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Factors Affecting the Resistance of Juvenile Rainbow Trout to Whirling

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Factors Affecting the Resistance of Juvenile Rainbow Trout to Whirling Factors affecting the resistance of juvenile rainbow trout to whirling disease by Eileen Karpoff Nicole Ryce A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Fish and Wildlife Biology Montana State University © Copyright by Eileen Karpoff Nicole Ryce (2003) Abstract: Whirling disease has been linked to significant losses of rainbow trout in wild trout populations. I examined the relationships among the whirling disease pathogen (Myxobolus cerebralis), its hosts, and the environment, with emphasis on factors affecting the resistance of rainbow trout to whirling disease. I first determined the effects of age on the development of whirling disease in rainbow trout. Whirling disease was substantially reduced when the fish were exposed to the parasite for the first time at 9 weeks posthatch (756 degree-days) or older. Second, I determined if the relationship of increasing age at exposure causing a reduction in disease severity was a factor of age or size of fish at exposure. I demonstrated that both the age of fish at first exposure and the size of fish at first exposure were important for the development of whirling disease. Third, I determined if rainbow trout can develop an acquired immune response to M. cerebralis and whether it provides the fish with protection against subsequent exposures. Rainbow trout did develop an acquired immune response to the parasite and an initial immunization exposure provided the fish with protection against subsequent exposures. However, the immunization, which provided the fish with protection, also induced disease in the fish, which outweighed the benefits of the protection. To reduce the effects of whirling disease on rainbow trout, they should be reared in M. cerebralis-free waters for 756 degree-days of development or until they are 40 mm in length, whether in the wild or in a hatchery situation. FACTORS AFFECTING THE RESISTANCE OF JUVENILE RAINBOW TROUT TO WHIRLING DISEASE by Eileen KarpoffNicol Ryce A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Fish and Wildlife Biology MONTANA STATE UNIVERSITY Bozeman, Montana January 2003 11 OlIS RAW APPROVAL of a dissertation submitted by Eileen KarpoffNicol Ryce This dissertation has been read by each member of the dissertation committee and has been found to be satisfactory regarding content, English usage, format, citations, bibliographic style, and consistency, and is ready for submission to the College of Graduate Studies. Alexander V. ZaIe "5X7 //L-- » 13 OS Cljaifperson,vGradu£(f^ Committee Date Approved for the Department of Ecology Jay J. Rotella g^paifment Head, Ecology Department Date Approval for the College of Graduate Studies Bruce R. McLeod Graduate Dean Date STATEMENT OF PERMISSION TO USE In presenting this dissertation in partial fulfillment of the requirements for a . doctoral degree at Montana State University, I agree that the Library shall make it available to borrowers under rules of the Library. I further agree that copying of this dissertation is allowable only for scholarly purposes, consistent with “fair use” as prescribed in the U.S. Copyright Law. Requests for extensive copying or reproduction of this dissertation should be referred to Bell and Howell Information and Learning, 300 North Zeeb Road, Ann Arbor, Michigan 48106, to whom I have granted “the exclusive right to reproduce and distribute my dissertation in and from microform along with the non-exclusive right to reproduce and distribute my abstract in any format in whole or in part.” ACKKNOWLEDGMENTS I thank the Whirling Disease Initiative of the National Partnership on Management of Wild and Native Coldwater Fisheries for providing funding to complete this work. I express my greatest appreciation to Cal Fraser, Laboratory Manager of the Wild Trout Research Laboratory, for his endless technical assistance. Sincerest thanks and appreciation go to the many technicians who provided invaluable help on this project over the years. Mark Nelson, Doug Rider, Terah Jentzsche, Andrea Gray, Chris Singer, Chris Kohler, Seth Judge, Amy Thompson, Ryen Anderson, Jeanne Knox, Molly Toner, Robin Stevenson, Amy Herrera, Andy Smith and Ty Wyatt all provided countless hours of assistance. This project would not have been possible without their hard work and dedication. The moral support and help provided to me by my fellow graduate students is also greatly appreciated, especially Matt Jaeger, Doug Rider, Andrew Munro and Lee Bergstedt for tolerating me as their office mate. I thank Ron Hedrick and Mark Adkison at the University of California, Davis, for allowing me to use their laboratory and expertise, which made the acquired immunity portion of this work possible. My co-advisors Alexander Zale and Elizabeth MacConnell, and my graduate committee, deserve special recognition for their endless guidance, support and patience throughout the years. Dee Topp is also acknowledged for her invaluable administrative support and friendship in the Montana Cooperative Fisheries Research Unit. Finally, I acknowledge my mum, dad, and husband, without their support, love and encouragement I would never have completed this work. V TABLE OF CONTENTS LIST OF TABLES....................................................................................................... vii LIST OF FIGURES..................................................................................................... viii ABSTRACT.................................................................................................................... xvi 1. INTRODUCTION................................................................................................. I 2. OBJECTIVE I : EFFECTS OF RAINBOW TROUT AGE AND PARASITE DOSE ON THE DEVELOPMENT OF WHIRLING DISEASE......................... 10 Introduction.................................................................................................... 10 Methods............................................................................................................... 13 Experimental Procedures............................................................................... 14 Statistical Analysis........................................................................................ 19 Results................................................................................................................. 22 Cumulative Mortality.................................................................................... 22 Swimming Performance............................................ 24 Microscopic Pathology.................................................................................. 24 Spore Counts................................................................................................ 29 Clinical Signs................................................................................................. 31 Discussion............................................................................................................ 39 3. OBJECTIVE 2: EFFECTS OF AGE VERSUS SIZE AT TIME OF EXPOSURE ON THE DEVELOPMENT OF WHIRLING DISEASE IN RAINBOW TROUT.............................................................................................. 56 Introduction....................................................................................................... 56 Methods............................................................................................................... 59 Experimental Procedures................................................................................ 59 Statistical Analysis......................................................................................... 65 Results................................................................................................................. 68 Cumulative Mortality.................................................................................... 68 Swimming Performance................................................................................ 68 Clinical Signs................................................................................................. 70 Microscopic Pathology.................................................................................. 75 Spore Counts.......................................................................................... 75 Cartilage......................................................................................................... 78 Discussion......................................................................................... 83 vi TABLE OF CONTENTS - CONTINUED 4. OBJECTIVE 3: EFFECTS OF AN INITIAL EXPOSURE TO MYXOBOL US CEREBRALIS, AND INDUCTION OF AN ACQUIRED IMMUNE RESPONSE, ON THE DEVELOPMENT OF RESISTANCE TO WHIRLING DISEASE IN RAINBOW TROUT.................................................................... 91 Introduction....................................................................................................... 91 Methods.................. 97 Experimental Procedures.......................................................................... 97 Statistical Analysis............ .................................................. 103 Results............................................................................. 107 Disease indicators.............................................................................................. 107 Serum antibody levels: 9-week posthatch exposure........................................
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