Identification and Characterization of Biosynthetic Loci of Lipooligosaccharide and Capsular Polysaccharide in Avibacterium paragallinarum

Abstract

Infectious coryza is an acute respiratory disease in chickens caused by Avibacterium paragallinarum. Lipooligosaccharides (LOSs) and capsular polysaccharides are important components of Av. paragallinarum. Herein, we identified that gene cluster L6 and two genes waaF, waaQ were associated with LOS synthesis, and two genes acbD and ccbF1 were involved in capsular synthesis. Mutant and complementary strains of these genes were generated by natural transformation. Wild-type strains produced LOS that yielded an upper and lower band. In comparison, ΔwaaQ and ΔwaaF yielded a truncated lower band and lacked the upper band, while ΔL6 did not exhibit the upper band, and the lower band was identical to that of the wild-type strain. The survival rates of wild-type strain, ΔwaaF, ΔwaaQ, and ΔL6 in chicken serum were 4.89% ± 0.27%, 0.0013% ± 0.0002%, 0.43% ± 0.05%, and 3.1% ± 0.35%, respectively. Notably, the resistances of ΔwaaF, ΔwaaQ, and ΔL6 to chicken serum were significantly lower than that of parent strain. By contrast, the survival rate of the ΔacbD strain was 55.17% ± 0.61%, and its resistance to chicken serum was significantly higher than that of the wild-type strain (p < 0.001). Deletion of the waaF, waaQ, L6, acbD, and ccbF1 genes resulted in enhanced formation of biofilm without altering immunogenicity in chickens. The ΔwaaF, ΔwaaQ, and ΔccbF1 strains exhibited heightened susceptibility to fowlicidin-2. Furthermore, ΔwaaF, ΔacbD, and ΔccbF1 strains shown a decrease in pathogenicity (p < 0.05). These results are valuable for advancing research on the pathogenesis of Av. paragallinarum.