幼龄羊的周龄及卵母细胞体外培养体系对JIVET技术的影响

为了探讨周龄及卵母细胞体外培养体系对幼龄羊胚胎体外生产技术(JIVET)的影响,试验对10只5~10周龄杜寒杂种羔羊进行激素诱导,获取卵母细胞,分别在基础成熟液和含200μmol/L半胱氨酸的成熟液里成熟,再分别在含2%发情羊血清(ESS)的Ⅰ受精液和含3 mg/m L牛血清白蛋白(BSA)的Ⅱ受精液里受精,测定不同周龄羔羊的采卵数量、卵母细胞成熟率和卵裂率。结果表明:5,7,8,10周龄羔羊获得卵母细胞数分别为(111.00±16.97)枚/只、(139.50±28.99)枚/只、(108.50±17.68)枚/只、(42.00±11.31)枚/只,5,7,8周龄羔羊显著高于10周龄羔羊(P0.05),而5,7,8周龄羔羊之间差异不显著(P0.05);试验成熟液获得的卵母细胞成熟率比基础成熟液高3.64百分点;Ⅰ受精液获得的卵母细胞卵裂率极显著高于Ⅱ受精液(P0.01)。说明选择5~8周龄羔羊、卵母细胞体外成熟液添加一定量半胱氨酸和体外受精液含一定量ESS可以改善JIVET体系。     

辽宁绒山羊母羔体外胚胎生产技术研究

以辽宁绒山羊母羔为研究对象,通过超排母羔、卵母细胞体外成熟、体外受精、胚胎移植技术等试验研究,建立了绒山羊母羔体外胚胎生产技术体系。结果表明:PMSG+FHS组与FSH+PMSG组超排方法获得的卵母细胞数无显著差异(P0.05),但PMSG+FHS组卵裂率和囊胚率均极显著高于FSH+PMSG组(P0.01)。成熟液添加EGF组PBⅠ率、卵裂率均显著高于对照组(P0.05);20 ng/m L EGF组PBⅠ率显著高于10 ng/m L组(P0.05),卵裂率差异不显著(P0.05)。卵母细胞成熟培养24~27 h组第一极体率与卵裂率均显著高于21~24 h组(P0.05);SOF受精体系卵裂率显著高于TALP(P0.05),囊胚率极显著高于TALP受精系(P0.01)。秋季胚胎移植受体母羊16只,产羔3只。     

特克赛尔羊JIVET技术研究

探讨特克赛尔羊JIVET(juvenile in vitro embryo technology)技术,为优种绵羊JIVET技术体系完善提供技术支撑。试验对4~8周龄特克赛尔羊超数排卵,采用活体采卵的方法及对卵母细胞的体外成熟—受精—受精卵培养—受精卵移植的方法进行特克赛尔羊JIVET技术研究。结果显示:4只4~8周龄特克赛尔羊获得可用卵母细胞296枚,平均74枚/只;获得2~4细胞期受精卵205枚,卵裂率为69.26%,平均获2~4细胞期受精卵51.25枚/只;将66枚2~4细胞期受精卵移植给11只受体羊,6只羊受孕,生产11只羔羊,受胎率为54.55%。研究表明,本试验方法可作为特克赛尔羊JIVET技术研究的方法。     

幼畜胚胎体外生产技术的研究进展

家畜JIVET技术采用外源促性腺激素处理幼龄母畜,诱导其卵泡大量发育,并与活体采卵、卵母细胞体外成熟、体外受精技术相结合体外生产胚胎,经胚胎移植后产生后代。本文综述了JIVET技术的研究现状,以及JIVET技术中幼畜的选择、超排方案、卵母细胞采集和体外培养的条件等,并进一步分析了JIVET技术生产胚胎的影响因素及存在的问题。     

幼畜胚胎体外生产技术的研究进展

家畜JIVET技术采用外源促性腺激素处理幼龄母畜,诱导其卵泡大量发育,并与活体采卵、卵母细胞体外成熟、体外受精技术相结合体外生产胚胎,经胚胎移植后产生后代.本文综述了JIVET技术的研究现状,以及JIVET技术中幼畜的选择、超排方案、卵母细胞采集和体外培养的条件等,并进一步分析了JIVET技术生产胚胎的影响因素及存在的问题.     

幼龄杜泊羊胚胎体外生产技术的研究

为探讨幼龄杜泊羊胚胎体外生产技术(JIVET),给幼龄杜泊羊卵巢卵泡的利用提供技术支撑,试验采用促卵泡素(FSH)和孕马血清(PMSG)对4~8周龄杜泊羊进行诱导、活体采集卵母细胞、卵母细胞体外成熟-受精-受精卵培养及2~4细胞期受精卵移植研究。结果表明:从8只4~8周龄杜泊羊中采集到可用卵母细胞585枚,平均73.13枚/只;获得2~4细胞期受精卵346枚,卵裂率59.15%;将77枚2~4细胞期体外受精卵移植于17只受体母羊的输卵管壶腹部,7只母羊受孕,产羔13只,受胎率41.18%。说明采用4~8周龄杜泊羊胚胎体外生产技术可作为JIVET的有效技术。     

不同超数排卵方法所获羔山羊卵母细胞体外成熟率及体外受精率的比较

本研究观察了不同超排处理方法对羔山羊卵母细胞体外成熟及体外受精的影响，并将体外受精胚胎进行了移植，以便探讨利用羔山羊卵母细胞生产体外受精胚胎的技术途径。对２～４月龄羔山羊用ＦＳＨ、ＦＳＨ＋ＬＨ（静脉注射）和ＦＳＨ＋ＬＨ（肌肉注射）三种方法进行超排处理，平均分别回收１６．０、２０．３和１５．０枚卵母细胞。将这些卵母细胞进行２０～２４ｈ的成熟培养后，其成熟率分别为４５．５％，４４．４％和７６．５％。成熟卵母细胞经体外受精，体外发育培养后其２～４细胞胚胎的发生率分别为５．６％ａ、６．７％和３１．７％ｂ（ｂ，ｖｓ，ａ，Ｐ＜０．０５）。将９枚２～４细胞期胚胎移植给６只受体母羊，结果有２只妊娠并产羔２只。     

幼畜胚胎体外生产技术研究进展

家畜JIVET技术采用外源促性腺激素处理幼龄母畜,诱导其卵泡大量发育,并与活体采卵、卵母细胞体外成熟、体外受精技术相结合体外生产胚胎,经胚胎移植后产生后代。本文综述了JIVET技术的研究现状、基本操作步骤,并进一步分析了JIVET技术中的影响因素,以及该技术所存在的问题和发展前景。     

不同周龄酉州乌羊幼羔超排及卵母细胞体外发育研究

选择4~8、8~12、12~16周龄羔羊进行超排处理,研究羔羊卵巢与卵泡的发育情况、卵母细胞活体采集效果及其体外发育能力。结果表明:羔羊超排处理后卵巢大小随着周龄的增加明显下降;4~8周龄组可用卵泡数、回收卵母细胞数极显著高于8~12周龄组和12~16周龄组(P0.01),但卵母细胞的回收率随着周龄的增加而升高;12~16周龄的A、B级卵母细胞所占比率较高,A、B级与C级卵母细胞的成熟率差异极显著(P0.01);对成熟卵母细胞进行体外受精,C级卵母细胞卵裂率显著低于A级与B级卵母细胞(P0.05);酉州乌羊幼羔体外胚胎移植妊娠率为27.03%,生产羔羊14只。结果提示,4~8周龄酉州乌羊羔羊超排效果优于8~12、12~16周龄段羔羊,活体采集的羔羊卵母细胞可经体外成熟、体外受精和胚胎移植获得后代。     

牛羊繁育提速技术在京成功 绵羊繁育效率比常规胚胎移植提高20倍

从澳大利亚引进的新一代牛羊快速繁育技术-牛羊幼畜超排技术(JIVET技术),在北京创新科农农牧科技有限公司香河基地试验成功。这一技术将大大加速我国牛羊育种进程和优良种畜的扩繁。JIVET技术是集羔羊的超数排卵、卵母细胞体外成熟、体外受精、胚胎体外培养和胚胎移植等技     

Developmental Capabilities of Embryos Produced In Vitro from Prepubertal Lamb Oocytes

Breeding from prepubertal females, known as juvenile in vitro embryo transfer (JIVET), reduces the generation interval and increases the rate of genetic gain in animal breeding programs. While the birth of the first lambs from prepubertal ewes occurred nearly 30 years ago; and there is considerable interest in the commercialization of this technology, its efficiency remains too low. The advent of in vitro production (IVP) of embryo resulted in the more widespread use of JIVET. Morphologic and metabolic differences coupled with reduced in vitro and in vivo development of oocytes derived from prepubertal animals have been reported. Research has been undertaken to optimize donor selection and hormone stimulation methods in an attempt to reduce the variability and increase the proportion of donors responding to hormone stimulation and increase oocyte developmental competence. Yet, this variation persists and the development of oocytes and embryos from prepubertal animals remains reduced when compared with adults. Recent improvements to JIVET, resulting from a modified hormone stimulation regime, have eliminated the failure of donors to respond to hormone stimulation, and increased both the number and developmental competence of oocytes harvested from very young prepubertal lambs. This increased efficiency has facilitated the incorporation of other reproductive technologies such as sperm sexing with JIVET, resulting in the birth of lambs of a pre-determined sex from prepubertal lambs. Increased rates of genetic gain in sheep breeding programs can be achieved by combining sexed sperm with oocytes obtained from lambs as young as 3–4 weeks of age. Continued increases in the efficiency of JIVET resulting from further improvements to hormone stimulation regimes and an increased understanding of the differences between oocytes from adult and prepubertal animals will result in the commercialization of this technology.     

羔羊诱导卵泡发育及其应用的研究进展

幼畜体外胚胎移植(Juvenile In Vitro Embryo Transfer,JIVET)技术是利用幼畜对外源激素敏感的生理特点,采用外源促性腺素诱导幼畜卵泡超数发育,结合卵母细胞体外成熟、体外受精和胚胎移植等技术生产后代,该技术体系的研究与应用可以充分发挥优秀母畜繁殖潜能,快速扩繁良种畜群。近年来,国内外对绵羊JIVET技术研究比较多,但应用JIVET技术生产体外胚胎的效率低下、效果不稳定仍然是一个普遍问题。迄今为止,大多数的研究致力于加强供体羔羊选择、优化激素处理方案以及提高羔羊卵母细胞体外发育能力等方面。本文综述了羊JIVET技术的原理与最新研究进展以及影响JIVET技术效率所存在的内因和外因,旨在为深层次探索羔羊卵子发生和卵泡发育调控机制提供理论依据,促进JIVET技术的研究与应用。     

羔羊超数排卵与胚胎体外生产技术研究进展

自从幼畜体外胚胎移植(JIVET)技术问世以来,因其在优秀种畜快繁、育种世代间隔缩短、遗传资源保护等方面具有广阔的应用前景,引起了很多科研机构的关注.为提高JIVET的生产效率,研究人员对该技术的多个环节进行了探索,但总体效率还不是很高.论文就羔羊超数排卵与胚胎体外生产技术实施中的关键环节,即羔羊超排供体的年龄、超排季...     

无角道赛特和萨福克肉羊胚胎移植研究

引进无角道赛特和萨福克肉羊,利用胚胎移植技术,加快良种畜群的纯种繁育进程。阜新市肉羊研发中心利用4年时间共超排供体羊99只,采集可用胚胎883枚,利用手术法移植受体羊850只,受胎558只,胚胎移植受胎率达到63．2%。研究结果表明：胚胎移植技术应用于畜牧生产进入商品化推广阶段是可行的。     

萨福克羔羊0~65日龄生长发育规律研究

试验对萨福克羔羊0~65日龄生长发育规律进行了分析,旨在完善肉羊的生长发育规律研究,为萨福克羔羊0~65日龄的饲养管理及营养水平提供理论依据。试验于新疆华兴牧场惠康畜牧种羊繁育基地进行,供体母羊为纯种萨福克羊,受体母羊为哈萨克羊,并对出生的萨福克羔羊进行体尺、体重测量和记录,将数据输入Excel表格进行整理,并通过SPSS 16.0软件分析各性状平均值和各个体尺指标与体重平均值的相关性。结果表明,0~30日龄萨福克羊生长发育速度相对较快,公羔臀宽与体重的相关系数为0.989;管围与体重的相关系数为0.638;母羔臀宽与体重的相关系数为0.981,管围与体重的相关系数为0.654。0~30日龄萨福克羊公羔各性状的生长发育比母羔快,45~65日龄生长速度一致。羔羊0~15日龄体高、体长、胸围和体重等生长速度最快,而30~65日龄体高、体长、体重等部位生长速度较平稳;由体尺、体重相关性分析可知,臀宽与体重相关系数最大,管围和体重相关系数最小。由研究可知,0~30日龄营养要求关键,公羔与母羔的生长发育无明显差异,欲提高羔羊的生长发育速度则要注意羔羊各年龄阶段的生长发育规律,合理提供营养并提高饲料质量与管理水平。今后对萨福克羔羊的饲料管理中应该注重不同时期的饲养配方,合理改善饲养环境,使其发挥最大生长潜能。     

幼羔胚胎体外生产技术研究进展

幼羔胚胎体外生产技术(JIVET)是全球草食类动物繁殖技术中最先进的生物工程技术。国内外对幼羔经激素诱导后生产胚胎已作了许多研究,并取得一定的成果,本文就其研究进展作一综述。     

Optimizing female allocation to reproductive technologies considering merit,inbreeding and cost in nucleus breeding programmes with genomic selection

Female reproductive technologies such as multiple ovulation and embryo transfer (MOET) and juvenile in vitro fertilization and embryo transfer (JIVET) have been shown to accelerate genetic gain by increasing selection intensity and decreasing generation interval. Genomic selection (GS) increases the accuracy of selection of young candidates which can further accelerate genetic gain. Optimal contribution selection (OCS) is an effective method of keeping the rate of inbreeding at a sustainable level while increasing genetic merit. OCS could also be used to selectively and optimally allocate reproductive technologies in mate selection while accounting for their cost. This study uses stochastic simulation to simulate breeding programmes that use a combination of artificial insemination (AI) or natural mating (N), MOET and JIVET with GS. OCS was used to restrict inbreeding to 1.0% increase per generation and also to optimize use of reproductive technologies, considering their effect on genetic gain as well as their cost. Two Australian sheep breeding objectives were used as an example to illustrate the methodology—a terminal sire breeding objective (A) and a dual‐purpose self‐replacing breeding objective (B). The objective function used for optimization considered genetic merit, constrained inbreeding and cost of technologies where costs were offset by a premium paid to the seedstock breeder investing in female reproductive technologies. The premium was based on the cumulative discounted expression of genetic merit in the progeny of a commercial tier in the breeding programme multiplied by the proportion of that benefit received by the breeder. With breeding objective B, the highest premium of 64% paid to the breeder resulted in the highest allocation of reproductive technologies (4%–10% for MOET and 19%–54% for JIVET) and hence the highest annual genetic gain. Conversely, breeding objective A, which had a lower dollar value of the breeding objective and a maximum of 5% mating types for JIVET and zero for MOET were optimal, even when highest premiums were paid. This study highlights that the level of investment in breeding technologies to accelerate genetic gain depends on the investment of genetic improvement returned to the breeder per index point gain achieved. It also demonstrates that breeding programmes can be optimized including allocation of reproductive technologies at the individual animal level. Accounting for revenue to the breeder and cost of the technologies can facilitate more practical decision support for beef and sheep breeders.     

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。     

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.