RESUMEN

Understanding the role of food-related factors on the efficacy of protective cultures is essential to attain optimal results for developing biopreservation-based strategies. The aim of this work was to assess and model growth of Latilactobacillus sakei CTC494 and Listeria monocytogenes CTC1034, and their interaction, in two different ready-to-eat fish products (i.e., surimi-based product and tuna pâté) at 2 and 12 °C. The existing expanded Jameson-effect and a new expanded Jameson-effect model proposed in this study were evaluated to quantitatively describe the effect of microbial interaction. The inhibiting effect of the selected lactic acid bacteria strain on the pathogen growth was product dependent. In surimi product, a reduction of lag time of both strains was observed when growing in coculture at 2 °C, followed by the inhibition of the pathogen when the bioprotective L. sakei CTC494 reached the maximum population density, suggesting a mutualism-antagonism continuum phenomenon between populations. In tuna pâté, L. sakei CTC494 exerted a strong inhibition of L. monocytogenes at 2 °C (<0.5 log increase) and limited the growth at 12 °C (<2 log increase). The goodness-of-fit indexes indicated that the new expanded Jameson-effect model performed better and appropriately described the different competition patterns observed in the tested fish products. The proposed expanded competition model allowed for description of not only antagonistic but also mutualism-based interactions based on their influence on lag time.

Asunto(s)

RESUMEN

The occurrence of various foodborne disease outbreaks linked to the consumption of cucumbers worldwide in the last years raised concerns regarding the survival ability of foodborne pathogens on this food matrix. This work aimed at evaluating and quantifying the survival of Escherichia coli O157:H7 and Salmonella spp. on cucumber surfaces. Cucumbers were inoculated with a 5-strain cocktail of each microorganism and kept at 25 °C. The survival ability of two green fluorescent protein (GFP) labelled Salmonella strains inoculated individually on cucumbers was also evaluated. The inoculated areas were swabbed at different time intervals (maximum of 72 h) and cells were enumerated by plate count method (log CFU/cm2). The population of both pathogens decreased significantly on cucumber surfaces over time. E. coli O157:H7 could only be recovered up to 8 h while Salmonella spp. could be detected up to 24 h. The GFP-labelled Salmonella strains showed similar behaviour on cucumbers compared to the evaluated Salmonella cocktail. Survival kinetic parameters were estimated by fitting the Weibull model to the survival data. The data obtained in this study indicate that despite of the rapid decrease on concentrations of both pathogens evaluated on cucumbers surfaces, strategies to avoid their contamination during the supply chain as well as proper cleaning and disinfection protocols must be put forward to mitigate both E. coli O57:H7 and Salmonella on cucumbers and therefore, to decrease the exposure of consumers to microbial hazards and to avoid cross-contamination events during distribution, retail and in domestic environments.

Asunto(s)

RESUMEN

In this study, the bioprotective potential of Lactobacillus sakei CTC494 against Listeria monocytogenes CTC1034 was evaluated on vacuum packaged hot-smoked sea bream at 5 °C and dynamic temperatures ranging from 3 to 12 °C. The capacity of three microbial competition interaction models to describe the inhibitory effect of L. sakei CTC494 on L. monocytogenes was assessed based on the Jameson effect and Lotka-Volterra approaches. A sensory analysis was performed to evaluate the spoiling capacity of L. sakei CTC494 on the smoked fish product at 5 °C. Based on the sensory results, the bioprotection strategy against the pathogen was established by inoculating the product at a 1:2 ratio (pathogen:bioprotector, log CFU/g). The kinetic growth parameters of both microorganisms were estimated in mono-culture at constant storage (5 °C). In addition, the inhibition function parameters of the tested interaction models were estimated in co-culture at constant and dynamic temperature storage using as input the mono-culture kinetic parameters. The growth potential (δ log) of L. monocytogenes, in mono-culture, was 3.5 log on smoked sea bream during the experimental period (20 days). In co-culture, L. sakei CTC494 significantly reduced the capability of L. monocytogenes to grow, although its effectiveness was temperature dependent. The LAB strain limited the growth of the pathogen under storage at 5 °C (<1 log increase) and at dynamic profile 2 (<2 log increase). Besides, under storage at dynamic profile 1, the growth of L. monocytogenes was inhibited (<0.5 log increase). These results confirmed the efficacy of L. sakei CTC494 for controlling the pathogen growth on the studied fish product. The Lotka-Volterra competition model showed slightly better fit to the observed L. monocytogenes growth response than the Jameson-based models according to the statistical performance. The proposed modelling approach could support the assessment and establishment of bioprotective culture-based strategies aimed at reducing the risk of listeriosis linked to the consumption of RTE hot-smoked sea bream.

Asunto(s)

RESUMEN

In this study, the inhibitory capacity of Lactobacillus sakei strain L115 against Listeria monocytogenes has been assayed at 4, 8, 11, 15 and 20 °C in broth culture. Besides, the use of predictive microbiology models for describing growth of both microorganisms in monoculture and coculture has been proposed. A preliminary inhibitory test confirmed the ability of Lb. sakei strain L115 to prevent the growth of a five-strain cocktail of L. monocytogenes. Next, the growth of microorganisms in isolation, i.e. in monoculture, was monitored and kinetic parameters maximum specific growth rate (µsp;max) and maximum population density (Nmax) were estimated by fitting the Baranyi model to recorded data. Inhibition coefficients (α) were calculated for the two kinetic parameters tested (µsp:max and Nmax) to quantify the percentage of reduction of growth when the microorganisms were in coculture in comparison with monoculture. The kinetic parameters were input into three interaction models, developed based on modifications of the Baranyi growth model, namely Jameson effect, new modified version of the Jameson effect and Lotka-Volterra models. Two approaches were utilized for simulation, one using the monoculture µsp;max, under the hypothesis that the growth potential is similar under monoculture and coculture conditions provided the environmental conditions are not modified, and the other one, based on adjusting the monoculture kinetic parameter by applying the corresponding α to reproduce the observed µsp;max under coculture conditions, assuming, in this approach, that the existence of a heterogeneous population can change the growth potential of each microbial population. It was observed that in coculture, µsp;max of L. monocytogenes decreased (e.g., α = 31% at 4 °C) and the Nmax was much lower than that of monoculture (e.g., α = 36% at 4 °C). The best simulation performance was achieved applying α to adjust the estimated monoculture growth rate, with the modified Jameson and Lotka-Volterra models showing better fit to the observed microbial interaction data as demonstrated by the fact that 100% data points fell within the acceptable simulation zone (±0.5 log CFU/mL from the simulated data). More research is needed to clarify the mechanisms of interaction between the microorganisms as well as the role of temperature.

Asunto(s)

RESUMEN

This study was aimed at characterizing microbiologically Gilthead sea bream (Sparus aurata) and Sea bass (Dicentrarchus labrax) produced in two estuarine ecosystems in Andalusia (Spain): the estuary of the river Guadalquivir (La Puebla del Río, Sevilla) (A), and the estuary of the river Guadiana (Ayamonte, Huelva) (B). The collected fish individuals and water were analysed for hygiene indicator microorganisms and pathogens. The statistical analysis of results revealed that microbial counts for the different microbiological parameters were not statistically different for fish type. On the contrary, considering anatomic part, viscera showed significantly higher concentrations for Enterobacteriaceae, total coliforms and for Staphylococcus spp. coagulase +. Furthermore, location A showed in water and fish higher levels for lactic acid bacteria, aerobic mesophilic bacteria, Enterobacteriaceae, total coliforms and Staphylococcus spp. coagulase +. Neither Listeria monocytogenes, nor Salmonella spp. were detected, though Vibrio parahaemolyticus was identified, molecularly, in estuarine water in location B. The predictive analysis demonstrated that the initial microbiological quality could have an impact on product shelf-life, being longer for location B, with better microbiological quality. Results stress the relevance of preventing the microbiological contamination of water in estuary production systems in order to assure the quality and safety of Gilthead sea bream and Sea bass.

Asunto(s)

RESUMEN

The objective of this work was to quantitatively evaluate the effect of Lactobacillus sakei CTC494 (sakacin-producing bioprotective strain) against Listeria monocytogenes in fish juice and to apply and validate three microbial interaction models (Jameson, modified Jameson and Lotka Volterra models) through challenge tests with gilthead sea bream (Sparus aurata) fillets under modified atmosphere packaging stored at isothermal and non-isothermal conditions. L. sakei CTC494 inhibited L. monocytogenes growth when simultaneously present in the matrix (fish juice and fish fillets) at different inoculation ratios pathogen:bioprotector (i.e. 1:1, 1:2 and 1:3). The higher the inoculation ratio, the stronger the inhibition of L. monocytogenes growth, with the ratio 1:3 yielding no growth of the pathogen. The maximum population density (Nmax) was the most affected parameter for L. monocytogenes at all inoculation ratios. According to the microbiological and sensory analysis outcomes, an initial inoculation level of 4 log cfu/g for L. sakei CTC494 would be a suitable bioprotective strategy without compromising the sensory quality of the fish product. The performance of the tested interaction models was evaluated using the Acceptable Simulation Zone approach. The Lotka Volterra model showed slightly better fit than the Jameson-based models with 75-92% out of the observed counts falling into the Acceptable Simulation Zone, indicating a satisfactory model performance. The evaluated interaction models could be used as predictive modelling tool to simulate the simultaneous behaviour of bacteriocin-producing Lactobacillus strains and L. monocytogenes; thus, supporting the design and optimization of bioprotective culture-based strategies against L. monocytogenes in minimally processed fish products.

Asunto(s)

RESUMEN

A tool able to quantitatively assess the fate of potential pathogenic microorganisms in foods along the food chain and their impact on public health is highly valuable for food safety decision-makers. The aim of this work was to present an overview of the Predictive Microbiology software MicroHibro, which is able to assess the evolution of potential pathogens and spoilage microorganisms along the food chain, providing estimates for the exposure level and risk associated with a food product. The application is built on an extensive Predictive Microbiology Model Data Base (PMDB) including kinetic processes like growth, inactivation, transfer as well as dose-response models. PMDB can be populated with new models by using an on-line tool in combination with a standardized method for describing Predictive Microbiology models. This enables MicroHibro to be easily updated, increasing its applicability and use. Estimation of microbial risk associated with a food product can be achieved, in MicroHibro, by describing steps in any food chain using four different microbial processes (growth, inactivation, transfer and partitioning). As a result, an estimate of the concentration and prevalence of microorganisms in the food of interest as well as attendant risk are provided. Also, MicroHibro allows comparing different predictive models and validate them by introducing user's data. In this paper, examples are provided to illustrate how predictive models can be incorporated in MicroHibro, and then, used to develop a Quantitative Microbial Risk Assessment model. The use of expert computational systems is a powerful tool for supporting food safety and quality activities by Health Authorities and the food industry. They represent a breakthrough in the assessment and management of food safety based on scientific evidence.

Asunto(s)

RESUMEN

Over the last couple of decades, several studies have evaluated growth dynamics of L. monocytogenes in lightly processed and ready-to-eat (RTE) fishery products mostly consumed in Nordic European countries. Other fish species from aquaculture production are of special interest since their relevant consumption patterns and added value in Mediterranean countries, such as sea bream and sea bass. In the present study, the growth of L. monocytogenes was evaluated in fish-based juice (FBJ) by means of optical density (OD) measurements in a temperature range 2-20 °C under different atmosphere conditions (i.e. reduced oxygen and aerobic). The Baranyi and Roberts model was used to estimate the maximum growth rate (µmax) from the observed growth curves. The effect of storage temperature on µmax was modelled using the Ratkowsky square root model. The developed models were validated using experimental growth data for L. monocytogenes in sea bream and sea bass fillets stored under static and dynamic temperature conditions. Overall, models developed in FBJ provided fail-safe predictions for L. monocytogenes growth. For the model generated under reduced oxygen conditions, bias and accuracy factor for growth rate predictions were 1.15 and 1.25, respectively, showing good performance to adequately predict L. monocytogenes growth in Mediterranean fish products. The present study provides validated predictive models for L. monocytogenes growth in Mediterranean fish species to be used in microbial risk assessment and shelf-life studies.

Asunto(s)

RESUMEN

The purpose of this study was to evaluate and model the growth of Escherichia coli O157:H7 in fresh-cut lettuce submitted to a neutral electrolyzed water (NEW) treatment, packaged in passive modified atmosphere and subsequently stored at different temperatures (4, 8, 13, 16°C) for a maximum of 27 days. Results indicated that E. coli O157:H7 was able to grow at 8, 13, and 16°C, and declined at 4°C. However at 8°C, the lag time lasted 19 days, above the typical shelf-life time for this type of products. A secondary model predicting growth rate as a function of temperature was developed based on a square-root function. A comparison with literature data indicated that the growth predicted by the model for E. coli O157:H7 was again lower than those observed with other disinfection treatments or packaging conditions (chlorinated water, untreated product, NEW, etc.). The specific models here developed might be applied to predict growth in products treated with NEW and to improve existing quantitative risk assessments.

Asunto(s)

RESUMEN

Fresh-cut iceberg lettuce inoculated with Escherichia coli O157:H7 was submitted to chlorine washing (150 mg/mL) and modified atmosphere packaging on laboratory scale. Populations of E. coli O157:H7 were assessed in fresh-cut lettuce stored at 4, 8, 13 and 16 °C using 6-8 replicates in each analysis point in order to capture experimental variability. The pathogen was able to grow at temperatures ≥8 °C, although at low temperatures, growth data presented a high variability between replicates. Indeed, at 8 °C after 15 days, some replicates did not show growth while other replicates did present an increase. A growth primary model was fitted to the raw growth data to estimate lag time and maximum growth rate. The prediction and confidence bands for the fitted growth models were estimated based on Monte-Carlo method. The estimated maximum growth rates (log cfu/day) corresponded to 0.14 (95% CI: 0.06-0.31), 0.55 (95% CI: 0.17-1.20) and 1.43 (95% CI: 0.82-2.15) for 8, 13 and 16 °C, respectively. A square-root secondary model was satisfactorily derived from the estimated growth rates (R(2) > 0.80; Bf = 0.97; Af = 1.46). Predictive models and data obtained in this study are intended to improve quantitative risk assessment studies for E. coli O157:H7 in leafy green products.

Asunto(s)

RESUMEN

BACKGROUND: This study was conducted in eight Spanish school canteens during the period 2008-2009. Food handlers' practices, kitchen equipment, hygiene/sanitation conditions and handling practices were evaluated using checklists. In parallel, the microbiological quality and safety of ready-to-eat (RTE) vegetable salads were assessed. In addition, food contact surfaces and environmental air quality of different areas were analysed. The study determined the relationship between the microbiological quality of RTE foods and food handling practices, together with the degree of contamination of working surfaces and environmental contamination of processing and distribution areas. RESULTS: Some deficiencies were found regarding the use and change of gloves, hand-washing and cleanliness of working surfaces. The microbial levels detected in the foods examined indicated the absence of pathogens in the samples analysed. Surface counts were higher on cutting boards and faucets, showing insufficient cleanliness procedures. CONCLUSION: This study constitutes a descriptive analysis of the hygiene/sanitation conditions implemented in food service systems in eight Spanish school canteens. The results should help risk managers to better define control measures to be adopted in order to prevent foodborne infections.

Asunto(s)

RESUMEN

Foodborne diseases constitute a major concern in societies, and their causes are aimed to be identified and minimized. Only in the last few years, this is encouraged by the application of risk assessment, management, and communication. This work presents a probabilistic quantitative microbiological risk assessment and management of Listeria monocytogenes in ready-to-eat lettuce salads in Spain. For risk assessment, a guideline provided by Codex Alimentarius was followed. Food chain was modeled from processing of raw material at the factory up to consumption. Different assumptions were made to describe the variables of the model by probability distributions or mathematical models. Monte Carlo simulations of the model were run to estimate the number of cases in low-risk and high-risk populations. Although results deviated from the number of cases observed in Spain, given an ideal situation of 100% compliance of the microbiological criterion ≤100 cfu/g throughout the shelf-life of the product, the resulting number of cases was near the real situation. From the 4 risk management measures simulated, the injection of a mixture of gases into packages at manufacture (CO2 about 5.5%, O2 about 3%, and N2 for the balance) was the most effective in reducing the number of cases, followed by 4 d of storage at home and prevention of high-risk consumers from consumption of ready-to-eat lettuce salads. More research and cooperation between different stakeholder organizations are needed in order to progressively improve the model. With this work, a breakthrough has been made with regards to risk assessment and management procedures and implementation.