated and parasite species-dependent. The study was conducted to update the status of Otodectic mange in commensurate When D. immitis third-stage larvae (L3) were with age, sex, breed, housing and intensity. To injected intraperitoneally into jirds (200 each, this end 450 samples were collected from the n = 3), only 20% of parasites were recovered ear wax of cats brought on pets’ clinic from at 1h (range: 0 – 74), with 85% exhibiting district Sheikhupra and Lahore. Skin scraping host cell attachment. We then assessed the and ear wax were collected from all the Cats species-dependency of this cell attachment in selected for sampling and observed under vitro. Cultures were prepared as three groups: the Microscope. By statistical analysis the D. immitis L3, B. malayi L3, and co-culture of prevalence observed was 38.03% and 32.40% both parasites. Each group was cultured with in Lahore and Sheikhupura. The ubiquity in peritoneal exudate cells (PECs) from naïve cats with the age of less than 12 months and jirds and paired with a media-only control (20 more than 12 months was 39.67% and 29.8% L3 per well, n = 3 for 3 experiments). We then respectively. Prevalence rate was noted higher assessed host cell attachment and parasite in male cats (36.25%) than female (34.39%). survival microscopically after 20h incubation. No specific breed associations have been found. Outdoor cats have more prevalence In all conditions, cell attachment to D. immitis (46.85%) than those that lived indoor (12.29%) was 100%, while B. malayi was much lower So more the grubby environment more (mean = 5.56%), suggesting a strongly species- will be the prospects of ear mites. Besides dependent response from which B. malayi could this Cotton bud soaked in liquid paraffin is not confer immediate protection in co-culture. construct to be the easiest and the affordable When we replicated these experiments with method for the detection of ear mites. PECs derived from jirds subcutaneously infected Clean environment for housing and regular with B. malayi, results were similar (99.4% and ear cleaning is the best way to control its 4.72% of D. immitis and B. malayi, respectively, infestation. exhibited cell attachment). Applying Wright’s stain, the cells responsible for this L3 attachment were morphologically most consistent with lymphocytes, and the specific nature of this attachment is the subject of ongoing study.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 318 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.73 Diving Deeper into Fecal Counts: and correlating it with oocyst per gram Exploring New Technologies to Identify and (OPG) counts will create data needed Quantify Eimeria in Mixed-Species Samples by veterinarians, researchers, technical representatives, nutritionists and producers. Mr. Ryan Snyder1, Dr. Michele Guerin1, Dr. Billy Converting uninformative OPG counts Hargis2, Perryn Kruth1, Dr. Greg Page3, Dr. John into precise measures of parasite diversity Barta1 and relative abundance is prerequisite for 1University Of Guelph, Guelph, Canada, unraveling this disease complex and thereby 2University of Arkansas, Fayetteville, United improve control of coccidiosis in commercial States, 3Trouw Nutrition Canada, Guelph, poultry industries. Canada

Coccidiosis, caused by a number of protozoan PS03.74 MarkerDB: A Pipeline to Build parasites in the genus Eimeria, is a common Bespoke Databases of Marker Gene and costly intestinal disease affecting the Sequences for Identifying Taxa in Samples poultry industry globally. Eimeria species are ubiquitous in commercial poultry facilities, but Dr. Matthew Workentine1, Ms. Rebecca Chen1, presence does not equate with disease due to Ms. Jill De Rijke2, Ms. Nicolette Shaw2, Mr. the biological characteristics of the parasites’ Stefan Gavriliuc1, Dr. Libby Redman1, Dr. self-limiting life cycles. Multiple Eimeria species Russell Avramenko1, Dr. Janneke Wit1, Dr. can infect a chicken simultaneously with each Jocelyn Poissant1, Prof. John Gilleard1 parasite species varying in its pathogenicity 1Faculty of Veterinary Medicine, University and perceived commercial importance. of Calgary, Calgary, Canada, 2University of Therefore, identification of individual Eimeria Waterloo, Waterloo, Canada species in mixed samples is valuable for understanding this disease complex. Progress in next-generation sequencing technologies is revolutionizing our ability Molecular assays can provide a more to use molecular approaches to identify reliable means of identifying these parasites and quantify the presence of different based on DNA targets present in a sample. parasite and other pathogen species in Conventional PCR, both direct and nested, fecal, environmental and other samples. has been used successfully for Eimeria However, there are a variety of challenges species identification but standard PCR to the accurate identification of taxa, cannot provide quantification data. Newer particularly to the species level, including technologies may be able to provide the marker selection, bioinformatic pipeline necessary data for understanding relative choice and parameterization, reference abundance of Eimeria species in sample. sequence availability and database selection Digital droplet PCR (ddPCR) is a sensitive and curation. High quality databases are quantitative method that could be applied essential for accurately identifying taxa in to the identification and quantification of samples using marker genes such as the Eimeria species. A ddPCR-based method ribosomal small subunit (SSU) and the was developed that exploited mitochondrial internal transcribed spacer (ITS). Although genome targets that were genus-specific or large publicly available databases exist for species-specific. DNA and purified oocyst both the large and small ribosomal subunits, samples of known identities were tested in such as SILVA, there are few options for combinations to assess the sensitivity and other marker regions such as ITS-2 rDNA repeatability of this experimental assay. Early that may be more suitable for particular results have been promising using samples projects. Furthermore, large databases often of three Eimeria species considered to be the have long delays between release cycles. most important to industry with good assay markerDB is a pipeline to build databases of sensitivity. Following additional validation any chosen marker gene sequence for a given studies, samples from commercial facilities taxonomy built in R and snakemake. Currently with unknown contents will be tested. it supports ITS-2 and 18S rDNA sequences Obtaining relative species abundance but can be easily expanded to be use for any

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 319 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

gene. The pipeline retrieves sequences from analysis regarding Dirofilariosis. At the end NCBI annotated with the provided marker, of the study 22 out of 24 dogs remained identifies the correct region using a hidden clinically healthy with negative testing, giving markov model (using barrnap, https://github. a protection of 91.7% against Dirofilariosis. com/tseemann/barrnap) or covariance model Regarding leishmaniosis, the 4 dirofilariosis (using infernal) and formats them for common infected dogs as well as the car accident pipelines like RDP, dada2 and mothur. This were removed from the calculations; at pipeline provides speed and flexibility the end of the study, 22 of them remained beyond what is available in public databases. negative, giving an 88% protection against markerDB is available at https://github.com/ leishmaniosis. ucvm/markerDB. Those encouraging results show Effitix®spot- on can be one efficient option to PS03.75 Evaluation of the Preventive prevent Dirofilariosis and Leishmanisosis Efficacy of a Permethrin-Fipronil Based transmissions; results would need to be Spot-On (Effitix® Spot-On) for Canine confirmed on larger scale with a control Leishmnaniosis and Dirofilariosis in a Highly population. Endemic Area in Greece: An Open Field Trial

Dr. Elias Papadopoulos1, Dr. Olga Karteri2, Dr. PS03.76 Evolution of the Efficacy of Vanessa Chala3, Dr. Christelle Navarro3 a Combined Moxidectin-Levamisole 1Aristotle University of Thessaloniki, Greece, Treatment Against Resistant Gastrointestinal 2Virbac Greece, Greece, 3Virbac, Global Nematodes in Lambs Medical Department, France Lic. Sonia Luque1, Dr. Mercedes Lloberas2, Canine dirofilariosis (Dirofilaria immitis) and MV Patricia Cardozo2, Prof. Carlos Lanusse1, leishmaniosis ( Leishmania genus) are two Prof. Guillermo Virkel1, Lic. Cristina Farias1, MV vector borne diseases, occurring worldwide Paula Viviani1, Prof. Adrian Lifschitz1, Dr. Luis including now in northern countries(1)(2). Alvarez1 Therefore affordable and efficient options are 1Centro de Investigación Veterinaria de Tandil needed to struggle against reservoirs carriage, (CIVETAN), UNCPBA-CICPBA-CONICET, to protect both animals and humans health. Tandil, Argentina, 2Instituto Nacional de Thirty privately owned dogs, living outdoor, Tecnología Agropecuaria (INTA), Balcarce, in a highly endemic area of northern Greece Argentina (Serres), were recruited. In clinic diagnostic tests (Speed LeishK /Diro, Virbac BVT, La Nematodicidal combinations can be a valid Seyne sur Mer, FRANCE) were used to screen strategy to achieve effective nematode and exclude positive carriers (detection of anti control in the presence of drug resistance. leishmania’s kinesin antibodies and antigens The aim of the current trial was to evaluate from adult filaria) before enrollment. Each dog the pharmaco-parasitological outcome after was treated with Effitix® (Permethrin 44.88% the continuous use of moxidectin (MXD) and - Fipronil 6.01%) following manufacturer levamisole (LEV) as a combined treatment in recommendations, at inclusion time, then on lambs naturally parasitized with ivermectin- a monthly base, over 18 months (from June resistant gastrointestinal nematodes. Forty 2017 to November 2018). Dogs were monthly (40) lambs were divided into four groups blood sampled to follow their status. (n=10): untreated control and subcutaneously treated with either MXD (0.2 mg/kg), LEV Amoung the 30 dogs, one accidently died, (8 mg/kg) or with MXD+LEV (0.2 and 8 two were found to be dirofilariosis positive mg/kg, respectively). Blood samples were (in August and September respectively: collected at different times up to 1 (LEV) or which implied a contamination prior to 14 (MXD) days post-treatment. LEV and MXD enrollment), three additional leishmaniosis plasma concentrations were measured by infected dogs were removed from the study; HPLC. Faecal samples were collected on days all were excluded accordingly from the 0, 7, and 14 post-treatment to perform the

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 320 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

faecal egg count reduction tests (FECRT). were found in OP-resistant ticks. In total, No significant pharmacokinetic (PK) adverse the genomic DNA of 134 individuals from changes were observed for either MXD or LEV susceptible and resistant populations were after their co-administration in sheep. The analyzed. The I54V mutation was found in 133 clinical efficacy of the MXD+LEV combination individuals, the R86Q in 131 and the I48L in 44 was evaluated after its continuous use (3 individuals. In all ticks that survived ethion treatments/year) over five (5) years at the exposure, mutations I54V and R86Q were same farm. The initial anthelmintic efficacies detected. The I54V was found in only 20% of (1st year) were 99% (MXD), 85% (LEV) ethion-treated survivors. Both resistant and and 100% (MXD+LEV). Following repeated susceptible ticks presented any of these three annual treatments over five years, the clinical mutations, including ticks form a susceptible response for the combined treatment reached reference strain (100% with I54V/R86Q). The 87% efficacy. The combination reached results obtained in the present study disagree efficacies of 100% (1st year) and 98.5% with previous published data associating (5th year) against Haemonchus contortus. these mutations with OP resistance in CFT. Teladorsagia spp. and Trichostrongylus spp. Mutations in other acetylcholinesterase genes were the main nematode genera surviving the (BmAchE1 and 2) and metabolic detoxification individual and combined treatments. The co- may also contribute with OP resistance in this administration of MXD+LEV during five years tick species. resulted in a significant higher anthelmintic effect compared to MXD or LEV given alone. Even when MXD and LEV individual efficacies PS03.78 Anthelmintic Resistance and were reduced during the five-year period, the Common Worm Control Practices in Sheep combined treatment maintains acceptable Farms in Belgium efficacy levels against H. contortus. Prof. Edwin Claerebout1, Ms. Nathalie De Wilde1, Mrs. Eva Van Mael2, Mr. Stijn Casaert1, PS03.77 Polymorphisms in the Dr. Fiona Vande Velde1, Dr. Florian Roeber3, Acetylcholinesterase 3 Gene in Cattle Fever Ms. Pamela Vinueza Veloz1, Prof. Bruno Ticks (Rhipicephalus Microplus), Isolates Levecke1, Prof. Peter Geldhof1 from Uruguay and Southern Brazil 1Ghent University, Merelbeke, Belgium, 2Animal Healthcare Centre Flanders, Lier, Belgium, Br. Florencia Bariani1, Dr. Guilherme M. Klafke2, 3AusDiagnostics Pty, Beaconsfield, Australia Dr. Eleonor Castro-Janer1 1Udelar-school Of Veterinary, Montevideo, In contrast to many other European countries, Uruguay, 2Instituto de Pesquisas Veterinarias no data were available on the presence of Desiderio Finamor, Secretaria da Agricultura anthelmintic resistance in gastrointestinal do Estado do Rio Grande do Sul, Eldorado do nematodes in sheep in Belgium. A faecal Sul, Brazil egg count reduction test was performed in 26 sheep farms (29 flocks) in Flanders, The objective of this study was to Northern Belgium. Results indicated identify mutations in acetylcholinesterase widespread resistance against benzimidazoles 3 gene (BmAchE3) of cattle fever (albendazole, fenbendazole and ticks (CFT - Rhipicephalus microplus) mebendazole), with treatment failure (FECR < previously associated with resistance to 95 %) in all 8 flocks investigated. Haemonchus organophosphates (OP) in field isolates of contortus and Teladorsagia circumcincta CFT obtained in Uruguay and Rio Grande do were the predominant species after treatment Sul state, Southern Brazil. CFT populations failure. Amino acid substitutions associated (n=24) were submitted to the larval packet with benzimidazole resistance were detected test with ethion in order to characterize at the codon positions 167 (8%) and 200 phenotypic resistance. To identify nucleotide (92%) of the isotype-1 beta tubulin gene in polymorphisms in BmAchE3, we amplified H. contortus, codon positions 198 (47%) and and sequenced a segment of 308 bp where 200 (43%) in T. circumcincta and position 200 three mutations (I48L, I54V and R86Q) (100%) in T. colubriformis. Resistance against

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 321 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

macrocyclic lactones (ivermectin, doramectin formulations for oral administration in sheep and moxidectin) (FECR < 95%) was recorded were developed, employing a fluidized bed on 7 out of 20 flocks, mainly in H. contortus coating process adding different excipients and T. circumcincta. Treatment failure was in order to protect the fungus during its also observed for closantel (in combination transit through the gastrointestinal tract. The with mebendazole) (FECR 53%) and for aim of this work was to assess the storage monepantel (FECR 44%), on one farm each. stability of the fungal formulations developed. Trichostrongylus spp. and Cooperia curticei Two formulation prototypes (E3, E16) and were implicated with resistance against a control, consisting of the dried fungus monepantel. substrate, were storaged in glass vials at three temperatures (20 ± 2 °C, 25 ± 1 °C, 30 A questionnaire survey on farm management ± 3 °C) for 90 days. The response variables and worm control measures indicated that evaluated were the in vitro nematophagous worm control was often not sustainable. activity, fungal viability, fungal concentration Ewes and lambs were treated frequently (on and moisture content. The results showed average 2.6 and 3.2 times per year), mostly that the viability and concentration did without weighing. Only few sheep farmers not have significant variation (P<0.05) and (9%) regularly used faecal egg counts to remain higher than 1x106 UFC/g and 1 x107 monitor worm infections. Despite the FECRT UFC/g respectively, the moisture content also showing treatment failure, most of the farmers remain under 5 %. The ability of the fungus perceived the efficacy of anthelmintics as to trap the nematodes was higher than 80 good (54%) to very good (29%). Only 12% % during the storage period which is an and 4% evaluated the anthelmintic efficacy as important charasteristic to continue with the mediocre or insufficient, respectively. product development. Agrosavia will test the In conclusion, anthelmintic resistance is developed formulations in sheep at different widespread in Belgian sheep flocks. There climatic conditions. is an urgent need to efficiently promote sustainable worm control practices to sheep farmers and veterinarians. PS03.80 Canine Heartworm Treatment Using a Combination of ProHeart® SR 12 (Injectable Moxidectin) and Oral Doxycycline PS03.79 Storage Stability Study of Solid Based Formulations of the Nematophagous Fungus Bruno Alberigi1, Julio Fernandes1, Jonimar Duddingtonia Flagrans for Biological Control Paiva1, Flavya Mendes-de-Almeida2, Fabiana of Gastrointestinal Nematodes in Sheep Batalha3, Alexandre Merlo4, Norma Labarthe5 1Universidade Federal Rural Do Rio De Janeiro, Dr. Diego Cortes-Rojas1, Elizabeth Cespedes- Seropédica, Brazil, 2Universidade Federal Gutierrez1, Jaime Andres Cubides- Cardenas1, Fluminense, Niterói, Brazil, 3Universidade do MarthaGomez-Alvarez1 Grande Rio, Rio de Janeiro, Brazil, 4Zoetis, São 1Agrosavia, Mosquera, Colombia Paulo, Brazil, 5Fundação Oswaldo Cruz, Rio de Janeiro, Brazil The inappropriate use of chemical antiparasitic agents in ruminants has Dogs infected with Dirofilaria immitis may generated anthelmintic resistance in develop a life-threatening disease affecting gastrointestinal nematodes. Biological the heart and lungs. Treatment should focus control using the nematophagous fungus is on the elimination of the adult worms as a promising alternative due to its capacity to soon as possible after diagnosis of infection, develop traps that capture the nematodes as this will minimize side effects from killing in free living phase reducing the use of of the worms. ProHeart® SR-12 (SR 12) is chemical drugs. A native species of the approved in Australia, Japan and some fungus Duddingtonia flagrans (BGMSABV- Latin American countries for the prevention Df-Col-H-001-2014) was isolated in Agrosavia of heartworm disease for 12 months via a Colombia and a fermentation medium was single subcutaneous injection. In this study, developed for its production. Solid- based conducted in Brazil, after obtaining owner

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 322 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

consent 16 naturally infected dogs were given both separately and co-administered confirmed positive for microfilariae and adult to calves in two commercial farms (A and heartworm antigen (SNAP 4Dx Plus). Dogs B). Sixty(60) male calves naturally infected were then administered SR-12 (0.5 mg/kg of with gastrointestinal nematodes were injectable sustained release moxidectin) at the randomly allocated into four groups(n= 15): same time as they started daily administration Control: animals did not receive anthelmintic of doxycycline (10 mg/kg BID) for 30 days treatment; IVM: treated with IVM (0.2 mg/ (Day 0). All dogs received the same dosing kg); LEV: treated with LEV (8 mg/kg); schedule of both drugs every 6 months IVM+LEV: simultaneously treated with IVM (modified from the approved 12 month dosing and LEV (at the same dose rates). Seven (7) interval for SR-12 for heartworm prevention) animals from each treated group (Farm A) until two negative adult heartworm antigen were randomly selected to perform the PK tests (SNAP 4Dx Plus) were obtained study. Drug concentrations were measured 6 months apart. Dogs had chest x-rays, by HPLC. The efficacy was determined at 14 echodopplercardiograms, Knott´s tests days after treatment by the FECRT. The IVM and antigen testing done every 6 months. area under the concentration vs time curve Microfilariae counts were performed on Days (AUC) obtained after administration of IVM 0, 30 and 150. On Day 150, all dogs were alone (274±65.1ng.d/mL) was similar to that amicrofilaremic. Eleven dogs became antigen obtained after IVM co-administered with LEV negative at 6 months after the first dose and 5 (295±111 ng.d/mL). Likewise, LEV AUC values dogs at 12 months of treatment. At 12 months were similar after LEV administration alone of treatment 7 dogs were considered free of (8.90±2.69 g.h/mL) or combined with IVM the infection and by July 2019, as the study (9.11±1.82 g.h/mL). No adverse PK interactions is ongoing, 4 additional dogs will be tested were observedµ after the combined treatment, to confirm their infection status at 12 months with similarµ PK parameters (P>0.05) obtained and the 5 remaining dogs at 18 months of between the single-drug and combination- treatment. All dogs tested 6 months after the based strategy. In Farm A, the overall first negative antigen test remained negative efficacies were 54%(IVM), 99%(LEV) and 6 months later. Preliminary disease evaluation 100%(IVM+LEV). While Cooperia spp. survived suggests that lung and heart lesions tended to IVM treatment, Ostertagia spp. survived remain unchanged throughout the treatment. LEV treatment. In fact, the efficacy against Cooperia spp. was 41%(IVM), 100%(LEV) and 100%(IVM+LEV), and the efficacy against PS03.81 Combined Use of Ivermectin and Ostertagia spp. was 91%(LEV), 100%(IVM) Levamisole to Control Resistant Nematodes and 100%(IVM+LEV). Similarly, in Farm B, in Cattle: Assessment of Pharmacokinectic total efficacies were 55%(IVM), 99%(LEV) and Interactions and Therapeutic Responses 100%(IVM+LEV). Although LEV alone achieved high efficacy in both farms, the combination Dr. Candela Canton1, Dr. Laura Ceballos1, was the only treatment that achieved Dr. Laura Moreno1, Veterinarian María Paula 100% efficacy against all genera (Cooperia, Domínguez1, Ms. Lucila Canton1, Dr. Carlos Ostertagia and Haemonchus). Further work Lanusse1, Dr. Luis Ignacio Alvarez1 is required to understand the advantages 1Laboratorio de Farmacología, Centro de of nematodicidal combinations in different Investigación Veterinaria de Tandil (CIVETAN), commercial cattle farms. UNCPBA-CONICET-CICPBA, Tandil, Argentina

Combination of anthelmintics with different mechanisms of action has been suggested as an effective strategy to delay the development of resistance. In this context, the current study evaluated the efficacy and the potential pharmacokinetic (PK) interactions occurring after the subcutaneous administration of ivermectin (IVM) and levamisole (LEV)

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 323 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

PS03.82 Impact Evaluation of a Regular This trial allows to confirm the persisting Shampoo Use, on the Efficacy of a killing effect of the permethrin-fipronil Permethrin and Fipronil Combination (Effitix® combination, even when applied just after Spot-On), Against Flea Infestations in Dogs: shampooing, over 35 days together with A Randomized, Controlled Study a weekly bathing of the dog. Those very encouraging results would need to be Dr. Fabio Barbour-Scott1, Dr. Thais Correia confirmed on a larger scale. Azevedo1, Dr. Debora Azevedo Borges1, Dr. Difrey Ribeiro Campos1, Dr. Priscilla Cardim de Oliveira1, Dr. Marcia Christina Alves Teixeira2, PS03.83 Efficacy of Topical 0.5% w/V Dr. Vanessa Chala3, Dr. Stephane Bonneau4 Eprinomectin (EPRINEX®) Administered at 1Universidade Federal Rural do Rio de Janeiro, 1 Mg per Kg Body Weight Against Larval Seropédica, Rio de Janeiro, Brasil, 2Virbac Oestrus Ovis Infestation in Sheep and Goats Brasil, R and D Department, São Paulo, Brasil, 3Virbac, Global Medical Department, Carros, Steffen Rehbein1, Elias Papadopoulos2, France, 4Virbac, R and D Department, Carros, Zvezdelina Kirkova3, Konstantinos France Arsenopoulos2, Petar Iliev3, Renate Rauh1, Hailun Wang4, Becky Fankhauser4 Regular bathing can be required in dogs, 1Boehringer Ingelheim Vetmedica GmbH, especially in subjects with skin conditions Kathrinenhof Research Center, Rohrdorf, impairment, which can discourage from spot- Germany, 2Aristotle University, School of on use, especially when ingredients have no Veterinary Medicine, Thessaloniki, Greece, or few systemic activity. 3Trakia University, Department of Parasitology, Stara Zagora, Bulgaria, 4Boehringer Ingelheim This study evaluates the impact of a weekly Animal Health USA, Inc., Duluth, GA, United neutral shampoo (Shampoo Pet Bril ®Neutro) States application on the efficacy of a Permethrin (44.88%) plus Fipronil (6.01%) based spot- INTRODUCTION: Oestrus ovis larvae mainly on (Effitix®, Virbac, Brasil), regarding fleas parasitize sheep and goats, but humans are infestations, in adult dogs. Eighteen healthy also vulnerable to infestation. Currently, there Beagle dogs were equally randomly allocated are no products authorized in Europe for to group 1 (Effitix® without shampoo), the control of oestrosis for use in lactating group 2 (Effitix® with weekly shampoo) or dairy animals. Topical 0.5% w/v eprinomectin, control group (no treatment). Treatment was administered at 1 mL per 5 kg body weight applied once on D0, following manufacturer to sheep and goats, has been recently recommendations (at least 60mg/kg of authorized in Europe as EPRINEX® Multi as permethrin and 6.7mg/kg for Fipronil). Dogs broad-spectrum anthelmintic with zero hours were infested with +/-100 fleas on D0 (after milk withdrawal and was thus evaluated treatment), D5, D12, D19, D26, D33, D40, against O. ovis larval infestation. D47, and D54. Flea counts were performed regularly 48 hours after each flea challenges. METHODOLOGY: Three blinded clinical Dogs were subsequently bathed with 50 ml of studies compliant with GCP and WAAVP shampoo and dried with a fluffy towel on D0 guidelines were conducted in Bulgaria (sheep) (before treatment application for group 1 and and Greece (sheep and goats). At each 2) and on days 8, 15, 22, 29, 36, 43 and 50. location, recovery of O. ovis larvae from five of six or six animals necropsied pre-study The mean geometrical efficacies against fleas’ supported the inclusion of animals into the infestation (prevention) were equal to 100% studies (34 adult female sheep or goats for both treated groups as soon as D2, until per study). Animals were ranked based on D21. Subsequent assessments on D28, D35, pre-treatment bodyweight and allocated at D42 allowed the calculation of a high efficacy random to remain untreated (control) or to be (100% vs 99.2%, 98.92 vs 97.97%, and 98.06 vs treated with EPRINEX® topically. Three weeks 94.74% for group 1 and 2 respectively). after treatment, animals were necropsied and O. ovis larval counts were established.

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 324 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

RESULTS: Treatment was well accepted The goal of this study was to determine and no health problems were observed the efficacy of the first moxidectin (MOX) throughout the studies. Live O. ovis larvae treatment and MOX-OBZ combination were recovered from 13 of 17 and 16 of 17 treatments against this cyathostomin control animals in the sheep studies and from population, and observe changes in OBZ all 17 controls in the goat study comprising efficacy. Treatments were given when ten first, second and third instars in each study. ponies exceeded 100 eggs per gram. Two In each study, EPRINEX®-treated animals had OBZ treatments (initial and final), one MOX significantly fewer live O. ovis larvae than single-active, two combination treatments the controls (p<0.001). Efficacy of EPRINEX® were given over the course of two-and-a-half treatment, based on O. ovis larval counts years. Fecal egg counts were performed every of treated vs. untreated animals, was 100% two weeks, and efficacies were evaluated and 91.3% against infestation with all three using the fecal egg count reduction test. The O. ovis larval stages in sheep and in goats, absolute efficacy of MOX single-active also respectively. provided 100% efficacy for both combination treatments. The egg reappearance period In conclusion, results of the studies for MOX was 16 weeks, and the combination demonstrated EPRINEX® to be an efficacious treatments were 12 and 18 weeks. There was treatment against ovine and caprine oestrosis no significant difference between the first and thus extend the efficacy profile of this and last OBZ treatment efficacies. The was product beyond nematode endoparasites. no evidence of cross-resistance to MOX from the previous OBZ/PYR resistance status. Combining OBZ with MOX did not PS03.84 Can Combination Deworming affect the efficacy as MOX single-active Conquer Double-Drug Resistance? Moxidectin was already 100% efficacious. The lack of and Oxibendazole Combination Treatments improvement in OBZ efficacy suggests that Against Double-Drug Resistant Cyathostomins combination treatments using an active where Naïve to Macrocyclic Lactones resistance is heavily established may not provide a beneficial alternative regimen for Jessica Scare1, Dr. Eugene Lyons1, Dr Ashley cyathostomin control. Longer term studies Steuer1, Brittany Jones2, Morgan Clark2, Dr and increasing the refugia may affect the OBZ Martin Nielsen1 efficacy and warrants future investigation in 1University of Kentucky, Gluck Equine this population. Research Center, Lexington, United States, 2Lincoln Memorial University, College of Veterinary Medicine, Harrogate, United States PS03.85 In Vitro Characterization of the Effects of Polyhexamethylene Biguanide Equine cyathostomins are well-known (PHMB) Alone or in Complex With TLR4 contributors to the anthelmintic resistance or TLR9 Agonists in Leishmania Infantum crisis with wide-spread resistance to the Infected Canine Macrophages benzimidazole drug class, and reports of resistance to the pyrantel (PYR) Dr. Pamela Martinez-Orellana1, Dr. Klaudia and macrocyclic lactone drug classes. Kloc-Muniak2, Miss Marta Baxarias1, Dr. Liam Combination deworming is suggested as an Good2, Dr. Laia Solano-Gallego1 alternative control regimen for drug-resistant 1Dep. Medicina I Cirurgia Animals, Universitat parasites and is commonly implemented Autonoma de Barcelona, Bellaterra, Spain, in the small ruminant community. Some 2Department of Pathobiology and Population products exist for horses despite the lack of Sciences, The Royal Veterinary College, supporting evidence. Previously, we found University of London, Potters Bar, United that the combination of oxibendazole (OBZ) Kingdom and PYR did not effectively control double- drug resistant cyathostomins. This population The dog is the main reservoir for Leishmania remains naïve to macrocyclic lactone drugs infantum infection which can manifest as and offers a unique opportunity for research. subclinical, self-limiting or a severe and fatal

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 325 63rd Annual Livestock Insect Workers Conference POSTER SESSIONS

disease. Limited drug treatment options are available. New drugs as well as new immunomodulators for treatment of canine and human leishmaniosis are actively pursued. Polyhexamethylene biguanide (PHMB) is a polymer that has broad antimicrobial spectrum and is used as a disinfectant and antiseptic. However, very limited studies are available regarding PHMB’s activity against protozoan parasites. Here, we examined how PHMB alone or in complex with Toll like receptor agonists (TLRa) selectively kills L. infantum parasites. DH82 cells were infected at a 10:1 parasite:host cell ratio with L. infantum promastigotes. After 3 hours, the cells were washed and treated with 1 g/ mL of PHMB alone or combined with TLR4a (Monophosphoryl Lipid A) or TLR9a (CpGµ ODNs) agonists. At 24h, cells were fixed with methanol and stained with diff quick to determine the rate of intracellular infection. Furthermore, the supernatants were collected for measurement of IL-6 and TNF- by ELISA. Results indicated significantly lower infection when cells were treated with PHMBα alone (p=0.043), PHMB+TLR4a (p=0.0031) and PHMB+TLR9a (p=0.0338) compared with infected but not treated cells. Additionally, higher production of TNF- was observed after treatment with PHMB alone or in combination with TLR4a whenα compared with uninfected and infected untreated- macrophages (p=0.043), but not with TLR9a. Moreover, PHMB+TLR4a induced significantly higher production of TNF- when compared with PHMB+TLR9a (p=0.043). No differences were found in IL-6 concentrationsα in all conditions studied. In conclusion, PHMB combined with TLR4 agonists is a potent antiparasitic drug combination and also induces a proinflammatory response, as demonstrated by decreased infection and increased TNF- production.

α

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 326 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 327 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Author Index

A Alberigi, Bruno...... PS01.64, OA23.04, PS03.80 Alberti-Navarro, Aldo...... PS03.14 Abbas, Haider...... OA17.06, OA50.03, PS03.48 Alcala-Canto, Yazmin...... PS01.11 Abbas, Rao Zahid...... OA09.07, OA28.04, Alcalá-Canto, Yazmín...... PS03.14 ...... OA32.05, OA49.05 Alencar, Luciana Magalhães Rebelo...... PS01.25 Abdalla, Adibe...... PS01.08, PS02.14, PS01.71 Alencar , Luciana M. R...... PS01.26 Abdalla, Muna Ali...... PS02.13 Alexandre, Fanny...... PS02.34 Abdelsamad, Noor...... OA06.04 Ali, Kazim.PS01.85, PS02.56, PS03.71, OA50.07 Abdel-Shafy, Sobhy...... OA08.08 Ali, Qasim...... PS01.77, OA46.06 Abdoli, Alireza...... OA32.03 Allen, Kelly...... OA11.01, OA13.03, OA26.03, Abdu, Amira...... OA47.04 ...... PS03.10, PS02.01 Abdul latif, Asma...... OA35.03 Allenspach, Karin...... PL02.02 Abdullah, Mohd Tajuddin...... OA40.01 Almeida, Caroline...... PS01.64 Abdullah, Swaid...... OA44.05 Al-Mohammed, Hamdan...... OA08.08 Abongwa, Melanie....PS01.15, PS01.21, OA01.06, Alnassan, Alaa...... PS02.51 ...... OA19.04 Alnassan, Alaa Aldin...... OA26.02 Abraham, Ambily...... OA28.02, OA50.02 Alonso-Díaz, Miguel Ángel.....PS01.28, OA50.01 Abrahao, Carolina...... OA38.08 Alota, Sherwin...... OA39.02 Abruzzo, Nicole...... OA06.04 Alozor, Ijeoma Angela...... OA15.04 Abu El Ezz, Nadia...... OA42.01 Alvarez, Luis...... PS01.78, PS02.83, PS03.76, Acker, Jason...... OA40.06 ...... PS03.81, OA31.05, OA35.01, OA39.08 Ackermann, Rebecca...... PS01.42 Alvarez Rojas, C.A...... OA16.01 Adam, Babagana Mohammed...... OA15.04 Alves, Leucio...... OA23.03 Adam, Brian...... PS02.06 Alves, Luis...... PS01.30 Adams, Amanda...... OA33.04 Alves, Maria Luana...... OA18.06 Adamu, Mathew...... PS03.16 Alves Teixeira, Márcia Cristina...... PS03.02, Adeleke, Matthew...... PS02.35 ...... OA39.05 Adjou, Karim...... OA25.04, OA25.05 Alyousif, Mohamed...... OA08.08 Adorno, Oswaldo José Christe...... PS02.50 Alzan, Heba...... OA17.04 Agarwal, Manish...... PS03.36 Amadesi, Alessandra...... OA12.01, PS03.07 AGUERRE, SOPHIE...... PS01.09 Amairia, Safa...... PS01.33 Agwa, Akello...... OA50.06 Amarante, Alessandro...... PS01.08, PS01.26, Ahmed, Abdalla Fadlalla Azrug...... PS01.29 ...... PS02.14, OA07.06 Ahmed, Alaa...... OA37.01 Ambrosini, Yoko...... PL02.02 Ain-Baaziz, Hacina...... OA25.05 Amdouni, Yosra...... PS01.33 Airs, Paul...... PS03.21, PS03.35 Amery-Gale, Jemima...... PS02.67 Akanji, Bukunmi...... OA01.06 Ammar, Sawsan,...... OA30.01 Akashi, Karen...... PS02.23, PS02.27 Amodie, Deborah...... OA13.03 Akashi, Karen T...... PS03.50 Andersen, Erik...... OA19.01 Akashi, Karen Tieme...... PS01.48, PS02.45, Anderson, Bryan...... PS01.69 ...... PS02.84, PS03.28 Anderson, David...... OA49.01 Akira Nakamura, Alex...... PS01.38 Anderson, Haley...... PS02.25, OA33.03 Akram, Muhammad Zeeshan...... OA21.07 Anderson, Michael...... PS01.69 Akyol, Volkan...... PS02.16 Andrade, Mariana Fogale...... PS02.73, PS03.51 Al Khoury, Charbel...... PS02.10, OA42.02 Andrade, Wagner...... OA23.03 Al-Adhami, Batol...... OA25.07 André, Nicole M...... OA43.01 Alanazi, Abdullah...... OA08.08 Andreotti, Renato...... PS01.46 Alasmari, Saeed...... PS03.42 Andrew, Nigel R...... OA14.03 Alatta, Abdelaziz Atta M...... PS01.29 Andrews, Leigh...... OA36.04 Alberdi, Pilar...... OA27.02 Ang, Ching-Seng...... OA19.06

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 328 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Angelou, Athanasios...... PS01.80, PS03.63 Bah, Saidal Ali...... PS02.16 Angerer, Jay...... OA49.03 Bai, Meng-Jie...... OA04.06 Angulo-Tisoc, José...... PS01.50 Baker, Kacie...... PS03.39 Anlind, Alice...... PS02.07 Bakshi, Mariam...... OA07.05, OA24.01 Annoscia, Giada...... OA27.03, OA27.05 Bal, Mandeep Singh...... PS01.58 Antunes, Joaquim...... OA38.08 Balaña, Rafael...... OA36.08 Antunes, Paola...... OA39.01 Balbino, Daniele Rizatto...... PS01.74, PS01.22, Anwer, Azra...... OA50.05 ...... PS02.23, PS02.73, PS03.28, PS03.56 Aoian, Raffi...... OA22.03 Baldwin, Luke...... OA32.01 Aono, Fernando...... PS01.02 Ballent, Mariana...... PS01.78, PS02.83 Apple, Hadley...... OA21.05 Ballesteros, Cristina...... OA01.04 Araújo, Shirley Batista da Silva...... PS01.57, Baltrusis, Paulius...... OA24.02 PS02.50, PS03.65 Balogun, Emmanuel...... OA29.02 Araújo Fontenele, Janice...... PS02.63 Baneth, Gad...... PL03.02, OA21.04 Araújo Frota, Gracielle...... PS01.70 Bangoura, Berit...... OA46.01 Araújo Pacheco, Poliana...... PS02.44 Baraibar, Florencia...... PS01.75 Araúz Viol, Milena...... PS01.38 Baraibar, Magdalena...... PS01.75 Arcila-Fuentes, William...... PS01.73 Barash, Nanelle R...... OA05.04 Arfuso, Francesca...... OA21.01 Barbosa, Isabela...... PS03.64 Armstrong, Brittany...... PS02.70 Barbour-Scott, Fabio...... PS03.02, PS03.18, Armstrong, Rob...... OA01.03 ...... PS03.82, OA39.05 Arocho Rosario, Charluz...... OA49.04 Bariani, Florencia...... PS03.77 Aroian, Raffi...... OA28.02, OA50.02 Barker, Day...... OA33.04 Arranz, Juan José...... OA41.05 Barnett, Craig...... PS03.11, OA43.03 Arsenopoulos, Konstantinos.PS01.80, PS03.63, Barone, Carly...... OA12.05, PS03.23 PS03.83 Barret, Kayleigh F...... OA04.01 Artemiou, Elpida...... PS02.58 Barret, Lynn K...... OA04.01 Ashraf, Kamran...... OA46.06 Barrett, Annie...... PS02.01 Aslam, Bilal...... OA49.05 Barros, Jacqueline...... PS01.46 Au, Vinci...... PS01.72 Barros, Luciano Antunes...... PS02.84 Aubrey, Marissa...... OA36.07 Barros, ATM...... OA34.02 Audhuy, Stéphane...... PS01.60 Barry, Bridget...... OA47.03 Auger, Clément...... OA13.04 Barta, John...... PS01.41, PS03.41, PS03.73, Austin, C.J...... PS03.13 ...... OA37.05, OA37.06 Austin, Corrine...... OA33.06 Barta, John R...... PS01.53, PS02.67 Autheman, Delphine...... OA29.01 Barta , John R...... OA14.05 Avelar, B...... PS02.82 Bartholomay, Lyric...... OA11.05, OA44.04 Avenant-Oldewage, Annemarie...... PS01.62 Bartley, David...... PS01.79, PS03.24, Avramenko, Russell...... PL02.04, PS03.31, OA19.02, OA19.03, PL02.04, OA26.05, PS03.26, PS03.74, PS01.17, ...... OA24.03, OA24.04, OA31.01, OA36.04 OA19.02, OA24.06, OA41.01 Barutzki, Dieter...... PS03.68 Avula, Jacob...... PS02.20, PS03.22 Bąska, Piotr...... PS02.66 Awadi, Sofia...... PS01.33 Bassil, Mila...... OA40.06 Awouafack, Maurice...... PS03.16 Basto-Estrella, Gertrudis...... PS01.73 Azevedo, Bianca...... PS01.74, PS02.23, PS02.73, Batalha, Fabiana...... PS03.80 PS03.51 Baumgartner, Katrin...... PS02.51 Azevedo, T...... PS01.54 Baxarias, Marta...... PS03.85 Azevedo Borges, Debora...... PS03.18, PS03.82 BC, Parthasarthi...... OA49.07 Beall, Melissa...... OA05.01, OA05.05, OA10.02, B ...... OA26.01 Beall, Melissa...... OA08.07 Backwell, Amber Lynn...... OA48.07 Beam, Rachel...... PS02.01 Bader, Jonatan...... PS02.03 Beasley, Anne...... OA15.02 Beck, Paul...... OA36.03

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 329 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Becker, Marcelo...... OA39.01 Bornancin, Louis...... OA28.01, OA28.03 Beesley, Nicola...... OA24.07 Borrás, Pablo...... PS02.54, PS02.53, OA21.04 Beineke, Andreas...... OA40.02 Bosco, Antonio...... OA12.01, PS02.81, OA20.03, Bekelaar, Kiliana.....PS03.34, OA41.03, OA07.03 ...... PS03.07 Belem, Adrien...... PS03.47 Bossio, Felipe...... PS02.78 Bellaw, Jennifer...... OA38.05, OA48.07 BOUCHEZ, Caroline...... PS01.01 Bellier, Sylvain...... PS01.03 Bouhsira, Emilie...... OA13.06, PS03.58 Bencosme Cuevas, Emily...... OA08.01 Boulajfen, Henda...... OA27.06 Benedetti, Laura...... OA14.04 Bouman, Rick...... OA27.01 Benesi, Fernando José...... PS03.62 Bourdeau, Patrick. PS01.65, PS02.34, PS03.69, Bennett, Carolyne...... PS01.69 ...... PL03.03 Benoit, Joshua...... OA43.02 Bourguinat, Catherine...... OA01.04 Berger, Audric...... OA13.06 Bousquet, Eric...... PS03.13 Berger, Laura...... OA27.01 Bowles, Joy...... PS02.19, PS03.20 Bergström, Thomas F...... OA48.03, OA48.04 Bowman, Dwight..... PS01.34, PS01.61, OA01.02, Berke, Olaf...... OA20.02 OA05.01, OA10.01, OA21.06, OA15.05, OA47.03 Berkebile, Dennis...... PS02.33 Bown, Kevin...... OA15.04 Bernardo, John Michael...... OA39.02 Braff, Jennifer...... OA05.01, OA05.05, OA26.01 Bernigaud, Charlotte...... OA13.05, PS02.10 Braff, Jennifer...... OA08.07 Berriman, Matthew...... OA24.04, PS03.24 Brandao, Leonardo...... OA23.03 Bertocchi, Marie...... PS01.76 Brandson, Erika...... PS01.17 Beugnet, Frederic.PS01.43, OA08.03, OA13.01, Branigan, Marsha...... PS03.32 ...... OA13.05, OA27.05, PS02.40, OA27.03 Brayton, Kelly...... PS01.42 Bevelock, Thomas...... OA21.03 Brazis, Pilar...... OA23.06 Bhushan, Chandra...... OA39.03 Bredtmann, Christina...... OA12.04 Bier, Nadja...... OA30.04 Breitschwerdt, Edward B...... OA05.04 Biguezoton, Abel...... PS03.47 Bresciani, Katia...... OA09.03, OA37.03 Birgel Jr., Eduardo Harry...... PS03.62 Brewer, Matthew .....OA16.05, PS01.15, PS03.05, Birkenheuer, Adam J...... OA05.04 ...... PS03.27 Birtles, Richard J...... OA15.04 Briand, Amaury...... OA13.05 Blaga, Radu...... PS01.68 Briant, Emmanuel.... PS01.01, PS01.80, PS03.02, Blagburn, Byron...... PS01.39, PS01.61, OA06.03, ...... OA39.05 ...... OA06.05, PS02.19, PS03.20 Brianti, Emanuele....OA18.05, OA21.01, PS03.29 Blake, Damer...... PS02.35 Brinker, Emily...... PS01.39 Blasi, Dale...... PS02.06 Brisseau, Nadine...... OA02.04, OA02.05 Blatchford, Richard...... OA32.03 Brito, Danilo Rodrigues Barros...... PS01.16 Bodhipongsathorn, Thiantada...... PS02.36 Britti, Domenico...... PS02.81 Boersma, Melissa...... OA06.03 Britton, Collette...... OA24.03 Bogema, Daniel...... OA17.03 Bron, Gebbiena...... OA11.05, PS02.18, OA44.04 Boleaga-Rivera, Lidia...... PS01.05 Brorsen, Wade...... PS02.06 Bompadre, Thiago F V...... PS02.14 Brosnan, Sorcha...... OA49.06 Bonatte Junior, Paulino...... PS01.46 Brown, D. Andrew...... OA01.01 Boncea, Ana Maria...... OA21.02 Brown, Grant...... PS01.69 Bonde, Charlotte Smith...... OA28.01 Brown, Justin...... OA09.02, OA30.01 Bonilla, Denise...... OA03.04, PS03.59, OA49.02 Browne, Colm...... PS02.59 Bonneau, Stephane...... PS03.18, PS03.82 Browne, John...... OA07.02, OA49.06 Bonneau, Stephane...... PS01.80 Brummett, Marissa...... PS03.23 Bonnet, Sarah...... OA08.05 Bryant, Rachael...... OA02.02 Bordes, Lea...... OA46.02, PS02.74 Buch, Jesse...... PS01.69, OA05.01, OA05.02, Borel, Nicole...... OA06.07 ...... OA05.05, OA26.01 Borges, Dyego...... PS03.05 Buch, Jesse...... OA08.07 Borges, Fernando...... PS01.02, PS03.05 Buffarini, Miguel...... OA31.05 Borges Argáez, Rocío de Lourdes...... PS01.27 Bui, Khanh Linh...... OA27.03, OA27.05 Borkowski, Emma...... PS02.20, PS03.22 Bull, Katie...... PS01.79

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 330 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Buono, Francesco.OA05.05, OA26.01, OA33.01 Castro Janer, Eleonor...... PS02.32 Burgio, Federica...... OA01.05 Castro-Janer, Eleonor...... PS01.75, PS02.31, Burkman, Erica...... PS01.07 ...... PS03.40, PS03.77 Burnett, Peter...... OA06.08 Cavalera, Maria Alfonsa...... OA21.01 Bush, Russell...... OA30.03 Cavichiolo, José Roberto...... PS03.62 Buzetti, Wilma...... OA18.06 Ceballos, Laura...... PS01.78, PS03.57, PS03.81, Buzzulini, Carolina...... PS02.52 ...... OA31.05, OA35.01 Cerna, Corpus...... PS01.51 C Cespedes, Javier...... PS02.54 Céspedes-Gutierrez, Elizabeth...... OA50.03, C., Ansar Kamran...... OA42.04 ...... PS02.75, PS03.79 C., Shanmugnath...... OA49.07 Cestari Pierucci, Julia...... PS01.38 Cabezas-Cruz, Alejandro...... OA27.02 Cevallos, William...... OA45.04 Cabral, Welton...... PS03.05 Chagas, A.C.S...... PS02.41 Cabrera, María...... PS01.50, PS01.51 Chagas, Ana...... PS02.22 Cabrera Rubio, Raul...... OA38.07 Chagas , Isabella...... PS01.59 Cadenazzi, Gabriela...... PS01.82 Chaimee, Thitirat...... PS01.49 Cai, Xuepeng*...... PS02.17 Chala, Vanessa...... PS01.01, PS01.34, PS01.60, Cain, Jennifer...... OA15.01, OA33.02 PS03.02, PS03.08, PS03.18, PS03.63, PS03.75, Calvani, Nichola...... OA15.02, OA30.03 ...... PS03.82, OA39.05 Calvopiña, Manuel...... OA45.04 Chalut, Bailey...... PS03.23 Camara Alves, Leucio...... OA20.03 Chambers, Alex...... OA09.05 Campbell, John...... PS02.76, OA36.02 Chan, John...... OA22.07 Campestrini, Luciano...... OA50.04 Chandrashekar, Ramaswamy...... PS01.66, Cançado, PHD...... OA34.02 ...... PS01.69, OA05.01, OA05.03, OA05.05, Candy, Paul...... OA09.05 ...... OA26.01, OA27.04 Cantacessi, Cinzia...... OA43.06 Chandre, Fabrice...... OA13.06 Canton, Candela.....PS02.83, PS03.81, OA31.05, Chaparro-Gutiérrez, Jenny J.. PS01.67, PS02.78 ...... OA35.01, OA39.08 Chapman, Aly...... PS03.66 Canton, Lucila...... PS03.81, OA31.05 Chappell, Duane...... PS03.11, OA43.03 Cantón, Candela...... PS01.78 Charbonnel, Nathalie...... OA47.07 Cao, Jie...... OA04.05, OA17.01, OA29.03 Charlier, Johannes...... OA12.01, PS03.31 Cappiello, Briana...... OA06.06 Charnet, Pierre...... OA13.02 Caputo, Vincenzo...... PS02.15 Chartier, Christophe...... PS01.76, OA02.04, Cardim de Oliveira, Priscilla... PS03.18, PS03.82 ...... OA02.05, OA12.01, PS02.11 Cardoso, Flavia...... PS01.64 Charvet, Claude.... OA13.02, OA13.04, OA31.04, Cardoso, Luís...... OA18.02 ...... OA48.05 Cardozo, Patricia...... PS03.76 Chaudhry, Umer....PS01.77, OA24.04, OA24.05, Carey, Thomas...... OA16.04 ...... PS03.53, OA31.01, OA46.06 Carleton, Karen...... PS01.41 Chauvin, Alain...... OA02.04, OA02.05, OA12.01 Carman, Michelle...... PS02.72 Chaves, Camila...... PS01.08, OA07.06 Carnés, Jerónimo...... OA23.06 Chavéz Soriano, Larisa...... OA22.05 Carpenter, Christopher...... PS03.70 Chelladurai, Jeba J...... PS01.15 Carrera, Inés...... PS03.30 Chen, Eric...... PS03.25, PS03.35 Carrez, Florian...... PS01.65 Chen, Jing...... OA17.05 Carvalho, Itallo...... PS01.59 Chen, Jinru...... PS02.02 Casaert, Stijn...... PS03.78, OA39.06 Chen, Pai Chia Rebecca...... OA19.02 Cassaigne, Romain...... PS02.11 Chen, Rebecca...... PS03.74, OA46.01 Castagna, Fabio...... PS02.81 Chen, Ze...... OA03.05 Castilla, Verónica...... OA36.08 Chen, Zhen Zhen...... OA39.06 Castilla Gómez De Agüero, Verónica...OA41.05 Chen, Zhenzhen...... PS01.81 Castillo-Gallegos, Epigmenio.PS01.28, OA50.01 Chenafi, Ilham...... OA25.04 Castro, Pablo...... OA21.04 Chengalva Rayulu, Vukka...... OA39.07 Chevillot, Aurélie...... OA25.04, OA25.05

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 331 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Chiarotto, Francesco...... OA33.01 ...... PS02.85, PS03.43, PS03.55, PS03.60 Chigure, Gajanan...... OA49.07 Cotter, Paul...... OA38.07 Chilón, Silvia...... PS01.51 Cotton, James...... OA24.03, OA24.04, PS03.24 Chimienti, Giovanni...... OA09.04 Cotton, James...... PS01.79 Chimnoi, Wissanuwat...... PS02.36 Courtot, Elise...... OA31.04, OA48.05 Chinnaiyan, Soundararajan... PS01.52, OA40.04 Courtot, Elise...... OA13.04 Chiummo, Rafael...... PS01.30 Cox, Erin...... OA18.04, OA23.01 Cho, Yun Sang...... PS01.47, PS02.38, PS03.44 Craig, Bryant...... PS03.11, OA43.03 Choi, Ryan...... OA04.01 Cringoli, Giuseppe.OA12.01, OA20.03, PS03.07 Choocherd, Suchada...... PS02.36 Crispie, Fiona...... OA38.07 Choudhary, Shivani....PS01.15, PS01.21, OA01.06 Cristina Alves Teixeira, Marcia...... PS03.18 Christen, Isabelle...... PS03.01 Crupi, Rosalia...... PS02.81 Christina Alves Teixeira, Marcia...... PS03.82 Cruz, Breno...... PS02.22, PS02.52, PS03.64 Ciancia, Claire...... PS01.03 Cubides-Cardenas, Jaime Andres...... PS01.24, Cilek, James...... PS01.31 ...... PS02.75, PS03.79, OA50.03 Claerebout, Edwin... PS01.81, PS03.31, PS03.78, Cunningham, Mark W...... OA47.05 ...... OA39.06 Cutolo, Andre...... PS01.54, PS01.59, OA13.01, Claret, Emmanuel...... PS01.60 ...... PS02.28, PS02.82, PS03.55 Clark, Morgan...... PS03.84, OA33.03 Cutrim-Junior, José...... PS01.16 Clemons, Bambi...... OA47.05 Cwiklinski, Krystyna...... OA35.05, OA35.06 Cleveland, Christopher...... PS03.33 Cobo-González, José Manuel...... PS01.05 D Coelho Filho, Elvis...... PS03.05 Cola, D.F...... PS02.41 D. S., Malatesh...... OA42.04, PS01.86 Colella, Vito...... OA18.05, OA21.01, OA27.03, da Silva Vieira, Luiz...... PS01.70, PS02.63 ...... OA27.05 Dadhich, Abhilasha...... PS03.36 Collins, James...... OA31.02, OA32.01 Dailledouze, Aurelie...... PS02.74 Collins, Nicola...... PS01.42, PS02.64 D’Alessio, Nicola...... PS02.15 Colpitts, Julie...... OA48.07 Dalton, John...... OA35.05, OA35.06, OA45.06 Colwell, Doug...... OA41.01 Dang, Thy Anh...... OA27.03, OA27.05 Compean-Martínez, Juan E...... OA22.05 Dangoudoubiyam, Sriveny...... OA37.04 Conan, Anne...... PS01.63, PS02.58 Dantas-Torres, Filipe...... OA18.05, OA21.04 Conboy, Gary...... OA26.06, OA47.02, OA47.06 Daoust, Pierre-Yves...... OA25.07, OA47.06 Conde, Mario...... PS01.02 Daugschies, Arwid...... PS03.68 Conejeros, Ivan...... PS03.03, PS03.04 Davies, David R...... PS03.54 Conejeros, Ivan...... PS02.37 Davis, Richard...... OA22.02, OA48.06 Connelley, Timothy...... PS01.77 Davis, Travis...... OA34.05 Conrad, Patricia A...... KL01.03 Day, Katherine...... PS03.72 Constantinoiu, Con...... OA43.04 Daza, Daniel...... PS03.62 Contreras, Marinela...... OA27.02 de Almeida, Isabella Barboza...... PS03.65 Cooke, Brian...... OA17.04 de Almeida, José Eduardo Marcondes.PS01.57 Cools, Piet...... PS03.26 de Almeida Borges, Fernando...... OA12.04 Cornélie, Sylvie...... OA13.06 de Castro, Bruno...... PS01.59, PS03.55 Corner-Thomas, Rene...... OA12.02 De Marco Verissimo, Carolina...... OA35.05 Correa, Patricia...... PS01.71 de Jong, Suzanne...... OA27.01 Correia, Carolina...... OA07.02 de la Fuente, José...... OA27.02 Correia Azevedo, Thais...... PS03.82, PS03.18 de Meireles, Maria Vanuza...... OA23.03 Corripio-Miyar, Yolanda...... PS02.49, PS03.61 de Oliveira, Márcio...... OA09.03 Cortes-Rojas, Diego...... PS02.75, PS03.79, de Queiroz, Camila...... PS02.76 ...... OA50.03 de Waal, Theo...... PS02.59, OA31.03 Cortés-Vecino, Jesús A...... PS02.78 De Rijke, Jill...... OA46.01, PS03.74 Cortet, Jacques...... OA31.04 De Seram, Eranga...... PS02.76, OA36.02 Costa, Alvimar...... PS02.22, PS02.52, PS03.64 De Vidales, M...... OA26.02 Costa Junior, Livio....PS01.26, PS01.59, PS01.16,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 332 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

De Vos, Christa...... OA08.03 Doyle, Emma K...... OA35.02 De Wilde, Nathalie...... PS03.78, OA39.06 Doyle, Stephen..... PS01.79, OA24.03, OA24.04, Debièrre-Grockiego, Françoise...... OA13.02 ...... PS03.24 Decaro, Nicola...... OA18.05 Doyle, Emma...... OA07.04 Deeckens, Brenda...... PS03.55 Drake, Jason...... OA06.01, OA16.04 Defives, Chloé...... PS01.68 Dryden, Michael...... OA10.06 Del Fava, Claudia...... PS03.65 Drysdale, Orla...... OA35.05 Del Olmo, Esther...... OA36.08 Du, Aifang...... PS01.40, OA30.02 Denning, Steve...... PS01.32 Duarte, Fernanda Calvo...... PS01.57, PS02.50, Dennis, Michelle...... OA21.03 ...... PS03.65 Dentello Lustoza, Marcio...... PS03.02, OA39.05 Duarte, José...... OA09.03 Denwood, Matt...... OA02.01 Dubourdieu, Tiphaine...... OA49.06 Deplazes, P...... OA16.01 Duchateau, Luc...... PS01.81, OA39.06 Deplazes, Peter...... OA11.04, OA26.04 Dufourd, Elaudy...... OA12.01 Desquesnes, Marc...... PS02.36, PS03.58 Duggan, Vivienne...... OA38.07 Dessi, Giorgia...... PS02.15 Duhaime, Roberta...... PS02.46, PS03.59 Deterink-Damhuis, A.M.G...... OA46.05 Duhaime, Roland...... OA44.06 Devaney, Eileen...... PS01.79, OA19.03, OA24.03, Duhamel, Gerald E...... OA41.02 ...... OA24.04, PS03.24 Dumont, Caroline...... PS03.69 Diacu, Ana-Maria...... OA21.02 Dumont, Pascal...... PS02.40 Dias, Leonice...... PS02.27 Dumont, Nicolas...... PS02.74 Dias, Lucas Pinheiro...... PS01.25 Duncan, Kathryn...... OA03.01, OA03.02 Dias de Castro, Luciana...... OA38.08 Duniho, Steve...... OA14.04 Díaz, André...... PS01.75 Duscher, Georg...... OA27.02 Diedrich, Jolene...... OA08.01, OA08.02 Duvallet, Gerad...... PS03.58 Diekmann, Irina...... OA26.02 Dyachenko, Viktor...... PS03.68 DiGennaro, Peter...... OA06.04 Dzimianski, Michael...... PS01.07 Dilks, Clayton...... OA19.01 Dillio, João...... PS01.46 E Dimalibot, Judeline...... PS02.88 Dimier-Poisson, Isabelle...... OA13.02 Eastwood, Steve...... OA14.03 Dini, Pouya...... OA48.06 Ebbesen, H. H...... OA16.01 Diuk-Wasser, Maria...... OA44.04 Echaides Ortiz, Cèsar Fernando...... PS02.32 Divis, Paul Cliff Simon...... OA40.01 Echaides Ortíz, César...... PS02.31 Dixon, Brent...... OA25.01 Edgley, Mark...... PS01.72 Długosz, Ewa...... PS02.66 Edna Tomiko Myiake, Kato...... PS03.62 Dobson, Robert...... OA02.01 Edquiban, Tisha Rogelle...... OA39.02 Dolon, Magali...... PS01.01 Eduardo Ramirez Espana, Eduardo.....OA39.03 Domingos Ferrari, Elis...... PS01.38 Ehtisham-ul-Haque, Syed...... OA21.07, OA50.07 Domingues, Luísa N...... OA34.01, OA34.02 Eichenberger, Ramon Marc...... OA22.01 Dominguez, Paula...... PS01.82 Ekanayake, Samantha...... PS02.76, OA36.02 Domínguez, Laura...... PS03.30 Elfawal, Mostafa...... OA22.03, OA28.02 Domínguez, Maria Paula...... OA39.08, PS03.81, Elkin, Brett...... PS03.32 ...... OA31.05 Ellison, Siobhan...... OA14.01, OA14.04 Domínguez, Paula...... PS01.78, PS02.83 Elmahalawy, Safaa T...... OA07.07 Donahue, Bill...... PS01.06 Elochukwu, Victoria...... PS01.56 Donaldson, Taylor...... PS01.44 Elsemore, David.. OA05.03, OA10.03, OA10.04, Dong, Junyan...... OA27.03, OA27.05 ...... OA10.05, PS02.51, OA16.03 Donnelly, Sheila...... OA35.05 Elsheikha, Hany...... OA35.06, OA45.05 Dorr, Carlie S...... OA04.01 Elsheikha, Hany M...... OA04.06, OA27.04 dos Anjos, Karina Araújo...... PS01.57, PS02.50, El-Shemy, Ahmed...... OA37.01 ...... PS03.65 Emery, David...... OA35.04 Douhard, Frederic...... PS01.09 Engborg, Steven...... OA44.06 Douine, Camille...... PS03.69

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 333 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Enzelberger, Manuela...... PS02.51 Filia, Gursimran...... PS01.58 Epe, Christian...... OA44.01 Fioretti, Alessandro...... PS02.15 Epperson, William...... OA36.03 Fiorin, Cristiane Fernandes de Carvalho...... Escareño-Díaz, Selene...... OA50.01 ...... PS01.74, PS02.23, PS02.73, PS03.51 Esteve–Gasent, Maria D...... PS02.62 Fiorini, Leonardo Costa...... PS01.57, PS02.50, Estrada, Marko M...... OA42.03 ...... PS03.65 Etienne, Lucas...... PS03.58 Fischer, Katja...... OA13.05 Etter, Eric...... OA25.06 Fisher, Maggie...... PS01.62 Eugin D’Souza, Placid...... OA40.07 Flanagan, Kelly...... OA28.02, OA50.02 Evangelista, Jayvee...... OA39.02 Florian, Untersweg...... PS02.77 Evans, Alec...... OA11.03 Flórid, João Gabriel...... PS03.64 Evans, Christopher...... PS01.07, PS03.72 Flota Burgos, Gabriela Janett...... PS01.27 Evans, Mike...... PS01.77, OA31.01, OA46.06 Foglia Manzillo, Valentina....OA23.02, OA23.05 Eydal, Matthías...... OA48.03, OA48.04 Foil, Lane D...... OA34.01 Ezeokonkwo, Romanus...... PS02.86 Foj, Rubén...... OA23.06 Follet, Jérôme...... OA25.04, OA25.05 F Foltz, Jennifer...... OA18.04 Fonseca, Dina...... OA03.04 F. Andersen, John...... PS01.13 Fonseca, Evelise...... PS02.83 F. Urban, Joseph...... OA28.02 Fontes, Florencia...... PS01.75 Faciola, Antonio...... PS01.71 Forzán, María J...... OA47.04 Failing, Klaus...... PS01.67 Foulk, Donna...... OA15.01 Fairman, Donald...... PS02.69 Fourie, Josephus...... OA11.03, OA39.04 Fairweather, Ian...... OA35.01 Fournel, Sandrine...... PS01.34 Falcão, Alexandre Xavier...... OA37.03 Fowler, Fallon...... PS01.32 Fang, Fang...... PS02.10, OA27.03, OA27.05 Fowler, Hailie...... PS01.69 Fankhauser, Becky.PS02.03, PS03.83, OA36.05 Fowler, Justin...... OA32.01 Faria Silva, Marco Antonio...... PS03.51 Fraceto, L.F...... PS02.41 Farias, Cristina...... PS01.82, PS03.76 Franc, Michel...... OA13.06, PS03.58 Farougou Souaibou...... PS03.47 Freitas, Mariana...... PS01.02, PS03.05 Farries, Gabriella...... OA07.02 Freitas, Filipe...... PS03.55 Fatoba, Abiodun...... PS02.35 Frey, Caroline...... OA20.01, OA40.06 Faulkner, Charles...... OA06.06 Friesen, Kristina...... PS02.33 Fávero, Flávia...... PS03.05 Fryxell, Rebecca...... OA34.05 Feitosa, Roseane...... OA23.03 Fujisaki, Kozo...... OA03.06 Felippelli, Gustavo.PS02.22, PS02.52, PS03.64 Furlanello, Tommaso...... OA18.05 Feng, Yaoyu...... OA25.02 Fuwa, Yui...... OA40.05 Fernandes, Julio...... PS03.80 Fernandes, Murilo Antonio...... PS03.56 G Fernandez, Christina...... OA06.08 Fernandez, Maria del Pilar...... OA44.04 G. B., Manjunath Reddy...... OA42.04, PS01.86 Fernandez, Mônica...... PS01.20 Gabor, Leslie...... OA35.02 Fernandez, Sandra...... PS02.83 Gadanho, Marion...... OA12.01 Fernandez, Silvina....PS01.83, PS02.47, PS03.57 Gainza, Y.A...... PS02.41 Ferreira de Oliveira, Gabriela...... PS03.02, Galan, Maxime...... OA47.07 ...... OA39.05 Galay, Remil...... OA03.06, OA39.02 Ferreira Gomes, Jancarlo...... PS01.38 Gall, Cory...... PS01.42 Ferreira Vasconcelos, Janaelia...... PS01.70, Gallina, Tiago...... OA39.01 ...... PS02.24 Gallo, Kendra...... PS03.25 Ferrer, Lluis...... OA23.06 Gamalo, Lief Erikson...... PS02.88 Ferrer, Manuela...... PS03.30 Gamboa Angulo, María Marcela...... PS01.27 Fiel, Cesar...... OA39.08 Gao, Wenjie...... OA24.08 Figueiredo, Amanda...... PS02.41 Garcia, Marcos...... PS01.46

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 334 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Garcia-Campos, Andres...... OA07.02, PS02.59 Goni Dogo, Abraham...... OA15.04 García-Hernández, Ángel...... PS01.05 González, Jorge F...... PS02.49, OA07.01, Garner, Bridget...... PS03.72 ...... PS03.61, OA41.05 Garza-Cuartero, Laura...... OA07.02, OA45.01, Good, Barbara...... OA31.03 ...... OA45.06 Good, Liam...... PS03.85 Gasbarre, Louis...... OA36.03 Goodman, Laura B...... OA43.01 Gasibauer, Stefanie...... PS03.01 Goodrich, Erin L...... OA43.01 Gasser, Robin B...... PS01.45, OA19.06 Goolsby, John...... PS03.59 Gaughan, Earl...... PS03.11, OA43.03 Gozalez-Pech, Pedro...... PS01.16 Gavriliuc, Stefan...... PS03.74 Graff, Emily...... PS01.39 Gazzola, David...... OA28.02, OA50.02 Graham, David...... OA45.01 Geary, Timothy...... PL01.01, OA01.04, OA46.03 Grandi, Giulio...... PS02.04 Geden, Christopher...... OA32.04 Grant, William...... PS01.44, OA49.02 Geib, Scott...... PS02.30 Gravatte, Holli...... OA33.03, OA48.06 Gelasakis, Athanasios...... PS01.80, PS03.63 Gray, Dawn...... OA41.01 Geldhof, Peter...... PS03.78 Gray, Jeremy...... OA49.06 Geng, Jinming...... OA10.05, PS02.51 Greenberg, Hannah...... OA09.02 Genro, Jonis...... PS03.05 Greenwood, Spencer...... OA26.06, OA47.02, George, Ashley...... OA35.04 ...... OA47.06 George, Sarah...... OA35.04 Greer, Andrew...... OA02.02, PS03.15 Geraldes Teixeira, Marta M...... OA29.01 Gregory, Lilian...... PS03.62 Gerhard, Alexander...... OA22.02 Griffith, A. .P...... OA34.04 Gerhold, Richard...... OA30.01 Grinnage-Pulley, Tara...... PS01.69 Gerry, Alec...... OA32.03 Grisez, Christelle.....PS01.09, PS02.74, PS03.58, Gettings, Jenna...... OA01.01, PS02.70, PS03.70 ...... OA46.02 Geurden, Thomas...... OA45.06 Gruntmeir, Jeff...... OA06.05 Geyer, Kathy...... OA33.06 Gruszynski, Karen...... OA06.06 Ghafar, Abdul...... PS01.45 Guechi, Radia...... PS01.68 Gharbi, Mohamed...... PS01.33 Guégnard, Fabrice...... OA31.04 Ghosh, Parna...... OA03.01, OA03.03 Gueijman, Jordana...... PS02.53 Ghosh, Srikant...... OA08.04, OA49.07 Gueldner, Emily...... PS03.67, OA06.07 Gibson-Corley, Katherine...... OA23.01 Guerin, Michele...... PS03.73 Giglioti, Rodrigo...... PS01.22, PS01.48, PS01.74, Guerra, Nicolas...... OA04.04 PS02.23, PS02.44, PS02.45 PS02.73, PS03.28, Guerrero, Felix...... PS02.30, OA34.01, OA49.04 ...... PS03.49, PS03.50, PS03.51 Guerrero, Inés...... PS02.47, PS03.57 Gilleard, John...... PL02.04, PS01.72, OA24.04, Guerrero-Molina, Cristina...... PS01.05, PS01.11, ...... PS03.24, PS03.26, PS03.74, ...... PS03.14 ...... OA19.02, OA24.05, OA36.02, Guillot, Jacques...... PS01.68, OA13.05, PS02.10, ...... OA38.05, OA46.01, OA48.07, PS01.17, ...... OA42.02 ...... PS02.76, OA24.06, OA26.05, OA41.01 Guimaraes, Paulo...... OA38.08 Gilsdorf, Cole...... PS03.25 Guo, Aijiang...... PS02.17 Giroto, Laura...... PS03.64 Guo, Fengguang...... OA46.04 Giuffrida, Rogerio...... PS02.27 Guo, Junhui...... OA03.05 Gizzarelli, Manuela...... OA23.02, OA23.05 Guo, Xiaola...... PS03.17 Goddu, Troy...... OA10.02 Gustafsson, Katarina...... OA07.07 Gokool, Suzanne...... PS02.16 Gustinelli, Andrea...... OA09.04 Golfinos, Athena...... PS03.25 Gutierrez Gil, Beatriz...... OA41.05 Gomes, Jancarlo Ferreira...... OA37.03 Gutmanis, Gunta...... PS02.23, PS02.73, PS03.51 Gomes, Lucas...... PS02.22, PS02.52, PS03.64 Gomes, Matheus Nobate...... PS03.60 H Gómez, Perla...... PS02.80 Gomez-Alvarez, Martha...... PS02.75, PS03.79 H. Pihl, Tina...... PS02.08 Gómez-Alvarez, Martha Izabel...... OA50.03 Habing, Greg...... PS02.72 Gong, Haiyan...... OA29.03

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 335 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Hadfield, Kerry .A...... PS03.37, OA37.02 Herzig, Volker...... OA50.06 Hafeez, Rehman...... OA49.05 Hess, Lindie...... OA16.02 Hairgrove, Thomas...... OA49.01 Hessle, Anna...... OA02.03 Halos, Lénaïg...... OA27.03, OA27.05 Heuer, Lea...... OA40.02 Halvarsson, Peter.PS02.07, OA07.07, OA24.02, Hickling, Graham...... OA09.01, OA30.01 ...... OA36.06 Higa, Leandro...... PS01.46 Hamel, Dietmar...... PS02.03, OA36.05 Hilary, Mailys...... PS02.34 Hamer, Kim...... PS01.79 Hilbe, Monika...... OA06.07 Hamer, Sarah...... PL04.04 Hildreth, Michael B...... PS02.12 Hamza, Talel...... OA27.06 Hine, Brad...... OA07.04 Han, Qian...... OA17.05 Hinkle, Nancy C...... PS02.02 Hanna, Rita...... OA16.03 Hinney, Barbara...... OA04.04, PS02.77, PS03.01 Hanscom, Jancy...... OA05.03, OA27.04 Hirschauer, Léa...... PS01.76 Hardham, John M...... OA45.06 Hobán, Cristian...... PS01.50, PS01.51 Hargis, Billy...... PS03.73 Hoberg, Eric...... PS03.32 Harms, N Jane...... PS03.32 Hockley, Tamara...... OA05.03, OA06.08 Harrison, Mark...... PS02.02 Hodgkinson, Jane...... , OA24.07, PS03.54 Hasel, Hallie...... PS03.59, OA49.02 Hodo, Carolyn...... OA47.05 Hastriter, Michael...... PS02.18 Hodžić, Adnan...... OA04.04, OA27.02 Hatta, Takeshi...... OA03.06 Hofmar-Glennon, Hayley G...... OA43.01 Hayat, Khizer...... OA17.06 Högberg, Niclas...... OA02.03, PS02.04 Hays, Samantha...... OA49.01 Höglund, Johan....OA02.03, OA07.07, PS02.04, He, Jun-Jun...... OA04.06, OA45.04, OA45.05 ...... PS02.07, OA24.02, OA22.08, Heaney, Kathy...... OA10.07 ...... OA36.06, OA48.03, OA48.04 Hebert, Deborah...... OA12.05 Hogsette, Jerome...... PS01.31 Hebron, Claude...... OA32.01 Hoisington, Aaron...... OA01.06 Heckeroth, Anja...... PS01.30 Holderman, Chris...... OA06.04 Hedberg-Alm, Ylva...... OA33.05 Holland, Celia...... OA47.07 Heesen, Henk...... OA27.01 Holroyd, Nancy..... OA24.03, OA24.04, PS01.79, Hegarty, Evan...... OA10.06 ...... PS03.24 Hegglin, Daniel...... OA26.04 Homer, David...... OA02.06 Heidmann, Maycon...... PS01.59, PS03.55 Honoré Jørgensen, Marie Louise...... PS02.08 Heimark, Zachary...... PS01.18, PS03.35 Hoste, Herve...... PS03.31 Heitlinger, Emanuel...... OA22.02 Howe, Daniel...... OA37.04, OA48.06 Heliso, Priscilla S...... PS02.12 Howell, Rebecca...... PS01.66 Helm, Christina...... OA11.04 Howell, Sue...... OA31.02 Helps, Chris...... OA44.05 Hoyle, Rebecca...... OA24.07 Hemphill, Andrew...... OA04.01 Hu, Min...... OA24.08 Hendawy, Seham...... OA42.01 Hu, Rui-Si...... OA35.06, OA45.05 Henriques, Paula...... PS01.08, OA07.06 Hu, Yan...... OA28.02, OA50.02 Henttonen, Heikki...... OA47.07 Huang, Si-yang...... OA45.02 Hermosilla, Carlos.....PS01.37, PS01.67, PS02.37, Huang, Yu-Hsin...... OA19.01 ...... PS03.03, PS03.04 Hubert, Floriane...... PS01.65 Hernandez, Emmanuel...... OA03.06 Hudson, Ariel...... OA06.06 Hernandez, Leticia...... OA26.02 Huelle, A. R...... OA16.01 Hernández, Julia Natividad... OA07.0, OA41.05, Hughes, Kris...... OA15.02 ...... PS02.49, PS03.61 Hulverson, Matthew A...... OA04.01 Hernandez Ortiz, Adrian...... OA25.07 Hunter, Alexa...... OA11.01 Hernández-Aranda, Itzel...... PS01.05 Hurtig, Frank...... PS03.08 Herrero, Silvia...... OA26.02 Hussain, Kashif...... Herrick, Katherine...... OA21.05 Hyun, Bang-Hun...... PS01.47, PS02.38, PS03.44 Herrin, Brian...... PS01.61, PS02.01, OA10.06, ...... PS03.11, OA43.03 Hervet , Caroline...... OA13.04

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 336 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

I Jenkins, Emily...... OA20.01, OA25.07, PS03.32, ...... OA40.06, OA48.07 Ianniello, Davide...... OA20.03, PS03.07 Jenkins, Timothy...... OA43.06 Iatta, Roberta...... OA18.05 Jesudoss Chelladurai, Jeba R J...... OA16.05, Ichikawa-Seki, Madoka...... PS01.50 ...... PS03.05, PS03.27 Idika, Kalu...... PS02.86 Jewhurst, Heather...... OA35.05 Ientile, Michelle...... OA03.01, OA03.03, PS02.42 Jimenez Castro, Pablo David...... OA26.05 Iglesias, Lucía...... PS01.83, PS02.47, PS03.57 Jin, Zi...... OA46.04 Ijaz, Farah...... , OA21.07 Joachim, Anja.... OA04.01, OA04.03, OA04.04, IJzermans, Carlijn...... OA27.01 ...... PS03.01, PS02.77 Ikurior, Seer...... OA12.02 Joanes, Sloane...... PS01.34, PS03.08 Iliev, Petar...... PS03.83 John, Bethan...... PS03.54 Imai, Rachel...... OA37.05, OA37.06 Johne, Annette...... OA30.04 Imato, Reinaldo...... PS02.83 Johnson, Eileen...... OA21.05 Imparato, Laetitia...... PS01.65, PS02.34 Jones, Brittany...... PS03.84 Imperiale, Fernanda...... PS01.82 Jongejan, Frans...... OA27.01, OA39.04 Inácio, Sandra Valéria...... OA37.03 Jonk, Kaitlyn...... PS01.63 Inglese, Antonio...... OA23.05 Jonsson, Sofia...... OA48.02, OA48.03 Innocent, Giles...... OA19.02 Jordan, Brian...... OA32.01 Inpankaew, Tawin... PS01.84, PS02.36, OA21.04 Jordy, Julia...... OA03.01 Ionică, Angela Monica...... OA21.02 Joshua, Kamani...... OA21.04 Iqbal, Khalid...... OA50.05 Julien, Christine...... OA25.04 Iqbal, Zafar...... OA17.06, OA28.04, OA32.05, Juliet, Sanis...... OA49.07 ...... OA49.05 Junco, Milagros...... PS01.83 Irwin, Peter...... OA21.04 Jürgenschellert, Laura...... PS03.13 Isaksson, Mats...... OA20.02 Isloor, Shrikrishna...... OA40.07 K Issakowicz, Juliano...... PS01.71 Issouf, Mohamed...... PS01.19 K, Sumana...... PS01.86, OA42.04 Ivanek, Renata...... OA43.01 K Javaregowda, Ananda...... OA42.04 K. Nielsen, Martin...... PS02.08, OA28.02 J K. V., Lavanya...... OA42.04, PS01.86 Kadir, Khamisah Abdul...... OA40.01, PS02.88 Jabbar, Abdul...... PS01.45, OA14.02, OA41.04 Kahn, Lewis...... OA07.04 Jäckel, Claudia...... OA30.04 Kalayou, Shewit...... OA12.06, PS02.60 Jackson, Andrew...... OA22.06, OA29.01 Kalin, Haluk...... PS02.19, PS03.20 Jackson, Joeseph...... OA47.07 Kamyingkird, Ketsarin...... PS02.36 Jacobsen, Stine...... PS02.08 Kandasamy Perumal, Prabhu...... OA40.04 Jacquiet, Philippe...PS01.09, PS02.74, PS03.58, Kandil, Omnia...... OA42.01 ...... OA46.02 Kaplan, Ray...... OA02.01, OA12.03, OA26.05, Jähne, Stéphanie...... PS03.01 ...... OA31.02, OA32.01 Jaleta, Tegegn...... PS02.21 Karam, Gina...... OA34.06 James, Peter...... OA43.04 Karembe, Hamadi...... OA04.04 Janecek-Erfurth, Elisabeth...... OA40.02 Karlsson Lindsjö, Oskar...... OA48.03, OA48.04 Jantanawaranon, Thunchanok...... PS01.49 Karrow, Niel...... PS02.20, PS03.22 Jardine, Claire...... OA20.02 Karteri, Olga...... PS03.75 Jarriel, Dixie...... OA16.03 Kashyap, Sudhanva.PS01.21, OA01.06, OA19.05, Jedrzejewski, Ed...... OA15.01 ...... OA19.04, OA22.04 Jegede, Cornelius Olorunfemi...... , PS01.56 Kastelein , Lisette...... OA46.05 Jegede, Olorunfemi.PS01.55, PS02.48, PS03.12, Kästner, Carolyn...... OA30.04 ...... OA29.02 Katiki, Luciana...... PS01.08, PS01.22, PS01.74, Jemli, Mohamed Habib...... OA27.06 OA07.06, PS01.48, PS02.45, PS02.23, PS02.27, Jenkins, Cheryl...... OA17.03 PS02.73, PS02.84, PS03.28, PS03.50, PS03.51,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 337 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

...... PS03.62, PS03.56 ...... PS01.86, OA08.04, OA40.07 Kaufman, Phillip...... OA06.04 Konecsni, Kelly...... PS03.32 Kaur, Paramjit...... PS01.58 Kontowicz, Eric...... PS01.69 Kavitha Rani, B...... PS01.12 Kopsco, Heather...... OA44.06 Keane, Orla...... OA31.03 Korhonen, Pasi...... OA19.06 Kearns, Kelsey...... PS01.17 Koser, Troy...... PS03.33 Kebeta, Mulisa...... OA07.04 Koston, Florian...... OA04.01 Kelleher, Anne...... OA31.03 Kotwa, Jonathon...... OA20.02 Keller, Kathy...... OA01.04 Kotze, Andrew C...... OA50.06 Kellermann, Michael...... OA36.05 Koziar, Nadia...... OA13.03 Kellogg, Tasia...... OA28.02, OA50.02 Krecek, Rosina...... KL01.03, PS01.62, PS02.58 Kelly, Aubree...... PS02.02 Krishnavajhala, Aparna...... PS02.70 Kelly, Gareth...... OA02.06 Krücken, Jürgen.... OA11.04, OA12.04, OA22.02, Kelly, Janetta...... OA30.01 ...... OA26.02, PS03.13 Kemble, Nick...... PS03.21 Krueger, Christian...... PS03.23 Kenealy, Jessica...... OA33.03, PS02.25 Kruth, Perryn...... PS01.53, PS02.67, PS03.73, Kenney, Daniel...... PS01.41 ...... OA37.05 Kent, Michael...... OA15.03 Kryda, Kristina...... OA01.02 Keogh, Eammon...... OA32.03 Kudrna, Katherine...... PS01.18 Kerner, Katharina...... PS03.03 Kulasooriya, Bhagya...... OA08.07 Kessa, Haykel...... OA27.06 Kulich, Pavel...... PS02.43 Kesumawati, Upik...... OA27.03, OA27.05 Kumar, Binod...... OA49.07 Ketzis, Jennifer...... PS01.63, OA10.05, PS02.09, Kumar, Rajesh...... OA49.07 ...... OA21.03, OA21.04, OA28.02 Kumar, Satyanshu...... OA49.07 Kgomotso, Sibeko P...... OA29.04 Kumaresan, Nagarajan...... PS01.52, OA40.04 Khan, Adil...... PS01.45 Kunkle, Bruce...... PS02.03 Khan, Muhammad Azmat Ullah...... PS01.77 Künzel, Frank...... PS03.01 Khan, Muhammad Kasib...... OA49.05 Kurzrock, Lina...... OA26.02 Khanal, Pratisksha...... PS02.12 Kusakisako, Kodai...... OA03.06 Khatiworavage, Chatree...... PS01.49 Kuzmina, Tetiana...... OA12.04 Kheravii, Sarbast...... PS03.39 Kuzyk, Amanda...... PS01.17 Kidd, Hayley...... OA43.04 Kvaternick, Valerie...... OA36.05 Kim, Chi Yong...... OA04.02 Kwon Kim, Tae...... OA08.01 Kim, Tae Kwon...... OA08.02 Kimber, Michael...... PL02.02 L King, Glenn F...... OA50.06 King, Vickie...... PS02.01 Labarthe, Norma....PS01.20, PS01.64, PS03.80, Kirkova, Zvezdelina...... PS03.83 ...... OA23.04 Kirkvold, Emily...... PS02.33 Labuschagne, Michel...... OA11.03 Kitchen, Shannon...... PS02.21 Lahouar, Ahmed...... OA27.06 Klafke, Guilherme.OA03.04, PS02.78, PS03.59, Laing, Roz...... PS01.79, OA19.03, OA24.03, ...... OA39.01, PS01.75, PS03.77 ...... OA24.04, PS03.24 Kline, Daniel...... OA43.05 Lakew, Biniam...... OA14.03, PS02.29, PS03.39 Klockiewicz, Maciej...... PS02.66 Lakritz, Jeffrey...... PS02.72 Kloc-Muniak, Klaudia...... PS03.85 Lamassiaude, Nicolas...... OA13.02, OA48.05 Klubkova, Valeriia...... PS01.03 Lamb, Jennifer...... OA45.03 Knaus, Martin...... PS03.29 Landulfo, Gabriel Alves...... PS03.43 Knupp, Andrew...... OA08.07 Lansford, T.R...... PS03.59 Kockmann, Tobias ...... PS03.09 Lanusse, Carlos...... PS01.78, PS01.82, PS02.83, Koehler, Anson...... OA19.06 PS02.85, PS03.76, PS03.81, OA31.05, OA35.01, Kohn, Barbara...... OA11.04 ...... OA39.08 Kolapo, Temitope...... OA20.01, OA40.06 Lanza Perea, Marta...... PS01.66, OA27.04 Kolo, Agatha...... PS01.42 Larson, Mandy...... PS01.69 Konanur Javaregowda, Ananda...... PS01.12,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 338 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Larter, N C...... PS03.32 ...... PS02.85, PS03.76, OA39.08 Latrofa, Maria Stefania...... OA09.04, OA18.05 Lightbody, Kirsty...... OA33.06 Lavan, Robert...... OA01.03, OA01.05, OA10.07 Lightowlers, Marshall...... OA20.04, OA40.03 Lawal, Idris...... OA29.02 Lillie, Brandon...... PS02.20, PS03.22 Lawlor, Amanda...... PS02.59 Lim, Jiyeon...... PS01.47, PS03.44, PS02.38 Lawton, Colin...... OA47.07 Lima, Adam...... PS01.69 Le, Thong Quang...... OA27.03, OA27.05 Lima, Aldilene Silva...... PS03.43 Leathwick, Dave...... PS03.34, OA07.03, Lima, Ruana Cardoso...... PS01.25 ...... OA09.05, OA38.01, OA38.02, Lin, Dan...... OA33.02 ...... OA38.03, OA38.04, OA38.06, OA41.03 Lin, Xiaoxing...... OA04.05 Lebepe, Jeffrey...... PS02.35 Linh, Bui...... OA21.04 Lebon, Wilfried...... PS01.43, OA08.03, PS02.40 Lins, Talita...... OA23.03 Lee, Dean...... OA18.04 Liotta, Janice L...... OA21.06 Lee, Hyunkyoung...... PS01.47, PS03.44 Lipworth, Karen...... OA01.03 Lee, Xia...... OA11.05 Lirón, Pedro...... PS01.78 Lee, Yuri...... OA21.06 Little, Susan...... OA03.01, OA03.02, OA03.03, Lee, Seung-Hun...... PS01.35 ...... OA08.07, OA10.02, OA11.01, PS02.42, Lees, Jeff...... PS01.17 ...... OA26.03, PS03.10 Lehebel, Anne...... OA02.04, OA02.05, OA12.01 Liu, Chunliang...... OA36.07 Leite, Paula...... PS02.83 Liu, Guangxue...... PS03.17 Lejeune, Manigandan...... PS02.61, OA41.02, Liu, Guangyuan...... OA03.05 ...... OA43.01, OA47.04 Liu, Mingming...... PS01.36 Lemler, Erica...... OA05.04 Liu, Tony Chun Yu...... OA36.07 Lemos, Leandro...... PS01.71 Liu, Wenge...... OA03.05 Lempereur, Laetitia...... PS03.47 Liu, Xin...... OA24.08, OA27.03, OA27.05 Leschnik, Michael...... OA27.02 Lizundia, Regina...... OA44.01 Lespine, Anne...... PS02.74 Lloberas, Mercedes...... PS02.85, PS03.76 Lessey, Lindyann...... OA06.08 Loghry, Hannah...... PL02.02 Leu, Stephan...... OA12.02 Login, Joyce...... OA13.03 Leung, Tommy L.F...... OA35.02 Lohmeyer, Kimberly...... OA49.02 Leutenegger, Christian...... PS01.66, OA05.03, Loiola, Saulo Hudson Nery...... OA37.03 ...... OA05.05, OA26.01, OA27.04, OA42.03 Loiseaux, Juliette...... PS01.68 Levecke, Bruno...... OA02.01, PS03.26, PS03.78 Lok, James...... PS02.21 Léveillé, Alexandre...... PS01.41, PS03.41 Long, Maureen...... OA06.05 Levy, Michel...... PS01.17, OA24.06 Long, Sidney...... PS02.57 Lewis, K...... OA34.04 Lopes, Welber...... PS02.22, PS02.52, PS03.64, Lewis, Nicole...... PL04.02 ...... OA09.03 Li, Hanchen...... OA28.02 Lopez, Javier...... OA26.02 Li, Kun...... OA04.02, OA09.07 Lopez, Job...... PS02.70 Li, Ting-Ting...... OA04.06 Lopez Osorio, Sara... PS01.37, PS01.67, PS02.37 Li, Hanchen...... OA50.02 Lopez-Muñoz, Rodrigo...... OA28.03 Lia, Riccardo P...... PS03.38 Lorusso, Eleonora...... OA18.05 Liang, Panhong...... PS03.17 Lorusso, Vincenzo...... OA15.04 Liang, Qin-Li...... OA04.06 Loseto, Lisa...... OA25.07 Liao, Chenghong...... OA17.05 Loukas, Alex...... OA22.01 Liboiron, Alice...... OA48.07 Loukeri, Sofia...... PS01.60, PS03.63 Lidfors, Lena...... OA02.03 Louro, Mariana...... OA12.04 Liebenberg, Julian...... PS01.43 Louvandini, Helder....PS01.08, PS01.16, PS01.22, Liebstein, Martin...... PS02.03 PS01.71, OA07.06, PS02.14, PS02.23, PS03.28, . Lien, Yi-Yang...... OA08.05 ...... PS03.56 ...... Loxton, Karen...... OA47.07 Liénard, Emmanuel ...... OA13.06, PS03.58 Lozano Blanco, Ivan...... OA39.03 Liesner, Jana...... OA11.04 Lu, Jinmiao...... OA29.03 Lifschitz, Adrián...... PS01.78, PS01.83, PS02.83, Lu, Na...... OA27.03, OA27.05

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 339 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Lubembe, Donald Mukolwe...... OA29.04 Maloney, Jenny...... OA25.03, OA42.06 Luciani, Guilherme Favero...... PS01.22, PS01.74, Mamman, Adamu Haruna...... OA15.04 ...... PS02.23, PS02.73, PS03.28, PS03.51 Mammeri, Mohamed...... OA25.04, OA25.05 Lucio-Forster, Araceli...... OA47.03 Mangelsdorf, David...... PS02.21 Lühder, Fred...... OA40.02 Mani, Arul Prakash...... PS01.52, OA40.04 Lund, Robert...... PS03.70 Manjunath, S. S...... PS01.12 Lunn, Nicholas...... OA25.07 Manojai, Nuttaphon...... PS02.36 Luo, Jianxun...... OA03.05 Mao, Li...... PS03.17 Luo, Jin...... OA03.05 Maqbool, Mahvish...... PS03.48, OA50.03 Luo, Xuenong...... PS02.17, PS03.17 Marcon, Didier...... PS01.09, OA46.02 Luo, Houqiang...... OA09.07 Maree, Riaan...... OA11.03 Lúpori, Jorgelina...... PS01.82 Marendy, Dominique Maree...... OA35.02 Lupu, Andrei Cristian...... OA13.05 Maria Leal Bevilaqua, Cláudia...... PS01.70 Luque, Sonia...... PS02.85, PS03.76 Maria Leite dos Santos, Jessica...... PS01.70, Lustoza, Marcio...... PS03.18 ...... PS02.63 Lyons, Eugene...... PS03.84 Markl, Andrea...... PS03.01 Lyubomska, Oksana...... OA35.05 Marquetoux, Nelly...... OA12.02 Marsh, Antoinette...OA15.05, PS02.57, PS02.72, M ...... OA16.02 Martin, Donald...... OA10.02 M Kukka, Piia...... PS03.32 Martin, Frida...... OA48.02, OA48.03, OA48.04 M. C. Ribeiro, Jose...... PS01.13 Martin, Katherine...... PS03.27 Ma, Guangxu...... OA19.06 Martin, Katy...... PS03.05 Ma, Jun...... OA45.04 Martin, Richard...... PS01.15, PS01.21, OA01.06, Mabelebele, Monnye...... PS03.16 ...... OA19.04, OA19.05, OA22.04 Macedo, Eduardo Pires...... PS03.65 Martínez, Carlos...... PS01.24 Macedo de Toledo, Luciandra...... PS02.73 Martínez, María...... OA36.08 Machin, Cynthia...... PS02.49, PS03.61 Martínez Ortiz de Montellano, Cintli...... PS01.11, Machtinger, Erika.OA09.01, OA09.02, OA32.04 ...... PS01.27 MacHugh, David...... OA07.02 Martinez Valladares, Maria.....OA35.01, OA41.05 Maciel, Willian...... PS01.46, PS02.22, PS02.52, Martínez-M, Imelda...... PS01.73 ...... PS03.64 Martinez-Orellana, Pamela....PS02.65, OA18.03, MacLeay, Madison...... OA31.01 ...... PS03.85 Madder, Maxime...... OA11.03, OA39.04 Martins, Isabela...... PS02.82 Madeira de Carvalho, Luís M...... OA12.04 Masiga, Dan...... OA12.06 Madikizela, Balungile...... PS03.16 Masonick, Paul...... OA34.05 Madouasse, Aurélien...... OA02.04, OA02.05 Masuda, Aya...... OA40.05 Magalhães, Caroline...... PS01.64 Maté, Laura...... PS01.78 Magiera, Krystal...... PS03.27 Mateos-Hernández, Lourdes...... OA27.02 Maguire, Danielle...... PS02.61 Mather, Paula-Marie...... OA47.02 Mahabir, Sean...... OA01.02 Mather, Thomas...... OA44.06 Mahachi, Kurayi...... PS01.69, OA18.04, OA23.01 Mathur, Manisha...... PS03.36 Mahajan, Vishal...... PS01.58 Matjila, Paul...... OA11.02 Mahan, Suman M...... OA45.06 Matsumoto, Jun...... OA40.05 Mahjoub, Haifaa...... OA47.02 Matthes, Hans-Frieder...... OA11.04 Mahmoud, Bassma...... OA42.01 Matthews, Jacqueline B...... PS02.49, PS03.61 Mahmoud, Mona...... OA42.01 Matthews, Jacqui...... OA33.06 Mains, Paul...... PS01.72 Mattos, Waldssimiler Teixeira de...... PS01.74 Mair, Gunnar...... OA01.06 Matusop, Ahmad...... OA40.01 Maitland, Kirsty.... PS01.79, OA24.03, OA24.04, Mauldin Pereira, Mary...... PS01.63, PS02.58 ...... PS03.24 Maurelli, Maria Paola...... OA12.01, OA20.03, Malatji, Pulane...... OA45.03 ...... PS03.07 Malik, Muhammad Abdullah...... OA17.06 Maurer, Juliana...... OA50.04 Mayer-Scholl, Anne...... OA30.04

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 340 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Mayr, Sandra...... OA36.05 Neupane, Saraswoti...... PS02.71 Mazaleski, Mark...... OA01.02 Neves, Luis...... OA25.06 Mazeri, Stella...... PS03.53 Neveu, Cédric...... PS01.19, OA13.02, OA13.04, Mazzeo, Giuseppe...... OA33.01 ...... OA31.04, OA48.05 McAloon, Conor...... OA45.01 Newbury, Hannah...... OA01.03, OA44.05 McAnulty, Robin...... OA02.02, PS03.15 Newcomb, Harold...... OA36.03 McCarthy, G...... PS02.59 Newton, Jason...... OA49.06 McCrann, DJ...... OA10.02 Ngasaman, Ruttayaporn...... PS02.36 McGaw, Lyndy...... PS02.13 Nguyen, Viet Linh...... OA27.03, OA27.05 McGeachy, David...... OA25.07 Ni, Jun...... OA03.05 McGee, Evan...... PS02.69 Nicholas, Adrian H...... PS02.29 McGhee, James...... PS01.72 Nicolas, Céline...... PS01.34 McGoldrick, James...... PS01.79 Nie, Shuai...... OA19.06 McGowan, Michael...... OA43.04 Nielsen, Martin...... OA02.01, OA15.01, PS02.25, McGrath, Guy...... OA45.01 ...... PS03.84, PS03.13, OA22.02, OA33.02, McHugh, Mark...... PS01.21 ...... OA33.03, OA33.04, OA33.05, OA33.06, McIntyre, Jennifer...... PS01.79 ...... OA38.01, OA38.02, OA38.03, McKay, L...... OA34.04 ...... OA38.04, OA38.05, OA38.06, OA48.01, McKenzie, Catherine...... PS03.34, OA41.03 ...... OA48.06 McKenzie, John...... PS02.12 Nimsuphan, Burin...... PS02.36 McLoughlin, Philip...... OA48.07 Nisbet, Alasdair J...... PS02.49, PS03.61 McMahan, Christopher...... OA01.01, PS03.70 Nixon, Samantha...... OA50.06 McManus, Concepta...... PS01.71 Nöckler, Karsten...... OA30.04 Noh, Jinhyeong...... PS01.47, PS02.38, PS03.44 N Normile, Dorothy...... OA01.03, OA01.05 Norris, Jamie...... OA37.04, OA48.06 Nadeem, Raaziya...... OA50.03 Novobilský, Adam.PS02.04, PS03.45, PS02.43 Nader, Sara...... OA04.02 Nurcahyo, Wisnu...... OA27.03, OA27.05 Nagamori, Yoko...... OA21.05 Nwosu, Chukwunyere...... PS02.86 Nagar, Gaurav...... OA49.07 Nzeakor, Terry...... PS02.86 Nagata, Walter Bertequini...... OA09.03 Nzekwe, Sandra...... PS02.86 Naik, V.C.S...... OA46.07 Nakaguchi, Victor....PS01.08, PS02.14, OA07.06 O Nakatani, Matheus...... PS01.02 Napoli, Ettore...... OA21.01 O’Neill, Emma...... PS02.59 Naranjo Lucena, Amalia...... OA07.02, OA45.01 O’Shaughnessy, James...... PS02.59 Nascimento, Cristiano...... PS01.54, PS01.59, Obeta, Sylvester...... OA29.02, PS03.12 ...... PS02.82, PS03.55 Obi, Francis...... PS02.86 Nascimento, Isabela...... PS02.39 O’Connor, Jack...... OA49.06 Nascimento, Janilene...... OA23.03 Odhiambo, Brenda...... OA12.06 Naseer, Muhammad Usman...... OA49.05 Odongo, David O...... OA29.04 Nasr, Soad...... OA37.01 Ogo, Ndudim...... OA29.02 Natala, Joseph...... OA29.02 Ojeda-Chi, Melina M...... PS01.73, PS02.62, Nautrup Olsen, Susanne...... PS02.08 ...... OA39.03 Navaluna, Edelmar...... OA04.01 Ojo, Kayode K...... OA04.01 Navarro, Christelle....PS01.01, PS01.34, PS01.60, Okal, Michael...... OA12.06 ...... PS03.02, PS03.08, PS03.63, Okino, Cintia Hiromi...... PS03.51 ...... PS03.75, OA39.05 Okorie, Victor...... PS03.12 Nayduch, Dana...... PS02.71 Olafson, Pia...... OA34.04, OA43.02 Ndao, Momar...... OA46.03 Olayemi, Olutayo-Daniels...... PS01.55, PS02.48, Neelakshi, Deka...... PS02.87 ...... PS01.56 Nemer, Nabil...... OA42.02 Oleson, Jacob...... PS01.69 Neola, Benedetto...... OA05.05, OA26.01 Olhagaray Torres, Maria Ernestina...... PS02.31,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 341 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

...... PS02.32 Park, Yeojin...... PS01.47, PS02.38, PS03.44 Oliva, Gaetano...... OA23.02, OA23.05, PL03.01 Parker, Grant...... PS03.39 Oliveira, Ana Paula...... PS03.64 Parrish, Molly...... PS01.69 Oliveira, Bergson...... PS02.83 Parrish, Rudolph...... OA06.01 Oliveira, Bruno...... OA09.03 Paskewitz, Susan...... OA11.05, OA44.04 Oliveira, Gabriela...... PS02.82 Pastor, Adriana...... PS03.41 O’Neill, Emma...... PS02.59 Paterson, Steve...... OA24.07 O’Shaughnessy, James...... PS02.59 Paterson, Tara...... PS01.66, OA05.03, OA06.08, Obeta, Sylvester...... OA29.02, PS03.12 ...... OA27.04 Obi, Francis...... PS02.86 Patricio Monteiro, Jomar...... PS02.24, PS02.63 O’Connor, Jack...... OA49.06 Patsanza, Gloria...... PS02.64 Odhiambo, Brenda...... OA12.06 Patullo, Cailey...... OA41.01 Odongo, David O...... OA29.04 Pawlak, Jesse...... PS01.17 Ogo, Ndudim...... OA29.02 Payne, Patricia...... OA10.06 Ojeda-Chi, Melina M...... PS01.73, PS02.62, Payton, Mark...... OA08.07, OA21.05 ...... OA39.03 Paz, Jessica Priscila da...... PS02.27, PS02.84 Ojo, Kayode K...... OA04.01 Peachey, Laura...... OA43.06 Okal, Michael...... OA12.06 Pearl, David...... OA20.02 Okino, Cintia Hiromi...... PS03.51 Pelissoni, Luis...... PS01.54, PS01.59, PS02.82, Okorie, Victor...... PS03.12 ...... PS03.55 Olafson, Pia...... OA34.04, OA43.02 Pelletier, Claire...... PS01.03 Olayemi, Olutayo-Daniels...... PS01.55, PS02.48, Peña-Espinoza, Miguel...... OA28.03 ...... PS01.56 Penner, Gregory...... OA36.02 Oleson, Jacob...... PS01.69 Pennisi, Maria-Grazia...... OA18.01 Olhagaray Torres, Maria Ernestina...... PS02.31, Penzhorn, Banie...... OA11.02, PS01.62 ...... PS02.32 Pepe, Paola...... OA20.03 Oliva, Gaetano...... OA23.02, OA23.05, PL03.01 Peregrine, Andrew.PS02.20, PS02.69 OA20.02, Oliveira, Ana Paula...... PS03.64 ...... PS03.22 Oliveira, Bergson...... PS02.83 Pereira de Freitas, Edilson...... PS02.63 Oliveira, Bruno...... OA09.03 Pereira de Oliveira, Gilson...... PS01.38 Oliveira, Gabriela...... PS02.82 Peres, Pedro...... OA09.03 Pérez, José...... OA45.01 P Pérez de León, Adalberto...... OA03.04, ...... OA08.06, OA49.02, OA49.03, OA49.04, Pacheco, Poliana Araújo...... PS01.74, PS01.22, PS02.30, PS02.62, PS02.78, PL04.03, PS03.59 PS01.48, PS02.23, PS02.27, PS02.45, PS02.84, Pérez Sarasqueta, Ana Laura.PS02.31, PS02.32 ...... PS03.28, PS03.50, PS03.56 Pérez Tort, Gabriela...... PS02.53, PS02.54, Pachnicke, Stefan...... OA11.04 ...... OA21.04 Pacifico, Laura...... OA05.05, OA26.01, OA33.01 Pérez-Fonseca, Agustin...... PS03.14 Pagac, Alexandra...... OA32.04 Pérez-Hernández, Tara...... PS02.49, PS03.61 Page, Greg...... PS03.73 Peters, Sunday...... PS02.35 Paiva, Jonimar...... PS03.80 Petersen, Christine.PS01.69, OA18.04, OA23.01 Paller, Vachel Gay Noreen.....OA47.08, PS02.88 Petersson, Katherine...... PS03.23, OA28.02 Palmeira, Bruna...... OA46.01 Pettersson, Emelie...... OA22.08 Palmer, Zachary...... PS01.69 Philippi-Taylor, Aimee...... OA36.01 Panarese, Rossella...... OA09.04, OA21.01 Phillips, Pamela...... PS02.30 Pantchev, Nikola...... PS02.51, OA26.02 Phiri, Isaac K...... PS01.62 Papadopoulos, Elias...... PS01.80, PS03.63, Phoosangwalthong, Pornkamol...... PS01.84 ...... PS03.75, PS03.83 Piantedosi, Diego.OA05.05, OA26.01, OA33.01 Paras, Kelsey...... OA12.03, PS02.72, OA32.01 Pickens, Victoria...... PS03.19 Paraud, Carine...... PS02.11 Piedrafita, David Michael...... OA07.01 Park, Brandon...... OA31.02 Pierson, Christopher...... PS02.58 Park, Doyeon...... PS02.19, PS03.20 Pike, Chelsea...... PS03.59 Pilfold, Nicholas...... OA25.07

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 342 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Pilotte, Nils...... OA06.02 Rao, Arundhati...... OA36.04 Pinn-Woodcock, Toby...... OA41.02 Rashid, Harun...... OA41.04 Pinto, Nathalie Silva...... PS01.16 Rashid, Imran...... PS01.77, OA46.06 Plantard, Olivier...... OA13.04 Rathinasamy, Vignesh...... OA17.04 Plattner, Jacob J...... OA36.07 Rauh, Renate...... PS03.83, OA36.05 Poissant, Jocelyn. OA38.05, OA48.07, PS03.74 Raul Correia de Vasconcelos, Fernando...... Poitevin, Manoel...... OA39.01 ...... PS02.24 Pol, Glòria...... OA23.06 Rauta de Avelar, Barbara...... PS03.02, OA39.05 Polack, Bruno...... PS01.03, PS01.68, OA25.04, Ravinet, Nadine....PS01.76, OA02.04, OA02.05, ...... OA25.05 ...... OA12.01 Pollmeier, Matthias...... OA11.03 Read, Emma...... OA15.05 Pollock, Colleen...... PS02.76, OA36.02 Reck, Jose...... OA39.01 Pombi, Marco...... PS03.38 Reckziegel,...... PS01.02 Pomroy, Bill...... OA12.02 Recta, Virginia...... OA36.01 Pongracz, Jodie...... PS03.32 Redman, Elisabeth.. PS01.17, PS02.76, PL02.04, Poole, William...... OA17.04 ...... OA19.02, OA24.06, OA36.02 Portillo, Laura...... PS02.80 Redman, Libby...... PS03.74, OA41.01, OA46.01 Powell-Coffman, JoAnne...... PS01.21 Reed, Jess...... PS03.23 Power, Cormac...... PS02.59 Rego, Ryan O. M...... OA27.02 Prakash, Manu...... PS02.19, PS03.20 Rehbein, Steffen...... PS02.03, PS03.29 Prevot, Francoise... PS01.09, PS02.74, OA46.02 Rehbein, Steffen...... PS03.83, OA36.05 Price, Dan R.G...... PS02.49, PS03.61 Rehman, Tauseef...... OA50.05 Prichard, Roger...... PS01.62, OA01.04 Reichard, Mason.....OA13.03, PS02.01, OA29.05 Prieto, Noemi...... PS01.66 Reid, Gavin...... OA19.06 Pritsch, Izanara...... OA50.04 Reinemeyer, Craig.OA14.04, OA38.01, OA38.02 Priyavoravong, Patcharin...... PS01.49 Rejman, Evelin...... OA14.05 Pruzzo, Cesar...... OA35.01 Rekik, Mourad...... PS01.33 Puget, Nollan...... PS03.15 Ren, Qiaoyun...... OA03.05 Pulaski, Cassan...... OA01.01, OA01.04, PL02.03 Reyes-Novelo, Enrique...... PS01.73 Pürkel, Simone...... PS02.51 Rhimäkii, Miia...... OA33.05 Ribeiro, Enilson Geraldo...... PS02.73, PS03.51 Q Ribeiro, Jessica Carolina...... PS01.22, PS01.74, ...... PS02.73, PS02.23, PS03.28, PS03.56 Qamar, Muhammad Fiaz...... PS01.85, PS02.56, Ribeiro Campos, Difrey...... PS03.18, PS03.82 ...... PS03.71, OA28.04, OA32.05, OA50.07 Rich, Brian...... OA49.03 Qamar, Warda...... PS01.85, PS02.56, PS03.71 Richards, Grant...... OA02.06 Qu, Zhiqiang...... OA03.05 Richter, Rüdiger...... PS03.68 Quaglio, Francesco...... OA09.04 Riet Correa, Franklin...... PS03.40 Qudoos, Abdul...... OA50.03 Rinaldi, Laura...... OA12.01, PS02.79, OA20.03, Queiroz, Camila...... PL02.04, PS01.17 ...... PS03.07, PS03.31 Quenet, Yann...... OA02.04, OA02.05 Rios de Alvarez, Leyla...... PS01.24, PS01.23 Quilis, Dalmo...... PS02.52, PS03.64 Risco-Castillo, Veronica...... PS01.68 Qurollo, Barbara...... OA05.04 Rispe, Claude...... OA13.04 Ritchey, Jerry...... OA11.01 Ritchie, Debra...... OA10.06 R Rivera-Jacinto, Marco...... PS01.51 Riveros, José Mariano...... PS01.10 Rabinowitz, Peter M...... KL01.03 Rizwan HM,...... PS03.48, OA50.03 Raja, Thamayanthi Nada...... OA40.01 Rjeibi, Mohamed Ridha...... PS01.33 Rajagopalan, Srinivasan...... OA10.07 Robertson, Alan...... PS01.15, PS01.21, OA01.06, Rakitin, Andrei...... OA05.01 ...... OA19.04, OA19.05, OA22.04, OA26.06 Ramünke, Sabrina...... OA12.04 Robinson, Mark...... OA35.05 Rani, Puteri Azaziah Megat Abd...... OA27.03, Robinson, Samuel...... OA50.06 ...... OA27.05 Rochon, Kateryn...... OA34.06

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 343 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Rodrigueiro, Tatyana Sacchi Carmona.PS02.50 Sagar, Rashmi...... PS01.58 Rodrigues, Leandro.PS01.48, PS01.74, PS02.23, Sagel, Agustin...... PS02.30 PS02.27, PS02.44, PS02.45, PS03.49, PS03.50 Sager, Heinz...... OA44.01 Rodrigues, Paulo...... PS01.20 Sagües, Federica...... PS01.83, PS02.47, PS03.57 Rodrigues, Suzete...... PS01.20 Sagynbek, Shermatov...... PS02.26 Rodrigues, Vinícius...... PS01.02 Sahraoui, Lynda...... OA25.05 Rodrigues da Silva, Felipe...... PS02.63 Said, Asraf...... OA08.08 Rodrigues, VD...... OA34.02 Said, Yousra...... PS01.33 Rodriguez, Jessica Y...... OA47.05 Sajid, Muhammad Sohail...... PS03.48, OA17.06, Rodriguez, Maria Ines Genovese...... PS03.62 ...... OA50.03 Rodríguez, Antonio...... PS01.10, PS02.80 Sakamoto, Claudio A Massamitsu...... PS02.84 Rodríguez-Durán, Arlex...... PS02.78 Sakamoto, Kaori...... PS03.72 Rodríguez-Ulloa, Claudia...... PS01.51 Sakita, Gabriel Z...... PS02.14 Rodriguez-Vivas, Roger Ivan..PS01.27, PS01.73, Sakong, Bellonah...... PS03.16 ...... PS02.62, OA39.03 Salazar-Ulloa, Martha...... OA22.05 Roeber, Florian...... PS03.78 Saldaña, Jenny...... PS03.30 Roellig, Dawn...... OA25.02 Saleem, Mushtaq A...... PS01.77 Roemmele, Eric...... OA33.02 Saleh, Meriam...... OA03.01, OA03.02, OA03.03, Rogers, Laura...... OA26.06 ...... PS02.42 Rogers, Sean...... OA24.05 Salim, Bashir...... OA08.08, OA17.02 Rojas, Diana...... PS01.23, PS01.24 Salles, Bruna...... OA41.01 Rolfe, Peter...... OA35.04 Sallovitz, Juan...... PS01.82, PS01.83 Romagnoli, Emiliana...... PS01.71 Salvador, Davi...... PS02.52 Roman, Constantin...... OA13.05 Sampaio, Ana...... PS01.71 Romanos, Lola...... PS02.74 Sampaio, Paulo Henrique Selbmann...... PS01.57, Romero-Ramirez, Alessandra...... OA29.01 ...... PS02.50, PS03.62, PS03.65, OA32.02 Roncoroni, Cristina...... OA33.01 Sanabria, Rodrigo...... OA35.01 Rosado Aguilar, José Alberto...... PS01.27 Sanchis, Jaime...... OA35.01 Rosenthal, Benjamin...... PS03.32 Sandalo, Kristina Andrea...... OA39.02 Rossi, Claudio...... OA23.03 Sanders, John...... OA50.02 Rotolo, Jessica...... OA37.05, OA37.06 Sanders, Justin...... OA15.03 Roux, Cecile...... PS03.69 Sandey, Maninder...... PS01.39 Rozane, Danilo Eduardo...... PS03.62 Sangaran, Arumugam...... OA21.04, PS02.87 Ruano, Ricardo...... OA26.02 Sanisuriwong, Jitra...... PS01.14 Rudinsky, Adam...... OA16.02 Santana, Bruna Nicoleti...... OA09.03, PS02.39 Rufrano, Domenico...... OA33.01 Santaniello, Mirella...... PS03.07 Rugg, Doug...... OA01.02 Santarém, Vamilton...... PS02.27 Rugg, Jady...... PS02.01 Santin, Monica...... OA25.03, OA42.06 Rumschlag, Anthony...... OA44.02 Santoro, Mario...... PS02.15 Rus, Luciana Cătălina...... OA21.02 Santos, Bianca Martins dos...... OA37.03 Rustas, Bengt-Ove...... PS02.04 Santos, Carolina...... OA50.04 Ruttkowski, Baerbel...... OA04.01, OA04.03 Santos, Edna Michelly...... OA23.03 Ryabova, Nadezhda...... PS02.68 Santos, Eveline Henrique dos...... PS02.84 Ryan, William...... OA44.02 Santos, Isabella...... PS02.22, PS02.52 Santos, Taminis...... PS01.59 S Santos De Miranda, Mariana...... PS02.44 Saporitti, Tatiana...... PS01.75 S Jung, Tom...... PS03.32 Saraiva Bresciani, Katia Denise...... PS01.38 S. Leifsson, Páll...... PS02.08 Saravia, Anderson...... PS02.32, PS03.40 Saarma, Urmas...... PS02.15 Sargison, Neil D...... PS01.77, PS03.53, PS01.79, Sabatini, Gustavo...... PS01.54 ...... OA24.04, PS03.24, OA31.01, OA46.06 Saeed, Eman Magzoub...... PS01.29 Sarnelli, Paolo...... PS02.15 Saegerman, Claude...... PS03.47 Sartor, Debora...... PS01.54, PS01.59, PS02.82, ...... PS03.55

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 344 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Sarzen, Marcel...... OA44.02 Sharma, Sonu...... OA49.07 Saucier, Jill...... PS01.69, OA18.04 Sharpton, Thomas...... OA15.03 Sauermann, Christian...... OA07.03, OA38.01, Shaukat, Ayaz...... PS01.77 ...... OA38.02, OA38.03, OA38.04, OA38.06 Shaw, Nicolette...... PS03.74, OA46.01 Saumell, Carlos...... PS01.83, PS02.47, PS03.57 Shahzad, Muhammad...... OA09.07 Saunders, Lori...... OA06.02 Sheikh, Razan...... OA26.05 Savinov, Sergey...... OA22.03 Shen, Hui...... OA03.05 Scala, Antonio...... PS02.15 Sheng, Qicong...... OA19.01 Scandrett, Brad...... PS03.32 Sheng, Zhao-An...... OA35.06, OA45.05 Scare, Jessica...... PS03.84, OA48.01, OA48.06 Shepherd, Taylor...... OA33.03 Schaarschmidt-Kiener, Daniel...... OA11.04 Shigaki de Matos, Lucas Vinicius...... PS01.38 Schaefer, John...... PS03.66 Shrake, Jessica...... PS02.57 Schaper, Roland...... OA06.08, OA11.04, PS01.84 Shrestha, Aruna. OA04.01, OA04.03, OA04.04 Schetters, Theo...... OA39.04 Shryock, Jeffrey...... PS02.03 Schexnayder, S...... OA34.04 Shury, Todd...... OA48.07 Schlechte, Kristie...... OA08.06 Sibeko-Matjila, Kgomotso...... PS02.64 Schmaeschke, Ronald...... PS03.68 Siebert, Susanne...... PS01.84 Schmidt, Olaf...... OA49.06 Silas, Haley...... PS01.17 Schneider, Derek...... OA35.02 Silva, Carolina Rocha.PS01.16, PS01.25, PS01.26 Schnyder, Manuela...... OA06.07, OA11.04, Silva, Diogo...... OA18.06 ...... OA26.04, PS03.09, PS03.67 Silva, Ellen...... PS01.16 Schofs, Laureano...... OA39.08 Silva, Liliana M.R...... PS01.37 Scholze, Germano...... PS03.49 Silva, Marta...... OA17.04 Schroeder, Christina...... OA50.06 Silva, Naylene Carvalho Sales...... PS01.16 Schunack, Bettina...... OA11.03, OA18.05 Silva, Valmir...... OA23.04 Schuppisser, Carole...... OA06.07 Silva, Carolina...... PS03.55 Schurer, Janna...... OA20.01 Silva, Fabiano...... OA13.01 Schwartz, Andy...... PS03.59, OA49.02 Silva, Maria R. L...... PS01.26 Scimeca, Ruth...... OA13.03, OA29.05 Silva, Naylene...... PS01.59 Scoggin, Kirsten...... OA48.06 Silva, Winny...... OA23.03 Scorza, Breanna...... PS01.69, OA18.04, OA23.01 Silva Pereira, Sara...... OA29.01 Scott, Fabio...... PS01.54, PS02.82 Silva Vieira, Luiz...... PS02.24 Scott, Ian...... OA12.02 Silveira Neto, Luiz...... PS01.38 Ségalen, Coralie...... PS01.60, PS03.63 Sim, Sheina...... PS02.30 Seguin, M. Alexis...... OA42.03 Simonsen, Henrik Toft...... OA28.01, OA28.03 Self, Stella...... OA01.01, PS03.70 Sindhu, Zia ud Din...... OA49.05 Sellal, Eric...... PS01.03 Singh, Balbir...... PS02.88, OA40.01 Senior, Eleanor...... OA22.06 Singla, Lachhman...... OA42.05 Serrano, Celeste...... PS02.55, PS02.79 Široký, Pavel...... PS02.43 Servonnet, Marielle...... PS02.40 Sjölund, Marie...... OA22.08 Sgroi, Giovanni...... OA05.05, PS02.15, OA26.01, SK, Kiran...... OA08.04 ...... OA33.01 Skvarla, Michael...... OA09.01 Shaapan, Raafat...... OA08.08 Slapeta, Jan...... OA15.02, OA30.03, OA44.03 Shabangu, Angy...... OA11.02 Slaughter, Jana...... OA11.01 Shabbir, Muhammad Zubair...... OA46.06 Slusarewicz, Paul.OA33.02, OA48.01, OA48.06 Shaffer, Carrie...... OA48.01 Smit, Nico J...... PS03.37, OA09.06, OA37.02 Shahid, Zohaib...... PS01.85, PS03.71, PS02.56 Smith, Emily...... OA36.01 Shahzad, Kashif...... PS03.53 Smith, Vicki...... OA10.06 Shakya, Mukesh...... OA49.07 Smits, Debora...... OA46.05 Shamsi, Shokoofeh...... OA15.02 Snigdha, Kancharana...... OA39.07 Sharif, Shukri...... OA13.06, PS03.58 Snowden, Karen F...... OA47.05 Sharma, Anil Kumar...... OA49.07 Snyder, Ryan...... , PS01.53PS03.73, OA37.05, Sharma, Nidhi...... PS01.72 ...... OA37.06 Sharma, Rajnish..... OA20.01, OA25.07, PS03.32 Soares, Alexandra Martins Santos...... PS01.25

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 345 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Soares, Nilce Maria...... OA32.02 T Soares, Rodrigo Martins...... PS03.62 Soares, Vando...... PS02.22, PS03.64 Tagwireyi, Whatmore...... OA25.06, OA47.01 Soares, Alexandra M. S...... PS01.26 Talactac, Melbourne...... OA03.06 Solano-Gallego, Laia...... PS02.65, OA18.03, Talley, Justin...... OA11.01, PS02.06 ...... PS03.85, PL03.01 Tamkeen, Rabia...... OA50.07 Sorge, Colleen...... OA21.06 Tamponi, Claudia...... PS02.15 Sonzogni-Desautels, Karine...... OA46.03 Tan, Do Yew...... OA27.03, OA27.05 Sotera, Salvo...... OA23.02 Tanaka, Tetsuya...... OA03.06, OA39.02 Sotillo, Javier...... OA22.01 Tao, Sha...... PS02.02 Sotiraki, Smaragda...... PS03.31 Tarallo, Viviana Domenica...... OA18.05 Soufy, Hamdy...... OA37.01 Tarpalechee, Layna...... OA11.01 Souil, Elise...... PS02.74 Tasker, Séverine...... OA44.05 Sousa, Isabella Chaves...... PS01.59, PS03.55, Taubert, Anja...... PS01.67, PS01.37, PS02.37, ...... PS03.60 ...... PS03.03, PS03.04 Souza, Celeste...... PS01.64, OA23.04 Tavares, Caio Pavão...... PS03.60 Souza, Pedro Filho Noronha...... PS01.25 Tavendale, Michael...... OA41.03, PS03.34 Spangler, Dawn...... OA06.06 Tavengwa, Michael...... OA47.01 Speiser Fontaine, Christelle...... OA23.05 Taweethavonsawat, Piyanan...... OA27.03, Sperling, Daniel...... OA04.04 ...... OA27.05, PS01.49 Springer, Andrea...... OA40.02 Taylor, D. B...... OA34.04 Srivastava, Sharad...... OA49.07 Taylor, David...... PS02.33 Starkey, Lindsay..... PS01.39, OA06.03, PS02.19, Tedesco, Perla...... OA09.04 ...... PS03.20 Teel, Pete...... PS01.44, OA49.01, OA49.02, Stasiuk, Susan...... PS01.72 ...... OA49.03, OA49.04 Steffan, Pedro...... OA39.08 Teixeira, Marcel...... PS01.70, PS02.24, PS02.63 Steuer, Ashley...... PS02.25, PS03.84, OA33.03, Teixeira, Ramiro...... PS03.30 ...... OA33.04, OA48.01, OA48.06 Teixeira, Weslen...... OA09.03 Stewart, John...... OA33.04 Temeyer, Kevin...... OA08.06 Stockdale Walden, Heather...... OA06.03 Ternent, Mark...... OA09.02 Stofanak, Heather...... OA15.01 Thamsborg, Stig.... OA02.01, OA16.01, OA28.01, Stomoxys Genome Consortium,...... OA43.02 ...... OA28.03 Storey, Bob...... OA31.02 Thany, Steeve...... OA13.04 Stout, Alison...... OA43.01 Thekisoe, Oriel...... OA09.06, PS03.46 Stowe, Sarah...... PS01.66 Thiengo, Silvana...... PS01.20 Strube, Christina...... PS03.67, OA40.02 Thomas, Arme...... PS03.38 Stuart, Peter...... OA47.07 Thomas, Brittany...... OA05.04 Su, Chunlie...... OA30.01 Thomas, Donald.... PS02.46, PS03.59, OA49.03 Suarez, Carlos...... OA17.04 Thomas, Emmanuel...... PS01.30 Sudlovenick, Enooyaq...... OA25.07 Thomas, Jennifer...... PS02.01 Sugiyama, Hiromu...... OA45.04 Thomas, Myriam... PS01.03, OA25.04, OA25.05 Sumption, Derek...... PS01.62 Thompson, Alec...... PS01.04, PS02.70, PS03.33 Sun, Hongchao...... PS01.40 Thompson, Belinda...... OA41.02, OA43.01 Sun, Miao-Miao...... OA45.04 Thompson, Peter...... PS03.32 Sundstrom, Kellee...... OA03.01, OA03.02, Thonney , Michael...... PS03.23 ...... OA03.03, PS02.42, PS03.10 Thontiravong, Aunyaratana...... PS01.14 Sundstrom, Kellee...... OA26.03 Thyssen, Patricia...... OA13.01 Sutra, Jean-Francois...... PS02.74 Tiawsirisup, Sonthaya...... PS01.14 Suzuki, Celso Tetsuo Nagase...... OA37.03 Tidwell, Jason...... OA03.04 Swain, Lana...... PS01.17 Tiemi Akashi, Karen...... PS02.44 Swart , W.A.J.M...... OA46.05 Tipton, James G...... PS01.15 Swiger, Sonja...... OA34.03, OA49.01 Tirloni, Lucas...... PS01.13, OA08.01, OA08.02 Symeonidou, Isaia...... PS01.80, PS03.63 Tiwananthakorn, Saruda...... OA27.03, OA27.05, Szlosek, Donald...... OA10.02 ...... PS02.36

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 346 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

To, Tran...... PS03.25, PS03.35 Valera, Fred Gio Rean...... OA39.02 Toepp, Angela...... PS01.69, OA23.01 Valladares, María Martínez...... OA28.01 Toledo, Luciandra Macedo...... PS03.51 Vallée, Isabelle...... OA25.04, OA25.05 Tolleson, Douglas...... OA49.03 van As, Johann...... OA09.06 Tollett, Christina...... OA48.07 Van As, Michelle...... OA09.06 Tomberlin, Jeffery...... OA49.01 van den Heuvel, Louise...... OA27.01 Tong, Kenneth Boon Yew...... OA27.03, OA27.05 Van Dijk, Jan...... OA19.02 Tontong, Siriphon...... PS02.36 van Engelen, Erik...... OA46.05 Torgerson, Paul...... OA02.01 Van Galen, Gaby...... OA15.02 Torres Islas, Juan Agustin...... OA39.03 Van Mael, Eva...... PS03.78 Torres-Acosta, Felipe...... PS01.16 Van Mol, Wouter...... PS01.81, OA39.06 Toscano, João...... PS02.22 van Wyk, Ilana...... PS01.42 Toscano, Valerio...... PS02.15 Van Why, Kyle...... OA30.01 Toubate, Berthine...... OA13.02 Vande Velde, Fiona...... PS03.78 Touchard, Axel...... OA50.06 Vanhecke, Marieke...... PS01.81, OA39.06 Tracey, Alan...... PS03.24 VanHoy, Grace...... PS02.72 Traub, Rebecca...... OA21.04 Varcasia, Antonio...... PS02.15 Trerotoli, Paolo...... OA18.05 Vargas-Duarte, Jimmy Jolman...... OA50.03 Trinidad-Martinez, Iris...... OA39.03 Varuzza, Paolo...... PS02.15 Trinidad-Martínez, Imelda...... PS01.73 Vasconcelos Meireles, Marcelo...... PS01.38 Tritten, Lucienne....PL01.03, OA26.04, PS03.09 Vassilev, George...... OA47.01 Trout Fryxell, R. T...... OA34.04 Vatta, Adriano...... PS02.01 Truter, Marliese...... PS03.37 Vázquez Vega, Cecilia Marina...... OA32.06 Tsai, Siu...... PS01.71, PS03.56 Velasquez, Zahady. PS01.37, PS02.37, PS03.03, Tsai, Yi-Lun...... OA08.05, OA27.03, OA27.05 ...... PS03.04 Tsuji, Naotoshi...... OA03.06 Velthuis, A.G.J...... OA46.05 Tucci, Edna Clara...... OA32.02 Veneziano, Vincenzo...... OA05.05, PS02.15, Tunceli, Kaan...... OA01.05, OA10.07 ...... OA33.01 Tuo, Wenbin...... OA07.05 Venkatesan, Abhinaya...... PS03.26 Turnbull, Frank...... OA19.03 Venturina, Virginia...... OA27.03, OA27.05 Tursi, Angelo...... OA09.04 Venu, Ravipati...... OA46.07 Tydén, Eva...... OA33.05, OA48.02, OA48.03, Vercesi Filho, Aníbal...... PS03.51 ...... OA48.04 Vercesi Filho, Aníbal...... PS02.73 Tyrell, Phyllis...... PS01.69 Vercruysse, Jozef...... OA02.01 Tyrrell, Phyllis...... OA05.03, OA18.04, OA27.04 Veríssimo, Cecília José...... PS01.22, PS01.48, Tzelos, Tom...... OA33.06 PS01.74, PS02.23, PS02.27, PS02.44, PS02.45, PS02.73, PS02.84, PS03.28, PS03.49, ...... U ...... PS03.50, PS03.51 Verma, Ankita...... OA49.07 Uehlinger, Fabienne...... PS01.17, PS02.76, Verma, Saurabh...... PS01.21, OA01.06, OA19.04, ...... OA24.06, OA36.02 ...... OA19.05, OA22.04 Uggla, Arvid...... OA33.05 Verocai, Guilherme...... OA16.03, OA21.03 Vetter, Irina...... OA50.06 Vettorato, Luis...... PS01.30 V Vialard, Fiorella...... PS03.41 Vidyashankar, Anand...... OA31.02, OA32.01 V, Manjunathachar...... OA49.07 Vieira, Douglas...... OA50.04 Va, Florence...... PS01.03 Vieyra-Gómez, Natalia...... PS01.05 Vaala, Wendy...... PS03.11, OA43.03 Villar, David...... PS02.78 Valadares, Diogo...... OA23.01 Villar, Margarita...... OA27.02 Valderas, Elora...... OA36.08 Villar-Argaiz, David...... PS01.67 Valderas García, Elora...... OA41.05 Villegas–Perez, Sandra L...... PS02.62 Valdez, Norma...... PS01.66 Vinay, S...... PS01.12 Vale, Tássia Lopes.. PS01.59, PS03.55, PS03.60 Vincent, Emily...... OA16.02

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 347 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

Vineer, Hannah...... PS01.79 Wedy, Bruna...... PS01.22, PS01.74, PS02.23, Vinueza Veloz, Pamela...... PS03.78 ...... PS03.28, PS03.56 Virkel, Guillermo....PS02.83, PS02.85, PS03.76, Wei, Nana...... OA17.01 ...... OA39.08 Weisbecker, Jean-Louis...... PS01.09 Visser, Martin...... OA36.05, PS02.03 Welch, John...... PS02.30 Vitale, Fabrizio...... OA23.02 Wells, Bridgette...... PS02.69 Vito, Gabriela...... PS01.59, PS03.55 Wentzel, Jeanette...... PS01.42 Viviani, Paula...... PS03.76 Werdal, Emily...... OA21.06 von Samson-Himmelstjerna, Georg...... OA11.04, Werre, Stephen R...... PS01.62 ...... OA12.04, OA22.02, PS01.76, OA26.02, Wetherly, Patricia...... OA26.05 ...... PS02.79, PS03.13, PS03.31 Weyl, Olaf LF...... PS03.37 von Son-de Fernex, Elke...... PS01.28, OA12.04, Wheeler, Nic...... PS03.21, PS03.35 ...... OA22.05, OA50.01 Wheeler, Nicolas...... PS01.18, PS03.25 Voorhis, Wesley C. Van...... OA04.01, PL01.02 Wheling, Libby...... OA33.02 Voskuilen, Timo...... OA27.01 White, Holly...... PS02.61 VVS, Suryanarayana...... OA40.07 Whitelaw, Amelia...... PS01.17 Whitman, Grant R...... OA04.01 W Whittaker, Gary R...... OA43.01 Widmer, Giovanni...... PS01.38 Waap, Helga...... PS01.76 Wiefel, Thea...... OA36.05 Wachowski, Martha...... OA01.02 Wielgus, Kayla...... OA33.02 Waghorn, Tania...... OA07.03, PS03.34, OA41.03 Wijnveld, Michiel...... OA15.04 Wagner, Brent...... OA25.07, PS03.32, OA40.06 Williams, Andrew Richard.....OA28.01, OA28.03 Waindok, Patrick...... OA40.02 Williams, Diana...... OA24.07, PS03.54 Walden, Heather..... PS01.61, OA06.05, OA15.05 Williams, Paul...... PS01.21 Walkden-Brown, Stephen... OA07.04, OA14.03, Williams, Steven...... OA06.02 ...... PS02.29, PS03.39 Williamson, Nicholas...... OA19.06 Walker, Andrew...... OA50.06 Will, Hermann...... PS02.51 Wall, Richard...... OA44.05, PS03.42 Wilson, Elizabeth...... PS01.66 Wallgren, Per...... OA22.08 Wilson, Geneva...... PS01.69 Walshe, Nicola...... OA38.07 Wilson, Georgette...... OA13.03 Wang, Chunqun...... OA24.08 Wiśniewski, Marcin...... PS02.66 Wang, Hailun...... PS03.83, OA36.05 Wit, Janneke...... PL02.04, PS03.74, OA24.05, Wang, Hsiao-Hsuan...... OA49.02, PS01.44 ...... OA41.01 Wang, Jianbin...... OA22.02, OA48.06 Witola, William...... OA04.02 Wang, Jiangwei...... OA27.03, OA27.05 Wohltjen, Megan.....OA11.01, OA13.03, OA26.03, Wang, Jin-lei...... OA04.06 ...... PS03.10 Wang, Lijie...... PS03.17 Wolday, Mengisteab...... OA22.04 Wang, Qingcan...... OA04.05 Wolfe, Alan...... OA45.01 Wang, Shuai...... PS02.17 Workentine, Matthew...... OA24.05, OA46.01, Wang, Tao...... OA19.06 ...... PS03.74, PS03.26 Wang, Tong...... OA41.01 Wright, Gavin...... OA29.01 Wang, Yuanfei...... OA25.02 Wright, Harry...... PS02.49, PS03.61 Wang, Yujian...... OA17.01 Wu, Joy...... OA40.06 Wang, Zhu...... PS02.21 Wu, Shubiao...... OA14.03 Ward, Erica...... PS01.17 Wu, Zegong...... OA03.05 Wasmuth, James...... OA24.05, PS03.26 Wulcan, Judit...... OA21.03 Watanabe, Malaika.OA21.04, OA27.03, OA27.05 Wyckoff, Seth...... PS03.33 Watson, Wes...... PS01.32 Wyrosdick, Heidi...... PS03.66 Wayadande, Astri...... PS03.19 Wysmołek, Magdalena Elżbieta...... PS02.66 Weber, Corinna...... OA11.04 Wechtaisong, Wittawat...... OA08.05 Wedlon, Polly...... PS02.09

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 348 63rd Annual Livestock Insect Workers Conference AUTHOR INDEX

X ...... OA29.03 Zhang, Lei...... OA17.05 Xiao, Lihua...... OA25.02 Zhang, Shaohua...... PS02.17, PS03.17 Xu, Xiaofeng...... OA03.05 Zhang, Wei-Yu...... OA45.05 Xu, Yumin...... PS02.02 Zhang, Xiao-Xuan...... OA35.06 Xu, Zhengmao...... OA17.01 Zhang, Xuejin...... OA04.02 Xuan, Xuenan...... PS01.35, PS01.36 Zhang, Yong-Kang...... OA36.07 Zhang, Hui...... OA09.07 Zhao, Xiongce...... OA36.01 Y Zheng, Wen-Bin...... OA35.06, OA45.05 Zheng, Yadong...... PS03.17 Yabsley, Michael...... PS01.04, OA01.01, PS02.70, Zhijuan, Yin...... OA27.03, OA27.05 ...... PS03.33, PS03.70 Zhou, Cheng-Yan...... OA45.04 Yam, Jerald...... OA17.03 Zhou, Ershun...... PS03.04 Yamasaki, Fumiaki...... OA40.05 Zhou, Jinlin...... OA04.05, OA17.01, OA29.03 Yamassaki, Fabio...... OA50.04 Zhou, Yasheen...... OA36.07 Yang, Yi...... PS01.40, OA30.02 Zhou, Yongzhi...... OA29.03 Yang, Yimin...... OA30.02 Zhu, Guan...... OA46.04 Yassin, Safaa...... OA37.01 Zhu, Xiaojing...... OA17.05 Yates, Elspeth...... PS01.17 Zhu, Xing-Quan.OA04.06, OA35.06, OA45.04, Yates, John...... OA08.01, OA08.02 ...... OA45.05 Yeaman, Sam...... OA24.05 Zintl, Annetta...... OA45.01, OA49.06 Yin, Hong...... OA03.05 Zishiri, Oliver...... PS02.35 Yoo, Mi-Sun...... PS01.47, PS02.38, PS03.44 Zohdy, Sarah...... OA06.03, PS02.19, PS03.20 Yoon, Stephen...... PS02.03 Zuluaga, Daniel...... OA26.05 Youn, Hee-jeong...... PS03.52 Zúñiga-Olivos, Estefanía...... PS01.28 Young, David...... OA44.02 Zynda, Haley...... PS02.25, OA33.02 Yuan, Wang...... PL02.02 Yumi Makatu, Miriam...... PS01.38 Yurayart, Nichapat...... PS01.14

Z

Zacarias Machado, Rosangela...... OA29.01 Zafar, Talha...... OA49.05 Zaid, Taher...... OA49.06 Zajac, Anne...... PS02.69, PS03.23, OA28.02, ...... OA50.02 Zaman, Muhammad Arfan...... PS01.85, ...... PS02.56, PS03.71, OA21.07, OA28.04, ...... OA32.05, OA50.07 Zamanian, Mostafa..PL02.01, PL02.02, PS01.18, ...... PS03.21, PS03.25, PS03.35 Zannou, Olivier...... PS03.47 Zarlenga, Dante....OA07.05, OA12.05, OA24.01, ...... OA36.03 Zawadzki-Baggio, Selma...... OA50.04 Zawistowska-Deniziak, Anna...... PS02.66 Zegbi, Sara...... PS01.83, PS02.47, PS03.57 Zeldenrust, Elizabeth...... PS01.41 Zembsch, Tela...... OA11.05 Zhang, Fu-Kai...... OA35.06, OA45.05 Zhang, Haili...... OA46.04 Zhang, Houshuang...... OA04.05, OA17.01,

WAAVP2019  27th Conference of the World Association for the Advancements of Veterinary Parasitology 64th American Association of Veterinary Parasitologists Annual Meeting 349 63rd Annual Livestock Insect Workers Conference