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精液平衡、冷冻及解冻是冻精制作过程中三个必不可少的环节,对精液冷冻效果起着决定性作用。在马(Equus caballus)精子冷冻中针对这三个过程的研究较少,为进一步优化马精液冷冻方法,提高精液冻后质量,本研究比较不同平衡时间、冷冻方法及解冻程序对冻融后精子运动参数、质膜完整性及线粒体膜电势的影响。平衡120 min、180 min和240 min组冻融后精液活力及质膜完整性明显高于平衡0 min、45 min、90 min及8 h平衡组;距离液氮面2 cm和4 cm高度熏蒸冷冻获得了与程序冷冻仪冷冻法相似的冷冻效果;采用高温瞬时解冻法(75℃7 s和46℃20 s)比常规方法(37℃30 s)获得了更高的冻后精液活力(P<0.05)。综合上述结果,在马精液冷冻过程中综合采用120~240 min平衡,距离液氮面2~4 cm熏蒸法和高温瞬时解冻法(75℃7s和46℃20 s)可获得更好的精液冷冻效果。
Semen balance, freezing and thawing are three essential steps in the sperm production process, which plays a decisive role in the sperm freezing effect. In order to further optimize the equine caballus sperm freezing process for the three processes, in order to further optimize the horse sperm freezing method and improve the quality of sperm after freezing, this study compared the different equilibration time, freezing method and thawing program on frozen-thawed sperm Exercise parameters, plasma membrane integrity and mitochondrial membrane potential. After freeze-thaw cycles for 120 min, 180 min and 240 min, the semen vitality and plasma membrane integrity were significantly higher than those in the equilibrium group at 0 min, 45 min, 90 min and 8 h, and were fumigated at 2 cm and 4 cm from liquid nitrogen The freezing effect was similar to the freezing method of the program freezing system. The higher frozen thawed semen vitality was obtained by the high temperature instant thawing method (75 ℃ for 7 s and 46 ℃ for 20 s) than the conventional method (37 ℃ for 30 s) (P < 0.05). Based on the above results, a better sperm freezing can be obtained by using 120 ~ 240 min equilibration during equine freezing, 2 ~ 4 cm fumigation from liquid nitrogen and instantaneous thawing at high temperature (75 ℃ for 7s and 46 ℃ for 20s) effect.