| With the acceleration of social aging,one-child family work pressure and other objective factors prominent,people’s emotional sustenance has changed significantly,and pet keeping has become a way of entertainment for many families.The pet industry has developed gradually.The number of pet dogs in China is significantly higher than that of other pets,and now the demand is increasing.In recent years,with the demand of the market for the offspring of excellent breed dogs,the dog breeding industry has begun to develop professionally and scientifically,in which the technology of low temperature storage of dog semen and artificial insemination technology plays an important role in promoting the development of the industry.In order to make full use of the utilization rate of excellent breed dogs and meet the needs of the society for pet dogs,it is an urgent problem to dilute the semen with semen diluent and preserve it at low temperature,and finally obtain the offspring of excellent pet dogs by artificial insemination.At present,most of the diluents in common use must be added with yolk as a protective agent.However,the use of yolk is difficult to operate,which has the risk of carrying pathogenic microorganisms.The components of yolk are complex,and even some components can react with enzymes in sperm to produce peroxides,Therefore,the commercial use space of sperm diluent using yolk as protective agent is very limited.In the present study,soybean lecithin of different concentrations was added to the semen diluent to store dog sperm,and egg yolk was used the control,we compared the sperm motility,plasma membrane integrity rate,acrosome integrity rate and conception rate after artificial insemination.In addition,the serum progesterone level in the whole oestrus period of female dogs was detected,and the change rule of progesterone in oestrus period was summarized,The artificial insemination test was carried out to evaluate the conception rate at different concentrations of progesterone.The technology of artificial insemination was optimized,which laid a foundation for the promotion of dog assisted reproductive technology.The main results of this study are as follows:1.The semen of French Bulldog was chilled for 168 h with different concentrations of soybean lecithin(0.5%,1.0%,2.0%),the results showed that the sperm viability,motility,plasma membrane integrity and acrosome integrity in the 1.0% soybean lecithin group were significantly higher than those of the other two groups(P < 0.05)and significantly higher than that of the 20% egg yolk control group.2.The serum progesterone value of 57 French Bulldogs showing natural estrus was measured by enzyme-linked fluorescence.The results showed that the progesterone level increased slowly from day 0 to day 5;on the 5th day of estrus,the serum progesterone value appeared inflection point at 3.31 ± 0.90 ng/m L,and then increased exponentially;on the 6th day,the progesterone value was 6.58 ± 1.44 ng/m L;on the 12 th day of estrus,the level of serum progesterone reached 23.89 ± 2.93 ng/m L.Therefore,the level of serum progesterone in estrous dogs has been rising.3.Artificial insemination was carried out at different range of serum progesterone values during estrus of French Bulldog.The results showed that when the serum progesterone value was 20.01-25.00 ng/m L,the pregnancy rate was 63.1%,and the average litter size was 5.5;when the serum progesterone value was 15.01-20.00 ng/m L,the pregnancy rate was 47.3%,and the average litter size was 4.7;when the serum progesterone value was 10.00-15.00 ng/m L,the pregnancy rate was 36.8%,and the average litter size was 3.7.The results showed that the effect of artificial insemination was the best when the concentration of serum progesterone was 20.01-25.00 ng/m L.4.Forty eight French Bulldogs with a serum progesterone value of 20.01-25.00 ng/m L were inseminated artificially using sperm chilled for 48 hours.The results showed that the pregnancy rate in the 1.0% soybean lecithin group was 70.8%(17/24),and the total number of litter was 98.The pregnancy rate in the 20.0% egg yolk group was 62.5%(15/24),and the total number of pups was 80.The results showed that 1.0% soybean lecithin diluent was better than 20% egg yolk diluent in cryopreservation of dogs’ sperm for artificial insemination.Therefore,it is feasible to use 1.0% soybean lecithin as a protective agent instead of 20% egg yolk when the French Bulldog semen is preserved at 4℃;when the level of serum progesterone is in the range of 20.01-25.00 ng/m L,the effect of artificial insemination is significantly better than that of other serum progesterone values. |
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