Search

In the present manuscript, we describe the mechanisms involved in the yeast-to-hypha dimorphic transition of the plant pathogenic Basidiomycota fungus Ustilago maydis. During its life cycle, U. maydis presents two stages: one in the form of haploid saprophytic yeasts that divide by budding and the other that is the product of the mating of sexually compatible yeast cells (sporidia), in the form of mycelial dikaryons that invade the plant host. The occurrence of the involved dimorphic transition is controlled by the two mating loci a and b. In addition, the dimorphic event can be obtained in vitro by different stimuli: change in the pH of the growth medium, use of different carbon sources, and by nitrogen depletion. The presence of other factors and mechanisms may affect this phenomenon; among these, we may cite the PKA and MAPK signal transduction pathways, polyamines, and factors that affect the structure of the nucleosomes. Some of these factors and conditions may affect all these dimorphic events, or they may be specific for only one or more but not all the processes involved. The conclusion reached by these experiments is that U. maydis has constituted a useful model for the analysis of the mechanisms involved in cell differentiation of fungi in general.

Biopreservation is a strategy that has been extensively covered by the scientific literature from a variety of perspectives. However, the development of quantitative modelling approaches has received little attention, despite the usefulness of these tools for the food industry to assess the performance and to set the optimal application conditions. The objective of this study was to evaluate and model the interaction between the antilisteria strain Latilactobacillus sakei CTC494 (sakacin K producer) and Listeria monocytogenes in vacuum-packaged sliced cooked ham. Cooked ham was sliced under aseptic conditions and inoculated with L. monocytogenes CTC1034 and/or L. sakei CTC494 in monoculture and coculture at 10:10, 10:103 and 10:105 cfu/g ratios of pathogen:bioprotective cultures. Samples were vacuum packaged and stored at isothermal temperature (2, 5, 10 and 15 °C). The growth of the two bacteria was monitored by plate counting. The Logistic growth model was applied to estimate the growth kinetic parameters (N0, λ, μmax, Nmax). The effect of storage temperature was modelled using the hyperbola (λ) and Ratkowsky (μmax) models. The simple Jameson-effect model, its modifications including the Ncri and the interaction γ factor, and the predator-prey Lotka Volterra model were used to characterize the interaction between both microorganisms. Two additional experiments at non-isothermal temperature conditions were also carried out to assess the predictive performance of the developed models through the Acceptable Simulation Zone (ASZ) approach. In monoculture conditions, L. monocytogenes and L. sakei CTC494 grew at all temperatures. In coculture conditions, L. sakei CTC494 had an inhibitory effect on L. monocytogenes by lowering the Nmax, especially with increasing levels of L. sakei CTC494 and lowering the storage temperature. At the lowest temperature (2 °C) L. sakei CTC494 was able to completely inhibit the growth of L. monocytogenes when added at a concentration 3 and 5 Log higher than that of the pathogen. The inhibitory effect of the L. sakei CTC494 against L. monocytogenes was properly characterized and modelled using the modified Jameson-effect with interaction γ factor model. The developed interaction model was tested under non-isothermal conditions, resulting in ASZ values ≥83%. This study shows the potential of L. sakei CTC494 in the biopreservation of vacuum-packaged cooked ham against L. monocytogenes. The developed interaction model can be useful for the industry as a risk management tool to assess and set biopreservation strategies for the control of L. monocytogenes in cooked ham. info:eu-repo/semantics/acceptedVersion

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 (

19 Páginas.-- 6 Figuras.-- 2 Tablas, Olea europaea L. leaves constitute a source of bioactive compounds with recognized benefits for both human health and technological purposes. In the present work, different extracts from olive leaves were obtained by the application of two extraction methods, Soxhlet and microwave-assisted extraction (MAE), and six solvents (distilled water, ethanolic and glycerol mixtures solvents). MAE was applied under 40, 60 and 80 °C for 3, 6.5 and 10 min. The effect of the extraction method, solvent and treatment factors (the latter in MAE) on the total phenol content (TPC), the antioxidant activity (AA) and the phenolic profile of the extracts were all evaluated. The extracts showed high values of TPC (up to 76.1 mg GAE/g DW) and AA (up to 78 mg TE/g DW), with oleuropein being the most predominant compound in all extracts. The Soxhlet extraction method exhibited better yields in TPC than in MAE, although both methods presented comparable AA values. The water MAE extract presented the strongest antimicrobial activity against five foodborne pathogens, with minimum inhibitory concentration (MIC) values ranging from 2.5 to 60 mg/mL. MAE water extract is proposed to be exploited in the food and nutraceutical industry in the frame of a sustainable economy., The authors are grateful to the Ministry of Economy and Competitiveness of Spain for funding this research through the project CTQ2016-78729-R, to the Andalusian research project PY18-616 (Plan for Research, Development and Innovation 2020), and to the Andalusian knowledge transfer project AT17-5927 co-funded by the European Regional Development Fund instrument (ERDF Andalusian Operative Programme 2014–2020).

The aim of this study was to model the growth and survival behaviour of Salmonella Reading and endogenous lactic acid bacteria on fresh pre-cut iceberg lettuce stored under modified atmosphere packaging for 10 days at different temperatures (4, 8 and 15 &deg, C). The Baranyi and Weibull models were satisfactorily fitted to describe microbial growth and survival behaviour, respectively. Results indicated that lactic acid bacteria (LAB) could grow at all storage temperatures, while S. Reading grew only at 15 &deg, C. Specific growth rate values (&mu, max) for LAB ranged between 0.080 and 0.168 h&minus, 1 corresponding to the temperatures 4 and 15 &deg, C while for S. Reading at 15 &deg, C, &mu, max = 0.056 h&minus, 1. This result was compared with published predictive microbiology models for other Salmonella serovars in leafy greens, revealing that predictions from specific models could be valid for such a temperature, provided they were developed specifically in lettuce regardless of the type of serovars inoculated. The parameter delta obtained from the Weibull model for the pathogen was found to be 16.03 and 18.81 for 4 and 8 &deg, C, respectively, indicating that the pathogen underwent larger reduction levels at lower temperatures (2.8 log10 decrease at 4 &deg, C). These data suggest that this Salmonella serovar is especially sensitive to low temperatures, under the assayed conditions, while showcasing that a correct refrigeration could be an effective measure to control microbial risk in commercial packaged lettuce. Finally, the microbiological data and models from this study will be useful to consider more specifically the behaviour of S. Reading during transport and storage of fresh-cut lettuce, elucidating the contribution of this serovar to the risk by Salmonella in leafy green products.

18 Páginas.-- 7 Tablas.-- 3 Figuras, The Clostridium sp. is a large group of spore-forming, facultative or strictly anaerobic, Gram-positive bacteria that can produce food poisoning. The table olive industry is demanding alternative formulations to respond to market demand for the reduction of acidity and salt contents in final products. while maintaining the appearance of freshness of fruits. In this work, logistic regression models for non-adapted and acid-adapted Clostridium sp. strains were developed in laboratory medium to study the influence of pH, NaCl (%) and time on the probability of germination of their spores. A Clostridium sporogenes cocktail was not able to germinate at pH < 5.0, although the adaptation of the strains produced an increase in the probability of germination at 5.0–5.5 pH levels and 6% NaCl concentration. At acidic pH values (5.0), the adapted strains germinated after 10 days of incubation, while those which were non-adapted required 15 days. At pH 5.75 and with 4% NaCl, germination of the adapted strains took place before 7 days, while several replicates of the non-adapted strains did not germinate after 42 days of storage. The model was validated in natural green olive brines with good results (>81.7% correct prediction cases). The information will be useful for the industry and administration to assess the safety risk in the formulation of new processing conditions in table olives and other fermented vegetables., The research leading to these results has received funding from the Junta de Andalucía Government through the PrediAlo project (AGR-7755: www.predialo.science.com.es); the TOBE project (RTI2018-100883-B-I00) and FEDER European funds. This work has also been performed by the Research Group AGR-170 (HIBRO) of the Research Andalusian Plan.

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 Rio, 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.

We have previously described that laboratory strains of Ustilago maydis, a fungal pathogen of maize and its ancestor teosinte, harbor an intracellular bacterium that enables the fungus to fix nitrogen. However, it is not clear whether other strains isolated from nature also harbor endosymbiotic bacteria, and whether these fix nitrogen for its host. In the present study, we isolated U. maydis strains from naturally infected maize. All the isolated strains harbored intracellular bacteria as determined by PCR amplification of the 16S rRNA gene, and some of them showed capacity to fix nitrogen. That these are truly bacterial endosymbionts were shown by the fact that, after thorough treatments with CuSO4 followed by serial incubations with antibiotics, the aforementioned bacterial gene was still amplified in treated fungi. In all, these data support the notion that U. maydis-bacterium endosymbiosis is a general phenomenon in this species.

Chicken breast meat is an important vehicle of pathogenic Escherichia coli, which raises public health concerns. The objective of this study was to evaluate and model the combined effects of caffeine and UV-C irradiation on E. coli levels on raw chicken breast fillets. Chicken breast samples were pre-treated with different concentrations of caffeine (0–20 mM/g), inoculated with E. coli (ca. 107-108 cfu/g), and submitted to UV-C irradiation at doses ranging from 0 to 15 J/cm2. E. coli inactivation increased by increasing both caffeine concentrations and UV-C doses. Reductions of more than 5-logs were observed in caffeine-free samples at UV-C doses higher than 12 J/cm2, while the pre-treatment of samples with caffeine at 20 mM/g resulted in undetectable levels of E. coli after UV-C treatments at doses higher than 6 J/cm2. The Weibull model coupled with a linear secondary model was suitable to fit the inactivation data (RMSE = 0.48, R2adj = 0.95) and can be used to predict E. coli inactivation as a function of caffeine concentrations and UV-C doses. Thus, the model generated in this study can be applied by food-business operators for decision-making and for process optimization. Overall, the results derived from this study revealed that caffeine application on chicken breast samples helps to reduce the UV-C doses required to reach desired levels of E. coli inactivation.

Background The application of High Hydrostatic Pressure (HHP) processing technology as a non-thermal pasteurization method has been extensively investigated over the last two decades. Listeria monocytogenes is a relevant target for food safety due to its ability to grow and/or survive in a wide range of environmental conditions and be present at hazardous levels in food commodities where lethal treatments have not been carried out, such as some ready-to-eat foods (RTE). Scope and approach This review presents a compilation of modelling studies describing pressure-induced inactivation of L. monocytogenes in foods. The influence of a series of factors, including technological parameters, food matrix composition and the physiological state of bacterial cells on inactivation levels is also discussed, as it should be clearly understood and evaluated in order to set and optimize HHP processing conditions. Key findings and conclusions The use of mathematical models to predict the inactivation and probability of recovery of L. monocytogenes in foods during HHP application and subsequent storage can help food processors and managers to comply with the current microbiological regulations established for RTE foods, as well as optimize processing conditions.

Consumers' demand for ready-to-eat (RTE) turkey meat is attributed to its convenience and healthy properties. However, as cooked meat product it is subjected to post-process contamination, thus allowing presence and growth of microbial pathogens, such as Salmonella spp.. The aim of this study was to include a natural antimicrobial, thyme essential oil (TEO), on RTE turkey products in order to evaluate its effectiveness throughout the shelf life. To do so, the effect of four different formulations of cooked RTE turkey products on Salmonella Enteritidis behaviour was investigated. Products' slices were surface inoculated with S. Enteritidis (ca. 4 to 5logcfu/g), subsequently stored at 10 and 25°C and microbiologically analysed during 18 and 12days, respectively. Predictive microbiology models fitted to count data were used to evaluate microbial behaviour. Results showed that S. Enteritidis behaviour on RTE turkey products slices during storage was strongly dependent on temperature. The pathogen was able to grow on slices at all tested conditions during storage at 25°C and no statistical differences were detected (p>0.05) between growth parameters. At 10°C, different behaviour patterns were observed. The application of TEO led to higher Salmonella inactivation rates on a product exempt of chemical preservatives. The addition of this novel antimicrobial on meat products or its incorporation on meat active packaging systems as a part of hurdle technology could increase RTE turkey products safety while satisfying the demand of more natural foods.

In honey bee research conducted in apiaries, a large amount of information is usually generated requiring a flexible database for storing and retrieving data. Here, we developed a generic database pseudocode, based on the abstraction of the apiary system, for data collected from the colonies through time. We thank J Chávez-Galarza for the fruitful discussions during the design of the database architecture. This research was funded through the 2013-2014 BiodivERsA/FACCE-JPI joint call for research proposals, with the national funders “Fundação para a Ciência e Tecnologia” (Portugal), “Agence Nationale de la Recherche” (France), and “Ministério de Economia y Competividade” (Spain). info:eu-repo/semantics/publishedVersion

Most of the foodborne microbial diseases are linked to foods of animal origin such as milk, meat, and poultry. Nowadays, the presence of multi-drug resistant (MDR) pathogens in foods is becoming an increasingly public health concern worldwide due to the overuse of antimicrobial drugs in animal feed. MDR pathogens can enter the food chain by posing a significant risk to both animals and consumers. MDR pathogens causing infections are untreatable due to their resistance to various antibiotics, primarily cephalosporin and carbapenems and to their extended-spectrum beta-lactamase (ESBL)-producing capability. In addition, foods of animal origin and food-related environments can be likely vehicles for spreading of multi-drug resistance genes, which accelerates the thriving of global antibiotic resistance. This paper reviews the role of foods of animal origin as a vehicle for MDR pathogens, stressing the contribution of food processes, environments, and storage conditions in dissemination and reduction of antimicrobial resistances (AMRs). Controlling the growth of MDR microorganisms and limiting the transmission/expression of AMR genes in food ecosystems could be an effective mitigation strategy, putting the focus on food processes as a part of the solution for AMR in foods. Bioprotective cultures are also a promising and environmentally friendly technology to reduce the incidence of MDR pathogens, though caution is taken as microbial starters and probiotics can also carry AMR. Finally, applying Whole Genome Sequencing (WGS) and predictive microbiology, within a Risk Assessment framework, is key to get insight into those mechanisms and conditions along the food chain favoring or reducing AMR.

Ustilago maydis is a basidiomycete fungus of the subphylum Ustilaginomycotina. U. maydis is a biotrophic phytopathogen that causes common smut in maize (Zea mays L) and its ancestor teozintle (Zea mays ssp. parviglumis and ssp. mexicana). The economical importance of U. maydis is not as significant as other smuts because it does not cause heavy losses in agriculture but is considered a classic fungal model for studying the mechanism of DNA recombination and mating, other important aspects of fungal development, and the molecular mechanisms of fungal pathogenesis. The life cycle of U. maydis involves one saprophytic phase of haploid yeast form (sporidia) and a hyphal virulent dikaryon formed by mating of two sexually compatible sporidia. This invades the host growing in the hyphal form and finally forms tumors full of diploid teliospores that germinate with the formation of a phragmobasidium and four basidiospores. This cycle has been analyzed at the molecular level. Importantly, it was found that U. maydis may be pathogenic under axenic conditions for plants unrelated to maize, but does not complete the sexual cycle. The dimorphic yeast-to-hypha transition occurs also in vitro induced by growth with fatty acids or acetate and at acidic pH, developing into multicellular individuals, and unexpectedly forms basidiocarps in vitro completing the sexual cycle with the formation of holobasidia instead of phragmobasidia, septal pores, and fibulae. Interestingly, laboratory and natural strains of U. maydis harbor a bacterial symbiont that fixes N2.

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. info:eu-repo/semantics/acceptedVersion

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 (

Nowadays, microorganisms with probiotic or antimicrobial properties are receiving major attention as alternative resources for food preservation. Lactic acid bacteria are able to synthetize compounds with antimicrobial activity against pathogenic and spoilage flora. Among them, Lactobacillus plantarum ATCC 8014 has exhibited this capacity, and further studies reveal that the microorganism is able to produce bacteriocins. An assessment of the growth of L. plantarum ATCC 8014 at different conditions becomes crucial to predict its development in foods. A response surface model of the growth rate of L. plantarum was built in this study as a function of temperature (4, 7, 10, 13, and 16°C), pH (5.5, 6.0, 6.5, 7.0, and 7.5), and sodium chloride (0, 1.5, 3.0, 4.5, and 6.0%) and sodium lactate (0, 1, 2, 3, and 4%) concentrations. All the factors were statistically significant at a confidence level of 90% (p<0.10). When temperature and pH increased, there was a corresponding increase in the growth rate, while a negative relationship was observed between NaCl and Na-lactate concentrations and the growth parameter. A mathematical validation was carried out with additional conditions, demonstrating an excellent performance of the model. The developed model could be useful for designing foods with L. plantarum ATCC 8014 added as a probiotic.

The aim of this study was to analyze the influence of milk collection frequency (24 h versus 48 h) and milking dayshift (morning and evening) on total mesophilic aerobic bacteria (MAB) and psychrotrophic bacteria (PSY) counts in raw milk samples. MAB counts were determined by flow cytometry (BactoScan) and PSY counts by the plate counting agar method. An univariate statistical analysis was performed to find out significant differences among the studied factors. Results obtained showed that collecting milk every 24 h was effective in reducing MAB and PSY counts by 32 and 18%, respectively, compared to 48 h milk collection. This positive impact allowed reducing up to 4°C the temperature of the heat treatment in the dairy industry, thus involving energy savings of 22%. Milking during the mornings showed a significant reduction of MAB counts in comparison to milking performed during the evenings (P<0.05). These results are highly useful for the improvement of milk quality through the optimization of collection and milking systems set at primary production.

The study determined total mercury (Hg) content of 10 wild edible mushroom species collected in southern Spain. Results indicated that the highest Hg level corresponded to Boletus aereus with 10.28 ± 2.92 mg/kg DW (dry weight), while the lowest Hg level was found in Terfezia arenaria with 0.09 ± 0.08 mg/kg DW. Regarding the anatomic parts of the mushrooms, caps showed significantly higher Hg concentrations than stems in B. aereus, Amanita caesarea and Macrolepiota procera. The percentage of contribution to the Provisional Tolerable Weekly Intake (PTWI) for Hg was also calculated. Based on consumption data in Spain (0.011 kg/person/week), %PTWI was calculated ranging between 0.06 and 3.5% for mean Hg levels. When a high level of consumption was assumed (0.100 kg/person/week), %PTWI ranged between 0.58 and 31%. Results suggest that wild edible mushrooms in southern Spain could contribute with high Hg levels to the Spanish diet. However, the lack of consumption data for wild edible mushrooms made it difficult to give more definitive conclusions; hence further studies including specific consumption data will be needed.

This work investigated the potential of NIR technology to be applied in the dairy industry for the detection of micro-organisms. To this end, two types of cow milk samples were studied, one in which only bacterial biomass was considered and the other in which bacteria were cultured and grown in milk for 24 h. The study was carried out using two micro-organisms Escherichia coli and Pseudomonas aeruginosa. Both types of samples with different counts of both micro-organisms were analysed by a NIR analyser in the range 10 000–4000 cm−1 based on transmittance measurements. Multivariate models indicated that a better discrimination between micro-organisms was attained in those milk samples in which micro-organisms have been grown.

Food safety standards in the European Union include microbiological criteria and targets in primary production. The current paper provides a strategy to elucidate risk-based metrics such a potential Food Safety Objective for Campylobacter used as benchmark to derive possible Performance Objectives for the pathogen in broiler carcasses tested after chilling. The Performance Objectives were developed using the EFSA data collected on broiler carcasses during the monitoring study performed in 2008 in the European Union according to the evaluation of the different risk factors included in the survey. The FSO for Campylobacter was set at −1.2 log10 cfu/g (∼6 cfu/100 g). The Campylobacter concentrations after chilling resulting in a final concentration equal or below this proposed FSO were suggested as possible POs. The results obtained indicated that batches originating from previously thinned flocks can be more at risk of being colonized with Campylobacter. In fact, the estimated mean concentrations of Campylobacter on carcasses were 1.05 and 2.38 log10 cfu/g for non thinned and thinned flocks, respectively. Further, the impact of high Campylobacter contamination on carcasses (>2.5 log10 cfu/g) was shown since for those carcasses a reduction in PO values higher than 1.5 log10 cfu/g is needed to meet the FSO. In contrast no significant differences for PO values estimated were found between slaughterhouses with different capacity and for carcasses tested at different times from collection. This study provides a validated methodology for the estimation of risk-based metrics based on a quantitative approach allowing food safety authorities to develop specific microbiological criteria.

The objective of this study was to evaluate the behavior of Salmonella Enteritidis and Salmonella Typhimurium, the two most important serovars of salmonellosis, during the fermentation of yogurt. The microorganisms were enumerated in milk throughout the fermentation process at three initial inoculum levels (3, 5 and 7 log CFU/mL). DMFit software was used in the fitting procedure of the data (IFR, Norwich, UK, Version 3.5). The data provided sigmoidal curves that were successfully displayed with the Baranyi model. The results showed that the initial inoculum level did not affect the growth for both pathogens; thus, the µmax values (maximum specific growth rate) did not significantly differ across all the contamination levels, ranging from 0.26 to 0.38 for S. Enteritidis and from 0.50 to 0.56 log CFU/g/h for S. Typhimurium (P > 0.05). However, the µmax values significantly differed between the two serovars (P

In food safety and public health risk evaluations, microbiological exposure assessment plays a central role as it provides an estimation of both the likelihood and the level of the microbial hazard in a specified consumer portion of food and takes microbial behaviour into account. While until now mostly phenotypic data have been used in exposure assessment, mechanistic cellular information, obtained using omics techniques, will enable the fine tuning of exposure assessments to move towards the next generation of microbiological risk assessment. In particular, metagenomics can help in characterizing the food and factory environment microbiota (endogenous microbiota and potentially pathogens) and the changes over time under the environmental conditions associated with processing, preservation and storage. The difficulty lies in moving up to a quantitative exposure assessment, because the development of models that enable the prediction of dynamics of pathogens in a complex food ecosystem is still in its infancy in the food safety domain. In addition, collecting and storing the environmental data (metadata) required to inform the models has not yet been organised at a large scale. In contrast, progress in biomarker identification and characterization has already opened the possibility of making qualitative or even quantitative connection between process and formulation conditions and microbial responses at the strain level. In term of modelling approaches, without changing radically the usual model structure, changes in model inputs are expected: instead of (or as well as) building models upon phenotypic characteristics such as for example minimal temperature where growth is expected, exposure assessment models could use biomarker response intensity as inputs. These new generations of strain-level models will bring an added value in predicting the variability in pathogen behaviour. Altogether, these insights based upon omics techniques will increase our (quantitative) knowledge on pathogenic strains and consequently will reduce our uncertainty; the exposure assessment of a specific combination of pathogen and food will be then more accurate. This progress will benefit the whole community of safety assessors and research scientists from academia, regulatory agencies and industry. info:eu-repo/semantics/publishedVersion

The present work aimed at studying the cross contamination of apples by Salmonella during the processing of commercial fresh apples and its survival capacity on apple at room temperature. For the first study, the typical process of fresh apples was simulated at laboratory scale in which an apple that was artificially contaminated by Salmonella at different concentration levels (8, 6 and 5 log cfu/apple) was introduced in one batch and processed including a simulated transport/washing step and drying step using sponges to simulate the porous material used in the industry. Results indicated that at 8 log cfu/apple, 50% fresh apples were contaminated after processing, with all analysed environmental samples being positive for the pathogen, consisting of washing water and sponges. However, at lower inoculum levels (5–6 log cfu/apple) no cross contamination was detected in apples, and only environmental samples showed contamination by Salmonella after processing including both water and sponges. Experiments on the survival of Salmonella on apple showed that the pathogen was capable to survive for 12 days, only showing a significant drop at the end of the experiment. Finally, two-class attribute sampling plans were assessed as tool to detect Salmonella in different contamination scenarios in fresh apple. This analysis indicated that with the highest inoculum level, a total of 16 apples would be needed to reach 95% of detecting Salmonella (i.e. lot rejection). In turn, when low levels were assessed (5–6 log cfu/apple), a large number of apples ( n = 1021) would have to be sampled to obtain the same confidence level (95%). If the environment is sampled (i.e. water and sponges), a lower number of samples would be needed to detect the pathogen. However, the feasibility of environmental sampling has not been assessed from a practical point of view. Overall, the results in this study evidenced that cross contamination by Salmonella might occur during processing of fresh apples and subsequently, the pathogen might survive for a noticeable period of time.

Microbial survival on inert surfaces should be considered as an important factor to understand and quantify bacterial transfer to foods (i.e. cross contamination). The present work studied the survival of Escherichia coli O157:H7 and Salmonella spp. on stainless steel surfaces with different soiling substrates based on leafy vegetable juices. Furthermore, predictive models were proposed to describe survival patterns shown by both pathogens in the different substrates. E. coli O157:H7 and Salmonella spp. could be recovered until 192 and 168 h, respectively from surfaces soiled with chard, red cabbage, iceberg lettuce and romaine lettuce. However, in spinach and parsley juice substrates, microorganisms were not detected after 48 h. This survival 10/5/2013; Time:1:31:57time was much lower than that obtained in saline solution (120 h), used to simulate clean surfaces. The reduction rates observed in a period of time of 2 and 24 h for E. coli O157:H7 depended on the type of substrate and ranged 1.03–2.27 and 0.14–0.27 log cfu/cm 2 h, respectively. For Salmonella spp. results showed slightly lower drops, with 0.89–1.62 and 0.15–0.25 log cfu/cm 2 h, for 2 and 24 h, respectively. The Weibull model and Weibull + tail model seemed to be the most suitable mathematical functions describing survival of both microorganisms in the different substrates (adj-R 2 = 0.94–0.99). Experiments were developed simulating environmental conditions given in fresh-cut vegetable factories. Therefore, results and models generated in this study could be applied in quantitative risk assessment studies to obtain a more accurate representation of cross contamination, in addition to enabling the assessment of control strategies of both pathogens in the fresh-cut vegetable industry.

Alternative berry crops belonging with the Rosaceae family currently have a great importance in the US and Canadian agribusiness, in this sense, the Mexican serviceberry (Malacomeles denticulata) is a fruit shrub mainly distributed in Mexico that has potential uses like horticultural crop; however, little information about its variability has been reported. That is why this research aims to determine the variability of M. denticulata by SSR markers. The apple SSR markers had a good transferability to Malacomeles (74 %); all transferable SSR are polymorphic and sixteen of them had high values of polymorphic information content, Nei’s expected heterozygosity (He), and Shannon’s information index (I), even the apple SSR loci were not previously proved in this species. Besides it was found high variability in the germplasm of central Mexico mainly within populations (72 %), where three different genetic pool were detected suggesting that a breeding program could be based in individual selection. That is why it is necessary select outstanding individual into population before to make hybridizations among these selected parents.

Summary Lactobacillus plantarum growth in a vacuum-packaged cooked meat product under different storage temperatures (4, 10 and 16 °C) and the relation between the microorganism growth and sensory quality were investigated. The Gompertz model was fitted to experimental counts of L. plantarum showing a good fitting to growth curves at different temperatures. A root-square secondary model and linear model were satisfactorily fitted to estimated growth rates () and lag times (), respectively. The sensory attributes (colour, flavour, taste, appearance) were also evaluated due to their importance to the global quality (Q). The sensory deterioration was detected several days after L. plantarum reached the stationary phase, that is, 59, 45 and 25 days for 4, 10 and 16 °C, respectively. According to results, sensory deterioration was related to time when microorganism reached late stationary phase, phenomenon known as ‘delayed change’.

The survival of Escherichia coli O157:H7 and Staphylococcus aureus on stainless steel surfaces with Saline Solution (SS), Tryptone Soy Broth (TSB) and meat purge was studied, and based on results, mathematical models describing survival of pathogens as a function of time were proposed. Results indicated that S. aureus was able to survive longer than E. coli O157:H7 in all substrates. The type of substrate had a greater impact on the survival of E. coli O157:H7. This microorganism only remained viable for 8 and 50 h (hours) on surfaces with SS and TSB, respectively while on meat purge, the microorganism could be recovered after 200 h. For S. aureus, SS and TSB led to similar survival times (250 h) whereas on meat purge, survival capacity increased to 800 h. Survival data for S. aureus could be well described by a log-linear model or a Weibull model depending on the type of substrate (R(2) > 0.85). E. coli O157:H7 displayed an evident tail zone which made the Weibull model more appropriate (R(2) > 0.94). These survival models may be used in quantitative risk assessment to produce more accurate risk estimates. Finally, the results highlight the importance of performing effective cleaning procedures to prevent cross contamination.

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.

Aim of the study: To introduce and describe FlorNExT®, a free cloud computing application to estimate growth and yield of maritime pine (Pinus pinaster Ait.) even-aged stands in the Northeast of Portugal (NE Portugal). Area of study: NE Portugal. Material and methods: FlorNExT® implements a dynamic growth and yield modelling framework which integrates transition functions for dominant height (site index curves) and basal area, as well as output functions for tree and stand volume, biomass, and carbon content. Main results: FlorNExT® is freely available from any device with an Internet connection at: http://flornext.esa.ipb.pt/. Research highlights: This application has been designed to make it possible for any stakeholder to easily estimate standing volume, biomass, and carbon content in maritime pine stands from stand data, as well as to estimate growth and yield based on four stand variables: age, density, dominant height, and basal area. FlorNExT® allows planning thinning treatments. FlorNExT® is a fundamental tool to support forest mobilization at local and regional scales in NE Portugal. Material and methods: FlorNExT® implements a dynamic growth and yield modelling framework which integrates transition functions for dominant height (site index curves) and basal area, as well as output functions for tree and stand volume, biomass, and carbon content. Main results: FlorNExT® is freely available from any device with an Internet connection at: http://flornext.esa.ipb.pt/. Research highlights: This application has been designed to make it possible for any stakeholder to easily estimate standing volume, biomass, and carbon content in maritime pine stands from stand data, as well as to estimate growth and yield based on four stand variables: age, density, dominant height, and basal area. FlorNExT® allows planning thinning treatments. FlorNExT® is a fundamental tool to support forest mobilization at local and regional scales in NE Portugal.

Chilled Ready-To-Eat (RTE) foods are of concern in hospital foodservices because they can support microbial growth when subjecting to time/temperature abuses during processing and distribution together with poor handling practices. This study was conducted in five different hospitals (A–E) of Southern Spain during 2008–2009 to perform an evaluation of their sanitary conditions and microbiological quality of two RTE meals: lettuce salads and cooked ham. A checklist based on hygiene principles embedded in Food European legislation was developed and applied in each hospital. In parallel, microbiological analysis of food contact surfaces, air quality and time/temperature measurements along the distribution chain were carried out. RTE samples (n = 150) were examined for mesophilic aerobic bacteria (MAB), total coliforms, coagulase-positive Staphylococci (CPS), Escherichia coli, Listeria spp. and Salmonella spp. Differences were found between hospitals regarding handling practices and cleanliness of working surfaces. Cooked ham samples presented lower counts of MAB and total coliforms (

Food industry is increasingly concerned in developing and applying rapid and nondestructive methods to offer safer and high quality foods to consumers. During the last years, Fourier transform near-infrared (FT-NIR) has been widely used to determine food quality based on spectrum. Likewise, FT-NIR has been proposed as an innovative and promising nondestructive rapid method capable to detect and identify microorganisms in foods; however, little progress has been made to date in this field. This study is a new attempt to apply FT-NIR technology to identify and quantify bacteria species in water-based systems in order to simulate water-based food matrices. For that, three different lactic acid bacteria—Lactobacillus plantarum, Leuconostoc mesenteroides, and Lactobacillus sakei—associated with spoilage in ready-to-eat meat, were analyzed by reflectance–transmitance FT-NIR in the spectral range 1,100–2,500 nm. Principal component analysis (PCA), and partial least squares (PLS) were applied to obtain prediction models. PCA and PLS showed a clear discrimination between the tested bacteria species whereas PLS method could succesfully quantify the concentration levels (3–9 log cfu/mL) and also distinguish between spoilage (7–9 log cfu/mL) and nonspoilage concentration levels (3–6 log cfu/mL). Results suggest that FT-NIR could be used efficiently to detect and quantify microorgasnisms in water-based food matrices. However, this study is an initial approach and therefore, it will be necessary to further research in order to really carry out its application to more complex food matrices and other microorganisms (i.e., food-borne pathogens).

Cooked meat ready-to-eat products are recognized to be contaminated during slicing which, in the last years, has been associated with several outbreaks. This work aimed to find out possible relation between the hygiene practice taking place at retail point during slicing of cooked meat products in small and medium-sized establishments (SMEs) and large-sized establishments (LEs) and the microbiological quality of sliced cooked meat products. For that, a checklist was drawn up and filled in based on scoring handling practice during slicing in different establishments in Cordoba (Southern Spain). In addition, sliced cooked meats were analyzed for different microbiological indicators and investigated for the presence of Listeria spp. and Listeria monocytogenes. Results indicated that SMEs showed a more deficient handling practices compared to LEs. In spite of these differences, microbiological counts indicated similar microbiological quality in cooked meat samples for both types of establishments. On the other hand, Listeria monocytogenes and Listeria inocua were isolated from 7.35% (5/68) and 8.82% (6/68) of analyzed samples, respectively. Positive samples for Listeria spp. were found in establishments which showed acceptable hygiene levels, though contamination could be associated to the lack of exclusiveness of slicers at retail points. Moreover, Listeria spp presence could not be statistically linked to any microbiological parameters; however, it was observed that seasonality influenced significantly (P

Listeria monocytogenes, when present, could be able to grow on Ready-to-Eat (RTE) iceberg lettuce. This investigation demonstrates that L. monocytogenes grows on packaged shredded iceberg lettuce held at 13 or 5 °C (maximum growth rate of 0.019 and 0.013 log cfu/h, respectively), with increments of 4.85 and 2.66 log cfu/g, respectively, after 14 days. No lag time was observed at 13 °C, in contrast with that at 5 °C (5.6 days). Initial atmosphere inside the packages was 4.65–6.2% CO2, 2.1–4.3% O2 and a balance of N2. Five predictive models taken from literature gave conservative predictions at 13 °C, although at 5 °C they were closer. Lactic acid and psychrotrophic bacteria showed similar growth at both temperatures. In general, the MAP system reported here retarded the growth of L. monocytogenes in comparison with other works without initial flushing of gases.

Bacterial transfer events are considered to be an important cause of transmission of food-borne diseases. In this review the most common ways of bacterial transfer to food are studied, concluding that high levels of moisture, contact time and pressure could result in higher transfer between surfaces. From a risk management approach, this means that the identification and elimination of certain tasks or activities involving higher levels of these factors could be helpful to prevent bacterial transfer. Besides, mathematical bacterial transfer models from literature are analysed, highlighting the use of transfer rates distributions to model bacterial transfer in a probabilistic Quantitative Microbiological Risk Assessment (QMRA) framework.

The risk assessment study of Listeria monocytogenes in ready-to-eat foods conducted by the U.S. Food and Drug Administration is an example of an extensive quantitative microbiological risk assessment that could be used by risk analysts and other scientists to obtain information and by managers and stakeholders to make decisions on food safety management. The present study was conducted to investigate how detailed sensitivity analysis can be used by assessors to extract more information on risk factors and how results can be communicated to managers and stakeholders in an understandable way. The extended sensitivity analysis revealed that the extremes at the right side of the dose distribution (at consumption, 9 to 11.5 log CFU per serving) were responsible for most of the cases of listeriosis simulated. For concentration at retail, values below the detection limit of 0.04 CFU/g and the often used limit for L. monocytogenes of 100 CFU/g (also at retail) were associated with a high number of annual cases of listeriosis (about 29 and 82%, respectively). This association can be explained by growth of L. monocytogenes at both average and extreme values of temperature and time, indicating that a wide distribution can lead to high risk levels. Another finding is the importance of the maximal population density (i.e., the maximum concentration of L. monocytogenes assumed at a certain temperature) for accurately estimating the risk of infection by opportunistic pathogens such as L. monocytogenes. According to the obtained results, mainly concentrations corresponding to the highest maximal population densities caused risk in the simulation. However, sensitivity analysis applied to the uncertainty parameters revealed that prevalence at retail was the most important source of uncertainty in the model.

Aim of study: To present the software utilities and explain how to use EucaTool®, a free cloud computing application developed to estimate the growth and production of seedling and clonal blue gum (Eucalyptus globulus Labill.) plantations in Galicia (NW Spain).Area of study: Galicia (NW Spain).Material and methods: EucaTool® implements a dynamic growth and production model that is valid for clonal and non-clonal blue gum plantations in the region. The model integrates transition functions for dominant height (site index curves), number of stems per hectare (mortality function) and basal area, as well as output functions for tree and stand volume, biomass and carbon content.Main results: EucaTool® can be freely accessed from any device with an Internet connection, from http://app.eucatool.com. In addition, useful information about the application is published on a related website: http://www.eucatool.com.Research highlights: The application has been designed to enable forest stakeholders to estimate volume, biomass and carbon content of forest plantations from individual trees, diameter classes or stand data, as well as to estimate growth and future production (indicating the optimal rotation age for maximum income) by measurement of only four stand variables: age, number of trees per hectare, dominant height and basal area.Keywords: forest management; biomass; seedling; clones; blue gum; forest tool.

The objective of this work was to study the growth potential of Escherichia coli O157:H7 and Salmonella spp. in leafy vegetable extracts at different temperature conditions.Cocktails of five strains of E. coli O157:H7 and of Salmonella enterica were used. Inoculated aqueous vegetable extracts were incubated at 8, 10, 16 and 20°C during 21 days. Microbial growth was monitored using Bioscreen C(®) . In spinach extract, results showed that for E. coli O157:H7 and Salmonella significant differences (P 0·05) for μabs (maximum absorbance rate) were obtained. For both pathogens, growth in chard was slightly lower. In contrast, iceberg lettuce and parsley showed the lowest values of μabs , below 0·008 h(-1) . The coefficients of variance (CoV) calculated for the different replicates evidenced that at low temperature (8°C) a more variable behaviour of both pathogens is expected (CoV 180%).This study provides evidence that aqueous extracts from vegetable tissues can result in distinct growth niche producing different response in various types of vegetables.Finally, these results can be used as basis to establish risk rankings of pathogens and leafy vegetable matrices with relation to their potential growth.

Changes in sensory quality and proliferation of pathogenic and spoilage microorganisms in pasteurized white asparagus were measured as a function of storage temperature. The main sensorial attributes studied with regard to storage time were the general appearance of the product, turgidity and package swelling. There was a significant correlation between the deterioration in sensorial quality and microbial evolution, which was even more marked at higher temperatures. After 13 days at 4°C, 6 days at 10°C, 5 days at 20°C and 84 h at 30°C organoleptic deterioration and, consequently, the end of shelf life were detected. A significant growth of spoilage flora was observed, which might be responsible for the drop in pH levels during the final period of analysis (from 6.3 to 3.5–4 until 148 h). There was greater lactic acid bacteria (LAB) growth as the temperature and CO2 concentration increased, reaching a final concentration of approximately 108 CFU g−1 at all temperatures. We found no positive samples of Clostridium spp at 10 and 4°C. Among the species isolated at 20–30°C Clostridium septicum and in lower proportions Clostridium perfringens were encountered. The possible presence of Listeria monocytogenes was analyzed, but there were no positive samples in the final product. Pasteurized white asparagus maintains good organoleptic characteristics with a relatively long shelf-life at low temperatures, which leads to an adequate acceptance among consumers. Copyright © 2006 Society of Chemical Industry

Prevalence and concentration of Listeria monocytogenes, Salmonella spp. and enteric pathogenic viruses (namely Hepatitis A-HAV, and noroviruses genogroup I-NoVGI and genogroup II-NoVGII) were determined in raw and RTE lettuce from a Spanish processing premise. Fifteen samplings were made from September 2010 to February 2012 (n=600 samples). Sampling strategies for pathogen detection were suggested by the characterization of the uncertainty in prevalence associated with the performance of two-class attributes sampling plans (c=0). A probabilistic model was run (1000 iterations) using a Bayesian approach with a conjugate beta distribution considering the impact of taking different number of samples on the proportion of positive samples and lots (within- and between-lot prevalence). No enumeration results were obtained for the pathogens tested. Presence of L. monocytogenes and NoVGII in RTE lettuce (10%) and NoVGI and NoVGII in unprocessed lettuce (10%) was obtained in the tested lots during cold season. Results evidenced that, as the number of samples increased, the probability of rejecting a contaminated lot became higher, yielding right-skewed distributions with values close to 1. According to our results, 25 samples would result in 80% of rejected lots, while 95% confidence level would be reached with n>100. However, although those levels would imply a unrealistic high number of samples making the application of the sampling plan unfeasible, these results might be useful for food operators and risk managers to know the underlying distributions of microbial contamination together with potential control measures to be applied to assure a safer production of minimally processed vegetables.

Fecal pathogen distributions in fresh-cut leafy vegetables are essential to develop suitable sampling plans so as to detect pathogen contaminations. In this study, a typical fresh-cut lettuce process was reproduced at pilot scale with different initial inoculum levels of Salmonella on lettuce (6–7, 4 and 1 log CFU/g). The pathogen was determined in all processed lettuce samples (n ≥ 50) and obtained count data were used to fit different probability distributions. The study showed that Salmonella is homogenously distributed on fresh-cut leafy vegetables as a result of processing (mainly washing) at all contamination levels. Negative binomial and Poisson-lognormal distributions were suitable to describe pathogen distribution at the high and medium levels. coefficient of variation modified ( CV *) indicated no overdispersion (i.e. clustering). Nevertheless, further research will be needed to assess the effect of using disinfectants in washing water on the final distribution pattern of pathogens in processed fresh-cut leafy vegetables.

The presence of heavy metals in big game meat may pose a risk to human health. The main objective of this paper is to carry out a risk assessment study (using a probabilistic and point-estimate approach) of lead intake by consumption of red deer and wild boar meat in Southern Spain based on Spanish data collected in the period 2003-2006. In general, the concentration levels found for wild boar meat (mean = 1291 µg kg(-1)) were much higher than those observed in red deer meat (mean = 326 µg kg(-1)). The results from a point-estimate risk assessment showed that the estimated average intake of lead among different exposure scenarios varied from 0.1 to 6.5 and from 0.3 to 38 µg kg(-1) week(-1) for red deer and wild boar meat, respectively; and from 0.3 to 35 µg kg(-1) week(-1) for individuals consuming both red deer and wild boar meat, and that the estimated intake of lead by consumption of big game meat differed significantly between hunters and non-hunters, it being higher for hunters. Besides this, results from the probabilistic risk assessment study corroborated the fact that risk is greater in hunter populations, reaching a maximum in individuals consuming only wild boar and both types of meat, with 0.4% and 0.2% of the population above the provisional tolerable weekly intake (PTWI), respectively. Likewise, the hunter populations consuming wild boar and both types of big game meat (red deer and wild boar meat) were exposed to the maximum lead level (56 µg kg(-1) week(-1)), which corresponded approximately to 224% of the PTWI. Further data and studies will be needed to give a complete risk estimation in which it will be crucial to consider the contribution to the lead intake level of other foods in the diet of both population groups.

The presence of Escherichia coli in contaminated food products is commonly attributed to faecal contamination when they are improperly handled and/or when inactivation treatments fail. Adaptation of E. coli at low pH and a w levels can vary at different temperatures depending on the serotype, thus more detailed studies are needed. In this work, a screening to assess the growth of four pathogenic serotypes of E. coli (O55:H6; O59:H21; O158:H23 and O157:H7) was performed. Subsequently, boundary models were elaborated with the fastest serotype selected at different temperatures (8, 12 and 16 °C), and inoculum levels (2, 3 and 4 log cfu/mL) as function of pH (7.00–5.00) and a w (0.999–0.960). Finally, the growth kinetics of E. coli was described in the conditions that allowed growth. Results obtained showed that the serotypes O157:H7 and O59:H21 did not grow at more stringent conditions (8 °C; pH 5.50), while the E. coli O158:H23 was the best adapted, resulting in faster growth. The logistic regression models presented a good adjustment to data observed since more than 96.7% of cases were correctly classified. The growth interface was shifted to more limited conditions as the inoculum size was higher. Detection times ( t d , h) and their variability were higher at low levels of the environmental factors studied. This work provides insight on the growth kinetics of E. coli at various environmental conditions.

The microbial behaviour of five enterotoxigenic strains of Staphylococcus aureus was studied in the growth/no growth domain. A polynomial logistic regression equation was fitted using a stepwise method to study the interaction of temperature (8, 10, 13, 16 and 19 °C), pH (4.5; 5.0; 5.5; 6.0; 6.5 7.0 and 7.5) and water activity (Aw) (19 levels ranging from 0.867 to 0.999) on the probability of growth. Out of the 284 conditions tested, 146 were chosen for model data and 138 intermediate conditions for validation data. A growth/no growth transition was obtained by increasing the number of replicates per condition (n = 30) in comparison to other published studies. The logistic regression model showed a good performance since 96.6% (141 out of 146 conditions) of the conditions for model data and 92.0% (127 out of 138 conditions) for validation data were correctly classified. The predictions indicated an abrupt growth/no growth interfaces occurred at low levels of temperature, pH and Aw. At 8 °C, S. aureus grew only at optimum levels of pH and Aw while at temperatures above 13 °C, growth of S. aureus was observed at pH = 4.5 and Aw = 0.96 (13 °C), 0.941 (16 °C) and 0.915 (19 °C). The optimal pH at which growth of S. aureus was detected earlier was 6.5. However, a slight decrease of the probability of growth was noticed in the pH interval of 7.0–7.5 at more stringent conditions. The ability of S. aureus to grow at low Aw was shown since growth was detected at Aw = 0.867 (T = 19 °C; pH = 7.0). Finally, a comparison of model predictions with literature data on growth/no growth responses of S. aureus in culture media and cooked meat was made. Model predictions agreed with published data in 94% of growth cases and in 62% of no growth cases. The latter discordance is highly associated to other environmental factors (such as other preservatives, strains etc.) included in published models that did not match the ones included in our study. This study can help manufacturers in making decision on the most appropriate formulations for food products in order to prevent S. aureus growth and enterotoxin production along their shelf-life., This work was partly financed by MICINN AGL2008-03298/ALI, the Excellence Project AGR-01879 (Junta de Andalucía) and by European ERDF funding.

Cross contamination is one of the most important contributing factors in foodborne illnesses originating in household environments. The objective of this research was to determine the transfer coefficients between a contaminated domestic slicing machine and a cooked meat product, during slicing. The microorganisms tested were Staphylococcus aureus (Gram positive) and Escherichia coli O157:H7 (Gram negative). The results showed that both microorganisms were able to transfer to all slices examined (20 successively sliced) and at different inoculum levels on the blade (10 8 ,1 0 6 and 10 4 cfu/blade). The results also showed that the number of log cfu transferred per slice, during slicing, decreased logarithmically for both microorganisms at inoculum levels of 8 and 6 log cfu/blade. The type of microorganism significantly influenced transfer coefficients (p < 0.05) and there was an interaction between inoculum level and transfer coefficient for S. aureus (p < 0.05), but not E. coli O157:H7. Finally, to describe bacterial transfer during slicing, two models (log-linear and Weibull) were fitted to concentration on slice data for both microorganisms (at 6 and 8 log cfu/blade), obtaining a good fit to data (R 2 P 0.73).