Casein anti-freezing agent for cryopreservation of donkey semen and preparation method thereof

A technology for cryopreservation and casein, which is applied in the field of casein antifreeze and its preparation, can solve the problems of long reproductive cycle of female animals, low artificial insemination fertilization rate, complicated production and operation of frozen semen of donkeys, etc.

Active Publication Date: 2013-04-17
SHANDONG TIANLONG ANIMAL HUSBANDRY SCI & TECHNOLEGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention proposes a casein antifreeze agent for donkey semen cryopreservation, which solves the problems in the prior art that the reproducti...

Method used

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  • Casein anti-freezing agent for cryopreservation of donkey semen and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0067] Each 100mL of antifreeze contains 1.0g of casein, 2.0g of glucose, 1.0g of lactose, 3.0g of HEPESBUFFER, 2.0mL of glycerin, 2mL of egg yolk, 450,000 IU of penicillin, 4.5g of gentamicin, and 20mg of amphotericin B. The balance is HANK solution, weigh each component and set aside;

[0068] 2) Prepare the above-weighed casein, glucose, lactose, and HEPES BUFFER with HANK solution to form a base solution;

[0069] 3) Add the penicillin, gentamicin, and amphotericin B weighed in step 1) to the base liquid prepared above to make liquid I;

[0070]4) Adjust the pH of the liquid I to 7.1, then filter and sterilize it, and put it in a refrigerator at 3°C ​​for later use;

[0071] 5) Take the liquid I before use, add the egg yolk weighed in step 1) to it, and prepare liquid II to obtain the casein antifreeze agent for cryopreservation of donkey semen.

Embodiment 2

[0073] Each 100mL of antifreeze contains 2.0g of casein, 2.5g of glucose, 1.5g of lactose, 5.0g of HEPESBUFFER, 2.5mL of glycerin, 3mL of egg yolk, 500,000 IU of penicillin, 5g of gentamicin, 25mg of amphotericin B, and the rest The amount is the HANK solution, weigh each component and set aside;

[0074] 2) Prepare the above-weighed casein, glucose, lactose, and HEPES BUFFER with HANK solution to form a base solution;

[0075] 3) Add the penicillin, gentamicin, and amphotericin B weighed in step 1) to the base liquid prepared above to make liquid I;

[0076] 4) Adjust the pH of the liquid I to 7.5, then filter and sterilize it, and put it in a refrigerator at 4°C for later use;

[0077] 5) Take the liquid I before use, add the egg yolk weighed in step 1) to it, and prepare liquid II to obtain the casein antifreeze agent for cryopreservation of donkey semen.

Embodiment 3

[0079] Each 100mL of antifreeze contains 3.0g of casein, 3.0g of glucose, 2.0g of lactose, 6.0g of HEPESBUFFER, 3.0mL of glycerin, 4mL of egg yolk, 550,000 IU of penicillin, 5.5g of gentamicin, and 30mg of amphotericin B. The balance is HANK solution, weigh each component and set aside;

[0080] 2) Prepare the above-weighed casein, glucose, lactose, and HEPES BUFFER with HANK solution to form a base solution;

[0081] 3) Add the penicillin, gentamicin, and amphotericin B weighed in step 1) to the base liquid prepared above to make liquid I;

[0082] 4) Adjust the pH of the liquid I to 7.8, then filter and sterilize it, and put it in a refrigerator at 5°C for later use;

[0083] 5) Take the liquid I before use, add the egg yolk weighed in step 1) to it, and prepare liquid II to obtain the casein antifreeze agent for cryopreservation of donkey semen.

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Abstract

The invention provides a casein anti-freezing agent for cryopreservation of donkey semen and a preparation method thereof. Each 100 milliliters of anti-freezing agent contains the following components: 1.0 to 3.0 grams of casein, 2.0 to 3.0 grams of glucose, 1.0 to 2.0 grams of lactose, 3.0 to 6.0 grams of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) BUFFER, 2.0 to 3.0 milliliters of glycerol, 2 to 4 milliliters of yolk, 450 to 550 thousand international units (IU) of penicillin, 4.5 to 5.5 grams of gentamicin, 20 to 30 milligrams of amphotericin B, and the balance of HANK solution. The casein anti-freezing agent solves the problems of long reproductive cycle of female animals, low artificial insemination rate, complex production operation of frozen donkey semen and high difficulty in the prior art.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a casein antifreeze agent for donkey semen cryopreservation and a preparation method thereof. Background technique [0002] The combination of semen cryopreservation and artificial insemination technology plays a huge role in modern animal husbandry production. At the same time, semen preservation and utilization technology is also an important means to tap the reproductive potential of excellent male donkeys; the use of frozen semen can improve the utilization efficiency of breeding male donkeys and reduce the number of breeding male donkeys; Male donkeys have excellent production performance and genetic quality, which speeds up the genetic improvement of donkeys; artificial insemination technology can also reduce the occurrence and spread of reproductive tract diseases, and indirectly increase the conception rate of female donkeys; Without geographical restrictions, it is equivale...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 张燕王长法张瑞涛朱涛涛王玲玲鞠志花黄金明齐超高运东仲跻峰侯明海王银朝闫金华
Owner SHANDONG TIANLONG ANIMAL HUSBANDRY SCI & TECHNOLEGY
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