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Development of the Marine Myxozoan, Kudoa Thyrsites

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Development of the Marine Myxozoan, Kudoa Thyrsites DEVELOPMENT OF THE MARINE MYXOZOAN, KUDOA THYRSITES (GILCHRIST, 1924), IN NETPEN-REARED ATLANTIC SALMON (SU0 SAUR L.) IN BRlTISH COLUMBIA Jonathan David William Moran B-Sc., University of New Brunswick (Fredericton), 1992 M-Sc., University of New Brunswick (Fredericton), 1994 A THESIS SUBMIïTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in the Department of Biological Sciences O Jonathan D.W. Moran 1999 SIMON FRASER UNIVERSITY May 1999 Al1 rights resewed. This work rnay not be reproduced in whole or in part, by photocopy or other means, without permission of the author. National Library Bibliothèque nationale 1*1 of Canada du Canada Acquisitions and Acquisitions et Bibliographie Services services bibliographiques 395 Wellington Street 395. me WeHingtm OttawaON K1A ON4 OcrawaON KiA OF)4 Canada Canada The author has granted a non- L'auteur a accordé une licence non exclusive Licence aliowing the exclusive permettant à la National Libras, of Canada to Bibliothèque nationale du Canada de reproduce, loan, distribute or seii reproduire, prêter, distribuer ou copies of this thesis in microfom, vendre des copies de cette thèse sous paper or electronic formats. la forme de microfiche/film, de reproduction sur papier ou sur format électronique. The author retains ownership of the L'auteur conserve la propriété du copyright in this thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantial extracts fiom it Ni Is thkse ni des extraits substantiels may be printed or othenvise de celle-ci ne doivent être imprimés reproduced without the author' s ou autrement reproduits sans son permission. autorisation. ABSTRACT firdoa rhyrsites (phylum Myxozoa) is of concern to the British Columbia (BC) aquaculture industry because of its association with pst-rnortem myoliquefaction in netpen-reared Atlantic salmon (Sdmo salar L.). This condition, commonly referred to as soft flesh syndrome. significantly decreases the market value of afTected fish products. The seasonality of K. thyrsites was investigated following natural exposure of Atlantic saImon in seawater netpens at the Pacific Biologicai Station, in Nanaimo. BC. Atlantic salmon were exposed for several 8-week penods throughout the year, and it was determined that the infections were contracted in only the summer and Ml. The progression of these infections in both netpen and tank popuiations was followed for up to 20 months. The majority of fish contracted K. rhyrsites infections. Sporulation typically occurred within 6 months (approximately 2000 degree-days) afier transfer to seawater, and the fish almost completely recovered fiom the original infections within a year. Of 3 1 nonsalmonid fish species collected fiom the vicinity of seawater netpens and fiom research cruises off Vancouver Island, five species were identified as potentiai reservoir hosts incl uding rock sole (Pleuronectes bilineatus), tube-snout (Aulorhynchusflavidus), arrowtooth flounder (Atheresthes sromias), and two new host records, lingcod (Ophiodon elongatus) and thread fin sculpin (Icelinusfilamentosus). Attempts to transmit the parasite directly fiom fish to fish by intubation of fiesh myxospores were unsuccessfûi. However, intraperitoneal injection of blood collected from a coho salmon (Oncorhynchus kisutch) infected with K. thyrsites successfully transmitted the infection. Using light rnicroscopy to investigate the sequential development, K. rhyrsites infections were detected in the somatic musculature at 13 weeks pst-exposure (pe.). In a subsequent experiment, infections were first detected at 6 weeks p-e- using a PCR test and afier 9 weeks p.e. using light microscopy. The earliest developmental stage detected by histology was a smail plasmodium containhg four nuclei at 9 weeks p.e. No host response was observed histologicaily that was directly related to muscle fibers which contained intact pseudocysts. However, the response associated with ruptured pseudocysts was characterized by chronic, multifocal inflammation between the muscle fibers. DEDICATION This dissertation is dedicated to my farnily, and especially my parents, who have provided me with unwavering support throughout my university career. This accomplishment would not have been possible without their guidance and moral support. 1 would also like to dedicate this, in part, to the residents of St. Martins, New Brunswick, many of whom have had a significant impact on my values and beliefs. Al1 men dream: but not equally. Those who drearn by night in the dusty recesses of their minds wake in the day to find that it wris vanity: but the drearners of the day are dangerous men. for they may act their dream with open eyes. to make it possible. T.E. Lawrence ACKNOWLEDGEMENTS 1 wouid like to express my sincere thanks to rny academic supervisor. J.M. Webster? for his encouragement throughout this study. His financial support for travel to national and international conferences is laudable. My complete gratitude goes to my research supervisor. M.L. Kent? without whose enthusiastic assistance and financial support. this project would never have been able to continue for the past five years. His unconditional moral support, in times when 1 doubted both myself and others, kept me headed in the right direction. 1 must also acknowledge my bnef, but priviledged, period with L. Margolis. His sudden passing was certainly a somber event during my tenure at the Pacific Biologicai Station (PBS). I wouId like to acknowledge the role of several employees of the PBS, without whom this research would not have been able to take place on such a scale. In particular, 1 must thank the director of the PBS, D. Noakes, for allowing me access to the PBS facilities for the past 4 years. Also, the assistance of the fish health and histology section employees is gratefully acknowledged, especially S. Dawe, D. Whitaker, T. McDonald, J. Richard. J. Bagshaw, and V. Rantis. Laboratory space and technicai assistance with the molecular screening were provided by R.H. Devlin and J. Khattra. Fish maintenance at the experimental farm was performed by R. Kennedy and L. Lewington. Of no iess importance was R. Kennedy's organization of the annual hockey pool. Scholarships and fellowships were provided by the Science Council of British Columbia and Simon Fraser University. Research funding was provided by Pacific Aqua Salmon Farming Partnerships (l3. Hicks), Fisheries and Oceans Canada, and the Natural vii Sciences and Engineering Research Council. 1 would also like to thank M. Bhakthan and S. Foran of Simon Fraser University for diligently forwarding mail to me in Nanaîmo, and keeping me informed of pertinent University news- Permission to reproduce photographs was provided by S. Hallett (University of Queensland), Y. Maeno (National Research Institute of Aquaculture Fisheries Agency), and H. Yokoyama (University of Tokyo). Kudoo rhyrsires fiom South Afiica was provided by M. Kerstan of the Sea Fisheries Research Institute. The English translation of Egusa (1 986) was provided by S. Feist (Centre for Environment, Fisheries & Aquaculture Science (CEFAS)). I would also like to acknowledge the hospitality of S. Feist and M. Longshaw during my visit to CEFAS. viii TABLE OF CONTENTS Page ... ABSTRACT ............................................................................................................. .IH DEDICATION .......................................................................................................... v QUOTATION ...................................................................................................... .vi .. ACKNO WLEDGEMENTS .......................................... vu LIST OF TABLES .................................................................................................... xiv LIST OF PLATES AND FIGWS ......................................................................... xv CHAPTER 1 - INTRODUCTION ........................................................................... 1 Salmonid aquaculture in British Columbia ............................................. 2 Effect of fish pathogens on the BC aquaculture ïndustry ...................... .4 Drug treatments ....................................................................................... 7 Vaccines ................................................................................................. .7 The phylum Myxozoa Grassé, 1970 ................... ..... ............................ 8 Life cycle and development of myxosporeans ........................................ 1 1 The genus Kudoa Meglitsch, 1947 ......................................................... 15 Kudoa thyrsires and the BC aquaculture industry ................................... 16 Objectives of this study ......................................................................... 16 CHAPTER 2 - GENERAL METHODS .................................................................-1 8 1. LocaIity ................................................................................................... 19 2. Holding facilities ..................................................................................... 19 Page 2.1. Seawater netpens ...................... .,., ................................................... 19 2.2. Freshwater tanks .............................................................................. 19 2.3. Seawater tanks ................................................................................. 19 3 . Hosts ......................................................................................................
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