Different assembly of acid and salt tolerance response in two dairy Listeria monocytogenes wild strains.

A lack on the association between acid tolerance response (ATR) and osmotolerance response (OTR) among Listeria monocytogenes dairy isolates was found. In order to evaluate how wild L. monocytogenes isolates mount tolerance responses under a sub-lethal pH and a low sodium chloride concentration (pH 5.5 and 3.5 % [w/v] NaCl), a proteomic approach was used. The ATR and OTR of two L. monocytogenes cheese dairy isolates (strain T8, serotype 4b and A9, serotype 1/2b or 3b) were determined. The proteomes of the adapted and non-adapted cultures were evaluated by 2-DE. One strain displayed an ATR, but not an OTR and the other displayed an OTR, but not an ATR. The ATR positive strain showed the over-production of proteins related with protein synthesis, protein folding, attainment of reduction power, ribose production and cell wall. In contrast, in the OTR-positive-strain proteins related with glycolysis, general stress and detoxification were identified.

Proteomic analysis shows that individual Listeria monocytogenes strains use different strategies in response to gastric stress.

Ingestion of contaminated dairy products, in particular soft cheese, is one of the major routes of infection by the human pathogen Listeria monocytogenes. During cheese processing, this foodborne pathogen is exposed to sublethal acid and osmotic stress conditions, which may induce tolerance responses and influence subsequent survival in the gastric tract. The aim of the current study was to evaluate the impact on a L. monocytogenes cheese isolate (serotype 4b) and two cheese dairy isolates (T8, serotype 4b, isolated from vat; and A9, serotype 1/2b or 3b, isolated from shelf stand) of exposure to sublethal conditions of pH and salt (5.5 and 3.5% [w/v] NaCl) in a cheese-simulated medium and further challenge with gastric stress. The bacterial cells exposed to pH 7.0 and no added salt were considered non-adapted. Via two-dimensional gel electrophoresis (2-DE), the proteomes of cheese-simulated medium and gastric challenged Listeria cells were compared. All L. monocytogenes isolates were able to survive the high acidity of gastric fluid (pH 2.5), and no significant differences were observed between adapted and non-adapted cells. However, the analysis of the intracellular proteome profiles revealed a significant intra-strain variation in the protein arsenal used to respond to the adaptation in the cheese-based medium and to the gastric stress. In cheese-based medium, the three strains produced different stress proteins. All three strains showed a higher abundance of carbohydrate proteins, but there was no overlap between them. Exposure to the gastric fluid induced the production of a group of proteins in T8 adapted and non-adapted cells that had not been detected previously in the cheese-based proteome. No such response was shown by A9 and C882 strains. Taken together, this study evidences the proteome tools used by adapted and non-adapted cells to cope with the hostile microenvironment of the stomach.

Minimally processed fruits as vehicles for foodborne pathogens.

The consumption of minimally processed fruit (MPF) has increased over the last decade due to a novel trend in the food market along with the raising consumers demand for fresh, organic, convenient foods and the search for healthier lifestyles. Although represented by one of the most expanded sectors in recent years, the microbiological safety of MPF and its role as an emergent foodborne vehicle has caused great concern to the food industry and public health authorities. Such food products may expose consumers to a risk of foodborne infection as they are not subjected to prior microbial lethal methods to ensure the removal or destruction of pathogens before consumption. A considerable number of foodborne disease cases linked to MPF have been reported and pathogenic strains of Salmonella enterica, Escherichia coli, Listeria monocytogenes, as well as Norovirus accounted for the majority of cases. Microbial spoilage is also an issue of concern as it may result in huge economic losses among the various stakeholders involved in the manufacturing and commercialization of MPF. Contamination can take place at any step of production/manufacturing and identifying the nature and sources of microbial growth in the farm-to-fork chain is crucial to ensure appropriate handling practices for producers, retailers, and consumers. This review aims to summarize information about the microbiological hazards associated with the consumption of MPF and also highlight the importance of establishing effective control measures and developing coordinated strategies in order to enhance their safety.

Microbial quality of edible seeds commercially available in southern Portugal.

In the present work, the microbiological quality of sesame, flaxseed, chia, pumpkin sunflower seeds, a mix of seeds, as well as flaxseed flour, marketed in southern Portugal, were studied through the counting of aerobic microorganisms at 30 °C (AM), molds and yeast (M&Y), Escherichia coli (ß-glucuronidase positive) (ß-GP E. coli), Staphylococcus coagulase positive, and detection of Salmonella spp. The persistence of AM and M&Y populations were also counted in organic and non-organic flaxseed at 20 °C for 11 months. The seeds with the highest average of AM were flaxseed (1.3 x 106 CFU/g) followed by flaxseed flour (1.1 x 106 CFU/g) while the lowest level was found in chia (2.9 x 104 CFU/g). This seed also presented the lowest average values of filamentous fungi (9.8 x 102 CFU/g), whereas sunflower seeds had the highest levels (1.7 x 105 CFU/g). Flaxseed flour had the highest yeast counts (1.5 x 104 CFU/g). Although some samples had high levels of AM and fungi, ß-GP E. coli and Salmonella were not detected, therefore, they complied with the microbiological criteria of the European Union. The organic flaxseed contained higher numbers of AM and M&Y than the non-organic ones (p < 0.05). In addition, the storage of flaxseed at 20 °C resulted in changes of AM and M&Y, showing that these populations were able to remain viable after eleven months (AM Log 5.4-Log 5.6; M&Y Log 2.8-Log 4.1). The results obtained in the present study, namely those high levels of AM and fungi (>106 and 104 CFU/g respectively), alert to the need of improving processing practices, storage/distribution conditions of edible seeds and derivatives, as well as the requirement of implementing adequate decontamination techniques.