O157:H7 strains associated with several recent (2017-2020) multi-state outbreaks linked to leafy green vegetables have been characterized as "reoccurring, emerging, and persistent" (REP). Our recent unpublished work demonstrated that the REP strains had significantly enhanced potential for biofilm formation. In this study, comparative genomic analyses were conducted for a better understanding of the mechanisms behind the enhanced biofilm formation, and thereby potentially increased environmental fitness, by the REP strains. Phylogenetically, the recent outbreak strains formed two distinct clusters represented by REPEXH01 and REPEXH02. Compared with EDL933 and other previous outbreak reference strains, the REP strains (clustering with REPEXH02) exhibiting strong biofilm formation were found to have acquired two genes encoding proteins of unknown functions (hypothetical proteins) and lost certain prophage-related genes. In addition, several single nucleotide polymorphisms in genes related to biofilm formation were identified.
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