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Optimization of vitrification factors for embryo cryopreservation of kelp grouper (Epinephelus moara)

文献类型： 外文期刊

作者： Zhang, Jingjing 1 ; Tian, Yongsheng 1 ; Li, Zhentong 1 ; Wu, Yuping 1 ; Li, Ziqi 1 ; Cheng, Meiling 1 ; Wang, Linna 1 ; M 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China

2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266000, Shandong, Peoples R China

3.Shanghai Ocean Univ, Natl Demonstrat Ctr Expt Fisheries Sci Educ, Shanghai 201306, Peoples R China

4.Dalian Ocean Univ, Coll Fisheries & Life Sci, Dalian 116023, Peoples R China

5.Ming Bo Aquat Co Ltd, Laizhou 261400, Peoples R China

关键词： Cryopreservation; Embryo; Hatchability; Abnormality; Epinephelus moara

期刊名称：THERIOGENOLOGY （ 影响因子：2.74； 五年影响因子：2.93 ）

ISSN： 0093-691X

年卷期： 2020 年 142 卷

页码：

收录情况： SCI

摘要： Cryopreservation of marine fish embryos causes to severe cryogenic damage, and to date, adults have not been reared from embryos that were cryopreserved. Here, we optimized vitrification factors to improve the survival and hatching rate of kelp grouper (Epinephelus moara) embryos after cryopreservation. We screened the effects of 11 vitrification solution concentrations (25-50%) on the survival rate of embryos at four developmental stages (16S, 185, 225, TB). We investigated the effects of different equilibration time (25-45min) on the survival rate and the influence of vitrification solutions on embryonic volume. In addition, we tested the effects of treating embryos at five different developmental stages (4-6S, 16S, 22S, TB, HB) with different vitrification solutions (35% PMG3S and 35% PMG3T), prechilling temperature (-5 degrees C and 4 degrees C) and prechilling time. In total, 9855 embryos were cryopreserved at 10 developmental stages, from optic capsule stage to pre-hatch stage. We found that kelp grouper embryos performed best at equilibration time of 30 min. Embryos at the tail-bud stage exhibited greater tolerance to vitrification than other stages. Vitrification solutions that contained sucrose showed better survival rates compared to embryos treated with vitrification solutions containing trehalose. Pre-chilling treatment improved viability before freezing, but did not improve viability after freezing. In the most optimal condition we identified in this study, the average survival, normal development and malformation rates of cryopreserved embryos were 6.32%, 2.36% and 3.49%, and 39.85% of the surviving embryos that were cryopreserved hatched. The hatched larvae gradually died at day 12 of cultivation, where the longest surviving individuals lived for 16 days. This study provides valuable data for improving survival and hatching rate of cryopreserved grouper embryos, and provides references for further exploring techniques in fish embryo cryopreservation. (C) 2019 Elsevier Inc. All rights reserved.

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