Identification of a gene in Riemerella anatipestifer CH-1 (B739-2187) that contributes to resistance to polymyxin B and evaluation of its mutant as a live attenuated vaccine

• Gene B739-2187 of Riemerella anatipestifer CH-1 has ability to confer Salmonella ΔphoP mutant to resistance to polymyxin B.
• B739-2187 contributed to resistance of R. anatipestifer CH-1 to polymyxin B.
• Disruption of B739-2187 in R. anatipestifer CH-1 attenuated its virulence.
• The mutant ΔB739-2187 has potential for developing live attenuated vaccine against infection of R. anatipestifer.

Riemerella anatipestifer (R. anatipestifer) causes severe perihepatitis, pericarditis, airsacculitis and meningitis in the duck, leading to great economic losses worldwide. Given the increased prevalence of drug-resistance strains, vaccination is the best strategy to prevent R. anatipestifer infection in ducklings. In this study, we identified a gene in R. anatipestifer (B739-2187) that can restore the resistance of the Salmonella phoP mutant to polymyxin B using genetic complementation. Furthermore, the deletion of B739-2187 in R. anatipestifer resulted in a mutant exhibiting increased sensitivity to polymyxin B. The R. anatipestifer B739-2187 mutant did not exhibit phenotypic defects, as indicated by its growth curve, lipopolysaccharide and outer membrane protein profiles, and attachment and invasion of duck embryo fibroblast cells. The duck animal experiments demonstrated that the deletion of B739-2187 significantly decreased the virulence of R. anatipestifer, and the B739-2187 mutant provided 100% protection against challenge with wild-type R. anatipestifer, exhibiting the characteristics of an ideal live vaccine.