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This study investigated the influence of N-chlorosuccinimide (NCS) pretreatment on the antimicrobial effect of benzalkonium chloride (BZC, representative of QACs) against biofilm bacteria and its mechanisms. Results show that 0.04 − 0.07 mmol/L NCS pretreatment significantly increased the antimicrobial efficacy of 0.03 mmol/L BZC on biofilm cells by 30% − 70%. The main mechanisms involved membrane permeability, oxidative damage, and metabolic disorder. More precisely, NCS pretreatment increased the permeability of bacteria and reduced the activity of the electron transport system (ETS) and dehydrogenase (DHA). At the same time, the oxidative damage of both endogenous and exogenous ROS and the disorder of the antioxidant enzymes (superoxide dismutase and catalase) further improved their combined antibacterial ability. Moreover, NCS pretreatment greatly reduced the resistance of biofilm  Pseudomonas aeruginosa  to BZC. The findings of the study provide a new method to effectively enhance the antimicrobial efficiency of quaternary ammonium cationic surfactants (e.g., BZC) and reduce bacterial resistance, as well as a scientific guidance for the development of new antimicrobial products.

N-chlorosuccinimide enhancing the antimicrobial effect of benzalkonium chloride on biofilm Pseudomonas aeruginosa and its interaction mechanisms