| We conducted the experiments to investigate the quality of ram semen which are feeded in changchun city dehui tiantai. Ten semen extenders were studied on the mobility of ram spermatozoa at three different temperatures in order to find out which is the best extender: the temperatures such as at roomtemperature(20-25℃), low temperature (0-5℃) and 39℃. This study also investigated optimum cryopreservation procedure, extenders adding soy bean phospholipids and the cervical insemination using the best extender. The results showed that:The quality of five ram's spermatozoa accord with criterion of reproducing request. The result indicates that the five rams are capable of hybidization, the number of the rams is 76, 121 , 748, 179 and 2056. The quality of the two other ram's spermatozoa is worse than the previous five ram's, the number of the ram is 1084 and 275. The two rams should reinforce breeding manage and improve the quality of ram in order to reach to reproducing criterion.The mobility of the ram's spermatozoa which conserve in Tris diluent is better than other diluents at room temperature,low temperature and 39℃(P<0.05). These results showed that Tris could be used to conserve ram semen at three temperatures .The equilibration rate of 0.21℃/min at 37℃ to 5℃ for 3h is significant better than other methods for frozen-thawed ram semen (P<0.05.35℃ thawing temperature for frozed-thawed semen mobility and intact acrosome rate were significant better than 45 ℃, 55℃ and 65℃ thawing temperature for frozed-thawed semen's (P<0.05) . When the thawing temperature was higher than 45℃, the frozed-thawed semen mobility and intact acrosome rate would dramaticly reduce (P<0.05) . 35℃ thawing temperature with VB12 was simple and effective.This study testified that the optimum glycerol concentration in pellet freezing diluents was 6.0%, in which frozed-thawed semen mobility and intact acrosome rate were significant better than other glycerol concentrations (P<0.05) . When glycerol concentration was higher than 6%, the frozed-thawed semen mobility and intact acrosome rate would begin to drop and this phenomena showed that if glycerol concentration exceeded 6% would injure to semen.The ram spermatozoa optimum cryopreservation procedure was established evaluated by some targets such as frozen-thawed semen motility and intact acrosome rate.①Dilutionrate is 1:3 of semenrextender; ?Semen diluent with two-step dilution method, the semen was diluted with basic diluent ,then descent temperature to 5°C adding to glycerol; (3) Equilibration method is 200ml water under 37°C for 3h; ?Semen pellet thawed at 35°C with vitamin B12 wet thawing.Tris diluent adding to 3.65% or 7.3% soy bean phospholipids had better effect on the quality of frozed-thawed semen, which had prominence difference compared with other soy bean phospholipids concentration (PO.05) .The ram semen intact acrosome rate was best when Tris diluent contain 7.3% soy bean phospholipids (PO.05) .The frozed-thawed semen motility and intact acrosome rate would begin to quickly decline when soy bean phospholipids concentration exceeded 7.3%. So we can draw the conclusion that Tris diluent containing 7.3% phospholipids is the optimum concentration.The effect of different temperatures used during solubilization of soy bean phospholipids has different impact on the quality of frozed-thawed ram semen. The frozed-thawed semen motility and intact acrosome rate were best when the dissolving temperature is 60 "C.Utilizing the best freezing extender adding soy bean phosphjolipids (Tris+7.3%soy bean phospholipid+6%glycerol) and freezing extender adding egg yelk (Tris +15%egg yolk +6%glycerol) to carry out cervical insemination, pregnancy rate are 59.5% and 52.7% respectively. The result indicated that egg yolk could be substituted for soy bean phospholipids. |
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