Multi-method approach indicates no presence of sub-lethally injured Listeria monocytogenes cells after mild heat treatment

Application of mild inactivation treatments follows an increasing trend in the food industry and is often combined with sub-optimal intrinsic product conditions to ensure appropriate level of microbial safety. Listeria monocytogenes was subjected to mild heat treatment (20 min at 60 °C) and subsequently exposed to various mild preservation conditions based on increased NaCl concentration and decreased pH. Recovery and resuscitation of L. monocytogenes cells were studied using various methods. Using 12-fold Most Probable Number (MPN) method no difference in the amount of recovered cells under adverse conditions was noted between heat-treated and non-treated L. monocytogenes cells. Time-to-detection method using on-line OD measurements showed that heat-treated L. monocytogenes cells reached detection limit faster in acidified media and NaCl supplemented media in comparison with non-heated control cells.Flow cytometry (FCM) analysis using 5–6-carboxyfluorescein diacetate (cFDA) and propidium iodide (PI) staining showed presence of low numbers of viable cells. Overall, there was no indication of sub-lethal injury in L. monocytogenes cells after mild heat treatment.