Pre-Exposure of Foodborne Staphylococcus aureus Isolates to Organic Acids Induces Cross-Adaptation to Mild Heat

Staphylococcus aureus is a typical enterotoxin-producing bacterium that causes food poisoning. In the food industry, pasteurization is the most widely used technique for food decontamination. However, pre-exposure to an acidic environment might make bacteria more resistant to heat treatment, which could compromise the bactericidal effect of heat treatment and endanger food safety. In this work, the organic acid-induced cross-adaptation of S. aureus isolates to heat and the associated mechanisms were investigated. Cross-adaptation area analysis indicated that pre-exposure to organic acids induced cross-adaptation of S. aureus to heat in a strain-dependent manner. Compared with other strains, S. aureus strain J15 showed extremely high heat resistance after being stressed by acetic acid, citric acid, and lactic acid. S. aureus strains J19, J9, and J17 were found to be unable to develop cross-adaptation to heat with pre-exposure to acetic acid, citric acid, and lactic acid, respectively. Analysis of the phenotypic characteristics of the cell membrane demonstrated that the acid-heat-cross-adapted strain J15 retained cell membrane integrity and functions through enhanced Na(+)K(+)-ATPase and F(o)F(1)-ATPase activities. Cell membrane fatty acid analysis revealed that the ratio of anteiso to iso branched-chain fatty acids in the acid-heat-cross-adapted strain J15 decreased and the content of straight-chain fatty acids exhibited a 2.9 to 4.4% increase, contributing to the reduction in membrane fluidity. At the molecular level, fabH was overexpressed with preconditioning by organic acid, and its expression was further enhanced with subsequent heat exposure. Organic acids activated the GroESL system, which participated in the heat shock response of S. aureus to the subsequent heat stress. IMPORTANCE Cross-adaptation is one of the most important phenotypes in foodborne pathogens and poses a potential risk to food safety and human health. In this work, we found that pretreatment with acetic acid, citric acid, and lactic acid could induce subsequent heat tolerance development in S. aureus. Various S. aureus strains exhibited different acid-heat cross-adaptation areas. The acid-induced cross-adaptation to heat might be attributable to membrane integrity maintenance, stabilization of the charge equilibrium to achieve a normal internal pH, and membrane fluidity reduction achieved by decreasing the ratios of anteiso to iso fatty acids. The fabH gene, which is involved in fatty acid biosynthesis, and groES/groEL, which are related to heat shock response, contributed to the development of the acid-heat cross-adaptation phenomenon in S. aureus. The investigations of the stress cross-adaptation phenomenon in foodborne pathogens could help optimize food processing to better control S. aureus.