奶牛超数排卵方法初探

超数排卵效果的好坏直接关系到胚胎移植技术在畜牧业中的应用。为此广大畜牧工作者对超排药物的种类及其之间的比较、处理方法以及重复处理 的间隔时间等都进行了广泛深入的研究,使母牛的排卵数由过去的平均2～6枚到今天的6～10枚或更多。本文仅对奶牛在性周期的不同阶段对其实施超数排卵 的诸种方法加以介绍。     

奶牛超数排卵的研究（第二报）

奶牛超数排卵是胚胎移植最重要的基础环节。只有稳定的超排方法才能提供充足的卵源,从而加速胚胎移植技术在生产中的应用。长期以来,许多人对牛的超排方法和影响超排效果的若干因素进行过大量研究,但超排效果仍不稳定,个体间存在明显的差异性。就一局部地区而言,寻找一种适合于当地条件、超排效果相对稳定的处理程序和方法,已是人们极为关注的研究课题。本试验目的是研究不同激素制剂处理方法,不同年龄和重复处理等因素对奶牛超排效果的影响,在1988年采卵供体平均获可用胚胎5.0枚的基础上,1989年采用较合理的超排处理方案,使每头采卵供体得可用胚胎提高到6.10枚,为探讨效果稳定的超排方法提供了依据。     

超数排卵和胚胎移植技术在奶牛繁育中的应用

超数排卵和胚胎移植技术是一项日益成熟的生物工程技术，本实验采用MOET技术，以黄牛为受体，推广高产奶牛，取得了较好的效果。     

奶牛胚胎远距离运输移植黄牛技术研究

超数排卵处理黑白花奶牛１１头,有效８头,超排有效率７２．７％,获总卵数４０枚,头均获卵５枚（４／８）,可用胚２６枚,头均可用胚３．３枚（２６／８）,胚胎可用率６５％。移植受体牛２３头,妊娠１０头,移植成功率４３．５％（２３／１０）。     

核心群奶牛超数排卵技术影响因素的研究

[目的]为进一步研究核心群奶牛超数排卵技术影响因素。[方法]试验使用3个FSH剂量和4 个公牛精液对27头青年奶牛进行超数排卵。对供体牛胚胎生产情况和受体胚胎移植妊娠率进行统计。[结果]结果显示供体2次超排获得7.8枚平均可用胚胎显著高于1次超排获得的5.7枚平均可用胚胎(P<0.05);260,280 mg FSH超排供体获得6.0和7.7 枚平均可用胚胎显著高于300 mg FSH超排供体获得的4.6枚平均可用胚胎(P<0.05);供体使用4个公牛精液人工授精后获得的平均可用胚胎数和平均未受精卵数差异不显著(P>0.05);移植前接种疫苗的受体胚胎移植妊娠率显著降低(P<0.05）。[结论]核心群奶牛超排适宜FSH剂量为260-280 mg,使用高受精力公牛精液和对超排反应好的供体重复超排能够提高超排效率,受体移植前避免疫苗接种能够提高胚胎移植妊娠率。     

不做发情检测对奶牛进行超数排卵的新方法

超数排卵(超排)的目的是获得最大数量的可移植胚胎,在传统的超排方案中,处理于发情周期的中期(排卵后8￣12天,等于发情后9￣13天),奶牛在超排前,需要通过直肠检查卵巢黄体的质量好坏才能确定是否选用。其一是增加了劳动强度;其二是一部分牛因排卵后没能形成好的黄体而未被利用,而且超排处理一般在产后60天进行,重复超排间隔时间延长到2个月、甚至半年以上,严重影响了高产奶牛的利用率,减少了可移植胚胎的数量。目前,我们采用通过控制卵泡波同期化方法,在奶牛发情后的任意1天,在不做发情检测的情况下,在供体牛阴道内放置CIDR(孕酮阴道硅胶…     

奶牛超数排卵技术研究进展

奶牛超数排卵是胚胎移植技术的重要环节之一。牛超数排卵的目的是最大限度地获得受精卵和可用胚胎（Foote et al．，1986），然而，在实际生产中牛的超排反应和胚胎质量差异性很大。卵巢反应的差异性受许多因素的影响，如促性腺激素种类（Elsden et al．，19781、批号（Murphy et al．，1984）、处理时间（Lindsellet al．，1986）、剂量（Pawlyshyn et al．，1986）、季节（Massey et al．，1984）、牛的品种（Saumande et al．，1978）、年龄（Hasler et al．，1981）、处理时的卵巢生理状态（Moor et al．，1984）、营养状况（Hendricks et al．，1986）、繁殖史Hasler et al．,1983）和重复超排次数（Donaldson et al．，1983）等。超排就是在外源生殖激素作用下对母牛生殖系统所产生的一系列复杂生理过程。     

高产奶牛不同超数排卵方法的试验研究

本试验采用了不同方法对高产荷斯坦奶牛供体进行超数排卵。结果表明：（1）在自然发情周期的第3天、第6天、第9天和第12天分别开始用超排激素Folltropin～V处理，结果以第9天开始超排反应最佳；（2）应用含孕激素阴道栓＋雌激素＋孕激素处理后超排效果显著高于单独使用阴道拴组和阴道栓＋孕激素组，说明雌激素对超排具有明显促进作用；（3）比较应用进口阴道栓CIDR处理和国产“牛欢”配合雌激素和孕激素处理后进行超排，对青年牛和成母牛超排反应均没有差异，证明用国产阴道海绵栓可以代替进口的CIDR；（4）短期重复超排（间隔25～28天）处理次数对荷斯坦母牛的超排效果没有显著影响，但与传统方法比较，应用阴道拴（CIDR或牛欢）诱导卵泡波同步发生后开始超排处理，可以提高重复超排处理次数，提高超排效率，增加胚胎年产量。     

奶牛胚胎移植黄牛应用技术研究

超排处理黑白花奶牛58头,有效47头,头均获卵7.2±5.2枚(340/47),可移胚4.7±4.3枚(220/47)。两种超排方法比较试验,低剂量预处理超排组头均获卵8.2±5.3枚(164/20),可移胚5.2±4.3枚(103/20),高于常规超排组头均获卵数6.5±5.1枚(176/27),可移胚数4.3±4.3枚(117/27)。鲜胚移植农户饲养的黄牛受体,其受胎率为44.6％(41/92)。     

不同来源的促性腺激素对奶牛超数排卵的影响

目前，我国奶牛、肉牛胚胎移植工作已广泛开展，胚胎来源主要依靠进口，国内自给的超排取卵效果不稳定，不能尽快形成社会生产力。笔者通过利用不同来源促性腺激素FSH，对进口澳牛做了超数排卵效果试验。为该项技术的推广和应用提供了经验和科学依据。     

牛超排技术的研究

用４组药物组合：（１）ＰＭＳＧ＋ＰＧＦ２ａ；（２）ＰＭＳＧ＋ＰＧＦ２ａ＋激光照射；（３）ＦＳＨ＋ＰＧＦ２ａ；（４）ＦＳＨ＋Ｃｔｏｐｒｏｓｔｅｎｏｌ，超排处理３２２头次供体牛。共采卵１６６３枚（头均５．８１±４．６８），可用胚胎９８０枚（头均３．４３±３．９４，占５８．９％）。（３）、（４）两组胚胎可用率及可用胚胎数高于（１）、（２）组（ｐ＜０．０５）；黄牛胚胎可用率高于奶牛（ｐ＜０．０１）；育成牛胚胎可用率高于经产牛（ｐ＜０．０１）；供体牛前两次超排效果更好（ｐ＜０．０５）。     

荷斯坦母牛超数排卵及胚胎移植试验初报

应用FSH对3头荷斯坦母牛进行超数排卵处理，在母牛发情后第7天，2头共冲出11枚卵子，其中受精卵2枚（1枚桑椹胚，另1枚退化）。将回收的胚胎移植给1头受体牛，2个月后直检怀孕。     

青年荷斯坦奶牛初次超数排卵试验

对影响荷斯坦青年奶牛超数排卵的因素进行了对比分析,并探索了青年奶牛超排的可行性方案.卵巢上有黄体存在的供体牛可以用于超排.青年奶牛在不同性周期阶段进行超排的结果不同,但差异不显著.用FSH联合氯前列烯醇,采用常规逐日递减法肌肉注射进行超排,平均获得胚胎7.07枚,可用胚胎5.98枚;在人工授精的同时注射促排卵素3号(LRH A3),平均回收胚胎8.64枚,可用胚胎为5.5枚.促排卵素3号能提高胚胎的回收率,但可用胚胎率急剧下降.     

牛胚胎移植产业化超数排卵的系统研究

为了提高供体牛的利用率,加速胚胎移植技术的产业化,试验系统地研究了超排方案、青年奶牛的月龄及发情周期不同阶段、重复超排次数、超排间隔时间、卵巢状态、胎次与产后间隔时间对超排效果的影响,共超排供体奶牛1 127头次,回收胚胎共10 900枚,可用胚胎数量为6 942枚,平均每头次获得胚胎数量为6.73枚.结果表明:①应用PSO1、PSO2和 PSO3三种方案进行超排处理,PSO3方案的超排成功率和平均回收可用胚胎数均高于其他两个方案;②利用发情周期9～11 d的自然发情奶牛直接超排效果最好;③对青年奶牛进行超排处理,应限制超排起始时间,15月龄以上的青年奶牛超排效果较好;④供体牛连续重复超排控制在3次以内,连续4次超排处理后极大降低胚胎可用率;⑤超排处理的间隔最短时间应该选择46～60 d;⑥经产牛1～3胎次超排效果较好,7胎次以上较差;⑦产后间隔时间选取80～90 d.     

A new method for bovine embryo production: a potential alternative to superovulation

Eight cows were used to study the feasibility of transvaginal ultrasound-guided puncture of follicles as a method for the collection of immature oocytes for embryo production in vitro. In six trials at intervals of seven days, 104 oocytes were collected. After in vitro maturation and fertilisation the 104 oocytes were transferred to the oviducts of sheep. Six days later, 75 oocytes were recovered by flushing the oviducts. Twenty-four per cent of the recovered oocytes/embryos had developed into transferable and viable morulae and, or, blastocysts. The data show that this non-surgical and repeated collection of immature oocytes can be used successfully for the in vitro production of bovine embryos. The procedure may produce yields of embryos comparable to those obtainable by conventional superovulation procedures.     

广西西门塔尔青年母牛和荷斯坦母牛超数排卵及胚胎移植初报

本试验于 2 0 0 2年 1 2月 1 6日至 2 0 0 3年 2月 2 8日期间在广西畜牧研究所种牛场对 3头西门塔尔青年母牛和 5头荷斯坦母牛作了超排 ,同时做供体牛及受体牛的同期发情 ,杂种牛和本地牛的胚胎移植和胚胎冷冻试验 ,并获得较好的效果 ,现报告如下。1　材料与方法1 .1　供体牛与受体牛的选择供体牛 :选择本所种牛场引进的纯种西门塔尔青年母牛 3头 ,平均年龄 2 0个月 ,体重达3 50千克以上 ,已正常发情 3次以上 ,膘情中上等。荷斯坦母牛 5头 ,其中经产母牛 2头 ,年龄3 -5岁 ,产奶为 60 0 0千克以上 ,青年母牛 3头 ,平均年龄 2 0个月 ,体重达 3…     

Studies on factors affecting superovulation and embryo transfer in Hungarian merino ewes

The objectives of this study were (a) to assess the ovulatory response and embryo production of Hungarian Merino ewes after superovulation, (b) to investigate the factors influencing the efficiency of embryo transfer (ET) in Hungarian Merino ewes, (c) to compare the results of two ovarian stimulation protocols (PMSG and PMSG + FSH treatment) in Hungarian Merino ewes, and (d) to study how superovulation, laparoscopic insemination and surgical embryo retrieval (ER) affect the subsequent reproduction of Hungarian Merino donor females after an ET programme. There was no significant difference between the ovarian stimulation protocols in the ratio of donor ewes responding to superovulation nor in the average number of corpora lutea. However, the number of transferable embryos recovered per donor ewe was higher in the PMSG + FSH group. The proportion of transferable embryos, unfertilized oocytes and degenerated embryos did not differ between the treatment protocols. The total pregnancy rate was 53.4% (179/335). Neither the developmental stage of the embryo nor the number of transferred embryos affect the implantation of embryos. However, the increased number of transferred embryos positively influenced the pregnancy rate. No difference was found in the pregnancy rate between synchronised and non-synchronised groups of recipients. Thirty-six out of 45 donor ewes (80%) became pregnant within one year after the ET programme, indicating that ovarian stimulation and surgical ER did not affect adversely their reproduction.     

Association analysis between variants in bovine progesterone receptor gene and superovulation traits in Chinese Holstein cows

The objective of this study was to identify a predictor to forecast superovulation response on the basis of associations between superovulation performance and gene polymorphism. The PCR-RFLP method was applied to detect two reported single nucleotide polymorphisms (SNPs) of G59752C and T81637C (rs41614030) located in introns 3 and 4 of the bovine progesterone receptor (PGR) gene in 171 Chinese Holstein cows treated for superovulation and evaluate its associations with superovulation traits. In polymorphic locus 81637, all cows without superovulation response were g.81637TC and g.81637TT genotypes. Association analysis showed that these two SNPs had significant effects on the total number of ova (TNO) (p<0.05), and the T81637C polymorphism was significantly associated with the number of transferable embryos (p<0.05). In addition, significant additive effects (p<0.05) on TNO were detected in the polymorphisms of G59752C and T81637C. These results showed for the first time that the G59752C and T81637C polymorphisms in PGR gene were associated with superovulation traits and indicated that PGR gene can be used as a predictor for superovulation in Chinese Holstein cows.     

牛体外受精胚胎移植条件的研究

通过体外受精技术和胚胎移植技术充分利用现有优秀种公牛资源,大量生产优质胚胎,可以充分发挥良种奶牛的繁殖潜力,加快良种奶牛繁殖速度[1].因此开展体外胚胎生产的研究,探索出一套简单有效的体外胚胎生产技术,找到适合于体外胚胎的移植方法和时机,则无论对于目前养牛业发展还是对纯粹的科学研究都是非常必要的.本研究对奶牛胚胎移植做了全面的研究和探索,希望能形成一套简单规范的体外胚胎生产和移植的技术,以满足科研和生产的需求.     

Evaluation of the relationship between serum paraoxonase-1 activity and superovulation response/embryo yield in Holstein cows

In this study, the effect of serum paraoxonase-1 (PON-1) activity on superovulation response and embryo yield was evaluated. The study material comprised 50 Holstein cows aged 3–4 years on postpartum day 90–120 with a body condition score of 3–3.25. A progesterone-based estrus synchronization protocol was initially administered to the selected donors. For this purpose, progesterone source was inserted intravaginally (day 0) and gonadotropin-releasing hormone injection was performed (day 6). Seven days after the insertion of progesterone device, follicle-stimulating hormone injections (total dose of 500 µg in decreasing doses for 4 days) were administered for superovulation. On the morning of the ninth day, prostaglandin (PG) F2α was administered, and the progesterone device was removed from the vagina in the evening on the same day. Two days after PGF2α administration, fixed-time artificial insemination was performed in the morning and in the evening. On the day of artificial insemination, blood samples were taken from the donors to determine the serum PON-1 activity. Uterine flushing was performed seven days after insemination. The results revealed that the serum PON-1 activity (mean ± SD, 562.71 ± 140.23 U/l) of the cows that responded to superovulation (donors with total corpus luteum count of ≥3 in both ovaries) was higher than those (389.91 ± 80.51 U/l) that did not (P<0.05). On the day of insemination, a positive correlation was determined between serum PON-1 activity and the counts of total corpus luteum (r=0.398), total oocyte/embryo (r=0.468), transferable embryo (r=0.453), and Code I embryos (r=0.315, P<0.05). Unlike the Code I embryos, there was no significant correlation between serum PON-1 activity and the number of Code III embryos. Moreover, no significant difference in the number of Code III embryos between the two PON-1 groups was observed. However, embryo yield and quality were found to have increased with increased PON-1 activity. Therefore, it was concluded that serum PON-1 activity may be associated with superovulation response, embryo yield and quality in donor cows.