Specific phenotypic, genomic, and fitness evolutionary trajectories toward streptomycin resistance induced by pesticide co-stressors in Escherichia coli

To explore how co-occurring non-antibiotic environmental stressors affect evolutionary trajectories toward antibiotic resistance, we exposed susceptible Escherichia coli K-12 populations to environmentally relevant levels of pesticides and streptomycin for 500 generations. The coexposure substantially changed the phenotypic, genotypic, and fitness evolutionary trajectories, resulting in much stronger streptomycin resistance (>15-fold increase) of the populations. Antibiotic target modification mutations in rpsL and rsmG, which emerged and dominated at late stages of evolution, conferred the strong resistance even with less than 1% abundance, while the off-target mutations in nuoG, nuoL, glnE, and yaiW dominated at early stages only led to mild resistance (2.5-6-fold increase). Moreover, the strongly resistant mutants exhibited lower fitness costs even without the selective pressure and had lower minimal selection concentrations than the mildly resistant ones. Removal of the selective pressure did not reverse the strong resistance of coexposed populations at a later evolutionary stage. The findings suggest higher risks of the selection and propagation of strong antibiotic resistance in environments potentially impacted by antibiotics and pesticides.

Automated summary

The co-exposure of E. coli populations to pesticides and streptomycin in environmentally relevant levels significantly impacted phenotypic, genotypic, and fitness evolutionary trajectories, leading to stronger streptomycin resistance. Mutations in antibiotic target genes emerged in late stages of evolution conferred strong resistance even at low abundance, while off-target mutations dominated in early stages but only led to mild resistance. High resistant mutants showed lower fitness costs and minimal selection concentrations compared to mildly resistant mutants, and removal of selective pressure did not reverse strong resistance at later evolutionary stages.