Journal of Antimicrobial Chemotherapy · 2022 · 14 citations · 8 references
Life SciencesGeneticsMicrobiologyMicrobial EvolutionMicrobial GeneticsMicroevolutionBacterial GeneticsHorizontal Gene TransferPlasmid BiologyPlasmid GeneticsPlasmid-mediated ResistanceScreening Of MutantsAntibiotic Resistance GenesChromosomal MutationsChromosomal Resistance MutationsHigh-throughput Screening AssaysPlasmid CuringPlasmid EvolutionPlasmid Fitness CostRelative Fitness
OBJECTIVES: To investigate the fitness effects of large blaCTX-M-15-harbouring F2:A1:B- plasmids on their native Escherichia coli ST131 H30Rx hosts. METHODS: We selected five E. coli ST131 H30Rx isolates of diverse origin, each carrying an F2:A1:B- plasmid with the blaCTX-M-15 gene. The plasmid was eliminated from each isolate by displacement using an incompatible curing plasmid, pMDP5_cureEC958. WGS was performed to obtain complete chromosome and plasmid sequences of original isolates and to detect chromosomal mutations in 'cured' clones. High-throughput competition assays were conducted to determine the relative fitness of cured clones compared with the corresponding original isolates. RESULTS: We were able to successfully eliminate the F2:A1:B- plasmids from all five original isolates using pMDP5_cureEC958. The F2:A1:B- plasmids produced non-significant fitness effects in three isolates and moderate reductions in relative fitness (3%-4%) in the two remaining isolates. CONCLUSIONS: We conclude that F2:A1:B- plasmids pose low fitness costs in their E. coli ST131 H30Rx hosts. This plasmid-host fitness compatibility is likely to promote the maintenance of antibiotic resistance in this clinically important E. coli lineage.
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An efficient stress-free strategy to displace stable bacterial plasmids
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