波尔山羊胚胎移植影响因素的研究

试验就胚胎的发育阶段、胚胎保存时间及移植胚胎数等因素对胚胎移植效果的影响情况进行了研究.结果表明,胚胎发育日龄对受体妊娠率没有显著影响(P＞0.05),但移植8～16细胞的胚胎的妊娠率略高于2～4细胞(58.49% vs 61.47%).人员不是影响受体妊娠率的主要因素.胚胎在体外保存0～120 min对妊娠率没有显著影响,但以在体外保存0～30 min内移植的妊娠率最高(65.61%).黄体与移植胚胎的相对位置对受体的妊娠率没有显著影响(P＞0.05),但移植双胚的受体的妊娠率较移植单胚高(61.76% vs 45.17%).同时,试验统计了经胚胎移植产生的羔羊和自然繁殖羔羊之间的差异,发现经胚胎移植产生的波尔山羊母羔的初生重极显著地高于自然繁殖的母羔(P＜0.01),但公羔间的初生重以及公、母羔间的体高、体长、胸围、死亡率和性别比例与自然繁殖羔羊差异不显著(P＞0.05).     

性别鉴定后胚胎移植妊娠率影响因素分析

对影响后的性别鉴定胚胎移植妊娠率的因素进行了分析。结果表明,性别鉴定胚胎的鲜胚和冻胚移植妊娠率均低于非性别鉴定胚胎,但二者差异不显著(P>0.05),这表明性别鉴定胚胎在生产中应用是可行的。随着胚胎细胞取样数的增多,其移植妊娠率逐渐下降,且冷冻带来的损伤明显增大;性别鉴定胚胎在体外放置时间小于5 h的妊娠率比大于5 h的高,且差异极显著(P<0.01);性别鉴定胚胎移植时受体的最佳移植时间在发情后的7.5~8.5 d;繁殖力好的受体牛移植妊娠率比繁殖力差的受体牛移植妊娠率高,且差异极显著(P<0.01)。     

解冻后胚胎质量对移植妊娠率的影响

研究分析了胚胎质量等级对胚移妊娠率的影响,用于移植的冷冻性控胚胎由加拿大进口,胚胎移植的受体为鲁西黄牛和西门塔尔牛。分析结果显示:解冻后的胚胎等级对受体牛的妊娠率有一定程度的影响,其中A级胚胎的受体妊娠率较C级胚胎的受体妊娠率高15%左右,差异显著(P<0.05);另外,虽然受体母牛品种对移植受胎率也有一定程度的影响,但没有显著差异(P>0.05)。     

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。     

胚胎因素对肉羊胚胎移植效果的影响

目的:观察分析胚胎因素对肉羊胚胎移植效果的影响.方法:选择258只已产1胎营养水平中上等的受体母肉羊为研究对象,对其移植A级胚胎,对比胚胎因素对肉羊胚胎移植效果的影响.结果:移植1枚桑椹胚的受胎率显著高于移植1枚囊胚者,不同体外保存时间的受体羊受胎率差异不具有统计学意义(P>0.05),随着胚胎移植数量的增长,受胎率明显提高(P<0.05).结论:胚胎因素与肉羊胚胎移植效果密切相关,其中胚胎发育阶段、移植胚胎数对受胎率有显著影响,胚胎体外保存时间对受胎率无显著影响,分析相应的影响因素对提高受胎率具有重要价值.     

移植波尔山羊胚胎对本地山羊某些生产指标的影响

通过对161只胚胎移植受体羊的分娩情况、羔羊初生重进行统计分析,结果表明,移植后本地山羊的平均妊娠期为150.9±1.8d,羔羊平均初生重为3.9±0.5kg,分别与本地山羊自然繁殖妊娠期和纯种波尔山羊自然繁殖羔羊初生重相比无明显差异(P>0.05)。但经胚胎移植的公羔羊初生重明显小于纯种波尔山羊繁殖的公羔羊初生重。经胚胎移植的难产发生率较高,达7.5%。     

河南省荷斯坦牛冷冻胚胎移植效果分析

本研究对河南省鼎元种牛育种有限公司2012~2013年荷斯坦牛冷冻胚胎移植效果进行了统计分析。结果表明,不同牧场对冷冻胚胎移植妊娠率影响显著(P0.05),牧场E单个活犊需胚胎数最低(1.8枚/头),在1~6月份和11~12月份中,月份对冷冻胚胎移植效果影响不显著(P0.05)。1~2月份进行胚胎移植时妊娠率最高(61.5%),单个活犊需胚胎数最低(2.0枚/头),3~4月份胚胎移植效果最差。移植部位对冷冻胚胎移植效果影响不显著(P0.05),但右侧移植效果略高于左侧。对13~23月龄受体牛进行移植,受体牛年龄对冷冻胚胎移植妊娠率、流产率和单个活犊需胚胎数影响差异不显著(P0.05),但13~18月龄受体牛的胚胎移植流产率高于19~23月龄。     

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)。     

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。     

影响绵羊胚胎移植妊娠率的因素分析

试验旨在研究影响绵羊胚胎移植妊娠率的关键因素。试验选取64只供体绵羊,对其进行超数排卵处理,同时选取370只受体绵羊,进行同期发情处理。采集胚胎分析其质量、数量及发育阶段,观察受体卵巢黄体数对妊娠率的影响。对试验数据进行卡方检验分析。结果表明,A级桑葚胚妊娠率(72.3%)显著高于B级桑葚胚的妊娠率(60.0%),P0.05;移植1枚A级桑葚胚妊娠率(71.8%)显著低于移植2枚A级桑葚胚妊娠率(82.4%),P0.05;移植1枚B级桑葚胚妊娠率(48.2%)显著低于移植2枚B级桑葚胚妊娠率(58.3%),P0.05;胚胎发育阶段和黄体数对妊娠率无显著性影响(P0.05)。综上所述,胚胎等级、移植胚胎数是影响绵羊胚胎移植妊娠率的主要因素。     

体内、外卵母细胞对猪体细胞克隆胚胎发育潜力的影响

为探讨猪体内、外成熟卵母细胞对核移植重组胚胎发育能力的影响,试验通过激素促排获得体内成熟卵母细胞和收集废弃卵巢获取体外成熟的卵母细胞,分别构建核移植重组胚,比较其卵裂率、囊胚率及胚胎移植受孕情况。结果显示,PGC+PMSG+HCG组的平均排卵数（27.8枚/头）显著高于PGC+HCG （12.5枚/头）、PMSG+HCG （13.7枚/头）及自然发情组（11.5枚/头）（P<0.05）,体内收集到的卵母细胞,可用于构建核移植重组胚的可用卵率均达到90%以上,与其他处理组差异不显著（P>0.05）,说明通过激素处理可获得更多的可用卵母细胞,而且卵母细胞的质量没有显著差异;以体内和体外成熟卵母细胞作为核移植受体构建的克隆胚胎,二者的胚胎融合率（80.31%和79.29%）和卵裂率（90.40%和86.51%）差异均不显著（P>0.05）,但来自体内成熟卵母细胞克隆的胚胎发育至囊胚期的比例显著升高（P<0.05）;将体内、外成熟卵母细胞构建的核移植重组胚分别移植代孕母猪,头平均移植30或60枚时,体内成熟卵母构建的克隆胚胎移植出生仔猪10头,而体外培养卵母细胞构建的克隆胚胎均未着床受孕,表明通过激素促排获得的卵母细胞质量更好,能显著提高克隆胚胎的囊胚率,减少胚胎移植数量,提高代孕母猪的怀孕率。     

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。     

Effect of Oocytes in vivo and in vitro on the Developmental Potential of Porcine Somatic Cell Cloned Embryos

To investigate the impact of porcine oocytes in vivo and in vitro maturation (IVM) on the development of porcine somatic cell cloned embryos,the somatic cell cloned embryos cultured in vitro and the sows were treated with hormones to collect mature oocytes in vivo,and the cleavage rate, blastocyst rate and embryo implantation were compared. The results showed that the average number of ovulation in PGC+PMSG+HCG group was significantly higher than that of PGC+HCG,PMSG+HCG and the natural estrus groups (P<0.05). The oocytes collected in vivo could be used for the construction, and the available oocytes rate reached more than 90%,and there was no significant difference among the four groups (P>0.05),which indicated that groups treated by hormone could obtain more available oocytes and the quality of oocytes was not significant different. In vivo and in vitro matured oocytes were used as nuclear transfer embryos of recombinant receptor,the fusion efficiency (80.31% and 79.29%) and cleavage rate (90.40% and 86.51%) were not significant different (P>0.05), but the proportion of in vivo matured oocytes cloned embryos developed into the blastocyst stage was significantly higher (P<0.05). The reconstructed embryos made from in vivo and in vitro matured oocytes were transplanted into surrogate sows (transferred 30 or 60 embryos),10 piglets were born in in vivo maturation of cloned embryo transfer group,while there was no implantation in in vitro maturation of cloned embryo transfer group. The results showed that high quality oocytes obtained by superovulation could significantly increase the blastocyst rate of embryos,reduce the number of embryos transferred and improve the pregnancy rate of surrogate sows.     

德系乳肉兼用型西门塔尔牛胚胎移植效果研究

[目的]建立德系乳肉兼用型西门塔尔牛群体,探索文山牛快速扩繁和生产性能提升技术。[方法]在马关县从600头文山黄牛、西本杂、安本杂和短本杂牛母牛中选择270头,用CIDR+PG法进行同期发情处理,选择黄体合格的受体,采用非手术法移植德系乳肉兼用型西门塔尔牛胚胎,在移植后60~90 d通过直肠检查法进行妊娠诊断,确定妊娠率,跟踪调查产犊情况,并测定胚胎移植所产犊牛的初生体重和主要体尺指标。[结果]受体牛同期发情处理270头,胚胎移植87头,妊娠28头,移植妊娠率32.18%;妊娠母牛中产犊17头,产犊率60.71%,产犊19头,成活16头,产犊成活率84.21%。公母犊牛平均的初生重33.00 kg,体高69.50 cm,体斜长62.19 cm,胸围72.69 cm,管围13.50 cm。[结论]首次在文山州开展了牛胚胎移植获得成功,杂交牛受体的移植妊娠率极显著或显著高于文山牛受体,秋季移植的妊娠率极显著高于春季和冬季的。提示以杂交牛为受体在秋季进行胚胎移植可有效提高移植成功率。     

纯种和牛快速扩繁关键技术研究

为达到快速扩繁纯种和牛的目的,试验从供体牛高强度超数排卵、胚胎性别鉴定及不同品种受体牛移植对产犊、妊娠期和犊牛初生重的影响进行研究。结果表明,纯种和牛连续超排9次,每次间隔30 d,第2次头均回收胚数22.78枚,显著高于第5~9次结果（P< 0.05）,其余各组之间无显著差异（P> 0.05）;第1~3次头均可用胚数分别为8.67、13.78、8.56枚,组间无显著差异（P> 0.05）,第2次头均可用胚数显著高于第4~9次头均可用胚数（P< 0.05）,极显著高于第7、9次头均可用胚数（P< 0.01）。性别鉴定胚胎和常规胚胎移植妊娠率和产犊率差异均不显著（P> 0.05）,性别鉴定胚胎的母犊率为95.56%。西门塔尔杂交牛受体产犊率高于和牛杂交牛、荷斯坦奶牛受体,但差异不显著（P> 0.05）。和牛杂交牛受体妊娠期极显著高于西门塔尔杂交牛和荷斯坦奶牛受体（P< 0.01）,西门塔尔杂交牛和荷斯坦奶牛之间差异不显著（P> 0.05）;西门塔尔杂交牛受体所产和牛母犊初生重显著高于和牛杂交牛受体（P< 0.05）,极显著高于荷斯坦奶牛受体（P< 0.01）,3个品种受体牛产和牛公犊初生重之间差异不显著（P> 0.05）;同一品种受体牛其产公、母犊牛的妊娠期及初生重均相近。     

Effect of mSOF and G1.1/G2.2 Media on the Developmental Competence of SCNT‐Derived Bovine Embryos

The objective of this study was to compare the effect of two culture media: modified synthetic oviductal fluid (mSOF) and G1.2/G2.2, on the developmental competence of bovine somatic cell–cloned embryos. Cloned embryos were produced by transferring adult skin fibroblasts into enucleated MII oocytes. After activation, the reconstructed embryos were randomly allotted to either mSOF or G1.2/G2.2 for culture (the embryos were transferred from G1.2 to G2.2 on days 3 of culture). The development competence of cloned embryos in these two culture systems was compared in terms of cleavage rate, blastocyst formation rate and apoptosis cell number in day 7 blastocyts. To investigate the in vivo developmental competence of cloned embryos in the two culture systems, a total of 87 and 104 blastocysts derived from mSOF and G1.2/G2.2 medium groups were transferred individually to recipient Angus cows, respectively. No differences were observed in terms of cleavage rate, day 7 blastocyst rate and blastocyst cell number between these two culture systems. However, the day 6 blastocyst formation rate was significantly higher in G1.2/G2.2 than that in mSOF. In addition, blastocysts cultured in mSOF have a higher percentage of apoptotic blastomeres compared to those in G1.2/G2.2 (8.5 ± 1.2 vs 16.8 ± 1.5, p < 0.05). Although difference in pregnancy rate was not observed 40 days after embryo transfer, significantly higher pregnancy rate was observed in G1.2/G2.2 group after 90 days of embryo transfer (12.4% vs 37.5%, p < 0.05). Moreover, calving rate was significantly improved in G1.2/G2.2 group compared to mSOF group (27.9% vs 6.7%, p < 0.05). In conclusion, our results indicate that G1.2/G2.2 can improve developmental competence of bovine SCNT embryos both in vitro and in vivo, which is more suitable for culture of bovine SCNT embryos than mSOF medium.     

胚胎质量和发育阶段对奶牛胚胎移植妊娠率的影响

[目的]为了评估胚胎质量和发育阶段对奶牛胚胎移植妊娠率的影响。[方法]使用63头青年奶牛作为供体进行超数排卵,评估回收胚胎质量和发育阶段。选择334头青年奶牛作为受体鲜胚移植不同质量和发育阶段胚胎。对胚胎质量分布、发育阶段分布、不同质量胚胎和不同发育阶段胚胎移植30 d妊娠率进行统计分析。[结果]可用胚胎中A级胚胎比例(60.78%)显著高于B级和C级胚胎比例(36.70%和2.52%)(P<0.05);致密桑椹胚比例(54.36%)显著高于早期囊胚,囊胚和扩张囊胚比例(18.35%,25.0%和2.29%)(P<0.05)。A级和B级胚胎移植30 d妊娠率(63.55%和64.35%)显著高于C级胚胎移植30 d妊娠率(44.44%)(P<0.05);致密桑椹胚、早期囊胚、囊胚和扩张囊胚移植30 d妊娠率差异不显著(P<0.05),早期囊胚、囊胚移植30 d妊娠率高于致密桑椹胚、扩张囊胚移植30 d妊娠率(P<0.05)。[结论]选择不同发育阶段的A级和B级胚胎能够获得较高胚胎移植妊娠率,增加早期囊胚和囊胚阶段胚胎移植数量能够提高胚胎移植妊娠率。     

受体绵羊人工授精后移植胚胎的试验

试验通过使用人工授精后绵羊作为受体进行胚胎移植,以寻找一种更加有效的方法提高绵羊胚胎移植的经济效益。试验中使用FSH对10只无角道赛特绵羊进行超数排卵处理,同时对60只受体小尾寒羊进行同期发情。供体羊在发情配种后4.5～5.0d从子宫角收集胚胎。同时,将胚胎移植到同期发情并进行人工授精的受体羊子宫内。总共有57枚可用胚移植给44只受体小尾寒羊,32只怀孕到分娩,共产下羔羊51只(无角道赛特羔28只,道赛特与小尾寒羊杂种羔23只)。此外,经人工授精但未进行手术移植的7只小尾寒羊产下15只杂种羔羊。移胚植受体妊娠率72.7%(32/44),移胚受体繁殖率118%(51/44),受体利用率88.3%(53/60)。移胚受体总妊娠率和受体利用率均显著高于常规ET组(P<0.01)。与常规胚胎移植相比,受体羊人工授精后移植胚胎不仅提高了无角道赛特母羊的繁殖率,而且提高了受体羊的利用率。     

季节和体重对肉用绵羊超数排卵的影响

本试验选择64只(次)健康经产的德国美利奴、无角道赛特、特克塞尔肉用绵羊,集中饲养,分别在秋季、冬季和春季,根据体重分组进行超数排卵,即41～50 kg组、51～60 kg组、61～70kg组和71～80kg组.结果表明,不同品种超数排卵的结果无显著差异(P＞0.05).不同季节对肉用绵羊超数排卵影响差异不显著(P＞0.05),但胚胎可用率秋季和冬季高于春季(P＜0.01).不同体重组对肉用绵羊超数排卵反应亦没有影响(P＞0.05),但41～50kg组和51～60kg组的可用胚数高于71～80kg组(P＜0.05),胚胎可用率41～50kg组高于其它组(P＜0.01),51～60 kg组的胚胎可用率高于71～80kg组(P＜0.01),61～70kg组的胚胎可用率高于71～80 kg组(P＜0.05).本试验结果说明肉用绵羊在秋冬春季超数排卵效果差异不明显,可在这些季节进行胚胎生产和移植.体重在41～70 kg的供体母羊进行超排效果较好.     

Use of insulin-like growth factor-I during embryo culture and treatment of recipients with gonadotropin-releasing hormone to increase pregnancy rates following the transfer of in vitro-produced embryos to heat-stressed,lactating cows

An experiment was conducted to determine whether pregnancy rates following the transfer of in vitro-produced embryos to heat-stressed cows could be improved by 1) culturing embryos in the presence of IGF-I and 2) treating recipients with GnRH. Lactating Holstein cows (n = 260) were synchronized using a timed ovulation protocol. Embryos were produced in vitro and cultured with or without 100 ng/mL of IGF-I. On d 7 after anticipated ovulation (d 0), a single embryo was transferred to all recipients with a palpable corpus luteum (n = 210). A subset of recipients (n = 164) was injected with either GnRH or placebo on d 11. Plasma progesterone concentrations on d 0 and 7 were used to determine the synchrony of recipients. Pregnancy was diagnosed at d 53 and 81 by rectal palpation. Among all recipients, transfer of IGF-I-treated embryos increased pregnancy rate at d 53 (P < 0.05) and tended to increase pregnancy rate at d 81 (P < 0.06). Calving rate also tended to be higher for recipients that received IGF-I-treated embryos (P < 0.07). Among the subset of synchronized recipients (n = 190), pregnancy rate at d 53 and d 81 and calving rate were higher (P < 0.05) for IGF-I-treated embryos. The GnRH tended to increase pregnancy rate at d 53 for all recipients (P < 0.08) and the subset of synchronized recipients (P < 0.10). There were no effects of GnRH (P > 0.10) for pregnancy rate at d 81 and calving rate. The overall proportion of male calves was 64.3%. There was no effect (P > 0.10) of embryo treatment or GnRH on the birth weight or sex ratio of calves. Results of this experiment indicate that treatment of embryos with IGF-I can improve pregnancy and calving rates following transfer of in vitro-produced embryos. Further research is necessary to determine whether the treatment of recipients with GnRH is a practical approach to increase pregnancy rates following in vitro embryo transfer.