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用含1%或2%大豆卵磷脂替代蛋黄的Tris稀释液对犬精液进行冷冻保存。

Cryopreservation of Dog Semen in a Tris Extender with 1% or 2% Soya Bean Lecithin as a Replacement of Egg Yolk.

作者信息

Axnér E, Lagerson E

机构信息

Division of Reproduction, Department of Clinical Sciences, Faculty of Veterinary Medicine and Animal Health, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.

出版信息

Reprod Domest Anim. 2016 Apr;51(2):262-8. doi: 10.1111/rda.12675. Epub 2016 Mar 6.

Abstract

Egg yolk is usually included in extenders used for preservation of dog semen. Lecithin is an interesting animal-protein free alternative to egg yolk for semen preservation. The aim of our study was to evaluate soya bean lecithin for cryopreservation of dog semen. Five ejaculate replicates were divided in three equal parts, centrifuged and each pellet diluted with one of the three Tris-based extenders containing 20% egg yolk, 1% soya bean lecithin or 2% soya bean lecithin. Extended semen was loaded in 0.5-ml straws, cooled and diluted a second time and frozen in liquid nitrogen vapours. Sperm motility parameters (CASA), acrosome integrity (FITC-PNA/PI) and sperm membrane integrity (C-FDA) were evaluated 5 min post-thaw and after 2 and 4 h of incubation. Total motility was significantly better in the egg yolk extender than in any of the lecithin-based extender and was better in the 1% lecithin extender than in the 2% lecithin extender. Sperm membrane integrity was significantly better in the egg yolk extender than in any of the lecithin-based extenders but did not differ significantly between the 1% and 2% lecithin extenders. Acrosome integrity was significantly better in the egg yolk extender than in the 2% lecithin extender but did not differ between the egg yolk extender and the 1% lecithin extender or between the two lecithin extenders. In conclusion, egg yolk was superior to lecithin in our study. The extender with 1% lecithin preserved sperm motility better than the extender with 2% lecithin.

摘要

蛋黄通常包含在用于保存犬精液的稀释液中。卵磷脂是一种有趣的、不含动物蛋白的蛋黄替代物,可用于精液保存。我们研究的目的是评估大豆卵磷脂用于犬精液冷冻保存的效果。将五次射精的样本各分成三等份,进行离心,每个沉淀用三种基于Tris的稀释液之一进行稀释,这三种稀释液分别含有20%蛋黄、1%大豆卵磷脂或2%大豆卵磷脂。将稀释后的精液装入0.5毫升细管中,冷却并再次稀释,然后在液氮蒸汽中冷冻。解冻后5分钟以及孵育2小时和4小时后,评估精子活力参数(计算机辅助精子分析)、顶体完整性(异硫氰酸荧光素 - 花生凝集素/碘化丙啶)和精子膜完整性(羧基荧光素二乙酸酯)。蛋黄稀释液中的总活力显著高于任何一种基于卵磷脂的稀释液,1%卵磷脂稀释液中的总活力优于2%卵磷脂稀释液。蛋黄稀释液中的精子膜完整性显著优于任何一种基于卵磷脂的稀释液,但1%和2%卵磷脂稀释液之间没有显著差异。蛋黄稀释液中的顶体完整性显著优于2%卵磷脂稀释液,但在蛋黄稀释液与1%卵磷脂稀释液之间或两种卵磷脂稀释液之间没有差异。总之,在我们的研究中,蛋黄优于卵磷脂。含1%卵磷脂的稀释液比含2%卵磷脂的稀释液能更好地保存精子活力。

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