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Freezing diluent for seminal fluid of livestock

A technology for freezing diluents and semen, applied in animal husbandry, preservation and application of human or animal bodies, etc., can solve problems such as reducing the sedimentation speed of sperm, and achieve the effects of inhibiting the formation of ice crystals, improving quality, and ensuring purity

Active Publication Date: 2016-06-08
YUNNAN ANIMAL SCI & VETERINARY INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the semen is stored at room temperature or 0-4 ℃ low temperature, it mainly reduces the energy consumption by inhibiting the sperm activity, so that the sperm can maintain the fertilization ability in a static state, and it can be used within 1-7 days, expanding the use of excellent male semen Scope, the use of polyvinyl alcohol in these techniques is mainly used as a film-forming agent to protect the membrane activity of sperm cells, and at the same time can reduce the sedimentation velocity of sperm, and play a role in protecting sperm cells when diluted at room temperature and stored at 0-4°C. , but there is no research report on the use of polyvinyl alcohol as an ice crystal inhibitor for cryopreservation of sperm

Method used

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  • Freezing diluent for seminal fluid of livestock
  • Freezing diluent for seminal fluid of livestock

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The semen of Yunnan semi-fine wool sheep was collected by electrical stimulation, and then the semen quality was tested. The indicators of sheep semen used for cryopreservation must meet: the motility is greater than 75%, and the sperm density is higher than 1×10 9 / ml, the deformity rate is less than 10%, and the semen volume is 1-2ml.

[0026] Prepare semen frozen diluent, the steps are as follows: respectively weigh 2.71 g of Tris, 1.4 g of citric acid, 1.0 g of glucose, 5 ml of glycerin, 100,000 IU of penicillin, 100,000 IU of streptomycin, and 0.1 g of polyvinyl alcohol. g, dissolve in ultra-pure water, stir well, add 10ml of fresh egg yolk inactivated at 56°C for 30min before use, mix well and set the volume to 100ml, centrifuge at 15000 rpm for 1 hour at 4°C, take the supernatant The solution was filtered through a 0.45 μm disposable filter for later use.

[0027] Mix the sheep semen and frozen diluent that meet the freezing requirements evenly in a ratio of 1:...

Embodiment 2

[0030] Nubian goat semen was collected using a false vagina, and then the semen quality was tested. Semen indicators for cryopreservation must meet: motility greater than 75%, sperm density higher than 1×10 9 / ml, the deformity rate is less than 10%, and the semen volume is 0.75-2ml.

[0031] Prepare semen cleaning fluid, its step is, take by weighing 2.71g tris hydroxymethyl aminomethane, citric acid 1.4g, fructose 1.0g, penicillin 100,000 IU, streptomycin 100,000 IU, polyvinyl alcohol 0.2g, dissolve in Ultrapure water, after mixing evenly, dilute to 100ml, filter with a 0.22μm disposable filter for later use.

[0032]Prepare semen frozen diluent by weighing 2.71 g of Tris, 1.4 g of citric acid, 1.0 g of fructose, 7 ml of glycerin, 100,000 IU of penicillin, 100,000 IU of streptomycin, and 0.2 g of polyvinyl alcohol. g, dissolve in ultra-pure water, stir well, add 15ml of fresh egg yolk inactivated at 56°C for 30min before use, mix well and set the volume to 100ml, centrifug...

Embodiment 3

[0036] The semen of the hornless Dorset ram was collected by the false vagina method, and then the semen quality was tested immediately. The indicators of sheep semen used for cryopreservation must meet: the motility is greater than 75%, and the sperm density is higher than 1×10 9 / ml, the amount of semen is 1-2ml.

[0037] Prepare a frozen diluent that does not contain an osmotic protectant. The preparation method is to weigh 2.71 g of Tris, 1.4 g of citric acid, 1.0 g of glucose, 100,000 IU of penicillin, 100,000 IU of streptomycin, and Polyvinyl alcohol 1.0g, dissolved in ultrapure water, stirred evenly, before use, add 20ml of fresh egg yolk inactivated at 56°C for 30min, mix evenly, dilute to 100ml, centrifuge at 15,000 rpm at 4°C for 1 hour , take the supernatant and filter it with a 0.45 μm disposable filter for later use.

[0038] Prepare semen frozen diluent. The steps are as follows: weigh 2.71g of Tris, 1.4g of citric acid, 1.0g of fructose, 10ml of glycerin, 100,...

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Abstract

The invention relates to a freezing diluent for a seminal fluid of livestock. Every 100 ml of the freezing diluent comprises components as follows: 2.71 g of trihydroxymethyl aminomethane, 1.4 g of citric acid, 1.0 g of monosaccharide, 5-20 ml of fresh yolk, 0-10 ml of a permeable protecting agent, 0.1 million IU of penicillin, 0.1 million IU of streptomycin, 0.05-5 g of polyvinyl alcohol and the balance of ultrapure water. The common high-molecular polymer polyvinyl alcohol serving as an ice crystal inhibitor replaces natural antifreeze protein and is applied to the cryopreservation research of the seminal fluid of livestock for the first time. The survival rate of unfrozen sperms is about 75%, the activity is 50% or higher, the acrosome completeness is 65% or higher, the plasma membrane completeness is about 50%, and the non-return rate after artificial insemination is 70% or higher.

Description

technical field [0001] The invention relates to the technical field of animal reproductive physiology and reproduction, in particular to a frozen diluent of livestock semen. Background technique [0002] Although the research on cryopreservation of livestock semen has been carried out for more than 50 years, the viability of thawed semen is only about 30%-50%, and the non-return rate and litter rate after artificial insemination are significantly lower than that of fresh semen. During the freezing process of livestock semen, the mechanical damage caused by the ice crystals formed inside and outside the cells directly leads to the decline of sperm motility after freezing. How to reduce the damage to livestock sperm produced by ice crystals has always been a research field in the field of livestock breeding technology and cryobiology. Hotspot, by changing the composition of semen diluent, the damage of freezing to sperm can be reduced. At present, osmotic organic solvents and ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/02
Inventor 权国波洪琼花邵庆勇吴国权吕春荣
Owner YUNNAN ANIMAL SCI & VETERINARY INST
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