54 JUNE 2023 • WORLD AQUACULTURE • WWW.WAS.ORG voluntarily, they will be strip-spawned by gentle application of pressure to the abdomen. No eggs were collected during the reporting period and only a small amount of non-viable sperm was collected. Therefore, in-vitro fertilization trials could not be performed or evaluated. Moving forward, the G2G project will continue broodstock maturation with the goal of collecting eggs during January-April 2023. Any voluntary egg production at that time will be recorded and reported, however, no hormone treatments will occur until a supply of cryopreserved sperm is available for in-vitro fertilization trials. The G2G project was very ambitious, especially considering that this project represents the first attempt to domesticate black grouper and red grouper for marine aquaculture. The concept of using visceral waste from commercial and recreational fishing as a source of valuable genetic material has potential application to other species and the application of cryopreservation for the purposes of in-vitro fertilization has the potential to transform aquaculture hatchery operations, as well as support conservation efforts for threatened and endangered species. Acknowledgements We thank Hunter Ledbetter from Hunter Sportfishing for providing many fresh grouper carcasses (both black grouper and red grouper), several female grouper broodstock and providing other fish samples for observation and practice cold storing gonads and sperm. We also want to thank Two Fish Charters for many initial fish sampled at the dock and the opportunity to participate in multiday fishing charters. We thank Key Largo Fisheries for contributing some of the largest black grouper sampled, Frank Levitsky for charter fishing in the Dry Tortugas, Captain Shane Leonard for charter fishing trips, and Lucas Mongelli for many grouper samples that always seemed to be female no matter how many times he tried. Special thanks to Tony Grogan and his preliminary research to inspire the PI to add to the research. Thanks to Murry Marine for storing pallets of fish food in their walk-in freezers. Thanks to the three research assistants ( Caeley Flowers, Piper Flowers and Jacob Harding), who took care of the black grouper broodstock and continue to work on this project. We thank the Florida Keys Commercial Fisherman’s Association for dissemination of information to the commercial fishers in the Florida Keys, but also to other grouper fisheries on the Florida West Coast. Finally, special thanks to the staff of US-1 Radio News, who showed continued interest in the project and allowed the PI to share monthly updates and spread awareness about the G2G project. Special thanks to Mr. Travis Knorr for initial plans and work on the G2G system and presenting information on the G2G system at WAS 2022. Thanks to Dr. James LaCosio and Dr. Chris Bennett for guidance and expertise on the design and equipment needed for the acoustic systems. Special thanks to Dr. Michelle Schärer for providing the recordings of black grouper spawning. This project received funding under [Subaward No. ACQ-210-039-2020CFK] from the Gulf States Marine Fisheries Commission in cooperation with NOAA Fisheries Service. The statements, findings, conclusions, and recommendations are those of the author(s) and do not necessarily reflect the views of Gulf States Marine Fisheries Commission or NOAA Fisheries. Notes Patrick H. Rice and Caeley V. Flowers, College of the Florida Keys, 5901 College Rd., Key West, FL 33040 Principal Investigator: Patrick H. Rice, College of the Florida Keys, 5901 College Rd., Key West, FL 33040 Co-Principal Investigator: David E. Vaughan, Dr. David E. Vaughan Consulting, 24215 Caribbean Dr. W., Summerland Key, FL 33042 1 104.1 FM: US-1 Radio; 101.7 FM: Pirate Radio; 102.5 FM: Keys Talk Radio 2 US-1 Radio: This Week in Fishing 3 US-1 Radio: Morning Magazine and Evening Edition 4 101.7 FM: Pirate Radio; 102.5 FM: Keys Talk Radio 5 Leica Model CME, Focus USA Technologies, Inc. 6 U.S. Solid Cryogenics, Model: USS-LNT00003 7 FishFreeze Cryopreservation Milt Freezing Media, Syndel Inc. 8 CARSC, Model Hei012, amazon.com 9 Dr. Doug Mader, personal communication 10 Madmac-MS maturation stimulator and broodstock conditioner; Aquafauna Bio-Marine, Inc. References Amagai, T., D. Izumida, R. Murata and K. Soyano. 2022. Male pheromones induce ovulation in female honeycomb groupers (Epinephelus merra): a comprehensive study of spawning aggregation behavior and ovarian development. Cells 11(3):484. Blaxter, J.H.S. 1953. Sperm storage and cross-fertilization of spring and autumn spawning herring. Nature 172:1189-1190. Gwo, J.C. 1993. Cryopreservation of black grouper (Epinephelus malabaricus) spermatozoa. Theriogenology 39:1331-1342. Gwo, J.C., B.G.M. Jamieson and L.K.P. Leung. 2009. Live preservation of fish gametes. Pages 395-483 In: Chapter. 11, Reproductive Biology and Phylogeny of Fishes (Agnathans and Bony Fishes), B.G.M Jamieson, editor. CRC Press, Boca Raton, FL. Marino, G., E. Panini, A. Longobardi, A. Mandich, M.G. Finoia, Y. Zohar and C.C. Mylonas. 2003. Induction of ovulation in captivereared dusky grouper, Epinephelus marginatus (Lowe, 1834), with a FIGURE 4. Photo of the G2G monitoring system showing images from each tank with the habitat structure. Notice two black grouper using the habitat in Tank 2 (Photo: Patrick Rice).
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