108 microorganism types (79 types of fungi and 29 types of bacteria) were isolated from 25 automobiles generating bad odor when the air conditioner was turned on, and 43 types of fungi and 23 types of bacteria were identified. The analysis of condensate generated by the air conditioners in the automobiles indicated pH 6.4~7.1, 12.5~34.2 mg/L carbon source, 0.9~18.6 mg/L nitrogen source, 0.5~27.8 mg/L ion contents, and 0.1~7.7 mg/L mineral contents. The biofilms were formed by the mesophiles under the summer temperature/humidity condition (26 oC, 70% R.H.), and they were regenerated when the environmental factors (nutritional contents and temperature/humidity) were appropriate even after they were artificially removed. Although the antimicrobials removed 99.9% of planktonic cells within 15 minutes, they were not effective in removing biofilm. Up to 1,950 ppmv of ethanol was observed in the automobile treated with the antimicrobials. Although the figure is lower than the acute toxicity level when inhaled by humans, the health safety of the chemical substances used in the antimicrobials needs to be reviewed.