山羊皮肤新microRNA的挖掘

已有证据表明,microRNA在动物皮毛发育和形态发生过程中发挥重要的作用.在山羊皮肤microRNA的研究中,数据库不完善仍是限制其功能研究的主要原因.本研究以山羊基因组为参考,对高通量测序中未注释的占总reads 5.40%的数据,使用Mireap软件预测的方法进行数据挖掘,并对新的microRNA的长度、表达量、基因组定位、靶基因预测及KEGG pathway进行分析.研究预测到28个新的microRNA,其长度分布与已知山羊、牛等动物的分布一致.对表达量统计表明,表达量在100以上的有4个,最高的是Novel-15,达到555;表达量在10~100的有19个;低于10的有5个.基因组定位研究表明,新microRNA在山羊染色体上分布均匀,在5、6、7、19和X染色体上分别有两个microRNA被定位.预测和分析得到6 519个靶基因.KEGG分析结果表明,有3 552个靶基因定位到51条通路中,占前3位的通路是Metabolic pathways、Pathways in cancer、MAPK signaling pathway.本研究发掘了更多山羊的microRNA,为今后山羊皮肤发育调控的研究奠定了基础.     

醋酸铅对母鼠生殖与胚胎发育影响

将420只体质量为(15.32±2.01)g的雌性昆明系小鼠随机分为5个处理组和1个对照组,每组设7个重复,每个重复10只。各处理组分别腹腔按体质量注射10、15、20、25、30mg/kg的醋酸铅溶液,对照组注射等体积的灭菌生理盐水,每隔2d注射并称重1次,共注射10次,期间记录小鼠体质量及临床表现。当小鼠体质量达到25g以上时,分批对各试验组和对照组进行超排处理,腹腔注射10IU马绒毛膜促性腺激素(PMSG),47h后注射10IU人绒毛膜促性腺激素(hCG),并与公鼠合笼。合笼后87~96h内颈椎脱臼处死小鼠,观察卵巢、子宫形态,并统计胚胎数,同时制作卵巢、子宫石蜡切片,观察其病理组织学变化,研究醋酸铅对雌性小鼠卵巢、子宫组织结构及早期胚胎发育的影响。结果显示:(1)当醋酸铅染毒剂量≥20mg/kg时,可明显抑制小鼠体质量的增长,随着染毒剂量的增加,作用时间的延长,小鼠体质量增加明显趋缓,与对照组相比,差异显著或极显著(P0.05,P0.01);(2)染铅组母鼠早期胚胎发育受到显著影响,主要表现为回收胚胎总数以及受精卵发育到桑椹胚和囊胚的总数均显著低于对照组(P0.05),而各染铅组退化胚、延迟胚数和未受精卵总数显著高于对照组(P0.05);(3)染铅组卵巢中的初级卵泡、次级卵泡、成熟卵泡数量明显低于对照组,而原始卵泡、闭锁卵泡数量明显高于对照组。(4)染铅组小鼠卵巢和子宫形态发生明显畸形,当醋酸铅染毒剂量≥20mg/kg时,与对照组相比,差异显著或极显著(P0.05,P0.01)。且上述变化均呈明显的剂量一时间效应。研究结果表明,当醋酸铅暴露剂量≥20mg/kg时,可对小鼠生长发育具有明显抑制作用,同时使母鼠生殖器官卵巢和子宫的结构造成严重损害,并影响其生殖功能与早期胚胎发育。     

饲料中添加天蚕素抗菌肽对南美白对虾免疫功能、抗病力的影响

本研究以南美白对虾为研究对象,探究饲料中天蚕素抗菌肽的合适添加量。在饲料中依次添加不同浓度水平（0%、1%、2%、3%、4%和5%）的天蚕素抗菌肽,配制为6种配合饲料,用以投喂南美白对虾,饲养周期8周。结果显示：与对照组相比,饲料中添加天蚕抗菌肽对南美白对虾增重率、蛋白质沉积率和特定生长率有显著影响（P＜0.05）,高水平天蚕素抗菌肽会抑制南美白对虾的增重率和特定生长率,南美白对虾的饵料系数显著下降,但对南美白对虾存活率和摄食率无显著影响（P＞0.05）|随着饲料中天蚕素抗菌肽的添加水平不断增加,3%组南美白对虾的ACP、POD和SOD含量达到最大水平,与对照组有明显差异（P＜0.05）|随着饲料中天蚕素抗菌肽的添加水平的不断上升,3%组南美白对虾的淀粉酶、脂肪酶和胃蛋白酶含量最高,与对照组存在明显差异（P＜0.05）。结果表明,饲料中添加天蚕素抗菌肽显著提高南美白对虾的增重率和特定生长率,提高南美白对虾的生长发育速度,从而增强机体免疫力,提高南美白对虾的抗病能力。 [关键词]天蚕抗菌肽|南美白对虾|抗病力|免疫功能     

Influence of dietary lysine level on whole-body protein turnover, plasma IGF-I, GH and insulin concentration in growing pigs

Four growing pigs (initial liveweight 25.9 ± 0.54 kg, final liveweight 43.0 ± 1.06 kg) were used to study the effect of dietary lysine level on nutrient digestibility, whole-body protein turnover, plasma insulin-like growth factor-I (IGF-I), growth hormone (GH), insulin, glucose, and urea nitrogen (PUN). Four diets, containing 7.0 g (L1), 9.5 g (L2), 12.0 g (L3) and 14.5 g (L4) lysine per kg diet respectively, were formulated as experimental treatments. The animals and diets were allocated in a 4 × 4 Latin square design. Nitrogen (N) metabolism and whole-body protein turnover were measured by classical method and single-dose ¹⁵N end-product method, respectively. The blood samples were taken at the end of each experimental period. Results showed that N retention (NR) and N biological value (NBV) were significantly increased from L1 to L4 (P < 0.05). However, differences in NR and NBV between L2, L3 and L4 were not significant (P > 0.05). There was no significant difference on dry matter (DM) digestibility, organic matter (OM) digestibility and N digestibility between different treatments (P > 0.05). Whole-body protein synthesis, protein degradation and protein accretion increased markedly from L1 to L2 (P < 0.05), but did not increase further from L2 to L4. Whole-body protein accretion (y, g/kg W^0.75/d) increased with dietary lysine (x, g/kg) in a quadratic manner: y = − 0.09x² + 2.12x − 5.14 (r² = 0.96, n = 4, P < 0.05).The results also showed that differences in plasma IGF-I, GH, glucose and PUN concentration between different treatments were not significant (P > 0.05). Plasma insulin concentration (y, μIU/ml) was increased with dietary lysine (x, g/kg) in a quadratic manner: y = 0.23x² − 4.10x + 32.25 (r² = 0.99, n = 4, P < 0.05), but it was not found that plasma insulin concentration was related to NR. A significant correlation was found between NR (y, g/d) and plasma IGF-I (x, ng/ml): y = − 3.1 × 10^− 3x² + 1.31x − 122.28 (r² = 0.99, n = 4, P < 0.05).It was concluded that dietary lysine level had a significant influence on NR and whole-body protein turnover but not on plasma IGF-I and GH concentration. Plasma IGF-I may be an important factor controlling N metabolism of growing pigs. Further research was needed to study the mechanism.     

Generation of a genotype VII Newcastle disease virus vaccine candidate with high yield in embryonated chicken eggs

To generate a genotype VII Newcastle disease virus (NDV) vaccine with high yield in embryonated chicken eggs, we selected genotype VII NDV strain JS5/05, which possesses a high virus titer in embryos as the parental virus. Using reverse genetics, we generated a genetically tagged derivative (NDV/AI4) of JS5/05 by changing the amino acid sequence of the cleavage site of the F0 protein. Pathogenicity tests showed that NDV/AI4 was completely avirulent. NDV/AI4 was genetically stable and replicated efficiently during 10 consecutive passages in embryos. More importantly, serologic assays showed that oil-emulsion NDV/AI4 induced higher hemagglutination inhibition (HI) titers against the prevalent virus than oil-emulsion LaSota vaccine in chickens and geese. Moreover, NDV/AI4-induced HI titers rose faster than those elicited by LaSota in chickens. Both NDV/AI4 and LaSota provided protection against clinical disease and mortality after the challenge with the genotype VII NDV strain JS3/05. However, NDV/AI4 significantly reduced virus shedding from the vaccinated birds compared to LaSota. Taken together, these results suggest that NDV/AI4 can provide better protection than LaSota and is a promising vaccine candidate against genotype VII NDV.     

Rapid detection of SNPs in candidate genes regulating the growth of orange‐spotted grouper,Epinephelus coioides (Hamilton, 1822), using semiconductor sequencing

Orange‐spotted grouper (Epinephelus coioides) is one of the most important marine food species in Southeast Asia and China and has been cultured for decades. In this study, we fully utilized the limited capacity of semiconductor sequencing, the high efficiency of long‐range PCR for target enrichment and a non‐indexed pooling strategy to screen single‐nucleotide polymorphisms (SNPs) in a breeding population of orange‐spotted grouper. Forty‐one genomic DNA fragments, with a total length of approximately 180 kb, including 22 candidate genes that control growth, and from a DNA pool of 20 heaviest and 22 lightest individuals of the sampled population were successfully sequenced using an Ion Torrent Personal Genome Machine. 3 503 466 clear reads were produced with a length of 192 ± 56 bp, 86.8% of which were mapped to the reference with an average coverage depth of 2567‐fold and physical coverage of 98.8%. Finally, 1623 high‐quality SNPs were adopted. Compared with Sanger sequencing of three random common regions, the sensitivity and specificity of our approach were 39.4% and 100.0% respectively. A mutation located at the third position of the previously labelled start codon of growth hormone receptor type 1 invalidated the start codon. Furthermore, comparison of the frequencies of genotypes and alleles of this site between the two extreme groups, prediction of signal peptide and identification of conservative mRNA sequences suggested that the functional start codon is likely located at the position of another downstream in‐frame ATG in the mutant. These detected SNP markers will provide important tools for the selective breeding of orange‐spotted grouper.     

Research Progress of Fatty Acid Regulation of microRNAs in Dairy Cow Mammary Glands

Lipids are a kind of substance which can store and provide energy for animals. A large number of studies have confirmed that the change of lipids in diets can alter the expression of microRNAs (miRNAs) and thus affect the biological activities of the bodies. This review mainly summarizes the regulation of fatty acids on the expression of miRNAs in mammary gland of dairy cows, from the aspects of bovine mammary miRNAs expression characteristics, miRNAs that regulate milk fat synthesis, and the changed bovine mammary miRNAs in response to exogenous addition of fatty acids. All these works may be beneficial for the establishment of the connection between nutrients and mammary function, and provide information for the related research in the future.     

应用计算机和气象资料预测粗纹食道口线虫（Oesophagostomum asperum）数量季节消长的研究

根据土源性寄生线虫体外发育期间受温度、湿度、季节和宿主动物影响的原理，采用电子计算机对上述四因子和食道口线虫从卵发育到侵袭性幼虫速度、数量进行回归分析，制定出模拟模型，设计出了在牧地上该种线虫从卵发育到侵袭性幼虫数量预测预报公式，用该公式和气象资料计算出的侵袭性幼虫季节消长曲线，与牧地上宿主羊体内成虫数量消长曲线相一致，初步显示可以采用气象资料和计算机预测牧地上粗纹食道口线虫侵袭性幼虫量的季节动态，从而为防治措施的制定提供依据。     

副猪嗜血杆菌S-核糖基高半胱氨酸酶基因序列分析与推导蛋白三维结构的分子模拟

利用PCR方法获得副猪嗜血杆菌S-核糖基高半胱氨酸酶(luxS)基因全长DNA,将PCR纯化产物与pMD18-T载体连接并转化E.coil DH5α菌株,重组阳性质粒测序并采用生物信息学软件对所推导的氨基酸序列进行三维结构分析。结果表明,该基因全长510bp,并与GenBank中登录的其他5株菌株luxS基因完整参考序列进行比较,同源性均在70%以上。用ExPAsy软件包预测了推导蛋白的特性,运用Swiss-PDB viewer软件的SWISS-Model处理器,并利用同源建模的思想建立HPS-luxS的三维结构。拉马钱德兰图证明,构建的luxS蛋白的空间结构是合理的。     

牛组织蛋白D基因多态性与肉质、胴体性状的关联分析

以51头西门塔尔和鲁西黄牛的杂交后代作为试验动物,扩增组织蛋白酶D(cathepsin D,CTSD)基因部分外显子,通过DNA测序方法检测到该基因第5、6、8外显子共4个SNPs(T581C、T599C、A861T、A1128G)的遗传变异。SAS软件采用最小二乘法拟合线性模型,将几个位点的不同基因型与牛肉质、胴体性状进行关联分析。结果表明,位于CTSD基因外显子5的T581C突变与大理石纹性状呈显著相关(P<0.05),BB基因型显著优于AA和AB基因型,这为T581C作为肉牛大理石纹的分子标记提供了一定的理论依据。