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A kind of horse semen cryopreservation solution and preparation method thereof

A cryopreservation solution and preparation method technology, which is applied in the field of equine semen cryopreservation solution and its preparation, can solve the problems such as incomplete clarity of change law and molecular mechanism, inconvenient on-site preparation and use, inconsistent composition of diluent and thawing solution, etc.

Active Publication Date: 2018-06-26
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the freezing technology of domestic horse semen is still in the experimental stage. The main reasons are: (1) the mechanism of freezing and thawing during the freezing process of horse semen, the mechanism of sperm damage, the impact of external environment changes on sperm, the change law of the internal components of sperm and Molecular mechanism and many other aspects are not completely clear; (2) The freezing-thawing procedure in actual operation, the composition of diluent and thawing liquid are inconsistent, and the composition and method are complicated, which is not convenient for on-site preparation and use in production; (3) Frozen semen It is also difficult for the vitality to meet the requirements of artificial insemination
The above problems seriously restrict the application of frozen horse semen in horse breeding and commercial production

Method used

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  • A kind of horse semen cryopreservation solution and preparation method thereof
  • A kind of horse semen cryopreservation solution and preparation method thereof
  • A kind of horse semen cryopreservation solution and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] A horse semen cryopreservation solution, consisting of 2.00g of glucose, 3.40g of lactose, 0.60g of trehalose, 0.061g of glutathione, 0.50g of sodium citrate, 30mL of skimmed milk, 2.5mL of egg yolk liquid, 50,000 IU of penicillin, strepto 50,000 IU of mycin, 3.5 mL of glycerol and double distilled water were used to prepare 100 mL of horse semen cryopreservation solution.

[0051] Concrete preparation method comprises the following steps:

[0052] (1) Preparation of skim milk: take fresh mare’s milk, filter it, sterilize it in a water bath at 100°C for 10 minutes, slowly cool it down to room temperature, put it in a refrigerator at 4°C, and then centrifuge it at 2000r / min for 30 minutes to remove the upper layer of lipid and the lower layer of sediment, get skim milk;

[0053] (2) Prepare base liquid I: Take glucose, lactose, trehalose, glutathione, sodium citrate, skimmed milk, egg yolk liquid, penicillin and streptomycin according to the amount, and first add glucos...

Embodiment 2

[0057] A horse semen cryopreservation solution, consisting of 2.40g of glucose, 4.00g of lactose, 1.00g of trehalose, 0.092g of glutathione, 0.70g of sodium citrate, 50mL of skimmed milk, 4.5mL of egg yolk liquid, 60,000 IU of penicillin, strepto 80,000 IU of mycin, 5.5 mL of glycerol and double distilled water were used to prepare 100 mL of horse semen cryopreservation solution.

[0058] The preparation method of the aforementioned horse semen cryopreservation solution specifically comprises the following steps:

[0059] (1) Prepare skim milk: add skim milk powder into double-distilled water, stir, and prepare a solution with a concentration of 0.1 g / mL to obtain skim milk;

[0060] (2) Prepare base liquid I: Take glucose, lactose, trehalose, glutathione, sodium citrate, skimmed milk, egg yolk liquid, penicillin and streptomycin according to the amount, and first add glucose, lactose, trehalose, glutathione Put glycine and sodium citrate into a sterile container, add double ...

Embodiment 3

[0064] Horse semen cryopreservation solution, consisting of 2.20g glucose, 3.70g lactose, 0.80g trehalose, 0.077g glutathione, 0.60g sodium citrate, 40mL skimmed milk, 3.5mL egg yolk liquid, 100,000 IU penicillin, streptomycin 100,000 IU, 4.5 mL of glycerol and double distilled water were used to prepare 100 mL of horse semen cryopreservation solution.

[0065] The preparation method of horse semen cryopreservation liquid specifically comprises the following steps:

[0066] (1) Preparation of skim milk: take fresh mare’s milk, filter it, sterilize it in a water bath at 100°C for 10 minutes, slowly cool it down to room temperature, put it in a refrigerator at 4°C, and then centrifuge it at 3000r / min for 20 minutes to remove the upper layer of lipid and the lower layer of sediment. get skim milk;

[0067] (2) Prepare base liquid I: Take glucose, lactose, trehalose, glutathione, sodium citrate, skimmed milk, egg yolk liquid, penicillin and streptomycin according to the amount, a...

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Abstract

The invention discloses a cryopreservation liquid for horse semen. 100 mL of the cryopreservation liquid for horse semen is prepared from 2.00-2.40 g of glucose, 3.40-4.00 g of lactose, 0.60-1.00 g of trehalose, 0.061-0.092 g of glutathione, 0.50-0.70 g of sodium citrate, 30-50 mL of skimmed milk, 2.5-4.5 mL of egg yolk, 50,000-100,000 IU of penicillin, 50,000-100,000 IU of streptomycin, 3.5-5.5 mL of glycerin and double distilled water. The invention further discloses a preparation method of the cryopreservation liquid. The cryopreservation liquid for horse semen can effectively increase the survival rate and semen activity of the horse semen during cryopreservation. The unfrozen semen has high physiological indexes, has the stable and reliable fecundation effect after field application and can be applied to production. The preparation method comprises few steps and is simple and easy to operate, and the preparation condition is mild and easy to control.

Description

technical field [0001] The invention relates to a horse semen cryopreservation solution and a preparation method thereof, belonging to the technical field of animal semen cryopreservation. Background technique [0002] Semen cryopreservation is one of the means of artificial assisted reproductive technology in horses. In 1950, Smith and Polge first applied glycerol as an antifreeze agent to cryopreservation of horse semen. In 1957, Barker and Gandier were the first to use frozen semen from the epididymis of horses for breeding conception. As early as the end of the 1950s, China began to conduct research on the cryopreservation of horse semen, and achieved good results. In the 1980s, as breed registration organizations such as Arabian horses, quarter horses, and American pinto horses allowed the registration of foals born from breeding with frozen semen, the cryopreservation technology of horse semen developed rapidly. Freezing horse semen can expand the breeding range of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/02
Inventor 阿娜尔白东义赵一萍房元海格力格桑芒来
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY