Pathogenome and Plasmid-Borne Antimicrobial Resistance Phenotypes in a Multidrug-Resistant O111:H8 Shiga...

Plasmids contribute to virulence and antimicrobial resistance in Shiga toxin-producing Escherichia coli (STEC). Here, the mobility, gene content, and evolutionary context of four plasmids carried by an O111:H8 STEC strain designated UTAK-22: pUTAK-22.1-MDR, pUTAK-22.2-MDR, pUTAK-22.3-P1, and pUTAK-22.4-pO111 were characterized. Conjugation experiments, phenotype-based selective readouts, and comparative genomics together showed that pUTAK-22.1-MDR is self-transmissible, whereas pUTAK-22.2-MDR and pUTAK-22.4-pO111 are most consistent with mobilization in trans, making use of the strain's native helper plasmid background, while pUTAK-22.3-P1 represents a conjugation-deficient IncY phage-derived replicon. Genome annotation and comparative analyses further highlighted the structural diversity and mosaic composition of these plasmids, including P1-like phage remnants, virulence loci, and distinct antimicrobial resistance modules. Phenotypic profiling of the native wild-type plasmid complement and selected transconjugants in the recipient E. coli strain WG5 further showed that the presence of co-resident plasmids and redundant resistance determinants results in dosage-dependent streptomycin tolerance. Together, these findings expand our understanding of the mobility landscape, evolutionary dynamics, and resistance potential of STEC plasmids, while underscoring the importance of interpreting resistance phenotypes in the context of a strain's natural plasmid composition.