Developing Embryo Technologies for the Eland Antelope (Taurotragus Oryx) Gemechu G
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Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2004 Developing embryo technologies for the eland antelope (Taurotragus oryx) Gemechu G. Wirtu 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 Veterinary Medicine Commons Recommended Citation Wirtu, Gemechu G., "Developing embryo technologies for the eland antelope (Taurotragus oryx)" (2004). LSU Doctoral Dissertations. 505. https://digitalcommons.lsu.edu/gradschool_dissertations/505 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]. DEVELOPING EMBRYO TECHNOLOGIES FOR THE ELAND ANTELOPE (TAUROTRAGUS ORYX) 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 Interdepartmental Program in Veterinary Medical Sciences through the Department of Comparative Biomedical Sciences by Gemechu Wirtu D.V.M., Addis Ababa University, 1992 M.S., Virginia Polytechnic Institute and State University, 1999 May 2004 Acknowledgments My dream to pursue graduate studies in the USA became a reality when my application for the Fulbright Scholarship was miraculously approved in 1997. I am deeply indebted to all involved at that critical stage. At Louisiana State University (LSU) and Audubon Center for Research of Endangered Species (ACRES), I am grateful to those who were helpful in my getting admission to LSU graduate school. I am grateful to my graduate Committee Members: Drs. Charles Short, Earle Pope and Robert Godke (Committee Co-chairs) and Drs. Barry Bavister, Leonard Kappel and Changaram Venugopal (Committee members). I also thank Dr. James Cronin for valuable comments on my dissertation. Drs. Pope, Bavister and Godke closely supervised aspects of my research. I am deeply indebted to Dr. Betsy Dresser, Director of ACRES, for continued support and confidence in me. Many people at ACRES had important inputs in my project. From the Veterinary Research Unit, Dr. Susan Mikota (made me feel at home and had valuable inputs at the inception of the project idea); Dr. Chris Janssen assisted with designing custom-made adaptor for the ultrasound transducer; Barbara Vincent and Contessa Knight were always ready to offer their help to me. Dr. Alexander Cole attended most of the eland procedures and I am thankful for his full collaboration. I also had valuable discussions with Dr. Ellen Boyd, a fellow graduate student. Drs. Roberto Aguilar and Steve Miller of the Audubon Institute also attended some of the oocyte retrieval procedures. From Species Survival Center, I am especially grateful to Jeff Vaccaro and Erin Sarrat who have been cooperative in all aspects of my project that required their involvement. I could not have done the eland handling part of my project without their creative inputs and facilitative roles. Thank you very much. I should also mention the valuable support I received from Carol Allen, Lola Curtis, Dierdre Havnen and many interns and volunteers who assisted me over the last four years. I received invaluable assistance from personnel from Dr. Pope and Dr. Bavister laboratories. From Dr. Pope’s laboratory I would like to thank Amy King, Rebecca Harris, Dr. Martha Gomez, ii Lola Curtis, Angelica Giraldo, Cherie Dumas, Kia Gray and Liesl Nel-Themaat; and from Dr. Bavister’s laboratory, Kimberly Poole, Dr. Jane Andrews, Dr. Byeongchun Lee, Inger Carlson, Denise Kinsey and Stephanie Nichols. Dr. Philip Damiani also played crucial roles by doing the nuclear transfer and intracytoplasmic sperm injection. He also was responsible for acquiring most of the bovine oocytes that were used for the bovine embryo culture experiments. Gwen Alleman, Celestine Washington, Josie Wolfe, Alicia Benoit, Stella Sullivan and Jackie Coulon of ACRES are acknowledged for administrative and moral support. I also thank Dr. Stanley Leibo of ACRES/University of New Orleans for allowing me access to his large collection of reference materials. From the Department of Comparative Biomedical Sciences (previously Veterinary Physiology, Pharmacology and Toxicology), I thank Dr. Steven Barker, Coordinator of Graduate Programs, and Mona Busby, Shelia Rigby and Jeanie Tyler for their administrative support. I am also grateful to Dr. Ramaswamy for the friendship and helping me in many ways. Similarly, Dr. Asfaw Bekele of Tarleton State University helped me with statistics and by sending many articles online. Dr. Tadele Kiros of the University of Saskatchewan (Canada) also offered a similar assistance. The work presented here was partly supported by grants from Johnston Science Foundation and the Coypu foundation. Many more people had extended their full support to me and the absence of your names from list does not mean I am not grateful…Thank you all. Finally, to my family and children: thank you for your patience and understanding during my prolonged disappearances from your vicinity. In addition, I will always remain grateful to my parents, who took the initial step of sending me to school by breaking the tradition of their parents and community. iii Table of Contents ACKNOWLEDGMENTS.....................................................................................................................ii LIST OF TABLES ................................................................................................................................v LIST OF FIGURES............................................................................................................................vii ABSTRACT.......................................................................................................................................viii CHAPTER 1. INTRODUCTION ..........................................................................................................1 CHAPTER 2. LITERATURE REVIEW................................................................................................4 2.1. THE ROLE OF REPRODUCTIVE TECHNOLOGIES IN BIODIVERSITY CONSERVATION .........................4 2.2. GAMETE RETRIEVAL AND IN VITRO EMBRYO PRODUCTION IN FARM AND NONDOMESTIC MAMMALS ...........................................................................................................................................6 2.3. ASSISTED REPRODUCTIVE TECHNOLOGIES IN THE COMMON ELAND ...........................................18 CHAPTER 3. DEVELOPMENT OF IN VITRO-DERIVED BOVINE EMBRYOS IN DEFINED (PROTEIN-FREE) MEDIA .................................................................................................................38 3.1. INTRODUCTION ...........................................................................................................................38 3.2. MATERIAL AND METHODS ..........................................................................................................39 3.3. RESULTS .....................................................................................................................................45 3.4. DISCUSSION ................................................................................................................................53 CHAPTER 4. HANDLING ELANDS FOR REPRODUCTIVE TECHNOLOGY PROCEDURES WITHOUT INDUCING GENERAL ANESTHESIA: ROLES OF BEHAVIORAL TRAINING AND HYDRAULIC CHUTE ..............................................................................................................60 4.1. INTRODUCTION ...........................................................................................................................60 4.2. MATERIALS AND METHODS ........................................................................................................62 4.3. RESULTS .....................................................................................................................................69 4.4. DISCUSSION ................................................................................................................................80 CHAPTER 5. TRANSVAGINAL ULTRASOUND-GUIDED OOCYTE COLLECTION, IN VITRO OOCYTE MATURATION AND EMBRYO PRODUCTION IN THE ELAND ANTELOPE .........................................................................................................................................88 5.1. INTRODUCTION ...........................................................................................................................88 5.2. MATERIALS AND METHODS ........................................................................................................90 5.3. RESULTS ...................................................................................................................................103 5.4. DISCUSSION ..............................................................................................................................115 CHAPTER 6. SUMMARY AND CONCLUSIONS .........................................................................122 REFERENCES...................................................................................................................................126 APPENDIX A: ATTRIBUTES OF SPECIES OF TRAGELAPHINE ANTELOPES...............................................................155