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Fundulus Heteroclitus) Reproduction Under Different Salinities Chelsea O'connor Blatchley University of Connecticut - Storrs, [email protected]

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Fundulus Heteroclitus) Reproduction Under Different Salinities Chelsea O'connor Blatchley University of Connecticut - Storrs, Chelsea.Blatchley@Uconn.Edu University of Connecticut OpenCommons@UConn Master's Theses University of Connecticut Graduate School 7-19-2013 The ffecE ts of a Model Androgen 5a- dihydrotestosterone on Mummichog (Fundulus heteroclitus) Reproduction Under Different Salinities Chelsea O'Connor Blatchley University of Connecticut - Storrs, [email protected] Recommended Citation Blatchley, Chelsea O'Connor, "The Effects of a Model Androgen 5a-dihydrotestosterone on Mummichog (Fundulus heteroclitus) Reproduction Under Different Salinities" (2013). Master's Theses. 469. https://opencommons.uconn.edu/gs_theses/469 This work is brought to you for free and open access by the University of Connecticut Graduate School at OpenCommons@UConn. It has been accepted for inclusion in Master's Theses by an authorized administrator of OpenCommons@UConn. For more information, please contact [email protected]. The Effects of a Model Androgen 5α-dihydrotetsosterone on Mummichog (Fundulus heteroclitus) Reproduction Under Different Salinities Chelsea O’Connor Blatchley B.S., University of Rhode Island, 2010 A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science At the University of Connecticut 2013 ii Acknowledgments I would like to thank all of the faculty, staff, and students at the University of Connecticut who have helped me complete this thesis project. It has truly been a collaborative effort and I would not have been able to do this project without them. Thank you especially to my advisor Dr. Bosker who has helped to guide me through this process by offering his invaluable expertise in the field of environmental toxicology. Thank you also to each of my committee members, Dr. DeGuise, Dr. Vokoun, Dr. Frasca and Dr. Provatas, for asking tough questions and providing me with their assistance and expertise throughout this whole process. Thank you also to the folks at CESE who have kindly offered their assistance and instruction: Gary Ulatowski, Chris Perkins, Aliaksander Yeudakimau, Phil Caron, and Paul Fogle. Thank you to the undergraduate lab technicians, Christine Taylor and Steve Cranford, who spent many long hours on this project over the summer. Thanks also to Dr. Levin, Kelly O’Connor, and Jordan Rabon for helping out on our big dissection days. A special thanks to Tanya Lama for her hard work, dedication, and companionship throughout the project. Lastly, I would like to thank my parents, my brother, Tyler, my friends, and my fiancé Dave for their love, support, and encouragement along the way. Funding for this project was provided by the University of Connecticut. iii Table of Contents General Abstract ............................................................................................................................ vi Chapter 1 General Introduction ................................................................................................. 1 1.1 Endocrine Disrupting Substances (EDSs) ........................................................................ 2 1.2 Sources of EDSs ............................................................................................................... 4 1.3 Androgenic EDSs ............................................................................................................. 5 1.4 The influence of salinity on fish response to EDSs ......................................................... 7 1.5 Standard 21-day reproductive bioassay............................................................................ 9 1.6 Modeling population level effects from EDS exposure ................................................. 10 1.7 Mummichog as a toxicological model ........................................................................... 11 1.8 Objectives and Hypotheses ............................................................................................ 12 1.9 References ...................................................................................................................... 15 Chapter 2 The impact of model androgen 5α-dihyrdotestosterone on mummichog (Fundulus heteroclitus) under different salinities .......................................................................................... 23 2.1 Abstract .......................................................................................................................... 24 2.2 Introduction .................................................................................................................... 26 2.3 Methods .......................................................................................................................... 31 2.3.1 Chemical ................................................................................................................. 31 2.3.2 Fish .......................................................................................................................... 31 2.3.3 Short-term reproductive bioassay ........................................................................... 32 2.3.4 Fish sampling and reproductive endpoints ............................................................. 34 2.3.5 ELISA for in vitro sex steroid production .............................................................. 35 2.3.6 Histological analysis ............................................................................................... 35 2.3.7 Solid phase extraction water chemistry analysis .................................................... 38 2.3.8 Statistics .................................................................................................................. 40 2.4 Results ............................................................................................................................ 43 2.4.1 Somatic endpoints ................................................................................................... 43 2.4.2 In vitro hormone levels ........................................................................................... 47 2.4.3 Histology ................................................................................................................. 50 2.4.4 Gonadal somatic index ............................................................................................ 60 2.4.5 Egg production ........................................................................................................ 60 iv 2.4.6 Water chemistry ...................................................................................................... 66 2.5 Discussion ...................................................................................................................... 68 2.5.1 The influence of salinity on DHT treatment ........................................................... 68 2.5.2 Effects of DHT on mummichog reproduction ........................................................ 76 Appendix A ................................................................................................................................... 84 2.6 References ...................................................................................................................... 87 Chapter 3 Modeling population level effects of mummichog (Fundulus heteroclitus) exposed to 5α-dihydrotestosterone ........................................................................................................... 100 3.1 Abstract ........................................................................................................................ 101 3.2 Introduction .................................................................................................................. 102 3.3 Methods ........................................................................................................................ 107 3.3.1 Development of a mummichog population model................................................ 107 3.3.2 Model application ................................................................................................. 107 3.3.3 Effects of exposure to stressors on population status ........................................... 112 3.3.4 Subspecies differences .......................................................................................... 114 3.4 Results .......................................................................................................................... 118 3.5 Discussion .................................................................................................................... 124 Appendix A ................................................................................................................................. 130 3.6 References .................................................................................................................... 134 Chapter 4 General discussion ................................................................................................ 142 4.1 References .................................................................................................................... 146 v General Abstract Endocrine disrupting substances (EDSs) have the potential to disrupt sensitive hormone pathways, including those involved in development and reproduction. Both fresh and estuarine water bodies receive inputs of EDSs from a variety of sources, including sewage effluent, industrial effluent, and agricultural runoff. Based on limited available literature, freshwater species appear to respond to lower levels of EDSs than estuarine or marine species. To address this
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