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Bactericidal effects of Polygonum cuspidatum extract on Bacillus cereus were detected through measurement ofintracellular- and extracellular adenosine triphosphate (ATP), and analyses with transmission electron microscope (TEM) andscanning electron microscope (SEM). TEM analysis revealed B. cereus treated with P. cuspidatum volatile extracts obtained bysimultaneous steam distillation and solvent extraction (SDE) method showed intracellular shape change. SEM analysisrevealed several holes on cell wall of bacteria treated with SDE extracts of P. cuspidatum. Intracellular ATP concentrations ofthe cells treated with P. cuspidatum or 2-hexenal decreased to 0.272 and 0.232 μM from 0.618 μM, respectively. ExtracellularATP concentrations of control, and bacteria treated with P. cuspidatum extracts and 2-hexenal were 0.062, 0.301, and 0.326μM, showing increases of 4.9- and 5.3-fold, respectively. Results suggest bactericidal effects of P. cuspidatum SDE extractsare due to change in intracellular shape or permeability caused by damage to cell membrane.