Characterization and Evaluation of a Salmonella enterica Serotype Senftenberg Mutant Created by Deletion of Virulence-Related Genes for Use as a Live Attenuated Vaccine.

Natural infections of chickens with Salmonella enterica subsp. enterica serovar Senftenberg (S. Senftenberg) are characterized by low-level intestinal invasiveness and insignificant production of antibodies. In this study, we investigated the potential effects of lon and cpxR gene deletions on the invasiveness of S Senftenberg into the intestinal epithelium of chickens and its ability to induce an immune response, conferring protection against S Senftenberg infection. With the allelic exchange method, we developed JOL1596 (Δlon), JOL1571 (ΔcpxR), and JOL1587 (Δlon ΔcpxR) deletion mutants from wild-type S Senftenberg. Deletion of the lon gene from S Senftenberg produced increased frequency of elongated cells, with significantly greater amounts of exopolysaccharide (EPS) than in the cpxR-deleted strain and the wild-type strain. The in vivo intestinal loop invasion assay showed a significant increase in epithelial invasiveness for JOL1596 (Δlon) and JOL1587 (Δlon ΔcpxR), compared to JOL1571 (ΔcpxR) and the wild-type strain. Furthermore, the S Senftenberg wild-type and mutant strains were internalized at high levels inside activated abdominal macrophages from chicken. The in vivo inoculation of JOL1587 (Δlon ΔcpxR) into chickens led to colonization of the liver, spleen, and cecum for a short time. Chickens inoculated with JOL1587 (Δlon ΔcpxR) showed significant increases in humoral, mucosal, and cellular immune responses specific to S Senftenberg antigens. Postchallenge, compared to the control group, the JOL1587 (Δlon ΔcpxR)-inoculated chickens showed not only lower persistence but also faster clearance of wild-type S Senftenberg from the cecum. We conclude that the increased intestinal invasiveness and colonization of internal organs exhibited by JOL1587 (Δlon ΔcpxR) led to the establishment of immunogenicity and conferred protective efficacy against S Senftenberg infections in chickens.