鸡肠炎沙门氏菌ompA基因的克隆与表达

根据GenBank中鸡肠炎沙门氏菌ompA基因序列设计1对引物,以鸡肠炎沙门氏菌贵州分离株基因组为模板,扩增鸡肠炎沙门氏菌ompA基因,将其亚克隆到原核表达载体pET-32a(+)中,构建重组表达质粒pET-32a-ompA,转化大肠杆菌BL21(DE3)感受态细胞, 经IPTG诱导, 实现了鸡肠炎沙门氏菌ompA蛋白在大肠杆菌中的表达。SDS-PAGE分析结果表明, 该重组蛋白的分子质量约为55 ku,可溶性分析结果表明表达蛋白大部分以包涵体形式存在。Western blotting分析结果表明,该重组蛋白具备免疫原性。     

Comparative Pathogenesis of Experimental Infections with Salmonella gallinarum in Local and Commercial Chickens

The pathogenicity of a virulent strain of Salmonella gallinarum was studied in local chickens and commercial layers. Sixty 4-month-old chickens were used. Of these, 50 chickens were orally infected with S. gallinarum, comprising 25 commercial layers and 25 local chickens. Five chickens in each group were used as uninfected negative controls. The clinical signs and pathological features of acute, subacute and chronic fowl typhoid were observed in both groups. Chickens in both groups seroconverted, but the antibody titre was significantly higher (p<0.001) in the commercial layers. The antibody titre remained high to the end of the experiment in all the surviving chickens. Only one commercial layer chicken died during the course of the experiment. The PCV decreased significantly (p<0.001) in the infected chickens of both groups as compared to the controls. The viable cell count of S. gallinarum in the liver and spleen reached a maximum on day 9 after infection in both groups. However, there was a significantly higher cell count (p<0.05) in the commercial layers. The severity of the disease appeared to be slightly greater in the commercial layers, although the susceptibility was similar in both groups. It was concluded that, under experimental conditions, local chickens are just as susceptible to S. gallinarum infection as are commercial layers.     

The role of rpoS,hmp, and ssrAB in Salmonella enterica Gallinarum and evaluation of a triple-deletion mutant as a live vaccine candidate in Lohmann layer chickens

Salmonella enterica Gallinarum (SG) causes fowl typhoid (FT), a septicemic disease in avian species. We constructed deletion mutants lacking the stress sigma factor RpoS, the nitric oxide (NO)-detoxifying flavohemoglobin Hmp, and the SsrA/SsrB regulator to confirm the functions of these factors in SG. All gene products were fully functional in wild-type (WT) SG whereas mutants harboring single mutations or a combination of rpoS, hmp, and ssrAB mutations showed hypersusceptibility to H₂O₂, loss of NO metabolism, and absence of Salmonella pathogenicity island (SPI)-2 expression, respectively. A triple-deletion mutant, SGΔ3 (SGΔrpoSΔhmpΔssrAB), was evaluated for attenuated virulence and protection efficacy in two-week-old Lohmann layer chickens. The SGΔ3 mutant did not cause any mortality after inoculation with either 1 × 10⁶ or 1 × 10⁸ colony-forming units (CFUs) of bacteria. Significantly lower numbers of salmonellae were recovered from the liver and spleen of chickens inoculated with the SGΔ3 mutant compared to chickens inoculated with WT SG. Vaccination with the SGΔ3 mutant conferred complete protection against challenge with virulent SG on the chickens comparable to the group vaccinated with a conventional vaccine strain, SG9R. Overall, these results indicate that SGΔ3 could be a promising candidate for a live Salmonella vaccine against FT.