绵羊体外受精技术条件优化的研究

探讨了卵泡液、受精时间和季节对绵羊卵母细胞体外培养效果的影响。从屠宰场采集羊卵巢,抽取卵巢表面2mm-6mm的卵泡卵母细胞,进行体外成熟培养;卵母细胞成熟后,与处理后的精子共同培养进行受精。结果表明,在卵母细胞成熟培养液中添加绵羊卵泡液（SFF）组的卵裂率和囊胚发育率均优于添加同等浓度的牛卵泡液（BFF）组;受精22、18、12h组的卵裂率分别为64.85%、54.39%、48.72%,受精22h和18h组间差异显著（P〈0.05）,受精22h组和12h组间差异极显著（P〈0.01）。秋季绵羊体外受精的卵裂率和囊胚率均高于夏季,差异显著（P〈0.05）,但秋季与春季相比差异不显著（P〉0.05）。因此,进行绵羊卵母细胞体外成熟培养时,培养液中添加SFF;受精时,精卵共培养时间为22h的效果较好。在绵羊的繁殖季节秋季进行体外受精较春季和夏季的结果好。     

Ovarian follicle development and genital tract characteristics in different birthweight gilts at 150 days of age

In the last decades, selection for improved prolificacy has resulted in higher litter sizes and has thereby increased the proportion of low birthweight (LW) piglets. It is well documented that LW piglets have lower growth performance, muscle accretion and poor carcass quality. However, little is known about the relations of birthweight with subsequent reproductive performance in gilts. This study investigated the effects of birthweight on reproductive tract and ovarian follicle development in 150‐day‐old gilts. Twenty eight female pigs of different birthweight ranges (high‐HW: 1.8–2.2 kg; low‐LW: 0.8–1.2 kg) from higher parity commercial sows were reared until 150 days of age, and their body weights were recorded at weaning, end of nursery and end of the grower‐finisher phase. The animals were killed and their reproductive tracts collected for biometrical and histomorphometrical analysis. LW gilts showed significantly lower body weights and growth rates during all phases of production compared to their HW counterparts (p < .01). Most biometrical measurements of the reproductive tract were similar between the experimental groups, except vaginal length and the gonadossomatic index (relative ovarian weight), which were affected by birthweight class (p < .05). LW females also showed fewer medium size (3–5 mm; p < .01) ovarian follicles, pre‐antral follicles (p < .07) and more atretic follicles per ovarian cortex area (p < .05). Therefore, besides the effects on post‐natal growth performance, birthweight affects vaginal length and the follicular dynamics process, which may impair the reproductive performance of replacement gilts.     

Effect of aglepristone (RU534) administration during follicular phase on progesterone,estradiol-17β and LH serum concentrations in bitches

Aglepristone was administered in bitches during the follicular phase to evaluate its effects on progesterone, estradiol-17β and LH serum concentrations. Ten German Shepherds were divided into two groups (treated n = 5; control n = 5). Treated bitches received 10 mg/kg BW of aglepristone subcutaneously during the early follicular phase, 24 hr after and then 7 days later. The control group was injected, at the same time periods, with saline solution (0.3 ml/kg BW). For the steroid evaluations, blood was collected daily from the onset of proestrus until the first day of cytological dioestrus. For LH base-line serum determination, blood was also collected every 20 min for 2 hr at the onset of proestrus. For LH surge identification, blood was collected daily (every 6 hr) starting from the day of the first administration of aglepristone or saline solution until the first day of dioestrus. All animals ovulated but the treated group presented longer ovulation-dioestrus intervals than the control group (5.2 ± 2.2 days p < .05). Serum concentrations of the evaluated hormones were similar between experimental animals except for serum LH. Indeed, no LH peaks were detected in the treated group while LH surges were clearly observed in the control group (9 ± 1 days after the beginning of proestrus. In particular, the area under the curve for LH was significantly lower in treated than control animals (12 ± 4 ng/ml x Day; p = .01). In conclusion, administrations of aglepristone during the follicular phase of the bitch does not affect the steroid hormone patterns but does prevent the occurrence of a LH surge. This work raises significant questions and opens perspectives concerning the mechanisms of ovulation in bitches.     

BFF,rbGH对牛卵泡卵母细胞体外受精后发育的影响

利用屠宰黄牛卵巢，对２～５ｍｍ卵泡卵母细胞的体外成熟（ＩＶＭ）、体外受精（ＩＶＦ）、受精卵的体外培养（ＩＶＣ）进行了系列研究。结果表明，在成熟培养液中单独添加１０％Ｄ０ＥＣＳ（发情当天牛血清）获得了卵泡卵母细胞受精后较高的卵裂率（６４．７％）、桑椹胚发育率（３９．９％）和囊胚发育率（２４．８％），说明在成熟培养液中单独添加Ｄ０ＥＣＳ是可行的；在含１０％Ｄ０ＥＣＳ的成熟培养液中再添加１０％或２０％ＢＦＦ（牛卵泡液），均能提高受精卵的卵裂率以及桑椹胚和囊胚的发育率，以添加２０％ＢＦＦ效果较好，其囊胚发育率（３５．０％）显著高于对照组（２４．８％，Ｐ＜０．０５）；在卵泡卵母细胞成熟培养系统和受精卵共同培养系统中添加１０μｇ／Ｌ重组牛生长因子（ｒｂＧＨ），虽对体外受精卵的卵裂率无显著影响（Ｐ＞０．０５），但能显著提高卵裂胚的囊胚发育率（Ｐ＜０．０５）     

理囊散对不孕奶牛血清中微量元素和白介素影响的研究

本试验旨在探讨经理囊散治疗不孕症奶牛前后,血清中微量元素(Cu、Fe、Mn、Zn)与白介素(IL-1、IL-6)的含量变化规律。利用火焰法原子吸收光谱法和酶联免疫吸附法分别检测试验组和对照组治疗前、后血清中微量元素和白介素的含量。试验结果表明,①理囊散治疗卵泡囊肿有效率为83.33%,排卵延迟有效率为90.91%;②在卵泡囊肿型奶牛中,Mn、Zn含量治疗前均显著低于正常组(P<0.05),Fe治疗前显著高于正常组(P<0.05),经治疗后逐渐达到正常值水平,Cu、IL-1、IL-6含量治疗前后差异不显著;③在排卵延迟型奶牛中,Fe治疗前显著高于正常组(P<0.05),Mn含量治疗前极显著低于正常组(P<0.01),Zn含量治疗前显著低于正常组(P<0.05),经治疗后逐渐达到正常值水平,Cu、IL-1、IL-6含量治疗前后差异不显著。因此,理囊散对奶牛不孕症血清中部分微量元素和白介素含量有微调作用,在奶牛不孕症治疗效果上有不可忽视的作用。     

蛋白质营养对动物卵泡发育的影响及其作用机制

卵泡的发育对于雌性动物繁殖性能具有重要影响.蛋白质是机体酶、激素合成以及组织器官构建的原料,对于卵泡发育必不可少.蛋白质营养状态能通过肝脏雌激素受体(ER)影响胰岛素样生长因子Ⅰ (IGF-Ⅰ)、瘦素(leptin)、胰岛素(insulin)等代谢信号,进而传递至下丘脑-垂体-性腺(HPG)轴,对卵泡发育进行调控.本文综述了蛋白质营养对卵泡发育的影响及其作用机制.     

山羊卵泡卵母细胞体外受精及受精卵的体外发育

对山羊卵泡卵母细胞的体外成熟、体外受精及受精卵体外培养的系列研究表明,在成熟培养液中添加１５μｇ／Ｌ重组牛生长激素（ｒｂＧＨ）,与ＦＳＨ和ＬＨ配合使用,卵泡卵母细胞的成熟率（６７．９％）显著高于不含ｒｂＧＨ的培养组（５３．１％,Ｐ＜０．０５）;３８．５℃和３９．０℃的培养温度对卵泡卵母细胞体外成熟培养的影响无显著差异（Ｐ＞０．０５）;ＢＯ液上浮法对山羊冷冻精子的活精子分离效果优于Ｐｅｒｃｏｌ分层液法,并得到较高的受精率（６４．３％∶４９．０％,Ｐ＜０．０５）;颗粒细胞单层和输卵管上皮细胞单层的共同培养体系均有益于受精卵的体外发育。     

Distribution of immunoreactive von Willebrand factor in the microvascular network of bovine cystic follicles

The present study was undertaken to examine whether the von Willebrand factor (vWF) expression in the follicular microvasculature in the cystic follicles differs from that in the atretic follicles. Paraffin sections of healthy, atretic and cystic follicles were immunostained with rabbit monoclonal antibody to vWF. The vWF-positive cells were counted in four different regions of a follicle from the apical to the basal side. In all types of follicles, immunoreactions for vWF were observed in the endothelial cells of capillaries as well as veins and arteries in the theca interna and externa. In the theca interna, vWF-positive areas were significantly lower in the Type A and B cystic follicles compared to advanced and late atretic follicles. In the theca externa, the vWF-positive blood vessels and vWF-positive area were significantly smaller in all types of cystic follicles than in the healthy or atretic follicles. From these results, it is suggested that in the cystic follicles the induction of vWF in the follicular microvasculature system is reduced, which may suppress the degeneration of vascular system. Continuation of stability in vasculature may be one of the factors that delays the tissue regression in the cystic follicles, and also contributes to the accumulation of follicular fluid that originates from the serum.     

Increased melanomacrophage centres in the liver of reproductively dysfunctional female greater amberjack Seriola dumerili (Risso, 1810)

The greater amberjack Seriola dumerili is a new aquaculture fish that may display reproductive dysfunctions. During extensive follicular atresia, which is a common reproductive dysfunction in females during vitellogenesis, part of the reabsorbed yolk returns to the liver to be metabolized and recycled. Melanomacrophage centres (MMCs) are aggregates of macrophage-like cells that play a role in the destruction, detoxification and recycling of endogenous and exogenous materials, and have been associated with systemic stress. Wild and captive-reared greater amberjack were sampled in the Mediterranean Sea during two different phases of the reproductive cycle. The liver of reproductively dysfunctional captive-reared females sampled during the spawning season showed a high density of both MMCs and apoptotic cells. A weak liver anti-cytochrome P450 monooxygenase 1A immunoreactivity was observed, suggesting that the examined fish were not exposed to environmental pollutants. We propose that the observed increase in MMCs and apoptosis in captive-reared fish was related to the hepatic overload associated to the metabolism of yolk proteins reabsorbed during extensive follicular atresia. Since follicular atresia is a frequent physiological and pathological event in teleosts, we suggest that the reproductive state should be always assessed when MMCs are used as markers of exposure to stress or pollutants.     

卵泡液中总蛋白含量与卵泡直径的关系

通过检测不同直径卵泡的卵泡液中总蛋白含量的变化,从而进一步阐明卵泡液中蛋白成分在卵泡发育过程中的作用。将直径为3~5、5~8、8~10mm的卵泡和囊肿卵泡(>21mm)分为4组(n=10),分别抽取卵泡液。通过TCA/丙酮方法进行样品处理;根据Bradford法检测原理,利用分光光度计对其进行蛋白定量研究。结果表明:3~5mm为4440μg/mL,5~8mm为976μg/mL,8~10mm为686μg/mL,囊肿卵泡(>21mm)为537.2μg/mL。经SPSS13.0软件分析,3~5mm卵泡液中蛋白平均浓度高于其他组(P<0.01);其他3组间差异均不显著(P>0.05);囊肿卵泡中蛋白含量最低。由此可得出,卵泡液中总蛋白浓度随着卵泡直径的增加而降低,表明卵泡液中的蛋白含量与卵泡发育的成熟度呈负相关。而囊肿卵泡的发生很可能和蛋白含量降低有关,为卵泡囊肿发病机制的研究提供新的理论参考。