van der Straten K M, Leung L K-P, Rossini R, Johnston S D
School of Animal Studies, The University of Queensland, Gatton, Australia.
Cryo Letters. 2006 Jul-Aug;27(4):203-9.
As a first step towards the development of a method for the cryopreservation of black marlin spermatozoa, this study investigated the effect of dimethylsulfoxide (DMSO) concentration and pellet size on post-thaw spermatozoal motility. Spermatozoa were recovered from the spermatic duct of testes retrieved post-mortem from four adult black marlin caught in the Coral Sea spawning grounds of Australia. Undiluted spermatozoa were stored on ice for 4 to 10 hours during transport to shore, then evaluated for motility after activation in seawater (1:10 v:v). Spermatozoa were prepared for cryopreservation in pellets by extension (1:3 v:v) in a defined fish Ringer's solution to give two final DMSO concentrations of 2.5% or 5.0%. Diluted spermatozoa were frozen directly on a dry ice block in pellet sizes of either 0.25 ml or 0.50 ml. Frozen pellets were thawed in a water bath at 40 degrees C for 60 seconds and assessed for post-thaw motility following activation in seawater. Spermatozoa recovered within 50 minutes of death and chilled on ice for 4 to 10 hours showed a mean (+/- SEM) motility immediately following activation of 91.6 +/- 7.9%. 50% of the spermatozoa remained motile for approximately 4 to 5 minutes. Following cryopreservation, mean motility declined significantly across all cryoprotectant and pellet size combinations (P < 0.001) but spermatozoa frozen in 2.5% DMSO showed higher motility than those frozen in 5.0% DMSO (P = 0.014). Pellet size had no effect on post-thaw motility (P = 0.179).
作为开发黑枪鱼精子冷冻保存方法的第一步，本研究调查了二甲基亚砜（DMSO）浓度和颗粒大小对解冻后精子活力的影响。从在澳大利亚珊瑚海产卵场捕获的4条成年黑枪鱼死后取出的睾丸输精管中回收精子。未稀释的精子在运往岸边的过程中在冰上保存4至10小时，然后在海水中激活（1:10 v:v）后评估活力。通过在特定的鱼类林格氏液中按1:3 v:v的比例稀释制备用于冷冻保存的精子颗粒，使最终的DMSO浓度为2.5%或5.0%。稀释后的精子直接在干冰块上冷冻成0.25 ml或0.50 ml的颗粒大小。冷冻颗粒在40摄氏度的水浴中解冻60秒，并在海水中激活后评估解冻后的活力。在死亡后50分钟内回收并在冰上冷藏4至10小时的精子在激活后的平均（±SEM）活力为91.6±7.9%。50%的精子保持活力约4至5分钟。冷冻保存后，所有冷冻保护剂和颗粒大小组合的平均活力均显著下降（P<0.001），但在2.5%DMSO中冷冻的精子比在5.0%DMSO中冷冻的精子活力更高（P = 0.014）。颗粒大小对解冻后活力没有影响（P = 0.179）。
