摘要
为了提高纯种西门塔尔牛冷冻胚胎分割后的半胚发育率,试验选取牛一级冷冻胚胎在胚胎保存液、mCR1aa(前期)液、mCR1aa(后期)液3组培养液中培养24 h,测定胚胎发育率;牛一级冷冻胚胎解冻后分为0,15,30,45,60,90,120 min组培养后进行分割,将对照组整胚和分割后的半胚培养24 h,测定胚胎发育率;牛一级冷冻胚胎分割后的半胚分为不经短暂培养(no short culture,NSC)和30 min组作为试验组,整胚分为NSC对照、30 min对照组,将4组胚胎分别装入含胚胎冷冻液的0.25 m L细管中,在4℃冰箱放置30 min,之后在室温(25℃)下推入胚胎保存液中洗3次,再培养24 h,测定各组胚胎发育率。将纯种西门塔尔牛一级冷冻胚胎利用优化后的分割条件进行二分操作,分为整胚、双半胚、单半胚,用胚胎移植枪直接移植到受体牛子宫角,并用双半胚进行同侧移植,测定受体母牛妊娠率、流产率、双犊率、犊牛成活率。结果表明:mCR1aa(前期)液组、mCR1aa(后期)液组胚胎发育率极显著高于胚胎保存液组(P<0.01);试验组中解冻后培养45 min组胚胎发育率最高,与对照组无显著差异(P>0.05);NSC组胚胎发育率极显著高于30 min组(P<0.01);整胚、双半胚、单半胚组的妊娠率、流产率、犊牛成活率差异不显著(P>0.05),双半胚组的双犊率极显著高于整胚组、单半胚组(P<0.01)。说明纯种西门塔尔牛冷冻胚胎解冻后用mCR1aa(后期)液培养45 min再分割,获得的半胚直接移植可有效提高冷冻胚胎利用效率和分割后半胚移植妊娠率。
In the experiment,the first-grade frozen embryos of cattle were cultured in three groups of embryo preservation solution,mCR1 aa(early stage)and mCR1 aa(late stage)for 24 h,and the embryonic development rate of each group was determined.After thawing,the first-grade frozen embryos of cattle were divided into 7 test groups,which were 0,15,30,45,60,90,120 min group.Test groups were cultured and then split.The whole embryos of control group and the demi-embryos of splitting were cultured for 24 h,and the embryonic development rate of each group was determined.The demi-embryos of the first grade frozen embryos of cattle were divided into two test groups of no short Culture(NSC)and 30 minutes.The whole embryos were divided into NSC control group and 30 minutes control group.The embryos of four groups were put into thin tubes of 0.25 mL which contained embryo frozen solution.Then they were placed in a refrigerator at 4℃for 30 minutes.Later they were washed 3 times in the embryo preservation solution at room temperature(25℃).Finally they were cultured for 24 h.The embryonic development rate of each group was determined.Using the optimized split condition for the first-grade frozen embryos of purebred Simmental cattle are divided into three groups which were whole embryo,double demi-embryo and single demi-embryo.The embryos of three groups were directly transplanted to the recipient’s uterine horns with embryo transfer guns.The double demi-embryos were transplanted on the same uterine side.The pregnancy rate,miscarriage rate,double calves rate,and calves survival rate of the recipients were determined.The results showed that the embryonic development rate of the mCR1 aa(early stage)group and mCR1aa(late stage)group was significantly higher than that of the embryo pre-servation solution group(P<0.01);the embryonic development rate of 45 minutes group was the highest,and there was no significant difference in embryonic development rate between 45 minutes group and control group(P>0.05);the embryonic development rate of the NSC group was significantly higher than that of the 30 minutes group(P<0.01);the pregnancy rate,abortion rate and calves survival rate of the three groups of whole embryo,double demi-embryo and single demi-embryo were not significantly different(P>0.05).The double calves rate of the double demi-embryo group was significantly higher than that of the whole embryo group and single demi-embryo group(P<0.01).In conclusion,after thawing frozen embryos of purebred Simmental cattle,they were cultured in mCR1aa(late stage)for 45 minutes and then split,the demi-embryos which were directly transfered could effectively improve the utilization efficiency of frozen embryos and the pregnancy rate of demi-embryo transfer after splitting.
作者
赵增元
郭伟婷
毕江华
冯春涛
朱宏波
张普
孙建辉
王敏生
余文莉
李树静
ZHAO Zengyuan;GUO Weiting;BI Jianghua;FENG Chuntao;ZHU Hongbo;ZHANG Pu;SUN Jianhui;WANG Minsheng;YU Wenli;LI Shujing(Hebei Tianhe Beef Cattle Breeding Co.Ltd.,Shijiazhuang 050200,China;Hebei Cattle Industry Technology Research Institute,Shijiazhuang 050200,China;Shijiazhuang Animal Husbandry Technology Extension Station,Shijiazhuang 050200,China;Animal Health Supervision Institute of Luquan District of Shijiazhuang,Shijiazhuang 050200,China)
出处
《黑龙江动物繁殖》
2020年第5期8-13,共6页
Heilongjiang journal of animal reproduction
基金
石家庄市科学技术研究与发展计划项目(191500212A)
国家现代农业(肉牛牦牛)产业技术体系项目(CARS-37)
河北省技术创新引导计划项目(20826604D)
关键词
牛
胚胎分割
半胚发育率
半胚移植
妊娠率
双犊率
cows
embryo splitting
demi-embryo development rate
demi-embryo transfer
pregnancy rate
double calves rate
作者简介
赵增元(1983—),男,硕士研究生,研究方向为胚胎生物技术研发和推广,zhaozengyuanjsdx@163.com;通信作者:余文莉(1964—),女,研究员,博士,研究方向为胚胎生物技术研发和推广,anboyuwenli@126.com;通信作者:李树静(1964—),男,研究员,博士,研究方向为胚胎生物技术研发和推广,embryochina@163.com.
