不同缓冲液添加大豆蛋白冷冻奶牛精子试验效果研究

Study on the Effect of Different Buffer Addition of Soybean Protein on Freezing Bovine Spermatozoa

本文为解决奶牛冻精稀释液添加卵黄易污染、易将异源动物体内微生物传染给输精母畜等弊端,进行了两个方面的试验研究:(1)选择含蛋白量高的大豆粉溶液添加Tris缓冲液制成蛋白Tris稀释液,添加磷酸盐缓冲液制成蛋白PBS稀释液,及传统使用的卵黄Tris稀释液,分别冷冻奶牛鲜精,观察解冻后活力;(2)观察蛋白Tris稀释液、蛋白PBS稀释液与卵黄Tris稀释液超低温冷冻奶牛精子解冻后在4℃环境中存活时间。结果表明,蛋白Tris稀释液、蛋白PBS稀释液与卵黄Tris稀释液超低温冷冻的奶牛,细管冻精质量都能符合国家标准,但冷冻效果最佳的是蛋白Tris稀释液。在冷藏4℃下,蛋白Tris稀释液、卵黄Tris稀释液第5天精子活力仍能达到国家标准,蛋白Tris稀释液显微镜下的观察视野要比卵黄Tris稀释液清晰,不易滋长细菌。因此,添加大豆粉溶液制成的蛋白Tris稀释液冷冻奶牛精子的效果最佳,试验证明,植物源稀释液可以替代动物源稀释液。

In this paper, in order to solve the problems that egg diluent of bovine spermatozoa frozen semen is liable to pollut by heterologous microorganisms, and infectious to insemination dams, this study includes 2 experiments （1） Tris protein dilution is made from high protein content of soybean powder solution with Tris buffer added, PBS protein dilution is made from high protein content of soybean powder solution with phosphate buffer solution, and the traditional liquid is yolk Tris diluent, they were prepared for bovine semen frozen.（2） To observe Tris protein diluent, protein PBS diluent and yolk Tris diluent cryopreservation of bovine sperm thawing in 4~C environment preservation time. The experimental results show that Tris protein diluent, PBS protein diluent and yolk Tris diluent have no significant difference in liquid nitrogen of frozen bovine spermatozoa. Tris protein diluent and yolk Tris diluent can reach national standard of sperm motility for five days in 4~C environment preservation time. Tris protein diluent can easily distinct sperm motility and exam under microscope compared to yolk Tris diluent, it is not easily to develop bacteria. So, the effect in freezing experiment and survival time experiment of Tris protein diluent is the best, plant source dilution can substitute animal source dilution for bovine semen frozen.