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Cattle freezing seminal fluid dilution and method for producing the same

A technology of freezing semen and diluent, which is applied in the biological field, can solve the problems of low viability and achieve the effect of improving semen quality and survival rate

Inactive Publication Date: 2008-07-16
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the defect that the viability of bovine sperm is relatively low after freezing in my country at present, the purpose of the present invention is to provide a diluent of bovine frozen semen, which can achieve the effect of 50% to 68% viability of frozen bovine semen after thawing, greatly improving Improve the production efficiency of bovine frozen semen in my country

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The extraction process of embodiment 1 low-density lipoprotein:

[0022] After fresh egg yolks are collected, first dilute the egg yolks by 2 to 3 times with isotonic saline solution (0.17M NaCl, w / v), and keep stirring for 1 hour, then centrifuge at 10,000×g for 45 minutes at 4°C, and the supernatant The liquid is separated from the sediment. In order to avoid the interference of small particles contained in the egg yolk, the separated supernatant was centrifuged again under the same conditions. The supernatant collected after the second centrifugation was mixed with 40% ammonium sulfate solution, and after being fully stirred at 4°C for 1 hour, the mixture was centrifuged for 45 minutes (10,000×g, 4°C) to separate the layers to precipitate the egg yolk protein. After centrifugation, the precipitate was discarded, and the supernatant (floating matter) was dialyzed with distilled water for 12 hours to remove ammonium sulfate in the solution. After the ammonium sulfat...

Embodiment 2

[0023] Embodiment 2 The preparation method of bovine frozen semen dilution

[0024] Accurately weigh 1.0g of fructose, 1.48g of sodium citrate, and 2.42g of TRIS with an electronic analytical balance, dissolve them in 50ml of distilled water, stir them evenly with a magnetic stirrer, and prepare 100ml of base solution. Add 100,000 IU of penicillin G sodium and 8 g of low-density lipoprotein to the base solution to prepare liquid I; add 6.4 ml of glycerin to liquid I to prepare liquid II. Then use a precision pH meter to adjust the pH to 6.37, then filter with a 0.22 μm filter membrane, and put it in a refrigerator at 2-5°C for later use. I and II liquids need to be prepared immediately after use.

Embodiment 3

[0025] Embodiment 3 The preparation method of bovine frozen semen dilution

[0026] Accurately weigh 0.5g of fructose, 2g of sodium citrate, and 1.5g of TRIS with an electronic analytical balance, dissolve them in 50ml of distilled water, stir them evenly with a magnetic stirrer, and prepare 100ml of base solution. Add 50,000 IU of penicillin G sodium and 5 g of low-density lipoprotein to the base solution to prepare liquid I; add 8 ml of glycerin to liquid I to prepare liquid II. Then use a precision pH meter to adjust the pH to 6.4, then filter with a 0.22 μm filter membrane, and put it in a refrigerator at 2-5°C for later use. I and II liquids need to be prepared immediately after use.

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Abstract

The invention discloses a frozen bovine semen diluent, and the consumption amounts of all the raw materials in 100ml of the diluent are: 0.8 to 1.2g of fructose, 1.4 to 1.6g of sodium citrate, 2.3 to 2.6g of TRIS, 7.5 to 9.5g of low-density lipoprotein, 5 to 8ml of glycerol, 0.085 to 0.12 million IU of penicillin and the rest is distilled water. The preparation method is that: the fructose, the sodium citrate and the TRIS are dissolved in the distilled water to be prepared into a base liquid; penicillin G sodium and the low-density lipoprotein are added in the base liquid to prepare a I liquid; the glycerol is added in the I liquid to prepare a II liquid; then the pH value is adjusted to 6 to 7.5, then a filtration and a sterilization are carried out, and the liquid is cooled until reaching the room temperature and then arranged in a refrigerator of 2 - 5 DEG C for standby. The frozen bovine semen diluent of the invention has good and reliable effect, which can provide high-quality and excellent straw frozen semen for bovine artificial insemination and have very broad market application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to ultra-low temperature cryopreservation of animal semen, in particular to a bovine frozen semen dilution and a preparation method thereof. Background technique [0002] The combination of bovine semen cryopreservation and artificial insemination technology plays an extremely important role in the production of modern cattle breeding. The application of frozen semen for artificial insemination can not be limited by time, region, cattle breed and individual physique, and can greatly reduce the risk of bulls. Feed quantity, reduce production cost, improve the utilization rate of good breeding bulls, and bring huge economic benefits to the cattle industry. At the same time, the development of bovine semen freezing technology and the use of bovine frozen semen are of great significance in the introduction of bovine breeds, hybrid improvement, improvement of reproductive performance, disease ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06A01N1/02
Inventor 李青旺胡建宏赵宏伟江中良
Owner NORTHWEST A & F UNIV
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