A multidrug-resistant Salmonella enterica serotype Typhimurium DT104 lineage circulating among humans and cattle in the United States lost the ability to produce pertussis-like toxin ArtAB in close temporal proximity to the global DT104 epidemic

Abstract

Salmonella enterica subspecies enterica serotype Typhimurium definitive type 104 (DT104) can infect both humans and animals and is often multidrug-resistant (MDR). Previous studies have indicated that, unlike most S. Typhimurium, the overwhelming majority of DT104 strains produce pertussis-like toxin ArtAB via prophage-encoded genes artAB. However, DT104 that lack artAB have been described on occasion. Here, we identify a MDR DT104 complex lineage circulating among humans and cattle in the United States, which lacks artAB (i.e., the "U.S. artAB-negative major clade"; n = 42 genomes). Unlike most other bovine- and human-associated DT104 complex strains from the U.S. (n = 230 total genomes), which harbor artAB on prophage Gifsy-1 (n = 177), members of the U.S. artAB-negative major clade lack Gifsy-1, as well as anti-inflammatory effector gogB. The U.S. artAB-negative major clade encompasses human- and cattle-associated strains isolated from [≥]11 U.S. states over a twenty-year period. The clade was predicted to have lost artAB, Gifsy-1, and gogB circa 1985-1987 (95% highest posterior density interval 1979.0-1992.1). When compared to DT104 genomes from other world regions (n = 752 total genomes), several additional, sporadic artAB, Gifsy-1, and/or gogB loss events among clades encompassing [≤]5 genomes were observed. Using phenotypic assays that simulate conditions encountered during human and/or bovine digestion, members of the U.S. artAB-negative major clade did not differ from closely related Gifsy-1/artAB/gogB-harboring U.S. DT104 complex strains (ANOVA raw P-value > 0.05); thus, future research is needed to elucidate the roles that artAB, gogB, and Gifsy-1 play in DT104 virulence in humans and animals. Impact StatementMulti-drug resistant (MDR) Salmonella enterica serotype Typhimurium definitive type 104 (DT104) was responsible for a global epidemic among humans and animals throughout the 1990s and continues to circulate worldwide. Previous studies have indicated that the vast majority of DT104 produce pertussis-like toxin ArtAB via prophage-encoded artAB. Here, we identify a DT104 complex lineage that has been circulating among cattle and humans across [≥]11 U.S. states for over twenty years, which lacks the ability to produce ArtAB (i.e., the "U.S. artAB-negative major clade"). The common ancestor of all U.S. artAB-negative major clade members lost the ability to produce ArtAB in the 1980s; however, the reason for this loss-of-function event within this well-established pathogen remains unclear. The role that ArtAB plays in DT104 virulence remains elusive, and phenotypic assays conducted here indicate that members of the U.S. artAB-negative major clade do not have a significant advantage or disadvantage relative to closely related, Gifsy-1/artAB/gogB-harboring U.S. DT104 complex strains when exposed to stressors encountered during human and/or bovine digestion in vitro. However, ArtAB heterogeneity within the DT104 complex suggests clade-specific selection for or against maintenance of ArtAB. Thus, future studies querying the virulence characteristics of the U.S. artAB-negative major clade are needed. Data SummarySupplementary Data is available under DOI 10.5281/zenodo.7688792, with URL https://doi.org/10.5281/zenodo.7688792.