In 2023, Rana dybowskii exhibiting characteristic skin ulcers were found on a farm in northeastern China. Subsequently, two dominant bacteria, Aeromonas hydrophila Rd001 and Acinetobacter johnsonii Rd002, were isolated from naturally infected R. dybowskii . Experimental infection confirmed that Rd001 was the primary pathogen responsible for the disease in R. dybowskii , with a mean lethal dose (LD 50 ) of 6.25 × 10 2 CFU/g. The virulence genotype of Rd001 was identified as ser +/ aha +/ lip +/ nuc +/ hlyA +/ aer +/ alt +/ ast +/ act +. Antimicrobial susceptibility testing indicated that Rd001 was sensitive to enrofloxacin, flumequine, and neomycin. MLST analysis showed that Rd001 belonged to a new sequence type of A. hydrophila , named ST2378. This study offered the first comprehensive investigation into the pathogenicity, virulence genotypes, antimicrobial resistance, and genetic traits of A. hydrophila isolated from R. dybowskii , providing a theoretical foundation for preventing and controlling A. hydrophila infections.