Semen diluent for improving cryopreservation quality of donkey semen, as well as preparation method and application thereof

A low-temperature preservation and diluent technology, which is applied in the field of animal husbandry, can solve the problems of sperm ultrastructural damage, low fertilization rate, long estrus time of donkeys, etc., and achieve the effect of improving the activity of low-temperature preservation of semen, broad market prospects, and prolonging the preservation time

Active Publication Date: 2018-05-04
CHINA AGRI UNIV +1
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  • Application Information

AI Technical Summary

Problems solved by technology

For example, the cryopreservation technology of bovine semen is relatively mature, especially the procedural production of bovine semen with thin tube method has been popularized; however, there are many problems such as unstable conception rate in estrus, low conception rate and low repetition rate of sheep sperm after freezing; and pig sperm Due to high lipid content, dehydration is difficult, ultrastructural damage of spermatozoa after freezing-thawing (such as sperm membrane, acrosome, and sperm nucleus vesicles, damage or rupture), shortened life span in the female reproductive tract, and low fertilization rate and other issues, so pig semen cryopreservation is still in the experimental stage
At present, there are few researches about the cryopreservation of donkey semen at home and abroad. The main reasons are: on the one hand, the reproductive rate of donkeys is low and the gestation period is long; The donkey itself is particularly sensitive to low-temperature blows, and is easily damaged during freezing and thawing, so the semen after refrigeration and freezing greatly reduces the conception rate; third, due to the long estrus time of donkeys, the inaccurate timing of artificial insemination causes the pregnancy rate Low; finally, the average ejaculation volume of male donkeys is as high as 70ml per ejaculation, the density is relatively thin, and the electrolyte components in semen can promote premature aging of sperm
Therefore, the production of frozen semen of donkeys is complicated and difficult, which seriously restricts the application of frozen semen

Method used

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  • Semen diluent for improving cryopreservation quality of donkey semen, as well as preparation method and application thereof
  • Semen diluent for improving cryopreservation quality of donkey semen, as well as preparation method and application thereof
  • Semen diluent for improving cryopreservation quality of donkey semen, as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0045] The semen diluent for improving the quality of semen cryopreservation comprises A solution, skimmed milk and fresh chicken yolk; the volume ratio of the A solution to the skimmed milk is 1:1. The specific composition of the semen dilution is shown in Table 1.

[0046] Table 1

[0047]

[0048] The semen of the same male donkey was preserved with the four dilutions numbered SM1-SM4 in Table 1 above; fresh semen was used as a positive control. Regularly detect the sperm motility of different storage time (12h, 24h, 48h, 72h, 96h, 120h). All the experimental results were repeated 6 times independently, and the statistical analysis was carried out by SPSS software.

[0049] Said method of preserving semen:

[0050] 1) Semen collection; the sperm motility of the semen is required to be over 90%;

[0051] 2) Centrifuge the qualified semen at low speed (about 600g) at room temperature (about 10min), discard the supernatant; add 4 times the volume of semen diluent (about...

experiment example 2

[0059] Experimental example 2 Effects of low temperature storage for different time on female donkey insemination

[0060] Breeding test: female donkey groups were randomly divided into groups; the fresh semen of male donkeys and the semen stored at 4°C for different periods of time using the diluent SM3 in Example 1 and INRA96 (Semen Preservation Solution of Casu company in France) were taken respectively. Randomly used for mating female donkeys in estrus. Test donkey estrus identification: Breeders pass B-ultrasound rectal examination every morning, and judge the mating time according to the follicular development of female donkeys. When the diameter of the follicle is greater than 35mm, insemination is performed at 5-6 pm on the same day, the volume of insemination is 15ml, and the total number of sperm is 1 billion. Check the follicles again in the morning of the second day after insemination. If the female donkey does not ovulate, then inseminate again in the afternoon o...

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Abstract

The invention relates to the field of animal husbandry, and particularly discloses a semen diluent for improving the cryopreservation quality of donkey semen, as well as a preparation method and application of the semen diluent. The semen diluent provided by the invention can significantly improve the cryopreservation activity of the semen and prolong the in-vitro preservation time of sperms. Theapplication provided by the invention is simple in operation, low in application cost, significant in effect and free of any harmful effect on animals. The semen diluent can be widely applied to donkey animal semen preservation and related embryo transfer industry and embryo engineering, and has a wide market prospect.

Description

technical field [0001] The invention relates to the field of animal husbandry, in particular to a semen diluent for improving the quality of semen cryopreservation and its preparation method and application. Background technique [0002] The combination of semen cryopreservation and artificial insemination technology plays a huge role in modern animal husbandry production. It can greatly reduce the number of male animals raised, reduce production costs, improve the utilization rate of excellent breeding male animals, and bring huge economic benefits to animal husbandry. benefit. At present, donkey breeding mainly adopts the method of self-breeding, and some skilled animal husbandry personnel sometimes inseminate the collected semen (fresh semen) without diluting or diluting it to the estrous female donkey immediately, which is suitable for seasonal estrus of female donkeys. More obvious and more numerous areas. Semen cryopreservation technology can make full use of the gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0221A01N1/0226
Inventor 曾申明吴帅帅杨峰于杰
Owner CHINA AGRI UNIV
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