Pseudomonadaceae increased the tolerance of Listeria monocytogenes to sanitizers in multi-species biofilms

Abstract

The persistence of the foodborne pathogen Listeria monocytogenes in food processing facilities may be facilitated by the formation of multi-species biofilms by environmental microbiota. This study aimed to determine whether multi-species biofilm formation results in an increased tolerance of L. monocytogenes in biofilms to the sanitizers benzalkonium chloride (BAC) and peroxyacetic acid (PAA) at concentrations commonly used in food processing facilities. Biofilms composed of microbiota previously shown to co-occur with L. monocytogenes in tree fruit packing facilities (i.e., Pseudomonadaceae, Xanthomonadaceae, Flavobacteriaceae, and Microbacteriaceae) were formed with L. monocytogenes in single- and multi-family assemblages. Multi-family biofilms were exposed to 250 or 500 ppm of PAA, or 200 ppm of BAC to determine the die-off kinetics of L. monocytogenes. Furthermore, the ability of a commercial biofilm remover to disrupt biofilms and inhibit bacteria in the formed single- and multi-family assemblage biofilms was assessed. The die-off kinetics of total bacteria and L. monocytogenes in biofilm assemblages throughout the exposure to a sanitizer was determined using the aerobic plate count and the most probable number methods, respectively. Biofilm assemblages that included Pseudomonadaceae resulted in an increased tolerance of L. monocytogenes to BAC and PAA compared to biofilm assemblages without Pseudomonadaceae. Further, the use of the biofilm remover significantly disrupted biofilms and reduced the concentration of L. monocytogenes in single- and multi-family biofilms by 5 or more logarithmic units. These findings highlight the need to improve the control of biofilm-forming microbiota in food processing facilities to mitigate the persistence of L. monocytogenes.

Keywords: Antimicrobial tolerance; Benzalkonium chloride; Biofilm remover; Environmental microbiota; Flavobacteriaceae, biofilms; Listeria monocytogenes; Microbacteriaceae; Peroxyacetic acid; Pseudomonadaceae; Sanitizers; Xanthomonadaceae.