Consumption of minimally-processed, or fresh-cut, fruit and vegetables has rapidly increased in recent years, but there have also been several reported outbreaks associated with the consumption of these products. Sodium hypochlorite is currently the most widespread disinfectant used by fresh-cut industries. Neutral electrolyzed water (NEW) is a novel disinfection system that could represent an alternative to sodium hypochlorite. The aim of the study was to determine whether NEW could replace sodium hypochlorite in the fresh-cut produce industry. The effects of NEW, applied in different concentrations, at different treatment temperatures and for different times, in the reduction of the foodborne pathogens Salmonella, Listeria monocytogenes and Escherichia coli O157:H7 and against the spoilage bacterium Erwinia carotovora were tested in lettuce. Lettuce was artificially inoculated by dipping it in a suspension of the studied pathogens at 10(8), 10(7) or 10(5) cfu ml(-1), depending on the assay. The NEW treatment was always compared with washing with deionized water and with a standard hypochlorite treatment. The effect of inoculum size was also studied. Finally, the effect of NEW on the indigenous microbiota of different packaged fresh-cut products was also determined. The bactericidal activity of diluted NEW (containing approximately 50 ppm of free chlorine, pH 8.60) against E. coli O157:H7, Salmonella, L. innocua and E. carotovora on lettuce was similar to that of chlorinated water (120 ppm of free chlorine) with reductions of 1-2 log units. There were generally no significant differences when treating lettuce with NEW for 1 and 3 min. Neither inoculation dose (10(7) or 10(5) cfu ml(-1)) influenced the bacterial reduction achieved. Treating fresh-cut lettuce, carrot, endive, corn salad and 'Four seasons' salad with NEW 1:5 (containing about 50 ppm of free chlorine) was equally effective as applying chlorinated water at 120 ppm. Microbial reduction depended on the vegetable tested: NEW and sodium hypochlorite treatments were more effective on carrot and endive than on iceberg lettuce, 'Four seasons' salad and corn salad. The reductions of indigenous microbiota were smaller than those obtained with the artificially inoculated bacteria tested (0.5-1.2 log reduction). NEW seems to be a promising disinfection method as it would allow to reduce the amount of free chlorine used for the disinfection of fresh-cut produce by the food industry, as the same microbial reduction as sodium hypochlorite is obtained. This would constitute a safer, 'in situ', and easier to handle way of ensuring food safety.
Fate of Listeria monocytogenes on fully ripened Greek Graviera cheese stored at 4, 12, or 25 degrees C in air or vacuum packages: in situ PCR detection of a cocktail of bacteriocins potentially contributing to pathogen inhibition.
The behavior of Listeria monocytogenes on fully ripened Greek Graviera cheese was evaluated. Three batches (A, B, and C) were tested. Batches A and C were prepared with a commercial starter culture, while in batch B the starter culture was combined with an enterocin-producing Enterococcus faecium Graviera isolate. Cheese pieces were surface inoculated with a five-strain cocktail of L. monocytogenes at ca. 3 log CFU/cm2, packed under air or vacuum conditions, stored at 4, 12, or 25 degrees C, and analyzed after 0, 3, 7, 15, 30, 60, and 90 days. L. monocytogenes did not grow on the cheese surface, regardless of storage conditions. However, long-term survival of the pathogen was noted in all treatments, being the highest (P
Five grams of seafood products were inoculated with one to 500 viable or 10(9) heat-killed cells of Listeria monocytogenes. The presence of the pathogen was detected by the polymerase chain reaction (PCR) with primers specific for fragments of the listeriolysin O (hly) gene (two sets) and for the invasion-associated protein (iap) gene (one set). For DNA preparation, boiling, either alone or in combination with lysozyme and proteinase K treatment, was not always sufficient to lyse L. monocytogenes, while treatment with Triton X-100 produced consistently good DNA suitable for amplification. To avoid false-negative and false-positive results, 48 h incubations were necessary and a subculturing step after an initial 24 h incubation greatly improved the results. The primers that amplified regions of the listeriolysin O gene gave clearer and stronger products than primers for the invasion-associated protein gene. Using this method we were able to detect one to five L. monocytogenes cells in 5 g of product in a total of 55 h.