Meta-analysis of antimicrobial activity of Allium, Ocimum, and Thymus spp. confirms their promising application for increasing food safety.

Biopreservation strategies such as the use of Mediterranean plant extracts to ensure food safety are promising to deal with the emergence of antimicrobial resistances and the overreliance on food chemical additives. In the last few decades, antimicrobial susceptibility testing (AST) for evaluating the in vitro antibacterial potential of plant extracts against the most relevant foodborne pathogens has been widely reported in the literature. The current meta-analysis aimed to summarise and analyse the extensive evidence available in the literature regarding the in vitro antimicrobial capability of Allium, Ocimum and Thymus spp. extracts against foodborne pathogens. A systematic review was carried out to gather data on AST results of these extracts against Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Escherichia coli and Bacillus cereus, including inhibition diameters (ID) and minimum inhibitory concentrations (MIC). A total of 742 records were gathered from a raw collection of 2,065 articles. Weighted mixed-effect linear models were adjusted to data to obtain pooled ID, pooled MIC and the relationship between both model estimations and observations. The pooled results revealed B. cereus as the most susceptible bacteria to Allium sativum (pooled ID = 20.64 ± 0.61 mm) by diffusion methods and S. aureus (pooled MIC = 0.146 mg/mL) by dilution methods. Diffusion methods did not yield conclusive results for Ocimum spp. extracts; however, the lowest pooled MIC was obtained for S. aureus (0.263 mg/mL). Among the foodborne pathogens evaluated, B. cereus showed the highest sensitivity to Thymus spp. extracts by both diffusion and dilution methods (pooled ID = 28.90 ± 2.34 mm and MIC = 0.075 mg/mL). The methodology used for plant extraction was found to not significantly affect MIC values (p > 0.05). Overall, the antimicrobial effectiveness of the studied extracts against Gram-positive and Gram-negative bacteria was demonstrated. Finally, the robustness of the meta-regression model was confirmed, also revealing an inversely proportional correlation between the ID and MIC measurements (p < 0.0001). These results provide a robust scientific basis on the factors affecting the in vitro antimicrobial efficacy of extracts from Mediterranean plants. They also provide valuable information for stakeholders involved in their industrial application in food, including producers, regulatory agencies and consumers which demand green-labelled foods.

Detection, Control, Risk Assessment, and Prevention of Foodborne Microorganisms.

Despite significant efforts from government and industry, enteric foodborne diseases continue to pose a substantial public health challenge worldwide [...].

Optimal experimental design (OED) for the growth rate of microbial populations. Are they really more "optimal" than uniform designs?

Secondary growth models from predictive microbiology can describe how the growth rate of microbial populations varies with environmental conditions. Because these models are built based on time and resource consuming experiments, model-based Optimal Experimental Design (OED) can be of interest to reduce the experimental load. In this study, we identify optimal experimental designs for two common models (full Ratkowsky and Cardinal Parameters Model (CPM)) for a different number of experiments (10-30). Calculations are also done fixing one or more model parameters, observing that this decision strongly affects the layout of the OED. Using in silico experiments, we conclude that OEDs are more informative than conventional (equidistant) designs with the same number of experiments. However, OEDs cluster the experiments near the growth limits (Xmin and Xmax) resulting in impractical designs with aggregated experimental runs ~10 times longer than conventional designs. To mitigate this, we propose a novel optimality criterion (i.e., the objective function) that accounts for the aggregated time. The novel criterion provides a reduction in parameter uncertainty with respect to the conventional design, without an increase in the experimental load. These results underline that an OED is only based on information theory (Fisher information), so the results can be impractical when actual experimental limitations are considered. The study also emphasizes that most OED schemes identify where to measure, but do not give an indication on how many experiments should be made. In this sense, numerical simulations can estimate the parameter uncertainty that would be obtained for a particular experimental design (OED or not). These results and methodologies (available in Open Code) can guide the design of future experiments for the development of secondary growth models.

Inter-strain variability on the cardinal parameters (pH and Aw) of clinical and food isolates of Listeria monocytogenes using turbidimetric measurements.

Listeria monocytogenes is a foodborne pathogen which, in 2021, was considered the fifth most commonly reported zoonosis in humans in the European Union (EU). Ready-to-eat (RTE) fishery products, deli meats or soft cheeses have been mostly involved in food safety alerts and outbreaks in the last years. Hurdle technology by food industries has been widely used to enhance the safety of foods. Among the barriers, the application of acid and osmotic stress during processing is extensively used worldwide. This study aims to gain knowledge about the inter-strain variability of twenty-six clinical and food L. monocytogenes isolates with the estimation of their cardinal parameters using turbidimetric measurements. To analyse the data and to obtain the estimated cardinal values, a common statistical procedure was set up. The estimation of cardinal parameters showed a high inter-strain variability of L. monocytogenes, and no correlation was observed between Aw min and pHmin values for the studied strains. By grouping the strains in clinical, meat and fish origin, it was observed that strains from the meat group presented the lowest average pHmin values (4.57), thus showing potential acid adaptation. This work contributes to gain knowledge of the inter-strain variability of L. monocytogenes in relation with pH and Aw cardinal values, as well as provide a starting point for future validation studies in fish and meat food matrices.

Unveiling Fresh-Cut Lettuce Processing in Argentine Industries: Evaluating Salmonella Levels Using Predictive Microbiology Models.

A survey was performed to gather information on the processing steps, conditions, and practices employed by industries processing ready-to-eat (RTE) leafy vegetables in Argentina. A total of seven industries participated in the survey. A cluster analysis of the data obtained was performed to identify homogeneous groups among the participating industries. The data collected were used as inputs of two predictive microbiology models to estimate Salmonella concentrations after chlorine washing, during storage and distribution of final products, and to rank the different practices according to the final estimated Salmonella levels. Six different clusters were identified by evaluating the parameters, methods, and controls applied in each processing step, evidencing a great variability among industries. The disinfectant agent applied by all participating industries was sodium hypochlorite, though concentrations and application times differed among industries from 50 to 200 ppm for 30 to 110 s. Simulations using predictive models indicated that the reductions in Salmonella in RTE leafy vegetables would vary in the range of 1.70-2.95 log CFU/g during chlorine-washing depending on chlorine concentrations applied, washing times, and vegetable cutting size, which varied from 9 to 16 cm2 among industries. Moreover, Salmonella would be able to grow in RTE leafy vegetables during storage and distribution, achieving levels of up to 2 log CFU/g, considering the storage and transportation temperatures and times reported by the industries, which vary from 4 to 14 °C and from 18 to 30 h. These results could be used to prioritize risk-based sampling programs by Food Official Control or determine more adequate process parameters to mitigate Salmonella in RTE leafy vegetables. Additionally, the information gathered in this study is useful for microbiological risk assessments.

Dynamic Thermal Treatments in Green Coconut Water Induce Dynamic Stress Adaptation of Listeria innocua That Increases Its Thermal Resistance.

The global coconut water market is projected to grow in the upcoming years, attributed to its numerous health benefits. However, due to its susceptibility to microbial contamination and the limitations of non-thermal decontamination methods, thermal treatments remain the primary approach to ensure the shelf-life stability and the microbiological safety of the product. In this study, the thermal inactivation of Listeria innocua, a Listeria monocytogenes surrogate, was evaluated in coconut water and in tryptone soy broth (TSB) under both isothermal (50-60 °C) and dynamic conditions (from 30 to 60 °C, with temperature increases of 0.5, 1 and 5 °C/min). Mathematical models were used to analyse the inactivation data. The Geeraerd model effectively described the thermal inactivation of L. innocua in both TSB and coconut water under isothermal conditions, with close agreement between experimental data and model fits. Parameter estimates and analysis revealed that acidified TSB is a suitable surrogate medium for studying the thermal inactivation of L. innocua in coconut water, despite minor differences observed in the shoulder length of inactivation curves, likely attributed to the media composition. The models fitted to the data obtained at isothermal conditions fail to predict L. innocua responses under dynamic conditions. This is attributed to the stress acclimation phenomenon that takes place under dynamic conditions, where bacterial cells adapt to initial sub-lethal treatment stages, leading to increased thermal resistance. Fitting the Bigelow model directly to dynamic data with fixed z-values reveals a three-fold increase in D-values with lower heating rates, supporting the role of stress acclimation. The findings of this study aid in designing pasteurization treatments targeting L. innocua in coconut water and enable the establishment of safe, mild heat treatments for refrigerated, high-quality coconut water.

Variability in Physicochemical Parameters and Its Impact on Microbiological Quality and Occurrence of Foodborne Pathogens in Artisanal Italian Organic Salami.

Artisanal salami is produced in small-scale production plants, where the lack of full automation might result in higher variability in food intrinsic properties. The aim of the present study was to evaluate the inter- and intra-batch variability in physicochemical parameters and its impact on microbial quality and occurrence of foodborne pathogens on 480 samples collected from six batches of an artisanal Italian production of organic salami. Relatively high total bacterial counts (TBC) were found on the surface of the table in the stuffing room (4.29 ± 0.40 log cfu/cm2). High loads of Enterobacteriaceae in the meat mixture of batch 2 and TBC in batch 5 were associated with a higher occurrence of bacterial pathogens. During ripening, water activity (aw) and pH failed to reach values lower than 0.86 and 5.3, respectively. Six Staphylococcus aureus and four Listeria monocytogenes isolates were collected from the salami meat mixture during ripening and the processing environment. A total of 126 isolates of Enterobacteriaceae were characterized at a species level, with Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, and Citrobacter freundii isolated from the final products. Results suggest the relevance of first steps of production in terms of the hygiene of raw materials and handling during stuffing procedures, especially when the physicochemical parameters of the final products do not reach values that represent hurdles for foodborne pathogens.

In vitro antimicrobial activity of extracts and essential oils of Cinnamomum, Salvia, and Mentha spp. against foodborne pathogens: A meta-analysis study.

Essential oils (EOs) are a class of natural products that exhibit potent antimicrobial properties against a broad spectrum of bacteria. Inhibition diameters (IDs) and minimum inhibitory concentrations (MICs) are the typical measures of antimicrobial activity for extracts and EOs obtained from Cinnamomum, Salvia, and Mentha species. This study used a meta-analytical regression analysis to investigate the correlation between ID and MIC measurements and the variability in antimicrobial susceptibility tests. By utilizing pooled ID models, this study revealed significant differences in foodborne pathogens' susceptibility to extracts, which were dependent on both the plant species and the methodology employed (p < .05). Cassia showed the highest efficacy against Salmonella spp., exhibiting a pooled ID of 26.24 mm, while cinnamon demonstrated the highest efficacy against Bacillus cereus, with a pooled ID of 23.35 mm. Mint extract showed the greatest efficacy against Escherichia coli and Staphylococcus aureus. Interestingly, cinnamon extract demonstrated the lowest effect against Shiga toxin-producing E. coli, with a pooled ID of only 8.07 mm, whereas its EOs were the most effective against this bacterial strain. The study found that plant species influenced the MIC, while the methodology did not affect MIC measurements (p > .05). An inverse correlation between ID and MIC measurements was identified (p < .0001). These findings suggest that extracts and EOs obtained from Cinnamomum, Salvia, and Mentha spp. have the potential to inhibit bacterial growth. The study highlights the importance of considering various factors that may influence ID and MIC measurements when assessing the effectiveness of antimicrobial agents.

Assessing Listeria monocytogenes growth kinetics in rice pudding at different storage temperatures.

Rice pudding is a popular artisanal dairy dessert highly consumed in the main rice-producing countries, including Egypt. This study aimed to evaluate and model the growth of Listeria monocytogenes in rice pudding dessert stored at different temperatures (4-25 °C) over its shelf-life. Lab-scale rice pudding samples were prepared following a traditional Egyptian recipe and inoculated with a three-strain cocktail of L. monocytogenes (ca. 3 × 102 cfu/g). Inoculated rice pudding samples (pH = 6.67 ± 0.06 and aw = 0.99 ± 0.002) were stored at different isothermal conditions (4, 8, 12, 18, and 25 °C) and microbiologically analysed for up to 30 days for pathogen quantification by plate count methodology. Global regression analysis was used to fit the Baranyi model coupled with the Ratkowsky model to growth data, relating L. monocytogenes concentrations (N, log cfu/g) with storage temperature (°C) and times (d). Model validation was performed using published independent data. L. monocytogenes growth potential increased by increasing storage temperature. The estimated Ratkowsky model parameters were b = 0.0819 ± 0.0017 log cfu/d·°C and Tmin = -3.28 ± 0.20 °C. The indices RMSE = 0.39 and R2adj = 0.97 indicated a good agreement between the experimental data and the model predictions. The estimated maximum growth rate (µmax) values ranged between 0.355 and 5.363 log cfu/d from 4 to 25 °C. The model was successfully validated using published L. monocytogenes Scott A and California strains growth data in rice pudding samples stored at 5, 12 and 22 °C, as evidenced by the assessed statistical indices. The predictive model developed and validated in this study will aid in decision-making regarding the microbiological safety of rice pudding dessert with respect to L. monocytogenes growth, considering a wide range of storage temperatures.

Tracking microbial quality, safety and environmental contamination sources in artisanal goat cheesemaking factories.

A harmonised microbiological survey was performed in two artisanal factories of raw goat milk cheeses (A and B) located in the Andalusian region (Spain). A total of 165 different control points or samples (raw materials, final products, food-contact surfaces [FCS], and air) were examined as microbial and pathogen sources of contamination of artisanal goat raw milk cheeses. For raw milk samples analysed from both producers, the concentrations of aerobic-mesophilic bacteria (AMB), total coliforms, coagulase-positive Staphylococcus spp. (CPS), lactic-acid bacteria (LAB) and moulds and yeasts ranged between 3.48 and 8.59, 2.45-5.48, 3.42-4.81, 4.99-8.59 and 3.35-6.85 log cfu/mL respectively. For the same microbial groups, the analysis of raw milk cheeses revealed concentrations ranging from 7.82 to 8.88, 2.00-6.82, 2.00-5.28, 8.11-9.57 and 2.00-5.76 log cfu/g, respectively. Although the raw material analysed from producer A presented higher microbial loads and between-batch variability, it was B the producer with the most loaded final products. Regarding the microbial air quality, the fermentation area, storage room, milk reception and packaging room were the most AMB loaded places, while the ripening chamber was the area with higher fungal loads in bioareosol from both producers. Conveyor belts, cutting machine, storage boxes and brine tank were highlighted as the most contaminated FCS evaluated. Staphylococcus aureus was the only pathogen detected within the set of 51 isolates from samples as revealed by MALDI-TOF and molecular PCR, with a prevalence of 12.5% for samples from the producer B. The public health risk attributed to the consumption of artisanal goat cheese should not be neglected, and may consider the whole cheesemaking processing chain, from microbiological quality of raw milk to the ready-to-eat final product, especially concerning the presence of S. aureus.

Meta-Analysis of In Vitro Antimicrobial Capacity of Extracts and Essential Oils of Syzygium aromaticum, Citrus L. and Origanum L.: Contrasting the Results of Different Antimicrobial Susceptibility Methods.

Diffusion methods, including agar disk-diffusion and agar well-diffusion, as well as dilution methods such as broth and agar dilution, are frequently employed to evaluate the antimicrobial capacity of extracts and essential oils (EOs) derived from Origanum L., Syzygium aromaticum, and Citrus L. The results are reported as inhibition diameters (IDs) and minimum inhibitory concentrations (MICs), respectively. In order to investigate potential sources of variability in antimicrobial susceptibility testing results and to assess whether a correlation exists between ID and MIC measurements, meta-analytical regression models were built using in vitro data obtained through a systematic literature search. The pooled ID models revealed varied bacterial susceptibilities to the extracts and in some cases, the plant species and methodology utilised impacted the measurements obtained (p < 0.05). Lemon and orange extracts were found to be most effective against E. coli (24.4 ± 1.21 and 16.5 ± 0.84 mm, respectively), while oregano extracts exhibited the highest level of effectiveness against B. cereus (22.3 ± 1.73 mm). Clove extracts were observed to be most effective against B. cereus and demonstrated the general trend that the well-diffusion method tends to produce higher ID (20.5 ± 1.36 mm) than the disk-diffusion method (16.3 ± 1.40 mm). Although the plant species had an impact on MIC, there is no evidence to suggest that the methodology employed had an effect on MIC (p > 0.05). The ID-MIC model revealed an inverse correlation (R2 = 47.7%) and highlighted the fact that the extract dose highly modulated the relationship (p < 0.0001). The findings of this study encourage the use of extracts and EOs derived from Origanum, Syzygium aromaticum, and Citrus to prevent bacterial growth. Additionally, this study underscores several variables that can impact ID and MIC measurements and expose the correlation between the two types of results.

High pressure processing to control Salmonella in raw pet food without compromising the freshness appearance: The impact of acidulation and frozen storage.

The trend of feeding dogs and cats with raw pet food claiming health benefits poses health concerns due to the occurrence of pathogenic bacteria. High pressure processing (HPP) allows the non-thermal inactivation of microorganisms, preserving the nutritional characteristics with minimal impact on organoleptic traits of food. The present study aimed to evaluate and model the effect of HPP application (450-750 MPa for 0-7 min) on the inactivation of Salmonella, endogenous microbiota and colour of raw pet food formulated with different concentrations of lactic acid (0-7.2 g/kg) as natural antimicrobial. Additionally, the effect of a subsequent frozen storage of pressurized product was assessed. Salmonella inactivation ranged between 1 and 9 log, depending on the combination of conditions. According to the polynomial model obtained, the effect of pressure was linear, while a quadratic term was also included for holding time (depicting the occurrence of a resistant tail at ca. 4-6 min). The effect of lactic acid was dependent on the pressure level, being most relevant for treatments below 600 MPa. Frozen storage after HPP prevented the pathogen recovery and caused a further Salmonella inactivation enhanced by lactic acid in most of the treatments. Endogenous microbial groups were significantly reduced by HPP to below the detection level in several conditions. In general, little effect of HPP on the instrumental colour parameters was observed, except for a slight increase in lightness, which was hardly appreciable from visual observation. High pressure processing emerges as a relevant technology for the control of Salmonella spp. and to manage the microbiological safety of raw pet food. The mathematical model can be used as decision support tool to design safer raw pet food, while keeping the desired freshness appearance of the products.

Occurrence of foodborne pathogens in Italian soft artisanal cheeses displaying different intra- and inter-batch variability of physicochemical and microbiological parameters.

Artisanal cheeses are produced in small-scale production plants, where the lack of full automation and control of environmental and processing parameters suggests a potential risk of microbial contamination. The aim of this study was to perform a longitudinal survey in an Italian artisanal factory producing a spreadable soft cheese with no rind to evaluate the inter- and intra-batch variability of physicochemical and microbial parameters on a total of 720 environmental and cheese samples. Specifically on cheese samples, the evaluation was additionally performed on physicochemical parameters. Cheese samples were additionally collected during 15 days of storage at constant temperatures of 2 and 8°C, as well as a dynamic profile of 2°C for 5 days and 8°C for 10 days. Furthermore, Enterobacteriaceae isolates were identified at species level to have a better knowledge of the environmental and cheese microbiota potentially harboring human pathogens. High inter-batch variability was observed for lactic acid bacteria (LAB) and total bacteria count (TBC) in cheese at the end of production but not for pH and water activity. A temperature of 8°C was associated with a significantly higher load of Enterobacteriaceae in cheeses belonging to batch 6 at the end of storage, and this temperature also corresponded with the highest increase in LAB and TBC loads over cheese shelf life. Results from generalized linear mixed models (GLMMs) indicated that drains in the warm room and the packaging area were associated with higher levels of TBC and Enterobacteriaceae in cheese. Regarding foodborne pathogens, no sample was positive for verotoxigenic Escherichia coli (VTEC) or Listeria monocytogenes, whereas six Staphylococcus aureus and one Salmonella pullorum isolates were collected in cheese samples during storage and processing, respectively. Regarding Enterobacteriaceae, 166 isolates were identified at species level from all batches, with most isolates belonging to Klebsiella oxytoca and pneumoniae, Enterobacter cloacae, Hafnia alvei, and Citrobacter freundii evidencing the need to focus on standardizing the microbial quality of cow milk and on hygienic procedures for cleaning and disinfection especially in warm and maturation rooms. Further studies should be performed to investigate the potential pathogenicity and antimicrobial resistance of the identified Enterobacteriaceae species in artisanal cheeses.

Listeria monocytogenes survives better at lower storage temperatures in regular and low-salt soft and cured cheeses.

The behaviour of Listeria monocytogenes was investigated in soft pasteurized milk cheese elaborated with different salt concentrations (1.17 and 0.30% w/w) and in cured raw sheep milk cheese over storage up to 189 days at different isothermal conditions. Commercial 25-g cheese samples were inoculated with a 4-strain cocktail of L. monocytogenes (serovars 4b, 1/2a, 1/2b and 1/2c) at approximately 104 CFU/g. The inoculated samples were stored at 4 and 22 °C and withdrawn at proper intervals for L. monocytogenes enumeration. The prevalence of the different serovar strains of L. monocytogenes was characterized on soft cheese samples over storage at 4 °C using multiplex PCR. Salt reduction did not affect the survival of L. monocytogenes in soft cheeses and a maximum of 1-log reduction was observed in both regular and low-salt cheeses after 189 days of storage at 4 °C. The pathogen showed greater survival capacity in both soft and cured cheeses during storage at 4 °C compared to the storage at 22 °C, where more than 2.5 log reductions were computed. The fate of L. monocytogenes was described through a Weibull model fitted to survival data. The time required for a first tenfold reduction of the L. monocytogenes population (δ) at 4 °C is around 150 days in soft and 72 days in cured cheeses. At 22 °C, the estimated δ values are at least 60% lower in both cheese types. Among the four L. monocytogenes serovars present in the inoculated cocktail, the serovar 4b strain was the most sensitive to refrigerated storage, while the prevalence of serovar 1/2c strain increased over time in soft cheeses. Overall, the data obtained in this study help to deepen knowledge into factors affecting L. monocytogenes behaviour on cheeses and evidenced the variability between serovars in terms of survival capacity, which may be considered when performing microbial risk assessments.

Enhanced high hydrostatic pressure lethality in acidulated raw pet food formulations was pathogen species and strain dependent.

Feeding dogs and cats with raw meat-based pet food is taking relevance in the recent years. The high aw of these products together with the no cooking before its consumption by the animal pose a risk due to the potential occurrence and growth of foodborne pathogens. High pressure processing (HPP) is a non-thermal emerging technology that can be used as a lethality treatment to inactivate microorganisms with a minimum impact on the sensory and nutritional traits of the product. The purpose of the present study was to evaluate the variability in pressure resistance of different strains of the relevant foodborne pathogens Salmonella spp., Escherichia coli and Listeria monocytogenes in raw pet food formulated without and with lactic acid. In general, Salmonella and L.monocytogenes strains showed a higher resistance to HPP than E. coli strains. In lactic acid acidulated formulations, the susceptibility to HPP of L. monocytogenes was markedly enhanced. The resistance to HPP was not only dependent on the microorganism but also on the strain. Thus, the selection of the proper strains should be taken into account when designing and validating the application of HPP as a control measure within the HACCP plan.

Evaluating the fate of Escherichia coli O157:H7 and Salmonella spp. on cucumbers.

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.

From Cheese-Making to Consumption: Exploring the Microbial Safety of Cheeses through Predictive Microbiology Models.

Cheeses are traditional products widely consumed throughout the world that have been frequently implicated in foodborne outbreaks. Predictive microbiology models are relevant tools to estimate microbial behavior in these products. The objective of this study was to conduct a review on the available modeling approaches developed in cheeses, and to identify the main microbial targets of concern and the factors affecting microbial behavior in these products. Listeria monocytogenes has been identified as the main hazard evaluated in modelling studies. The pH, aw, lactic acid concentration and temperature have been the main factors contemplated as independent variables in models. Other aspects such as the use of raw or pasteurized milk, starter cultures, and factors inherent to the contaminating pathogen have also been evaluated. In general, depending on the production process, storage conditions, and physicochemical characteristics, microorganisms can grow or die-off in cheeses. The classical two-step modeling has been the most common approach performed to develop predictive models. Other modeling approaches, including microbial interaction, growth boundary, response surface methodology, and neural networks, have also been performed. Validated models have been integrated into user-friendly software tools to be used to obtain estimates of microbial behavior in a quick and easy manner. Future studies should investigate the fate of other target bacterial pathogens, such as spore-forming bacteria, and the dynamic character of the production process of cheeses, among other aspects. The information compiled in this study helps to deepen the knowledge on the predictive microbiology field in the context of cheese production and storage.

'MicroHibro': A software tool for predictive microbiology and microbial risk assessment in foods.

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.

Model for Listeria monocytogenes inactivation by high hydrostatic pressure processing in Spanish chorizo sausage.

A central composite design was implemented to study the effect of three factors on HHP-induced L. monocytogenes inactivation in Spanish chorizo sausage, in order to increase its effectiveness: product aw (0.79-0.92), pressure intensities (349-600 MPa, at 18 °C) and holding time (0-12.53 min). Response surface methodology was implemented with backward stepwise regression to generate a model that best fitted to the experimental data. All the three factors studied significantly influenced HHP inactivation of L. monocytogenes (p < 0.05). Pathogen reductions increased as the pressure and duration of HHP treatments rose. Low values of aw seemed to exert a protective effect on L. monocytogenes and pressures below 400 MPa did not lead to significant pathogen reductions. The model was validated with independent published data. Accuracy and bias factors were also determined to evaluate the performance of the developed model, which was considered acceptable for prediction purposes. The model generated represents a mathematical tool that will help food manufacturers to improve the efficacy of HHP processing of chorizo sausage and observe the regulatory authority's specifications regarding L. monocytogenes levels while maintaining food safety.

A Probabilistic Decision-Making Scoring System for Quality and Safety Management in Aloreña de Málaga Table Olive Processing.

Table olives are one of the most representatives and consumed fermented vegetables in Mediterranean countries. However, there is an evident lack of standardization of production processes and HACCP systems thus implying the need of establishing decision-making tools allowing their commercialization and shelf-life extension. The present work aims at developing a decision-making scoring system by means of a probabilistic assessment to standardize production process of Aloreña de Málaga table olives based on the identification of potential hazards or deficiencies in hygienic processes for the subsequent implementation of corrective measures. A total of 658 microbiological and physico-chemical data were collected over three consecutive olive campaigns (2014-2016) to measure the variability and relative importance of each elaboration step on total hygienic quality and product safety. Three representative companies were visited to collect samples from food-contact surfaces, olive fruits, brines, air environment, olive dressings, water tanks, and finished/packaged products. A probabilistic assessment was done based on the establishment of Performance Hygiene and Safety Scores (PHSS 0-100%) through a standardized system for evaluating product acceptability. The mean value of the global PHSS for the Aloreña de Málaga table olives processing (PHHSFTOT) was 64.82% (90th CI: 52.78-76.39%) indicating the high variability among facilities in the evaluated processing steps on final product quality and safety. Washing and cracking, and selection and addition of olive dressings were detected as the most deficient ones in relation to PHSSFi values (p < 0.05) (mean = 53.02 and 56.62%, respectively). The relative contribution of each processing step was quantified by different experts (n = 25) from the Aloreña de Málaga table olive sector through a weighted PHSS (PHSSw). The mean value of PHSSw was 65.53% (90th CI: 53.12-77.52%). The final processing steps obtained higher values for PHSSw being the finished product the most relevant one (mean = 18.44%; 90% CI: 10.34-25.33%). Sensitivity analysis concluded that intervention measures focused on reducing the contamination of washing brines could lead to an improvement of PHSSFTOT value to 67.03%. The present work can be potentially applied in the Aloreña de Málaga table olive food sector for improving food quality and safety assurance.