[Objective]The aim of this study was to investigate the disinfection efficacy of slightly acidic electrolyzed water (SAEW) and gain a better understanding of the antibacterial mode of action against Staphylococcus aureus.[Methods]The application of flow cytometry with double-staining method for a rapid and reliable assessment of cell viability and functionality were compared with traditional colony forming unit (CFU) culture method.The further analysis of sublethal and viable but nonculturable (VBNC) state of S.aureus was to elucidate possible induction of SAEW on S.aureus.Scanning and transmission electron microscopy were also used to obtain distinct images of external shape and internal structure changes of S.aureus.[Resultsl We observed that the germicidal efficiency decreased gradually with the increase of duration time,and 3.06 log CFU/mL elimination ofS.aureus was achieved after treated with 2 mg/L SAEW for 10 min.The processing time of 10 min in this study was long enough to avoid the production of sublethal and VBNC state of S.aureus.The destructive action of SAEW was a short-time sterilization mode and had multiple target sites,including the cell wall,the cytoplasmic membrane,and the nucleic acids.[Conclusion]The results indicated that SAEW was an effective disinfectant for environmental remediation and food preservation even at low available chlorine concentration.