Highlighting the limitations of static microplate biofilm assays for industrial biocide effectiveness compared to dynamic flow conditions.

ABSTRACT

The minimal inhibitory concentration of an antimicrobial required to inhibit the growth of planktonic populations (minimum inhibitory concentration [MIC]) remains the 'gold standard' even though biofilms are acknowledged to be recalcitrant to concentrations that greatly exceed the MIC. As a result, most studies focus on biofilm tolerance to high antimicrobial concentrations, whereas the effect of environmentally relevant sub-MIC on biofilms is neglected. The effect of the MIC and sub-MIC of an isothiazolinone biocide on a microbial community isolated from an industrial cooling system was assessed under static and flow conditions. The differential response of planktonic and sessile populations to these biocide concentrations was discerned by modifying the broth microdilution assay. However, the end-point analysis of biofilms cultivated in static microplates obscured the effect of sub-MIC and MIC on biofilms. A transition from batch to the continuous flow system revealed a more nuanced response of biofilms to these biocide concentrations, where biofilm-derived planktonic cell production was maintained despite an increase in the frequency and extent of biofilm sloughing. A holistic, 'best of both worlds' approach that combines the use of static and continuous flow systems is useful to investigate the potential for the development of persistent biofilms under conditions where exposure to sub-MIC and MIC may occur.