Preparation method and application thereof for dog seminal fluid cryopreservation diluent

A cryopreservation and diluent technology, which is applied in applications, preservation of human or animal bodies, animal husbandry, etc., can solve the problems of low conception rate and litter size, improve survival rate, improve freeze resistance, and inhibit ice crystals the formation of the effect

Inactive Publication Date: 2018-07-20
NANCHANG POLICE DOG BASE MINIST OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the research on the cryopreservation of dog semen has been carried out for many years, the sperm viability after thawing is only about 30%-50%, and the conception rate and litter size after artificial insemination are significantly lower than those of fresh and low-temperature frozen semen

Method used

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  • Preparation method and application thereof for dog seminal fluid cryopreservation diluent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The quality ratio and preparation steps of the four diluent raw materials are as follows:

[0022] Diluent A: Vitamin E 1.2g, glucose 4g, citric acid 1.2g, fructose 2.5g, egg yolk 6g, penicillin 550,000 IU, streptomycin 0.13g, carnitine 0.2g, sodium glutamate 0.7g;

[0023] Diluent B: 3g glucose, 3ml DMSO, 12ml glycerol, 2ml ethylene glycol;

[0024] Diluent C: 0.8g trehalose, 1.5g gelatin, 1.2g EDTA, 0.5g sorbitol, 4g polyvinylpyrrolidone, 1.5g peptone;

[0025] Diluent D: Thimerosal 1.5g, Caspase inhibitor Z-VAD-FMK 4g.

[0026] The preparation methods of the four types of diluents are:

[0027] Diluent A: Put vitamin E, glucose, citric acid, fructose, egg yolk, carnitine, and sodium glutamate in a sterile beaker according to the weight ratio, dissolve in 100ml double distilled water, adjust the pH to 7.2, and stir 8min, then add penicillin and streptomycin according to the weight ratio, stir for 12min, store at 4℃;

[0028] Diluent B: Place glucose, DMSO, glycerol, and ethylene...

Embodiment 2

[0034] The quality ratio and preparation steps of the four diluent raw materials are as follows:

[0035] Diluent A: Vitamin E 1g, glucose 3g, citric acid 1g, fructose 2g, egg yolk 5g, penicillin 500,000 IU, streptomycin 0.1g, carnitine 0.1g, sodium glutamate 0.5g;

[0036] Diluent B: 2g glucose, 2ml DMSO, 10ml glycerol, 1ml ethylene glycol;

[0037] Diluent C: 0.5g trehalose, 1g gelatin, EDTA1, 0.3g sorbitol, 3g polyvinylpyrrolidone, 1g peptone;

[0038] Diluent D: 1.2 g of thimerosal and 3 g of caspase inhibitor Z-VAD-FMK.

[0039] The rest is the same as in Example 1.

Embodiment 3

[0041] The quality ratio and preparation steps of the four diluent raw materials are as follows:

[0042] Diluent A: Vitamin E 1.5g, glucose 5g, citric acid 1.5g, fructose 3g, egg yolk 7g, penicillin 600,000 IU, streptomycin 0.15g, carnitine 0.3g, sodium glutamate 1g;

[0043] Diluent B: 4g glucose, 4ml DMSO, 14ml glycerol, 3ml ethylene glycol;

[0044] Diluent C: 1.2g trehalose, 2g gelatin, 1.5g EDTA, 0.7g sorbitol, 5g polyvinylpyrrolidone, 2g peptone;

[0045] Diluent D: Thimerosal 1.8g, Caspase inhibitor Z-VAD-FMK 5g.

[0046] The rest is the same as Example 1

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Abstract

Provided is a preparation method and application thereof for dog seminal fluid cryopreservation diluent. The diluent comprises diluent A, diluent B, diluent C and diluent D, wherein the diluent A comprises vitamin E, glucose, citric acid, fructose, yolk liquid, carnitine, sodium glutamate, penicillin, and streptomycin; the diluent B comprises glucose, DMSO, glycerinum and glycol; the diluent C comprises mycose, gelatin, EDTA, sorbitol, polyvinyl pyrrolidone and peptone; the diluent D comprises thiomersalate and caspase inhibitor Z-VAD-FMK. According to the prepared diluent, the sperm motilityrate and acrosome intact rate after the dog seminal fluid freezes can be significantly improved.

Description

Technical field [0001] The invention belongs to the technical field of animal reproduction, and specifically relates to a preparation method and application of a dog semen cryopreservation diluent. Background technique [0002] Today, with the rapid development of biotechnology, the semen of some excellent sires is processed into frozen semen through the processing of related technologies, and is vigorously promoted and used through artificial insemination. Among them, the combination of dog frozen semen and artificial insemination technology has a very broad market prospect in the modern kennel industry. Firstly, it can greatly reduce the infection of reproductive system diseases; secondly, it can make full use of excellent male dogs. It is conducive to breed improvement and can save the cost of breeding male dogs. Moreover, since frozen semen can be stored for a long time and transported over long distances, artificial insemination can overcome the breeding difficulties caused ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/021A01N1/0215A01N1/0221A01N1/0226
Inventor 吴衍余盼熊前叶俊华李川武王春亮潘彩霞李涛杨前勇李强
Owner NANCHANG POLICE DOG BASE MINIST OF PUBLIC SECURITY
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