摘要
本研究着重探讨和研究了奶牛活体采卵(ovum pick-up,OPU)及体外受精(in vitro fertilization,IVF)各技术环节中一些因素对奶牛OPU和IVF整个过程的影响,初步建立了一套比较完善的操作技术体系。研究结果如下:
OPU试验:实验一,不同采卵间隔时间对奶牛OPU的影响:分别对奶牛进行活体采卵2次/周和1次/周。结果如下:每周2次平均获卵数要多于每周1次,但差异不显著(5.14、4.66;P>0.05);优质卵母细胞率2次/周和1次/周比较,差异极显著(80.5%、65.9%;P<0.05)。实验二,激素处理与非激素处理组对OPU的影响:处理组的平均获卵数显著高于对照组(8.56、4.50; P<0.05)差异显著,优质卵母细胞率(72.6%、79.2%;P>0.05)差异不显著。实验三,青年牛(14～16月龄)与成年牛对OPU的影响:青年牛和成年牛的平均获卵数(5.89、4.41; P>0.05)和优质卵母细胞率(78.2%、72.3%;P>0.05)高于成年牛,但差异不显著。实验四,健康牛与子宫疾病牛对OPU的影响:健康牛平均获卵数(5.92、3.88;P<0.05)和优质卵母细胞率(81.1%、69.9%;P<0.05)均显著高于和患有子宫疾病的牛只。实验五,高频率连续活体采卵对采卵效果的影响:为了研究高频率连续活体采卵对采卵效果的影响,分别对奶牛连续采卵和只采集一次。实验结果显示:对奶牛连续活体采卵和只采集一次的平均获卵率(5.16、5.80;P>0.05)和优质卵母细胞率(80.6%、84.5%;P>0.05)差异均不显著。
IVF实验:实验一,不同来源的卵母细胞对奶牛IVF的影响:分别对采用屠宰场采集的卵母细胞和活体采卵收集的卵母细胞进行体外培养,受精后36h和7d后观察卵裂率和囊胚发育率,两组的的卵裂率(77.6%、81.3%;P>0.05)和囊胚发育率(25.5%、29.2%;P>0.05)差异不显著。说明卵母细胞来源对IVF的影响不显著。实验二:对精子分别采用percoll梯度密度离心和上浮法处理,两组的卵裂率(80.7%、73.7%;P>0.05)没有明显差异,囊胚发育率(27.6%、19.7%;P>0.05)虽有所提高,但差异不显著。实验三,不同培养体系对奶牛IVF的影响:分别用SOF液和BO液作为体外培养系统。从表中可以看出,两者培养体系的卵裂率(82.0%、77.9%;P>0.05)和囊胚发育率(31.1%、26.7%;P>0.05)均差异不显著。实验四,受精时间分别采用8h、10h、12h。三组的卵裂率(83.6%、79.2%、75.7%;P>0.05)逐渐降低,但差异不显著。囊胚发育率受精时间8h、10h(30.3%、26.4%;P>0.05)及10h、12h(26.4%、19.6%;P>0.05)均差异不显著,但受精8h的囊胚发育率显著高于受精时间12h(30.3%、19.6%;P<0.05)。实验五,受精微滴(100μl)中放置成熟的卵母细胞数分别为20、40个,两组的卵裂率(82.0%、70.5%;P<0.05)和囊胚发育率(31.0%、21.5%;P<0.05)均下降,并且差异显著。
The aim of this study was to establish an steady and efficient(ovum pick-up,OPU)and(in vitro fertilization, IVF)system in our lab.Experiments were carried out to analyze the different factors reported influence the results of OPU and IVF in dairy cattle.
Study of OPU:1.The compare of oocyte taking from dairy cattle twice and once per week.The average number of oocytes (5.14、4.66;P>0.05)was not significantly different between the two groups;whereas,the high quality oocyte rates(80.5%、65.9%;P<0.05)twice per week were higher than once per week.2.The dairy cattle treated by hormones and untreated.The average number of oocytes(8.56、4.50; P<0.05) treated were higher than the untreated;whereas,The high quality oocyte rates(72.6%、79.2%;P>0.05)were not significantly different between the two groups.3.The differences of oocytes taken by OPU in heifers or dairy cattle.The average number of oocytes(5.89、4.41;P>0.05)and high quality oocyte rates(78.2%、72.3%;P>0.05)were not significantly different between the two groups.4.The differences of oocytes taken by OPU in healthy and not healthy dairy cattle.There were significantly different in recovered oocytes(5.92、3.88;P<0.05)and high quality oocyte rates(81.1%、69.9%;P<0.05)between the two groups.5.Dairy cattle oocytes collected repeatedly and once.There were no significant differences in recovered oocytes(5.16、5.80;P>0.05)and at high quality oocyte rates(80.6%、84.5%;P>0.05)between the two groups.
Study of IVF:1.IVF with oocytes from OPU and slaughterhouse.The results showed that cleavage rates(77.6%、81.3%;P>0.05)and blastocyst rates(25.5%、29.2%;P>0.05)were not significantly different between the two groups.2.The sperm were treated by percoll and upriver.There were no significant differences at cleavage rates(80.7%、73.7%;P>0.05)and blastocys rates(27.6%、19.7%;P>0.05)between the two groups.3.The effects of different culture systems(SOF、BO)on IVF of dairy cattle.The cleavage rates(82.0%、77.9%;P>0.05)and blastocys rates(31.1%、26.7%;P>0.05)in SOF were not significantly different between the two groups.4.Cleavage rates were(83.6%、79.2%、75.7%;P>0.05)respectively at the fertilization time 8h、10h and 12h.There were no significant differences between the three groups.The blastocyst rates at 8h were higher than at12h(30.3%、19.6%;P<0.05)。But the blastocyst rates were not significantly different between 8h and 10h(30.3%、26.4%;P>0.05)and between 10h and 12h(26.4%、19.6%;P>0.05).5.The different oocytes number added to fertilization dripping(20、40).The results showed that the former cleavage rates(82.0%、70.5%;P<0.05)and blastocyst rates(31.0%、21.5%;P<0.05)was higher than the latter.
引文
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