Biocide tolerant Listeria monocytogenes isolates from German food production plants do not show cross-resistance to clinically relevant antibiotics.

Contamination of food during processing is recognized as main transmission route of Listeria (L.) monocytogenes. To prevent microbial contamination, biocides are widely applied as disinfectants in food processing plants. However, there are concerns about the development of antimicrobial resistance in foodborne pathogens due to widespread biocide usage. In our study, 93 L. monocytogenes isolates from German food production facilities were i) tested for biocide and antibiotic susceptibility using broth microdilution assays, ii) analyzed for links between reduced biocide susceptibility and antibiotic resistance, and iii) characterized by whole-genome sequencing including the detection of genes encoding for biocide tolerance, antibiotic resistance, and other virulence factors. Fifteen L. monocytogenes isolates were tolerant to benzalkonium chloride (BAC) and genes conferring BAC tolerance were found in 13 of them. Antibiotic resistance was not associated with biocide tolerance. BAC tolerant isolates were assigned to 6 MLST clonal complexes and most of them harbored internalin A pseudogenes with premature stop codons or deletions (n=9). Our study demonstrated a high genetic diversity among the investigated isolates including genotypes that are frequently involved in human infections. Although in vitro adaptation studies to biocides have raised concerns about increasing cross-resistance to antibiotics, our results do not provide evidence for this phenomenon in field isolates. Importance Foodborne pathogens such as L. monocytogenes can persist in food production environments for a long time causing perennial outbreaks. Hence, bacterial pathogens are able to survive cleaning and disinfection procedures. Accordingly, they may be repeatedly exposed to sublethal concentrations of disinfectants, which might result in bacterial adaptation to these biocides. Furthermore antibiotic co- and cross-resistance is known to evolve under biocide selection pressure in vitro. Hence, antimicrobial tolerance seems to play a crucial role in resilience and persistence of foodborne pathogens in the food chain and might reduce therapeutic options in infectious diseases.