Protective efficacy by various doses of a new brucellosis vaccine candidate based on Salmonella strains expressing Brucella abortus BSCP31, Omp3b and superoxide dismutase against brucellosis in murine model

Brucella species are important etiological agents of zoonotic diseases. Attenuated Salmonella strains expressing Brucella abortus BCSP31, Omp3b and superoxide dismutase proteins were tested as vaccine candidates in this study. In order to determine the optimal dose for intraperitoneal (IP) inoculation required to obtain effective protection against brucellosis, mice were immunized with various doses of a mixture of the three vaccine strains. Fifty BALB/c mice were divided into five equal groups (groups A-E). Group A mice were intraperitoneally inoculated with 100 μL of sterile phosphate-buffered saline. Group B, C, D and E mice were intraperitoneally immunized with approximately 1.2 × 10⁵ colony-forming units (CFU) mL^-1 of Salmonella containing pMMP65 in 100 μL andwith 1.2 × 10⁴ CFU mL^-1, 1.2 × 10⁵ CFU mL^-1 and 1.2 × 10⁶ CFU mL^-1 of the mixture of the three strains in 100 μL, respectively. Serum IgG, tumor necrosis factor alpha and interferon gamma concentrations were significantly higher in group E than in groups A-D. Following challenge with B. abortus 544, the challenge strain was not detected in the spleen of any mouse from group E. Thus, IP immunization with 1.2 × 10⁶ CFU mL^-1 of the mixture of the three vaccine strains induced immune responses and provided effective protection against brucellosis in mice.