ASSESSMENT OF CULTURE TECHNIQUES FOR TWO Halichoeres WRASSES, H. melanurus AND H. chrysus By ELIZABETH MARIE GROOVER A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2018 © 2018 Elizabeth Marie Groover To all who have supported me in pursuit of my passion ACKNOWLEDGMENTS I would like to thank my friends and family, especially my parents, Michael and Donna Groover, whose unconditional love and support have made me the woman I am today. I would like to acknowledge my major professor, Dr. Matthew DiMaggio, for his experience, guidance, encouragement, and confidence in my abilities as a scientist and fish culturist. I would also like to acknowledge my committee members, Dr. Cortney Ohs and Dr. Joshua Patterson, for their assistance and expertise in the field. Thank you to my fellow colleagues Dr. Katharine Starzel, Michael Sipos, Tim Lyons, Taylor Lipscomb, Dr. Marion Hauville, and Shane Ramee for their advice, assistance, knowledge, and friendship. Additionally, I would like to thank all staff and students at University of Florida’s Tropical Aquaculture Lab for their support, knowledge, guidance, and willingness to help whenever it was needed. I would like to thank Segrest Farms and Kevin Kohen of Doctors Foster and Smith Live Aquaria.com for their generous donations of wrasse broodstock, as well as Instant Ocean Spectrum Brands, Larry’s Reef Services, Fritz Industries, Inc., Piscine Energetics, and Petco for providing additional elements needed for these trials. Lastly, I would like to thank Dr. Judy St. Leger, Rising Tide Conservation, and the Sea World and Busch Gardens Conservation Fund for providing the funding for my research. TABLE OF CONTENTS page ACKNOWLEDGMENTS .................................................................................................. 4 LIST OF TABLES ............................................................................................................ 8 LIST OF FIGURES .......................................................................................................... 9 ABSTRACT ................................................................................................................... 11 CHAPTER 1 GENERAL INTRODUCTION .................................................................................. 13 Marine Aquarium Trade .......................................................................................... 13 Marine Ornamental Aquaculture ............................................................................. 14 Labridae .................................................................................................................. 17 Halichoeres ............................................................................................................. 19 2 THE MELANURUS WRASSE Halichoeres melanurus: A DESCRIPTION OF SPAWNING, CULTURE PROTOCOLS, LARVAL DEVELOPMENT, AND COMPLETION OF LIFE CYCLE ............................................................................. 21 Introduction ............................................................................................................. 21 Methods .................................................................................................................. 24 Acquisition of Broodstock ................................................................................. 24 Quarantine Protocol ......................................................................................... 24 Broodstock Husbandry ..................................................................................... 26 Egg Collection and Analysis ............................................................................. 27 Larval Growth and Development Trial .............................................................. 27 Results .................................................................................................................... 31 Quarantine Protocol ......................................................................................... 31 Spawning Frequency and Quantity .................................................................. 31 Larval Growth and Development Trial .............................................................. 31 Completion of Life Cycle................................................................................... 35 Discussion .............................................................................................................. 36 3 DEVELOPMENT OF EARLY LARVICULTURE PROTOCOLS FOR Halichoeres melanurus ............................................................................................................... 50 Introduction ............................................................................................................. 50 Materials and Methods............................................................................................ 56 Acquisition and Quarantine of Broodstock........................................................ 56 Broodstock Husbandry ..................................................................................... 58 Egg Collection and Analysis ............................................................................. 59 Experimental Design of Embryo Incubation Experiments ................................. 60 5 Experiment 3-1 Temperature - Embryo Incubation Time and Larval Survival at Hatch ......................................................................................................... 61 Experiment 3-2 Temperature - Larval Size at Hatch ........................................ 61 Experimental Design of Larviculture Experiments ............................................ 62 Experiment 3-3 Algal Density - Larval Growth and Survival ............................. 64 Experiment 3-4 Shading Method - Larval Growth and Survival ........................ 64 Experiment 3-5 Algal Density and Prey Availability Period - Larval Feeding Percentage .................................................................................................... 65 Experiment 3-6 First Feed Type and Prey Availability Period - Larval Feeding Percentage ...................................................................................... 66 Experiment 3-7 Prey Density and Prey Availability Period - Larval Feeding Percentage .................................................................................................... 66 Statistical Analysis ............................................................................................ 67 Results .................................................................................................................... 68 Experiment 3-1 and 3-2 Temperature - Incubation Time, Larval Size and Survival ......................................................................................................... 68 Experiment 3-3 Algal Density - Larval Growth and Survival ............................. 69 Experiment 3-4 Shading Method - Larval Growth and Survival ........................ 69 Experiment 3-5 Algal Density and Feed Availability Period - Larval Feeding Percentage .................................................................................................... 70 Experiment 3-6 First Feed Type and Feed Availability Period - Larval Feeding Percentage ...................................................................................... 70 Experiment 3-7 Prey Density and Feed Availability Period - Larval Feeding Percentage .................................................................................................... 70 Discussion .............................................................................................................. 71 4 EXPLORATION OF EARLY LARVICULTURE PROTOCOLS FOR Halichoeres chrysus ................................................................................................................... 83 Introduction ............................................................................................................. 83 Materials and Methods............................................................................................ 89 Acquisition and Quarantine of Broodstock........................................................ 89 Broodstock Husbandry ..................................................................................... 91 Egg Collection and Analysis ............................................................................. 92 Experimental Design of Embryo Incubation Experiments ................................. 93 Experiment 4-1 Temperature - Embryo Incubation Time and Larval Survival at Hatch ......................................................................................................... 94 Experiment 4-2 Temperature - Larval Size at Hatch ........................................ 94 Experimental Design of Larviculture Experiments ............................................ 95 Experiment 4-3 Algal Density - Larval Growth and Survival ............................. 97 Experiment 4-4 Shading Method - Larval Growth and Survival ........................ 97 Experiment 4-5 Algal Density and Prey Availability Period – Larval Feeding Percentage .................................................................................................... 98 Experiment 4-6 Prey Density and Prey Availability Period - Larval Feeding Percentage ...................................................................................................