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Avian pathogenic Escherichia coli (APEC) causes avian airsacculitis and colibacillosis, resulting in significant economic loss to the poultry industry. O1, O2 and O78 are the three predominant serotypes. O-antigen of lipopolysaccharide is serotype determinant and highly immunogenic, and O-antigen polysaccharide-based vaccines have great potential for preventing bacterial infections. In this study, we utilized a novel yeast/bacterial shuttle vector pSS26 to clone the 10.8 kb operon synthesizing APEC O1 O-antigen polysaccharide. The resulting plasmid was introduced into attenuated Salmonella vaccines to deliver this O-antigen polysaccharide. O1 O-antigen was stably synthesized in attenuated Salmonella Typhimurium, demonstrated by slide agglutination, silver staining and western blot. Our results also showed that APEC O1 O-antigen produced in the Salmonella vaccines was attached to bacterial cell surfaces, and the presence of heterologous O-antigen did not alter the resistance to surface-acting agents. Furthermore, birds immunized orally or intramuscularly provided protection against the virulent O1 APEC challenge. Salmonella vaccines carrying APEC O1 O-antigen gene cluster also induced high IgG and IgA immune responses against lipopolysaccharide from the APEC O1 strain. The use of our novel shuttle vector facilitates cloning of large DNA fragments, and this strategy could pave the way for production of Salmonella-vectored vaccines against prevalent APEC serotypes.

pathogens and disease

A biologically conjugated polysaccharide vaccine delivered by attenuated Salmonella Typhimurium provides protection against challenge of avian pathogenic Escherichia coli O1 infection