Currently, there is a growing preference for convenience food products, such as ready-toeat (RTE) foods, associated with long refrigerated shelf-lives, not requiring a heat treatment prior to consumption. Because Listeria monocytogenes is able to grow at refrigeration temperatures, inconsistent temperatures during production, distribution, and at consumer's household may allow for the pathogen to thrive, reaching unsafe limits. L. monocytogenes is the causative agent of listeriosis, a rare but severe human illness, with high fatality rates, transmitted almost exclusively by food consumption. With the aim of assessing the quantitative microbial risk of L. monocytogenes in RTE chicken salads, a challenge test was performed. Salads were inoculated with a three-strain mixture of cold-adapted L. monocytogenes and stored at 4, 12, and 16°C for eight days. Results revealed that the salad was able to support L. monocytogenes' growth, even at refrigeration temperatures. The Baranyi primary model was fitted to microbiological data to estimate the pathogen's growth kinetic parameters. Temperature effect on the maximum specific growth rate (μ max) was modeled using a square-root-type model. Storage temperature significantly influenced μ max of L. monocytogenes (p < 0.05). These predicted growth models for L. monocytogenes were subsequently used to develop a quantitative microbial risk assessment, estimating a median number of 0.00008726 listeriosis cases per year linked to the consumption of these RTE salads. Sensitivity analysis considering different time-temperature scenarios indicated a very low median risk per portion (<−7 log), even if the assessed RTE chicken salad was kept in abuse storage conditions.
Listeriosis is almost entirely transmitted through foods contaminated with Listeria monocytogenes. Ready-to-eat foods present a particular challenge due to their long refrigerated shelf-life, not requiring any heat treatment before consumption. In this work, a shelf-life assessment of an industrially produced ready-to-eat salad was performed using conventional culture-based and molecular methods. L. monocytogenes isolates were confirmed and serogrouped using multiplex PCR, and genetic subtyping was performed by pulsed-field gel electrophoresis (PFGE). PMAxx-qPCR was used as an alternative method for L. monocytogenes quantification in foods. Salad samples were kept at 4 °C, 12 °C, and 16 °C for eight days and analysed. At 4 °C, acceptable results were obtained considering hygiene indicators, i.e., Enterobacteriaceae (ranging from 3.55 ± 0.15 log cfu/g to 5.39 ± 0.21 log cfu/g) and aerobic mesophilic colony counts (5.91 ± 0.90 log cfu/g to 9.41 ± 0.58 log cfu/g) throughout the study, but the same did not happen at 12 °C and 16 °C. L. monocytogenes culture-based quantification exhibited low numbers (<1 log cfu/g) for all temperatures. From 30 presumptive isolates, 10 (33.3%) were confirmed as L. monocytogenes with the majority belonging to serogroup IVb. PFGE subtyping showed that 7 of the 10 L. monocytogenes isolates had 100% of pulsotype similarity, suggesting a possible common contamination source. PMAxx-qPCR revealed a statistically higher L. monocytogenes quantification (>3 log cfu/g) when compared to the conventional culture-based method, suggesting viable but non-culturable forms. Taken together, results underline the need to combine conventional methods with more sensitive, specific, and rapid ones for L. monocytogenes assessment in ready-to-eat foods shelf-life studies to reduce the potential risk for consumers.
Listeria Monocytogenes is an important foodborne pathogen with the capacity to grow at low temperatures and the ability to form biofilms. These features are particularly significant to food business operators producing readyto-eat foods with a long refrigerated shelf-life not undergoing any listericidal treatment before consumption. Objectives: This work aims to assess the temperature effect on L. monocytogenes growth in planktonic suspension and in mono-species biofilms. Methods and results: Isothermal planktonic growth at 12o C and 37o C was assayed using viable cell counts and optical density measurements that revealed a strong positive correlation, confirming the reliability of combining both methods to estimate L. monocytogenes concentration. Experimental data were then fitted to Baranyi and Roberts primary predictive model and the estimated growth parameters confirmed that μmax at 37o C (0.375 ± 0.072 log Cfu/ ml/h) was higher than at 120 C (0.054 ± 0.001 log Cfu/ml/h), with identical L. monocytogenes final concentrations which emphasizes its ability to grow at refrigerated temperatures. Experimental results from the isothermal growth assay and ComBase Predictor growth model were similar, with slightly higher estimated μmax (37o C: 0.480 log Cfu/ml/h; 12o C: 0.068 log Cfu/ml/h) in the predictor growth model. The studied strains demonstrated biofilm-forming ability at 12o C, 20o C and 300 C after 5 days of growth. No significant differences in biofilm formation at different temperatures were detected considering viable cell counts values, but when using crystal violet staining optical density results significant differences were found, with the highest formation occurring at 30ºC. A positive strong correlation was found between viable cell counts and crystal violet staining optical density results. In fact, both methods complement each other, because while viable cell counts measures viable cells, crystal violet staining optical density considers total biomass (viable and non-viable cells and extracellular matrix components). Nevertheless, in this work all L. monocytogenes strains revealed to be weak biofilm producers. Conclusion: Overall, this studys results contribute with important initial information on L. monocytogenes growth and biofilm formation to further assist predictive growth modeling in food matrices and environments, also enabling subsequent quantitative microbial risk assessment, to improve pathogen’s control.
The biomechanical properties of the human skin are intrinsically correlated with changes associated with pathological conditions, aging, and hydration. Quantitative measurements can improve diagnostic tools, treatments, and cosmetic product evaluation. Using optical coherence elastography (OCE), an emerging imaging modality combining optical coherence tomography (OCT) with a localized excitation source to induce mechanical disturbances, a quantitative evaluation of tissue biomechanics can be achieved. OCE complements the structural information with elasticity data to attain a complete overview of skin status. In this study, we employed a home-built OCE system, combining a swept-source OCT system with a piezoelectric actuator for tissue displacement, to evaluate changes to the skin biomechanical properties due to the application of an anti-aging cream. Skin elasticity was monitored for a total of five weeks. Anti-aging cream was applied daily for four weeks. OCE measurements continued for one additional week to assess the effect of cream application interruption. Three female volunteers were included in this proof-of-principle investigation. Their counter-arm was used as control. Although no statistical significance was reached, a decrease in skin Young’s modulus was observed with the cream application, indicating an increase in skin elasticity.
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