DOCSLIB.ORG

Characterization of Secreted Giardia Intestinalis Cysteine Proteases

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterization of Secreted Giardia Intestinalis Cysteine Proteases Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 1763 Characterization of secreted Giardia intestinalis cysteine proteases JINGYI LIU ACTA UNIVERSITATIS UPSALIENSIS ISSN 1651-6214 ISBN 978-91-513-0551-6 UPPSALA urn:nbn:se:uu:diva-372997 2019 Dissertation presented at Uppsala University to be publicly examined in A1:111a, Uppsala Biomedicinska Centrum BMC, Husarg. 3, Uppsala, Thursday, 28 February 2019 at 09:15 for the degree of Doctor of Philosophy. The examination will be conducted in English. Faculty examiner: Professor Andre Buret (University of Calgary). Abstract Liu, J. 2019. Characterization of secreted Giardia intestinalis cysteine proteases. Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 1763. 72 pp. Uppsala: Acta Universitatis Upsaliensis. ISBN 978-91-513-0551-6. Giardia intestinalis, the causative agent of the diarrheal disease giardiasis, is a protozoan parasite that colonizes the upper small intestine of mammals, including humans. It can be divided into eight genotypes or assemblages (A through H) and only assemblage A and B are infective to humans. Giardiasis is a multi-factorial disease but few giardial virulence factors have been identified and characterized. In this thesis, we used proteomics to identify the major excretory-secretory products (ESPs) released by Giardia trophozoites of the WB and GS isolates during interaction with intestinal epithelial cells (IECs) in vitro (Paper I). To deepen our understanding of the role of ESPs in giardiasis, we focused on three specific secreted Giardia cysteine proteases (CPs; CP14019, CP16160 and CP16779). All the three CPs are capable of opening the apical junction complexes between IECs to degrade chemokines produced in response to Giardia (Paper II). This can partly explain the induction of symptoms and immunosuppression seen during giardiasis. We further studied the cleavage specificity of these CPs using substrate phage display and recombinant protein substrates. The preferred sequences were used to search potential human in vivo targets and a number of candidates were identified, including human immunoglobulins as well as defensins, that were subsequently shown to be efficiently cleaved by the CPs (Paper III). To investigate the involvement of CPs in mucus degradation, we tested the CPs on recombinant MUC2 constructs and full-length MUC2. MUC2 is the major component of the mucus layer in the small intestine. It was shown that CP14019 cleave MUC2 in the N-terminal, suggesting a mechanism that the parasite can use to disrupt/release the mucus gel network and get access to the intestinal epithelium of the host (Paper IV). In summary, this thesis has studied secreted Giardia CPs and their roles in Giardia infections, providing significant insights into the molecular pathogenesis of giardiasis. Keywords: Giardia intestinalis, ESPs, CPs, apical junction, chemokines, phage display, immunoglobulins, defensins, mucin. Jingyi Liu, Department of Cell and Molecular Biology, Microbiology, Box 596, Uppsala University, SE-75124 Uppsala, Sweden. © Jingyi Liu 2019 ISSN 1651-6214 ISBN 978-91-513-0551-6 urn:nbn:se:uu:diva-372997 (http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-372997) To my family List of Papers This thesis is based on the following papers, which are referred to in the text by their Roman numerals. I Ma'ayeh, S.Y., Liu, J., Peirasmaki, D., Hornaeus, K., Bergström Lind, S., Grabherr, M., Bergquist, J., and Svärd, S.G. (2017). Characterization of the Giardia intestinalis secretome during interaction with human intestinal epithelial cells: The impact on host cells. PLoS Negl Trop Dis, 11(12): e0006120. II Liu, J., Ma'ayeh, S., Peirasmaki, D., Lundström-Stadelmann, B., Hellman, L., and Svärd, S.G. (2018). Secreted Giardia intestinalis cysteine proteases disrupt intestinal epithelial cell junctional complexes and degrade chemokines. Virulence, 9(1): 879-894. III Liu, J., Fu, Z., Hellman, L., and Svärd, S.G (2018). Cleavage specificity of recombinant Giardia intestinalis cysteine proteas- es: Degradation of immunoglobulins and defensins. Mol Bio- chem Parasitol, 227: 29-38. IV Arike, L.*, Recktenwald, C.V.*, Liu, J., Trillo-Muyo, S., Svärd, S.G., Hansson, G.C (2019). MUC2 mucin is cleaved by Giardia intestinalis cysteine protease 14019. Manuscript *These authors contributed equally Reprints were made with permission from the respective publishers. Contents Introduction ................................................................................................... 11 Giardia ..................................................................................................... 11 Life cycle of Giardia ................................................................................ 12 The trophozoite .................................................................................... 13 Cell differentiation ............................................................................... 16 Giardiasis .................................................................................................. 17 Giardia pathogenesis ................................................................................ 17 Microvilli shortening, destruction, malabsorption ............................... 19 Intestinal barrier disruption .................................................................. 19 Intestinal microbiota ............................................................................ 21 Mucus layer ......................................................................................... 23 Host cytokine profile ........................................................................... 26 Virulence factors ...................................................................................... 27 Attachment ........................................................................................... 28 Antigenic variation .............................................................................. 29 ADI and OCT ...................................................................................... 30 Cysteine proteases ............................................................................... 31 Other factors ........................................................................................ 33 Current investigation ..................................................................................... 35 Secretome study of IECs-parasites interplay (Paper I) ............................. 35 Secreted Giardia CPs are key virulence factors (Papers II, III and IV) ... 37 Tight junction disruption and chemokine degradation (Paper II) ........ 37 Degradation of immunoglobulins and defensins (Paper III) ................ 40 Mucin degradation (Paper IV) ............................................................. 41 Conclusion and future perspectives .............................................................. 43 Cleavage specificity of other secreted Giardia CPs ................................. 43 Knockout study of Giardia CPs ............................................................... 44 CPs role in vaccines and as drug targets .................................................. 45 How are the activities of CPs regulated? .................................................. 45 Assemblage-specific differences of CPs .................................................. 46 Svensk sammanfattning ................................................................................ 47 Acknowledgements ....................................................................................... 49 References ..................................................................................................... 51 Abbreviations ADI Arginine deiminase AJ Adherence junction AME Arginine metabolizing enzyme AMP Antimicrobial peptide AP Adaptor protein ATP Adenosine triphosphate CCL (-2, -20) Chemokine with double cysteine (CC) N-terminal motif COP Coat protein CXCL (-1, -2, -3) Chemokine with CC motif separated by 1 amino acid CP Cysteine protease DC Dendritic cell EF-1α Elongation factor 1 alpha ER Endoplasmic reticulum ESP Excretory-secretory product FeS Iron-sulfur HCMP High-cysteine membrane protein IBS Irritable bowel syndrome IEC Intestinal epithelial cell Ig (-A, -G) Immunoglobulin A, G LC-MS Liquid chromatography-mass spectrometry MLCK Myosin light chain kinase MMP Matrix metalloprotease 7 NO Nitric oxide NOS Nitric oxide synthase OCT Ornithine carbamoyl transferase PV Peripheral vacuole SDS-PAGE Sodium dodecyl sulphate-polyacrylamide gel electro- phoresis TJ Tight junction VSP Variant-specific surface protein ZO Zonula occludens α-HD (-5,-6) α-defensin (-5,-6) β-HD1 β-defensin 1 Introduction Our understanding of Giardia’s pathogenesis is increasing, partially attribut- ed to the understanding of the effects of excretory-secretory products (ESPs) released by Giardia during host-parasite interactions. However, the under- standing is limited to effects induced by culture supernatants in most studies and the specific virulence factors have rarely been identified and character- ized. Proteases constitute a large proportion of the ESPs and secreted prote- ases have been suggested to be key players in the pathogenesis of many pro- tozoan parasitic diseases, including giardiasis. Giardia Giardia was initially described by van Leeuwenhoek in 1681 when he was examining his own diarrheal stools under his primitive microscope (Dobell, 1920). The organism was described in more detail by Lambl
Load more

Recommended publications
  • Five Facts About Giardia Lamblia
    PEARLS Five facts about Giardia lamblia Lenka Cernikova, Carmen FasoID, Adrian B. HehlID* Laboratory of Molecular Parasitology, Institute of Parasitology, University of Zurich (ZH), Zurich, Switzerland * [email protected] Fact 1: Infection with Giardia lamblia is one of the most common causes of waterborne nonbacterial and nonviral diarrheal disease G. lamblia (syn. intestinalis, duodenalis) is a zoonotic enteroparasite. It proliferates in an extra- cellular and noninvasive fashion in the small intestine of vertebrate hosts, causing the diarrheal disease known as giardiasis. Virtually all mammals can be infected with G. lamblia, and epide- miological data point to giardiasis as a zoonosis [1]. Infections in humans may be asymptom- atic or associated with diarrhea, malabsorption, bloating, abdominal pain, fatigue, and weight loss. Based on the latest figures provided by WHO, G. lamblia is the third most common agent of diarrheal disease worldwide with over 300 million reported cases per annum, preceded only a1111111111 by rotavirus and Cryptosporidium parvum and hominis in the most vulnerable target group of a1111111111 a1111111111 children under five years of age [2]. The prevalence of giardiasis in humans ranges from 2%± a1111111111 3% in industrialized countries, up to 30% in low-income and developing countries [3]. Giardi- a1111111111 asis was formerly included in the WHO neglected diseases initiative and is directly associated with poverty and poor quality of drinking water [4]. Acute infection develops over a period of three weeks, peaking at eight days post infection. Generally, healthy hosts clear the infection within 2±3 weeks, whereas the occasional chronically infected host shows signs of villus and crypt atrophy, enterocyte apoptosis, and ultimately severe disruption of epithelial barrier func- OPEN ACCESS tion [5].
    [Show full text]
  • A Multifaceted Approach to Combating Leishmaniasis, a Neglected Tropical Disease
    OLD TARGETS AND NEW BEGINNINGS: A MULTIFACETED APPROACH TO COMBATING LEISHMANIASIS, A NEGLECTED TROPICAL DISEASE DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy from the Graduate School of The Ohio State University By Adam Joseph Yakovich, B.S. ***** The Ohio State University 2007 Dissertation Committee: Karl A Werbovetz, Ph.D., Advisor Approved by Pui-Kai Li, Ph.D. Werner Tjarks, Ph.D. ___________________ Ching-Shih Chen, Ph.D Advisor Graduate Program In Pharmacy ABSTRACT Leishmaniasis, a broad spectrum of disease which is caused by the protozoan parasite Leishmania , currently affects 12 million people in 88 countries worldwide. There are over 2 million of new cases of leishmaniasis occurring annually. Clinical manifestations of leishmaniasis range from potentially disfiguring cutaneous leishmaniasis to the most severe manifestation, visceral leishmaniasis, which attacks the reticuloendothelial system and has a fatality rate near 100% if left untreated. All currently available therapies all suffer from drawbacks including expense, route of administration and developing resistance. In the laboratory of Dr. Karl Werbovetz our primary goal is the identification and development of an inexpensive, orally available antileishmanial chemotherapeutic agent. Previous efforts in the lab have identified a series of dinitroaniline compounds which have promising in vitro activity in inhibiting the growth of Leishmania parasites. It has since been discovered that these compounds exert their antileishmanial effects by binding to tubulin and inhibiting polymerization. Remarkably, although mammalian and Leishmania tubulins are ~84 % identical, the dinitroaniline compounds show no effect on mammalian tubulin at concentrations greater than 10-fold the IC 50 value determined for inhibiting Leishmania tubulin ii polymerization.
    [Show full text]
  • Giardiasis Importance Giardiasis, a Gastrointestinal Disease Characterized by Acute Or Chronic Diarrhea, Is Caused by Protozoan Parasites in the Genus Giardia
    Giardiasis Importance Giardiasis, a gastrointestinal disease characterized by acute or chronic diarrhea, is caused by protozoan parasites in the genus Giardia. Giardia duodenalis is the major Giardia Enteritis, species found in mammals, and the only species known to cause illness in humans. This Lambliasis, organism is carried in the intestinal tract of many animals and people, with clinical signs Beaver Fever developing in some individuals, but many others remaining asymptomatic. In addition to diarrhea, the presence of G. duodenalis can result in malabsorption; some studies have implicated this organism in decreased growth in some infected children and Last Updated: December 2012 possibly decreased productivity in young livestock. Outbreaks are occasionally reported in people, as the result of mass exposure to contaminated water or food, or direct contact with infected individuals (e.g., in child care centers). People are considered to be the most important reservoir hosts for human giardiasis. The predominant genetic types of G. duodenalis usually differ in humans and domesticated animals (livestock and pets), and zoonotic transmission is currently thought to be of minor significance in causing human illness. Nevertheless, there is evidence that certain isolates may sometimes be shared, and some genetic types of G. duodenalis (assemblages A and B) should be considered potentially zoonotic. Etiology The protozoan genus Giardia (Family Giardiidae, order Giardiida) contains at least six species that infect animals and/or humans. In most mammals, giardiasis is caused by Giardia duodenalis, which is also called G. intestinalis. Both names are in current use, although the validity of the name G. intestinalis depends on the interpretation of the International Code of Zoological Nomenclature.
    [Show full text]
  • The Cytoskeleton of Giardia Lamblia
    International Journal for Parasitology 33 (2003) 3–28 www.parasitology-online.com Invited review The cytoskeleton of Giardia lamblia Heidi G. Elmendorfa,*, Scott C. Dawsonb, J. Michael McCafferyc aDepartment of Biology, Georgetown University, 348 Reiss Building 37th and O Sts. NW, Washington, DC 20057, USA bDepartment of Molecular and Cell Biology, University of California Berkeley, 345 LSA Building, Berkeley, CA 94720, USA cDepartment of Biology, Johns Hopkins University, Integrated Imaging Center, Baltimore, MD 21218, USA Received 18 July 2002; received in revised form 18 September 2002; accepted 19 September 2002 Abstract Giardia lamblia is a ubiquitous intestinal pathogen of mammals. Evolutionary studies have also defined it as a member of one of the earliest diverging eukaryotic lineages that we are able to cultivate and study in the laboratory. Despite early recognition of its striking structure resembling a half pear endowed with eight flagella and a unique ventral disk, a molecular understanding of the cytoskeleton of Giardia has been slow to emerge. Perhaps most importantly, although the association of Giardia with diarrhoeal disease has been known for several hundred years, little is known of the mechanism by which Giardia exacts such a toll on its host. What is clear, however, is that the flagella and disk are essential for parasite motility and attachment to host intestinal epithelial cells. Because peristaltic flow expels intestinal contents, attachment is necessary for parasites to remain in the small intestine and cause diarrhoea, underscoring the essential role of the cytoskeleton in virulence. This review presents current day knowledge of the cytoskeleton, focusing on its role in motility and attachment.
    [Show full text]
  • Original Article Dientamoeba Fragilis Diagnosis by Fecal Screening
    Original Article Dientamoeba fragilis diagnosis by fecal screening: relative effectiveness of traditional techniques and molecular methods Negin Hamidi1, Ahmad Reza Meamar1, Lameh Akhlaghi1, Zahra Rampisheh2,3, Elham Razmjou1 1 Department of Medical Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran 2 Preventive Medicine and Public Health Research Center, Iran University of Medical Sciences, Tehran, Iran 3 Department of Community Medicine, School of Medicine, Iran University of Medical Sciences, Tehran, Iran Abstract Introduction: Dientamoeba fragilis, an intestinal trichomonad, occurs in humans with and without gastrointestinal symptoms. Its presence was investigated in individuals referred to Milad Hospital, Tehran. Methodology: In a cross-sectional study, three time-separated fecal samples were collected from 200 participants from March through June 2011. Specimens were examined using traditional techniques for detecting D. fragilis and other gastrointestinal parasites: direct smear, culture, formalin-ether concentration, and iron-hematoxylin staining. The presence of D. fragilis was determined using PCR assays targeting 5.8S rRNA or small subunit ribosomal RNA. Results: Dientamoeba fragilis, Blastocystis sp., Giardia lamblia, Entamoeba coli, and Iodamoeba butschlii were detected by one or more traditional and molecular methods, with an overall prevalence of 56.5%. Dientamoeba was not detected by direct smear or formalin-ether concentration but was identified in 1% and 5% of cases by culture and iron-hematoxylin staining, respectively. PCR amplification of SSU rRNA and 5.8S rRNA genes diagnosed D. fragilis in 6% and 13.5%, respectively. Prevalence of D. fragilis was unrelated to participant gender, age, or gastrointestinal symptoms. Conclusions: This is the first report of molecular assays to screen for D.
    [Show full text]
  • Catalogue of Protozoan Parasites Recorded in Australia Peter J. O
    1 CATALOGUE OF PROTOZOAN PARASITES RECORDED IN AUSTRALIA PETER J. O’DONOGHUE & ROBERT D. ADLARD O’Donoghue, P.J. & Adlard, R.D. 2000 02 29: Catalogue of protozoan parasites recorded in Australia. Memoirs of the Queensland Museum 45(1):1-164. Brisbane. ISSN 0079-8835. Published reports of protozoan species from Australian animals have been compiled into a host- parasite checklist, a parasite-host checklist and a cross-referenced bibliography. Protozoa listed include parasites, commensals and symbionts but free-living species have been excluded. Over 590 protozoan species are listed including amoebae, flagellates, ciliates and ‘sporozoa’ (the latter comprising apicomplexans, microsporans, myxozoans, haplosporidians and paramyxeans). Organisms are recorded in association with some 520 hosts including mammals, marsupials, birds, reptiles, amphibians, fish and invertebrates. Information has been abstracted from over 1,270 scientific publications predating 1999 and all records include taxonomic authorities, synonyms, common names, sites of infection within hosts and geographic locations. Protozoa, parasite checklist, host checklist, bibliography, Australia. Peter J. O’Donoghue, Department of Microbiology and Parasitology, The University of Queensland, St Lucia 4072, Australia; Robert D. Adlard, Protozoa Section, Queensland Museum, PO Box 3300, South Brisbane 4101, Australia; 31 January 2000. CONTENTS the literature for reports relevant to contemporary studies. Such problems could be avoided if all previous HOST-PARASITE CHECKLIST 5 records were consolidated into a single database. Most Mammals 5 researchers currently avail themselves of various Reptiles 21 electronic database and abstracting services but none Amphibians 26 include literature published earlier than 1985 and not all Birds 34 journal titles are covered in their databases. Fish 44 Invertebrates 54 Several catalogues of parasites in Australian PARASITE-HOST CHECKLIST 63 hosts have previously been published.
    [Show full text]
  • A New Species of Giardia Künstler
    Lyu et al. Parasites & Vectors (2018) 11:202 https://doi.org/10.1186/s13071-018-2786-8 RESEARCH Open Access A new species of Giardia Künstler, 1882 (Sarcomastigophora: Hexamitidae) in hamsters Zhangxia Lyu1,2†, Jingru Shao1†, Min Xue1, Qingqing Ye1, Bing Chen1, Yan Qin1,3 and Jianfan Wen1* Abstract Background: Giardia spp. are flagellated protozoan parasites that infect humans and many other vertebrates worldwide. Currently seven species of Giardia are considered valid. Results: Here, we report a new species, Giardia cricetidarum n. sp. in hamsters. Trophozoites of G. cricetidarum n. sp. are pear-shaped with four pairs of flagella and measure on average 14 μm (range 12–18 μm) in length and 10 μm (range 8–12 μm) in width. The trophozoites of the new species are generally larger and stouter than those of most of the other Giardia spp. and exhibit the lowest length/width ratio (c.1.40) of all recognized Giardia species. Cysts of G. cricetidarum n. sp. are ovoid and measure on average 11 μm (range 9–12 μm) in length and 10 μm (range 8–10 μm) in width and are indistinguishable from the cysts of other Giardia species. Molecular phylogenetic analyses based on beta-giardin, small subunit rRNA, and elongation factor-1 alpha loci all demonstrated that G. cricetidarum n. sp. is genetically distinct from all currently accepted Giardia spp. Investigation of the host range indicated that the new species was only found in hamsters (including Phodopus sungorus, P. campbelli and Mesocricetus auratus), while all the other described mammal-parasitizing species (G. muris, G.
    [Show full text]
  • Giardia Duodenalis in Children and Adults Attending a Day Care Centre in Central Italy Crotti D.*, D’Annibale M.L.*, Fonzo G.*, Lalle M.**, Cacciò S.M.** & Pozio E.**
    Article available at http://www.parasite-journal.org or http://dx.doi.org/10.1051/parasite/2005122165 DIENTAMOEBA FRAGILIS IS MORE PREVALENT THAN GIARDIA DUODENALIS IN CHILDREN AND ADULTS ATTENDING A DAY CARE CENTRE IN CENTRAL ITALY CROTTI D.*, D’ANNIBALE M.L.*, FONZO G.*, LALLE M.**, CACCIÒ S.M.** & POZIO E.** Summary: Résumé : LA PRÉVALENCE DE DIENTAMOEBA FRAGILIS EST PLUS ÉLEVÉE QUE CELLE DE GIARDIA DUODENALIS CHEZ LES ENFANTS ET LES ADULTES Giardia duodenalis is a well recognised enteropathogen, while EN TRAITEMENT AMBULATOIRE DANS L’ITALIE DU CENTRE Dientamoeba fragilis is rarely detected and consequently it is not recognised as an important human pathogen. In 2002-2003, a Giardia duodenalis est un parasite entérique bien connu, tandis survey has been carried out on enteroparasites in faecal samples que Dientamoeba fragilis, rarement détecté, n’est pas considéré of outpatients attending a day care centre in the town of Perugia comme un agent pathogène important chez l’homme. En 2002- (Central Italy). To improve the detection level, at least three 2003, une investigation sur les entéroparasites a été conduite sur samples from each patient were collected at different days and des échantillons de selles de patients en traitement ambulatoire within two hours from defecation. The coproparasitological dans la ville de Perugia (Italie du Centre). Afin d’augmenter le examination has been carried out by direct microscopic niveau de détection, un minimum de trois échantillons pour chaque examination, faecal concentration, and Giemsa and modified patient a été collecté aussitôt après la défécation et pendant Ziehl-Nielsen stainings of faecal smears. The genotypes of Giardia plusieurs jours.
    [Show full text]
  • Serosurvey and Molecular Detection of the Main Zoonotic Parasites Carried by Commensal Rattus Norvegicus Populations in Tehran, Iran
    Serosurvey and molecular detection of the main zoonotic parasites carried by commensal Rattus norvegicus populations in Tehran, Iran Taher Azimi ( [email protected] ) Tehran University of Medical Sciences https://orcid.org/0000-0003-0213-5227 Mohammad Reza Pourmand Tehran University of Medical Sciences Fatemeh Fallah Shaheed Beheshti University of Medical Sciences Abdollah Karimi Shaheed Beheshti University of Medical Sciences Roxana Mansour-Ghanaie Shaheed Beheshti University of Medical Sciences Seyedeh Mahsan Hoseini‐Alfatemi Shaheed Beheshti University of Medical Sciences Mehdi Shirdoust Shaheed Beheshti University of Medical Sciences Leila Azimi ( [email protected] ) Shaheed Beheshti University of Medical Sciences Research Keywords: Zoonotic parasites, Rattus norvegicus, Leishmania spp, Toxoplasma gondii, Giardia spp, Tehran Posted Date: March 28th, 2020 DOI: https://doi.org/10.21203/rs.3.rs-19380/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published at Tropical Medicine and Health on July 22nd, 2020. See the published version at https://doi.org/10.1186/s41182-020-00246-3. Page 1/10 Abstract Background: Rattus norvegicus are reservoirs of various zoonotic parasites that have become a global public health concern. Considering the distribution of Rattus norvegicus throughout Tehran, this study aims to assess the frequency of zoonotic parasites carried by commensal rodents in Tehran, Iran. Methods: The study considered ve regions (North, South, West, East, and center) of Tehran as case studies. The serological method was used for detecting antibodies against Trichomonas vaginalis , Babesia spp, and Cryptosporidium spp using a commercial qualitative rat ELISA kit.
    [Show full text]
  • Genomic and Functional Analysis of Viruses from Giardia Duodenalis Isolates
    biomedicines Article Re-Discovery of Giardiavirus: Genomic and Functional Analysis of Viruses from Giardia duodenalis Isolates Gianluca Marucci 1, Ilaria Zullino 1, Lucia Bertuccini 2 , Serena Camerini 2, Serena Cecchetti 2 , Agostina Pietrantoni 2, Marialuisa Casella 2, Paolo Vatta 1, Alex D. Greenwood 3,4 , Annarita Fiorillo 5 and Marco Lalle 1,* 1 Unit of Foodborne and Neglected Parasitic Disease, Department of Infectious Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy; [email protected] (G.M.); [email protected] (I.Z.); [email protected] (P.V.) 2 Core Facilities, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy; [email protected] (L.B.); [email protected] (S.C.); [email protected] (S.C.); [email protected] (A.P.); [email protected] (M.C.) 3 Leibniz Institute for Zoo and Wildlife Research, 10315 Berlin, Germany; [email protected] 4 Department of Veterinary Medicine, Freie Universität Berlin, 14195 Berlin, Germany 5 Department of Biochemical Science “A. Rossi-Fanelli”, Sapienza University, 00185 Rome, Italy; annarita.fi[email protected] * Correspondence: [email protected]; Tel.: +39-06-4990-2670 Abstract: Giardiasis, caused by the protozoan parasite Giardia duodenalis, is an intestinal diarrheal disease affecting almost one billion people worldwide. A small endosymbiotic dsRNA viruses, G. Citation: Marucci, G.; Zullino, I.; lamblia virus (GLV), genus Giardiavirus, family Totiviridae, might inhabit human and animal isolates Bertuccini, L.; Camerini, S.; Cecchetti, S.; Pietrantoni, A.; Casella, M.; Vatta, of G. duodenalis. Three GLV genomes have been sequenced so far, and only one was intensively P.; Greenwood, A.D.; Fiorillo, A.; et al.
    [Show full text]
  • Giardia Duodenalis in Environmental and Animal Samples in Scotland with Development of Novel
    Identification of Giardia duodenalis in Environmental and Animal Samples in Scotland with Development of Novel Approaches of Filtration Elution Ben Horton MSc by Research Institute of Biological Chemistry, Biophysics and Bio-engineering. Heriot-Watt University September 2016 “The copyright in this thesis is owned by the author. Any quotation from the thesis or use of any of the information contained in it must acknowledge this thesis as the source of the quotation or information." Abstract Giardia duodenalis is a waterborne flagellated protozoan parasite known to cause substantial cases of disease throughout the world. The parasite is argued to be zoonotic, and as such consumption of water contaminated by animal faeces containing parasite cysts is thought to lead to human infection. Infection is skewed towards the developing world, but outbreaks do occur within the developed world. This project had two aims: to identify the prevalence of G. duodenalis within a range of Scottish samples, both faecal and water, and to develop a novel method for the elution of G. duodenalis cysts from filter matrixes by incorporating megasonic sonication into a pre-existing method – the FiltaMax system. Molecular techniques found that water samples within this study were mostly negative for parasite DNA, however faecal samples were often positive, with animal samples testing sporadically positive throughout the study. A novel methodology for filter matrix elution of G. duodenalis cysts was developed and proven to be comparable to current leading filtration methods. This megasonic method also boasted significant advantages over the FiltaMax system: such as reduction in labour involved, substantially reduced damage of the parasite during elution and future automation is a possibility.
    [Show full text]
  • INFECTIOUS DISEASES of HAITI Free
    INFECTIOUS DISEASES OF HAITI Free. Promotional use only - not for resale. Infectious Diseases of Haiti - 2010 edition Infectious Diseases of Haiti - 2010 edition Copyright © 2010 by GIDEON Informatics, Inc. All rights reserved. Published by GIDEON Informatics, Inc, Los Angeles, California, USA. www.gideononline.com Cover design by GIDEON Informatics, Inc No part of this book may be reproduced or transmitted in any form or by any means without written permission from the publisher. Contact GIDEON Informatics at [email protected]. ISBN-13: 978-1-61755-090-4 ISBN-10: 1-61755-090-6 Visit http://www.gideononline.com/ebooks/ for the up to date list of GIDEON ebooks. DISCLAIMER: Publisher assumes no liability to patients with respect to the actions of physicians, health care facilities and other users, and is not responsible for any injury, death or damage resulting from the use, misuse or interpretation of information obtained through this book. Therapeutic options listed are limited to published studies and reviews. Therapy should not be undertaken without a thorough assessment of the indications, contraindications and side effects of any prospective drug or intervention. Furthermore, the data for the book are largely derived from incidence and prevalence statistics whose accuracy will vary widely for individual diseases and countries. Changes in endemicity, incidence, and drugs of choice may occur. The list of drugs, infectious diseases and even country names will vary with time. © 2010 GIDEON Informatics, Inc. www.gideononline.com All Rights Reserved. Page 2 of 314 Free. Promotional use only - not for resale. Infectious Diseases of Haiti - 2010 edition Introduction: The GIDEON e-book series Infectious Diseases of Haiti is one in a series of GIDEON ebooks which summarize the status of individual infectious diseases, in every country of the world.
    [Show full text]