Boyd, Jessica M., Dacanay, Andrew, Knickle, Leah C., Touhami, Ahmed, Brown, Laura L., Jericho, Manfred H., Johnson, Stewart C., and Reith, Michael
ABSTRACTAeromonas salmonicidasubsp. salmonicida, a bacterial pathogen of Atlantic salmon, has no visible pili, yet its genome contains genes for three type IV pilus systems. One system, Tap, is similar to the Pseudomonas aeruginosaPil system, and a second, Flp, resembles the Actinobacillus actinomycetemcomitansFlp pilus, while the third has homology to the mannose-sensitive hemagglutinin pilus of Vibrio cholerae. The latter system is likely nonfunctional since eight genes, including the gene encoding the main pilin subunit, are deleted compared with the orthologous V. choleraelocus. The first two systems were characterized to investigate their expression and role in pathogenesis. The pili of A. salmonicidasubsp. salmonicidawere imaged using atomic force microscopy and Tap- and Flp-overexpressing strains. The Tap pili appeared to be polar, while the Flp pili appeared to be peritrichous. Strains deficient in tapand/or flpwere used in live bacterial challenges of Atlantic salmon, which showed that the Tap pilus made a moderate contribution to virulence, while the Flp pilus made little or no contribution. Delivery of the tapmutant by immersion resulted in reduced cumulative morbidity compared with the cumulative morbidity observed with the wild-type strain; however, delivery by intraperitoneal injection resulted in cumulative morbidity similar to that of the wild type. Unlike the pili of other piliated bacterial pathogens, A. salmonicidasubsp. salmonicidatype IV pili are not absolutely required for virulence in Atlantic salmon. Significant differences in the behavior of the two mutant strains indicated that the two pilus systems are not redundant.