DOCSLIB.ORG

Unlocking Archived Ticks to Investigate a Novel Putative Pathogen

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Unlocking Archived Ticks to Investigate a Novel Putative Pathogen UNLOCKING ARCHIVED TICKS TO INVESTIGATE A NOVEL PUTATIVE PATHOGEN Anna-Sheree Krige A. S. KRIGE This thesis is presented for the degree of Honours in Molecular Biology at Murdoch University 2017 Declaration I declare that this thesis is my own account of my research and contains as its main content, work that has not been previously submitted for a degree at any tertiary education institution. ……………………………………………………….. Anna-Sheree Krige i Abstract Ticks, as vectors of disease-causing bacteria, represent a significant threat to human and animal health. Of particular concern is the genus Borrelia, which contains several recognised tick-borne pathogens of global health importance. In 2015, the microbiomes within modern Australian ticks were profiled using next- generation sequencing (NGS). Complex communities of bacterial species were revealed, including a novel Borrelia sp. that was identified within a tick parasitising an Australian echidna, Tachyglossus aculeatus. This bacterium, named ‘Candidatus Borrelia tachyglossi’, after its association with the echidna tick, Bothriocroton concolor, forms the fourth known Borrelia group. Following this first report of Borrelia in native Australian ticks, additional questions have emerged concerning whether this bacterium resides in other echidna-biting ticks, and if it has a historic presence in Australia. Around 1,725 tick specimens from 89 registered echidna hosts were collected between 1928 and 2013. Of these, 850 ticks were selected for examination in this study. A total of eight species from three tick genera were morphologically identified. The most common species was B. concolor (89.2%). Interestingly, Amblyomma fimbriatum (22, 2.6%) and Amblyomma triguttatum (19, 2.2%) ticks were recorded for the first time from echidnas. A NGS survey of the bacterial communities within 66 echidna ticks, targeting the V4 region of the 16S rRNA gene, revealed a diverse range of preserved bacteria. Genera of interest included Borrelia (6, 9.1%), Coxiella (19, 28.8%), Ehrlichia (2, 3.0%), Francisella (5, 7.6%), and Rickettsia (23, 34.8%). While NGS of archived echidna tissue biopsies were negative for Borrelia, genus-specific PCR assays amplified the flaB (378 bp) locus in two skin biopsies (2/34; 5.9%) and nine ticks (9/160; 5.6%), with a 99.5-100% similarity to ‘Candidatus B. tachyglossi’ genotypes B and C. Of these positive samples, six ticks and both skin biopsies (6/9, 66.7%; 2/2, 100%) amplified the longer 16S rRNA (1,087 bp) locus, with a 98.2-99.6% similarity to ‘Candidatus B. tachyglossi’ genotype B. Phylogenetic positioning was consistent with previous reports, suggesting ‘Candidatus B. tachyglossi’ formed the fourth Borrelia clade. ii This is the first morphological audit of echidna ticks and NGS survey of the archived tick microbiome. This study has confirmed ‘Candidatus B. tachyglossi’ in Australian echidna ticks in the recent past and for the first time, revealed Borrelia in echidnas. However, whether echidnas are a reservoir for this bacterium could not be established. Additional tick transmission analyses and the examination of echidna blood samples are necessary for confirmation. iii Acknowledgements I would first like to thank my supervisor, Dr. Charlotte Oskam, for your support, advice, and guidance over the course of this research project. There are many people whom I am grateful to for their help and encourgagement. To former research assistant, Annachiara Codello, for ‘showing me the ropes’ in the lab and your kindness. To Kimberly Loh, for introducing me to tick identification and pointing out that ‘those are not eyes’. Also, for your help with Geneious software and providing feedback on one of my earliest drafts, I thank you. To Alexander Gofton, your knowledge and assistance with my library preparation and NGS run was invaluable. Thank you for your time and perseverance with the challenges experienced using this technology. A further thank you goes to the rest of the team at the Vector and Water-Borne Pathogen Research Group for your kind support and advice throughout the year. And to Murdoch University, thank you for granting me a Murdoch University Academic Excellence Award. Finally, I would like to thank my family for their unwavering support and encouragement throughout my honours year. No matter what direction I have decided to pursue, Mum and Dad have always been there to reassure and inspire. And to my two adoring fans and most precious companions, Toby and Lulu, I am so thankful for your company during the many long hours in front of the computer typing up my thesis. Your presence has always given me comfort and to you I dedicate my work. iv Table of Contents Declaration…………………………………………………………………………………………………………i Abstract…………………………………………………………………………………………………………….ii Acknowledgements…………………………………………………………………………………………...iv Table of Contents……………………………………………………………………………………………….v List of Figures……………………………………………………………………………………………………ix List of Tables…………………………………………………………………………………………………….xi List of Abbreviations………………………………………………………………………………………...xii CHAPTER ONE: INTRODUCTION………………………………………………………………………1 1. Introduction………………………………………………………………………………………………....2 1.1 Ticks……………………………………………………………………………………………………...4 1.1.1 Tick morphology……………………………………………………………………………..5 1.1.2 Tick life cycle…….…………………………………………………………………………….7 1.1.3 Ticks in Australia…………………………………………………………………………….8 1.1.4 Tick hosts………………………………………………………………………………………..9 1.1.4.1 Echidnas and echidna-biting ticks…………………………………………9 1.2 Tick-borne pathogens and the tick microbiome……………………………………..10 1.2.1 Tick-borne diseases (TBDs) – a global perspective …………………………12 1.2.2 TBDs in Australia…………………………………………………………………………..13 1.2.3 The genus Borrelia………………………………………………………………………...14 1.2.4 Borrelia in Australia………………………………………………………………………16 1.2.4.1 Borrelia in Australian animals……………………………………………..17 1.2.4.2 Borrelia in Australian ticks………………………………………………….19 1.3 Detection methods of tick-borne pathogens and the tick microbiome………………………………………………………………………………………….20 1.3.1 Polymerase Chain Reaction (PCR)………………………………………………….21 1.3.2 Sanger and next-generation sequencing (NGS) – revolutionary methods of DNA detection……………………………………..22 1.4. Museum specimens – an unexplored ‘gold mine’…………………………………...24 1.4.1 Ancient DNA (aDNA)……………………………………………………………………..25 1.4.1.1 aDNA and NGS – a ‘perfect’ pair…………………………………………..27 1.4.2 Using museum specimens to investigate pathogens………………………..28 v 1.4.2.1 Using museum specimens to detect tick-borne pathogens…………………………………………………………………………..28 1.4.3 Non-destructive sampling – a method for historical preservation………………………………………………………………………………….30 1.5. Research Aims and Hypotheses………………………………...……………………........31 CHAPTER TWO: MATERIALS AND METHODS………………………………………………...33 2. Overview…………………………...………………………………………………...………………………34 2.1 Tick collection and preparation…………………………...………………………………..35 2.1.1 Archived ticks with correlating host tissue…………………………...………...36 2.2 Tick identification…………………………...……………………………………………………36 2.3 Sample collection…………………………...…………………………………………………….37 2.4 DNA extraction…………………………...………………………………………………………..37 2.4.1 Archived echidna ticks…………………………...……………………………………...38 2.4.2 Archived echidna tissue…………………………...…………………………………….39 2.5 Library preparation and NGS…………………………...…………………………………...39 2.6 NGS analysis…………………………...……………………………………………….…………...41 2.7 PCR assays…………………………...………………………………………………………………42 2.7.1 16S Mam qPCR assay of echidna tissue…………………………...……………...44 2.7.1.1 16S Mam conventional PCR assay of correlating echidna tissue ticks…………………………...…………………………….….44 2.7.2 Borrelia-specific PCR assays of echidna tissue and ticks…………………..45 2.7.2.1 flaB nested-PCR assay…………………………...……………………………45 2.7.2.2 Borrelia-specific 16S nested-PCR assay…………………………...…..46 2.8 Gel electrophoresis…………………………...………………………………………………….47 2.9 PCR purification…………………………...………………………………………………………47 2.10 Sanger sequencing and BLAST…………………………...……………………………….48 2.11 Phylogenetic analysis…………………………...………………………………………….…49 2.12 Percentage of Borrelia-infected echidna ticks………………………………………50 CHAPTER THREE: RESULTS……………………………………………………………………………51 3. Results…………………………………………………………………………………………………………52 3.1 Morphological identification of archived echidna-biting ticks………………...52 3.1.1 Sample selection for DNA extraction and analyses…………………………..54 vi 3.2 NGS………….…………………………………………………………………………………………..56 3.3 PCR assays………..………………………………………………………………………………….62 3.3.1 16S Mam qPCR assay of echidna tissue………………………..……..…………..62 3.3.1.1 16S Mam PCR assay of correlating tissue ticks…………………..…64 3.3.2 flaB nested-PCR assay……………………………………….……………….…………..66 3.3.2.1 The flaB gene in echidna tissue…………………………………………...68 3.3.2.2 The flaB gene in echidna ticks……………………………………………..70 3.3.3 Borrelia-specific 16S nested-PCR assay………………………………………..…72 3.4 Sanger sequencing and BLAST…………………………………..………………………….72 3.5 Phylogenetic analysis………………………..………………………………………………….73 3.6 Percentage of Borrelia-infected echidna ticks….………………………………….….76 CHAPTER FOUR: DISCUSSION AND CONCLUSION…………………………………………..79 4. Discussion……………………………………………………………………………………………………80 4.1 Morphological identification of archived echidna-biting ticks………………...80 4.2 NGS bacterial profiling of archived echidna-biting ticks…………………………82 4.3 Detecting a novel Borrelia species in archived echidna-biting ticks………...85 4.4 Future directions………………………………………….………………………………………88 4.5 Conclusion………………………………………….………………………………………………..89 REFERENCES…………………………………………………………………………………………………..90 References……………………………………………………………………………………………………....91
Load more

Recommended publications
  • Molecular Evidence of Novel Spotted Fever Group Rickettsia
    pathogens Article Molecular Evidence of Novel Spotted Fever Group Rickettsia Species in Amblyomma albolimbatum Ticks from the Shingleback Skink (Tiliqua rugosa) in Southern Western Australia Mythili Tadepalli 1, Gemma Vincent 1, Sze Fui Hii 1, Simon Watharow 2, Stephen Graves 1,3 and John Stenos 1,* 1 Australian Rickettsial Reference Laboratory, University Hospital Geelong, Geelong 3220, Australia; [email protected] (M.T.); [email protected] (G.V.); [email protected] (S.F.H.); [email protected] (S.G.) 2 Reptile Victoria Inc., Melbourne 3035, Australia; [email protected] 3 Department of Microbiology and Infectious Diseases, Nepean Hospital, NSW Health Pathology, Penrith 2747, Australia * Correspondence: [email protected] Abstract: Tick-borne infectious diseases caused by obligate intracellular bacteria of the genus Rick- ettsia are a growing global problem to human and animal health. Surveillance of these pathogens at the wildlife interface is critical to informing public health strategies to limit their impact. In Australia, reptile-associated ticks such as Bothriocroton hydrosauri are the reservoirs for Rickettsia honei, the causative agent of Flinders Island spotted fever. In an effort to gain further insight into the potential for reptile-associated ticks to act as reservoirs for rickettsial infection, Rickettsia-specific PCR screening was performed on 64 Ambylomma albolimbatum ticks taken from shingleback skinks (Tiliqua rugosa) lo- cated in southern Western Australia. PCR screening revealed 92% positivity for rickettsial DNA. PCR Citation: Tadepalli, M.; Vincent, G.; amplification and sequencing of phylogenetically informative rickettsial genes (ompA, ompB, gltA, Hii, S.F.; Watharow, S.; Graves, S.; Stenos, J.
    [Show full text]
  • Genus Boophilus Curtice Genus Rhipicentor Nuttall & Warburton
    3 CONTENTS General remarks 4 Genus Amblyomma Koch 5 Genus Anomalohimalaya Hoogstraal, Kaiser & Mitchell 46 Genus Aponomma Neumann 47 Genus Boophilus Curtice 58 Genus Hyalomma Koch. 63 Genus Margaropus Karsch 82 Genus Palpoboophilus Minning 84 Genus Rhipicentor Nuttall & Warburton 84 Genus Uroboophilus Minning. 84 References 86 SUMMARI A list of species and subspecies currently included in the tick genera Amblyomma, Aponomma, Anomalohimalaya, Boophilus, Hyalomma, Margaropus, and Rhipicentor, as well as in the unaccepted genera Palpoboophilus and Uroboophilus is given in this paper. The published synonymies and authors of each spécifie or subspecific name are also included. Remaining tick genera have been reviewed in part in a previous paper of this series, and will be finished in a future third part. Key-words: Amblyomma, Aponomma, Anomalohimalaya, Boophilus, Hyalomma, Margaropus, Rhipicentor, Uroboophilus, Palpoboophilus, species, synonymies. RESUMEN Se proporciona una lista de las especies y subespecies actualmente incluidas en los géneros Amblyomma, Aponomma, Anomalohimalaya, Boophilus, Hyalomma, Margaropus y Rhipicentor, asi como en los géneros no aceptados Palpoboophilus and Uroboophilus. Se incluyen también las sinonimias publicadas y los autores de cada nombre especifico o subespecifico. Los restantes géneros de garrapatas han sido revisados en parte en un volumen previo de esta serie, y serân terminados en una futura tercera parte. Palabras claves Amblyomma, Aponomma, Anomalohimalaya, Boophilus, Hyalomma, Margaropus, Rhipicentor, Uroboophilus, Palpoboophilus, especies, sinonimias. 4 GENERAL REMARKS Following is a list of species and subspecies of ticks d~e scribed in the genera Amblyomma, Aponomma, Anomalohimalaya, Boophilus, Hyalorma, Margaropus, and Rhipicentor, as well as in the unaccepted genera Palpoboophilus and Uroboophilus. The first volume (Estrada- Pena, 1991) included data for Haemaphysalis, Anocentor, Dermacentor, and Cosmiomma.
    [Show full text]
  • Running Title: Bacterial Community Profiling Highlights Complex Diversity and Novel Organisms in Wildlife Ticks. Authors: Siobho
    bioRxiv preprint doi: https://doi.org/10.1101/807131; this version posted October 17, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Running title: Bacterial community profiling highlights complex diversity and novel 2 organisms in wildlife ticks. 3 4 Authors: Siobhon L. Egan1, Siew-May Loh1, Peter B. Banks2, Amber Gillett3, Liisa A. 5 Ahlstrom4, Una M. Ryan1, Peter J. Irwin1 and Charlotte L. Oskam1,* 6 7 1 Vector and Waterborne Pathogens Research Group, College of Science, Health, 8 Engineering and Education, Murdoch University, Perth, Western Australia, Australia 9 2 School of Life and Environmental Sciences, The University of Sydney, Sydney, New South 10 Wales, Australia 11 3 Australia Zoo Wildlife Hospital, Beerwah, Queensland, Australia 12 4 Bayer Australia Ltd, Animal Health, Pymble, New South Wales, Australia 13 * Corresponding author 14 15 S.L.E [email protected]; https://orcid.org/0000-0003-4395-4069 16 S-M.L. [email protected] 17 P.B.B. [email protected]; https://orcid.org/0000-0002-4340-6495 18 A.G. [email protected] 19 L.A.A [email protected] 20 U.M.R. [email protected]; https://orcid.org/0000-0003-2710-9324 21 P.J.I. [email protected]; https://orcid.org/0000-0002-0006-8262 22 C.L.O. c.o [email protected] ; https://orcid.org/0000-0001-8886-2120 23 24 Abstract 25 Ticks (Acari: Ixodida) transmit a greater variety of pathogens than any other blood-feeding 1 bioRxiv preprint doi: https://doi.org/10.1101/807131; this version posted October 17, 2019.
    [Show full text]
  • Download Download
    HAMADRYAD Vol. 27. No. 2. August, 2003 Date of issue: 31 August, 2003 ISSN 0972-205X CONTENTS T. -M. LEONG,L.L.GRISMER &MUMPUNI. Preliminary checklists of the herpetofauna of the Anambas and Natuna Islands (South China Sea) ..................................................165–174 T.-M. LEONG & C-F. LIM. The tadpole of Rana miopus Boulenger, 1918 from Peninsular Malaysia ...............175–178 N. D. RATHNAYAKE,N.D.HERATH,K.K.HEWAMATHES &S.JAYALATH. The thermal behaviour, diurnal activity pattern and body temperature of Varanus salvator in central Sri Lanka .........................179–184 B. TRIPATHY,B.PANDAV &R.C.PANIGRAHY. Hatching success and orientation in Lepidochelys olivacea (Eschscholtz, 1829) at Rushikulya Rookery, Orissa, India ......................................185–192 L. QUYET &T.ZIEGLER. First record of the Chinese crocodile lizard from outside of China: report on a population of Shinisaurus crocodilurus Ahl, 1930 from north-eastern Vietnam ..................193–199 O. S. G. PAUWELS,V.MAMONEKENE,P.DUMONT,W.R.BRANCH,M.BURGER &S.LAVOUÉ. Diet records for Crocodylus cataphractus (Reptilia: Crocodylidae) at Lake Divangui, Ogooué-Maritime Province, south-western Gabon......................................................200–204 A. M. BAUER. On the status of the name Oligodon taeniolatus (Jerdon, 1853) and its long-ignored senior synonym and secondary homonym, Oligodon taeniolatus (Daudin, 1803) ........................205–213 W. P. MCCORD,O.S.G.PAUWELS,R.BOUR,F.CHÉROT,J.IVERSON,P.C.H.PRITCHARD,K.THIRAKHUPT, W. KITIMASAK &T.BUNDHITWONGRUT. Chitra burmanica sensu Jaruthanin, 2002 (Testudines: Trionychidae): an unavailable name ............................................................214–216 V. GIRI,A.M.BAUER &N.CHATURVEDI. Notes on the distribution, natural history and variation of Hemidactylus giganteus Stoliczka, 1871 ................................................217–221 V. WALLACH.
    [Show full text]
  • Conservation of South African Tortoises with Emphasis on Their Apicomplexan Haematozoans, As Well As Biological and Metal-Fingerprinting of Captive Individuals
    CONSERVATION OF SOUTH AFRICAN TORTOISES WITH EMPHASIS ON THEIR APICOMPLEXAN HAEMATOZOANS, AS WELL AS BIOLOGICAL AND METAL-FINGERPRINTING OF CAPTIVE INDIVIDUALS By Courtney Antonia Cook THESIS submitted in fulfilment of the requirements for the degree PHILOSOPHIAE DOCTOR (Ph.D.) in ZOOLOGY in the FACULTY OF SCIENCE at the UNIVERSITY OF JOHANNESBURG Supervisor: Prof. N. J. Smit Co-supervisors: Prof. A. J. Davies and Prof. V. Wepener June 2012 “We need another and a wiser and perhaps a more mystical concept of animals. Remote from universal nature, and living by complicated artifice, man in civilization surveys the creature through the glass of his knowledge and sees thereby a feather magnified and the whole image in distortion. We patronize them for their incompleteness, for their tragic fate of having taken form so far below ourselves. And therein we err, and greatly err. For the animal shall not be measured by man. In a world older and more complete than ours they move finished and complete, gifted with extensions of the senses we have lost or never attained, living by voices we shall never hear. They are not brethren, they are not underlings; they are other nations caught with ourselves in the net of life and time, fellow prisoners of the splendour and travail of the earth.” Henry Beston (1928) ABSTRACT South Africa has the highest biodiversity of tortoises in the world with possibly an equivalent diversity of apicomplexan haematozoans, which to date have not been adequately researched. Prior to this study, five apicomplexans had been recorded infecting southern African tortoises, including two haemogregarines, Haemogregarina fitzsimonsi and Haemogregarina parvula, and three haemoproteids, Haemoproteus testudinalis, Haemoproteus balazuci and Haemoproteus sp.
    [Show full text]
  • Rickettsia Detected in the Reptile Tick Bothriocroton Hydrosauri from the Lizard Tiliqua Rugosa in South Australia
    pathogens Article Rickettsia Detected in the Reptile Tick Bothriocroton hydrosauri from the Lizard Tiliqua rugosa in South Australia Harriet Whiley 1,†, Georgie Custance 2,†, Stephen Graves 3, John Stenos 3, Michael Taylor 1, Kirstin Ross 1 and Michael G. Gardner 2,4,* 1 School of the Environment, Health and the Environment, Flinders University, GPO BOX 2100, Adelaide 5001, Australia; harriet.whiley@flinders.edu.au (H.W.); michael.taylor@flinders.edu.au (M.T.); [email protected] (K.R.) 2 School of Biological Sciences, Flinders University, GPO Box 2100, Adelaide 5001, Australia; cust0014@flinders.edu.au 3 Australian Rickettsial Reference Laboratory, Barwon Health, Geelong Hospital, PO BOX 281, Geelong 3220, Australia; [email protected] (S.G.); [email protected] (J.S.) 4 Evolutionary Biology Unit, South Australian Museum, North Terrace, Adelaide 5000, Australia * Correspondence: michael.gardner@flinders.edu.au; Tel.: +61-8-201-2315; Fax: +61-8-201-3015 † These authors contributed equally to this work. Academic Editor: Lawrence S. Young Received: 27 January 2016; Accepted: 7 June 2016; Published: 8 June 2016 Abstract: Rickettsiosis is a potentially fatal tick borne disease. It is caused by the obligate intracellular bacteria Rickettsia, which is transferred to humans through salivary excretions of ticks during the biting process. Globally, the incidence of tick-borne diseases is increasing; as such, there is a need for a greater understanding of tick–host interactions to create more informed risk management strategies. Flinders Island spotted fever rickettsioses has been identified throughout Australia (Tasmania, South Australia, Queensland and Torres Strait Islands) with possible identifications in Thailand, Sri Lanka and Italy.
    [Show full text]
  • Approaches on Arthropod´S Molecular Systematics Studies
    Int.J.Curr.Microbiol.App.Sci (2014) 3(10) 156-168 ISSN: 2319-7706 Volume 3 Number 10 (2014) pp. 156-168 http://www.ijcmas.com Review Article Approaches on Arthropod´s Molecular Systematics Studies José Brites-Neto1,3, Gino Chaves da Rocha2, Luiz Humberto Gomes3 and Keila Maria Roncato Duarte4 1Epidemiological Surveillance Department, Secretary of Health, Americana, São Paulo, Brazil 2Faculty of Agronomy and Veterinary, University of Brasília, Brazil 3Department of Exact Sciences/ESALQ/USP, São Paulo, Brazil; 4Institute of Animal Science of Nova Odessa, São Paulo, Brazil *Corresponding author A B S T R A C T Phylogenies moleculars provide means for analyzing many old problems of behavioral and eco-epidemiology of arthropods, many of which are importance for public health. A number of molecular markers is used in studies of phylogeny, K e y w o r d s ecology and population dynamics. Mitochondrial DNA is the most widely used Ribosomal marker of DNA regions for insects as well as for animals in general. Nuclear DNA, ribosomal RNA genes have been widely used, especially due to their abundance in mitochondrial the genome and its ease of amplification and sequencing of highly conserved and DNA, variable regions (containing the 18S, 5.8S and 28S genes separated by non polymorphis transcribed and transcribed spacers). Questions about the intraspecific population m, molecular structure, species limits, and the diagnosis of species, can be often elucidated by markers, techniques such as allozyme electrophoresis, restriction fragment length phylogenies. polymorphism (RFLP), or any variety of new techniques for DNA-based, as single strand conformation polymorphism (SSCP), amplified fragment length polymorphism DNA (AFLP) or random amplified polymorphic DNA (RAPD).
    [Show full text]
  • Molecular Barcoding of Australian Ticks
    Molecular barcoding of Australian ticks by Megan Evans Bachelor of Science This thesis is presented for the degree of Bachelor of Science Honours in Biological Sciences School of Veterinary and Life Sciences Murdoch University, 2018 1 Author’s Declaration I declare that this thesis is my own account of my research and contains as its main content work which has not previously been submitted for a degree at any tertiary education institution. Megan Evans ii Abstract Globally, ticks (Acari: Ixodida) are one of the most important vectors of disease due to their ability to transmit a wide variety of pathogenic bacteria, protozoa, and viruses during blood feeding. The microbes transmitted by ticks varies by species, and so it is essential that ticks are able to be identified correctly. However, the identification and discrimination of tick species still relies on traditional morphological techniques, which can at times be ambiguous, particularly in the case of subadult ticks. This study tested previously developed molecular barcoding assays to facilitate the identification of Australian ticks using DNA sequences in the case that morphology is inconclusive. Using reference ticks from eight native species of medical and veterinary importance (Amblyomma triguttatum, Bothriocroton auruginans, Haemaphysalis bancrofti, Haemaphysalis humerosa, Ixodes cornuatus, Ixodes hirsti, Ixodes holocyclus and Ixodes tasmani), four potential barcoding genes were trialled (Cytochrome c oxidase (COI), Internal transcribed spacer 2 (ITS2), 16S rRNA and 12S rRNA). Amplification was successful in 98.3% of samples (n=58) for COI, 89.6% (n=48) of samples for ITS2, and 100% of samples for both 16S rRNA and 12S rRNA (n=58 and n=48 respectively).
    [Show full text]
  • Checklist of the Mites of Pakistan
    Zootaxa 4464 (1): 001–178 ISSN 1175-5326 (print edition) http://www.mapress.com/j/zt/ Monograph ZOOTAXA Copyright © 2018 Magnolia Press ISSN 1175-5334 (online edition) https://doi.org/10.11646/zootaxa.4464.1.1 http://zoobank.org/urn:lsid:zoobank.org:pub:182B2DAF-5C88-4407-8B07-DD06F23E5854 ZOOTAXA 4464 Checklist of the mites of Pakistan BRUCE HALLIDAY1, MUHAMMAD KAMRAN2 & MUHAMMAD HAMID BASHIR3 1 Australian National Insect Collection, CSIRO, GPO Box 1700, Canberra ACT 2601, Australia. E-mail: [email protected] 2 Department of Plant Protection, College of Food and Agriculture Sciences, King Saud University, P.O. Box 2460, Al-Riyadh, 11451, Saudi Arabia. E-mail: [email protected], [email protected] 3 Department of Agricultural Entomology, University of Agriculture, Faisalabad, Pakistan. E-mail: [email protected] Magnolia Press Auckland, New Zealand Accepted by O. Seeman: 10 Jul. 2018; published: 30 Aug. 2018 Licensed under a Creative Commons Attribution License http://creativecommons.org/licenses/by/3.0 BRUCE HALLIDAY, MUHAMMAD KAMRAN & MUHAMMAD HAMID BASHIR Checklist of the mites of Pakistan (Zootaxa 4464) 178 pp.; 30 cm. 30 Aug. 2018 ISBN 978-1-77670-438-5 (paperback) ISBN 978-1-77670-439-2 (Online edition) FIRST PUBLISHED IN 2018 BY Magnolia Press P.O. Box 41-383 Auckland 1346 New Zealand e-mail: [email protected] http://www.mapress.com/j/zt © 2018 Magnolia Press ISSN 1175-5326 (Print edition) ISSN 1175-5334 (Online edition) 2 · Zootaxa 4464 (1) © 2018 Magnolia Press HALLIDAY Table of contents Abstract . 3 Introduction . 3 Methods . 4 Astigmata. 5 Cryptostigmata. 19 Ixodida . 38 Mesostigmata .
    [Show full text]
  • Tick Mitochondrial Genomes: Structural Characteristics and Phylogenetic Implications Tianhong Wang, Shiqi Zhang, Tingwei Pei, Zhijun Yu* and Jingze Liu*
    Wang et al. Parasites Vectors (2019) 12:451 https://doi.org/10.1186/s13071-019-3705-3 Parasites & Vectors REVIEW Open Access Tick mitochondrial genomes: structural characteristics and phylogenetic implications Tianhong Wang, Shiqi Zhang, Tingwei Pei, Zhijun Yu* and Jingze Liu* Abstract Ticks are obligate blood-sucking arachnid ectoparasites from the order Acarina, and many are notorious as vectors of a wide variety of zoonotic pathogens. However, the systematics of ticks in several genera is still controversial. The mitochondrial genome (mt-genome) has been widely used in arthropod phylogeny, molecular evolution and popula- tion genetics. With the development of sequencing technologies, an increasing number of tick mt-genomes have been sequenced and annotated. To date, 63 complete tick mt-genomes are available in the NCBI database, and these genomes have become an increasingly important genetic resource and source of molecular markers in phylogenetic studies of ticks in recent years. The present review summarizes all available complete mt-genomes of ticks in the NCBI database and analyses their characteristics, including structure, base composition and gene arrangement. Further- more, a phylogenetic tree was constructed using mitochondrial protein-coding genes (PCGs) and ribosomal RNA (rRNA) genes from ticks. The results will provide important clues for deciphering new tick mt-genomes and establish a foundation for subsequent taxonomic research. Keywords: Ticks, Mitochondrial genome (mt-genome), Gene structure, Phylogeny Background fnancial losses due to ticks and TBDs are in the billions Ticks are obligate blood-sucking arachnid ectoparasites of dollars [3, 11]. A total of 896 tick species have been that can feed on a wide range of vertebrates, including described worldwide in three families: Ixodidae (hard mammals, birds and reptiles [1, 2].
    [Show full text]
  • Bacterial Community Profiling of Western Australian Bobtail Lizard (Tiliqua Rugosa) Ticks
    BACTERIAL COMMUNITY PROFILING OF WESTERN AUSTRALIAN BOBTAIL LIZARD (TILIQUA RUGOSA) TICKS Ruby Mckenna Bachelor of Science Submitted in fulfilment of the requirements for the degree of Honours in Biological Sciences School of Veterinary and Life Sciences Murdoch University, Perth 2019 iii Statement of Original Authorship I declare that this thesis is my own account of my research and contains as its main content work which has not previously been submitted for a degree at any tertiary education institution. Ruby McKenna iv Abstract Ticks are haematophagous arthropods and major vectors of pathogenic microorganisms. In Australia, over 74 species of ticks have been described, of which 13 are known to parasitise reptiles. While only three tick borne diseases are formally recognised, Rickettsia honei, the causative agent of Flinders Island spotted fever, has been long associated with the reptile tick, Bothriocroton hydrosauri, despite this tick rarely reported to parasitise people. More recently, a novel Rickettsia species, Rickettsia gravesii, was reported in the ornate kangaroo tick (a common human biting tick), Amblyomma triguttatum, on Barrow Island in Western Australia. In addition, a number of overseas tick-associated microbes (taxa of interest) have been identified in Australian human-biting ticks using an advanced molecular technique, next generation sequencing (NGS). Therefore, the aims of this study were to morphologically and molecularly identify ticks that parasitise reptiles, specifically the bobtail lizard, Tiliqua rugosa, and to employ NGS to profile the bacterial 16S rRNA gene (16S) within the ticks. The taxa identified would then be phylogenetically compared to known taxa of interest. A total of 306 ticks from Western Australia were morphologically identified and 30 were removed from the data set and the remaining 276 included all developmental stages comprising males 43.1% (n=119); nymphs 40.6% (n=112); females 15.2% (n=42) and three larvae (1.1%).
    [Show full text]
  • Insecticidal Proteins and Methods for Their Use Insektizidproteine Und Verfahren Zu Deren Verwendung Protéines Insecticides Et Leurs Procédés D’Utilisation
    (19) *EP003102684B1* (11) EP 3 102 684 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12N 15/82 (2006.01) C07K 14/415 (2006.01) 06.05.2020 Bulletin 2020/19 (86) International application number: (21) Application number: 15708059.9 PCT/US2015/014816 (22) Date of filing: 06.02.2015 (87) International publication number: WO 2015/120270 (13.08.2015 Gazette 2015/32) (54) INSECTICIDAL PROTEINS AND METHODS FOR THEIR USE INSEKTIZIDPROTEINE UND VERFAHREN ZU DEREN VERWENDUNG PROTÉINES INSECTICIDES ET LEURS PROCÉDÉS D’UTILISATION (84) Designated Contracting States: (74) Representative: J A Kemp LLP AL AT BE BG CH CY CZ DE DK EE ES FI FR GB 14 South Square GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO Gray’s Inn PL PT RO RS SE SI SK SM TR London WC1R 5JJ (GB) (30) Priority: 07.02.2014 US 201461937288 P (56) References cited: WO-A2-2013/098858 (43) Date of publication of application: 14.12.2016 Bulletin 2016/50 • H. K. ABICHT ET AL: "Genome Sequence of Desulfosporosinus sp. OT, an Acidophilic (73) Proprietors: Sulfate-Reducing Bacterium from Copper Mining • Pioneer Hi-Bred International, Inc. Waste in Norilsk, Northern Siberia", JOURNAL Johnston, Iowa 50131-1014 (US) OF BACTERIOLOGY, vol. 193, no. 21, 1 November • E. I. du Pont de Nemours and Company 2011 (2011-11-01), pages 6104-6105, Wilmington, DE 19805 (US) XP055196331, ISSN: 0021-9193, DOI: 10.1128/JB.06018-11 -& DATABASE UniProt (72) Inventors: [Online] 16 November 2011 (2011-11-16), • BARRY, Jennifer "SubName: Full=Uncharacterized protein Ames, Iowa 50014 (US) {ECO:0000313|EMBL:EGW36042.1};", • HAYES, Kevin XP002741068, retrieved from EBI accession no.
    [Show full text]