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Recently, considerable attention has been paid to the biologically active of fruit and vegetable processing wastes. In this study, antibacterial properties and major bioactive constituents of Chinese water chestnut peels (CWCP) extracts and fractions were evaluated. The data displayed that ethyl acetate fractions (EF) from CWCP showed greater antibacterial activity than n-butanol fractions, methanol extracts and water fractions against three common foodborne pathogenic bacteria ( Staphylococcus aureus, Escherichia coli and Listeria monocytogenes) assessed by the inhibition zone, minimal inhibition concentration and minimal bactericidal concentration values. In accordance with disc diffusion assay, S. aureus was more susceptible to EF than the other two pathogens. Furthermore, action-mode studies indicated that EF exhibited significant bactericidal activity against S. aureus by degrading and disrupting the cell wall and cytoplasmic membrane, ultimately the leakage of cell contents led to cell death. Finally, high-performance liquid chromatography coupled with photodiode array and electrospray ionization mass spectrometry was used for characterization of bioactive constituents in EF, and ten flavonoids aglycons were identified or tentatively identified. The results suggest that the extracts and fractions from CWCP could be potentially used as a possible food supplement to improve food safety by the control or elimination of foodborne pathogenic bacteria. [ABSTRACT FROM AUTHOR]