Freezing mixed sperm for controlling X/Y gender of milk cattle and preparation method thereof

A technology for freezing semen and semen, which can be used in sexual diseases, drug combinations, and preservation of human or animal bodies. cost effect

Inactive Publication Date: 2009-04-22
李喜和
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the X / Y sperm separation technology based on the difference in DNA content has been popularized and applied in the field of livestock breeding, the cost of each sperm separator is very expensive, the technical operation is complicated, and the speed of X / Y sperm separation is still very limited (5000- 6000 sperm per second), so that the production cost of dairy cow sex control frozen semen is high, and it cannot be popularized and applied in large quantities

Method used

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  • Freezing mixed sperm for controlling X/Y gender of milk cattle and preparation method thereof

Examples

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Effect test

Embodiment 1

[0050] The following examples can help those skilled in the art to understand the present invention more comprehensively. But it does not limit the present invention in any way, and the number of parts in the embodiments of the present invention can be selected as grams or kilograms according to actual needs. Example 1: Production of frozen semen and artificial insemination of 1 million / 0.25ml dairy cow X sex control

[0051] 1) X / Y sperm separation:

[0052] ①Collect 3.6ml of fresh dairy bull semen, and use an 80um sperm filter to remove impurities in the original semen. The concentration of the original semen is 940 million / ml, and the vitality of the original semen is 85%;

[0053] ② Dilute the original semen to 100 million / ml with sperm staining solution TALP, take 4ml of the diluted semen and add 10ul of 15mg / ml sperm DNA fluorescent dye Hochist33342 (purchased from Sigma, USA), and incubate in a warm water bath at 24°C 20 minutes, while adding 2.5% food red dyed dead s...

Embodiment 2

[0073] Example 2: Frozen semen production and artificial insemination of 500,000 / 0.25ml dairy cow X sex control

[0074] 1) X / Y sperm separation:

[0075] ①Collect 3.2ml of fresh dairy bull semen, and use an 80um sperm filter to remove impurities in the original semen. The concentration of the original semen is 800 million / ml, and the vitality of the original semen is 88%;

[0076] ② Dilute the original semen to 100 million / ml with sperm staining solution TALP, take 4ml of the diluted semen and add 10ul of 15mg / ml sperm DNA fluorescent dye Hochist33342, and incubate in a warm water bath at 24°C for 20 minutes (add an appropriate amount of food red and remove stained dead sperm);

[0077] ③According to the operating procedure of the SX-MoFlo sperm separator, implant the dyed sperm sample into the sample table, jet the sperm at a speed of 20,000 sperm per second, and the X sperm separation speed is stable at 5500-6000 per second;

[0078] ④ At the speed of recovering 12 millio...

Embodiment 3

[0096] Example 3: 1 million / 0.25ml beef cattle Y sex control frozen semen production and artificial insemination

[0097] 1) X / Y sperm separation:

[0098] ①Collect 2.8ml of fresh beef bull semen, remove impurities in the original semen with an 80um sperm filter, the concentration of the original semen is 1.48 billion / ml, and the vitality of the original semen is 90%;

[0099]② Dilute the original semen to 100 million / ml with sperm staining solution TALP, take 4ml of the diluted semen and add 10ul of 15mg / ml sperm DNA fluorescent dye Hochist33342, and incubate in a warm water bath at 24°C for 20 minutes (add an appropriate amount of food red and remove stained dead sperm);

[0100] ③In accordance with the operating procedure of the SX-MoFlo sperm separator, implant the dyed sperm sample into the sample table, and jet the sperm at a speed of 20,000 sperm per second, and the Y sperm separation speed is stable at 6000-7000 per second;

[0101] ④ At the speed of recovering 12 mi...

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Abstract

The invention discloses a new technology for producing frozen mixed semen for controlling the gender of milk cow. Goat semen is used as foundation (2,000,000 to 3,000,000 pieces / 0.25 ml) and is added with X / Y separation semen (500,000 to 1,000,000 pieces / 0.25 ml) to produce the frozen semen for controlling the gender X / Y of the milk cow. The main content of the technology comprises the separation of milk cow X / Y sperms, the preparation of the basic goat semen (which can be substituted by the semen of pig, horse or deer with the equal number) and the production of the frozen semen for controlling the gender of the milk cow through the procedures of mixing and freezing two kinds of the semen according to requirements. The new technology can improve the production efficiency of the frozen semen for controlling the gender of the milk cow by 2 to 4 times and reduce the production cost by 50 to 70 percent simultaneously. The new technology improves the activity and the acrosome intact rate of sperms after the sperms are unfrozen, prolongs the time for having the insemination capacity after the frozen sperms are unfrozen so that the average pregnancy rate in a sexual period reaches over 50 percent; and the accuracy rate of gender control reaches over 90 percent. The new technology is also suitable for producing the frozen mixed semen for controlling the gender of beef cattle.

Description

technical field [0001] The invention belongs to the technical field of livestock artificial insemination, and relates to a new technology for the production of milk cow gender-controlled frozen semen, more specifically, milk cow X / Y gender-controlled frozen mixed semen and a preparation method thereof. Background technique [0002] Mammalian sex control has always been one of the important research topics in life sciences. With the development of molecular biology, cytogenetics, developmental biology, immunohistochemistry and other disciplines, people have a better understanding of the mechanism of sex determination . Gender control technology refers to the biotechnology that makes adult female animals produce offspring of a specific sex according to people's wishes by artificially intervening and manipulating the normal reproduction of animals. Controlling the sex of mammals to produce more offspring of the desired sex has very important theoretical and practical significa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/52A01N1/02A61P15/00
CPCA61K35/52A61P15/00
Inventor 李喜和周文忠钱松晋王建国张元炜
Owner 李喜和
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