Emergence and characteristics of multidrug-resistant Salmonella enterica subspecies enterica serovar Infantis harboring the pESI plasmid in chicken slaughterhouses in South Korea

Abstract

Salmonella enterica subsp. enterica serovar Infantis (S. Infantis), a major cause of human salmonellosis, is commonly associated with transmission via contaminated chicken meat. This study, as part of the national Salmonella monitoring program, assessed the prevalence of S. enterica, including S. Infantis, in chicken slaughterhouses across South Korea from 2014-2022. The presence of a megaplasmid, known as plasmid of emerging S. Infantis (pESI), was confirmed. This confirmation was based on multidrug-resistant and third-generation cephalosporin-resistant S. Infantis isolates using whole-genome sequencing. Phenotypic and genotypic characterization involved antimicrobial susceptibility tests, pulsed-field gel electrophoresis, polymerase chain reaction to screen for pESI plasmids, plasmid profiling, and conjugation assays. S. Infantis was identified in 9.3% of Salmonella-positive samples in 2014, undetected from 2015-2020, but re-emerged as the predominant serovar in 2021 (54.7%) and 2022 (75.5%). The isolates in 2014 were antibiotic susceptible, whereas most isolates from 2021 to 2022 exhibited multidrug-resistance, including resistance to third-generation cephalosporins. All isolates were sequence type 32 (ST32), with core genome multilocus sequence typing demonstrating pESI plasmid-based clustering. The pESI⁺ isolates harbored genes, such as aadA1, dfrA14, sul1, and tetA, and three multidrug-resistant pESI⁺ isolates harbored bla_CTX-M-65. The plasmids were genetically similar to those observed in S. Infantis from broilers, chicken meat, and human clinical samples across various countries. This study highlights the spread of multidrug-resistant S. Infantis harboring the pESI plasmid with bla_CTX-M-65 during early chicken production in South Korea. Continuous monitoring and control of resistant S. Infantis throughout the food chain is essential to inform public health initiatives.IMPORTANCEThis study highlights the critical emergence of multidrug-resistant (MDR) Salmonella enterica serovar Infantis (S. Infantis) in South Korea's chicken slaughterhouses, driven by the acquisition of the pESI megaplasmid harboring the extended-spectrum beta-lactamase (ESBL) determinant bla_CTX-M-65. Using whole-genome sequencing and comprehensive phenotypic-genotypic analyses, the findings reveal that pESI⁺ isolates in South Korea are genetically similar to strains from broilers, chicken meat, and human clinical cases worldwide. This underscores the transboundary nature of S. Infantis and its potential as a global public health threat.

Keywords: ST32; Salmonella Infantis; blaCTX-M-65; chicken slaughterhouses; multidrug resistance; pESI.

Fig 1

Prevalence of Salmonella enterica and serovar Infantis in chicken slaughterhouses in South Korea during 2014–2022.

Fig 2

PFGE analysis and antimicrobial resistance patterns of 95 S. Infantis isolates from chicken slaughterhouses in South Korea (2014–2022). CB, Chungbuk; CN, Chungnam; GB, Gyeongbuk; GN, Gyeongnam; JB, Jeonbuk; JN, Jeonnam; AMP, ampicillin; AUG, amoxicillin-clavulanic acid; AXO, ceftriaxone; AZI, azithromycin; CHL, chloramphenicol; CIP, ciprofloxacin; FIS, sulfisoxazole; FOX, cefoxitin; GEN, gentamicin; NAL, nalidixic acid; STR, streptomycin; SXT, trimethoprim-sulfamethoxazole; TET, tetracycline; XNL, ceftiofur.

Fig 3

cgMLST tree of 7 Salmonella Infantis strains isolated from chicken slaughterhouses in this study and 94 S. Infantis isolates obtained from broiler, chicken meat, and humans in 15 countries registered in public databases (NCBI or ENA; data collected on 20 December 2023). Circular representation of the phylogeny was visualized using iTOL (http://itol.embl.de/) ignoring branch length. The tree includes information on the isolates, such as isolation year, source, and presence of pESI and beta-lactam resistance genes.

Fig 4

Pangenome analysis of four South Korean pESI plasmids (CS72S01, CS72S03, CS78S02, and CS81S04) compared with the original Israeli pESI and other pESI-like plasmids. (A) Pangenome analysis; (B) resistance region 1; (C) resistance region 2.