Abstract

Contamination of food during processing is recognized as a main transmission route of Listeria 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 coding 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 multilocus sequence type (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 coresistance and cross-resistance are known to evolve under biocide selection pressure in vitro Hence, antimicrobial tolerance seems to play a crucial role in the resilience and persistence of foodborne pathogens in the food chain and might reduce therapeutic options in infectious diseases.

Highlights

  • IMPORTANCE Foodborne pathogens such as L. monocytogenes can persist in food production environments for a long time, causing perennial outbreaks

  • Our study revealed a high genetic diversity among L. monocytogenes isolates from technical equipment and surfaces of German food production facilities

  • The detection of genotypes that are frequently involved in human listeriosis highlights the importance of contaminated food production environments as transmission routes for virulent L. monocytogenes

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Summary

Introduction

IMPORTANCE Foodborne pathogens such as L. monocytogenes can persist in food production environments for a long time, causing perennial outbreaks. Increased tolerance against antimicrobial stress triggered by the application of disinfectants may be an important factor for the persistence of L. monocytogenes in food production environments [20, 21]. The aim of our study was to examine the biocide susceptibilities of L. monocytogenes isolates from food production plants in Germany and to look for potential relationships between biocide tolerance and antibiotic resistance. We tested the susceptibilities to six antimicrobial biocides frequently used in the food industry and to antibiotics relevant for human listeriosis therapy. We analyzed the genetic diversity of the L. monocytogenes strains under study and investigated the prevalence of putative biocide tolerance and antibiotic resistance genes as well as virulence genes

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