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Method for producing and thawing grain type frozen semen of pig

A granular and frozen semen technology is applied in the field of preparation and thawing of pig granular frozen semen, which can solve the problems of lower fertilization rate and litter size than fresh semen, complex operation procedures, and difficulty in popularization and application, and achieve simple formula, Simple operation and cost reduction effect

Inactive Publication Date: 2008-07-16
CHONGQING ACAD OF ANIMAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problems are: the composition of freezing liquid and thawing liquid is inconsistent, the amount of insemination is large, the fertilization rate and litter size are lower than fresh semen, the operation procedure is complicated, and it is difficult to popularize and apply in production

Method used

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  • Method for producing and thawing grain type frozen semen of pig
  • Method for producing and thawing grain type frozen semen of pig

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The preparation of embodiment 1 granular frozen semen

[0039] 1. Collection and routine inspection of pig semen: before semen collection, wipe the boar’s abdomen with 0.1% potassium permanganate solution, and collect the pig’s sperm-rich segment of semen by hand holding method and put it in a semen collection cup covered with four layers of sterile gauze . Take the collected pig semen back to the laboratory within 30 minutes, observe that the semen is milky white, cloudy, smells slightly fishy, ​​and the microscopic examination viability rate is above 0.7, which can be used for cryopreservation.

[0040] 2. Preparation of diluent: Glucose 2.0g, lactose 1.0g, sucrose 1.5g, glycine 0.8g, sodium citrate 0.3g, SDS 0.25g, caffeine 0.0971g, dissolved in 100ml distilled water as the base solution, penicillin and strepto Mycin each 1000IU / ml. Take 80ml of base solution and add 20ml of egg yolk to make liquid I, and add 3ml of glycerin to 100ml of liquid I to make liquid II. ...

Embodiment 2

[0043] Example 2 Thawing of frozen semen

[0044] 1. The formula of the thawing solution: 5.0 g of glucose, 0.5 g of sodium citrate, 5 ml of sodium citrate (10%), 100000 IU of penicillin and streptomycin, and 100 ml of distilled water.

[0045] 2. Thawing method: Take the frozen semen granules in the cloth bag out of the liquid nitrogen, stay in the air for 20 seconds, then quickly put them into the preheated thawing solution in a 42°C water bath, shake the test tube for 8 seconds to promote the full mixing of semen and thawing solution . The amount of thawing liquid to be added is 1ml of thawing liquid per frozen semen, and kept at room temperature for later use.

Embodiment 3

[0046] Example 3 Quality Assessment Method and Judgment Index of Frozen Semen

[0047] After the semen was thawed, its vitality, acrosome integrity rate, tail bending rate, deformity rate, recovery rate and survival time were detected to comprehensively evaluate its quality. The specific operation is as follows:

[0048] 1. Vitality detection

[0049] After the semen is thawed, place it at room temperature for 3-5 minutes, take 50ul of semen and drop it on a glass slide preheated at 42°C, quickly cover it with a cover glass, and observe the percentage of sperm moving in a straight line under a microscope. The viability rate after thawing is not less than 0.52.

[0050] 2. Acrosome Integrity Rate

[0051] The acrosome integrity rate of sperm after thawing was mainly evaluated by Coomassie brilliant blue staining. The specific operation is as follows:

[0052] (1) Preparation of solution

[0053] A. Weigh 50mg of Coomassie Brilliant Blue R250, dissolve it in 100ml of disti...

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Abstract

The invention relates to a method for preparing and unfreezing boar particle frozen semen. The steps of the method are of collecting and routinely examining boar semen, , preparing a diluent, diluting and balancing the semen, freezing and preserving the semen and unfreezing the frozen semen. The invention also relates to preparation of the diluent and a thawing solution. Compared with the prior art, the invention has the advantages that: after water-bath balancing, the fresh semen is directly centrifuged and no other diluents is needed to add, thereby reducing operation steps and reducing cost; the particle frozen semen is easy to operate and convenient to take; the thawing solution is easy to prepare and obtain and has the function of unfreezing and preserving the semen. The semen can survive at least for 420min after being unfrozen.

Description

technical field [0001] The invention relates to a method for preparing and thawing pig granule-type frozen semen. Background technique [0002] Semen cryopreservation plays a very important role in modern animal husbandry. The use of frozen semen can not be limited by time, region and lifespan of male animals, reduce the number of male animals, reduce production costs, improve the utilization rate of high-quality breeding animals, and bring huge economic benefits to animal husbandry production. At the same time, the cryopreservation of semen is also of great significance in breed improvement, blood renewal, disease control and protection of endangered animal breeds. [0003] Since the 1950s, the technology of freezing pig semen has made slow progress. In developed countries, the technology of freezing pig semen can basically be applied to the production of pig industry. However, from a global perspective, the application of pig semen technology is not ideal. This technolog...

Claims

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Application Information

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IPC IPC(8): A61D19/00
Inventor 王金勇蔡元潘红梅麻常胜白小青张凤鸣陈四清李琴刘文王可甜谷山林
Owner CHONGQING ACAD OF ANIMAL SCI
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