Serotyping, molecular typing, and vaccine protein screening for Riemerella anatipestifer: Overcoming challenges in...

Riemerella anatipestifer (R. anatipestifer) affects the duck farming industry worldwide, causing substantial economic losses. The current disease prevention and treatment strategies primarily include vaccines and antibiotics. However, the large number of serotypes and increasing resistance to R. anatipestifer make it challenging to prevent and treat the infection. This study carried out the serotyping and molecular typing of 51 R. anatipestifer strains and predicted vaccine proteins based on pan-genome analysis and cross-immune protection potential. For serotype identification, the rabbits were immunized with antigens, and 9 serotyped sera were prepared, the data revealed 6 serotypes with two unformed strains. The results for the self-made serotypes were consistent with those obtained from the externally submitted strains. Moreover, the pan-genome analysis was performed on 51 R. anatipestifer strains, and an open pan-genome set of 5094 genes was constructed. In addition, the COG annotation classification indicated that the core and non-core genomes were significantly different in gene functions. A total of 1116 core genomes that could serve as better cross-protective vaccine proteins were analyzed and revealed 5 genes of interest. In addition, the oprM-1 protein, a highly reactive protein, was expressed and purified, and the immunoreactivity with five antisera (anti-serotypes 1, 2, 5, 11, and 18) was demonstrated by Western blotting. This study fills the gaps in the existing typing systems for R. anatipestifer by combining serotyping, MLST typing, and pan-genome analysis. Furthermore, it provides valuable insights into the epidemiology, evolution, and pathogenesis of R. anatipestifer and paves the way for developing effective cross-protective vaccines.