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The susceptibility breakpoints are crucial for prudent use of antimicrobials.This study has firstly developed a susceptibility breakpoint (MIC≤0.25 μg/mL) of enrofloxacin against swine Salmenolla spp based on wild-type cutoff (COWT) and pharmacokinetic-pharmadynamic (PK-PD) cutoff (COPD), consequently providing a criteria for susceptibility testing and clinical usage of enrofloxacin.