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1.
在超声波扫描仪的指导下,用双孔型采卵针以11.3~12.0kPa抽吸压经阴道对8头高产奶牛分别连续实施7次活体采卵,每4d1次,共采集卵子340个,占可见卵泡数的53.5%.每次头均采集卵子6.1个.卵子经过体外培养、体外受精、体外受精胚体外发育培养,于体外受精后48h、168h统计的卵裂率、囊胚发育率分别为75.0%和29.7%.研究结果表明,在超声波扫描仪指导下对奶牛连续进行活体采卵,将所得卵子进行体外成熟、体外受精、体外培养,在生产中是可行的.  相似文献   

2.
奶牛活体采卵与屠宰场卵巢卵子体外生产胚胎的效果比较   总被引:1,自引:0,他引:1  
先利用屠宰场卵巢获得的卵母细胞进行体外培养试验.在建立了培养体系获得较稳定的囊胚率的基础上,再利用具有优良乳用遗传性状的老龄奶牛进行活体采卵(OPU),获得的卵子进行体外胚胎生产,对奶牛活体采卵与屠宰场卵巢卵子体外生产胚胎的效果进行了比较.同时对奶牛重复OPU的效果及OPU卵子的培养天数对妊娠率的影响因素等进行探讨,试图为研究和生产建立一个高效的活体采卵体外胚胎生产方法.结果表明:(1)活体采卵获得的卵母细胞从形态以及后期的发育情况均明显差于屠宰场卵巢获得的卵子;OPU法的卵裂率、囊胚率(70.5%、18.3%)明显低于从屠宰场卵巢获得的卵子的卵裂率、囊胚率(75.1%、34.8%)(P<0.01).(2)不同个体采集卵子数量差异明显(5.92枚/头与1.4枚/头)(P<0.05).(3)同一头奶牛每次采卵数之间无显著差异,但有波动性变化.(4)奶牛OPU卵子经培养在第6 d或第7 d发育至囊胚的胚胎移植妊娠率高于第8 d发育至囊胚的胚胎.  相似文献   

3.
《四川草原》2005,(3):39-39
奶业重大科技专项经过科研人员的攻关在奶牛性别鉴定、XY精子分离、活体采卵、体外受精等技术上取得突破。奶牛性别鉴定成功率达到98%;利用活体采卵技术,使每头供体奶牛每周可获得3~5枚卵子;建立了稳定、有效的牛卵母细胞体外受精的溶液体系和体外受精胚的体外培养技术体系,  相似文献   

4.
水牛分离精子与不同来源卵母细胞体外受精的效果研究   总被引:1,自引:1,他引:0  
本研究的目的是探讨水牛分离精子与不同来源(活体采卵或屠宰场卵巢采卵)卵母细胞体外受精的效果。活体采卵是选用20头空怀河流型母水牛(其中摩拉母牛12头,尼里-拉菲母水牛8头)每间隔3 d采卵1次,连续采卵5~6周,活体采集卵母细胞;屠宰场卵巢采卵是收集屠宰场水牛卵巢,用10 mL注射器连接18 G针头吸取水牛卵巢上可视的卵泡来收集卵母细胞。将收集的AB级水牛卵母细胞在相同的条件下进行体外成熟、然后用分离或未分离精子进行体外受精以及体外培养至囊胚。结果发现:活体采卵组和屠宰场收集的水牛卵母细胞组用分离精子受精分裂率和囊胚率没有差异(P>0.05);分离精子和未分离精子的体外受精分裂率和囊胚率也没有差异(P>0.05)。由此说明,水牛分离精子可以用于体外生产性控胚胎。  相似文献   

5.
从牛活体卵母细胞的采集方法、体外成熟培养、体外受精及早期胚胎发育培养等方面进行研究。结果表明 :(1 )供试奶牛前后两次采卵的最短间隔为 6d,采集回收卵子数最多为 2 2枚和2 1枚 ,最少为 3枚和 5枚 ;(2 ) 2头供试奶牛采集回收卵子平均为 1 0 .72± 1 1 .6枚 /次和 1 0 .5± 7.5枚 /次 ,A、B级卵子比例分别为 70 .34% (83/ 1 1 8)和 61 .88% (1 38/ 2 2 3)。(3)受精后 48h观察卵裂结果 ,分别为 45.76%和 32 .38% ,囊胚发育率分别为 2 9.66%和 2 0 .95%。  相似文献   

6.
利用超声波活体采卵技术(ovum pick-up,OPU)对12头纯种雪龙黑牛间隔4d采卵2次,对51头F2代遗传性能优秀的雪龙黑牛每周采卵1次,并对活体采卵与屠宰采卵及体外受精后胚胎的发育进行了对比分析.结果表明,活体采卵第2次卵泡数比第1次多82个,而获得的卵母细胞数第2次比第1次少1 2枚;屠宰采卵优秀卵母细胞率高于活体采卵,并且囊胚发育率高于活体采卵.  相似文献   

7.
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。  相似文献   

8.
本研究目的在于探讨利用初情期前犊牛卵子进行体外受精生产胚胎的可行性。共处理了三头母犊牛、每头用促排卵素(FSH,7.5毫克)作肌肉注射三天(六次),肌注停止即通过手术将卵巢取出,抽取卵子进行体外受精。平均每枚卵巢抽取了11.8枚卵子(71枚卵子/6个卵巢),其中有83.1%的卵子发育成熟,退化率为32.2%,受精后有52.2%卵子发生卵裂,体外培养11枚卵裂球没有一个形成囊胚。  相似文献   

9.
[目的]研究牛活体采卵-体外受精(ovum pick-up and in vitro fertilization,OPU/IVF)体系,建立高效的牛体外胚胎生产系统。[方法]以从屠宰场采集的健康牛新鲜卵巢为试验材料,进行卵母细胞体外成熟、体外受精、体外胚胎培养相关条件的摸索,重点考查体外胚胎培养液中添加瘦素(leptin)对囊胚率的影响。选取13~15月龄健康荷斯坦奶牛及和牛各10头作为供体,进行活体采卵、卵母细胞体外成熟、体外胚胎生产,记录可用卵数及可用囊胚数,统计卵裂率、囊胚率;2个品种牛的体外冷冻胚胎解冻后,以荷斯坦奶牛为受体进行胚胎移植,移植后45 d统计妊娠率。[结果]添加30 U/mL的leptin可以显著(P<0.05)提高屠宰场来源牛体外胚胎的囊胚率。随机选择供体牛活体采卵,平均每头荷斯坦奶牛获得可用卵7.5枚,平均每头和牛获得可用卵8.1枚;荷斯坦奶牛及和牛的卵裂率分别为84.00%、82.71%,二者差异不显著(P>0.05)。体外培养条件下,平均每头荷斯坦奶牛获得可用囊胚4.3枚,平均每头和牛获得可用囊胚3.7枚;荷斯坦奶牛的囊胚率(57.33%)显著...  相似文献   

10.
本研究比较了不同来源的水牛卵母细胞的体外受精及其胚胎发育。对10头摩拉母水牛连续6周进行活体采卵,共采集292枚卵母细胞,头均回收卵母细胞4.87枚,头均A级卵数为3.07枚,从屠宰场收集74头水牛卵巢共采集559枚卵母细胞,头均回收卵母细胞和A级卵母细胞分别为7.55枚和5.20枚,均显著高于活体采集的水牛卵母细胞(P<0.01)。将收集的AB级水牛卵母细胞在相同的条件下进行体外成熟、体外受精和体外培养至囊胚。结果表明,活体采卵组和屠宰水牛卵母细胞组受精分裂率差异不显著,分别为54.81%和57.73%。屠宰水牛卵母细胞组的囊胚率则高于活体采卵组(28.78%vs21.34%,P<0.05)。利用两组的胚胎进行常规法冷冻保存,冻胚解冻后的存活率差异不显著(P>0.05)。  相似文献   

11.
试验从屠宰场收集了 4 8头青年黄牛、34头青年水牛的卵巢共 16 4个 ,共回收可用卵母细胞 86 4枚。水牛平均每个卵巢可回收 3.2 2枚可用卵母细胞 ,相当于黄牛 (6 .72枚 )的一半。试验结果表明 :水牛卵巢生长卵泡少 ,卵母细胞产量少、质量差 ;3种采集黄牛卵泡卵母细胞的方法 ,用抽吸加切割法平均从每个卵巢回收的可用卵数 (7.71枚 )都极显著高于抽吸法 (6 .19枚 )和切割法 (6 .4 4枚 ) (P <0 .0 1) ,但是该法手续繁杂 ,适于一次且只能收集少量卵巢时使用 ;在黄牛和水牛卵母细胞成熟培养液中用 0 .3%PVP代替 10 %FBS ,牛卵母细胞的体外成熟率无显著差异 (P >0 .0 5 )。  相似文献   

12.
The effect of cysteamine on in vitro nuclear and cytoplasmic maturation of equine oocytes collected by transvaginal ultrasound guided follicular aspiration was assessed. Oocytes were matured in vitro with (cysteamine group) or without (control group) cysteamine. The nuclear stage after DNA Hoechst staining, penetration rates after two different in vitro fertilization (IVF) techniques (IVF media with ionophore and Hepes buffer with heparin) and the embryo yield following oocyte intra-oviductal transfer were used as a criterion for assessing nuclear and cytoplasmic maturation, respectively. Contrary to the data described in other domestic species, there was no effect of cysteamine on in vitro nuclear maturation, IVF or in vivo embryonic development under our conditions. Ovum pick up yields (52%) and maturation rates (control group: 47% and cysteamine group: 55%) were similar to those previously reported. From 57 oocytes transferred to the oviduct in each group, the number of embryos collected was 10 (17%) in the control group and five in the cysteamine group (9%). Those two percentages were not statistically different (p > 0.05). No effect of IVF technique was seen on the success rate (6%) in each group.  相似文献   

13.
在动物卵母细胞体外成熟及胚胎体外培养体系中添加一定浓度的发情牛血清可能提高卵母细胞的成熟率及胚胎的发育率。本研究以屠宰场卵巢来源的绵羊卵母细胞为试验材料,探讨了发情牛血清对绵羊卵母细胞体外成熟及孤雌胚发育的影响。结果表明,成熟液中添加10%第1天的发情牛血清能显著提高绵羊卵母细胞的体外成熟率及孤雌胚卵裂率(P<0.05);孤雌胚体外培养72 h后,向培养液中添加10% 第7天的发情牛血清能显著提高绵羊孤雌胚的桑囊胚率(P<0.05)。结果表明,发情牛血清能够促进绵羊卵母细胞的体外成熟率及孤雌胚发育率。  相似文献   

14.
This study was designed with the final goal of improving in vitro embryo production in the Thai swamp buffalo (Bubalus bubalis carabensis). Oocytes were collected by ovum pick-up (OPU) from six non-lactating multiparous swamp buffalo twice per week for 10 consecutive sessions followed by once-weekly collection for 10 consecutive sessions without hormone stimulation. In addition, oocytes were collected from slaughterhouse ovaries that were classified as follows: ovaries from non-pregnant cows with a visible corpus luteum (NPCL); pregnant cows with a corpus luteum (P); and non-pregnant cows without a corpus luteum (NP). Follicles in each group of ovaries were categorized as small (2-4 mm), medium-sized (5-8 mm) or large follicles (≥ 9 mm). The quality of the oocytes was assessed by their capacity to undergo in vitro maturation. The total number of observed follicles per session (all sizes combined) was larger in the once-weekly OPU group compared with the twice-weekly OPU group. In particular, the numbers of small and large follicles were higher in the once-weekly OPU group (5.2 ± 0.7 and 0.9 ± 0.2, respectively) than in the twice-weekly OPU group (3.9 ± 0.5 and 0.5 ± 0.1). The number of medium-sized follicles did not differ between the groups. The percentages of oocytes with an abnormal spindle morphology were not different between oocytes from the twice-weekly (30.0%) and the once-weekly (28.6%) OPU groups. A higher percentage of oocytes obtained in vitro (49.5%) exhibited nuclear abnormalities compared with those obtained in vivo (≤34.8%) after in vitro maturation. In conclusion, oocytes can be successfully collected by OPU in the swamp buffalo, without hormonal pretreatment, and per week more good-quality oocytes can be collected by twice-weekly OPU. In addition, oocytes collected from slaughterhouse ovaries can be used with the reproductive status of the cow having no influence on the maturation competence of oocytes.  相似文献   

15.
The aim of this retrospective study was to compare the number of follicles, cumulus oocyte complexes (COCs) and cultured In Vitro Produced (IVP) embryos obtained from 1396 non‐stimulated Ovum Pick‐up (OPU) sessions on 81 donor animals in a twice weekly OPU scheme. Results were obtained from 640 sessions following FSH‐LH superstimulation, on 112 donors subjected to OPU once every 2 weeks. The stimulation protocol started with the insertion of an ear implant containing 3 mg norgestomet (Crestar, Intervet, Belgium) 8 days before puncture (day ?8). The dominant follicle was ablated by ultrasound‐guided follicle puncture on day ?6. On day ?3 and day ?2, cows were injected with FSH (Ovagen, ICP) twice daily (8 am to 8 pm ), i.e. a total dose of 160 μg FSH and 40 μg LG per donor per stimulation cycle. Animals were punctured 48 h after the last FSH injection (day 0). Progesterone implants were removed the next day. Stimulated donor cows were treated with this protocol at 14‐day intervals. Follicles were visualized with a Dynamic Imaging ultrasound scanner, equipped with a 6.5 MHz sectorial probe. Follicles were punctured with 55 cm long, 18 gauge needles at an aspiration pressure corresponding to a flow rate of 15 ml/min. Cumulus oocyte complexes were recovered and processed in a routine IVF set‐up. Results demonstrate that, expressed per session, FSH stimulation prior to OPU increases production efficiency with significantly more follicles punctured and oocytes retrieved. However, when overall results during comparable 2‐week periods are considered (four non‐stimulated sessions vs one stimulated), more follicles are punctured and more oocytes are retrieved using the non‐stimulated protocol. No significant differences in the number of cultured embryos could be detected, indicating that FSH/LH stimulation prior to OPU might have a positive effect on in vitro oocyte developmental competence as more embryos are cultured with less, presumably better‐quality, oocytes.  相似文献   

16.
将不同直径牛卵泡内卵母细胞体外成熟培养22h后,对其体外受精后的卵裂率,胚胎发育速度和胚胎染色体异常发生率进行了研究。结果表明,体外受精后48h,直径3-6mm卵泡内卵母细胞外受精后的卵裂率明显高于直径1-2mm和7-10mm卵泡内卵母细胞体外受精后的卵裂率(P<0.05)。体外受精后48h,直径3-6mm包泡内卵母细胞体外受精后的胚胎的发育速度明显快于直径1-2mm和7-10mm卵泡组胚胎(P<0.05)。通过对不同卵裂期胚胎的染色体标本分析表明,直径1-2mm卵泡内卵母细胞体外受精后的胚胎染色体异常发生率明显高于直径3-6mm卵泡组(P<0.05),但与直径7-10mm卵泡组之间无显著性差异。  相似文献   

17.
In this study, we examined the effects of superstimulation using follicle‐stimulating hormone (FSH) followed by gonadotropin‐releasing hormone (GnRH) on buffalo embryo production by ultrasound‐guided ovum pick‐up (OPU) and in vitro fertilization (IVF). Nine Murrah buffaloes were subjected to OPU‐IVF without superstimulation (control). The morphologies of the oocytes collected were evaluated, and oocytes were then submitted to in vitro maturation (IVM). Two days after OPU, same nine buffaloes were treated with twice‐daily injections of FSH for 3 days for superstimulation followed by a GnRH injection. Oocytes were collected by OPU 23–24 hr after the GnRH injection and submitted to IVM (the superstimulated group). The total number of follicles, number of follicles with a diameter > 8 mm, and number of oocytes surrounded by multi‐layered cumulus cells were higher in the superstimulated group than in the control group (p ≤ 0.05). After IVF, the percentages of cleavage and development to blastocysts were higher in the superstimulated group than in the control group (p < 0.05). In conclusion, superstimulation improved the quality of oocytes and the embryo productivity of OPU‐IVF in river buffaloes.  相似文献   

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