Formation of Sublethally Injured Yersinia enterocolitica, Escherichia coli O157:H7, and Salmonella enterica Serovar Enteritidis Cells after Neutral Electrolyzed Oxidizing Water Treatments

Neutral electrolyzed oxidizing (NEO) water is a nonthermal and less corrosive antimicrobial treatment that has been demonstrated to have efficacy in reducing microbial contamination in food, including meat, fresh fruit, and vegetables. However, NEO water treatments can cause sublethal injury to pathogenic cells, resulting in cells that retain their viability. Consequently, these sublethally injured pathogenic cells become a serious food safety concern. This study evaluated the formation of sublethally injured Yersinia enterocolitica, Escherichia coli O157:H7, and Salmonella enterica serovar Enteritidis cells by NEO water treatments and the potential cross protection against heat, pH, NaCl, or bile salt stresses that it may generate. No cross protection was observed. By combining NEO water treatments with sublethal levels of additional stresses, significant synergistic antimicrobial outcomes were achieved. These results indicate that mild processing treatments, when combined, can effectively reduce pathogen populations while minimizing the negative impacts on food quality. ABSTRACT The impact of neutral electrolyzed oxidizing (NEO) water treatments on the formation of sublethally injured Yersinia enterocolitica, Escherichia coli O157:H7, and Salmonella enterica serovar Enteritidis cells was evaluated. When pathogens were treated with 6% NEO water, approximately 38% of the treated Yersinia population and 25% of the treated Salmonella population became sublethally injured. The highest sublethally injured population was found when Salmonella cultures were treated with 3% NEO water. Regardless of the NEO water concentration used, no sublethally injured E. coli O157:H7 cells were found. To evaluate the sensitivity of NEO water-treated cells, four additional stresses (heat treatment, pH, NaCl, and bile salt) were tested. NEO water treatments did not generate any cross protection of treated cells against the other stresses. The diluted NEO water treatments in combination with heat treatment at 51°C for 10 min led to the best synergistic antimicrobial effects with a combined reduction of 7 logs. The gene expression results showed that NEO water treatments led to the upregulation of ompR, ail, and ycfR. These genes are known for their involvement in cells' environmental stress responses. In summary, this study investigated the sublethal injury in pathogenic cells caused by NEO water treatments. Although sublethal injury was discovered, when combined with other mild stresses, the synergistic antimicrobial effects were able to further reduce the numbers of viable pathogenic cells. These results demonstrate the great application potential of NEO water as a nonthermal and less corrosive antimicrobial treatment. IMPORTANCE Neutral electrolyzed oxidizing (NEO) water is a nonthermal and less corrosive antimicrobial treatment that has been demonstrated to have efficacy in reducing microbial contamination in food, including meat, fresh fruit, and vegetables. However, NEO water treatments can cause sublethal injury to pathogenic cells, resulting in cells that retain their viability. Consequently, these sublethally injured pathogenic cells become a serious food safety concern. This study evaluated the formation of sublethally injured Yersinia enterocolitica, Escherichia coli O157:H7, and Salmonella enterica serovar Enteritidis cells by NEO water treatments and the potential cross protection against heat, pH, NaCl, or bile salt stresses that it may generate. No cross protection was observed. By combining NEO water treatments with sublethal levels of additional stresses, significant synergistic antimicrobial outcomes were achieved. These results indicate that mild processing treatments, when combined, can effectively reduce pathogen populations while minimizing the negative impacts on food quality.