Large-scale phenotypic and genomic characterization ofListeria monocytogenessusceptibility to quaternary ammonium compounds
Authors: Ivanova Mirena , Kragh Martin Laage , Szarvas Judit , Tosun Elif Seyda , Holmud Natacha Friis , Gmeiner Alexander , Amar Corinne , Guldimann Claudia , Huynh TuAnh N. , Karpíšková Renáta , García Carmen Rota , Gomez Diego , Aboagye Eurydice , Etter Andrea , Centorame Patrizia , Torresi Marina , De Angelis Maria Elisabetta , Pomilio Francesco , Okholm Anders Hauge , Xiao Yinghua , Kleta Sylvia , Lueth Stefanie , Pietzka Ariane , Kovacevic Jovana , Pagotto Franco , Rychli Kathrin , Zdovc Irena , Papić Bojan , Heir Even , Langsrud Solveig , Møretrø Trond , Stephan Roger , Brown Phillip , Kathariou Sophia , Tasara Taurai , Aarestrup Frank , Njage Patrick Murigu Kamau , Fagerlund Annette , Hansen Lisbeth Truelstrup , Leekitcharoenphon Pimlapas 
Published: September 2023

ABSTRACTListeria monocytogenesis a significant concern for the food industry due to its ability to persist in the food processing environment. Decreased susceptibility to disinfectants is one of the factors that contribute to the persistence ofL. monocytogenes. The objective of this study was to explore the diversity ofL. monocytogenessusceptibility to quaternary ammonium compounds (QACs) using 1,671L. monocytogenesisolates. This was used to determine the phenotype-genotype concordance and characterize genomes of the QAC sensitive and tolerant isolates for stress resistance, virulence and plasmid replicon genes. Distribution of QAC tolerance genes among 37,897 publicly availableL. monocytogenesgenomes were also examined. The minimum inhibitory concentration to QACs was determined by the broth microdilution method and non-sequenced isolates (n=1,244) were whole genome sequenced. Genotype-phenotype concordance was 99% for benzalkonium chloride, DDAC and a commercial QAC based sanitizer. Prevalence of QAC tolerance genes was 23% and 28% in ourL. monocytogenescollection and in the global dataset, respectively.qacHwas the most prevalent gene in our collection (61%), with 19% prevalence in the global dataset. Notably,bcrABCwas most common (72%) globally, while 25% in our collection. Prevalence ofemrCandemrEwas comparable in both datasets, 7% and 2%, respectively. Replicon genes, indicative of plasmid harborage, were detected in 44% of the isolates and associated with the QAC tolerant phenotype. The presented analysis is based on the biggestL. monocytogenescollection in diversity and quantity for characterization of theL. monocytogenesQAC tolerance at both phenotypic and genomic levels.IMPORTANCEContamination ofListeria monocytogeneswithin the food processing environment is of concern to the food industry due to challenges in eradicating the pathogen once it becomes persistent in the environment. Genetic markers associated with increased tolerance to disinfectants have been identified, which alongside factors favor the persistence ofL. monocytogenesin the production environment. By employing a comprehensive large-scale phenotypic testing and genomic analysis our study significantly enhances the understanding of the prevalence of quaternary ammonium compound (QAC) tolerantL. monocytogenesand the genetic determinants associated with the increased tolerance. Furthermore, we report on the prevalence of QAC tolerance genes among 37,897 publicly availableL. monocytogenessequences and their distribution within clonal complexes, isolation sources and geographical locations. As the propagation of QAC tolerance showed not be evenly distributed globally this highlights that understanding the development ofL. monocytogenesdisinfectant tolerance can be monitored using publicly available WGS data.