应用生殖免疫方法制作性别化冷冻精液对奶牛性别控制的研究

Study on the Reproductive Immunization on Sexing Control in Dairy Cattle with Frozen Sexed Semen

本实验在种公牛站精液生产过程中应用生殖免疫技术方法 ,制作性别化冷冻精液 ,结合人工授精技术进行奶牛性别控制的研究实验。根据精子DNA含量存在的差异 ,利用荧光染料与精子DNA结合 ,通过蔗糖溶液密度梯度离心法 ,分离出和鉴别出牛精液中的X与Y精子作为抗原。经与小鼠免疫后 ,制成H Y抗血清IgG ,再经酶联免疫吸附法 (ELISA)析测表明 ,获得了具有一定纯度和工作效价的阳性抗Y精子的H Y抗血清IgG ,H Y抗血清对性别化精液冷冻前 ,解冻后活力的影响与对照组无显著差异。配种受胎试验结果表明 ,性别化冷冻精液在获得与正常冷冻精液相似的情期受胎率的同时 ,对后代性别比率有显著影响 ,奶牛产母犊率可达 60 7% ,比自然产母犊性比率理论值提高 1 0 7个百分点 (P <0 0 5 )。

This trial was proceeded in process of semen production in the Pure bred Sire Station to prepare the Frozen Sexd Semen by use of Reproductive Immunization,and was being realization of Sexing Control in Dairy Cattle in combination with the artificial insemination.Its action principle,features and application were described chiefly.It was found that it is possible for separation of X and Y chromosome bearing sperm using Sucrose Solution Density Gradient Centrifugation based on DNA difference,and it was also an evidence of using Fluorescent staining for inspectation of the X and Y chromosome bearing sperm. H-Y antigen was isolated from inbred mice and subsequently used to immunize female rats for preparing H-Y antiserum IgG.It was also an evidence that using H-Y antiserum IgG showed marked specific inhibition on Y chromosome bearing sperm. Results also indicated that they could be reached the similar conception rate in estrus with the Frozen Sexed Semen just as using with the normal frozen semen simultaneously,but a marked variation of sexuality at birth between them.A ratio of more than 60.7% female progeny in cows was successfully reached after artificial insemination with Frozen Sexed Semen pretreated with H-Y antiserum IgG in a big group of 117 cows in this trial.;