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脊尾白虾精子体外保存的研究
浦蕴惠1,2, 许星鸿2, 高 焕2, 刘兆普1, 阎斌伦2
1.南京农业大学 资源与环境科学学院 江苏省海洋生物学重点实验室;2.淮海工学院江苏省海洋生物技术重点建设实验室
摘要:
采用精子存活率和顶体酶活力两种评价精子质量方法, 研究了不同稀释剂与抗冻保护剂对脊尾白虾(Exopalaemon carinicauda)精子体外保存效果的影响。结果表明: 稀释剂Ⅰ-壬氏液为脊尾白虾精子体外保存的最佳稀释剂, 4℃下保存4 d 后, 稀释剂Ⅰ组精子存活率为71.50 %, 顶体酶活力为1.803μ IU/106, 两种指标值均高于其他实验组, 各组精子存活率与顶体酶活力之间相关性显著γ=0.846(P<0.05); 在液氮-196℃下保存2 d 后, 稀释剂Ⅰ组精子存活率达83.50 %, 顶体酶活力为2.507μIU/106, 两种指标值均高于其他各组以及4℃下稀释剂Ⅰ组同期保存效果, 各组精子存活率与顶体酶活力之间相关性极显著, γ=0.862 (P<0.01)。15%DMSO 为脊尾白虾精子超低温保存的最佳抗冻保护剂,此条件下超低温保存2 d 后, 精子存活率为81.32%, 顶体酶活力为1.385μIU/106, 各组精子存活率与顶体酶活力之间相关性极显著, 相关系数γ=0.864(P<0.01)。
关键词:  脊尾白虾(Exopalaemon carinicauda)  精子  体外保存  稀释剂  抗冻剂
DOI:
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基金项目:国家“十二五”科技支撑计划资助项目(2011BAD13B03)
Study on in vitro preservation of spermatozoa of Exopalaemon carinicauda
Abstract:
In this paper, the effect of different diluting solutions and cryoprotectants on in vitro cryopreservation of spermatozoa was studied by evaluation of spermatozoa quality using survival rate and acrosin activity assays. The results showed that Ringer’s extender was the best diluting solution of spermatozoa in vitro preservation. After a preservation for 4 d at 4℃ with Ringer’s extender, the survival rate was 71.50%, and the acrosin activity reduced to 1.803μIU/106. Both of these two values were higher than those of other experimental groups. The survival rate was significantly correlated with acrosin activity in all the experimental groups, R=0.846 (P<0.05). After a preservation for 2 d in liquid nitrogen with Ringer’s extender, the survival rate was 83.50% and the acrosin activity reduced to 2.507μIU/106. Similarly, both of them were higher than other groups, as well as the values obtained at 4℃. There existed a highly significant correlation between survival rate and acrosin activity, R=0.862 (P<0.01). 15% DMSO was the best cryoprotectant for cryopreservation. After a 2 d preservation, the survival rate was 81.32%, and the acrosin activity reduced to 1.385μIU/106. There existed a highly significant correlation between survival rate and acrosin activity, R=0.864 (P<0.01).
Key words:  Exopalaemon carinicauda  spermatozoa  in vitro preservation  diluting solution  cryoprotectants
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