Lactic acid bacteria were isolated from various food samples and evaluated for hydrogen peroxide (H2O2) production. Cells suspended in 0.5% (wt/vol) glucose plus 0.5% (wt/vol) lactate (pH 7.0) were incubated for 5 h at 37 degreesC under aeration. Among 193 strains, 27 strains accumulated 201-300 ppm H2O2, and 4 strains accumulated more than 301 ppm H2O2 in the cell suspensions. Among the 9 high-level H2O2-producing strains, 8 strains were identified as Lactococcus lactis subsp. lactis. The cell-free filtrate from Lc. lactis subsp. lactis AI 62, which contained approximately 350 ppm H2O2, was evaluated for antimicrobial activity against Enterococcus faecalis, Ent. faecium, enterotoxigenic Escherichia coli, Listeria ivanovii, Staphylococcus aureus, Yersinia enterocolitica, and Aeromonas hydrophila. After 1 h incubation at 30 degreesC in the cell-free filtrate, the initial viable cell counts of the target bacteria (5.53-6.00 log cfu/mL) were reduced by 0.12-5.00 log units, except in the case of enterococci. The sensitivity varied with the bacterial species and pH. The enterococci were resistant to the treatment. Our results show that H2O2 accumulated by lactic acid bacteria in a cell suspension is very effective in reducing the viable cell count of food-borne pathogens.