牛结核病的防控

牛结核病是由结核分枝杆菌引起的一种人畜共患慢性传染病。为有效控制和净化牛结核病,切实保障畜牧业生产安全、动物产品质量安全和公共卫生安全,必须坚持预防为主、因地制宜、分类指导、逐步净化的防控方针和策略,把养殖场（户）作为防控本病的主体,不断完善奶牛养殖场生物安全体系,严格落实监测净化、检疫监管、无害化处理、应急处置等综合防控措施,积极开展场群和区域净化工作,有效清除病原,降低发病率,压缩流行范围,逐步实现防控工作总体达标。     

Chronic tuberculosis caused by Mycobacterium bovis in a domestic donkey in Central Europe

Bovine tuberculosis is a contagious and zoonotic disease of animals and humans. In Europe, the number of reported cases of tuberculosis has decreased. Equidae are relatively rarely infected even in endemic areas. The presented report describes a case of chronic Mycobacterium bovis tuberculosis in a 30-year-old female donkey. The donkey initially presented with persistent lymphadenopathy; however, as the disease progressed, weight loss became apparent. To the authors' knowledge, this is the second confirmed case of tuberculosis in a donkey in Europe.     

Tuberculosis vaccination sequence effect on protection in wild boar

The Eurasian wild boar (Sus scrofa) is a reservoir for tuberculosis (TB) in which vaccination is a valuable tool for control. We evaluated the protection and immune response achieved by homologous and heterologous regimes administering BCG and heat-inactivated Mycobacterium bovis (IV). Twenty-one wild boar piglets were randomly allocated in five groups: Control, homologous BCG, homologous IV, heterologous IV-BCG, heterologous BCG-IV. Significant 67% and 66% total lesion score reductions were detected in homologous IV (IVx2) and heterologous IV-BCG groups when compared with Control group (F_4,16 = 6.393, p = 0.003; Bonferroni _{Control vs IVx2} p = 0.026, Tukey _{Control vs IV-BCG} p = 0.021). No significant differences were found for homologous BCG (although a 48% reduction in total lesion score was recorded) and BCG-IV (3% reduction). Heterologous regimes did not improve protection over homologous regimes in the wild boar model and showed variable results from no protection to similar protection as homologous regimes. Therefore, homologous regimes remain the best option to vaccinate wild boar against TB. Moreover, vaccine sequence dramatically influenced the outcome underlining the relevance of studying the effects of prior sensitization in the outcome of vaccination.     

Expression,Purification and Activity Evaluation of Mb1230 Gene of Mycobacterium bovis

To explore the value of Mb1230 protein of Mycobacterium bovis in the diagnosis of bovine tuberculosis,we obtained the Mb1230 gene by PCR and constructed recombinant plasmid pET-22b-Mb1230.Recombinant Mb1230 protein was obtained by IPTG induction and purified by affinity chromatography.The activity of the recombinant protein was evaluated by TST test,IGRA test and indirect ELISA.The size of the recombinant protein matched with the theoretical value proved by SDS-PAGE;Western blotting result showed that the recombinant protein could react with mouse anti-His antibody,and had specific band;The results of TST test,IGRA test and indirect ELISA test also showed the recombinant protein had antigenic activity.The results indicated the recombinant protein Mb1230 had good B cell and T cell activity,so,it had the potential application in the diagnosis of bovine tuberculosis.     

抑制鹿源多药耐药结核菌中药的筛选

为了筛选对鹿源多药耐药结核菌有效的中药耐药抑制剂,通过MABA法检测24种中药的水和乙醇提取物对多药耐药结核菌及标准菌H37Rv的体外抗菌效果,将筛选出的抗菌效果好的黄连醇提物和黄连水提物采用紫外分光光度法进行活性成分含量测定,同时以MABA法与标准品比较体外抗结核菌活性。结果显示,黄连、独活和侧柏叶有明显的抗结核活性,其中黄连水粗提物和乙醇粗提物效果最佳,测得它们活性成分即总生物碱含量分别为22.74%和35.23%,并显示二者与小檗碱的体外抗结核作用相同。     

结核分枝杆菌蛋白TB9.7间接ELISA方法的建立

为获得新的鉴别诊断结核杆菌感染的候选抗原,根据Gene Bank中登录的结核杆菌H37Rv的TB 9.7基因序列设计1对引物,以结核杆菌H37Rv基因组DNA为模板,PCR扩增得到TB 9.7基因DNA片段。将该扩增片段克隆于PMD18-T载体中,得到载体PMD18-T-TB9.7。分别将pET32a质粒和PMD18-T-TB 9.7质粒用限制性内切酶EcoRⅠ和XhoⅠ双酶切,并将纯化的TB 9.7基因亚克隆至pET32a中,获得重组表达质粒pET32a-TB 9.7。将其转化E.coli BL21 IPTG诱导后,经SDS-PAGE、Western blot分析鉴定,可见约30ku的外源蛋白带。表明TB9.7基因在大肠杆菌中得到了表达。用Ni-NTA Agarose试剂盒进行蛋白纯化,获得纯化的TB9.7融合蛋白,建立间接ELISA诊断方法,检测了30份结核病人的痰检阳性血清,120份健康人的血清,敏感性为56.6%,特异性为97%,与痰检结果的符合率为84%。     

牛分枝杆菌Mb1230基因的表达纯化及其活性评价

为了探究牛分枝杆菌Mb1230蛋白在牛结核病诊断中的应用价值,本试验利用PCR方法扩增出Mb1230基因,构建重组质粒pET-22b-Mb1230,经IPTG诱导表达后用亲和层析纯化重组蛋白,通过结核菌素皮内变态反应试验(TST试验)、IFN-γ释放试验(IGRA试验)和间接ELISA对重组蛋白的活性进行评价。SDS-PAGE和Western blotting结果显示,重组蛋白大小与理论值相符,且能与鼠抗His的抗体反应有特异性条带;在TST试验、IGRA试验和间接ELISA中也表现出抗原活性。结果表明,重组Mb1230蛋白具有良好的B细胞活性和T细胞活性,在牛结核病诊断中具有应用潜力。     

,y康汤对复治性肺结核阴虚毒癖证疗效及 T淋巴细胞的影响

〔摘要〕目的研究傍康汤对复治性肺结核阴虚毒疲证患者T淋巴细胞的影响、方法将%例阴虚毒疲型复治 性肺结核患者随机分为两组:治疗组48例和对照组48例,并设立正常组48例;对照组采用西医常规治疗,治疗组 在西医常规治疗基础上加服傍康汤,2周为1个疗程,共治疗12个疗程〔观察两组临床疗效及对CD,'',CD,'',CDH''1'' 淋巴细胞、CD,''/CDH+比值的影响〔结果治疗组总有效率为89.60Io,与对照组66.7%比较,差异有显著统计学意义(P< 0.01);两组CD,'',CDH''1''淋巴细胞、C D,''/C DH+比值治疗前与正常组比较有显著差异}P<0.01),治疗后与治疗前比较 C D,'' T淋巴细胞、CD,''/CDH+比值明显升高,CDH'' T淋巴细胞则明显卜降,差异有统计学意义(P<0.05 ) ;治疗后治疗组 CD,'',CD,''/CDH''比值的升高、CDH''的h降均优于对照组(P<0.05 )〔结论傍康汤治疗阴虚毒疲型复治性肺结核疗效确 切,其机制可能是通过改善患者T淋巴细胞免疫功能而起作用、     

牛结核分枝杆菌溶血磷脂酶真核表达载体的构建及在Hela细胞中的表达

为了研究牛结核分枝杆菌溶血磷脂酶基因(Lip)的生物学特性及其功能,试验构建了Lip-pEGFP融合基因的真核表达载体,利用脂质体(lipofectamine2000)将其转染到Hela细胞中,用荧光显微镜检测GFP在细胞中的表达情况。结果显示,重组Lip-pEGFP载体在Hela细胞中获得了高效表达,且能够稳定传代,为进一步研究Lip的生物学功能及其在牛结核分枝杆菌中的作用奠定了基础。     

分枝杆菌阿拉伯半乳聚糖生物合成途径研究进展

综述了阿拉伯半乳聚糖生物合成关键基因、合成步骤及其调控因素的研究进展,利用结核分枝杆菌的基因组序列,从比较基因组层面分析了阿拉伯半乳聚糖生物合成的保守性,为利用耻垢分枝杆菌模型筛选阿拉伯半乳聚糖合成过程中潜在的药物作用靶标提供了基础。