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GIARDIA and CRYPTOSPORIDIUM Im ECJ Lo M in DOMESTIC GIARDIA AND CRYPTOSPORIDIUM im E C JlO m IN DOMESTIC LIVESTOCK: ZOONOTIC POTENTIAL, TRANSMISSION DYNAMICS, AND THREAT TO DRINKING WATER A Thesis Submitted to the Graduate Faculty In Partial Fulfilment of the Equirements For the Degree of Doctor of Philosophy In the Department of Health Management Faculty of Veterinary Medicine University of Prince Edward Island Ebo Budu-Amoako Charlottetown, P. E. 1. April, 2011 2011. E. Budu-Amoako Library and Archives Bibliothèque et Canada Archives Canada Published Heritage Direction du Branch Patrimoine de l’édition 395 Wellington Street 395, rue Wellington Ottawa ON K1A 0N4 Ottawa ON K1A 0N4 Canada Canada Your file Votre référence ISBN: 978- 0 - 494- 82269-2 Our file Notre référence ISBN: 978- 0- 494- 82269-2 NOTICE: AVIS: The author has granted a non­ L’auteur a accordé une licence non exclusive exclusive license allowing Library and permettant à la Bibliothèque et Archives Archives Canada to reproduce, Canada de reproduire, publier, archiver, publish, archive, preserve, conserve, sauvegarder, conserver, transmettre au public communicate to the public by par télécommunication ou par l'Internet, prêter, telecommunication or on the Internet, distribuer et vendre des thèses partout dans le loan, distribute and sell theses monde, à des fins commerciales ou autres, sur worldwide, for commercial or non­ support microforme, papier, électronique et/ou commercial purposes, in microform, autres formats. paper, electronic and/or any other formats. The author retains copyright L’auteur conserve la propriété du droit d’auteur ownership and moral rights in this et des droits moraux qui protège cette thèse. Ni thesis. Neither the thesis nor la thèse ni des extraits substantiels de celle-ci substantial extracts from it may be ne doivent être imprimés ou autrement printed or otherwise reproduced reproduits sans son autorisation. without the author’s permission. In compliance with the Canadian Conformément à la loi canadienne sur la Privacy Act some supporting forms protection de la vie privée, quelques may have been removed from this formulaires secondaires ont été enlevés de thesis. cette thèse. While these forms may be included Bien que ces formulaires aient inclus dans in the document page count, their la pagination, il n’y aura aucun contenu removal does not represent any loss manquant. of content from the thesis. I 4 'l Canada CONDITION OF USE The author has agreed that the Library, University of Prince Edward Island, may make this thesis freely available for inspection. Moreover, the author has agreed that permission for extensive copying of this thesis for scholarly purposes may be granted by the professor or professors who supervised the thesis work rcorded herein or, in their absence, by the Chairman of the Department or the Dean of the Faculty in which the thesis work was done. It is understood that due recognition will be given to the author of this thesis and to the University of Prince Edward Island in any use of the material in this thesis. Copying or publication or any other use of the thesis for financial gain without approval by the University of Prince Edward Island and the author’s written permission is prohibited. Request for permission to copy to make any other use of material in this thesis in whole or in part should be addressed to: Chair of the Department of Elealth Management Faculty of Veterinary Medicine University of Prince Edward Island Charlottetown, P. E. I. Canada, CIA 4P3 11 SIGNATURE PAGE ( fii) ^ a y) REMOVED Abstract The main aims of this thesis were to obtain information on the prevalence of zoonotic and host-specific Cryptosporidium and Giardia in livestock on Prince Edward Island, Canada, to determine if the presence of cattle and pigs increase the levels of Giardia and Cryptosporidium in source water. Giardia genotypes and Cryptosporidium species in humans on PEI were also analysed to determine if they were the same as those found in cattle and pigs. The role that dairy farms may play in contaminating source water on Prince Edward Island, Canada, with Giardia and Cryptosporidium is described in Chapter 2. Thirty-two percent and 14% of 752 fecal samples, and 100 and 55% of 20 herds, were positive for Giardia and Cryptosporidium, respectively. Most of theCryptosporidium isolates belonged to C. andersoni and C. bovis. Giardia duodenalis assemblage E was predominant among all cattle age groups, while only 6 and 4% of isolates belonged to assemblages A and B, respectively. Giardia and Cryptosporidium were detected in 46 and 85% of surface water samples, respectively, while one groundwater sample was positive for Cryptosporidium. All four genotyped Cryptosporidium isolates from water were C.parvum. This study indicates that dairy cattle were predominantly infected with non-zoonotic species/genotypes ofCryptosporidium and Giardia. However, some surface water sources were contaminated with the zoonotic C.parvum and may represent a threat to the watershed and subsequent disease transmission to humans. Cattle >6 months of age were less likely to be infected with Giardia while there was no significant association between either the location of the farm or the housing type and infection withGiardia. Cattle >6 months of age in free-stall housing were more likely to be infected with Cryptosporidium species. While there was no impact of age, infection withCryptosporidium tended to be higher in Queens County. Chapter 3 describes the prevalence and genotypes of Giardia and Cryptosporidium on beef farms, including the water sources associated with the farms, and the risk factors for infection of cattle with these parasites. Giardia and Cryptosporidium were detected in 42% and 17% of 739 fecal samples, and 100 and 80% of 20 farms, respectively. G.duodenalis livestock genotype assemblage E predominated (89%). The zoonotic assemblages A and B were found in 4 and 7% of the fecal samples that were genotyped, respectively. Overall, the most common Cryptosporidium species detected in this study was C. andersoni. Giardia cysts and Cryptosporidium oocysts were detected in 14 and 93% of surface water samples of 14 farms, respectively. Cryptosporidium oocysts were detected in three (15%) ground water samples of 20 farms. Age of animals and location of the farm, were not associated with the risk of infection in cattle with either Cryptosporidium spp. or G. duodenalis. The presence of zoonotic species of Cryptosporidium and Giardia in beef eattle represents a potential threat to source water and subsequent disease transmission to humans from these parasites In Chapter 4, the prevalence of infection with Cryptosporidium and Giardia, and the genotypes and species of isolates, were determined in order to establish the zoonotic potential of pigs. Eighteen herds (86%) and 163 animals (26%; 95% Cl: 22-29%) tested positive for Cryptosporidium, while just 3 herds (14%) and 6 animals (1%; 95% Cl: 0.4-2%) tested positive for Giardia. The Cryptosporidium spp. most isolated among those successfully genotyped, belonged to pig genotype 11 (61%) and C. suis (36%). Pigs are not a major source ofGiardia and the common occurrence of the host-specific genotypes and species Cryptosporidium,of indicate that domestic pigs, likely do not pose a significant health risk to humans from these parasites. The species and genotypes ofCryptosporidium and Giardia in livestock were compared with those from humans to determine if zoonotic transmission of these species occurred, in VI Chapter 5. Overall 22% and 0.5% of human fecal samples were positive for Cryptosporidium and Giardia respectively. The species ofCryptosporidium found in humans were C.parvum and C hominis. C.parvum shedding in humans was more frequent than C. hominis. Sub-genotyping of C. parvum isolates from beef cattle revealed three sub-genotypes: IIaA16G2Rl, IIaA16G3Rl and IIaA15G2Rl. Two of these sub-genotypes: IIaA16G2Rl and IIaA15G2Rl were also found in humans. The results indicate that prevalences of the zoonotic species and genotypes of Cryptosporidium and Giradia in the herds sampled in this study were low. However, the data suggests zoonotic transmission ofCryptosporidium to humans on PEI and that cattle are the possible source of the parasite. V ll Acknowledgements I would like to thank my supervisors, Drs. J McClure and Spencer Greenwood for their invaluable support and kindness in guiding me through this whole journey. I am particularly indebted to Herman Barkema, my former supervisor for having the confidence in me to accept me as a graduate student to work on this project. My supervisory committee members Drs. Spencer Greenwood, J McClure, Dan Humik, Brent Dixon, Herman Barkema, Lamont Sweet and Lei Ang/Lewis Abbot deserve my utmost thanks for their guidance throughout my thesis. I would particularly like to express my appreciation to Spencer for his invaluable assistance and encouragement when things seemed hopeless. I am grateful to Drs. Henrik Stryhn, Alejandro Ceballos, Elmabrok Masaoud and Javier Sanchez for their invaluable assistance with the data analysis. I would like to express my appreciation to Cynthia Mitchell, Mathew Saab, Loma Farrington and various summer veterinary students for technical support. This project would not have been possible without the financial support from Health Canada. I am also grateful to the Prince Edward Island Food Technology Centre for the financial support and time provided to me. Finally, I would like to express my utmost thanks and appreciation to my family, to whom the thesis is dedicated, for their
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