Effect of Hydrogen Peroxide and Sodium Chloride on Enumeration of Thermally Stressed Cells of Staphylococcus aureus

Abstract

Evidence is presented suggesting that the decreased enumeration of heat-stressed Staphylococcus aureus cells on selective media is the result of accumulation of metabolic H2O2. It accumulates due to the decreased activity of catalase caused by the synergistic effects of heat and NaCl. Heated cells enumerated anaerobically on tryptic soy agar (TSA) containing 6.5% NaCl (TSAS 6.5) exhibited a 200-fold increase compared to cells enumerated aerobically on the same medium. The anaerobic counts on TSAS 6.5 were similar to the aerobic counts on TSA. Increases in both death and injury occurred when S. aureus was propagated in tryptic soy broth (TSB) plus 10% NaCl (TSBS) instead of TSB before thermal injury. Addition of catalase to TSA and TSA containing 7.5% NaCl (TSAS) increased the count to approximately the same levels on TSA and TSAS as that found following thermal injury after propagation in TSB. Catalase activity was 12-fold higher in stationary phase cells propagated in TSB than in TSBS. Indirect evidence indicates that toxic levels of H2O2 accumulated rapidly, causing one to two log decreases in enumeration after 30 to 60 min incubation on TSAS.