Whole-genome sequencing-based characterization of Listeria monocytogenes isolated from cattle and pig slaughterhouses.

Listeria monocytogenes is a foodborne pathogen that causes human listeriosis and may be transmitted to humans via the food chain, beginning at slaughter and extending through food production and consumption. In this study, we performed whole-genome sequencing (WGS) analysis to determine the genetic characteristics of L. monocytogenes from the carcasses and environments of cattle and pig slaughterhouses in Korea. In total, 50 L. monocytogenes isolates were collected from 46 cattle and 47 pig slaughterhouses nationwide from 2014 to 2022. They were classified into two lineages, 12 sublineages, 12 sequence types, 11 clonal complexes (CCs), and 15 core-genome multilocus sequence typing types. L. monocytogenes isolates were divided into two lineage: lineage I (serotypes 1/2b and 4b) and lineage II (serotypes 1/2a and 1/2c). The most frequent CCs were CC9 (46.0 %), followed by CC224 (16.0 %) and CC155 (14.0 %). Although all isolates exhibited highly conserved LIPI-1, 20.0 % and 2.0 % contained LIPI-3 or LIPI-4, respectively. Moreover, 96.0 % of the isolates had full-length inlA. Interestingly, 21 of the 23 CC9 isolates contained mutations in inlA resulting from premature stop codon (PMSC). The mdrL and Listeria genomic island-2 (LGI-2) were identified in all L. monocytogenes isolates, whereas LGI-3 was identified in 32.0 % of the isolates. The L. monocytogenes isolates contained various antimicrobial resistance genes, moreover, the plasmid-borne resistance genes tetM and mprF were also identified in 34.0 % and 100 % of the isolates, respectively. Twenty-four isolates (48.0 %) harbored one or two plasmids (pLM33, DOp1, pLGUG1, and pLM5578), and 29 isolates (58.0 %) harbored at least one insertion sequence, composite transposon, and integrative conjugative element. Four isolates showed two CRISPR-Cas types IB and II-A. In addition, phage sequences associated with the spacer constituting the CRISPR array were identified in 26 Listeria phages from 14 L. monocytogenes isolates. The genetic composition of L. monocytogenes was conserved in a collinearity relationship between each of the five L. monocytogenes isolates from the cattle and pig slaughterhouses. These findings suggest that L. monocytogenes isolated from cattle and pig slaughterhouses have the ability to cause human disease and exhibit virulent characteristics.