| Summary: | <i>Listeria monocytogenes</i> is one of the main foodborne pathogens. The formation of biofilms by <i>L. monocytogenes</i> contributes to its resistance to disinfectants, which represents a serious risk for food production plants. The aim of this study was to compare the effect of sub-inhibitory concentrations of benzalkonium chloride (BAC) (1.25 or 2.5 mg/L) on biofilm production and on biofilm reduction after exposure to an inhibitory concentration of BAC (1280 mg/L) in two isogenic <i>L. monocytogenes</i> strains: the BAC-sensitive wild-type strain S2-1 and its BAC-resistant mutant derivative S2<sup>BAC</sup>, which presented a multidrug resistance phenotype. The biofilm-forming ability of the strains under different BAC concentrations was evaluated by the resazurin method using polystyrene microplates. The biofilm reduction after BAC exposure was evaluated by using stainless steel coupons (SSCs). When the resazurin method was used, S2<sup>BAC</sup> produced significantly more biofilm in the presence of a sub-inhibitory concentration of BAC compared to that in the culture medium without BAC (<i>p</i> < 0.05). When the SSC method was used, the presence of sub-inhibitory concentrations of BAC resulted in a higher resistance of the biofilm for S2<sup>BAC</sup> compared to that in the culture medium without BAC (<i>p</i> < 0.05). This was not observed with the sensitive S2-1 strain. These results suggest that biofilm behavior depends on the strain and sub-inhibitory concentrations of disinfectants and may explain the ability of certain isolates to persist in niches of food processing plants.
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