猪精液颗粒冷冻技术的研究

Study on the pellet freezing technique of boar semen

为提高猪冻精质量,加速猪冻精在生产上的应用,本试验以解冻后的精子活率、顶体完整率和体外受精(IVF)结果作为判定指标,探讨了冷源和Equex.STM对猪精液颗粒冷冻效果的影响。手握法采集成年、健康种公猪精液,分别采用含有Equex.STM的稀释液和普通稀释液进行两步法稀释,然后以干冰或液氮为冷源进行精液冷冻。结果表明,以干冰为冷源滴冻精液冻后活率为0.487833±0.046761,顶体完整率为0.782±0.039704,而以液氮为冷源的方法冻后活率为0.3334±0.055734、顶体完整率为0.5578±0.068482,前者在这两项指标上均显著优于后者(P<0.05)。另外,在稀释液中添加Equex.STM,以干冰为冷源进行滴冻,可使冻精顶体完整率极显著提高(P<0.01)。以含有Equex.STM的稀释液采用干冰滴冻的冻精进行IVF,胚胎卵裂率、囊胚率分别为0.5316±0.125582和0.216±0.053198。结果表明,采用Equex.STM作为冷冻保护剂并以干冰作为冷源可以获得高质量的猪冷冻精子。

In order to improve the thawed sperm quality and make the cryopreserved sperm available in swine industry, the effects of Equex.STM and different freezing system on porcine sperm cryopreservation were studied. Thawed sperm quality was evaluated by motility, acrosome integrity and in vitro fertilization（IVF） rate. Ejaculated semen was collected by the gloved-hand method from healthy adult boars. Sperms were diluted by exterder with or without Equex.STM in two steps, and were frozen by dropping on dry ice or on copper mesh which were cooled by liquid nitrogen. The thawed motility and acrosome integrity of sperms frozen by dry ice or by liquid nitrogen was 0.487833±0.046761, 0.782±0.039704 or 0.3334±0.055734 and 0.5578±0.068482, and both the thawed motility and acrosome integrity of sperm frozen by dry ice was significantly higher （P〈0.05）than the sperms frozen by liquid nitrogen. Extender with Equex.STM and froze by dry ice improved the acrosome integrity significantly （P〈0.01） Thawed sperms from exterder with Equex.STM and frozen by dry ice were used to do IVF, and the cleavage rate and blastocyst rate was 0.5316±0.125582 and 0.216±0.053198 re-spectively. These results indicated Equex.STM and dry ice freezing system can provide high quality frozen sperm.