Impact of environmental factors on the culturability and viability of Listeria monocytogenes under conditions encountered in food processing plants.

The ability of Listeria monocytogenes to adhere to and persist on surfaces for months or even years may be responsible for its transmission from contaminated surfaces to food products. Hence the necessity to find effective means to prevent the establishment of L. monocytogenes in food processing environments. The aim of this study was to assess, through a fractional experimental design, the environmental factors that could affect the survival of L. monocytogenes cells on surfaces to thereby prevent the persistence of this pathogen in conditions mimicking those encountered in food processing plants: culture with smoked salmon juice or meat exudate, use of two materials with different hygiene status, biofilm of L. monocytogenes in pure-culture or dual-culture with a Pseudomonas fluorescens strain, application of a drying step after cleaning and disinfection (C&D) and comparison of two strains of L. monocytogenes. Bacterial survival was assessed by culture, qPCR to quantify total cells, and propidium monoazide coupled with qPCR to quantify viable cells and highlight viable but non-culturable (VBNC) cells. Our results showed that failure to apply C&D causes cell persistence on surfaces. Moreover, the sanitation procedure leads only to a loss of culturability and appearance of VBNC populations. However, an additional daily drying step after C&D optimises the effectiveness of these procedures to reduce culturable populations. Our results reinforce the importance to use molecular tools to monitor viable pathogens in food processing plants to avoid underestimating the amounts of cells using only methods based on cell culture.

Development of synthetic media mimicking food soils to study the behaviour of Listeria monocytogenes on stainless steel surfaces.

Listeria monocytogenes is one of the main targets of hygiene procedures in the ready-to-eat food industry due to its ability to persist for months or even years in processing plants, where it can contaminate food during processing. The factors associated with persistence are often those that foster growth, which itself depends on food contamination of surfaces. It is therefore essential to experiment by using food soils or media modelling these soils to understand the behaviour of L. monocytogenes on surfaces of food processing plants. Thus, we set up an experimental plan including three physiological parameters characteristic of the behaviour of cells on surfaces, namely spatial distribution, adhesion forces and the physiological state of sessile L. monocytogenes. These were recorded in two food soils: smoked salmon juice and meat exudate. According to our results, the behaviour of L. monocytogenes on stainless steel surfaces is highly dependent on the food soil used. The presence of viable but non-culturable (VBNC) cells was demonstrated using meat exudate, while all viable cells were recovered using smoked salmon juice. Moreover, on the basis of our criteria and after validation with three strains of L. monocytogenes, we showed that smoked salmon juice can be substituted by a modified culture medium, demonstrating that drawbacks associated with the use of food soils can be overcome.