牛轮状病毒TaqMan实时荧光RT-PCR 快速检测方法的建立

本研究按照牛轮状病毒(BRV)结构蛋白VP6基因序列,设计合成引物和探针,经各反应条件的优化,建立了BRV TaqMan实时荧光定量RT-PCR技术。对BRV进行了特异性、敏感性和重复性试验。结果表明,TaqMan实时荧光RT-PCR最低可检测到100个拷贝病毒RNA;与牛病毒性腹泻病毒(BVD)、猪瘟病毒(CSFV)、牛结核杆菌(MB)和牛传染性鼻气管炎病毒(IBRV)不发生交叉反应;所制作的标准曲线在10²~10⁹拷贝/μL浓度范围内有极好的线性关系且线性范围宽,相关系数为0.997;与常规的RT-PCR相比,该方法具有快速、特异、敏感、重复性好、可同时检测大量样品等优点。可对样品中微量BRV进行准确检测,对BRV的诊断有重要意义。     

基于叶绿体trnL-F序列对广义披碱草属物种的系统进化研究

广义披碱草属包括了披碱草属、鹅观草属、猬草属、裂颖草属和仲彬草属。为深入研究这5个小麦族多年生属的系统地位、母本来源及St染色体组的来源和分化等问题,本研究根据叶绿体DNA trnL-F序列构建了系统树和网状结构图。结果表明,1)广义披碱草属物种的母本不完全来源于拟鹅观草属的St染色体组,其中黑药仲彬草和梭罗仲彬草的母本染色体组为P染色体组,Hystrix duthiei及长芒猬草的母本染色体组为Ns染色体组;2)系统树中反映了P、W和St染色体组之间具有较近的亲缘关系,它们与H和Ns染色体组的关系较远;3)仲彬草属中黑药仲彬草和梭罗仲彬草与冰草属的二倍体物种聚类在一起,表明黑药仲彬草和梭罗仲彬草与冰草属的关系较近;猬草属中猬草聚在St支中,而H. duthiei和长芒猬草包括在Ns支中,表明它们具有不同的染色体组组成;4)5个不同地理分布的具有St染色体组的拟鹅观草属二倍体物种没有形成一个单系组,表明St染色体组在二倍体物种中存在一定程度的分化;在异源多倍体物种中,其St染色体组的质体序列构成了不同的分支,表明St染色体组在多倍体物种中也具有分化;5)亚洲分布的拟鹅观草属二倍体物种可能参与了大部分欧亚物种的起源,北美分布物种的St染色体组可能来源于北美的拟鹅观草属物种;有些物种在多倍化起源过程中可能发生了重复杂交过程。     

酸奶及乳饮料中大肠杆菌变化趋势研究

[目的]为了探讨实际生产过程中酸奶和乳饮料感染大肠杆菌（E.coli）后的变化情况及影响因素。[方法]将大肠杆菌标准菌株接种至酸奶、乳饮料及培养基中,设置不同的储藏温度、pH,定期测定储藏过程中大肠杆菌含量变化情况。[结果]随着储藏时间延长,不同储藏温度（4℃和25℃）,不同pH(4.2和3.6)的酸奶和乳饮料中大肠杆菌含量均减少,相对于储藏于4℃的产品,储藏于25℃的产品中菌含量降低更快;当产品pH为3.6时,菌含量减少速率高于pH为4.2的产品。将大肠杆菌置于不同pH的培养基中培养,结果表明,当pH<3.8时,大肠杆菌的活性将会受到抑制,证明了pH对大肠杆菌的影响。[结论]本研究为实际生产加工过程中产品冷藏及脱冷条件下大肠杆菌的检验检测和产品质量控制提供理论依据和理论指导。     

我国部分地区牛支原体肺炎和关节炎的病原体诊断

重庆等4省市从外省引进的肉牛群发生以严重肺炎和关节炎为主要表现的疾病。该病的病变主要集中于肺脏,其组织学变化主要是间质增生,纤维素渗出,干酪样坏死,淋巴细胞、嗜中性粒细胞和单核细胞浸润。PCR检测病牛肺脏显示牛支原体阳性,而丝状支原体丝状亚种、牛分支杆菌和多杀性巴氏杆菌为阴性。从肺等组织中分离到牛支原体以及大肠杆菌、沙门氏菌、链球菌、葡萄球菌和烟曲霉,没有分离到巴氏杆菌。结果显示本病是以牛支原体感染为主引起的牛支原体肺炎和关节炎,长途运输等应激因素是该病突发的重要诱因,其他细菌和/或真菌继发感染加重了病情。     

Bioinformatics Analysis of Bone Morphogenetic Protein 4 (BMP4) of Sheep

The assay was aimed to explore the biological characteristics of bone morphogenetic protein 4 (BMP4) of sheep,NCBI,DNAMAN DNAStar,TMHMM Server v.2.0,PsortⅡ,SignalP various bioinformatical softwares were used to speculate the physical and chemical properties,hydrophobic property,phosphorylation site,conservative structure domain,protein secondary structure of BMP4 protein.Also,the three-dimensional structure was forecasted with the SWISS-MODEL Workspace software.The results indicated that the BMP4 of sheep had high homologies with the BMP4 of various species.The encoded protein was a hydrophilic protein which was unstable.There was no transmembrane regions and it was likely to be located in the nucleus.What was more,there was signal peptide and eighteen phosphorylation sites.Through the forecast of functional domains,the protein had two functional domains,including the transforming growth factor-beta (TGF-beta) superfamily and TGF-beta propeptide superfamily.The result was consistent with the function of BMP4 gene family,it also demonstrated that BMP4 was a growth factor and it had the function of signal transduction.The amino acid homology between the predicted 3D structure of protein and template 3bmp.1.A was 88.29%.The bioinformatics analysis of BMP4 gene could provide reference for the further study in practice.     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

饲料添加剂L-赖氨酸硫酸盐中L-赖氨酸含量近红外速测方法研究

为了探讨利用近红外漫反射光谱技术(NIDRS)快速定量分析饲料添加剂L-赖氨酸硫酸盐中L-赖氨酸含量的可行性,本试验在全国范围内收集了具有代表性的L-赖氨酸硫酸盐添加剂76个,采用国家标准方法对样品中的L-赖氨酸含量进行化学赋值;用光栅型近红外光谱仪扫描L-赖氨酸硫酸盐样品,获取了不同物理状态下样品的近红外光谱图。依据L-赖氨酸含量将样品分为定标集和验证集,运用适当的光谱预处理方法,采用竞争性自适应重加权(CARS)算法结合偏最小二乘法(PLS)建立了L-赖氨酸硫酸盐的近红外定标分析模型,并将该模型与全波长模型进行了比较。结果表明:用烘干、60目粉碎后的样品结合CARS算法建立的定标模型最优,定标集校正决定系数(R2C)为0.954,校正集标准偏差(SEC)为0.510,交互验证标准偏差(SECV)为0.659;验证集预测决定系数(R2P)为0.952,预测标准偏差(SEP)为0.554,相对分析误差(RPD)值为3.83。由此可见,NIDRS定量分析L-赖氨酸硫酸盐具有一定可行性,对于丰富我国氨基酸盐及其他氨基酸制品的快速检测方法具有实际的应用意义。     

草地生态系统碳循环研究进展

随着全球气候变化和陆地生态系统碳循环研究的发展,碳循环正成为草地生态系统研究的热点.本文在介绍其研究发展过程、主体内容和方向的同时,对目前的研究动态和研究方法进行系统的概述,重点分析了目前国内外草地生态系统碳循环过程研究、机制研究、研究方法及模型的发展动态,并对今后草地生态系统碳循环和相关研究提出了建议.     

青海牦牛肉食用品质及安全性的分析研究

对年龄2周岁左右的12头青海牦牛肉的食用品质及安全性指标进行了系统研究。结果表明：牦牛肉食用品质较好，肉质安全，无污染。具有肌红蛋白含量高、亲水率高、多汁性好，香味浓郁，肉质细嫩，口感适宜等特性。肉中肌红蛋白含量极显著高于黄牛（P＜0．01），肉中重金属元素和有毒有害物质限量均符合GB18406．3和NY5044的要求，是生产优质牛肉制品的原料，也是一种具有较高生物学价值的安全的肉食品，具有很大的开发价值。     

日粮中复合添加剂水平与猪肉中胆固醇水平关系的研究

近年来有许多研究表明,胆固醇与人类的心血管疾病有着直接的关系。人体内的胆固醇75%来源于自身合成,25%来自于动物性食品(肉、乳、蛋)。猪肉作为人们餐桌上常见的动物性食品,是人们外源性胆固醇的一个重要来源,因此如何降低猪肉中胆固