Search

Brucella poses a great threat to animal and human health, and vaccination is a good way of controlling the bacterium. In this study, the immune response and protection ability of a recombinant adenovirus Ad-LL/BP containing L7/L12 and BCSP31 proteins of Brucella abortus (B. abortus) were evaluated in BALB/c mice model. Firstly, Adenovirus vector Ad-CMV and the recombinant adenovirus Ad-LL/BP were amplified in HEK 293AD cells. The TCID50 values of Ad-LL/BP and Ad-CMV were 10−8.68/0.1 mL and 10−8.35/0.1 mL, respectively. Mice were inoculated with 100 TCID50/mouse of Ad-LL/BP or Ad-CMV. Vaccination of mice with Ad-LL/BP vaccine was able to elicit higher IgG, IgG1 and IgG2a antibody levels when compared with Ad-CMV and PBS control animals (P0.05). Ad-LL/BP vaccine was able to reduce significantly the numbers of B. abortus in the spleens from the immunized mice. These results indicated that the recombinant Ad-LL/BP vaccine induced mainly cell-mediated immunity and partly humoral immunity, and provided moderately protection against B. abortus infection. Therefore, the vaccine could be further developed into a live-vector vaccine against B. abortus.

This study identified and characterized Brucella species in the informal milk chain in Uganda. A total of 324 cattle bulk milk samples were screened for the genus Brucella by real-time PCR with primers targeting the bcsp31 gene and further characterized by the omp25 gene. Of the samples tested, 6.5% were positive for Brucella species. In the omp25 phylogeny, the study sequences were found to form a separate clade within the branch containing B. abortus sequences. The study shows that informally marketed cattle milk in Uganda is a likely risk factor for human brucellosis and confirms that B. abortus is present in the cattle population. This information is important for potential future control measures, such as vaccination of cattle.

Background & Objective: One of the most common diseases between zoonosis - especially in developing countries – is brucellosis. Identification of Brucella cell antigen combinations in terms of the amount and type of immune response in infected hosts, are important in vaccine design. 31kDa Brucella cell surface protein (BCSP31) is shared among all Brucellae. We aimed to define specific immune responses to BCSP31 which are elicited during infection of murine host with B. abortus. Surface protein of 31 kDa (BCSP31) is present in all strains, and here, the host immune response during murine infection with Brucella abortus which are formed in proportion to the proteins are studied. Materials & Methods: Recombinant BCSP31 (rBCSP31) of B. abortus was produced and purified. One group of BALB/c mice were infected with pathogenic B. abortus 544 and maintained for 60 days; a no-infected group of mice also was considered. Specific serum antibodies directed to rBCSP31 was evaluated through ELISA. Splenocytes of mice were cultured and stimulated with rBCSP31; thereafter, IL-4, IL-12 and IFN-γ cytokine responses of spleen lymphocytes were assessed. Results: We detected a remarkable IgG titer along with IgG1 and IgG2a specific to recombinant BCSP31 in serum samples from infected mice. Significant titers of IgG, IgG1 and IgG2a antibodies than BCSP31 in the the serum of mice infected with the virulent strain were observed. The Evaluation of Splenocytes responses to rBCSP31 also showed a significant IL-12 and IFN-γ production along with remarkable IL-4 production in infected mice. All responses were significantly different from that of non-infected mice (p

Several pathogens including Brucella spp. are shed in semen of infected bulls and can be transmitted to cows through contaminated semen during artificial insemination. The present study reports omp2a and bcsp31 gene based loop-mediated isothermal amplification (LAMP) assays for detection of Brucella genomic DNA in semen from infected bulls. The positive results could be interpreted visually by change in colour of reaction mixture containing hydroxyl naphthol blue (HNB) dye from violet to sky blue. LAMP assays based on omp2a and bcsp31 could detect as little as 10 and 100 fg of B. abortus S19 genomic DNA, respectively. Sensitivity of omp2a and bcsp31 LAMP assays for direct detection of organisms in bovine semen was 2.28 × 10 CFU and 2.28 × 10 CFU of B. abortus S19 in spiked bovine semen, respectively. The omp2a LAMP assay was found equally sensitive to TaqMan probe based real-time PCR and 100 times more sensitive than conventional PCR in identifying Brucella in spiked semen. The diagnostic applicability of the omp2a LAMP assay was evaluated with seventy-nine bovine semen samples and results were re-evaluated through TaqMan probe based real-time PCR and conventional PCR. Taken together, the omp2a LAMP assay is easy to perform, rapid and sensitive in diagnosis of Brucella spp. in bovine semen. [ABSTRACT FROM AUTHOR]

This study identified and characterized Brucella species in the informal milk chain in Uganda. A total of 324 cattle bulk milk samples were screened for the genus Brucella by real-time PCR with primers targeting the bcsp31 gene and further characterized by the omp25 gene. Of the samples tested, 6.5% were positive for Brucella species. In the omp25 phylogeny, the study sequences were found to form a separate clade within the branch containing B. abortus sequences. The study shows that informally marketed cattle milk in Uganda is a likely risk factor for human brucellosis and confirms that B. abortus is present in the cattle population. This information is important for potential future control measures, such as vaccination of cattle. [ABSTRACT FROM AUTHOR]

Background & Objective: One of the most common diseases between zoonosis - especially in developing countries -- is brucellosis. Identification of Brucella cell antigen combinations in terms of the amount and type of immune response in infected hosts, are important in vaccine design. 31kDa Brucella cell surface protein (BCSP31) is shared among all Brucellae. We aimed to define specific immune responses to BCSP31 which are elicited during infection of murine host with B. abortus. Surface protein of 31 kDa (BCSP31) is present in all strains, and here, the host immune response during murine infection with Brucella abortus which are formed in proportion to the proteins are studied. Materials & Methods: Recombinant BCSP31 (rBCSP31) of B. abortus was produced and purified. One group of BALB/c mice were infected with pathogenic B. abortus 544 and maintained for 60 days; a no-infected group of mice also was considered. Specific serum antibodies directed to rBCSP31 was evaluated through ELISA. Splenocytes of mice were cultured and stimulated with rBCSP31; thereafter, IL-4, IL-12 and IFN-γ cytokine responses of spleen lymphocytes were assessed. Results: We detected a remarkable IgG titer along with IgG1 and IgG2a specific to recombinant BCSP31 in serum samples from infected mice. Significant titers of IgG, IgG1 and IgG2a antibodies than BCSP31 in the the serum of mice infected with the virulent strain were observed. The Evaluation of Splenocytes responses to rBCSP31 also showed a significant IL-12 and IFN-γ production along with remarkable IL-4 production in infected mice. All responses were significantly different from that of non-infected mice (p<0.05). Conclusion: These findings suggest that specific humoral and cell-mediated responses to BCSP31 is formed during murine host infection with B. abortus. Based on these findings, rBCSP31 can be used in further design of immunogenic strategies for vaccination against brucellosis. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop a LightCycler-based real-time PCR (LC-PCR) assay and to evaluate its diagnostic use for the detection of Brucella DNA in serum samples. Following amplification of a 223-bp gene sequence encoding an immunogenetic membrane protein (BCSP31) specific for the Brucella genus, melting curve and DNA sequencing analysis was performed to verify the specificity of the PCR products. The intra- and inter-assay variation coefficients were 1.3% and 6.4%, respectively, and the detection limit was 5 fg of Brucella DNA (one genome equivalent). After optimisation of the PCR assay conditions, a standard curve was obtained with a linear range (correlation coefficient = 0.99) over seven orders of magnitude from 107 to 10 fg of Brucella DNA. The LC-PCR assay was found to be 91.9% sensitive and 95.4% specific when tested with 65 negative control samples and 62 serum samples from 60 consecutive patients with active brucellosis. The assay is reproducible, easily standardised, minimises the risk of infection in laboratory workers, and has a total processing time of < 2 h. It could therefore form a promising and practical approach for the rapid diagnosis of human brucellosis. [ABSTRACT FROM AUTHOR]

The article discusses a study that examined the molecular characterization of the contagious disease Brucella abortus from cattle and buffaloes. Topics covered include the inoculation of 84 samples of vaginal mucus, placenta, cotyledons and fetal stomach contents from aborted cattle and buffaloes using Brain Heart Infusion as the selective media, and the effectiveness of the use of polymerase chain reaction in the diagnosis of brucellosis.