Evaluation of the bactericidal activity of plasma-activated NaCl solution (PAN) against foodborne pathogens: Inactivation mechanism and application to mackerel

The objective of this study was to assess the bactericidal effect of plasma-activated NaCl solution (PAN) against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes and apply PAN as a brine salting solution for mackerel. To enhance the bactericidal effect of plasma-activated water (PAW), NaCl solutions (0, 3.5, 7, and 10%) were treated with plasma for 20 and 40 min to generate PAN. Plasma-activated glycerol solution (PAG) was also included to evaluate the influence of water activity on plasma activation and its effect on microbial activity. Physicochemical analysis revealed that elevating the NaCl concentration of PAN led to a decrease in pH, an increase in oxidation–reduction potential, and higher levels of reactive species such as H₂O₂ and HOCl. PAN showed greater antibacterial activity compared to PAW and PAG, except for L. monocytogenes, where 40 min activation time and treatment time exceeding 20 min was required for significantly higher reduction to occur. PAN exhibited greater antibacterial activity at higher NaCl concentrations, which was attributed to increased ionic strength and reactive chlorine species. Additionally, we evaluated the microbial mechanisms of PAN by assessing cellular damage and alterations. The common observation across the three pathogens was that PAN resulted in increased cell membrane damage, reduced intermembrane enzyme activity, higher intracellular ROS levels, and changes in zeta potential values, while DNA damage was observed only in PAN-treated L. monocytogenes. Furthermore, when PAW and PAN were stored for up to four weeks, PAN showed higher efficacy compared to PAW. 10% PAN was also effective against foodborne pathogens on mackerel, achieving log reductions of 3.62 for E. coli O157:H7, 4.62 for S. Typhimurium, and 3.18 for L. monocytogenes after a 20 min treatment without adversely affecting quality. Our results demonstrated the antibacterial activity and action mechanism of PAN, presenting its potential application in the seafood industry.