Binary and mixed population biofilms: Time-lapse image analysis and disinfection with biocides

Simultaneous binary population biofilm formation by a bacterium and filamentous fungus was demonstrated by time-lapse image analysis in a flow cell system. The accumulation of attached bacterial cells followed an S-shaped graph similar to batch culture bacterial growth, with continual attachment, detachment, rotation, and movement of bacteria over the surface. An extensive hyphal network formed on the surface of the flow cell, protruding into the bulk flow, which subsequently detached. Multiple species mixed fungal-bacterial model biofilms were tested for isothiazolone biocide susceptibility. Biofilms were less susceptible to biocide treatment than planktonic cells of the same organisms. Mixed species biofilms, particularly for the bacterial species, offered greater protection against the action of the biocide compared to single species biofilms. Microbial loss as a result of biocide activity was shown by reduced cell surface coverage in electron micrographs.