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[Objective]In order to carriy on effective medication and immune prevention and control,pathogen and etiology of a Rana spinosa David disease in a Rana spinosa David breeding company in Zhejiang was researched from 2009 to 2010. [Methods]Conventional bacteria separation and purification technology was used to purify pathogenic bacteria,regression infection and toxicity test was used to determine its virulence,physiological and biochemical and molecular biology identification was carried out to the the pathogen,k- b paper method was adopted for the susceptibility test,normal paraffin wax flaking technology was used for pathology observations,finally,the inactivation conditions of pathogenic bacteria was explored. [Results]The pathogenic bacteria separated from sick Rana spinosa David body had Median Lethal Concentration（ LC 50） of 5.62 ×105 cfu / ml; the strain was identified as Aeromonas hydrophila by ATB bacteria identification instrument and 16 s rRNA sequence analysis（ GenBank login number： HQ322682）; histopathology observation results showed that the disease had main symptoms of the liver and kidney damage,inflammatory cells increase. Susceptibility test showed that the strain was highly sensitive to gentamicin,norfloxacin,florfenicol,enrofloxacin etc. [Conclusions]1% formaldehyde could well inactivate the pathogenic bacteria at 60 ℃ for 24 h,which would provide technical reference for whole bacteria inactivated vaccine preparation.