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斑点叉尾鮰精子的冷冻保存:冷冻保护剂、细管规格及稀释液配方的影响

Cryopreservation of channel catfish spermatozoa: effect of cryoprotectant, straw size, and formulation of extender.

作者信息

Christensen J M, Tiersch T R

机构信息

School of Forestry, Wildlife, and Fisheries, Louisiana Agricultural Experiment Station, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA.

出版信息

Theriogenology. 1997 Feb;47(3):639-45. doi: 10.1016/s0093-691x(97)00022-8.

Abstract

Various aspects of the cryopreservation of spermatozoa of channel catfish (Ictalurus punctatus ) were studied in relation to spermatozoa motility. The objectives were to evaluate 1) the efficacy of 5, 10 and 15% of methanol or n,n-dimethyl acetamide (DMA) as cryoprotectants; 2) the acute toxicity of 5, 10 and 15% methanol or DMA; 3) the use of 0.5-ml vs. 0.25-ml straws; 4) the efficacy of 5, 10 and 15% of methanol in Hanks' balanced salt solution (HBSS) or HBSS without glucose, and 5) the use of HBSS with or without 5% methanol. We found that use of 5% methanol as a cryoprotectant resulted in significantly higher post-thaw motility (P = 0.0001) than did 5, 10 or 15% DMA. The use of 5% of either cryoprotectant resulted in significantly higher post-thaw motility (P = 0.0001) than did 10 or 15% of the cryoprotectants. Samples containing 10 or 15% DMA had significantly lower motility (P = 0.0001) after 30 min exposure than did samples containing 5, 10 or 15% methanol. The use of 0.25-ml straws resulted in significantly higher post-thaw motility (P = 0.0001) than that of 0.5-ml straws. No difference was found in post-thaw motility between HBSS with and without glucose as the extenders. Cryopreservation in HBSS without addition of cryoprotectant resulted in post-thaw motility values of about 1%.

摘要

针对斑点叉尾鮰精子的冷冻保存的各个方面,研究了其与精子活力的关系。目的是评估:1)5%、10%和15%的甲醇或N,N -二甲基乙酰胺(DMA)作为冷冻保护剂的效果;2)5%、10%和15%甲醇或DMA的急性毒性;3)0.5毫升与0.25毫升细管的使用;4)5%、10%和15%甲醇在汉克斯平衡盐溶液(HBSS)或不含葡萄糖的HBSS中的效果;5)含或不含5%甲醇的HBSS的使用。我们发现,与5%、10%或15%的DMA相比,使用5%甲醇作为冷冻保护剂解冻后的活力显著更高(P = 0.0001)。使用5%的任何一种冷冻保护剂解冻后的活力都显著高于10%或15%的冷冻保护剂(P = 0.0001)。暴露30分钟后,含10%或15% DMA的样本活力显著低于含5%、10%或15%甲醇的样本(P = 0.0001)。使用0.25毫升细管解冻后的活力显著高于0.5毫升细管(P = 0.0001)。作为稀释剂,含葡萄糖和不含葡萄糖的HBSS解冻后的活力没有差异。在不添加冷冻保护剂的HBSS中进行冷冻保存,解冻后的活力值约为1%。

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