Effects of cryopreservation on the survival rate of the seven-band grouper (Epinephelus septemfasciatus) embryos |
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| Institution: | 1. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;3. Ming Bo Aquatic Co. Ltd., Laizhou 261400, China;1. Department of Aquaculture, Szent István University, Páter Károly u. 1, H-2100 Gödöll?, Hungary;2. Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 2, 21000 Novi Sad, Serbia;3. Department of Animal Science, Biotechnical Faculty, University of Ljubljana, Groblje 3, Sl-1230 Dom?ale, Slovenia;1. Department of Neuroscience and Regenerative Medicine, Georgia Regents University, Augusta, GA 30912, USA;2. Institute of Biomedical Science and Environmental Science and Technology, University of Bedfordshire, Luton, Bedfordshire LU2 8DL, UK;3. School of Applied Science, Bournemouth University, Poole BH12 5BB, UK;1. Grupo de Acuicultura y Biodiversidad, Instituto de Ciencia y Tecnología Animal, Universitat Politècnica de València, Camino de Vera s/n 46022, Valencia, Spain;2. CCMAR, University of Algarve, Campus of Gambelas, 8005-139 Faro, Portugal;3. Department of Aquaculture, Szent István University, 2100 Gödöll?, Páter Károly u. 1., Hungary;1. Centre of Marine Sciences-CCMAR, University of Algarve, 8005-139, Faro, Portugal;2. Portuguese Institute for Sea and Atmosphere-IPMA, Av. 5 de Outubro, 8700-305, Olhão, Portugal;3. Necton, S.A., Belamandil, 8700-152, Olhão, Portugal |
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| Abstract: | The effects of cryopreservation and the vitrification solution on the embryo hatchability of the seven-band grouper Epinephelus septemfasciatus were evaluated in this study. Six small molecule cryoprotectants (PG, MeOH, Gly, DMF, DMSO and EG) and four macromolecular cryoprotectants (glucose, fructose, sucrose and trehalose) were used to determine the embryo toxicity levels. Results showed that the embryo survival rate was higher when the PM (24% PG + 16% MeOH):Gly ratios were 3:1 and 4:1. Further experiments showed that the embryo survival rates in PMG3S (35% PMG3 + 5% sucrose) and PMG3T (35% PMG3 + 5% trehalose) were relatively higher, which are 29.24 ± 10.81% and 27.01 ± 3.39%, respectively. When treated with PMG3S and PMG3T by using 5-step method, embryos at somite stage and tail-bud stage shrank in the first 6 min and gradually recovered in volume to the original. This indicated the successful permeation of the vitrification solutions into cells. Then, embryos at the embryoid body formation stage, the somite stage and the tail-bud stage were cryopreserved with PMG3S and PMG3T. In total, 82 floating embryos were obtained, 14 of which developed further, with 8 embryos at the tail-bud stage developing to the heartbeat stage, 4 embryos at the body formation stage development to the somite stage, and 2 embryos at the somite stage hatched to larval fish. |
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| Keywords: | Cryopreservation Vitrification Embryos Hatchability |
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