Effect Of DMF、EG And Sugars On The Semen Cryopreservation Of Sika Deer

This study was conducted to optimize the traditional semen extender of sika deer and discuss theequilibrium time as well as thawing procedure. The sperm quality was evaluated by tests of spermmotility, movement characteristics, plasma membrane integrity, acrosome integrity. Results indicated asfollows:1. Three levels (4%,6%,8%) of glycerol were added to the traditional extender (Tris-citricacid-glucose-egg yolk-glycerol) to test the effect of glycerol levels on frozen sperm of sika deer.Significantly higher sperm motility (62.00%), progressive sperm percentage (44.22%), plasmamembrane integrity (40.00%), acrosome integrity (59.50%) and VCL (116.86μm/s) were obtainedusing6%GLY compared with other treatments (P<0.05), indicating6%GLY was the most suitablelevel for sika deer sperm freezing.2. Glycerol as a cryoprotectant was toxic to sperm, in recent years many researches have been done tolook for other cryoprotectants with lower toxicity. In this study DMF and EG were used to partly orfully replace glycerol. Significantly higher sperm motility (42.33%), plasma membrane integrity(39.83%), acrosome integrity (35.00%) and survival index (92.25) were obtained using4%EGcompared with6%GLY (P<0.05), indicating4%EG could be used to fully replace glycerol. DMFproved to be ineffective cause the parameters mentioned above were significantly lower than thoseof6%GLY treatment (P<0.01).3. Sugars can interact with other matters in extender leading to various protection efficiency. When theextender contains glycerol, egg yolk or Tris, glucose and fructose act differently. Fructose andsucrose were chose to replace glucose in the traditional extender aiming at analyzing the effect ofdifferent sugars in the Tris-citric acid-egg yolk-glycerol extender. Significantly higher spermmotility (49.50%), acrosome integrity (40.60%), VCL (143.30μm/s), ALH (6.98μm) and survivalindex (119.81) were obtained using fructose compared with other treatments (P<0.05).4.2h,3h and4h were chosen to explore the optimum equilibrium times in sika deer semencryopreservation. Significantly higher sperm motility (68.00%), sperm progressive percentage(38.33%), acrosome integrity (40.60%), VSL (73.35μm/s), VCL (159.27μm/s) and ALH (7.15μm)were obtained using4h compared with2h treatments (P<0.01), indicating4h was more suitablefor the cryopreservation.5. Four programs (Ⅰ37℃,20s; Ⅱ40℃,15s; Ⅲ50℃,10s; Ⅳ60℃,8s) were chosen to explorethe optimum thawing procedure in sika deer semen cryopreservation. Significantly higher spermmotility (50.33%), sperm progressive percentage (35.33%), plasma membrane integrity (33.37%),acrosome integrity (38.66%), VSL (80.33μm/s), VCL (151.60μm/s), ALH (6.33μm) and LIN(51.00%) were obtained using program Ⅰcompared with other treatments (P<0.05), indicatingprogram Ⅰwas the optimal option for the cryopreservation.