Some Factors Affecting the Rate of Pregnancy after Embryo Transfer Derived from the Brazilian Jumper Horse Breed

This study aimed to evaluate the effects of certain embryo transfer parameters on the pregnancy rate after equine embryo transfer of the Brazilian Jumper Horse breed. The size, embryonic development stage, embryo quality, and synchronization of ovulation between the donor (n = 120) and recipient (n = 420) were evaluated in 396 embryos. Embryo recovery was performed on Day 6-9 after ovulation (Day 0 = day of ovulation). The recipient mares were chosen on the day of embryo recovery, and the transfers were performed that same day. The embryo size (diameter including envelopes; n = 396) ranged from 150 to 3000 μm; 67.1% measured between 400 and 1199 μm. The embryo size (400-1199 μm vs. ≤399 μm); stage of development (n = 396; blastocyst and expanded blastocyst versus compact morula and early blastocyst); quality (n = 396; grade 1 [excellent]), 2 [good], or 3 [poor]); and synchronization of ovulation between the donor and recipient (0, 1, 2, 3, and 4 days versus −1, 5, and 6 days, respectively) all affected pregnancy rate (P < .05). The pregnancy rate did not differ significantly among transfers performed on Days 0, 1, 2, 3, and 4. In conclusion, embryos measuring 400-1199 μm produced higher pregnancy rates in recipients than embryos measuring 150-399 μm, and blastocysts and expanded blastocysts produced pregnancy more efficiently than morulae and early blastocysts. The embryo quality also affected the pregnancy rate. Synchronization of donor and recipient ovulation to Days 0-4 improved the efficiency of embryo transplant.     

Effect of Maternal Age on the Ratio of Cleavage and Mitochondrial DNA Copy Number in Early Developmental Stage Bovine Embryos

Age-associated deterioration in both the quality and quantity of mitochondria occurs in older women. The main aim of this study was to examine the effect of age on mitochondrial DNA copy number (mtDNA number) in early developmental stage bovine embryos as well as the dynamics of mtDNA number during early embryo development. Real-time PCR was used to determine mtDNA number. In vitro-produced embryos 48 h after insemination derived from Japanese black cows, ranging in age from 25 to 209 months were categorized based on their cleavage status. There was an overall negative relationship between the age of the cow and cleavage status, to the extent that the ratio of embryos cleaved over the 4-cell stage was greater in younger cows. The mtDNA number did not differ among the cleaved status of embryos. In the next experiment, oocytes collected from each donor cow were divided into 2 groups containing 10 oocytes each, in order to compare the mtDNA number of mature oocytes and early developmental stage embryos within individuals. Upon comparing the mtDNA number between oocytes at the M2 stage and early developmental stage 48 h post insemination, mtDNA number was found to decrease in most cows, but was found to increase in some cows. In conclusion, age affects the cleaving ability of oocytes, and very old cows (> 180 months) tend to have lower mtDNA numbers in their oocytes. The change in mtDNA number during early development varied among individual cows, although overall, it showed a tendency to decrease.     

Reactive oxygen and nitrogen species regulate porcine embryo development during pre-implantation period: A mini-review

Significant porcine embryonic loss occurs during conceptus morphological elongation and attachment from d 10 to 20 of pregnancy, which directly decreases the reproductive efficiency of sows. A successful establishment of pregnancy mainly depends on the endometrium receptivity, embryo quality, and utero-placental microenvironment, which requires complex cross-talk between the conceptus and uterus. The understanding of the molecular mechanism regulating the uterine-conceptus communication during porcine conceptus elongation and attachment has developed in the past decades. Reactive oxygen and nitrogen species, which are intracellular reactive metabolites that regulate cell fate decisions and alter their biological functions, have recently reportedly been involved in porcine conceptus elongation and attachment. This mini-review will mainly focus on the recent researches about the role of reactive oxygen and nitrogen species in regulating porcine embryo development during the pre-implantation period.     

Advances in Holding and Cryopreservation of Equine Oocytes and Embryos

Methods for holding of oocytes and embryos during shipment as well as for their cryopreservation can greatly aid equine reproductive management. Oocytes can be held at room temperature overnight or at cooler temperatures for two nights without affecting maturation or embryo development after intracytoplasmic sperm injection. In contrast, methods for cryopreservation of equine oocytes that support high rates of embryo development have not yet been established. Equine embryos may be held overnight at temperatures from 5°C to 19°C without reduction in viability, but longer holding periods, or higher holding temperatures, may be detrimental. Small equine embryos (<300 μm), either in vivo derived or in vitro produced, can be slow frozen or vitrified successfully. In the last decade, methods have been developed to allow in vivo–derived expanded blastocysts, up to Day 8, to be vitrified successfully after blastocoele collapse. These methods of shipment and preservation allow mare owners in remote locations to have access to sophisticated assisted reproductive technologies.     

牛胚胎移植产业化超数排卵的系统研究

为了提高供体牛的利用率,加速胚胎移植技术的产业化,试验系统地研究了超排方案、青年奶牛的月龄及发情周期不同阶段、重复超排次数、超排间隔时间、卵巢状态、胎次与产后间隔时间对超排效果的影响,共超排供体奶牛1 127头次,回收胚胎共10 900枚,可用胚胎数量为6 942枚,平均每头次获得胚胎数量为6.73枚.结果表明:①应用PSO1、PSO2和 PSO3三种方案进行超排处理,PSO3方案的超排成功率和平均回收可用胚胎数均高于其他两个方案;②利用发情周期9～11 d的自然发情奶牛直接超排效果最好;③对青年奶牛进行超排处理,应限制超排起始时间,15月龄以上的青年奶牛超排效果较好;④供体牛连续重复超排控制在3次以内,连续4次超排处理后极大降低胚胎可用率;⑤超排处理的间隔最短时间应该选择46～60 d;⑥经产牛1～3胎次超排效果较好,7胎次以上较差;⑦产后间隔时间选取80～90 d.     

中西药对影响奶牛胚胎移植效率的繁殖疾病疗效的对比

为了提高供体牛的利用率,加速胚胎移植技术的推广和应用,试验对比了中药和西药对几种影响奶牛胚胎移植效率的繁殖疾病的疗效.结果表明:①激素疗法中GnRH一次治疗卵巢囊肿的治愈率为83.0%,GnRH PG治愈率为86.4%,中药内服一个疗程治愈率为73.8%,GnRH 中药内服治愈率为87.4%;各治疗组治愈后第一情期受胎率分别为52.3%、54.4%、58.1%和55.3%.②黄体退化不全形成硬肿的供体奶牛经2个疗程内服中药治疗,总治愈率91.3%,治愈母牛再次用于超排,所获胚胎数量及可利用率与正常母牛没有显著差异.对持久黄体的治疗以促孕灌注液和激素疗法中PG注射效果最好,治愈率分别为85.9%和89.1%;治愈后2个情期受胎率分别为77.6%和75.5%.③卵巢机能减退4个治疗组中,FSH LH组、PMsG hCG组、促孕灌注液组和中药内服组的治愈率分别为75.6%、63.8%、73.8%和82.5%;治愈后2个情期受胎率分别为62.9%、58.2%、65.7%和67.3%.④治疗慢性子宫内膜炎以土霉素油剂和促孕灌注液效果较好,治愈率分别为85.4%和84.8%,治愈后2个情期受胎率分别为83.9%和82.0%;PG注射治愈率为76.3%,治愈后2个情期受胎率为78.9%;中药内服组治愈率为79.4%,治愈后2个情期受胎率为84.7%.⑤为预防胚胎死亡采取3种控制措施,对照组、P4补充组、hCG组和内服中药组人工授精后的受胎率分别为53.3%、56.1%、62.1%和58.3%;胚胎移植后受胎率分别为43.3%、49.2%、53.3%和55.3%.治疗组的受胎率均有不同程度的提高.     

毛百合×有斑百合种间杂种的育成

以毛百合(Lilium dauricum)为母本、有斑百合(L.concolor var.prlchellum)为父本进行常规杂交,通过胚培养成功获得了种间远缘杂交种,对种问杂交亲和性、胚培养和杂种的形态特征进行了调查研究.结果表明:毛百合×有斑百合蒴果膨大率达46.1%,种子有胚率为1.30%,胚培养萌发率为15.2%;4个杂交后代生长旺盛,株高表现出较明显的超亲现象,其他性状呈趋中趋势,杂种植株的花色更偏向橙色.4个杂交后代虽然在花色上与亲本有一定差异,但其它性状均具有双亲的特征,从形态上能证明是真杂种.     

陇东马牙枣幼胚培养

以陇东马牙枣盛花后50～60d的幼胚为试材,采用L9(3)4正交设计,筛选适宜的幼胚萌发培养基。结果表明,基本培养基、激素配比、蔗糖的含量对幼胚萌发都有显著的影响;其中MS-H培养基的萌发率最高为47.62%;ZT最适宜含量为0.5mg/L,IAA最适宜含量为0.6mg/L;较高的蔗糖含量(50g/L)有利于胚的萌发。幼胚萌发的适宜培养基为MS-H+ZT0.5mg/L+IAA0.6mg/L+LH0.5g/L+蔗糖50g/L+琼脂4.0g/L,剥去种皮,幼胚萌发效果较好。当胚萌发后,将胚从胚乳中或愈伤组织中剥离继续诱导愈伤组织,再经诱导不定芽,生根获得正常植株。     

磨盘柿的胚囊发育及败育特性研究

为揭示母本育性对磨盘柿杂种败育的影响程度,本研究对磨盘柿的子房、胚囊进行定期调查和制片观察,结果表明:磨盘柿的胚囊发育进程与其子房的外观大小之间有着严格的对应关系;在胚囊发育形成过程中存在多种异常现象,主要表现为无胚囊、胚囊伸长但无扩大、局部珠被发育异常、发育滞后、胚囊的退化等,其中珠孔部的外珠被发育异常是磨盘柿所特有的,但考虑到孢子体与育性的可能关系,认为胚囊珠被局部异常与后期突起败育型杂种形成之间的关联度还需要更进一步的试验证据。