Evidence for horizontal gene transfer, gene duplication and genetic variation as driving forces of the diversity of haemolytic phenotypes in Photobacterium damselae subsp. damselae

Photobacterium damselae subsp. damselae , a marine bacterium that causes infections in marine animals and in humans, produces up to three different haemolysins involved in virulence, which include the pPHDD1 plasmid-encoded damselysin (Dly) and HlyApl, and the chromosome-encoded HlyAch. We screened 45 isolates from different origins, and found a correlation between their haemolytic phenotypes and the differential haemolysin gene content. All highly and medium haemolytic strains harboured pPHDD1, with amino acid substitutions in HlyApl and HlyAch being the cause of the medium haemolytic phenotypes in some pPHDD1-harbouring strains. Weakly haemolytic strains contained only hlyA ch , whereas nonhaemolytic isolates, in addition to lacking pPHDD1, either lacked hlyA ch or contained a hlyA ch pseudogene. Sequence analysis of the genomic context of hlyA ch uncovered an unexpected genetic diversity, suggesting that hlyA ch is located in an unstable chromosomal region. Phylogenetic analysis suggested that hlyA pl and hlyA ch originated by gene duplication within P. damselae subsp. damselae following acquisition by horizontal transfer. These observations together with the differential distribution of pPHDD1 plasmid among strains suggest that horizontal gene transfer has played a main role in shaping the haemolysin gene baggage in this pathogen.