胚胎因素对肉羊胚胎移植效果的影响

为提高肉羊胚胎移植的妊娠率,研究了胚胎发育阶段、胚胎体外保存时间和移植胚胎数对肉羊胚胎妊娠率的影响。结果表明:发育阶段对妊娠率有显著影响,在移植胚胎数量相同的情况下,囊胚的受胎率(58.7%)高于桑椹胚(56.4%)(P<0.05);胚胎体外保存0～10 min、10～30 min、30～60 min、60 min以上的妊娠率分别为66.7%、65.0%、60.0%、63.3%,四者间均没有显著差异(P>0.05);移植胚胎的数量对受胎率有显著影响,移植2枚囊胚的受胎率(67.7%)显著高于移植2枚桑椹胚(61.2%)、1枚囊胚(58.7%)、1枚桑椹胚的受胎率(56.4%)(P<0.05)。     

提高波尔山羊胚胎移植妊娠率研究

本文就波尔山羊胚胎质量、胚胎发育阶段、胚胎数量、受体卵巢黄体数、受体注射LRH-A3、供、受体发情同期化程度以及胚胎移植的方向等因素与受体移植妊娠率的关系进行了研究。结果表明:移植囊胚较移植1枚B级桑椹胚妊娠率高;A级桑椹胚妊娠率显著高于移植1枚B级桑椹胚(P<0.05);移植2枚B级桑椹胚妊娠率比移植1枚B级桑椹胚的高39.4%(P<0.05);受体注射LRH-A3有助于提高妊娠率;供、受体发情同步差在48 h以上组妊娠率显著低于0~24 h和24~48 h组(P<0.05);向子宫角前端方向的移植妊娠率比向子宫角基部方向的高12.2个百分点。     

影响和牛双半胚移植妊娠率因素分析

研究和牛胚胎不同胚龄和移植方式对分割双半胚移植妊娠率的影响。结果表明:致密桑椹胚、早期囊胚、囊胚分割后双半胚移植妊娠率分别为60.2%、53.8%和50.0%,随着胚龄的上升妊娠率有下降趋势,各组间无显著差异(P0.05);移植到受体牛同侧子宫角双半胚妊娠率高于子宫角双侧各移植半枚分割胚和整胚(58.6%、41.7%、51.2%)(P0.05),双侧移植双犊率显著高于单侧移植和整胚移植(85.7%、36.9%、0%)(P0.01)。     

性控精液对奶牛体内胚胎质量、发育和移植妊娠率的影响

[目的] 研究性控精液对奶牛体内胚胎质量、胚胎发育和胚胎移植妊娠率的影响。[方法] 将144头青年奶牛随机分为对照组（63头）和试验组（81头）,使用促卵泡激素（FSH,260 mg/头）进行超排处理。对照组和试验组分别使用常规精液和性控精液输精,并对获得的体内性控胚胎进行移植,对胚胎生产、胚胎质量、胚胎发育和胚胎移植妊娠情况进行统计。[结果] 试验组供体获得的平均可用胚胎数（5.67枚）显著（P<0.05）低于对照组（6.92枚）;试验组供体获得的可用胚胎中A级胚胎比例（62.53%）、B级胚胎比例（35.29%）与对照组（A级胚胎比例66.51%、B级胚胎比例30.97%）相比差异均不显著（P>0.05）;试验组供体获得的可用胚胎中桑葚胚比例（84.10%）显著（P<0.05）高于对照组（61.24%）,囊胚比例（15.90%）显著（P<0.05）低于对照组（38.76%）;试验组的鲜胚移植妊娠率（52.41%）显著（P<0.05）低于对照组（66.13%）。[结论] 与常规精液相比,使用性控精液生产奶牛体内性控胚胎并移植后,平均可用胚胎数、可用胚胎中囊胚比例和胚胎移植妊娠率降低,可用胚胎质量未明显降低;优化性控精液使用方案和胚胎移植技术能够提高体内性控胚胎生产和胚胎移植效率。     

影响荷斯坦奶牛胚胎移植成功率的因素

本文比较了不同术者和不同发育阶段的荷斯坦奶牛胚胎的移植妊娠率。在4个不同术者中,总胚胎移植妊娠率最高为67.1%,最低为37.5%,两者间差异极显著(P0.01);桑葚胚和早期囊胚的最高妊娠率为65.7%,最低为36.6%,两者间差异极显著(P0.01);囊胚的最高妊娠率为66.7%,最低为37.5%,两者间差异极显著(P0.01)。扩张囊胚的最高妊娠率为86.4%,最低为40.0%,两者间差异极显著(P0.01)。结论:不同术者在移植胚胎时,移植妊娠率存在着显著差异。4个术者合计的桑葚胚(50.2%)与囊胚(47.2%)的移植妊娠率差异不显著,但扩张囊胚的妊娠率相对较高(66.7%)。     

进口优质肉羊胚胎的移植试验

对从澳大利亚进口的304枚德克赛尔肉羊冷冻胚胎进行移植,产羔178只,产羔率为5855%.并对冷冻胚胎解冻、移植过程中的主要环节进行研究,其结果如下①解冻后的A、B级孵化囊胚所占比例为(43.75%)、桑椹胚--扩张囊胚解冻后的A、B级胚胎所占比例为(84.02%),两者有显著差异;②解冻后的A、B级桑椹胚--扩张囊胚移植产羔率(64.88%)显著高于解冻后的A、B级孵化囊胚移植产羔率(42.85%);解冻后的C级胚胎移植后也能得到一定的产羔率;③经产母羊胚胎移植产羔率(68.62%),显著高于育成母羊产羔率(51.85%).     

黑萨福克和杜泊羊新鲜和冷冻胚胎移植效率的研究

为提高优质肉用种用绵羊胚胎移植的效率,试验选取76只黑萨福克羊和27只杜泊羊作为供体,采用"CIDR+FSH+PMSG"法进行超数排卵处理。选取384只小尾寒羊为受体,采用"CIDR+PMSG"法进行同期发情处理,以探索供体胚胎发育阶段、胚胎冷冻处理、胚胎体外停留时间以及移植给受体时移植侧黄体数量等因素对胚胎移植妊娠率的影响。结果表明:移植新鲜囊胚的妊娠率极显著高于桑椹胚(P0.01);冷冻/解冻囊胚移植妊娠率极显著低于新鲜囊胚(P0.01),而冷冻/解冻桑椹胚移植妊娠率与新鲜桑椹胚无显著差异(P0.05);此外,移植桑椹胚时,移植侧有1个黄体的受体妊娠率极显著低于有2~3个黄体受体妊娠率(P0.01),但移植囊胚时两者间差异不显著(P0.05)。综上可知,子宫角移植桑椹胚和囊胚时,移植新鲜囊胚、移植冷冻桑椹胚效率较高;将新鲜桑椹胚移植给移植侧有2~3个黄体受体效果好于1个黄体受体。     

杜泊羊冷冻胚胎移植效果的影响因素及羔羊早期生长性能研究

为研究杜泊羊冷冻胚胎移植效果的影响因素以及羔羊早期的生长情况,本实验选用1 361枚质量合格的纯种黑头杜泊羊的早期冷冻胚胎作为供体,180只小尾寒羊和970只湖羊作为受体羊,采用同期发情和子宫角胚胎移植技术,研究季节(春季、秋季)、受体羊品种(小尾寒羊、湖羊)、卵巢上黄体个数(1个、2～3个)、受体羊的营养状况(中上等膘情、下等膘情)、胚胎发育阶段(桑椹胚、晚期桑椹胚、早期囊胚、囊胚和扩张囊胚)及移植胚胎数目(双胚和单胚)对于杜泊羊冷冻胚胎移植效果的影响。结果显示:秋季(11月)同期发情率显著高于春季(4月),但妊娠率没有显著差异;在春季,不同品种受体羊的同期发情率和妊娠率无显著性差异;受体羊卵巢上黄体个数对妊娠率无显著影响;中上等膘情受体羊的妊娠率显著高于下等膘情受体羊;移植扩张囊胚的受体羊妊娠率为30.0%,显著低于其他4个胚胎发育阶段;移植双胚的受体羊妊娠率低于移植单胚的受体羊。羔羊早期生长发育指标统计结果表明:在潍坊地区,小尾寒羊和湖羊均可以作为胚胎移植的受体,但是小尾寒羊体型偏大,每日饲料消耗偏多,湖羊作为受体羊在节约饲料方面更有优势。     

影响受体牛冷冻移植胚胎效果的主要因素

试验研究影响受体牛移植冷冻胚胎效果的主要因素,目的是解决胚胎移植产业化中容易出现的技术问题,以提高牛胚胎移植妊娠率。在季节、饲养管理、同期发情处理和胚胎移植技术以及环境条件等基本相同的情况下,结果表明:西门塔尔杂交牛的移植利用率和移植妊娠率高于黄牛杂交牛和黑白花奶牛,但各组间无显著性差异(P>0.05);2～6周岁的受体牛移植利用率和移植妊娠率高于7周岁以上和2周岁以下的受体牛,但各组间无显著性差异(P>0.05);经产1～3胎受体牛移植利用率显著高于4胎以上的受体牛(P<0.05),而移植妊娠率则无显著性差异(P>0.05);体况上等与中等的受体牛移植利用率和移植妊娠率无显著性差异(P>0.05);同期发情处理时卵巢处于黄体期的移植利用率显著高于卵泡期,差异极显著(P<0.01);左侧卵巢有黄体的子宫角胚胎移植妊娠率显著高于右侧(P<0.05);胚龄对胚胎移植妊娠率高低依次为早期囊胚、桑椹胚和囊胚,但无显著性差异(P>0.05)。     

进口优质肉羊胚胎的移植试验

对从澳大利亚进口的304枚德克赛尔肉羊冷冻胚胎进行移植，产羔178只，产羔率为58.55%，并对冷冻胚胎解冻，移植过程中的主要环节进行研究。其结果如下：（1）解冻后的A、B级孵化囊胚所占比例为（43.75%)，桑椹胚-扩张囊胚解冻后的A、B级胚胎所占比例为（84.02%)，两者有显著差异；（2）解冻后的A、B级桑椹胚-扩张囊胚移植产羔率（64.88%)显著高于解冻后的A、B级孵化囊胚移植产羔率（42.85%)；解冻后的C级胚胎移植后也能得到一定的产羔率；（3）经产母羊胚胎移植产羔率（68.62%)，显著高于育成母羊产羔率（51.85%)。     

Some Factors Affecting the Rate of Pregnancy after Embryo Transfer Derived from the Brazilian Jumper Horse Breed

This study aimed to evaluate the effects of certain embryo transfer parameters on the pregnancy rate after equine embryo transfer of the Brazilian Jumper Horse breed. The size, embryonic development stage, embryo quality, and synchronization of ovulation between the donor (n = 120) and recipient (n = 420) were evaluated in 396 embryos. Embryo recovery was performed on Day 6-9 after ovulation (Day 0 = day of ovulation). The recipient mares were chosen on the day of embryo recovery, and the transfers were performed that same day. The embryo size (diameter including envelopes; n = 396) ranged from 150 to 3000 μm; 67.1% measured between 400 and 1199 μm. The embryo size (400-1199 μm vs. ≤399 μm); stage of development (n = 396; blastocyst and expanded blastocyst versus compact morula and early blastocyst); quality (n = 396; grade 1 [excellent]), 2 [good], or 3 [poor]); and synchronization of ovulation between the donor and recipient (0, 1, 2, 3, and 4 days versus −1, 5, and 6 days, respectively) all affected pregnancy rate (P < .05). The pregnancy rate did not differ significantly among transfers performed on Days 0, 1, 2, 3, and 4. In conclusion, embryos measuring 400-1199 μm produced higher pregnancy rates in recipients than embryos measuring 150-399 μm, and blastocysts and expanded blastocysts produced pregnancy more efficiently than morulae and early blastocysts. The embryo quality also affected the pregnancy rate. Synchronization of donor and recipient ovulation to Days 0-4 improved the efficiency of embryo transplant.     

Japanese Society for Animal Reproduction: award for outstanding research 2002. The development and prevalence of the transfer technique for frozen-thawed embryos of Japanese black beef cattle in Tochigi Prefecture

The conditions of embryo transfer by the stepwise method, in which frozen-thawed embryos are transferred on day 7 (day 0=onset of estrus), were investigated with the aim of increasing pregnancy rates in frozen-thawed embryo transfer. The use of a vaginal speculum to prevent bacterial infection when passing an embryo transfer gun through the vagina yielded a pregnancy rate equal to or higher than that with application of a sheath cover to the transfer gun. Administration of a sedative, xylazine, to recipient cattle for preventing movement at the time of embryo transfer improved the pregnancy rate. The influence of the time from thawing of frozen embryos to transfer and of the transportation of the recipient by truck upon pregnancy rate was investigated. Embryo transfer within 60 minutes after aspiration into a straw or transportation of the bovine recipient, 1.5 hours each way before and after transfer, had no influence on pregnancy rate. Relations of the embryonic developmental stage and morphological quality after thawing of frozen embryos to pregnancy rate were investigated in recipients of nulliparous Holstein heifers. The pregnancy rate increased as the embryonic developmental stage advanced from compacted morula, early blastocyst, and blastocyst in that order. The pregnancy rate obtained with blastocyst stage embryos was significantly (P<0.05) higher than that with compacted morula stage embryos, and there was no significant difference in pregnancy rates between excellent morphological quality and good morphological quality for compacted morula stage embryos. When correlation of luteal function and pregnancy rate was investigated in bovine recipients, pregnancy rate showed a tendency to increase with increasing blood progesterone (P) concentration on the day before (on day 6 after estrus) and the day of embryo transfer. The pregnancy rate in bovine recipients, which showed a blood P concentration of > or =2.5 ng/ml on the day before embryo transfer, was significantly (P<0.05) higher than that in those with a blood P concentration of <2.5 ng/ml. Pregnancy rate showed a tendency to increase with decreasing blood estradiol-17beta (E2) concentration on the day of embryo transfer. Activation of luteal function by administration of human chorionic gonadotropin (hCG) in cycling cattle was investigated for its effect on increasing pregnancy rate in bovine recipients. A follicle coexisting with cyclic CL ovulated and induced CL formed after injection of hCG 1,500 IU 5 days after ovulation. The blood P concentration was significantly (P<0.05) higher in the administration group than in the control group, and the blood E2 concentration rapidly decreased, showing a lower concentration than in the control group. These results suggest the possibility that the pregnancy rate could be improved by administration of hCG. Pregnancy rate following intramuscular injection hCG 1,500 IU was comparatively investigated in parous Japanese Black beef cattle receiving frozen-thawed embryos 7 days after estrus. Pregnancy rate was 67.5% in the group in which hCG was administered on day 6 after estrus, and was significantly (P<0.05) higher than that in the control group (45.0%) and the group in which hCG was administered on day 1 after estrus (42.5%), revealing that hCG administration facilitated pregnancy. Transfer of frozen-thawed embryos in the blastocyst stage within 60 minutes after the aspiration into a straw, with a vaginal speculum after administration of xylazine is suggested as a way of improving pregnancy rate in bovine recipients with favorable luteal function and in those with luteal function activated by administration of hCG on the day before embryo transfer.     

Transfer of Bisected Cattle Embryos within the Same Zona Pellucida*

Cattle embryos at the compact morula and blastocyst stages were bisected by two different methods. In some embryos the embryo bisection was performed outside of the zona pellucida and both halves were replaced into it. Other embryos were bisected within the zona pellucida and no pipetting of the halves was required. They were transferred to synchronized recipient cows and the pregnancy rates and twinning rates were evaluated.     

不同品种绵羔羊超数排卵及胚胎体外生产技术研究

探讨品种对羔羊超数排卵效果和卵母细胞发育能力的影响,并将体外受精胚胎进行了移植,为研究利用绵羊羔羊卵母细胞生产后代提供理论和技术方法。对4～8周龄不同品种羔羊用FSH+PSMG处理后获得的平均卵母细胞数量和体外胚胎卵裂率和囊胚率进行了比较,并比较成熟液中有无β-巯基乙醇对胚胎卵裂率和囊胚率的影响。结果:各品种获得的平均卵母细胞数分别为:哈萨克为210.22枚,湖羊为0枚,美利奴为135.80枚,湖羊与哈萨克的杂交后代为37.80枚,湖羊羔羊和湖羊与哈萨克羊的杂交羔羊获得的平均卵母细胞数显著低于哈萨克羔羊(P<0.01,P<0.05)。哈萨克羔羊卵裂率(69.0%)显著高于湖羊与哈萨克羊杂交羔羊(66.4%,P<0.05);哈萨克羔羊囊胚率(17.5%)和美利奴羔羊囊胚率(16.2%)显著高于湖羊×哈萨克羔羊(13.8%)(P<0.01,P<0.05)。β-巯基乙醇对羔羊胚胎卵裂率影响不显著,对囊胚率影响极显著(P<0.01)。哈萨克羔羊卵子体外受精获得的2-8细胞胚胎进行输卵管移植,14只受体羊中7只妊娠,妊娠率为50.0%;共产羔9只,其中8只存活。     

影响绵羊冻胚移植妊娠率的因素分析

本研究对胚胎质量、胚胎移植数量、胚胎发育阶段、冷冻方法、移植方法及饲养管理水平等因素对移植产羔率的影响进行研究,以提高绵羊胚胎移植效率。结果表明:采用1.5mol/L乙二醇做冷冻保护剂对胚胎进行常规冷冻,冻前A级胚胎冷冻/解冻后可用胚率、存活率、降级率均高于B级,差异极显著(P<0.01)。移植1枚冷冻解冻后A级、B级或2枚(A+B)胚胎的受体移植产羔率分别为44.48%、46.84%和44.81%。经χ2检验分析,三者之间无显著差异(P>0.05)。移植双胚的受体双羔率为22(%53/241),以胚胎为单位计算,产羔率仅为33.40%。A级囊胚和桑椹胚的产羔率之间无显著差异(P>0.05)。1.5 mol/L乙二醇常规冷冻保存的体内胚胎移植产羔率高于EFS40玻璃化冷冻保存,但经统计学分析,二者无显著差异(P>0.05)。子宫手术移植和腹腔内窥镜法子宫移植之间产羔率不具统计学差异(P>0.05)。在牧场条件下大规模移植的平均产羔率为43.95%,范围在20%￣70%之间,受体饲养管理水平对绵羊移植产羔率有较大影响。     

细胞松弛素B对猪孤雌胚胎和体外克隆胚胎发育能力的影响

为探讨细胞松弛素B（cytochalasin B,CB）对猪孤雌胚胎和克隆胚胎发育能力的影响,本研究通过在猪体外胚胎培养基中添加不同浓度CB以及不同孵育时间的处理,筛选出CB对猪早期胚胎发育的最适浓度和最佳孵育时间,同时通过Hoechst33342染色检测猪体外囊胚孵化期的细胞数差异,进一步研究CB对孤雌胚胎和克隆胚胎发育的影响。结果显示,培养基中添加CB浓度为7.5 μg/mL时孤雌胚胎和克隆胚胎的卵裂率分别为85.00%和90.23%,囊胚率为35.68%和42.58%,均显著高于其他各组（P < 0.05）;采用7.5 μg/mL CB处理电激活后的孤雌胚胎和克隆胚胎,孤雌胚胎孵育4 h组的卵裂率（83.80%）和囊胚率最高（35.39%）,与其他各组差异显著（P < 0.05）,而克隆胚胎孵育6 h组的卵裂率（83.98%）和囊胚率最高（55.62%）,与其他各组差异显著（P < 0.05）。此外,Hoechst33342染色结果显示,未添加CB处理的孤雌胚胎在囊胚孵化期的细胞平均数为28个,CB处理组的孤雌胚胎和克隆胚胎细胞平均数分别为36和52个,处理组和未处理组细胞数差异显著（P < 0.05）。结果表明,猪体外孤雌胚胎用7.5 μg/mL CB 处理4 h可获得较高的卵裂率和囊胚率;体外克隆胚胎用7.5 μg/mL CB 处理6 h卵裂率及囊胚率最高,且囊胚期内细胞团细胞总数最多。CB处理有利于体外胚胎早期发育,提高克隆胚胎移植受孕率。     

奶牛活体采卵-体外受精效率的影响因素研究

为建立稳定高效的活体采卵-体外受精技术体系,提高体外胚胎生产效率,本研究先利用屠宰场采集的新鲜卵巢卵母细胞进行体外受精,通过胚胎发育潜力来筛选最佳的体外胚胎培养液;再进一步研究不同种公牛精液和供卵母牛对活体采卵-体外受精效率的影响。结果显示,CR1aa培养液和mCR1aa培养液卵裂率差异不显著（P>0.05）,但mCR1aa组的囊胚发育率显著提高（28.1% vs 20.6%,P<0.05）;选取的3头荷斯坦种公牛精液的活体采卵-体外受精胚胎的卵裂率差异不显著（P>0.05）,但1号种公牛精液体外受精后囊胚率（38.7%）显著高于2号和3号（23.8%&22.9%）（P<0.05）;随机选择的3头活体采卵供体母牛（H1、H2、H3）获得的头均可用卵母细胞数无显著差异,但H1和H2供体母牛体外受精胚胎的卵裂率和囊胚率均显著高于H3供体牛（P<0.05）,且H1供体牛体外受精囊胚率显著高于H2供体牛（P<0.05）。结果表明,mCR1aa培养液能显著提高体外受精囊胚发育率,适用于体外胚胎生产;种公牛精液和供体母牛个体差异会直接影响活体采卵-体外受精胚胎的生产效率,为奶牛活体采卵-体外受精生产技术体系的优化提供参考。     

Proteomic analysis demonstrates that parthenogenetically activated swamp buffalo embryos have dysregulated energy metabolism

The comprehensive understanding of early embryo development is essential to optimize in vitro culture conditions. Protein expression landscape of parthenogenetically produced embryo remains unexplored. This study aimed to investigate the protein expression dynamics with a particular focus on energy metabolism throughout the early developmental stages of parthenogenetic buffalo embryos. For this purpose, we performed iTRAQ-based quantitative mass spectrometry and identified 280 proteins common in all stages. A total of 933 proteins were identified during the proteomics analysis. The data depicted that morula and blastocyst had distinct protein expression dynamics as compared to 2- to 16-cell-stage embryo. KEGG pathway analysis showed 23 proteins belonging to energy metabolism appeared in the data. Study of energy metabolism-related protein's expression pattern demonstrated that there was asynchrony in proteins related to glycolysis throughout the examined developmental stages. The expression pattern of pyruvate kinase mutase (PKM), an essential protein of glycolysis, indicated a slightly decreasing trend from 2-cell-stage embryo to blastocyst, and it was supported by expression of proteins involved in lactate production (LDHA and LDHB) suggesting the decreasing rate of aerobic glycolysis (Warburg Effect) at morula and blastocyst stage. The increased Warburg Effect is considered as the hallmark of proliferating cells or embryo at the blastocyst stage. Furthermore, the proteins involved in the citric acid cycle also showed down-regulation at the blastocyst stage, indicating a lesser role of oxidative phosphorylation at this stage. Therefore, it could be divulged from the study that there may be an irregular pattern of energy metabolism in early parthenogenetic embryos. Further studies are recommended to understand this phenomenon.     

黄牛卵母细胞和早期胚胎组蛋白乙酰化转移酶1表达模式研究

为探讨卵母细胞成熟及早期胚胎发育过程中组蛋白乙酰基转移酶(HAT1)的表达规律,研究应用实时定量PCR技术,检测了广西本地黄牛卵母细胞和附植前胚胎HAT1基因的表达情况。结果表明:HAT1基因在黄牛生发泡期(GV)卵母细胞、第2次减数分裂中期(MⅡ)卵母细胞、体外受精(IVF)胚胎2～4细胞、8～16细胞、桑葚胚和囊胚中的相对表达量分别为1.00、0.56、0.08、0.55、0.43和0.31,在孤雌激活(PA)胚胎的2～4细胞、8～16细胞、桑葚胚和囊胚中的相对表达量分别为0.55、0.55、0.48和0.46。HAT1在GV期相对表达量最高,在IVF胚胎中2～4细胞表达量最少(P<0.05)。由此可见,HAT1基因在黄牛卵母细胞成熟和早期胚胎阶段均有表达,GV期HAT1基因的表达最高,PA胚胎HAT1基因的表达较稳定。