Cryopreservation of embryos of humpback grouper (Cromileptes altivelis) using combinations of non-permeating cryoprotectants

文献类型： 外文期刊

第一作者： Li, Linlin

作者： Li, Linlin;Tian, Yongsheng;Li, Zhentong;Li, Ziqi;Duan, Pengfei;Wang, Xinyi;Chen, Shuai;Wang, Linna;Li, Linlin;Tian, Yongsheng;Wang, Linna;Li, Zhentong;Li, Ziqi;Duan, Pengfei;Wang, Xinyi;Chen, Shuai;Wang, Qingbin;Zhai, Jieming

作者机构：

关键词： Combined cryoprotectant; Cryopreservation; Grouper embryo; Non-permeating cryoprotectant

期刊名称：AQUACULTURE （ 影响因子：5.135； 五年影响因子：5.125 ）

ISSN： 0044-8486

年卷期： 2022 年 548 卷

页码：

收录情况： SCI

摘要： In this study, Cromileptes altivelis embryos were successfully preserved using non-permeable cryoprotectants for the first time. We compared the toxicity of single non-permeable cryoprotectants and combination of nonpermeable cryoprotectants on C. altivelis embryos. Five non-permeable cryoprotectants at 1 M (i.e., trehalose, sucrose, lactose, fructose, and glucose) were used to treat embryos in the somite, tail-bud, or heart-beating stages for 1 h. Survival rates and rates of normal embryonic development after treatment with trehalose, sucrose, or lactose were all greater than 86.97% and 86.01%, respectively. Levels of toxicity of trehalose, sucrose, and lactose were lower than those of fructose and glucose in C. altivelis embryos. Sucrose and trehalose were combined at different ratios (total concentration 1 M) and used to treat embryos for 1 h. Rates of survival and normal development for the somite, tail-bud, or heart-beating stages did not significantly differ among groups treated with sucrose + trehalose at ratios of 1:1 (S1T1), 1:3 (S1T3), and 3:1 (S3T1). Embryos treated with S1T1 had a little higher survival rate than those of the other treatment ratios(P > 0.05). Survival rates of embryos treated with T1S1, sucrose, and trehalose were 93.86 +/- 4.17%, 91.58 +/- 3.15%, and 92.79 +/- 2.45%, respectively. S1T1 was less toxic than sucrose or trehalose alone under the same conditions. Various cryopreservation-related factors (e.g., the cryoprotectant solution, embryonic stage, equilibrium time, thawing temperature, and time) were screened. The optimal protocol was the treatment of embryos at the tail-bud and heart-beating stages with 1 M S1T1 for 2-10 min and thawing at 26 degrees C for 10 s after freezing. In total, 21 viable embryos were recovered after freezing and thawing, with a survival rate of 0.21-7.55% and maximum survival time of 5 days. Our results demonstrate that it is feasible to cryopreserve C. altivelis embryos using non-permeable cryoprotectants and provide a reference for the use of combinations of cryoprotectants for the species.

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[1]Cryopreservation of embryos of humpback grouper (Cromileptes altivelis) using combinations of non-permeating cryoprotectants. Li, Linlin,Tian, Yongsheng,Li, Zhentong,Li, Ziqi,Duan, Pengfei,Wang, Xinyi,Chen, Shuai,Wang, Linna,Li, Linlin,Tian, Yongsheng,Wang, Linna,Li, Zhentong,Li, Ziqi,Duan, Pengfei,Wang, Xinyi,Chen, Shuai,Wang, Qingbin,Zhai, Jieming. 2022

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