Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8464068 | Cryobiology | 2018 | 24 Pages |
Abstract
We investigated various factors, including cryoprotective agents (CPAs), diluents, and freezing rates, to develop an optimal cryopreservation protocol for Epinephelus akaara sperm. In experiments using 10% dimethyl sulfoxide (DMSO), glycerol, and methanol with various diluents, 10% DMSO and 300â¯mM glucose yielded the highest post-thaw sperm motility. The combination of 10% glycerol and 300â¯mM sucrose yielded significantly higher post-thaw sperm motility than did combinations using other diluents. Glycerol and DMSO at a concentration of 10% as CPAs with 300â¯mM glucose as the diluent resulted in the highest MSR and sperm activity index (SAI). An investigation to determine the effects of glycerol and DMSO concentrations on post-thaw sperm survival rate revealed no significant differences among 5, 10, 15, and 20% concentrations of either CPA. In assessing the effects of CPA concentration on the fertilization rate, the 10% concentration yielded the highest fertilization rate (81.4â¯Â±â¯4.3%) in DMSO, whereas 15% was the optimal concentration for glycerol (fertilization rateâ¯=â¯66.7â¯Â±â¯6.1%). The hatching rate was also highest in 10% DMSO (40.1â¯Â±â¯0.4%) and in 15% glycerol (27.8â¯Â±â¯2.3%). In conclusion, the optimal rates of post-thaw sperm motility, fertilization, and hatching were achieved when E. akaara sperm were cryopreserved in a diluent of 300â¯mM glucose with 10% DMSO as the CPA at a freezing rate of â5â¯Â°C/min. We therefore recommend this protocol for the cryopreservation of E. akaara sperm.
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Authors
Jae Yeon Ahn, Jung Yeol Park, Han Kyu Lim,