Conservation of Freshwater Live-Bearing Fishes: Development
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Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 7-6-2018 Conservation of Freshwater Live-bearing Fishes: Development of Germplasm Repositories for Goodeids Yue Liu Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Aquaculture and Fisheries Commons, Biotechnology Commons, and the Cell Biology Commons Recommended Citation Liu, Yue, "Conservation of Freshwater Live-bearing Fishes: Development of Germplasm Repositories for Goodeids" (2018). LSU Doctoral Dissertations. 4675. https://digitalcommons.lsu.edu/gradschool_dissertations/4675 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. CONSERVATION OF FRESHWATER LIVE-BEARING FISHES: DEVELOPMENT OF GERMPLASM REPOSITORIES FOR GOODEIDS A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The School of Renewable Natural Resources by Yue Liu B.S., Jiujiang University, 2010 M.Agric., Shanghai Ocean University, 2013 August 2018 For my maternal grandparents, Wenzhi Zhang and Xianrang Zhang, who raised me up in my childhood For my parents, who support me with all their love For Youjin and Jenna, who are the meaning of my life ii Acknowledgments I want to thank my advisor Dr. Terrence Tiersch, who has been the most important person in my PhD study. I still vividly remember the great encouragement he gave to me when the first time I met him with my awkward “Chinglish”. Since that, he has encouraged me to become a good scientist, and showed me how to learn, think, do research, write, teach, communicate with groups, manage laboratories, collaborate, work hard, persist, and deal with opportunities and difficulties. I thank my PhD advisory committee for their valuable advice on this dissertation. I thank Dr. Henrique Cheng for accepting me into his laboratory, teaching me research techniques, and giving me chances to participate in academic activities within the LSU School of Veterinary Medicine. I thank Dr. Maria Teresa Gutierrez-Wing for always giving me advice on research, opening my mind to the engineering world, and encouraging me to learn not only from thinking, but by trials and failures. I thank Dr. Wei Xu for providing valuable insights on genomic and molecular biology. I thank Dr. Sabrina Taylor for teaching me knowledge and methods of conservation genetics. I thank Dr. James Cowan for serving as Dean’s representative. I am very thankful to all of the people at the Aquatic Germplasm and Genetic Resources Center. I thank William Childress, Amy Guitreau, and Michael Christensen for their friendship and tremendous help in my research and daily life. I thank Dr. Leticia Torres for sharing me her experiences on research, career development and parenting. I thank Dr. Huiping Yang for training me in laboratory skills and leading me into the world of live-bearing goodeids. I thank Dr. E Hu and Dr. Noel Novelo for taking care of me like big brothers at the beginning of my student life in the United States. I thank undergraduate student workers: Jacob Schaff, Jack Cane, Henry Salassi, David Thomas, Patrick Cole, Tommy Lee, Kyleigh Ardoin, Sarah Thomas, Loan iii Phan, Leonard Bourgeois, and any others whose names are not listed here. I thank former students from our laboratory who I have encountered for their kind encouragement and guidance: Dr. Rafael Cuevas-Uribe, Dr. John Buchanan, Dr. Carmen Paniagua-Chavez, and Dr. William Wayman. I thank other scientists and researchers that have contributed greatly to this dissertation. I especially thank Dr. Harry Grier for his inspiration, suggestions, and for providing fish. I thank Dr. John Hawke of the LSU School of Veterinary Medicine for diagnosis and advice on fish diseases. I thank Dr. Sokolova Yuliya for assistance with processing and reviewing samples for electronic scanning microscopy. I thank Kendra Shultz for processing samples for histological study. I also thank Dr. John Lyons, Dr. Kyle Piller, Dr. Mari Carmen-Uribe, and Dr. Omar Domínguez-Domínguez for providing encouragement and insights on conservation of imperiled goodeids. I am grateful for my beloved family. I thank my parents for their love and support throughout my life. They have supported me whenever I needed it, taught me how to be a man, and encouraged me to help the world. I thank my wife for encouraging me and taking care of me and our family. I thank my daughter who gives me energy whenever I am feeling challenged. I thank my maternal grandparents who took great care of me in my childhood and always thought of me. I thank all my other relatives in Qinhuangdao, Korea, Changchun, and Beijing, who encourage me all of the time and take care of my family unconditionally. iv Table of Contents Acknowledgments.......................................................................................................................... iii List of Tables ................................................................................................................................ vii List of Figures .............................................................................................................................. viii Abstract ........................................................................................................................................ xiii Chapter 1 Introduction ..................................................................................................................................... 1 References .............................................................................................................................. 14 Chapter 2 Background ................................................................................................................................... 17 The Conservation Crisis of Live-bearing Goodeids .............................................................. 17 Conservation Tools and Limitations ...................................................................................... 19 Germplasm Repositories are a Necessary Conservation Tool ............................................... 22 Importance of Live-bearing Fishes ........................................................................................ 23 Unusual Reproductive Characteristics of Live-bearing Fishes .............................................. 24 Reproducibility and Standardization ..................................................................................... 25 Sperm Bundles ....................................................................................................................... 27 Motility Activation ................................................................................................................ 29 Signaling Mechanisms of Sperm Activation ......................................................................... 31 Development of Cryopreservation Protocols ......................................................................... 31 Internal Fertilization .............................................................................................................. 33 Summary ................................................................................................................................ 35 References .............................................................................................................................. 35 Chapter 3 Quality Evaluation of Sperm from Live-bearing Fishes: Standardized Assessment of Sperm Bundles from Redtail Splitfin (Xenotoca eiseni, Rutter, 1896) .................................................... 43 Materials and Methods ........................................................................................................... 46 Results .................................................................................................................................... 53 Discussion .............................................................................................................................. 56 Summary ................................................................................................................................ 60 References .............................................................................................................................. 60 Chapter 4 Activation of Free Sperm and Dissociation of Sperm Bundles from Redtail Splitfin .................. 65 Materials and Methods ........................................................................................................... 69 Results .................................................................................................................................... 77 Discussion .............................................................................................................................. 87 Summary ................................................................................................................................ 94 References .............................................................................................................................