RESUMEN
This study aims to evaluate the behaviour of Listeria monocytogenes under fluctuating temperature comparing the efficacy of deterministic and stochastic methods for its prediction. In the first part of the study, a strain of L. monocytogenes was maintained at two different fluctuating temperature regimes both from 2 to 8°C and regularly sampled for the quantitative determination. The first temperature regime lasted 204 hours with a fluctuation length of 12 hours whereas the second lasted 167 hours with a fluctuation length of 24 hours. A dynamic predictive model was implemented for the reproduction of the observed data. Model resolution has been carried out by using values of the recorded temperature as well as the value of the mean temperature, the kinetic mean temperature, the 75th and 95th percentile of the temperature. A stochastic resolution was also performed considering the mean temperature and Standard Deviation as stochastic variable. In the second part of the study, a temperature mean curve was constructed by monitoring temperature of 8 refrigerated conveyances, 10 display cabinet and 15 domestic refrigerators. This curve was used to obtain predictive scenarios for L. monocytogenes based on the above and also considering temperature regime suggested by the EURL Lm TECHNICAL GUIDANCE DOCUMENT on challenge tests and durability studies for assessing shelf-life of ready-to-eat foods related to Listeria monocytogenes (Version 4 of 1 July 2021). All predicted behaviours were compared to the observed ones through the Root Mean Squared Error. Firstly, dynamic predictive model as well as the stochastic one, provided the best level of reproducibility of the observed data. The kinetic mean temperature reproduced the observed data better than the mean temperature for the 12 hoursregime while for the 24 hours-regime was the opposite. The 75th and 95th percentile overestimated the observed growths. Secondary, predictions obtained with the mean temperature, kinetic temperature and stochastic approach well fitted the observed data. The 75th and 95th percentile of Temperature and the "Eurl LM" temperature regimes overestimated the observed prediction. Dynamic approach as well as the stochastic one allowed to obtain the lowest values of Root Mean Squared Error. The mean temperature and kinetic mean temperature appeared the most representative values in a deterministic "single-point" approach.
RESUMEN
Domestic environment, in particular, kitchen setting is a well-established source of microbial contamination. Kitchen sponges represent an important vehicle of microbial transmission and maintenance of spoilage bacteria and pathogenic strains responsible for food borne diseases. The aim of this study was to evaluate the microbial communities of 100 'in-use' kitchen sponges, improving the knowledge on their role in cross-contamination in domestic environment and transmission of ESBLproducing strains. Sponges were processed for: aerobic mesophilic bacteria (AMB), Enterobacteriaceae (EB), yeasts and molds (YM), coagulase-positive staphylococci (CPS), micrococci (MCC), anaerobic sulfite reducing bacteria (ASR), and for the detection of Listeria monocytogenes, Salmonella spp. and Yersinia enterocolitica. A total of 309 enterobacteria strains were identified and then processed for ESBL (Extended Spectrum Beta Lactamase) phenotypical expression. A high contamination level of kitchen sponges was observed (mean value AMB 8.25±1.1; EB 5.89±1.2; YM 5.57±1.1; MCC 4.82±0.1 log CFU/g). Identified enterobacteria strains revealed several opportunistic and pathogenic agents such as Enterobacter cloacae (28%), Citrobacter freundii (23.3%), Cronobacter sakazakii (14.6%) and other strains in lower percentage. Listeria monocytogenes was found in only one sponge (1%). A total of 69 (22.3%) enterobacteria resulted ESBL+, with the following prevalence: P. rettgeri (50%), L. adenocarboxilata (30%), K. pneumoniae (25%), K. oxytoca (25%), C. sakazakii (20%), E. cloacae (20.7%), C. freundii (20.1%). Results confirm the potential role of kitchen sponges as vehicle for food-borne pathogens such as, C. sakazakii for the first time, infectious agents and spoilage microorganisms. The observed high contamination level and the presence of several ESBLs opportunistic pathogens, stresses the necessity to improve a proper education of the consumers on the effective treatment to reduce their microbial loads.
RESUMEN
OBJECTIVE: To evaluate in vitro effects of Tagetes minuta L. essential oil (TEO) on L3 Anisakis larvae type 1. METHODS: In order to evaluate the potential use of Tagetes minuta essential oil against L3 Anisakis larvae three different media were tested: 1) a saline solution (SS); 2) an industrial marinating solution (MS); 3) sunflower seeds oil (SO). For each media and concentrations of TEO (0.1%, 0.5%, 1.0% and 5.0% v/v), 20 parasites were introduced into plastic Petri dishes (diameter 90 mm) and maintained at room temperature. As controls, larvae were maintained without TEO under identical experimental conditions in SS, MS and SO. A total of 900 larvae were tested. The normalized mean viability, LT100, LT50 and the percentage of inactivation at 24 h were calculated. RESULTS: In vitro tests revealed a complete inactivation of parasites in saline solution after 2 h with 5% and 1% of TEO. In marinating solution, a complete inactivation of parasites was observed after 4 h at all concentrations used. A slower activity for all TEO concentration was reported in SO. CONCLUSIONS: The results obtained, showing a strong activity against Anisakis larvae, confirm TEO as a larvicidal agent in the treatment of human anisakidosis and in the industrial marinating process.
