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.     

结核分枝杆菌分泌蛋白ESAT-6、CFP10基因的克隆、鉴定及其原核表达

以人型结核分枝杆菌 H37RV株基因组 DNA为模板 ,应用 PCR法对 ESAT- 6和 CFP10基因进行扩增 ,产物经纯化后与载体 PMD18- T连接、转化及酶切鉴定 ,亚克隆到原核表达载体 PGEX- 6 P- 1,构建原核重组表达质粒 ,转化入大肠杆菌 BL2 1中 ,以 1mmol/L IPTG诱导 ,进行 SDS- PAGE电泳。结果表明 ,ESAT- 6和 CFP10基因表达的融合蛋白相对分子质量分别为 32 0 0 0和 36 0 0 0 ,与实测相符。重组结核杆菌分泌蛋白 ESAT- 6和 CFP10的成功表达为结核病诊断及重组疫苗的构建打下了基础     

Multisystemic bovine mycobacteriosis in a pony with neurological signs and weight loss

This report describes the clinical, biochemical and histopathological findings of a case of disseminated, primary mycobacterial infection with Mycobacterium bovis in a pony presenting with neurological signs and weight loss. This report revisits tuberculosis as a potential differential diagnosis in horses at a time of relatively strong tuberculosis control in the UK and iterates the importance of post‐mortem examination as a diagnostic tool for clinicians. Key public health questions following exposure to mycobacterial pathogens are also discussed.     

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.     

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.     

Simulation model of within-herd transmission of bovine tuberculosis in Argentine dairy herds

Transmission of bovine tuberculosis was quantified in three dairy herds located in south Santa Fe Province, Argentina. Using estimates of Mycobacterium bovis transmission (β) and a Reed–Frost simulation model, the prevalence of tuberculosis infection in the study herds over time was investigated. The Reed–Frost model was modified by incorporating randomness in both β and the incubation period () of M. bovis. The mean estimated herd β was 2.2 infective contacts per year and did not differ significantly between the study herds. Modeling as Poisson distributed (mean 24 months) best fit the observed prevalences. Infection was predicted by the model either to spread quickly (<10 years) within a herd and reach a high prevalence (>50%), or to persist at a low prevalence (<5–10%). The model was robust, predictions were realistic and the mean β estimated was consistent with previous studies of bovine tuberculosis.     

Evaluation of the Abbott LCx Mycobacterium Tuberculosis Assay for Direct Detection of Mycobacterium Bovis in Bovine Tissue Samples

The commercial LCx amplification assay, usually employed to detect the Myocobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using Löwenstein-Jensen solid medium and pathological findings on 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. Fifty-three cultures (51 and 2, respectively) were positive for M. bovis, while the results for the LCx assay and the histological method were positive in 48 (45, 3) and 24 (20, 4) samples, respectively. None of the samples from cattle from certified tuberculosis-free herds were positive by any of the procedures. The results obtained with the LCx assay, compared with the culture procedure, regarded as the gold standard among the diagnostic techniques, gave a specificity of 91.6% and sensitivity of 90.5%. Although the sensitivity of LCx was suboptimal, DNA of M. bovis was detected in 81.8% of the skin test-positive animals. Amplification techniques could provide a rapid and reasonably reliable tool for detecting bovine tuberculosis.     

Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

牛分支杆菌CFP-10的原核表达及其在牛结核病诊断中的应用

低分子质量的蛋白抗原CFP-10是一种重要的牛分支杆菌早期分泌蛋白.为了检测该蛋白和其他几种抗原在牛结核病诊断中的临床应用,对CFP-10的基因进行克隆,鉴定,并在原核系统中表达.PCR法扩增cfp-10基因片段,连接到pET-22b( )原核表达载体中,再转入表达宿主E.coli BL21(DE3)PlysS菌株内,用IPTG诱导,进行蛋白表达、纯化.分别以CFP-10、ESAT-6、MPT83、MPT70、牛PPD、CFP-10与ESAT-6混合蛋白,MPT83与MPT70混合蛋白为抗原,用间接ELISA法诊断牛结核病.结果表明,以CFP-10与ESAT-6混合蛋白作为抗原检测牛结核病的特异性和敏感性分别达到了100%和63.6%,均超过了其他单抗原或抗原组合,为以筛选合适抗原为基础的血清学诊断技术提供了有力的支持并创造了良好的应用前景.     

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。