Diluent of frozen semen of milk goat, and preparation method and diluting method thereof as well as preparation method of frozen semen of milk goat in thin pipe

A technology of frozen diluent and frozen semen, which is applied in the biological field and can solve the problems of large-scale production that is difficult to increase the conception rate, changes, and affects the sperm viability and fertilization ability of the plasma membrane integrity.

Inactive Publication Date: 2015-04-01
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The application of frozen semen technology and artificial insemination technology in dairy cows and beef cattle in my country has been relatively well developed, which has fully improved the economic benefits of breeding. However, although goat semen cryopreservation and artificial insemination technology have been used for many years, the frozen semen is generally frozen after thawing. , the sperm motility rate (0.3-0.5) is difficult to increase the conception rate (35%-50%) to meet the needs of large-scale production
The reason is that the structure of the plasma membrane of the sperm is changed by the shock of the sperm, which seriously affects the integrity of the plasma membrane and the viability and fertilization ability of the recovered sperm.

Method used

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  • Diluent of frozen semen of milk goat, and preparation method and diluting method thereof as well as preparation method of frozen semen of milk goat in thin pipe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0166] Step 1: Prepare base dilution:

[0167] (1) Prepare the components according to the following proportions:

[0168] Tris-aminomethane 300 mM;

[0169] Citric acid 100 mM;

[0170] Glucose 35mM;

[0171] Trehalose 260 mM;

[0172] Vitamin C 4 g / L;

[0173] Reduced glutathione 4 mM;

[0174] Bovine serum albumin 8 g / L;

[0175] Adenosine triphosphate 0.7 mM;

[0176] (2) Mix the above components with double-distilled water in proportion, filter through a 0.2 um filter membrane, and add compound antibiotics, that is, add 0.05 g / L tylosin, 0.27 g / L gentamicin, 0.16 g / L of lincomycin and 0.3 g / L of spectinomycin;

[0177] (3) According to the ratio of 15% by volume, add egg yolk additive solution to the mixed solution obtained in step 2, and mix well to obtain the basic dilution solution;

[0178] The egg yolk supplement is prepared by the following steps:

[0179] Fresh eggs free of pathogenic bacteria were soaked in chlorhexidine to extract yolk liquid, and sodiu...

Embodiment 2

[0189] Step 1: Prepare base dilution:

[0190] (1) Prepare the components according to the following proportions:

[0191] Tris-aminomethane 350 mM;

[0192] Citric acid 115 mM;

[0193] Glucose 40mM;

[0194] Trehalose 280 mM;

[0195] Vitamin C 5 g / L;

[0196] Reduced glutathione 5 mM;

[0197] Bovine serum albumin 10 g / L;

[0198] Adenosine triphosphate 1.1 mM;

[0199] (2) Mix the above components with double-distilled water in proportion, filter through a 0.2 um filter membrane, and add compound antibiotics, that is, add 0.30 g / L tylosin, 0.27 g / L gentamicin, 0.17 g / L of lincomycin and 0.30 g / L of spectinomycin;

[0200] (3) According to the ratio of 20% by volume, add egg yolk additive solution to the mixed solution obtained in step 2, and mix well to obtain the basic dilution solution;

[0201] The egg yolk supplement is prepared by the following steps:

[0202] Fresh eggs free of pathogenic bacteria were soaked in chlorhexidine to extract yolk liquid, and sodiu...

Embodiment 3

[0212] Step 1: Prepare base dilution:

[0213] (1) Prepare the components according to the following proportions:

[0214] Tris-aminomethane 400 mM;

[0215] Citric acid 130 mM;

[0216] Glucose 45mM;

[0217] Trehalose 300 mM;

[0218] Vitamin C 6 g / L;

[0219] Reduced glutathione 6 mM;

[0220] Bovine serum albumin 12 g / L;

[0221] Adenosine triphosphate 1.5 mM;

[0222] (2) Mix the above components with double-distilled water in proportion, filter through a 0.2 um filter membrane, and add compound antibiotics, that is, add 0.055 g / L tylosin, 0.28 g / L gentamicin, 0.18 g / L of lincomycin and 0.35 g / L of spectinomycin;

[0223] (3) According to the ratio of 25% by volume, add egg yolk additive solution to the mixed solution obtained in step 2, and mix well to obtain the basic dilution solution;

[0224] The egg yolk supplement is prepared by the following steps:

[0225] Fresh eggs free of pathogenic bacteria were soaked in chlorhexidine to extract yolk liquid, and so...

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Abstract

The invention relates to dairy goat frozen semen diluent, its preparation method and dilution method, and a preparation method of dairy goat frozen semen thin tube frozen semen. In the existing frozen semen preservation method, the plasma membrane structure of the sperm is changed due to the impact of low temperature, which seriously affects the integrity of the plasma membrane and its viability and fertilization ability after recovery. The present invention performs two-step dilution, balance and two-step cooling thin tube freezing treatment on fresh semen, wherein, the basic diluent is composed of Tris, citric acid, glucose, trehalose, Vc, GSH, BSA, ATP, compound antibiotics, egg yolk and water The frozen diluent is made up of basic diluent plus glycerin; after the diluent is used to dilute and balance the fresh semen, it is divided into thin tubes and stored in liquid nitrogen. The invention has good effect on cryopreservation and cryopreservation of dairy goat semen, effectively improves the survival rate after thawing, the integrity rate of acrosome and the integrity of plasma membrane, reduces the deformity rate, and improves the production efficiency and quality of frozen semen of dairy goats.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a dairy goat frozen semen diluent, a preparation method and a dilution method thereof, and a preparation method of dairy goat frozen semen thin tube frozen semen. Background technique [0002] Dairy goats, as an important dairy animal resource, have received much attention in recent years with the improvement of the living standard of our country and the adjustment of diet structure. Dairy goat breeding is mainly concentrated in Shaanxi, Shandong, Henan, Hebei, Yunnan and other provinces. However, the demand for goat milk has spread to more than 10 provinces, cities, autonomous regions and municipalities in the eastern coastal areas and southern Hong Kong, Macao and Taiwan, reflecting a certain Therefore, how to speed up the improvement of dairy goat breeds, improve the breeding speed of dairy goat fine breeds and establish a large-scale breeding management system has become th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
Inventor 罗军王维史怀平席利萌孙爽高庆华张伟杨地坤朱江江
Owner NORTHWEST A & F UNIV
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