The method of processing the sperm of a bull

 

The invention relates to veterinary medicine. In lactose-glycerine-yolk medium injected 0.1-0.3% of chitosan succinate and obtained a synthetic medium is diluted with whole semen of a bull, after which it is subjected to equilibration and freezing in the vapor of liquid nitrogen. The method can reduce the biological damage to membrane structures of sperm in the process of deep freezing and increases their survivability after thawing. table 2.

The invention relates to veterinary medicine, in particular to methods of artificial insemination of animals, and in particular to methods of processing and freezing semen of a bull.

A method of processing the sperm of a bull, according to which frozen semen is diluted with a protective environment in diluted semen add decalin, has an antioxidant effect, is subjected to equilibrate and frozen in the vapor of liquid nitrogen.with. The USSR 1331499, class A 61 D 7/02, 1987). The method allows to stabilize the biological integrity of the sperm and reduce the cost of food - eggs for the purpose of artificial insemination of animals. However, the applied drug decalin roads and scarce.

A method of processing sperm selskohozyaistvennyi 1 billion sperm; the sperm was diluted protective environment, equilibrium and frozen in the vapor of liquid nitrogen to -196o(A.with. The USSR 1144698, class A 61 D 7/02, 1985). The method contributes to improving the security of membrane structures of sperm during freezing and thawing. However, this method, like the previous one, does not always contribute to improving the survivability Alannah sperm. Used a drug methyluracil also relatively expensive.

A method of processing the sperm of a bull, taken as a prototype, according to which their semen is diluted with a synthetic environment, including: lactose - 11.5g, the yolk of chicken eggs to 20.0 ml, glycerol 5 ml, spermon - 3-50 thousand units , distilled water - 100 ml Diluted semen cooled at 2-5oC for 3-4 hours, subjected to equilibrate and freeze in the form of granules with a volume of 0.2 ml in Teflon plate in pairs nitrogen and stored in nitrogen at -196o(Instructions on organization and technology work stations and enterprises artificial insemination of farm animals. - M.: Kolos. - 1981. - 159 S.). The method allows to obtain in one octanol spermatoza 10 million active sperms and use the available components, except starmoana-3. Cons the of membrane structures of sperm.

When creating the present invention, the objective was to increase the survivability of sperm after thawing, the expansion of the Arsenal used cryoprotectants (antioxidant agents) and the prevention of damage to membrane structures of spermatozoa during freezing sperm.

The technical result of the invention is achieved by a method for processing semen of the bull, including the dilution of whole sperm, lactose-glycerine-yolk medium, equilibration and freezing in pairs geckogo nitrogen, which feature is the introduction into the environment of 0.1-0.3% chitosan succinate.

Chitosan is a N-deacetylation form of chitin. It has antioxidant, sorption and antibacterial properties that contribute to the inhibition of the process of lipid peroxidation in the freezing of sperm and prevent damage to membrane structures of sperm substances generated during the peroxidation of lipids by sorbirovaniya. In addition, chitosan increases the viscosity createsite environment, which also contributes to reducing the cryogenic damage sperm.

According to the proposed method, the sperm of a bull is treated according to the following scheme. Their ejaculatory 0.1-0.3% of chitosan. Then the diluted sperm equilibrium 2-3 hours at +4oAnd frozen using a Teflon-coated plate in liquid nitrogen to -196oWith in the form of granules with a volume of 0.2 ml of the Thawed sperm pellets in a 3% solution of sodium citrate at +37oC.

Example 1.

In 2001, CSI was tested the effect of different dosages of chitosan on the stability of the sperm of a bull to deep freezing to -196oC.

Their ejaculates were diluted in synthetic medium to a final concentration of 30 million sperm cells in 0.2 ml With different dosages of chitosan was pre-dissolved in the following createsite environment: - distilled water - 100 ml; - lactose - 11.5g; glycerol 5 ml; yolk of eggs 20 ml.

The diluted semen was cooled in the refrigerator up to +4oC for 3 hours, and then froze in the form of granules with a volume of 0.2 ml in Teflon plate in pairs nitrogen and stored in nitrogen at -196oC. a Control sample of semen was diluted in a known environment (prototype), but without adding chitosan and froze in the same pattern. Thawed pellets sperm in 3% solution of sodium citrate and then incubated in an incubator at +37oC. Biological integrity of sperm, oily survivability of sperm.

Table 1 shows the biological integrity octanol sperm in the experimental and control groups.

From table 1 it is seen that the chitosan doses 3-0,12 mg/ml of the environment have had a positive impact on the viability of frozen-Alannah sperm of a bull. Optimal dosages of chitosan in the composition of the diluent is 1-3 mg of the drug per 1 ml of medium; thus the absolute measure of survivability Alannah sperm of a bull in groups 1, 2 and 3 mg of chitosan in 1 ml of medium was respectively higher than in the known way on 23, 24 and 22% (P<0,05). Lower concentrations of the drug had a less marked effect on increasing the survivability Alannah sperm. Sperm motility immediately after thawing was almost the same for all studied doses of chitosan.

Example 2.

Examined the influence of optimal dosages of chitosan on the stability of the sperm of a bull to the deep freeze, depending on the concentration of yolk of chicken eggs in the composition of the diluent.

Dilution and freezing of semen in this example was carried out similarly to example 1.

As can be seen from table 2, the introduction of chitosan in optimum doses of the composition of the freezing medium, stereoscopically chitosan had almost the same protective effect. Higher survivability of spermatozoa was observed in the group with 20% egg yolk in the composition of the diluent. In this group, with the introduction of part diluent 1, 2 and 3 mg of chitosan in 1 ml of medium, the viability of sperm was respectively higher than in the control 56, 48, and 41% (P<0,05).oAfter thawing and replace deficient antioxidant more affordable.

The invention can be used in the work stations and enterprises artificial insemination of cows and heifers.

Claims

The method of processing the semen of the bull, including the dilution of whole sperm, lactose-glycerine-yolk medium, equilibration and freezing in the vapor of liquid nitrogen, characterized in that the medium is injected 0.1-0.3% of chitosan succinate.

 

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