A diluent for cryopreservation of horse sperm

Abstract

The invention provides a low temperature preservation diluent for hose sperm. The low temperature preservation diluent is prepared by fully and uniformly mixing a sugar-salt solution with skimmed milk in proportion of 1:1 in sterile operation, adjusting pH value of the solution to be 6.9 by 1mol / L hydrochloric acid, and finally adjusting the osmotic pressure of the solution to be 300mOsm / L by a 25% glucose solution or double distilled water, wherein the sterile sugar-salt solution is prepared by the steps of sequentially dissolving 0.336-0.374g of sodium bicarbonate, 4.766-4.975g of hydroxyethylpiperazine ethane sulfonic acid (HEPES), 16.213-16.543g of glucose, 45.399-45.423g of lactose, 1.258-1.543g of sodium chloride, 0.348-0.375g of potassium chloride, 0.068-0.086g of monopotassium phosphate, 0.120-0.145g of magnesium sulfate, 0.133-0.154g of calcium chloride, 0.113-0.124g of dibasic sodium phosphate, 2-2.5g of ceftriaxone sodium and 500-600 Mcg of amphotericin B in the 800 ml of double distilled water, then adding the double distilled water to make up to volume of 1,000ml, and filtering the sugar-salt solution with a 0.22-micron filter in an ultra-clean table.

Description

technical field [0001] The invention relates to the preparation of animal sperm cryopreservation solution. Compared with the conventional method, the semen diluent prepared by the method effectively prolongs the survival time of horse sperm in vitro. Background technique [0002] my country is the country with the largest number of horses in the world, with a stock of more than 6 million horses. However, the development of the horse industry is relatively lagging behind. However, at the beginning of the industry's development, how to transform the number advantage of our country's horses into an industrial advantage is a key issue for Chinese horse industry workers. important issues to face. Introducing foreign blood to improve the production performance of Chinese horses on a large scale is an effective method, but the improvement faces two main problems: First, due to the limited storage time of horse semen in vitro, artificial insemination can only be limited to the place ...

AI Technical Summary

Problems solved by technology

Introducing foreign blood to improve the production performance of Chinese horses on a large scale is an effective method, but the improvement faces two main problems: First, due to the limited storage time of horse semen in vitro, artificial insemination can only be limited to the place where the stallion is bred , unable to achieve large-scale, long-distance, cross-regional breed improvement, resulting in the already scarce stallion resources not being effectively utilized

Second, due to the special physiological characteristics of mares, in the process of artificial insemination, semen needs to be transfused 2-3 times at intervals of about 24 hours, while the semen collection frequency of a male horse is generally 72 hours. Due to the limitation, the semen of multiple male horses is used to infuse a single mare in the production, which leads to the confusion of the pedigree of the improved offspring

[0005] The present invention has been working on the diluent for cryopreservation of horse sperm since 2006. It has tried to use defatted mare's milk, Kenney, EZ-Mixin, INRA82 and other diluent formulas for horse sperm in vitro preservation experiments, in vitro preservation time and preservation effect has been continuously improved, but failed to achieve the desired effect

In 2009, we used 100ml double-distilled water, 6.5g glucose, 0.5g glycine, 20% egg yolk, plus penicillin and streptomycin to conduct freezing and cryopreservation experiments on horse sperm. The results at that time showed that "glucose + glycine + "Egg Yolk Diluent" is an alternative to equine sperm cryopreservation, but not suitable for cryopreservation of equine sperm

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