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Comparative Effects of the Toxic Dinoflagellate, Karenia Brevis, on Bivalve Molluscs from Florida James R

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Comparative Effects of the Toxic Dinoflagellate, Karenia Brevis, on Bivalve Molluscs from Florida James R University of South Florida Scholar Commons Graduate Theses and Dissertations Graduate School 2007 Comparative effects of the toxic dinoflagellate, Karenia brevis, on bivalve molluscs from Florida James R. Leverone University of South Florida Follow this and additional works at: http://scholarcommons.usf.edu/etd Part of the American Studies Commons Scholar Commons Citation Leverone, James R., "Comparative effects of the toxic dinoflagellate, Karenia brevis, on bivalve molluscs from Florida" (2007). Graduate Theses and Dissertations. http://scholarcommons.usf.edu/etd/2260 This Dissertation is brought to you for free and open access by the Graduate School at Scholar Commons. It has been accepted for inclusion in Graduate Theses and Dissertations by an authorized administrator of Scholar Commons. For more information, please contact [email protected]. Comparative Effects of the Toxic Dinoflagellate, Karenia brevis, on Bivalve Molluscs from Florida by James R. Leverone A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy Department of Biological Oceanography College of Marine Science University of South Florida Major Professor: Norman J. Blake, Ph.D Sandra E. Shumway, Ph.D, D.Sc. Gabriel A. Vargo, Ph.D. Joseph J. Torres, Ph.D. Richard H. Pierce, Ph.D. Date of Approval: April 2, 2007 Keywords: harmful algae, molluscs, larvae, juveniles, mortality, clearance rate, histopathology © Copyright 2007, James R. Leverone Dedication This dissertation is wholeheartedly dedicated to my gracious wife, Barbara. Throughout my graduate studies, she has steadfastly supported me through all my trials and tribulations. Whenever an experimental procedure went awry, especially in the beginning, she was always there to put things in perspective, reminding me that through apparent failures, progress and learning took root. And at times when I was not making even the slightest progress or was not paying proper attention to my work, she would gently push me to keep going. Even at times when I had doubts, she never wavered. She had complete trust and confidence in me at times when I questioned myself. Furthermore, she always maintained a convivial home and family life which provided a welcome respite from my studies. I will be forever grateful to her for her support and hope she may share in the joy of my accomplishment. May this work be an inspiration to any student contemplating a return to graduate school later in life. May you have a partner who will encourage and support you as my wife did me. Acknowledgments There are many people to whom I owe a debt of appreciation for their support during the pursuit of this dissertation. I wish to thank Dr. Norman J. Blake and Dr. Sandra E. Shumway for their guidance throughout this process, as well as my remaining committee members, Drs. Gabe Vargo, Jose Torres and Rich Pierce. Candice Way and Noland Elsaesser at USF provided cultured bay scallop larvae and juveniles. Curt Hemmel of Bay Shellfish Company supplied quahog larvae and juveniles, while John Suppan was very gracious in providing oyster larvae. Many personnel from Mote Marine Laboratory were truly remarkable in extending their support and encouragement. Dana Wetzel provided valuable climate-controlled laboratory space. Mike Henry and Trish Blum extracted toxins from numerous algal cultures and performed brevetoxin analyses; Chris Higham and Val Palubok maintained phytoplankton cultures. Jim Gelshleiter helped me acquire a modicum of skill in histological techniques and absorbed the cost laboratory supplies. I am very honored to have had the continual support of Dr. Ernest Estevez, Director of the Center for Coastal Ecology, as I attempted to juggle my dual role of staff biologist at Mote Marine Laboratory and student at the University of South Florida. Dr. Estevez maintained enthusiastic support and encouragement for my academic quest. Finally, this degree could not have been completed without the support and financial assistance of the president, Dr. Kumar Mahadevan, and board of directors of Mote Marine Laboratory. I am deeply appreciative and truly thankful to all of you. Table of Contents List of Tables .................................................................................................................... iii List of Figures.................................................................................................................... v Abstract........................................................................................................................... viii Chapter One: Introduction ................................................................................................. 1 Chapter Two: Literature Review ....................................................................................... 4 Bivalve Mortality Associated with Harmful Algal Species.................................... 5 Behavioral Responses of Bivalves to Harmful Algal Species.............................. 10 Feeding Responses of Bivalves Exposed to Harmful Algal Species.................... 12 Cytotoxic Effects of Harmful Algal Species to Bivalves ..................................... 20 Chapter Three: Larval Studies ........................................................................................ 24 Introduction........................................................................................................... 24 Materials and Methods.......................................................................................... 25 Collection and Maintenance of Bivalves.................................................. 25 Maintenance of Algal Cultures................................................................. 26 Preparation of Lysed Culture.................................................................... 26 Determination of Cell and Brevetoxin Concentrations............................. 27 Three-Day Static Exposure....................................................................... 27 Seven-Day Static Exposure ...................................................................... 29 Statistical Analysis.................................................................................... 30 Results................................................................................................................... 30 Toxin Profile of Karenia brevis Cultures ................................................. 30 Three-Day Static Exposure....................................................................... 31 Seven-Day Static Exposure ...................................................................... 32 Discussion............................................................................................................. 33 Chapter Four: Juvenile Studies....................................................................................... 49 Introduction........................................................................................................... 49 Materials and Methods.......................................................................................... 51 Collection and Maintenance of Juveniles ................................................. 51 Maintenance of Algal Cultures................................................................. 51 Preparation of Lysed Culture.....................................................................52 Determination of Cell and Brevetoxin Concentrations............................. 52 Clearance Rate Studies ............................................................................. 53 i Static Exposure Experiments ........................................................ 54 Flow-Through Exposure Experiments.......................................... 55 Results................................................................................................................... 57 Static Exposure Experiments .................................................................... 57 Bay scallops (Argopecten irradians) ............................................ 58 Green mussels (Perna viridis) ...................................................... 58 Northern quahogs (Mercenaria mercenaria)................................ 59 Eastern oysters (Crassostrea virginica)........................................ 59 Flow-Through Exposure Experiments...................................................... 60 Discussion............................................................................................................. 61 Chapter Five: Histopathology Studies ............................................................................ 81 Introduction........................................................................................................... 81 Materials and Methods.......................................................................................... 83 Results................................................................................................................... 85 Discussion............................................................................................................. 86 Chapter Six: General Discussion ..................................................................................... 95 Bivalve Larvae………………………………………………………………………97 Juvenile Bivalves…………………………………………………………………104 Conclusion and Significance………………………………………………………109 Impacts from Different Culture Preparations……………………………...109 Possible Mechanisms of Toxic Activity…………………………………...111 Implications for Fisheries Management…………………………………...114
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