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Comparison of Naturally-Acquired and Vaccine-Acquired Immunity to Loma Salmonae in Rainbow Trout (Oncorhynchus Mykiss)

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Comparison of Naturally-Acquired and Vaccine-Acquired Immunity to Loma Salmonae in Rainbow Trout (Oncorhynchus Mykiss) COMPARISON OF NATURALLY-ACQUIRED AND VACCINE-ACQUIRED IMMUNITY TO LOMA SALMONAE IN RAINBOW TROUT (ONCORHYNCHUS MYKISS) BY JENNIFER E. HARKNESS A Thesis Submitted to the Graduate Faculty in Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE Department of Pathology and Microbiology Faculty of Veterinary Medicine University of Prince Edward Island ©APRIL 2012. J.E. HARKNESS Library and Archives Bibliotheque et Canada Archives Canada Published Heritage Direction du 1+1Branch Patrimoine de I'edition 395 Wellington Street 395, rue Wellington Ottawa ON K1A0N4 Ottawa ON K1A 0N4 Canada Canada Your file Votre reference ISBN: 978-0-494-94063-1 Our file Notre reference ISBN: 978-0-494-94063-1 NOTICE: AVIS: The author has granted a non­ L'auteur a accorde une licence non exclusive exclusive license allowing Library and permettant a la Bibliotheque 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 telecommunication ou par I'lnternet, preter, telecommunication or on the Internet, distribuer et vendre des theses partout dans le loan, distrbute and sell theses monde, a des fins commerciales ou autres, sur worldwide, for commercial or non­ support microforme, papier, electronique et/ou commercial purposes, in microform, autres formats. paper, electronic and/or any other formats. The author retains copyright L'auteur conserve la propriete du droit d'auteur ownership and moral rights in this et des droits moraux qui protege cette these. Ni thesis. Neither the thesis nor la these ni des extraits substantiels de celle-ci substantial extracts from it may be ne doivent etre imprimes ou autrement printed or otherwise reproduced reproduits sans son autorisation. without the author's permission. In compliance with the Canadian Conformement a la loi canadienne sur la Privacy Act some supporting forms protection de la vie privee, quelques may have been removed from this formulaires secondaires ont ete enleves de thesis. cette these. 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. Canada CONDITIONS FOR THE USE OF THE THESIS 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 recorded herein or, in their absence, by the Chair 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. Requests for permission to copy or to make any other use of material in this thesis in whole or in part should be addressed to: Chair of the Department of Pathology and Microbiology Faculty of Veterinary Medicine University of Prince Edward Island Charlottetown, P. E. I. Canada CIA 4P3 SIGNATURE PAGE AV/J* (w) REMOVED ABSTRACT Microsporidial Gill Disease of Salmon (MGDS) is a result of infection withLoma salmonae and has been described in fanned and wild Pacific salmon as causing severe branchitis, resulting in significant economic losses in aquaculture, as fish succumbing to the disease are those nearing market size. No treatment currently exists to controlL. salmonae, although previous studies have documented disease resistance upon initial recovery and secondary exposure to the parasite. The objectives of this study were to further examine naturally acquired as well as vaccine-acquired immunity toL. salmonae in rainbow trout (RBT). It is believed that farm reared salmon are exposed to continual low doses of the parasite over time, and the naturally acquired immunity studies focused on the protective response generated by recovery of a low dose cohabitation exposure and subsequent high dose oral re-challenge. Having the ability to partially protect RBT upon re-exposure toL. salmonae contradicted what is being seen in the netpen scenario, as Chinook and Coho salmon are assumed to have no immunity to re-infection. The investigation into vaccine-acquired immunity focused on the use of heterologous live vaccines, containing Glugea anomala and Glugea hertwigi, respectively. Having common characteristics and being phylogenetically related L.to salmonae, the intraperitoneal (IP) injection into RBT 6 weeks pre­ exposure toL. salmonae was able to reduce xenoma burden significantly as compared to naive controls. Since live vaccines are not marketable, further studies investigated the use of killed heterologous (G. anomala and G. hertwigi) vaccines in comparison with the widely published killed homologous vaccine containing Loma sp. The superiority of both the killed heterologous vaccines were noted, as they were able to reduce xenoma burden upon exposure to the parasite by upwards of 15% compared to the killed homologous vaccine. As a final approach, naturally- acquired immunity was revisited to determine if prophylactic oral treatment of immunostimulant, Provale™, a 0-1,3/1,6 glucan in synergy with low dose recovery could significantly boost the immune response of RBT toL. salmonae. Little evidence was found to support this hypothesis. The immune responses and consequently protection to MGDS in RBT depends greatly upon the intensity of the primary infection and subsequent recovery. Timing of vaccination is essential to the protection of RBT to L. salmonae. Although RBT is a useful surrogate model for infection with MGDS, it does not mimic the exact severity of infection seen in Pacific salmon, particularly Chinook and Coho salmon. With this in mind, experimental studies mimicking those carried out in the thesis in Chinook salmon would be beneficial to the control and treatment of L. salmonae. v ACKNOWLEDGEMENTS I wish to thank my supervisor Dr. David Speare for his constant support, creativity, sincerity and encouragement throughout my program. I greatly respect your enthusiasm as a research scientist and supervisor and appreciate your honesty and personal integrity. I would also like to thank the other members of my Supervisory Committee, Drs. Fred Markham, Spencer Greenwood, Collins Kamunde, Fred Kibenge and the members of my Examination Committee, Drs. David Speare, Gary Conboy, Collins Kamunde, Mark Fast and Spencer Greenwood for reviewing and correcting my thesis. I would still be wandering around the Aquatic Facility staring at empty tanks without the support and encouragement of Dr. Nicole Guselle. As you can imagine, she has had a significant hand in the initialization and continuation of all the experimental aspects of the thesis. In addition, many thanks and gratitude to the AVC Aquatic Facility staff including Wayne Petley, Angela Hamish- Driscoll and Lee Dawson, I couldn’t have achieved this without your tireless work ethics to maintain an amazing facility. Thanks to Diane MacLean and Rita Saunders for their administrative assistance and Kathy Jones for her photography assistance. I am extremely grateful to have met so many interesting people over the past few years and wish to thank everyone who has had a hand in the completion of this thesis, whether it was through guidance, having a shoulder to cry on, listening to weekly, if not daily rants of a graduate student, or simply being there in a time of need. To each and every one of you, I thank you for your kindness, support, friendship and love. Thanks to my parents Donna McKinnon and Michael Poczynek, to my sister Sarah and to the rest of my family for their continued support, encouragement, constructive criticism, and ultimately love. You all hold a special place in my heart. Special thanks goes to my dearest of friends, Jessica Fry and Angela Hamish-Driscoll. You have both made me a better person and I love you both dearly. A very large, and very deserving thanks goes to the hundreds of fish that were used in my research, because without them, this thesis would have never been possible. Their sacrifice is truly appreciated and will serve the greater good of the future of aquaculture. I acknowledge the financial support I received through a Natural Sciences and Engineering Research Council Strategic Grant, the Springboard Atlantic Fund and the Department of Pathology and Microbiology at the Atlantic Veterinary College (UPEI). DEDICATION To my grandparents, Della and Burleigh MacKinnon. Grammy, I hope I’ve made you proud! Grampy, your love of fish and wildlife resonates in me! Thank you. Love Always, Jenny Penny vii TABLE OF CONTENTS Title of Thesis i Conditions of Use ii Permission to Use Postgraduate Thesis iii Certification of Thesis Work iv Abstract v Acknowledgements vi Dedication vii Table of Contents viii-x List of Figures xi List of Tables xii List of Abbreviations xiii 1. GENERAL INTRODUCTION 1.1 Aquaculture 1 1.1.1 Global Production 1 1.1.2 Canadian Production 2 1.1.3 Salmonid Production 4 1.1.4 Chinook Salmon 5 1.1.5 Netpens 7 1.2 Disease in Netpens 9 1.2.1 Diseases of Chinook Salmon 10 1.2.2 Protistan Diseases of Salmon in Seawater 11 1.3 Microsporidian Parasites 13 1.3.1 Microsporidian Life Cycle 15 1.3.2 Microsporidians in Human and Animal Health 18 1.4 Loma salmonae 20 1.4.1 Transmission 21 1.4.2 Diagnosis and Clinical Signs 22 1.4.3 Immunological Resistance 26 1.4.4 Drug Treatment 29 1.4.5 Vaccines Against Loma salmonae 32 1.5 Research Objectives 33 1.6 REFERENCES 35 2.
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