title: Effects of cryopreservation at various temperatures on the survival of kelp grouper (Epinephelus moara) embryos from fertilization with cryopreserved sperm.
abstract: Fish embryo cryopreservation is highly important for the long-term preservation of genomic and genetic information; however, few successful cases of fish embryo cryopreservation have been reported over the past 60 years. This is the first study to use Epinephelus moara embryos from fertilization with cryopreserved sperm as experimental material. Embryos that developed to the 16-22 somite stage and tail-bud stage were treated with the vitrification solution PMG3T according to a five-step equilibration method and cryopreserved at various temperatures and storage duration. Only 19.9 ± 9.2% of 16-22 somite stage embryos and 1.3 ± 1.1% of tail-bud stage embryos survived when cooled at 4 °C for 60 min. In total, 8.0 ± 3.0% of 16-22 somite stage embryos survived when cooled at -25.7 °C for 30 min, 22.4 ± 4.7% of tail-bud stage embryos survived after 45 min of cooling at -25.7 °C, and none survived after 60 min. Only 2.0 ± 2.7% of embryos survived when cryopreserved at -140 °C for 20 min. However, 9.7% of tail-bud stage embryos survived after cryopreservation in liquid nitrogen (-196 °C) for 2 h. Most surviving embryos developed normally. Embryonic volume decreased and spherical segments appeared when embryos were treated with higher concentrations of vitrification solution. Additionally, the volume recovered gradually after rinsing with sucrose and seawater. This is the first estimate of the survival of E. moara embryos and larvae after cryopreservation. These findings provide a foundation for further explorations of fish embryo cryopreservation techniques.
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266000, China. Electronic address: tianys@ysfri.ac.cn.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266000, China.
authors.forename: Zhangfan
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266000, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Tang
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266000, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.
authors.forename: Huimin
authors.initials: H
authors.lastname: Duan
authors.affiliation: Ming Bo Aquatic Co. Ltd., Laizhou 261400, China.
authors.forename: Jieming
authors.initials: J
authors.lastname: Zhai
authors.affiliation: Ming Bo Aquatic Co. Ltd., Laizhou 261400, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Li
authors.affiliation: Ming Bo Aquatic Co. Ltd., Laizhou 261400, China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Ma
authors.affiliation: Ming Bo Aquatic Co. Ltd., Laizhou 261400, China.
authors.forename: Jiangchun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Ming Bo Aquatic Co. Ltd., Laizhou 261400, China.
authors.forename: Yunxia
authors.initials: Y
authors.lastname: Hou
authors.affiliation: Ming Bo Aquatic Co. Ltd., Laizhou 261400, China.
authors.forename: Zhengxiang
authors.initials: Z
authors.lastname: Sun
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods* / Q000662:veterinary; D004625:Embryo, Nonmammalian; D005260:Female; D005306:Fertilization; D008297:Male; D010473:Perciformes; D013094:Spermatozoa; D013696:Temperature; D058989:Vitrification
keywords: Embryo cryopreservation; Epinephelus moara; Frozen sperm