Cryopreservation of embryos of humpback grouper (Cromileptes altivelis) using combinations of non-permeating cryoprotectants
文献类型: 外文期刊
作者: Li, Linlin 1 ; Tian, Yongsheng 1 ; Li, Zhentong 1 ; Li, Ziqi 1 ; Duan, Pengfei 1 ; Wang, Xinyi 1 ; Chen, Shuai 1 ; Wang, Linna 1 ; Wang, Qingbin 5 ; Zhai, Jieming 5 ;
作者机构: 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Minist Agr & Rural Affairs, Key Lab Sustainable Dev Marine Fisheries, Qingdao 266071, Peoples R China
2.Chinese Acad Agr Sci, Grad Sch, Beijing 100081, Peoples R China
3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266000, Peoples R China
4.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai, Peoples R China
5.Laizhou Mingbo Aquat Co Ltd, Yantai 2614000, Peoples R China
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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