| Tarim rabbit is a unique species in China,which is in the national second class of preserved animals.In recent years,the damage to forestry and agriculture in some areas of South Xinjiang has increased year by year.However,in recent years,the domestic demand for hares has become more and more popular,and poaching and illegal reselling have occurred and are on the rise.In order to meet the demand of the market for the Tarim rabbit and reduce the stress of the wild population,and the protection and utilization are closely integrated by domestication and breeding.However,the wild rabbits in Tarim are strong,small and natural reproduction rate is low,and their reproductive mechanism is unknown,and the artificial propagation technology is still blank.The study on the collection and preservation methods of the rabbit sperm in Tarim Basin will provide experimental data for the establishment of the sperm bank and the establishment of artificial insemination technology system.In this study,taking Tarim rabbits as the test object,using modern animal reproductive technology,study the semen collection and preservation method of tarim rabbits,this study includes 6 parts.1.Screening of the species and dose of anesthesia type and the dosse suitable for Tarim rabbit : Three kinds of anesthesia drugs(Sumianxin?,3 % Pentobarbit and 20 % Urethane),according to different doses of anesthesia drug,the time required for anesthesia,anesthesia duration,anesthesia,adverse reactions and other indicators,to explore different effects of Anesthesia drugs and their Dosage on Anesthesia effect of Tarim Rabbit.The results showed that all the three narcotic drugs could effectively induce the anesthesia of Tarim rabbit,and the anesthesia effect of Sumianxin?(0.05 mL/kg)was better,the time required for anesthesia was 6.3 min,the duration of anesthesia was 47 min,followed by 3 % pentobarbital sodium(0.9 mL/kg),the time required for anesthesia was 7.3 min,the duration of anesthesia was 63 min,adverse reactions occurred,intramuscular injection would produce local anesthesia,and the rats were sacrificed.20 % Urethane(3.0 mL/kg),the time required for anesthesia was 5.0 min,the duration of anesthesia was 35 min,and adverse reactions occurred.Intramuscular injection was localized and intravenous.2.Screening method for artificial extraction of Tarim rabbit.The effects of different spermatogenesis methods on the semen collection of Tarim rabbits were studied by using different sperm methods(anatomical semen collection method,electric stimulation method and massage method)for semen collection,testing of semen volume,sperm motility and sperm density.The results showed that the three kinds of sperm extracts were able to collect semen,and the massage method had the best effect,and the fine amount was 0.20 mL,the sperm motility was average more than 0.65 and the operation was simple,The best ejaculation voltage of 0.1-3.0 V,the semen volume reach at 0.15 mL,sperm motility on average more than 0.45,but the operation of the tedious technical requirements are higher;compared with the anatomical semen collection method sperm motility average of 0.73 or more,but need slaughter the male rabbit.3.Screening for Tram rabbit semen preserves diluent.The rabbits semen were treated with different dilutions(physiological saline,dilution 1,2,3 and 4)at a 1:5 ratio and stored at room temperature(20-25 ?)at different times(0,4 and 8 h).The effects of different dilutions on the preservation of semen in Tarim rabbit were investigated by the method of continuous preservation.The results showed that spermatozoa were preserved in 5 dilutions,and the sperm motility and plasma membrane integrity decreased gradually(P<0.05).The sperm deformity rate increased(P<0.01),but there was no significant difference between the two groups(P>0.05),and there was no significant difference between the two groups(P>0.05),but there was no significant difference between the two groups(P<0.01)(37 h),with 3(30 h),with 4(28 h)(P<0.05),but with 3(37 h)And there was no significant difference between the two groups(P>0.05).4.Screening Tarim Rabbit Semen Preservative Dilution pH.The pH of the dilution(2),adjusted with the previous test,was adjusted to 6.2,7.2 and 8.2 have stored at room temperature(20-25 ?)and sampled at different time points(0,4 and 8 h)sperm motility,deformity rate and plasma membrane integrity.The effect of dilution pH on semen preservation efficiency of Tarim rabbit was studied by the method of continuous preservation.The results showed that the spermatozoa of Tarim rabbits were preserved in different pH values,and the sperm motility and plasma membrane integrity decreased gradually(P<0.05).The sperm deformity rate gradually increased(P<0.01).In the pH 7.2 group,the sperm survival time was the longest(42 h),and the sperm viability was the highest(P<0.01)significantly higher than pH 6.2(2.5 h),pH 8.2(6 h)(P<0.01).5.Screening method for preserving Traim rabbits.The dilution(2,pH 7.2),which was screened by the previous test,have stored at under the different temperatures.For example,low temperature(4-5?),room temperature(20-25 ?),constant temperature(37 ?)(0,4 and 8 h),the sperm motility,the rate of deformity and the integrity of plasma membrane was measured.The survival time of sperm was tested by continuous preservation method.The effects of different preservation methods on the preservation of semen in Tarim rabbit were discussed.The study results showed that the spermatozoa of Tarim rabbits were preserved under different preservation conditions,and the sperm motility and plasma membrane integrity decreased gradually(P<0.05),and the sperm deformity rate increased gradually(P<0.05)(0.81,0.78 compare to 0.45;0.71,0.76 compare to 0.31,respactivly)(P< 0.01),but there was no significant difference between low temperature and normal temperature group(P>0.05),but there was no significant difference between the two groups(P>0.05).The survival time of the spermatozoa was significantly higher than that of the cryopreserved group(P<0.05),which was significantly higher than that in the cryoprotective group(P<0.01).6.Evaluation of artificial insemination of semen preservation dilution.Tarim rabbit semen,respectively,in 5 kinds of semen preservation commonly used dilution(physiological saline,dilution 1,2,3 and 4)at room temperature(20-25 ?)preserved 2 h after artificial insemination,and compared with the natural mating,the effects of different dilutions on the fertilization and calving of Tarim rabbits were investigated by observing the conception rate and average litter size.The results showed that the conception rate of natural mating was 60 %,which was higher than that of other groups(P<0.05).The conception rates of normal saline,dilution 1,2,3 and 4 were 20 %,30 % and 40 %,30 %,30 %,of which dilution 2 of the fetus effect is higher to 40 %. |
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