青海高原地区应用绵羊胚胎移植技术的效果分析

采用同期发情+超排对12只特克塞尔羊、无角陶赛特羊进行处理;同时采用自然发情+超排对12只德国肉用美利奴母羊进行处理。结果表明:采用同期发情+超排处理,每只特克塞尔母羊平均获卵胚8.7枚,获受精卵数7.1枚;陶赛特母羊平均获卵胚数1.8枚,获受精卵数1.8枚(P<0.01)。德国肉用美利奴供体母羊的平均获胚数13.5枚,可用胚8.7枚,可用胚率67.0%(P<0.01)。对85只受体绵羊进行同期发情处理,结果表明,处理后同期发情率达88.2%。将12枚可用胚胎移入11只受体,其中10只受体移入单胚,1只受体移入双胚,移植成功率达83.3%(P<0.01)。对62只自然发情的受体母羊进行胚胎移植,结果表明,移植2枚胚胎受体母羊的受胎率比移植1枚的要高34.0%(P<0.01)。     

澳洲美利奴超细毛羊超数排卵与胚胎移植试验

利用从澳大利亚进口的澳洲美利奴超细毛羊69只(其中经产母羊48只,初产母羊21只)做供体,小尾寒羊320只做受体,进行胚胎移植试验,结果表明:经产母羊的发情率、采胚数、有效胚胎数和可用率(有效胚胎数/采胚数)均高于初产羊.69只处理供体羊有66只发情,收集到441枚有效利用胚胎,320只处理受体羊,有265只可用做胚胎移植,178只小尾寒羊产羔,产羔265只,受胎率和有效胚胎利用率分别为67.2%和60.1%,平均每1.7枚胚胎产1只羔羊,平均每只供体羊可获羔羊4只.     

杜泊绵羊胚胎移植技术在哈密地区应用效果观察

2014年在新疆哈密地区伊吾县喀尔里克公司进行绵羊鲜胚移植。受体羊采用两次注射PG法同期发情处理本地哈萨克母羊460只,发情389只,同期发情率为84.5%。供体羊采用PG与阴道栓埋置法同期发情处理,结合FSH递减法超排处理。结果显示:79只供体羊冲出可用胚胎485枚,冷冻胚胎42枚,移植可用胚胎443枚,移植受体羊375只,移植A级胚胎302枚,B、C级胚胎141枚。通过移植,A级胚胎利用率与受胎率明显高于B、C级胚胎,差异均显著(P0.05);71只羊采用PG法,8只羊CIDR法同期处理超排可用胚胎6.24枚与5.25枚,差异不显著(P0.05);供体羊平均超排可用胚胎6.1枚,移植受胎率达76.2%,羔羊成活率为95%。     

不同品种受体对绵羊胚胎移植效果及羔羊早期生长性能的影响

旨在探讨以小尾寒羊和蒙古羊作为受体时,二者的胚胎移植效果及所产羔羊的早期生长性能之间的差异。选用南非肉用美利奴羊(n=11)和澳洲白绵羊(n=110)作为供体,以小尾寒羊(n=196)和蒙古羊(n=504)作为受体;对供体母羊进行超数排卵以及人工授精处理,记录供体母羊的收集胚胎总数和可用胚胎数,计算平均每只羊收集的可用胚胎数以及胚胎合格率;对小尾寒羊和蒙古羊进行同期发情及胚胎移植处理,记录受体母羊的产羔数,计算繁殖率;计算并比较不同受体母羊所产羔羊的平均初生重、成活率以及70日龄断奶羔羊数、70日龄断奶重、平均日增重、平均每只母羊提供的断奶羔羊数。结果表明,从供体母羊中共获得可用胚胎549枚,平均每只羊收集可用胚胎4.54枚,胚胎合格率为82.56%;利用胚胎移植技术,移植受体母羊529只,所产羔羊240只,平均繁殖率为45.37%;小尾寒羊的发情率和繁殖率明显高于蒙古羊;共获得70日龄断奶羔羊211只,羔羊成活率为87.92%,平均日增重为280.00g;小尾寒羊和蒙古羊所产的羔羊初生重没有显著差异(P>0.05),小尾寒羊所产羔羊的70日龄断奶重和平均日增重分别极显著(P<0.01)和显著(P<0.05)高于蒙古羊,而蒙古羊所产羔羊的成活率显著高于小尾寒羊(P<0.05);平均每只小尾寒羊母羊可提供的断奶羔羊数(0.42只)略高于蒙古羊(0.39只),小尾寒羊作为胚胎移植受体略具有优势。综合分析表明,在内蒙古兴安盟地区小尾寒羊和蒙古羊均可以作为胚胎移植的受体来使用。     

季节对杜泊羊体内胚胎生产与移植效果的影响

为了研究季节对杜泊羊体内胚胎生产效率及胚胎移植妊娠率的影响,于2016年选取内蒙古赛诺草原羊业有限公司种羊场经产纯种杜泊母羊作为供体,采用同期发情、超数排卵及腹腔镜人工输精等方法对供体羊进行处理后,利用手术冲胚的方法获得纯种杜泊羊胚胎,同时对受体羊进行同期发情处理和胚胎移植。经统计分析,2016年全年该公司共有放栓供体4 241只,超排处理4 180只,配种供体4 131只,冲胚供体3 987只,供体可用率为94.01%;冲出胚胎总数为23 516枚,可用胚胎18 002枚,胚胎可利用率为76.55%,平均每只供体能获得可用胚胎4.52枚。供体在10月、11月、12月冲胚所获得的平均冲胚数和平均可用胚数明显高于4月、5月、6月(P<0.05),其中10月的平均冲胚数和平均可用胚数最高(P<0.01),分别为7.04枚和5.62枚。将获得的部分胚胎用于胚胎移植,移植单胚怀孕受体数为4 446只,妊娠率为53.67%;移植双胚怀孕受体数805只,妊娠率65.02%。此外,受体在1月、3月、6月、12月受胎率较其他月份高(P<0.05)。该研究可为杜泊种羊超数排卵、胚胎移植和肉羊的产业化生产提供一定的数据参考。     

青海高原地区应用绵羊胚胎移植技术的效果分析

采用同期发情+超排对12只特克塞尔羊、无角陶赛特羊,采用自然发情+超排对12只德国肉用美利奴母羊进行处理。结果表明:采用同期发情+超排处理,平均每只特克塞尔母羊获卵达8.7枚,平均获受精卵数7.1枚;陶赛特母羊平均1.8枚,受精卵数7枚,平均获受精卵数1.8枚(P<0.01)。德国肉用美利奴羊供体母羊的平均获胚数可达13.5枚,可用胚达8.7枚,可用胚率(%)迭67.0%(P<0.01)。对85只受体绵羊进行同期发情处理结果表明,处理后同期发情率达88.2%。将12枚可用胚胎移入10只受体,其中11只受体移入单胚,1只受体移入双胚,移植成功率达63.3%(P<0.01)。对62只自然发情的受体母羊进行胚胎移植,结果表明,移植2枚胚胎,受体母羊的受胎率比移植1枚的要高34%(P<0.01)。同期发情时间差异为±0d,妊娠率达100%,而时间差异为±0.5d的为66.67%(P<0.01)。对母羊发情时间与超排效果的关系分析结果表明,撤栓后24h内,或为注射FSH后第1天,标准时间内排卵(≤±4h)的母羊占64%,延时8 h以内排卵的占27%,提早或延时1.0h~1.5h排卵的占9%。     

波尔山羊腹腔内窥镜鲜胚移植试验

2006年10～11月在农五师兽医站种羊场使用2种超排药物对40只经产波尔山羊进行超排处理,供体羊有2只未发情,超排有效率为95%(38/40)。共回收胚胎388枚。其中,可用胚311枚,每只山羊收获胚胎10.21枚(388/38),平均可用胚8.18枚(311/38),可用胚率80.15%(311/388)。同期处理受体399只,有320只发情,同期率为80.20%,运用腹腔内窥镜技术移植受体267只,其中44只供体移植双胚。经试情150只受体未返情,受胎率为56.18%(150/267)。     

鄂尔多斯细毛羊胚胎移植效果与分析

为了解决细毛羊优质种源短缺的问题,选用鄂尔多斯细毛羊种羊作为供体羊,蒙古羊作为受体羊,进行胚胎移植试验,以期通过胚胎移植技术进行优质种源快速扩繁。试验选取55只(其中36只进行二次冲胚)鄂尔多斯细毛羊母羊作为供体,以蒙古羊作为受体,采用递减法注射FSH进行超排处理。经同期发情、超数排卵、人工授精等过程后,采用手术法子宫角回收供体的胚胎,经鉴定后移植。结果表明:91只供体共获得胚胎788枚,可利用胚胎479枚,供体平均冲胚8.66枚/只,供体平均可利用胚胎5.26枚/只,胚胎可利用率60.79%。将479枚胚胎移入469只受体,妊娠290只,受胎率为61.83%,产羔253只,繁殖率为53.94%。     

特克塞尔绵羊在山西右玉的胚胎移植效果

选取12只纯种特克塞尔羊作为供体,并采用递减法注射FSH进行超数排卵,发情后采用人工授精方式进行配种,手术法回收胚胎,鉴定后移植.164只右玉绵羊作为受体,采用两次PG注射法进行同期发情.结果表明12只供体获得胚胎共129枚,可用胚胎109枚,每只供体羊获得9.1枚可用胚胎,胚胎合格率为84.50%.134只受体表现发情,发情率为81.70%,其中同期发情的羊108只,同期发情率65.85%.109枚胚胎移入97只受体,其中12只移植双胚.61只受体被确认为妊娠,产下52只羔羊,产羔率为53.60%.12只供体手术后产下17羔羊.     

高原地区肉羊胚胎移植试验

2003年在青海省东部农业区和环湖牧区开展了以特克赛尔、萨福克和无角陶赛特为主要品种的肉羊胚胎移植试验。对农区55只小尾寒羊受体和牧区76只半细毛羊受体进行了同期处理,小尾寒羊同期发情率81·82%,半细毛羊发情率63·16%。半细毛羊移植胚胎44枚,产羔13只,流产1只(双羔),移植受胎率31·82%;每只小尾寒羊移植2枚胚胎,共移植20只受体40枚胚胎,产羔5只,其中1只母羊产双羔,受胎率12·50%。所产18只羔羊全部成活。     

胚胎移植试验羊群发情规律的研究

为了有效提高绵羊体细胞克隆和转基因等研究工作的效率,缩短胚胎移植试验羊群中空怀个体的利用周期间隔,试验对采用不同移植方法的胚胎移植受体及供体绵羊进行了移植前后的发情规律研究。结果表明:采用自制孕激素海绵栓和进口阴道孕激素释放装置(CIDR)对绵羊进行同期发情处理时发情率均接近90%,且92%以上的个体发情起始时间均集中在撤栓后24～48 h。在胚胎移植试验结束后,未参加胚胎移植个体和子宫角法未移植受体第2情期发情起始时间主要集中在上次发情后的14～20 d,而输卵管法未移植受体和输卵管法冲卵供体的发情时间则比较分散,呈现明显延后状态。未参加胚胎移植个体和子宫角法未移植受体人工授精后的受胎率接近往年羊场的平均水平,输卵管法未移植受体的受胎率略有下降,且输卵管法冲卵供体的受胎率明显降低。     

Results of the use of nonsurgical embryo transfer in the breeding of heifers

The results are analyzed of superovulation stimulation in the donors and the non-surgical embryo transfer in cattle. After superovulation induced by means of PMSG (Bioveta, Ivanovice in Haná) 6.2 and 6.3 ova and embryos were obtained, of which 66 to 70%, i. e. 4.2 and 4.3 embryos, respectively, were applicable in transfer. When the combination of PMSG with antigonadotropin (Antisergon, Bioveta, Ivanovice in Haná) was used, the embryos had somewhat better quality and their applicability was increased. After adjustment of the time of Antisergon administration the number of recovered ova and embryos was increased to 10.3 with 82.3% applicability (8.5 embryos per superovulation/donor). After super-ovulation stimulated with Folicotropin (Léciva, Praha) the yield of ova was 12.7, and 10.1 of the embryos (79.8%) were applicable. After the use of FSH-P (Burna-Biotec) 13.8 and 10.9 embryos were obtained per one superovulation. Non-surgical embryo transfer was performed in 1321 recipients over the last two years and a half. In 1984 the conception rate was 52.4%, in 1985 the average survival increased to 58.33% and in the first half of 1986 it reached 65.5%.     

牛胚胎移植技术在中国内蒙古的研究和应用

1991－1996年，对6个品种138头供体母牛超排处理199次，共获得A、B级胚胎1148枚，平均每次每头获可用胚5．77枚；共移植受体1299头（包括鲜胚和冻胚），妊娠产犊593头，总移植妊娠率为45．7％。对牛的超数排卵技术进行了深入研究，对不同超排方法的超排效果、不同品种肉牛的超排反应及重复超排的效果进行了分析，获得了适于重复应用、效果稳定的超排方案，邓用FSH－PG法，FSH总剂量在4－6mg,5d 10次递减注射。同时，对受体牛饲养管理条件与受体诱导同期发情效果和移植妊娠率之间的关系进行了探讨，对经产牛和育成牛受本移植妊娠效果进行了比较，并对移植人员技术熟练程度对妊娠率的影响进行了分析，从而为大规模实施牛ET计划提供了受体牛选择依据。     

Effects of recloning on the efficiency of production of alpha 1,3-galactosyltransferase knockout pigs

Obtaining sufficient transgenic cells via selective cultivation of genetically manipulated somatic cells is difficult due to the limited number of cell divisions. Additionally, if irreversible mutations in a cell's chromosomes occur during selective cultivation and the cell is used as the nuclear donor, somatic cell nuclear transfer (SCNT) embryos often exhibit abnormal development. On the other hand, a SCNT method in which fetal cells derived from SCNT embryos are used as the nuclear donor (recloning method) is an effective technique for obtaining large quantities of transgenic cells. In this study, we compared the in vivo development rate of SCNT embryos produced from porcine alpha1-3 galactosyltransferase gene knockout (GTKO) cells by a recloning method with that of SCNT embryos produced without recloning from porcine GTKO cells (direct method). In the direct method, 557 and 462 cloned embryos were produced using two types of activation methods, the two-step activation (TA) method and the delayed activation (DA) method, and then transferred into 6 and 4 recipients, respectively, but no piglets were born from these recipients. In the recloning method, 956 and 1038 cloned embryos were produced using the TA and DA methods, respectively, and then transferred to 8 and 7 recipients, respectively. Two piglets were born from one recipient in the TA group and 6 piglets were born from 3 recipients in the DA group. This report indicates that the recloning method improved the developmental capacity of SCNT embryos reconstructed with gene-targeted somatic cells.     

猪微注射基因导入系统影响受体受孕率及产仔率的几个因素

在应用显微注射、胚胎移植系统技术，进行猪ＯＭＴ／ＰＧＨ基因导入的研究中，对移入受体的胚数，ＰＭＳＧ处理受体以及不同的移植方法（自体移植与异体移植）等影响受体受孕率及产仔率的因素，进行了试验分析，结果表明：（１）移入受体的注射胚数分别为１０—１９枚、２０—２９枚、３０枚以上时，其受孕率为４５．５％、６４．７％和７１．４％，产仔率为８．０％、１９．６％和１４．５％，以移入２０—２９枚效率最高；（２）用ＰＭＳＧ对受体母猪做同期发情处理，其受孕率和产仔率比选择自然发情的受体分别下降２５％和８．５％；（３）采用自体移植的方式，在移入胚数基本相同的情况下（１５枚左右），比异体移植的受孕率和产仔率分别提高１４．３％和６．４％。     

Production and development of calves from sexed-bisected bovine embryos

Sixty-five chromosomal preparations from bisected bovine embryos were examined and 43 embryos (66.2%) with metaphase plates were observed; however, only 25 embryos (38.5%) were sexed. Fifteen of the bisected embryos sexed by chromosomal analysis were transferred to each of the 15 recipients, and 5 recipients became pregnant. Female calves were born on December 28, 1985, August 2, 1986 and July 18, 1988. The first female calf born from a sexed-bisected embryo (December 28, 1985) was the first case in Japan. The gestation lengths and birth weights of these calves were 277, 278 and 274 days, and 42.0, 44.0 and 37.0 kg, respectively. These two calves grew within the range of the "Standard developmental growth curve of the Holstein heifer" of the Japanese Holstein Association. The milk yield from 2 sexed-bisected cows were recorded. In the first cow (Case 1), she produced 8,575 kg of milk, 5.0% fat and 9.4% solids-non-fat (SNF) per year. In the other cow (Case 2), her expected milk volume was 7,800 kg per year. Confirmation of parentage was done by blood typing, and certified by the Japanese Livestock Animal Improvement Association.     

Controversial aspect of using GFP as a marker for the production of transgenic cattle

The objective of this study was to examine the feasibility of identification and selection of cattle embryos based on green fluorescence (GFP-positive) in order to obtain calves carrying an integrated transgene. The construct used (pbLGTNF-EGFP) contained the human tumor necrosis factor alpha (hTNFalpha) gene fused to the bovine beta-lactoglobulin promoter (bLG) in plasmid vector pCX-EGFP. In four experiments, 76 zygotes were injected; eight of them developed to the morulae/blastocysts stage of which only five were GFP positive (one of them 100%, one-50%, three- 25%). All of the GFP positive embryos were transferred to recipients. Two calves were born: one after transfer of the 100% GFP positive embryo and the other after transfer of one of the 25% GFP positive embryos. Both animals were healthy with normal weight when compared to two control calves. The integration of pbLGTNF-EGFP in the host genome could not be detected in either of the calves, suggesting that GFP is an unreliable marker for preimplantation screening of embryos.     

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。     

Use of embryo transfer to induce twinning in beef cattle: embryo survival rate, gestation length, birth weight and weaning weight of calves

Experiments were conducted in 1985 and 1986 at the Eastern Ohio Resource Development Center, Belle Valley, to examine the feasibility of using embryo transfer to induce twinning and to examine the influence of twinning on traits of the cow and calf. Embryos were collected from a total of 14 superovulated Angus donors on two dates each in 1985 and 1986 and were transferred to Angus recipients. A total of 124 embryos were transferred to 79 recipients, with 43 (34.7%) calves born alive. Seven of 45 (15.6%) recipients implanted with two embryos produced twins. In no case did both halves of the 15 embryos that were split to produce identical twins and implanted in the same recipient survive to birth. Proportion of calves born alive did not differ among transfer codes 3 (nonsplit embryos from two different donors implanted in separate uterine horns of the same recipient), 6 (nonsplit embryos from one embryo flush implanted in separate uterine horns of the same recipient) and 7 (nonsplit embryos from two different donors implanted in the same uterine horn of one recipient). Surgical transfers tended to result in a higher proportion of embryos surviving to birth (.43 vs .21; P = .16) and a higher twinning rate (.29 vs .04; P = .36) than did nonsurgical transfers. Age of recipient did not influence embryo survival (P = .98) or twinning rate (P = .99). Gestation length was 5 d shorter (P less than .01) for twin calves than for singles. Singles were 9 kg heavier (P less than .01) at birth and 32 kg heavier (P less than .01) at weaning than twins. However, cows raising twins produced 108 kg (51%) more total weaning weight than did cows raising singles.