Abstract
Salmonellosis is a major public health problem throughout the world. Thus, there is a huge need for diversified control strategies for Salmonella infections. In this work, we have assessed the potential use of Bacillus subtilis (B. subtilis) spores for the expression of a major protective antigen of Salmonella serovar Pullorum, OmpC. The expression of OmpC on the surface of spores was determined by immunofluorescence microscopy. Mice immunized with recombinant spores expressing the OmpC antigen presented significant levels of OmpC-specific serum IgG and mucosal SIgA antibodies than in mice immunized with non-recombinant spores (p<0.01). In addition, oral immunization with recombinant spores was able to induce a significant level of protection in mice against lethal challenge with Salmonella serovar Typhimurium. These results suggest that B. subtilis spores have promising potential in the development of mucosal vaccines against Salmonella infections.
Highlights
Pullorum disease (PD) is a worldwide poultry disease that caused enormous economic losses throughout large areas of the world
To obtain recombinant B. subtilis spores expressing OmpC on their surface, a recombinant plasmid pDG364-cotC-ompC containing the cotC::ompC gene fusion for double cross-over with B. subtilis chromosome was constructed by fusing the ompC gene into frame of the coding part of cotC gene (Fig 1A)
CotC::ompC gene fusion was integrated into the B. subtilis chromosome at the nonessential amyE gene locus by double cross-over event (Fig 1B)
Summary
Pullorum disease (PD) is a worldwide poultry disease that caused enormous economic losses throughout large areas of the world. Different strategies to control Salmonella infections are currently available, continuous emergence of multi-drug resistance [3, 4] and novel Salmonella variants [5, 6] are still a mortal threat to the poultry industry and public health. Since most pathogenic organisms, including Salmonella, penetrate to the host through the mucosal membranes, effective mucosal vaccines that induce immunity at the site of infection are able to confer more efficient protective immune responses against Salmonella [7, 8]. There is much evidence that OmpC, an outer membrane protein (porin) from Salmonella, is a promising candidate antigen that efficiently stimulates innate and adaptive immune responses
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