山羊孤雌胚胎的体外培养

系统地研究了培养液微滴的大小、培养液体系及血清浓度对山羊孤雌胚胎早期体外发育的影响。结果表明，培养液微滴以较大体积(200～500μL)的培养效果较好；在3种培养液中，添加10％的NGS对山羊孤雌胚胎的体外发育效果较好，囊胚率分别可达62．79％(81／129)、53．52％(38／71)、13．64％(12／88)；mCRlaa组囊胚发育率和囊胚细胞数显著低于SOFaa组和CRlaa组，SOFaa组优于CRlaa组，尽管SOFaa组和CRlaa组间无显著差异。在本试验条件下，以SOFaa培养液，山羊孤雌胚胎体外培养72h时加入10％的NGs，500μL微滴培养，其发育效果较好，囊胚率可达62．79％。     

山羊胎期肺发育的形态学研究

对3～22周龄山羊胎儿肺进行了肉眼、光镜和透射电镜观察，结果表明：1．7～22周龄山羊胎儿肺的外部形态与胎龄无关，肺的外部形态以左二右四叶者为多见．肺的叶间裂和右肺副裂常不完整，以浆膜、肺组织或混合性组织(肺组织及浆膜)融合；2．山羊胎儿肺的发育分为5个时期：胚胎期(3～5周)肺芽分支形成主支气管，主支气管长度不断增长并萌芽出叶支气管，均衬以假复层柱状上皮。腺状期(6～12周)以支气管树发育为主，小支气管衬以假复层和／或单层柱状上皮；终蕾呈腺状，上皮细胞由假复层柱状逐渐变为单层柱状，胞核向细胞顶端移行；终蕾上皮细胞游离面可见短小的微绒毛；线粒体、粗面内质网及核糖体随着胎龄增加而逐渐增多，它们均位于细胞顶部。小管期(13～14周)以呼吸部发育为主，原始肺泡开始形成，呼吸性细支气管衬以未分化的立方上皮；终蕾腺状结构逐渐消失，终蕾上皮细胞由高矮不等的单层柱状上皮逐渐演变为立方形的原始肺泡上皮；细胞游离面可见较多的微绒毛，胞质内线粒体、粗面内质网及核糖体较发达。囊状期(第15周)呼吸部发育显著，肺内细支气管及其末端呈现出“充气”状态；部分原始肺泡上皮细胞分化为扁平的肺泡Ⅰ型细胞和立方形的肺泡Ⅱ型细胞；Ⅱ型细胞内出现嗜锇小体。肺泡期(16～22周)以肺泡的形成和分化为主，更多的肺泡上皮分化为扁平的肺泡Ⅰ型细胞和立方形的肺泡Ⅱ型细胞。此期，毛细血管内皮与部分肺泡上皮贴近，可将肺泡上皮细胞区分为3种：Ⅰ型细胞，呈矮柱状或椭圆形，胞质中有较明显的核糖体、扩张内质网及变性线粒体；形成了由Ⅰ型细胞一基膜一内皮细胞组成的气血屏障。Ⅱ型细胞，胞质内含丰富的嗜锇板层小体和核糖体，内质网扩张呈大小不一的泡状，多泡体出现，线粒体膨大变性，细胞游离面可见少数微绒毛。Ⅲ型细胞，为未分化细胞，呈立方形，胞体较小，胞核相对较大，呈圆或椭圆形，胞质少，呈带状．电子密度低，细胞器少。     

4种日粮条件下30～150日龄羔羊瘤胃消化代谢功能的变化

将16只30日龄体重约6～10kg装有瘤胃瘘管的罗姆尼与新疆美利奴羊杂交公羔分为4组，分别喂以不同消化能（DE，MJ／kg）和蛋白质（CP，％）水平的日粮（DE／CP：11．91／20、11．91／14、8．15／20和8．15／14），以15d为一研究时间段，每时间段的第9、10和11天在第一次喂料后的第0、1、3、6和9h采取瘤胃液样品，以研究30～150日龄期间羔羊瘤胃消化代谢的变化规律。结果表明，羔羊瘤胃液pH、氨态氮、丙酸、乙＋丙＋丁酸浓度随日龄而不断升高，在100日龄时趋于稳定，并且受日粮营养水平的影响。乙酸浓度的变化虽也受日龄和日粮营养水平的影响，但相对较小。羔羊瘤胃液体积在120日内几乎增加了15倍，并且也受日粮营养水平的影响。研究表明瘤胃消化代谢的发育是渐进性的，在100日龄时各项指标均趋于稳定，并且瘤胃液体积的增大对于瘤胃消化代谢能力的发育起着首要的作用。     

母牛一胎双犊技术的研究

选择9头母牛先行人工授精、7天后再移植冷冻胚胎以生产双犊。试验结果表明,母牛产双犊达44.44%(P<0.01)。提示:采用A(I人工授精)+ET(胚胎移植)是提高母牛双犊率的有效技术措施之一。     

核酸疫苗研究进展(下)

Sato等报道，在质粒载体上引入大肠杆菌Amp^ 抗性基因(内含有6核苷酸片段的ISS免疫增强序列)，真皮免疫小鼠后ISS增强CTL和Th1细胞的免疫反应。有人提出利用含有病毒免疫原性基因的基因大片段进行基因免疫，以弥补单基因免疫活性的不足。将协同因子如IL-2、IL-12、TCA等的表达质粒与抗原表达质粒混合注射可使Th1型细胞免疫增强，而GMcsf能明显加强体液免疫，     

Based serum metabolomics analysis reveals simultaneous interconnecting changes during chicken embryonic development

Metabolic disorder is a major health problem and is associated with a number of metabolic diseases. Due to native hyperglycaemia and resistance to exogenous insulin, chickens as a model had used in the studies of adipose tissue biology, metabolism and obesity. But no detailed information is available about the comprehensive changes of serum metabolites at different stages of chicken embryonic development. This study employed LC/MS‐QTOF to determine the changes of major functional metabolites at incubation day 14 (E14d), 19 (E19d) and hatching day 1 (H1d), and the associated pathways of differential metabolites during chicken embryonic development were analysed using Metabolite Set Enrichment Analysis method. Results showed that 39 metabolites were significantly changed from E14d to E19d and 68 metabolites were significantly altered from E19d to H1d in chicken embryos. Protein synthesis was promoted by increasing the concentrations of L‐glutamine and threonine, and gonadal development was promoted through increasing oestrone content from E14d to E19d in chicken embryos, which indicated that serum glutamine, threonine and oestrone contents may be considered as the candidate indicators for assessment of early embryonic development. 2‐oxoglutaric acid mainly contributed to enhancing the citric cycle, and it plays an important role in improving the growth of chicken embryos at the late development; the decreasing of L‐glutamine, L‐isoleucine and L‐leucine contents from E19d to H1d in chicken embryonic development implied their possible functions as the feed additive during early posthatch period of broiler chickens to satisfy the growth. These results provided insights into understand the roles of serum metabolites at different developmental stages of chicken embryos, it also provides available information for chicken as a model to study metabolic disease or human obesity.     

Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Differential gene expression of Eph‐ephrin A1 and LEPR‐LEP with high or low number of embryos in pigs during implantation

The objective of this study was to ascertain whether mRNA and protein expressions of implantation‐related genes (erythropoietin‐producing hepatocellular receptor–ligand A1, Eph‐ephrin A1 and leptin receptor–leptin, LEPR‐LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre‐pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO‐NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion–allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high‐embryo compared to the low‐embryo group, respectively (p < .05). Real‐time qPCR results showed that Eph‐ephrin A1 mRNA expression was less in the high‐embryo (p < .05) compared to the low‐embryo group. In addition, Western blotting analysis indicated that Eph‐ephrin A1 and LEP protein expression at endometrial attachment site in high‐embryo was less (p < .05) compared to low‐embryo group. It was also noted that mRNA expression of Eph‐ephrin A1 and LEPR‐LEP was greater in pregnant than non‐pregnant gilts (p < .05). Moreover, mRNA expression of Eph‐ephrin A1 (p < .05) and LEPR‐LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph‐ephrin A1, LEPR‐LEP and embryo length with the correlation coefficient 0.31–0.59. For Eph‐ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR‐LEP, the highest correlation coefficient appeared between LEPR‐LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph‐ephrin A1 and LEPR‐LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph‐ephrin A1 and LEPR‐LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph‐ephrin A1 and LEPR‐LEP might play roles in the regulation of embryo implantation in pigs.     

Effect of parental origin on early life history traits of European eel

Establishment of European eel (Anguilla anguilla) hatchery production will rely on selectively bred individuals that produce progeny with the best traits in successive generations. As such, this study used a quantitative genetic breeding design, between four females and nine males (four wild‐caught and five cultured), to investigate the effect of paternal origin (wild‐caught vs. cultured) and quantify the relative importance of parental effects, including genetic compatibility, on early life history (ELH) performance traits (i.e. fertilization success, embryonic survival at 32 hr post‐fertilization, hatch success and larval deformities at 2 days post‐hatch) of European eel. Wild‐caught males had higher (56%) spermatocrit values than cultured males (45%), while fertilization success, embryonic survival, hatch success and larval deformities were not significantly impacted by paternal origin. This demonstrates that short‐term domestication of male eels does not negatively affect offspring quality and enables the consideration of cultured male broodstock in future breeding programmes. Moreover, paternity significantly explained 9.5% of the variability in embryonic survival, providing further evidence that paternal effects need to be taken into consideration in assisted reproduction protocols. Furthermore, maternity significantly explained 54.8% of the variation for fertilization success, 61.7% for embryonic survival, 88.1% for hatching success and 62.8% for larval deformities, validating that maternity is a major factor influencing these “critical” ELH traits. At last, the parental interaction explained 12.8% of the variation for fertilization success, 8.3% for embryonic survival, 4.5% for hatch success and 20.5% for larval deformities. Thus, we conclude that eggs of one female can develop more successfully when crossed with a compatible male, highlighting the importance of mate choice for successful propagation of high‐quality offspring. Together, this knowledge will improve early offspring performance, leading to future breeding programmes for this critically endangered and economically important species.