Human infection by Legionella pneumophila (Lpn) only occurs via contaminated water from man-made sources, and eradication of these bacteria from man-made water systems is complicated by biofilm colonization. Using a continuously fed biofilm reactor model, we grew a biofilm consortium from potable water that was able to prolong recovery of Lpn CFU from biofilms. This effect was recreated using a subset of those species in a simplified consortium composed of eight bacterial isolates from the first biofilm reactor. In the reactor with the eight-species consortium, Lpn biofilm CFU was relatively stable over a 12-day trial. An isolate of Acidovorax from the consortium was, as a single species biofilm, able to promote Lpn surface attachment. Other isolates from the Pelomonas genus grew as equally robust biofilms alone, but did not promote surface attachment of Lpn. This attachment was disrupted by cationic polysaccharides and loss of the Lpn Lcl collagen-like adhesin protein. This work demonstrates that, while Lpn was fairly incompetent at attachment to surfaces to form a biofilm alone, pre-existing biofilms allowed attachment of Lpn as secondary colonizers. In addition, we demonstrate that initial attachment of Lpn to Acidovorax biofilms is likely via the Lcl-adhesin protein.

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