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1.
《当代畜牧》2007,(5)
采集中国大白兔的桑椹胚共545枚,采用OPS法,配制VSⅠ和VSⅡ两种冷冻液,分别冷冻兔的桑椹胚265枚和280枚,解冻后胚胎回收率分别为93.6%(248/265)和92.8%(260/280),两者差异不显著(P>0.05);解冻后胚胎形态良好率分别为95.2%(236/248)和94.2%(245/260),两者差异显著(P<0.05)。将解冻后形态良好的210枚桑椹胚分别移植到15只受体母兔体内,结果有8只母兔妊娠,妊娠率为53.3%(8/15),妊娠受体内移植胚胎数为110枚,产仔数占移植胚胎数的33.6%(37/110)。用VSⅠ冷冻的100枚桑椹胚移植到7只受体母兔体内,妊娠率为57.1%(4/7),产仔率为35.7%(20/56);用VSⅡ冷冻的110枚桑椹胚移植到8只受体母兔体内,妊娠率为50.0%(4/8),产仔率为31.5%(17/54)。无论妊娠率还是产仔率,VSⅠ都高于VSⅡ,但差异均不显著(P>0.05)。 相似文献
2.
绵羊体外受精卵冷冻解冻后的移植 总被引：1，自引：0，他引：1
以含有10％乙二醇的磷酸缓冲液对体外受精的绵羊桑椹胚及囊胚进行防冻和冷冻处理后保存于液氮中。解冻后将胚胎分别以去除防冻剂后移植和连同防冻剂直接移植两种方法,将30枚桑椹胚和19枚囊胚先后以外科手术法移植到28只受体母羊子宫内。结果共有8只受胎,其中4只足月妊娠后产羔4只(♀1,♂3),另外4只分别在移植后2～3个月妊娠中断,其原因尚不清楚。解冻后连同防冻剂直接移植后的受胎率为50％(4/8),明显高于去除防冻剂后移植的结果20％(4/20),(P<0.05)。受体的发情同步化程度不同(-1～+1日)对受胎率无明显影响。 相似文献
3.
牛胚胎玻璃化超快速冷冻一步法移植试验 总被引：2，自引：0，他引：2
应用 30 %乙二醇 0 .3mol/ L 蔗糖 - m- PBS液 (VS1)、30 %乙二醇 0 .3m ol/ L 蔗糖 5 %葡聚糖 (T- 5 0 0 ) - m-PBS液 (VS2 )、30 %乙二醇 0 .3mol/ L蔗糖 10 %葡聚糖 (T- 5 0 0 ) - m- PBS液 (VS3)玻璃化超快速冷冻奶牛胚胎 ,一步法移植。结果显示 ,VS1、VS2、VS3组玻璃化超快速冷冻奶牛胚胎解冻后的形态正常率分别为 10 0 % (2 0 / 2 0 )、95 %(19/ 2 0 )和 5 5 .6 % (5 / 9) ;培养存活率分别为 70 % (14 / 2 0 )、75 % (15 / 2 0 )和 2 2 .2 % (2 / 9) ;囊胚孵化率分别为 0、2 0 % (4/2 0 )和 0。VS1、VS2组玻璃化冷冻奶牛胚胎解冻后的形态正常率极显著地高于 VS3组 (P<0 .0 1) ;VS1、VS2组玻璃化冷冻奶牛胚胎解冻后的培养存活率分别显著 (P<0 .0 5 )和极显著 (P<0 .0 1)地高于 VS3组。 VS1组冷冻的胚胎经一步法移植了 5头 (1枚 /头 ) ,未获得妊娠 (0 / 5 ) ;VS2组冷冻的胚胎用一步法移植了 10头 (1枚 /头 ) ,结果获得 2头妊娠(2 / 10 ) ,并产下 2头正常犊牛。 相似文献
4.
采用2种不同浓度的冷冻液(VSⅠ和VSⅡ),通过OPS法玻璃化冷冻兔的胚胎。结果表明:冷冻兔的桑椹胚641枚,解冻后胚胎总回收率为93.8%(601/641),形态结构良好率为94.5%(568/601),透明带完好率为100%(601/601)。VSⅠ和VSⅡ冷冻的胚胎解冻后回收率分别为93.8%(305/325)和93.7%(296/316),两者差异不显著(P>0.05);形态良好率分别为95.1%(290/305)和94.0%(278/296),前者显著地高于后者(P<0.05)。将解冻后形态结构良好的桑椹胚415枚移植到29只受体母兔体内,总妊娠率为51.7%(15/29),产仔率为34.5%(61/177)。VSⅠ妊娠率为53.3%(8/15),产仔率为36.8%(35/95);VSⅡ妊娠率为50.0%(7/14),产仔率为31.7%(26/82)。无论妊娠率还是产仔率,VSⅠ都高于VSⅡ,但差异不显著(P>0.05)。 相似文献
5.
《中国畜牧杂志》2016,(3)
本实验旨在研究猪孤雌激活4-细胞胚胎的冷冻保存效果。胚胎采用Cryotop法进行玻璃化冷冻保存,解冻后分析其存活率、胞内活性氧(ROS)和谷胱甘肽(GSH)水平、囊胚发育率及囊胚质量。结果表明:冷冻胚胎体外恢复2 h的存活率与新鲜胚胎无明显差异(100%vs.96.8%,P0.05),但当其培养时间达到24 h时,存活率显著下降至88.3%(P0.05)。另外,玻璃化冷冻导致胚胎内ROS水平显著升高(P0.05),GSH水平显著下降(P0.05)。与新鲜胚胎相比,冷冻胚胎获得的囊胚发育率明显降低(59.8%vs.26.4%,P0.05),但囊胚的凋亡细胞率、内细胞团数、滋养层细胞数及总细胞数均无明显差异(P0.05)。结果显示,玻璃化冷冻猪孤雌激活4-细胞胚胎导致其存活、氧化还原能力和囊胚发育下降,但仍能获得较高质量的囊胚。 相似文献
6.
胚胎分割技术在肉牛繁育中的研究与应用 总被引：1，自引：0，他引：1
本试验采用弗莱维赫肉牛和日本和牛作为供体,本地黄牛为受体,进行同期发情和超数排卵,共得到可用胚胎234枚,分为4个处理:新鲜全胚113枚、新鲜双半胚55枚、冷冻全胚56枚和冷冻双半胚10枚,进行移植。结果表明,新鲜双半胚的妊娠率为65.45%,略高于新鲜全胚移植的妊娠率(63.72%),差异不显著(P＞0.05)。冷冻双半胚产犊率为20.00%,低于冷冻全胚移植的妊娠率(39.29%),差异不显著(P＞0.05),而冷冻组的妊娠率均显著低于新鲜组(P＜0.05)。新鲜双半胚的产犊率为95.56%,双犊率为35.55%,犊牛存活率为72.09%。新鲜全胚组的产犊率为63.16%,显著低于新鲜双半胚组(P＜0.05),但犊牛存活率却显著高于新鲜半胚组(90.00%和72.09%)(P＜0.05)。此外,全胚与双半胚移植后产生的犊牛初生重之间并无显著差异(P＞0.05)。 相似文献
7.
探讨了F1代(♀ICR×♂BALB/C6J)和ICR小鼠体外受精及体内受精两种不同来源获得的8-细胞胚胎玻璃化冷冻效果。不同来源的小鼠胚胎在预热到37℃1MDMSO溶液中处理后,转移到冷冻管中5min然后加入DAP213,并在0℃平衡5min后冷冻。体外受精及体内受精两种方法获得的F1代小鼠8-细胞胚胎解冻后的存活率分别为92.5%和93.33%;ICR代小鼠8-细胞胚胎解冻后的存活率分别为65.83%和67.50%。同一品系体外受精与体内受精来源的8-细胞胚胎冷冻复苏后胚胎存活率差异不显著(P>0.05),但不同品系体外受精及体内受精来源的8-细胞胚胎复苏率差异均显著(P<0.05),表明8-细胞胚胎冷冻复苏率的高低与胚胎获得的方法无关,但与品系的不同有关。 相似文献
8.
不同冷冻和解冻方法对小鼠桑椹胚发育的影响 总被引：1，自引：0，他引：1
本试验以2种程序化冷冻液和2种玻璃化冷冻液对昆明白系小鼠的桑椹胚进行细管法冷冻保存,比较程序化冷冻-管外解冻和玻璃化冷冻-管内解冻对胚胎体内、外发育的影响。胚胎体外培养结果表明:玻璃化冷冻组及程序化冷冻组胚胎发育率(95.3%￣95.8%,98.9%)无显著(P>0.05)差异。将程序化冷冻、EFS30玻璃化冷冻以及新鲜的胚胎各168枚移植给假孕受体鼠,妊娠受体产活仔率各组间相比(50.8%,58.3%,54.9%)无显著性(P>0.05)差异。结果证明,玻璃化冷冻保存的胚胎管内解冻效果好,为生产中家畜的胚胎移植提供了理论和技术参考。 相似文献
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10.
本研究采用不同来源的体外生产胚胎(屠宰场卵巢IVF胚胎,OPU-IVF胚胎,SCNT胚胎),系统研究了不同冷冻方法对水牛体外生产胚胎冷冻效果的影响,以完善水牛体外生产胚胎冷冻方法,进一步提高胚胎冷冻效果。试验选用6～7日龄囊胚分别用不同的冷冻液和不同冷冻方法进行胚胎冷冻。玻璃化冷冻液分别为40%EG、25%EG+25%DMSO和20%EG+20%DMSO+0.5 mol/L蔗糖;程序化冷冻液分别为10%甘油和0.05 mol/L海藻糖+1.8 mol/L EG+0.4%BSA。结果表明:(1)在玻璃化冷冻中,无论何种胚胎不同冷冻液的冷冻效果有明显的差异,但均以20%EG+20%DMSO+0.5 mol/L蔗糖作为冷冻液的冷冻效果最好,并且高于程序化冷冻的存活率;而对于程序化冷冻,用10%甘油作为冷冻液,屠宰场卵巢IVF胚胎的冷冻后存活率略高于用0.05 mol/L海藻糖+1.8 mol/L EG+0.4%BSA的冷冻后存活率,但差异不显著(P>0.05)。(2)VF胚胎利用程序化冷冻胚胎解冻后,在0～24 h内有76.5%胚胎复活,高于玻璃化冷冻的复苏率(48.9%)(P<0.05);而与此相反,在24～48 h内,玻璃化冷冻胚胎的复苏率(42.6%)则高于程序化冷冻(23.5%)(P<0.05)。综上所述,各种来源的水牛体外生产胚胎均可进行冷冻保存,应用玻璃化冷冻的效果好于程序化冷冻,且以20%EG+20%DMSO+0.5 mol/L蔗糖作为冷冻液进行玻璃化冷冻效果最好,但程序化冷冻后的胚胎复苏速度明显快于玻璃化冷冻的速度。 相似文献
11.
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。 相似文献
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B Alexander G Coppola GF Mastromonaco E St. John ER Reyes DH Betts WA King 《Reproduction in domestic animals》2008,43(2):207-211
Early pregnancy diagnosis and monitoring play an important role following embryo transfer in sheep. The aims of the current study were to investigate (i) the pattern of serum progesterone profiles in sheep carrying somatic cell nuclear transfer (SCNT)‐derived (clone) pregnancies, and (ii) the frequency of pregnancy loss during development following SCNT embryo transfer. Sheep SCNT embryos were made using standard nuclear transfer techniques. Day 7 embryos were surgically transferred to oestrus‐synchronized recipients (n = 27). As a control, normal fertile ewes (n = 12) were bred by natural breeding. Serum was collected from all the ewes on the day of estrus (day 0 sample), 7 days post‐estrus (day 7 sample) and 19 days post‐estrus (day 19 sample) and every 10 days thereafter until lambing or pregnancy loss occurred. Serum progesterone (P4) was assessed using enzyme immunoassay. Pregnancy was confirmed by ultrasound scanning on day 35 of pregnancy followed by subsequent scanning every 10 days. In control ewes, pregnancy rate on day 35 was 83.3% (10/12), whereas in the ewes that received SCNT embryos, it was 22.2% (6/27; p < 0.05). The day 45 pregnancy rate in the control ewes was 83.3%, whereas in the SCNT embryo recipients it was 11.0% (p < 0.05). Hormone analysis revealed that SCNT embryo recipients exhibited a significantly lower P4 profiles at different time points in pregnancy compared to controls (p < 0.05). This study highlights the use of serum progesterone in combination with ultrasound for the investigation of embryo loss and crucial times during development of normal and SCNT embryos in sheep. Further, the serum P4 levels directly reflect the degree of placental development in these two groups. 相似文献
13.
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。 相似文献
14.
在胚胎移植生产中,用从新西兰引进的肉用绵羊品种无角陶赛特羊作供体进行规模化胚胎移植生产.超排结果表明:两次超排分别回收平均卵数为5.62枚和9.08枚;从不同年龄的无角陶赛特羊上获得总卵数差异不显著;配种后第7天冲取胚胎,以桑椹胚为最多,占总回收胚胎数的75.78%,配种后第8天回收的胚胎,以扩展囊胚和孵出囊胚数所占比例为最多,分别占总数的25.89%和31.75%;从两侧卵巢所观察到的黄体数和所回收的胚胎数差异不显著;用人工授精法比试验中的其它方法能显著提高所获得的有效胚胎数;用促卵泡素恒量注射、孕马血清两次注射法进行超排效果好,所获得的平均胚胎数为8.53枚,有效胚占回收总胚的百分率则增加到82.11%. 相似文献
15.
The effect of boar exposure during artificial insemination (AI) on semen backflow, fertilization, and embryo quality was evaluated. Gilts (approximately 170 d) were induced into estrus with PG600, and ovulation was synchronized using hCG 72 h later. Estrus detection was initiated after PG600 and continued at 12-h intervals. At estrus, gilts were allotted to receive boar exposure (BE, n = 20) or no boar exposure (NBE, n = 20) during AI. Gilts receiving NBE were identified to be in estrus prior to AI and the boar was then removed for 1 h, whereas gilts in the BE group received 15 min of exposure during AI. Insemination occurred in crates at 12 and 24 h after onset of estrus with 3 x 10(9) sperm/80 mL. Backflow was collected continuously with samples taken at time 0, (during AI), and at 0.25, 0.5, 0.75, 1, 2, 4, and 8 h after first and second AI. The effect of treatment was evaluated for time of insemination (min), backflow (mL), and sperm in backflow samples. Oviducts were flushed 2 d after first AI to evaluate the effect oftreatment on fertilization rate, accessory sperm numbers on embryos (scored 1 to 5), and embryo quality. There was no effect of first or second AI; therefore, data were pooled. Average duration of AI was 3.7 +/- 0.2 min and was not influenced by BE (P < 0.10). However, during the initial stage of AI, BE reduced the volume of semen (18.6 vs 32.4 +/- 3 mL) and the number of sperm lost (0.8 vs 1.3 +/- 0.15 x 10(9) sperm) compared to NBE (P < 0.05). There was a treatment x time effect (P < 0.05) for volume of backflow. By 45 min, the BE gilts lost more volume (9.0 vs 3.6 mL) compared to the NBE group, but sperm loss did not differ. Between 1 and 8 h after AI, neither volume nor sperm loss was influenced by treatment. By 8 h, total leakage (65 vs 63 mL) and total sperm loss (1.6 x 10(9) vs 1.8 x 10(9) sperm) were not influenced by BE (P > 0.10). However, more accessory sperm (P < 0.01) were found on embryos for the NBE (> or = 11 sperm/embryo) compared to BE embryos (< or = 10 sperm/embryo). Despite this observation, percentages of fertilized embryos (99.5 +/- 0.5 %) and number of embryos (11.5 +/- 0.1) were not different (P > 0.10). In conclusion, AI in the presence of a mature boar did not affect total semen leakage, sperm loss, fertilized embryos, or embryo quality. The importance of boar exposure during insemination was evident from less leakage during insemination, but had no effect on fertility; this suggests that the elimination of boar exposure during AI may not be deleterious to reproductive performance. 相似文献
16.
The objective of this study was to investigate the effects of beta‐mercaptoethanol (β‐ME) on post‐thaw embryo developmental competence and implantation rate of mouse pronuclear (PN) embryos that were cryopreserved after slow freezing, solid surface vitrification (SSV) or open‐pulled straw (OPS) vitrification methods. Mouse PN embryos were cryopreserved by using slow freezing, SSV and OPS methods. After cryopreservation, freeze–thawed PN embryos were cultured up to blastocyst stage in a defined medium supplemented without or with 50 μm β‐ME. The blastocyst formation rate of embryos that were cryopreserved by slow freezing method (40.0%) or vitrified by OPS method (18.3%) were lower than those vitrified by SSV method (55.6%) and fresh embryos (61.9%) in the absence of 50 β‐ME in the culture media (p < 0.05). The blastocyst formation rate of embryos that were cryopreserved by slow freezing method (53.1%) or by OPS method (41.9%) were lower than those vitrified by SSV method (79.5%) and that of fresh (85.7%) in the presence of β‐ME in the culture media (p < 0.05). The embryos transfer results revealed that the implantation rate of blastocyst derived from mouse PN embryos vitrified by SSV method (31.9% vs 51.2%) was similar to that of the control (39.0% vs 52.5%), but higher than those cryopreserved by slow freezing (28.2% vs 52.0%) and by OPS method (0.0% vs 51.2%) (p < 0.05). In conclusion, supplementation of β‐ME in an in vitro culture medium was shown to increase survival of embryo development and implantation rate of frozen–thawed mouse PN embryos after different cryopreservation protocols. 相似文献
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18.
RE Green BFS Santos CC Sicherle FC Landim-Alvarenga SD Bicudo 《Reproduction in domestic animals》2009,44(3):406-410
The aim of this study was to evaluate the viability in the effect of open pulled straw (OPS) vitrification procedure of sheep embryos after direct transference. Embryos were produced in vivo and cryopreserved in slow freezing or OPS vitrification. The survival rates of cryopreserved embryos were compared to non-frozen standard pattern. In a first set of experiments, embryos at morula and blastocyst stages were dived in ethylene glycol (1.5 M) and frozen in an automatic freezer. After being thawed, they were directly or indirectly transferred to ewes recipient. A second group of embryos were drawn into OPS and plunged into liquid nitrogen after being exposed at room temperature for 1 min and 45 s in 10% EG plus 10% dimethyl sulphoxide (DMSO), then again for 30 s in 20% EG + 20% DMSO + 0.5 M sucrose. After being warmed, embryos were also directly transferred using a French mini straw as the catheter for the transplantation process or after in vitro dilution of cryoprotectants (two-step-process). No significant difference was observed among fresh, frozen or vitrified embryos on pregnancy rate (50.0%, 38.6% and 55.8%). However, when we evaluated only the direct transference, the pregnancy rate of OPS vitrified embryos was higher than that of frozen embryos (57.1% vs 34.8%) (p = 0.07). In addition, vitrified morulae had a higher pregnancy rate than the one with frozen embryos (64.0% vs 38.9%) (p = 0.07). Finally, our results indicate that OPS vitrification technique in association with direct transference improves the viability of sheep embryos with potential applications to field conditions. 相似文献
19.
从美国引进高产奶牛性控雌性胚胎，在农村条件下，选择自然发情的黄牛作受体，移植合格受体牛648头。结果显示，自然发情黄牛924头中，经直肠检查合格受体牛648头，使用合格率为70．13％（648／924）；氯前列烯醇同期处理476头，使用合格率为36．13％（172／476），差异极显著（P〈0．01）。80～100d怀孕检查，自然发情移植成功率41．98％（272／648），同期发情移植成功率40．12％（69／172），差异不显著（P〉0．05）。自然发情黄牛不同黄体级别的移植成功率分别为：A级44．09％（41／93），B级42．43％（199／469），C级37．21％（32／86），差异不显著（P〉0．05）。单胚移植580头，移植成功率42．41％（242／580），双胚移植68头，移植成功率44．12％（30／68），差异不显著（P〉0．05）。正常胚胎单枚移植541头，移植成功率42．51％（230／541），裸胚单枚移植39头，移植成功率30．77％（12／39），裸胚比正常胚移植成功率低11．74％。怀孕牛已出生犊牛83头。其中母犊79头．公犊4头，母犊占95．18％，与自然发情的性比例（50％）相比较，差异极显著（P〈0．01）。 相似文献