Co-occurrence of biofilm formation, acid tolerance, and antibiotic resistance in environmental Escherichia coli associated with lettuce.

backgroundEnvironmental niches represent important reservoirs of Escherichia coli with stress-adaptation traits that support persistence outside the host. Contaminated irrigation water and soils can facilitate transfer to fresh produce, where bacterial survival may reduce the effectiveness of downstream control measures. This study investigated the co-occurrence of biofilm formation, acid tolerance, and antibiotic resistance (AR) in environmental E. coli and their contribution to persistence along the farm-to-produce continuum.

resultsEighteen E. coli isolates recovered from irrigation water, soil, and lettuce were characterized using phenotypic assays and genome-based analyses. Most isolates remained susceptible to the majority of tested antibiotics, with multidrug resistance observed in only 11.1% of isolates. In contrast, moderate-to-strong biofilm formation was widespread (83.3%), and several isolates exhibited reduced susceptibility to acetic acid at concentrations relevant to household washing practices. Genotypic screening revealed a broad distribution of adhesion, iron acquisition, biofilm-associated, and plasmid-borne resistance determinants, indicating substantial functional diversity. Significant positive associations were observed between acid tolerance, biofilm formation, and antibiotic resistance, suggesting co-occurrence of stress-adaptation phenotypes rather than definitive evolutionary convergence. While antibiotic resistance phenotypes showed strong concordance with corresponding resistance genes, biofilm formation and acid tolerance were not associated with specific genetic determinants, supporting a multifactorial basis of these traits.

conclusionsThese findings demonstrate that environmental E. coli can combine multiple stress-adaptation mechanisms that enhance persistence across agricultural and food-associated environments, even in the absence of high-risk resistance profiles. The observed co-occurrence of phenotypic traits highlights the potential for co-selection under environmental pressures and underscores the limitations of relying solely on downstream decontamination strategies. Effective risk mitigation requires integrated, preventive approaches targeting pre-harvest contamination and environmental reservoirs.