Comparison of the Value of Pulsed-field Gel Electrophoresis,Random Amplified Polymorphic DNA and Amplified rDNA Restriction Analysis for Subtyping Taylorella equigenitalis

Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzymes, including AluI and MboI, of the 1,500 bp fragments of rDNA amplified by polymerase chain reaction did not discriminate the genomic variations among the eight strains of T equigenitalis. Thus, pulsed-field gel electrophoresis was shown to discriminate these eight organisms better than random amplified polymorphic DNA analysis, while amplified rDNA restriction analysis was found to be unsuitable for subtyping T equigenitalis.     

猪伪狂犬病病毒JSSQ2013株的分离鉴定及重要功能基因序列分析

为了解江苏省猪伪狂犬病病毒(Pseudorabies virus,PRV)野毒株的特点,本研究从2013年采自江苏省宿迁市的疑似PRV感染病料中分离纯化了一株PRV病毒,对其进行了PCR和间接免疫荧光法(IFA)鉴定,并进一步在Vero细胞上测定该分离株的病毒滴度TCID50和一步生长曲线,扩增其gB、gC、gD和gE基因进行序列比对及分子遗传进化分析,并将该分离株分别接种新西兰白兔和15日龄仔猪研究其致病性。结果显示,该病毒为一株PRV,命名为PRV JSSQ2013株,纯化后的病毒滴度为10^7.8 TCID50/ml;生长曲线测定显示在感染20h后病毒滴度即达到最高,为10^8.6 TCID50/ml。与我国近几年分离的PRV变异株序列相比,PRV JSSQ2013株的gB、gC、gD和gE基因核苷酸序列同源性分别为99.5~99.6%、99.5~99.6%、99.5~99.6%和98.7~99.7%,氨基酸序列同源性分别为98.9~99.0%、99.5~99.7%、99.0~99.2%和98.1~99.3%,均高于其与经典毒株(Ea、Fa和SC株)和欧美毒株(Becker、Kaplan、Bartha、Kolchis和NIA3)的同源性;基于gB、gC、gD和gE基因的遗传进化树分析均显示PRV JSSQ2013株与国内近几年分离的PRV变异株属同一分支。该病毒接种新西兰白兔后均出现典型的PR症状,如厌食、兴奋、啃咬或用爪挠接种部位等典型症状,且在48h内全部死亡;接种仔猪后第1天开始出现典型的PR症状,第5天全部死亡。以上结果证实,从江苏省宿迁市采集的疑似PRV感染病料中分离到一株强毒力的PRV变异株。本研究为了解江苏PRV分子流行特征、丰富我国PRV分子流行病学资料及新型疫苗的研制奠定了基础。     

一个紫云英F;重组自交系群体的农艺性状与养分吸收评价北大核心CSCD

利用以闽紫7号紫云英为母本、信阳种紫云英为父本获得的F;代群体,评价了该群体18个株系的农艺性状和养分吸收状况,为筛选优异紫云英株系做参考。结果表明,在盆栽条件下,不同株系紫云英在盛花期表现出一定的性状变异。相关性分析表明,鲜重、干重与氮、磷、钾积累量相关性高,对植株的养分吸收影响较大;株高、分枝数、真叶数是影响植株地上部鲜重、干重的重要因素。主成分分析表明,地上部鲜重、干重以及地上部氮、磷、钾积累量是反映紫云英株系性状优劣的主要指标。聚类分析结果显示,当距离系数d=25时,18份紫云英材料可分为3类,其中Ⅰ类群综合性状最优,包括m83xzh-1-4-4、m83xzh-1-4-6、m83xzh-1-4-8和m83xzh-1-4-13共4个株系。综合考虑紫云英生长性状、鲜草产量及植株养分积累量等因素,m83xzh-1-4-4的地上部磷、钾积累量和地下部氮、钾积累量最高,每盆分别高达11.00 mg,145.18 mg,22.15 mg和15.27 mg,同时该株系具有较高的鲜草产量,每盆高达44.63 g。本研究可为后续按目标性状选育紫云英新品种提供参考。     

菠萝皮发酵生产饲料蛋白优良菌种的筛选

以菠萝皮为原料,经烘干粉碎后添加一定辅料,采用10株不同的霉菌和酵母菌株进行单一菌种发酵试验,预筛选出产蛋白含量较高的3株霉菌和1株酵母菌;再将预筛选出的霉菌和酵母菌经过不同组合按一定的比例和接种量对菠萝皮渣进行混和菌种发酵。结果表明:采用绿色木霉(Trichodermaviride)和产朊假丝酵母(Candicau-tilis)按3:2的接种比例、5%的总接种量进行发酵,可使菠萝皮发酵产物的粗蛋白质含量和水洗蛋白含量显著提高,且发酵产物气味芳香,适口性好,可做动物蛋白饲料。     

Serological response of guinea pigs to oily and aqueous inactivated vaccines containing a Brazilian isolate of the Bovine Viral Diarrhea Virus (BVDV)

Bovine Viral Diarrhea Virus (BVDV) is widespread in cattle in Brazil and research shows its large antigenic variability. Available vaccines are produced with virus strains isolated in other countries and may not be effective. In this study, inactivated vaccines containing the Brazilian BVDV-Ib IBSP11 isolate were developed and tested on 6 groups of 10 guinea pigs (Cavia porcellus). Animals in groups A and C received an aqueous vaccine (aluminum hydroxide); B and D groups received an oily vaccine (Montanide ISA50); Group E positive-control animals were given an imported commercial vaccine with BVDV-Ia Singer; Group F animals were sham vaccinated (negative control). Groups A, B and E received two doses, and Groups C and D, three, every 21 days. Twelve blood samples were taken, at 21-day intervals over 231 days, and evaluated for antibody titer through virus-neutralization (VN), using a homologous strain (IBSP11), and a heterologous strain (BVDV-Ia NADL). Most animals, 42 days following the first dose, seroconverted to both strains and, after the second dose, there was a significant increase of titers in all groups. The oily formulation induced greater response after the third administration. This increase was not observed with the aqueous vaccines, regardless of the virus used in the VN. Antibody decline was more rapid in animals that received aqueous vaccines. The results showed the importance of studying the influence of endemic strains of commercial vaccines, to improve the efficacy of BVD vaccination. Use of the endemic strain in vaccine formulation presented promising results, as well as the use of guinea pigs as a laboratory model.     

不同微生物添加剂组合固态发酵对木薯渣品质的影响

本试验旨在研究布氏乳杆菌(LAB)、黑曲霉(AN)、热带假丝酵母(CT)、枯草芽孢杆菌(BS)与植物乳杆菌(LAP)组合对木薯渣品质的影响,筛选出发酵效果最优的混合菌组合。试验以木薯渣为发酵原料,对LAB、AN、CT、BS与LAP进行不同菌种的菌液以1∶1体积比组合,共设5个不同组合,组合1:LAB+AN+CT+LAP;组合2:LAB+AN+BS+LAP;组合3:LAB+CT+BS+LAP;组合4:AN+CT+BS+LAP;组合5:LAB+AN+CT+BS+LAP;各组合分别添加1%尿素或者1%尿素+0.6%红糖,空白组不添加任何添加剂,对照组Ⅰ添加1%尿素;对照组Ⅱ添加1%尿素+0.6%红糖,各组均用生理盐水调制含水量为65%左右,于聚乙烯薄膜袋中真空发酵10 d。结果表明:1)不同微生物添加剂组合发酵木薯渣对木薯渣营养成分改善效果5菌组合优于4菌组合。其中,LAB+AN+CT+BS+LAP+尿素+红糖组合发酵木薯渣效果最好,与空白组相比显著降低了发酵木薯渣p H(P0.05),提高了乙酸和丙酸含量;中性洗涤纤维(NDF)和酸性洗涤纤维(ADF)含量均为最低,显著低于空白组和对照组(P0.05);粗蛋白质(CP)含量最高,显著高于空白组和对照组(P0.05)。2)添加相应的菌种+尿素+红糖发酵木薯渣有利于产生丙酸。3)在发酵培养基中添加尿素能显著提高木薯渣CP含量(P0.05)。4)在木薯渣发酵过程中添加尿素+红糖比只添加尿素对木薯渣营养成分改善效果好。综上所述,LAB、AN、CT、BS与LAP组合添加尿素和红糖固态发酵木薯渣,可以有效改善木薯渣品质。     

复合促生菌剂发酵条件优化及其对青稞促生效果评价

为研制复合植物根际促生菌(Plant growth promoting rhizobacteria,PGPR)菌剂,提升菌体生物量,提高菌剂促生能力,本研究以3株分离自高寒草地的优良PGPR菌株组成复合菌系,通过单因素试验及响应面法Box-Be-hnken试验设计,优化复合菌系发酵培养基成分及培养条件,并通过盆栽试验评...     

草地早熟禾根际固氮菌的生物效能

利用乙炔还原法、比色法等对草地早熟禾(Poa pratensis)根际分离得到的21个固氮菌株进行固氮酶活性、溶磷能力和分泌植物生长素性能的研究测定,在固氮酶活性测定中,21个菌株还原C2H2产生的C2H4在39.9～227.5 nmol·mL-1·h-1,大于100 nmol·mL-1·h-1的有14株,占所分离固氮菌株的67%;菌株的溶磷强度在13.38～58.21 μg·mL-1,其中菌株N4 、N16及N20溶磷能力较强;菌株N2、N4、N5、N10、N14、N16、N17及N20具有较强的分泌植物生长激素的能力。     

贵州尾矿区野生豆科牧草根瘤菌及其抗性测定

为了筛选获得对复合重金属具有耐受能力且能在本地豆科植物根系内定殖共生的抗性根瘤菌,本研究从贵州省内的锰矿区、铅锌矿区采集野生豆科牧草根瘤样品,经分离纯化、菌体形态观察、16S rRNA基因序列比对分析,获得71个根瘤菌保存菌株。系统发育和区系分析结果表明,分离菌株分属7属28种,其中根瘤菌属(Rhizobium)的分布频率高达54.93%,为优势属,木兰根瘤菌(R.indigoferae)的分布频率最高(15.49%),为优势种。选择来自不同尾矿区、不同种类的根瘤菌17株,采用重金属平板法测定各供试菌株对贵州尾矿土分布较多的Pb²⁺,Zn²⁺,Cr⁶⁺,Mn²⁺,Cd²⁺五种重金属的耐受性,结果发现17个菌株对3种或3种以上的重金属都具有不同程度的耐受能力,其中HMZT039-6,HSST042-6,HSST042-1,ZHQT052-1和HXTT050-1对5种重金属离子均有耐受能力,为筛选出的最优抗性根瘤菌株。本研究结果可为本地区土壤重金属污染的修复治理提供共生微生物资源。     

2008年江苏地区基因Ⅶd亚型新城疫病毒遗传变异分析

为了研究基因Ⅶ d亚型新城疫病毒(Newcastle disease virus,NDV)的分子流行病学及遗传变异规律,作者对2008年分离自江苏地区16株具有一定代表性NDV分离株的F和HN基因片段进行了扩增.根据F基因和HN基因序列绘制的2个遗传进化树基本一致,表明分离株中不存在囊膜糖蛋白基因发生重组的NDV;同时,HN蛋白线性表位发生E347K突变的变异株的分离率在我国呈上升趋势.基因Ⅶd亚型变异株的出现应引起相关领域从业者们的高度重视.