大肠杆菌K_(88ac)与LT(A~-B~ )抗原基因质粒在猪霍乱沙门氏菌弱毒株中的表达

应用细菌质粒转化技术,将大肠杆菌K_(?)与LT(A~-B~ )抗原基因重组质粒转入猪霍乱沙门氏菌弱毒菌苗株中.对获得的其中8个转化子进行鉴定的结果表明,转化的细菌仍保持沙门氏菌的形态、生化及抗原特性,同时可稳定地表达K(?)和LT-B两种抗原.用微量间接血凝试验、抗甘露糖豚鼠红细胞凝集试验(MRHA)、ELISA等对转化菌表达的K(?)抗原进行了测定,用间接免疫溶血试验对其表达的LT-B抗原进行了测定.结果,这两种抗原在转化的细菌中均可高效表达.电镜下观察,转化的细菌在其表面形成菌毛样结构.这种转化细菌表现出猪霍乱沙门氏菌与产肠毒素性大肠杆菌的两种抗原特性,为这种双价工程菌苗的研制提供了有价值的候选菌株.本研究结果还表明,猪霍乱沙门氏菌弱毒菌苗株可作为基因转化的有效受体菌.     

Sarcocystis hardangeri and Sarcocystis rangi n. sp. from the domestic reindeer (Rangifer tarandus) in northern Norway

Fresh preparations of microisolated sarcocysts from striated muscle of several domestic reindeer from northern Norway were examined by light microscopy. In cardiac muscle, cysts of S. grueneri were found. In skeletal muscle, cysts of S. rangiferi, S. tarandi and S. tarandivulpes were found in all samples examined. In the abdominal muscles of some reindeer, one or two other types of cysts were found.Cysts of one type were macroscopic in size, and ovoid to cylindrical in shape. The cysts were surrounded by a 8–12 µm thick layer of fibrous material, and measured 1682×910 µm. The cysts had relatively few and irregularly distributed, 20–35 µm long, and 3–5 µm wide, linguiform cyst wall protrusions, which could only be seen after removal of the fibrous layer. These cysts were classified as cysts of S. hardangeri, a species previously described from wild reindeer in southern Norway.Cysts of the other type were long and slender, measuring 5460–12700 (8994 ± 2575) × 95–280 (180 ± 50) µm. The cysts had numerous very fine, flexible, hair-like cyst wall protrusions, which were 8–10 [xm long and less than 0.5 µm thick. These cysts are considered to belong to a new Sarcocystis species of reindeer, for which the name Sarcocystis rangi n, sp. is proposed. The reindeer is recorded as the intermediate host for 6 different species of Sarcocystis.     

传染性支气管炎病毒DS10株S基因分析及其杆状病毒表达产物免疫原性鉴定

对肾型鸡传染性支气管炎病毒分离株Ck/Jiangsu/DS10/2008株(DS10株)S基因进行扩增、克隆和序列测定,并与GenBank中的其他25株参考株构建进化树进行比对分析,研究其遗传进化关系。另将S基因克隆至杆状病毒表达系统pFastBac 1载体并转染Sf9昆虫细胞进行表达,经间接免疫荧光鉴定其表达,用琼扩试验验证表达产物的反应性。S基因序列分析表明,S基因高变区域主要集中在157～230位、349～433位、787～886位之间,裂解位点序列为SHRRRRμSI。遗传进化分析表明,DS10与Vic株处于同一分支,而与H120等常用疫苗株相距较远。经间接免疫荧光检测表明,S蛋白在Sf9细胞中得到表达,琼扩试验验证表达产物具有良好的反应性,表明所表达的S蛋白具有良好的生物学活性。本研究表达的S蛋白可为传染性支气管炎诊断抗原的研制、新型疫苗的开发等提供基础材料。     

Production and characterization of monoclonal antibodies to (poly100)S1 protein of avian infectious bronchitis virus

Fragments within S1 genes ((poly100)S1) of infectious bronchitis virus (IBV) strains ZJ971, M41 and SC021202 (SC) were subcloned into a prokaryotic expression vector and expressed in Escherichia coli. Monoclonal antibodies (mAbs) against the recombinant (poly100)S1 proteins were produced, characterized and used to analyse epitopes on the S1 subunit of IBV. Nine mAbs raising from the three (poly100)S1 proteins recognized five different epitopes of the S1 subunit, designated as S1-A, B, C, D and E. Epitopes S1-C and S1-D are common for the three IBV strains, while S1-A and S1-B exist on ZJ971 and M41 strains, and S1-E was a strain-specific epitope for SC strain. Immunocytochemistry indicated that all the mAbs to the (poly100)S1 proteins can react with the homologous S1 glycoprotein expressed in Vero cells. Moreover neutralization test demonstrated that only mAbs 6E2, 4F9 and 6G4 had neutralization activity for the homologous IBV. These mAbs to (poly100)S1 protein were potential candidates for detecting and distinguishing IBV strains, and also used to examine antigenic variation of the S1 protein.     

7种抗菌药物对金黄色葡萄球菌及大肠杆菌体外抗生素后效应的研究

采用稀释法去除抗生素，用菌落计数测定细菌生长曲线的方法测定环丙沙星、洛美沙星、诺氟沙星、恩诺沙星、庆大霉素、青霉素及氨苄西林对金葡球菌及大肠杆菌的抗生素后效应(PAE)。结果：几种药物在各浓度组对金黄色葡萄球菌均呈现明显的PAE，氟喹诺酮类药物及庆大霉素对大肠杆菌产生较长的PAE，而β-内酰胺类药物作用后的PAE则很短。PAE的存在提示：在兽医临床设计给药方案时，适当延长给药间隔时间，减少给药次数，仍能维持抗菌效果。     

苦参碱的提取纯化工艺研究

为了优化苦参碱的提取工艺,以浸膏收率、苦参碱含量为评价指标,选择用水量、提取时间及提取次数为考察因素,利用L9（3^4）正交试验法优选出水煎煮法提取苦参碱的最佳工艺条件为用8倍量水提取3次,每次提取2h。采用乙醇沉淀及三氯甲烷萃取相结合的方法对苦参粗提物进行纯化,浓缩药液浓度为1．0g生药／mL时进行醇沉,醇沉后调节药液的pH值至9～11用三氯甲烷萃取3次。优选出的提取工艺稳定、可行,可用于苦参碱的工业化提取。     

无籽刺梨染色体数目观察

以无籽刺梨组培苗根尖为材料,探讨其染色体制片技术。结果表明,无籽刺梨根尖在常温下用0.003mol/L8-羟基喹啉溶液预处理6-8h或0.004mol/L8-羟基喹啉溶液预处理4-6h后,在预冷的卡诺固定液（冰醋酸：无水乙醇=3:1中）0℃下处理15min,经95%乙醇:15% Hcl=1:1的解离液处理5min或无水乙醇:36-38% Hcl=1:1处理6min,然后用卡宝品红染色15或20min,其镜检效果较佳;无籽刺梨的染色体数目为2n=2x=14。无籽刺梨染色体制片技术研究及其染色体数目的确定对其遗传改良工作有着重要的意义。     

长春市市售猪肉鸡肉中金黄色葡萄菌和沙门氏菌分离及耐药性分析

对肉品中金黄色葡萄球菌及沙门氏菌进行耐药分析,以期为其风险评估提供依据。采集吉林省长春市市售猪肉、鸡肉样品,按照国标方法分离金黄色葡萄球菌、沙门氏菌,利用K-B法药敏试验检测金黄色葡萄球菌和沙门氏菌对头孢噻肟钠、左旋氧氟沙星、庆大霉素、四环素4种抗生素的耐药性。结果显示：共分离出金黄色葡萄球菌46株,检出率为46％;沙门氏菌48株,检出率为16．84％。金黄色葡萄球菌对4种抗生素的耐药率在0～56．52％之间,共产生6种耐药谱。沙门氏菌对4种抗生素的耐药率在16．67％～87．5％之间,共产生9种耐药谱。长春市肉品中金黄色葡萄球菌、沙门氏菌耐药现象普遍。     

单诺沙星对大肠杆菌和金葡球菌的体外抗茵后效应

采用菌落计数法测定了单诺沙星在三种不同浓度下对大肠杆菌和金黄色葡萄球菌 (金葡球菌 )的体外抗菌后效应 (PAE)。当药物浓度为 0 .5MIC、2MIC、4MIC时 ,对大肠杆菌的PAE值为 (0 .6 4 6 4± 0 .0 2 94 )h、(1.2 0 77± 0 .0 2 84 )h、(1.6 5 2 9± 0 .0 4 96 )h ;对金葡球菌的PAE值为 (0 .5 6 6 0± 0 .0 0 75 )h、(1.174 6± 0 .0 0 5 7)h、(1.4 913± 0 .0 2 5 7)h。这表明单诺沙星对大肠杆菌、金葡球菌均有不同程度的较强的PAE ,且PAE值与药物浓度呈正相关     

停乳链球菌内蒙分离株抗原基因MIG的克隆

停乳链球菌是引起奶牛乳房炎的主要病原菌,根据GenBank中登陆的停乳链球菌表面蛋白MIG基因序列,设计一对引物,采用PCR的方法从临床分离的停乳链球菌内蒙分离株基因组DNA中扩增出MIG基因,得到一条1900bp的片段。将其连入PMD-19T载体中,经酶切,PCR及序列测定法进行鉴定。经DNA测序分析,证实与GenBank中停乳链球菌MIG基因(AF354651)序列同源性为95%,具有高度保守性。     

中美牧草种子生产比较

牧草种子在草原植被恢复和草牧业生产中发挥着重要作用。中美都是国土辽阔的大国,本文通过对比近十年内中美2国牧草种子生产的相关数据,分析中美草种业在牧草种子生产规模、专业化生产的地域性、新品种选育、产业化与机械化以及生产技术研究上的差异。结果表明：尽管我国在牧草种子专业化生产的地域性方面取得了长足进步,但是种植面积和单产仍然低于美国,每年仍需进口大量牧草种子来满足生态建设和草产业发展的迫切需求;与美国相比,我国牧草种子生产在育种、产业化、机械化和专业化等方面存在巨大的潜力,而且我国牧草种子产业亟需培育大型龙头企业。基于中美在牧草种子生产体系上的差异,本文比较分析了中美两国在牧草种子生产方面的差异和我国的发展潜力,并提出了当前我国牧草种子产业发展需要集中努力的方向和建议,希望能为推动我国草种业快速健康发展提供参考。     

Economic Impacts of the U.S. Gulf of Mexico Recreational For-Hire Fishing Industry

IMPLAN Version 3.0 was used with a 2008 IMPLAN data package to estimate the economic impacts of the recreational for-hire (RFH) fishing industry in the U.S. Gulf of Mexico (GOM). Initial, direct, indirect, and induced impacts were estimated. Impacts were estimated for head, charter, and guide boat operations for Texas, Louisiana, Mississippi, and Alabama (combined), and West Florida as individual state models, while introducing multiregional spillover effects that account for regional linkages between states. These updated data are necessary to meet national standards set forth by the Magnuson-Stevens Act, such that social and economic ramifications of proposed federal fishing regulations in the GOM can be made part of the overall decision- and policymaking process.     

中美猪肉产业互补性对双边猪肉贸易的影响分析

中美两国猪肉产业资源禀赋和生产率存在显著差异,双边猪肉贸易不平衡,产业互补性和贸易潜力有待提升。为研究中美两国猪肉产业互补性问题,本文以双边猪肉产业互补性指数为核心解释变量,对两国猪肉贸易引力模型进行了估计。结果显示:中美两国猪肉产业互补性指数对双边猪肉贸易有着正的影响;中美两国猪肉贸易潜力巨大,双边猪肉贸易具有很大的提升空间。在中美贸易纠纷逐步解决的背景下两国应加强猪肉产业合作,提高双边猪肉贸易互补水平。     

牛体中人葡萄球菌的分离与鉴定

云南省兰坪县境内期井自然村有153头黄牛发病，其主要症状为舌咽部长有类似鸡眼状麻籽样颗粒：剖检病变为咽喉充血，肺充血，脾脏稍肿一对一般的抗菌药物如青霉素、链霉素均无良好疗效经对送检病料的分离与鉴定．分离出一株人葡萄球菌。     

房山紫堇叶绿体基因组特征及其系统进化关系

房山紫堇(Corydalis fangshanensis W. T. Wang ex S. Y. He)是罂粟科紫堇属植物,根具有清热解毒的功效,也可用于防风固沙,保持水土。为探究房山紫堇的叶绿体基因组特征及其系统进化关系,利用高通量测序技术对房山紫堇叶绿体基因组进行测序和功能注释,并对其进行分析,利用MEGA7构建系统发育树。结果显示：房山紫堇的叶绿体基因组为191 128 bp, GC含量为40.2%;共注释127个基因,包括84个蛋白质编码基因(66.1%)、35个tRNA基因(27.6%)以及8个rRNA基因(6.3%);叶绿体基因组的密码子偏好性较弱;叶绿体基因组的GC3 s含量为38.5%,GC含量也小于50%,密码子偏向使用A和T这两种碱基;多重比较分析结果显示,紫堇属植物的编码区较为保守;系统发育分析表明,房山紫堇与紫堇的亲缘关系最近。研究房山紫堇叶绿体基因组及其进化关系有助于探索紫堇属植物的分类与进化,并可促进房山紫堇的开发和利用。     

家蚕核糖体18S RNA基因的序列分析及分子系统学研究

为研究家蚕18S DNA基因的特点及分子进化,以家蚕丝腺为材料提取家蚕基因组DNA,通过PCR扩增、测序鳞翅目家蚕(Bombyx mori)核糖体小亚基18S RNA基因(18S rDNA)的全序列,将该序列与等翅目、直翅目、衤责翅目、鞘翅目、膜翅目、双翅目、捻翅目、弹尾目、蜉蝣目各一种昆虫的18S rDNA进行了比较。用DNAstav软件分析并进行序列比对,结果表明,昆虫18S rDNA有4段序列较为保守,以18S rDNA比对的第2保守区段构建的分子系统发育树表明鳞翅目和双翅目、膜翅目、鞘翅目进化关系最近,等翅目、直翅目、衤责翅目进化上较为接近,捻翅目昆虫与弹尾目昆虫的亲缘关系较为接近,捻翅目在分类上应是一种古老的昆虫。     

幼狐犬瘟热与鼠伤寒混合感染的诊治报告

犬瘟热病毒与鼠伤寒沙门氏杆菌是危害狐狸的重要传染病，二者混合感染常造成大批狐狸发病死亡。本报告报道一自然发病病例，通过临床检查，实验室诊断等手段确诊为幼狐的犬瘟热与鼠伤寒混合感染，并果断采用中西结合的综合治疗措施，控制了病情，取得了满意效果。     

驯鹿18S rRNA基因的克隆测序

对驯鹿 (Rangifertarandus)的 1 8SrRNA基因片段进行了克隆测序 ,序列长度为 1 1 47bp ,并且已被发送到Genebank上 ,其登录号是AY1 45 5 2 7。     

传染性支气管炎病毒纤突蛋白S1基因的T/A载体克隆策略

参考Genbank收录的IBV纤突蛋白 (S1)基因序列 ,自行设计合成一对引物 ,对传染性支气管炎病毒 (IBV)江苏省地方分离毒株 (JS/95/0 3)RNA进行RT PCR扩增 ,产物经琼脂糖凝胶电泳分析 ,呈现一条 1716bp的条带 ,将其克隆入T/A质粒pMD18 T载体中 ,转化大肠杆菌JM10 9,挑选阳性克隆 ,用质粒少量提取法提取重组质粒 ,用EcoRⅠ和HindⅢ双酶切对重组克隆质粒进行鉴定 ,然后进行序列测定 ,证实为S1基因。将此重组质粒命名为pMDJS950 3S。     

Enzootic bovine leukosis application of E.L.I.S.A. technique for the detection of antibodies

A simple microplate method of enzyme-linked immunosorbent assay (E.L.I.S.A.) for detecting antibodies to bovine leukosis virus (B.L.V.) is described. The antigen consisted of a solution containing the two major antigens of the B.L.V. (gp 51 and p 24) obtained by a technique of purification using CN-Br activated Sepharose 4B. This E.L.I.S.A. was compared with the agar gel immunodiffusion test (A.G.I.D.T.) in a study of 545 bovine sera. The total discrepancy rate between the two tests was 11% with a better sensitivity for E.L.I.S.A.