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Developing Spawning Protocols and Embryological DEVELOPING SPAWNING PROTOCOLS AND EMBRYOLOGICAL BENCHMARKS FOR A TROPICAL MARINE FISH (Albula spp.) IN CAPTIVITY by William Robert Halstead III A Thesis Submitted to the Faculty of Charles E. Schmidt College of Science In Partial Fulfillment of the Requirements for the Degree of Master of Science Florida Atlantic University Boca Raton, FL December 2018 Copyright 2018 by William Robert Halstead III ii ACKNOWLEDGEMENTS I appreciate the guidance of my advisor, Dr. Paul Wills, and the support and input from my committee members, Dr. Aaron Adams, Dr. Matthew Ajemian, and Dr. Jon Shenker. I am grateful to everyone involved with the field and lab work required for this project including, at HBOI: Chris Robinson, Tony Cianciotto, Sahar Mejri, Richard Baptiste, Mike McCallister, Cameron Luck, and Alex Allen; and at CEI: Travis van Leeuwen, Anthony Cianciotto, Jake Rennert, Molly Wightman, Zack Nilles, Eric Schneider, Danielle Orrell, and several interns. Out of this list, I would like to highlight Anthony Cianciotto, Jake Rennert, and Molly Wightman for their singular efforts at CEI. Without the spawn they oversaw and the sleepless nights they endured monitoring and sampling the eggs and larvae, Chapter 2 of this thesis would not have been possible. Thank you to the Florida Fish and Wildlife Conservation Commission for approving the SAL to collect Bonefish in the Florida Keys and to Kathy Guindon of FFWCC Florida Wildlife Research Institute for doing species identification. At HBOI, I’m grateful for Dr. Annie Page-Karjian’s time and expertise for the ultrasound work. Dr. Katsumi Tsukamoto, Dr. Michael Miller, the students in their lab, and everyone at IRAGO were gracious hosts during my visit to Japan. Jimmy Nelson at HBOI and the staff of the boathouse at CEI were both very helpful and accommodating in the use their boats. iv This project was funded jointly by Bonefish & Tarpon Trust and the National Fish and Wildlife Foundation. This research was conducted in accordance with FAU IACUC protocols A16-34, A15-08, and A18-13. v ABSTRACT Author: William Robert Halstead III Title: Developing Spawning Protocols and Embryological Benchmarks for a Tropical Marine Fish (Albula spp.) in Captivity Institution: Florida Atlantic University Thesis Advisor: Dr. Paul Wills Degree: Master of Science Year: 2018 Relying on field research to complete the life history for certain fish species can be inadequate, but laboratory research can be used to fill these gaps. These gaps exist for Bonefish (Albula spp.), a tropical marine fish and popular sportfish. In this study, aquaculture techniques were applied to Bonefish in a captive setting at Harbor Branch Oceanographic Institute (HBOI) and Cape Eleuthera Institute (CEI) to induce spawning and describe early ontogeny. Photothermal manipulations and hormone injections were used to induce gonad maturation and spawning, which was achieved once at CEI and is the first record of hormone-induced spawning for Bonefish. From that spawn, egg and larval development were recorded and described through 26 hours and 56 hours respectively, representing the first record of these early life stages for Bonefish. vi This work expands upon what is known about Bonefish reproductive biology and will be useful for management and future captive research. vii DEDICATION I dedicate this thesis to my mother, Theresa, who instilled in me a lifelong love of learning, and my girlfriend, Colleen, whose patience and support during my project have meant everything. DEVELOPING SPAWNING PROTOCOLS AND EMBRYOLOGICAL BENCHMARKS FOR A TROPICAL MARINE FISH (Albula spp.) IN CAPTIVITY INTRODUCTION .................................................................................................. 1 CHAPTER 1 – CAPTIVE SPAWNING ................................................................ 14 Introduction ..................................................................................................... 14 Methods .......................................................................................................... 16 FAU-HBOI. ................................................................................................... 17 Collection. ................................................................................................. 17 Acclimation and husbandry. ...................................................................... 17 Gonad maturation and spawning. ............................................................. 18 Cape Eleuthera Institute (CEI). .................................................................... 22 Collection. ................................................................................................. 22 Acclimation and husbandry. ...................................................................... 23 Gonad maturation and spawning. ............................................................. 24 Results ............................................................................................................ 26 FAU-HBOI. ................................................................................................... 26 Collection. ................................................................................................. 26 Florida Keys. ......................................................................................... 26 Biscayne Bay. ........................................................................................ 27 Acclimation and husbandry. ...................................................................... 27 ix Florida Keys group. ............................................................................... 27 Biscayne group. ..................................................................................... 29 Gonad maturation and spawning. ............................................................. 29 Florida Keys group. ............................................................................... 29 Biscayne group. ..................................................................................... 35 CEI. .............................................................................................................. 37 Collection. ................................................................................................. 37 Long-term group. ................................................................................... 37 Short-term group. .................................................................................. 37 Pre-spawn aggregation group. .............................................................. 38 Acclimation and husbandry. ...................................................................... 38 Long-term group. ................................................................................... 38 Short-term group. .................................................................................. 39 Pre-spawn aggregation group. .............................................................. 39 Gonad maturation and spawning. ............................................................. 40 Long-term group. ................................................................................... 40 Short-term group. .................................................................................. 41 Pre-spawning aggregation group. ......................................................... 41 Discussion ....................................................................................................... 45 FAU-HBOI. ................................................................................................... 49 CEI. .............................................................................................................. 53 Conclusion ...................................................................................................... 57 CHAPTER 2 – EMBRYOLOGY AND LARVAL TIME SERIES ........................... 59 x Introduction ..................................................................................................... 59 Methods .......................................................................................................... 61 Spawning. .................................................................................................... 61 Incubation, hatching, and rearing. ................................................................ 62 Microscopy. .................................................................................................. 63 Classification. ............................................................................................... 64 Results ............................................................................................................ 65 Eggs. ............................................................................................................ 65 Larvae. ......................................................................................................... 68 Discussion ....................................................................................................... 69 Conclusion ...................................................................................................... 76 APPENDICES .................................................................................................... 78 Appendix A: Tables ......................................................................................... 79 Appendix B: Figures ...................................................................................... 100 REFERENCES ................................................................................................
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