Swine gender-controlling sperm and preparation method and application thereof

A sex and sperm technology, applied in the field of pig sex control sperm and its preparation, can solve the problems of cumbersome cryopreservation method, and achieve the effects of improving vitality and survival time, strong practicability and low dosage

Active Publication Date: 2013-07-31
SHAANXI SHIYANG GRP PUCHENG FARMING DEVCO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this invention can better guarantee the viability, survival rate, plasma membrane integrity rate, acrosome integrity rate and mitochondrial activity of the frozen sperm, the cryopreservation method is relatively cumbersome

Method used

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  • Swine gender-controlling sperm and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Prepare the solution

[0045] 1. Original semen diluent

[0046] Weigh 5.0 g of glucose, 0.3 g of sodium citrate, 0.1 g of disodium edetate, 0.81 mg of genistein (30 μM), 2.42 mg of equol (100 μM), 6.0 g of seminal plasma (fresh boar The supernatant after the semen was centrifuged at 1600r / min) was dissolved in 100 ml of distilled water, and 120,000 international units of penicillin and 0.15 μg of streptomycin were added before use to obtain the original semen dilution (that is, in glucose-sodium citrate - Add genistein, equol and seminal plasma to disodium edetate diluent);

[0047] 2. Freezing diluent A

[0048] Add 20% (volume content) of egg yolk to the original essence dilution, stir well with a magnetic stirrer, centrifuge, take the supernatant and filter, and prepare the frozen dilution A;

[0049] 3. Freezing diluent B

[0050] Add glycerol to the above-mentioned frozen diluent A until the final mass percentage of glycerol is 6%, that is, the frozen dilue...

Embodiment 2

[0061] 1. Prepare the solution

[0062] 1. Original semen diluent

[0063] Weigh glucose 5.0g, sodium citrate 0.3g, edetate disodium 0.1g, genistein 0.54mg (20μM), equol 2.66mg (110μM), seminal plasma 7.0g (fresh boar The supernatant after the semen was centrifuged at 1600r / min) was dissolved in 100 ml of distilled water, and 120,000 international units of penicillin and 0.15 micrograms of streptomycin were added before use to obtain the original semen dilution;

[0064] 2. Frozen diluent

[0065] Adding glycerol to the final mass percentage of glycerin to the above-mentioned original essence dilution is 8%, that is, a frozen dilution is obtained;

[0066] 2. Preparation of porcine sex control spermatozoa

[0067] Collect 2 mL of fresh semen from a healthy boar, and use a fluorescence microscope to detect its viability as 90%; dilute the above-mentioned fresh semen to 400×10 6 Sperm / ml, then add fluorescent dye Hoechst33342 with a concentration of 10mg / mL to a final concen...

Embodiment 3

[0076] 1. Prepare the solution

[0077] 1. Original semen diluent

[0078] Weigh 5.0g of glucose, 0.5g of sodium citrate, 1.35mg of genistein (50μM), 1.94mg of equol (80μM), 7.5g of seminal plasma (the supernatant after centrifuging fresh bull semen at 1600r / min ) was dissolved in 100 ml of distilled water, and 80,000 international units of penicillin and 5 ml of egg yolk were added before use to obtain the original essence dilution; (that is, adding genistein, cow urine phenol and seminal plasma)

[0079] 2. Frozen diluent

[0080] Add glycerol and BHT to the above-mentioned original essence dilution, until the final mass percentage of glycerol is 8%, and the final concentration of BHT is 0.5mM, that is, the frozen dilution is obtained;

[0081] 2. Preparation of porcine sex control spermatozoa

[0082] Collect 2 mL of fresh semen from a healthy boar, and use a fluorescence microscope to detect its viability as 85%; dilute the above-mentioned fresh semen to 300×10 6 Sperm / ...

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Abstract

The present invention discloses a swine gender-controlling sperm and preparation method and application thereof, which comprises diluting the original boar semen with the original semen diluent containing seminal plasma, genistein and cow urine phenol, separating the diluted original boar semen by the flow cytometry and recycling the obtained swine gender-controlling sperm into the freezing diluent containing seminal plasma, genistein and cow urine phenol. The method provided by the present invention has simple operation, which can effectively prevent the separation and the cryopreservation of pig X, Y sperm from damaging, thereby improving the sperm quality and the fertilization ability, and prolonging the survival time of sperm, wherein the separated sperm can be used in low dose insemination, so as to reduce the production cost. The method can be widely applied to commercial promotion with broad market prospects.

Description

technical field [0001] The invention relates to a separation technology of livestock sex-controlling sperm, in particular to a pig sex-controlling sperm and its preparation method and application. Background technique [0002] Animal sex control (sex control) technology is a biological technology that allows adult female animals to produce offspring of the desired sex by artificially intervening in the normal reproductive process of animals. It is of great significance in animal husbandry production. First, by controlling the sex ratio of offspring, the production traits restricted by sex (such as lactation, egg production) and production traits affected by sex (such as growth speed, meat quality, etc.) can be fully utilized, thereby obtaining the greatest economic benefits; secondly, By controlling the sex ratio of the offspring, the intensity of selection can be increased, the breeding process of livestock and poultry can be accelerated, and the breeding and conservation p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C12N5/076A61D19/02
Inventor 祝发明韩春元彭勇翟亚玲贾武利白晓婷同文江
Owner SHAANXI SHIYANG GRP PUCHENG FARMING DEVCO
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