滇南小耳猪精液的直接冷冻保存

目的建立滇南小耳猪精液冷冻保存方法,加速云南省特有小型猪种的小型化选育。方法利用脉冲电刺激模式诱导公猪输精管自助收缩排精。利用直接冷冻新技术（DFM）研究不同冷冻方案对精子的运动度、精子顶体完整性和体内授精胚胎发育能力的影响。结果在直接冷冻方法中,3%甘油防冻剂的作用下,60 s植冰时间和1.5 mm/s的冷冻降温参数对精子运动度保护良好,精子运动复苏率达到61.7%。但是,3%乙二醇虽然与甘油一样对精子的顶体完整性都有很好的保护作用,对精子运动度保护能力较差。此外,3%甘油、60 s植冰、1.5mm/s冷冻速度的直接冷冻的冻精解冻,移植到超数排卵的母猪子宫颈口实施人工授精,获得卵的受精和胚胎发育潜能尚可。结论玻璃管直接冷冻可以完成滇南小耳猪精子的冷冻保存。

Cryopreservation of Sperms with Directional Freezing Method in Yunnan Diannan Miniature Pig

Objective To establish the cryopreservation of sperm genetic source and to speed up the selection of a special-strain minipigs.Methods Boars anesthetized were preformed rectums electroejaculation with Lane Pulsator IV machine.The freeing method of sperms was directional freezing method（DFM） with different seeding ice duration（60 s） and different speeds of cooling（0.5 ～ 2.5 mm /s）.Results In cryopreservation with direct freezing method（DFM）,the parameters of 60 s seeding ice duration and 1.5 mm /s cooling speed produced the better protection than others in the terms of the motility（61.7%） and acrosamal integrity of post thawed sperms.However,3% EG showed the deficient in potential of cryopreservation,though the acrosomal intergrity showed no difference with fresh sperms.The development potential of pronuclear embryos produced by the artificial insemination of frozen sperms with 60 s seeding duration and 1.5 mm /s cooling speed was acceptable.Conclusion Directional freezing method can perform sperm cryopreservation and fertilization of ova in vivo and sequent embryos can develop to blastocyst stage.