| Using hand-holding method to collect the York boar semen in this test,gather the middle part of abundent sperm,confecting different diluents , pre-treating it with different conditions , thawing with different liquids and ways, after that to inseminate;at the same time , there are also investigating the microbe contents and sorts and the learning low-temperature preservation of boar semen. The results were following: 1.In seven diluents,the liquids of No.5 and No.6 were better than others (P<0.05),the after freezing mobility were 0.46,0.50(pellet) and 0.40,0.49(tubule), recovery percent were 76.7%,74.6%(pellet) and 70.5%,78.3%(tubule), the difference between No.5 and No.6 was not notability(P>0.05);in same diluents ,there were no difference between pellet freezing-sperm and tubule freezing-sperm(P>0.05)。The ideal frozen temperature curve and the relation curve of beginning temperature and after-freezing mobility in pellet freezing-sperm were set down with different beginning frozen temperature , balance temperature ,and enter nitrogen temperature. 2.The pretreatment before freezing of setting different time,different ratio and time of centrifugal,different remnant of liquid on boar novelty semen were made, compared the effect of sperm freezing result on the different pretreatment. The result is following:the result before centrifuging of setting 60min in 37℃ (0.5ml tubule after-sperm motility 0.50 ,recovery percent 75.8%, 0.5ml pellet after-sperm motility 0.48 , recovery percent 72.5%,0.1ml pellet after sperm motility 0.47,recovery percent 70.6%)was better than 30min and 90min(P<0.05);the centrifuging ratio of 1000r/min for 10min (0.5ml tubule after-sperm motility 0.48 ,recovery percent 76.8%, 0.5ml pellet after-sperm motility 0.46 , recovery percent 73.5%,0.1ml pellet after sperm motility 0.44,recovery percent 69.4%)is better than 500r/min and 1500r/min for 7min and 13min(P<0.05);it is the best that the remnant of liquid is 1/1 than 1/2 and 0/1(P<0.05)。 3.Compared with different cryoprotectant (Glycerol,DMSO,Glycol,Tris,OEP) and different thaw-liquids design by ourselves and thawing methods. The results showed:Glycerol is the best cryoprotectant (P<0.05) and the best content is 3% (sperm motility 0.48)(p<0.05);the sequence of toxicity of different cryoprotectant is :Glycol> DMSO> OEP> Tris> Glycerol;the effect of the No.4 and the sperm liquids is better than others in evidence(Pbut there is no distinctive diversity(P>.05);it is the better of thawing methods with 75℃,4s(motility 0.47,acorosomal integrity 55.6%,sperm abnormality 18.32%)。 4. Using pellet and tubule sperm to inseminate test that freezing diluents and thaw liquids made by ourselves. The result showed :pellet and tubule sperm hadn't distinctive difference in effective living time and after thawn sperm(P>0.05);but the pellet sperm was better than tubule in percent fecundation and equal litters , the pellet sperm was 81.81%,85.71%,9.22 and 9.33,tubule sperm was75.00%,80.00%,8.17 and 8.25。 5.The average number of microbe in pre-semen and after balance were 1.5×104 /ml,1.1×104/ml;the average number of microbe in pellet freezing sperm (0.1ml,0.5ml) were 920 and 3750 , in tubule freezing sperm the number (0.25ml,0.5ml) were 1060 and 1520.The sorts of microbe had Grape,Intestines bacillus,Streptococcus and so on. 6.It was persevered by different extender with different ratio of low temperature preservation in 0℃～5℃.The result showed: the No. 2 could persevere 60±4.05h effective , there was very nobility difference with the others (P<0.01);The centrifuging ratio was better with 1000r/min and could persevere 50±3.68h effective (P<0.05)in low temperature. The ratio of diluents was better with 1:1(semen/diluents). |
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