Modelling the low temperature growth boundaries of Salmonella Enteritidis in raw and pasteurized egg yolk, egg white and liquid whole egg: Influence of the initial concentration.

Salmonella is the most frequently reported cause of foodborne outbreaks with known origin in Europe, with eggs and egg products standing out as the most frequent food source (when it was known). The growth and survival of Salmonella in eggs and egg products have been extensively studied and, recently, it has been reported that factors such as the initial concentration and thermal history of the egg product can also influence its growth capability. Therefore, the objective of this study was to define the boundary zones of the growth/no growth domain of Salmonella Enteritidis (4 strains) as a function of temperature (low temperature boundary) and the initial concentration in different egg products. A series of polynomial logistic regression equations were successfully adjusted, allowing the study of these factors and their interaction on the probability of growth of S. Enteritidis in these products. Results obtained indicate that the minimum growth temperatures of Salmonella Enteritidis are higher in egg white (9.5-18.3 °C) than in egg yolk (7.1-7.8 °C) or liquid whole egg (7.2-7.9 °C). Results also demonstrate that in raw liquid whole egg and raw and pasteurized egg white, the minimum growth temperature of Salmonella Enteritidis does depend on the initial concentration. Similarly, the previous thermal history of the egg product only influenced the minimum growth temperature in some of them. On the other hand, large differences in the minimum growth temperatures among strains were observed in some products (up to approx. 6 °C in egg white). Finally, it should be noted that none of the strains grew at 5 °C under any of the conditions assayed. Therefore, storage of egg products (particularly whole liquid egg and egg yolk) below this temperature might be regarded/proposed as a good management approach. Our experimental approach has allowed us to provide a more accurate prediction of S. Enteritidis minimum growth temperatures in egg products by taking into account additional factors (initial concentration and thermal history) while also providing a quantification of the intra-specie variability. This would be of high relevance for improving the safety of egg products.

Effect of the Application of Ultrasound to Homogenize Milk and the Subsequent Pasteurization by Pulsed Electric Field, High Hydrostatic Pressure, and Microwaves.

The efficacy of applying ultrasound (US) as a system to homogenize emulsions has been widely demonstrated. However, research has not yet shown whether the effect achieved by homogenizing milk with US is modified by subsequent pasteurization treatments that use new processing technologies such as pulsed electric fields (PEF), microwaves (MW), and high hydrostatic pressure (HPP). The aim of this study was, therefore, to optimize the application of US for milk homogenization and to evaluate the effect of PEF, HPP, and MW pasteurization treatments on the sensorial, rheological, and microbiological properties of milk throughout its shelf life. To homogenize whole milk, a continuous US system (20 kHz, 0.204 kJ/mL, 100%, 40 °C) was used, and different ultrasonic intensities (0.25, 0.5, and 1.0 kJ/mL) were evaluated. The optimal ultrasonic treatment was selected on the basis of fat globule size distribution and pasteurization treatments by MW (5800 W, 1.8 L/min), PEF (120 kJ/kg, 20 kV/cm) and HPP (600 MPa, 2 min, 10 °C) was applied. The ultrasound intensity that achieved the highest reduction in fat globule size (0.22 ± 0.02 µm) and the most homogeneous distribution was 1.0 kJ/mL. Fat globule size was smaller than in commercial milk (82% of volume < 0.5 µm for US milk versus 97% of volume < 1.2 µm for commercial milk). That size was maintained after the application of the different pasteurization treatments, and the resulting milk had better emulsion stability than commercial milk. After 28 days of storage, no differences in viscosity (4.4-4.9 mPa s) were observed. HPP pasteurization had the greatest impact on color, leading to higher yellowness values than commercial milk. Microbial counts did not vary significantly after 28 days of storage, with counts below 102 CFU/mL for samples incubated at 15 °C and at 37 °C. In summary, the homogenization of milk obtained by US was not affected by subsequent pasteurization processes, regardless of the technology applied (MW, PEF, or HPP). Further research is needed to evaluate these procedures' effect on milk's nutritional and functional properties.

Evaluation of Pulsed Electric Fields (PEF) Parameters in the Inactivation of Anisakis Larvae in Saline Solution and Hake Meat.

Larvae of the nematode family Anisakidae are capable of causing parasitic infections in humans associated with the consumption of fishery products, leading to intestinal syndromes and allergic reactions. Anisakidae larvae are widely distributed geographically, with rates of parasitism close to 100% in certain fish species. Methods need to be established for their inactivation and elimination, especially in fishery products that are to be consumed raw, pickled, or salted, or which have been insufficiently treated to kill the parasite. Many strategies are currently available (such as freezing and heat treatment), but further ones, such as pulsed electric fields (PEF), have hardly been investigated until now. This study focuses on the experimental evaluation of the efficacy of PEF in the inactivation of Anisakis spp. larvae in terms of electric field strength, specific energy, and pulse width, as well as on the evaluation of the quality of fish samples after PEF treatment. Results show that viability of Anisakis was highly dependent on field strength and specific energy. Pulse width exerted a considerable influence at the lowest field strengths tested (1 kV/cm). Central composite design helped to define a PEF treatment of 3 kV/cm and 50 kJ/kg as the one capable of inactivating almost 100% of Anisakis present in pieces of hake, while affecting the investigated quality parameters (moisture, water holding capacity, and cooking loss) to a lesser extent than freezing and thawing. These results show that PEF could serve as an alternative to traditional freezing processes for the inactivation of Anisakis in fish.

Relationship between iron bioavailability and Salmonella Typhimurium fitness in raw and pasteurized liquid whole egg.

SALMONELLA: Enteritidis growth rates in liquid whole egg have been shown to be dependent on the initial inoculum dose and on the egg product's thermal history. This study's objective is to obtain further insight into the mechanisms underlying both phenomena. First we verified that Salmonella Typhimurium ATCC 14028s cells displayed the behavior already described for S. Enteritidis cells. Then, we carried out supplementation assays by adding different concentrations of egg-white antimicrobial proteins, iron, or siderophores to the egg samples (raw or pasteurized liquid whole egg, depending on the assay). These experiments revealed that addition of lysozyme (at the concentration at which it is present in liquid whole egg) did not affect Salmonella growth in pasteurized liquid whole egg, but that ovotransferrin as well as Ex-FABP caused a significant (p < 0.05) reduction in Salmonella growth rates in whole egg pasteurized at 70 °C for 1.5 min. Furthermore, we observed that the inactivation of ovotransferrin was dependent on treatment intensity within the range studied. On the other hand, addition of iron or siderophores to raw or low temperature (60 °C/3.5 min) pasteurized liquid whole egg increased the growth rate of Salmonella cells inoculated at a low initial dose. The concentration of these supplements required to reach the growth rate of cells inoculated at a high dose was lower for pasteurized than for raw egg. Finally, growth of a set of deletion mutants in genes coding for proteins related to different iron uptake systems, along with supplementation assays using spent medium revealed the key role of salmochelin in growth of S. Typhimurium in raw whole egg. In summary, our results strongly suggest that iron bioavailability determines the fitness (growth rates) of Salmonella cells in liquid whole egg. Thus, the higher the intensity of the thermal treatment applied to liquid egg, the more iron would be available, a phenomenon that would be linked to the denaturation of iron and/or siderophore binding egg proteins. Further work is still required to fully elucidate why lower Salmonella initial doses lead to lower growth rates, but it can be hypothesized that this might be related to a lower amount of siderophores being released to the medium (especially salmochelin), which would also limit iron bioavailability.

Relationship between growth ability, virulence, and resistance to food-processing related stresses in non-typhoidal Salmonellae.

The ability of Salmonella to resist and adapt to harsh conditions is one of the major features that have made this microorganism such a relevant health hazard. However, the impact of these resistance responses on other aspects of Salmonella physiology, such as virulence and growth ability, is still not fully understood. The objective of this study was to determine the maximum growth rates (in three different media), virulence (adhesion and invasion of Caco-2 cells), and other phenotypic characteristics (biofilm-forming ability and antimicrobial resistance) of 23 Salmonella strains belonging to different serovars, and to compare them with their previously determined stress resistance parameters. Significant differences (p < 0.05) in growth rates, virulence, and biofilm-forming ability were found among the 23 strains studied. Nevertheless, whereas less than 3-fold change between the lowest and the highest growth rate was observed, the percentage of cells capable of invading Caco-2 cells varied more than 100-fold, that to form biofilms more than 30-fold, and the antibiotic MICs varied up to 512-fold, among the different strains. Results indicate that those strains with the highest cell adhesion ability were not always the most invasive ones and suggest that, in general terms, a higher stress resistance did not imply a reduced growth ability (rate). Similarly, no association between stress resistance and biofilm formation ability (except for acid stress) or antibiotic resistance (with minor exceptions) was found. Our data also suggest that, in Salmonella, acid stress resistance would be associated with virulence, since a positive correlation of that trait with adhesion and a negative correlation with invasion was found. This study contributes to a better understanding of the physiology of Salmonella and the relationship between bacterial stress resistance, growth ability, and virulence. It also provides new data regarding intra-specific variability of a series of phenotypic characteristics of Salmonella that are relevant from the food safety perspective.

Influence of the Initial Cell Number on the Growth Fitness of Salmonella Enteritidis in Raw and Pasteurized Liquid Whole Egg, Egg White, and Egg Yolk.

Salmonella growth in egg and egg products has been widely studied, but there are still some aspects that are not fully known. The objective of this work was to study the influence of the initial cell number on the growth fitness of Salmonella Enteritidis in raw and pasteurized egg products. Growth curves of five Salmonella Enteritidis strains in raw and pasteurized egg products, starting from different initial numbers, were obtained and fitted to the Baranyi and Roberts model. The results revealed that lower initial numbers led to longer lag phases (λ) and lower maximum specific growth rates (µmax) in raw liquid whole egg. Similar results were observed in raw egg white (except for one strain). Conversely, no influence (p > 0.05) of the initial concentration on Salmonella growth parameters in raw egg yolk was observed. On the other hand, no influence of the initial number of cells on Salmonella growth fitness in commercial pasteurized liquid whole egg was observed. The results obtained demonstrate that the disappearance of this initial-dose dependency phenomenon was dependent on the intensity of the thermal treatment applied. Finally, the influence of the initial number was, in general, lower in pasteurized than in raw egg white, but large differences among strains were observed.

Innovative Ultrasound-Assisted Approaches towards Reduction of Heavy Metals and Iodine in Macroalgal Biomass.

The aim of this work was to evaluate the potential of ultrasound (US), alone or in combination with mild heating and/or EDTA towards reduction of As, Cd, I, and Hg content of Laminaria hyperborea. Concentrations of As, Cd, I, and Hg of 56.29, 0.596, 7340, and <0.01 mg kg-1 of dry weight, respectively, were found in L. hyperborea blades. Treatment with US at 50 °C increased approx. 2-fold the amount of As released, although did not affect significantly the content of Cd or I, as compared to control (no US) samples. Reducing the temperature to 8 °C significantly decreased the effect of US, but heating at 80 °C did not cause a significant effect as compared to treatments at 50 °C. On the other hand, treatment with 0.1 N EDTA at 50 °C enhanced the percentage of Cd released by approximately 7-fold, regardless of sonication. In the present work, the combination of US and EDTA at 50 °C for 5 min led to a significant reduction of the As (32%), Cd (52%) and I (31%) content in L. hyperborea, thus improving the product's safety for consumers.

Impact of the Resistance Responses to Stress Conditions Encountered in Food and Food Processing Environments on the Virulence and Growth Fitness of Non-Typhoidal Salmonellae.

The success of Salmonella as a foodborne pathogen can probably be attributed to two major features: its remarkable genetic diversity and its extraordinary ability to adapt. Salmonella cells can survive in harsh environments, successfully compete for nutrients, and cause disease once inside the host. Furthermore, they are capable of rapidly reprogramming their metabolism, evolving in a short time from a stress-resistance mode to a growth or virulent mode, or even to express stress resistance and virulence factors at the same time if needed, thanks to a complex and fine-tuned regulatory network. It is nevertheless generally acknowledged that the development of stress resistance usually has a fitness cost for bacterial cells and that induction of stress resistance responses to certain agents can trigger changes in Salmonella virulence. In this review, we summarize and discuss current knowledge concerning the effects that the development of resistance responses to stress conditions encountered in food and food processing environments (including acid, osmotic and oxidative stress, starvation, modified atmospheres, detergents and disinfectants, chilling, heat, and non-thermal technologies) exerts on different aspects of the physiology of non-typhoidal Salmonellae, with special emphasis on virulence and growth fitness.

Direct Contact Ultrasound in Food Processing: Impact on Food Quality.

Consumers' demand for "minimally processed" products that maintain the "fresh-like" characteristics has increased in recent years. Ultrasound (US) is a non-thermal technology that enhances mass and energy transfer processes resulting in improved food quality. A new method of applying US to food without using a liquid or gaseous medium for the propagation of acoustic waves has recently been under research. It is known as direct contact US, since the food is directly placed on a plate where the transducers are located. In this type of systems, the main effect is not cavitation but acoustic vibration, which encourages mass and energy transfer processes due to the "sponge effect." Furthermore, as the product is not immersed in a liquid medium, the loss of hydrophilic nutritional compounds is reduced; systems such as these can thus be more easily implemented on an industrial level. Nevertheless, the very few studies that have been published about these systems mainly focus on dehydration and freezing. This article summarizes published research on the impact of direct contact US in nutritional and organoleptic quality of food in order to assess their potential to meet new market trends.

Variability in the heat resistance of Listeria monocytogenes under dynamic conditions can be more relevant than that evidenced by isothermal treatments.

Heterogeneity in the response of microbial cells to environmental conditions is inherent to every biological system and can be very relevant for food safety, potentially being as important as intrinsic and extrinsic factors. However, previous studies analyzing variability in the microbial response to thermal treatments were limited to data obtained under isothermal conditions, whereas in the reality, environmental conditions are dynamic. In this article we analyse both empirically and through mathematical modelling the variability in the microbial response to thermal treatments under isothermal and dynamic conditions. Heat resistance was studied for four strains of Listeria monocytogenes (Scott A, CECT 4031, CECT 4032 and 12MOB052), in three different matrices (buffered peptone water, pH 7 Mcllvaine buffer and semi-skimmed milk). Under isothermal conditions, between-strain and between-media variability had no impact in the heat resistance, whereas it was very relevant for dynamic conditions. Therefore, the differences observed under dynamic conditions can be attributed to the variability in the ability for developing stress acclimation. The highest acclimation was observed in strain CECT 4031 (10-fold increase of the D-value), while the lowest acclimation was observed in strain CECT 4032 (50% increase of the D-value). Concerning the different media, acclimation was higher in buffered peptone water and semi-skimmed milk than in Mcllvaine buffer of pH 7.0. To the knowledge of the authors, this is the first research work that specifically analyses the variability of microbial adaptation processes that take place under dynamic conditions. It highlights that microbial heat resistance under dynamic conditions are sometimes determined by mechanisms that cannot be observed when cells are treated in isothermal conditions (e.g. acclimation) and can also be affected by variability. Consequently, empirical evidence on variability gathered under isothermal conditions should be extrapolated with care for dynamic conditions.

Stress resistance of emerging poultry-associated Salmonella serovars.

In recent years, the on-farm prevalence of some poultry-related Salmonella serovars such as S. Kentucky, S. Heidelberg, S. Livingstone and S. Mbandaka has increased significantly, even replacing S. Enteritidis and S. Typhimurium as the most frequently isolated serovars in some production settings and countries. For this reason, the aim of this work was to determine the resistance to several stressing agents and food preservation technologies, in laboratory media and in egg products, of 4 strains of these emerging Salmonella serovars associated to poultry and poultry products and to make comparisons with 4 S. Enteritidis strains. First, the resistance to acid pH, hydrogen peroxide, NaCl, heat, HHP, PEF and UV of the 8 Salmonella strains studied was determined and compared in laboratory media. From this part of the study, it was concluded that variability in resistance to stress among the 8 studied strains varied depending on the investigated agent/technology. However, differences in resistance (2D-values) were always lower than 3.3-fold. Results obtained also indicated that the strains of the emerging serovars studied would display lower acid and NaCl resistance, higher heat resistance and similar oxidative, HHP, PEF and UV resistance than S. Enteritidis. Then, the resistance of these 8 strains was evaluated and compared in egg, egg products and poultry manure. For some agents -including osmotic stresses, UV and PEF- there was a very good correspondence between the results obtained in laboratory media and in real food matrices and poultry manure (r > 0.85; p < 0.01). A significant relationship was also found for acid and HHP resistance (p < 0.05) and a trend for heat resistance (p < 0.10). Therefore, in general terms, conclusions drawn from the study carried out in laboratory media - regarding intraspecific variability and the relative resistance of the different strains - might be extrapolated, although with caution, to real food scenarios. Results obtained in this investigation would help to better understand the physiology and ecology of Salmonella and to design better egg preservation strategies.

Understanding the occurrence of tailing in survival curves of Salmonella Typhimurium treated by pulsed electric fields.

This study aimed to gain more in-depth knowledge of the mechanisms involved in microbial inactivation by pulsed electric fields (PEF) to understand the tailing observed in survival curves of Salmonella Typhimurium (STCC 878). The comparison of the inactivation achieved by the application of one train of pulses with those obtained with pulses applied in two trains shows that the tail of the survival curves was a consequence of a transient increment of the microbial resistance to the effect of the electric field in a proportion of the cells. After some time following the application of the first pulse train, cells became again sensitive to the second train, and tailing tended to disappear. The required time was highly dependent on the characteristics of the incubation medium. Similar effects were observed when the treatments were validated on whole milk and orange juice. This study has demonstrated by the first time on microbial cells the benefits of splitting the delivered PEF treatment in two trains with a period of delay between them. Therefore, this insight opens up the possibility of developing new strategies to achieve the required inactivation levels to guarantee food safety by moderate PEF treatments.

Evolution of Polyphenolic Compounds and Sensory Properties of Wines Obtained from Grenache Grapes Treated by Pulsed Electric Fields during Aging in Bottles and in Oak Barrels.

The evolution of polyphenolic compounds and sensory properties of wines obtained from Grenache grapes, either untreated or treated with pulsed electric fields (PEF), in the course of bottle aging, as well as during oak aging followed by bottle aging, were compared. Immediately prior to aging in bottles or in barrels, enological parameters that depend on phenolic extraction during skin maceration were higher when grapes had been treated with PEF. In terms of color intensity, phenolic families, and individual phenols, the wine obtained with grapes treated by PEF followed an evolution similar to untreated control wine in the course of aging. Sensory analysis revealed that the application of a PEF treatment resulted in wines that are sensorially different: panelists preferred wines obtained from grapes treated with PEF. Physicochemical and sensory analyses showed that grapes treated with PEF are suitable for obtaining wines that require aging in bottles or in oak barrels.

Differences in resistance to different environmental stresses and non-thermal food preservation technologies among Salmonella enterica subsp. enterica strains.

In this work the resistance of 15 strains belonging to 11 serovars of Salmonella enterica subsp. enterica to several different environmental stresses (acid, hydrogen peroxide, NaCl and heat) and non-thermal food preservation technologies (HHP, PEF, UV) was determined and compared. Results obtained showed that differences in resistance among strains, quantified as 2D-values, varied less than 2.4-fold for all agents, including heat if S. senftenberg 775W is excluded from the analysis. These results also indicate that variability in resistance among strains of the same serovar was comparable to inter-serovar variability. Salmonella strains that were the most resistant to a given stress were not more resistant to other types of stress. Nevertheless, a positive correlation was observed between the resistance of Salmonella strains to oxidative and osmotic stress, as well as between UV and PEF resistance. These results would be especially helpful in defining safe food preservation processes and might be very useful for improving quantitative microbiological risk assessments of Salmonella in food products.

Potential of Pulsed Electric Fields for the preparation of Spanish dry-cured sausages.

The aim of this investigation was to lay the groundwork of the potential application of Pulsed Electric Fields (PEF) technology for accelerating the drying process of meat and meat products, and specifically in this work of Spanish dry-cured sausages "longaniza". PEF treatments were applied to pork loin samples, and the influence of different PEF parameters on the process were evaluated. An optimal PEF treatment of 1 kV/cm, 200 µs of pulse width and 28 kJ/kg was determined as the most suitable to electroporate meat cells and to improve water transfer by achieving a water content reduction of 60.4% in treated-meat samples dried at 4 °C. The influence of PEF on meat drying rate was also studied on minced pork and the results showed that with a particle size of 4.0 mm higher drying rates were achieved. To validate the results, Spanish cured sausages were prepared from treated and untreated minced pork and stuffed into gauzes and natural pork casings at pilot plant scale. After the curing process, the application of PEF to sausages stuffed into gauze reduced the drying time from 17 to 9-10 days, a reduction of 41-47%, confirming the effects described at lab scale and the potential of PEF for accelerating the sausage-drying process.

Dataset on the use of the Ratkowsky model for describing the influence of storage temperature on microbial growth in hake fillets (Merluccius merluccius) stored under MAP.

This article presents the results obtained after applying the Ratkowsky model for developing secondary models describing the influence of storage temperature on microbial growth in hake fillets packaged under a modified atmosphere (MAP) rich in CO2 (50% CO2/50% N2). For this purpose the growth parameters (λ, µmax) already calculated in the related article "Modelling microbial growth in Modified-Atmosphere-Packed hake (Merluccius merluccius) fillets stored at different temperatures" [1] were used. The data include the fit and goodness of the fit parameters calculated as well as the comparison between fitted and observed data.

Characterization of the Spoilage Microbiota of Hake Fillets Packaged Under a Modified Atmosphere (MAP) Rich in CO2 (50% CO2/50% N2) and Stored at Different Temperatures.

The aim of this study was to characterize the spoilage microbiota of hake fillets stored under modified atmospheres (MAP) (50% CO2/50% N2) at different temperatures using high-throughput 16S rRNA gene sequencing and to compare the results with those obtained using traditional microbiology techniques. The results obtained indicate that, as expected, higher storage temperatures lead to shorter shelf-lives (the time of sensory rejection by panelists). Thus, the shelf-life decreased from six days to two days for Batch A when the storage temperature increased from 1 to 7 °C, and from five to two days-when the same increase in storage temperature was compared-for Batch B. In all cases, the trimethylamine (TMA) levels measured at the time of sensory rejection of hake fillets exceeded the recommended threshold of 5 mg/100 g. Photobacterium and Psychrobacter were the most abundant genera at the time of spoilage in all but one of the samples analyzed: Thus, Photobacterium represented between 19% and 46%, and Psychrobacter between 27% and 38% of the total microbiota. They were followed by Moritella, Carnobacterium, Shewanella, and Vibrio, whose relative order varied depending on the sample/batch analyzed. These results highlight the relevance of Photobacterium as a spoiler of hake stored in atmospheres rich in CO2. Further research will be required to elucidate if other microorganisms, such as Psychrobacter, Moritella, or Carnobacterium, also contribute to spoilage of hake when stored under MAP.

Cellular events involved in E. coli cells inactivation by several agents for food preservation: A comparative study.

Traditional and novel technologies for food preservation are being investigated to obtain safer products and fulfil consumer demands for less processed foods. These technologies inactivate microorganisms present in foods through their action on different cellular targets, but the final cause of cell loss of viability often remains not well characterized. The main objective of this work was to study and compare cellular events that could play a role on E. coli inactivation upon exposure to treatments with technologies of different nature. E. coli cells were exposed to heat, high hydrostatic pressure (HHP), pulsed electric fields (PEF) and acid treatments, and the occurrence of several alterations, including presence of sublethal injury, membrane permeabilization, increased levels of reactive oxygen species (ROS), DNA damage and protein damage were studied. Results reflected differences among the relevance of the several cellular events depending on the agent applied. Sublethally injured cells appeared after all the treatments. Cells consistently recovered in a higher percentage in non-selective medium, particularly in minimal medium, as compared to selective medium; however this effect was less relevant in PEF-treated cells. Increased levels of ROS were detected inside cells after all the treatments, although their order of appearance and relationship with membrane permeabilization varied depending on the technology. A high degree of membrane permeabilization was observed in PEF treated cells, DNA damage appeared as an important target in acid treatment, and protein damage, in HHP treated cells. Results obtained help to understand the mode of action of food preservation technologies on bacterial cells.

Modelling microbial growth in modified-atmosphere-packed hake (Merluccius merluccius) fillets stored at different temperatures.

Market globalization and changes in purchasing habits pose a challenge to the fishery industry because of the short shelf life of fish products. In view of this scenario, it would be very helpful if tools capable of predicting the shelf-life of fish could be developed. Thus, the objective of this study was to employ a modelling approach capable of predicting the evolution of the microbiota of hake fillets packaged under a modified atmosphere (MAP) rich in CO2 (50% CO2 / 50% N2) when stored at temperatures ranging between 1 and 10 °C. Growth curves of ten microbial groups were obtained at four different temperatures and fitted with the Baranyi model. Photobacterium showed high growth rates in hake fillets (0.99 days-1 at 4 °C), similar to those of Shewanella, lactic acid bacteria, and non-specific microbial groups investigated, and significantly higher than those of Pseudomonas. Furthermore, no lag phase was observed for Photobacterium regardless of the temperature investigated. On the other hand, Enterobacteriaceae and moulds and yeasts displayed low growth fitness, and their counts increased by <1.5-2 Log10 cycles along the incubation period regardless of storage temperature. The influence of storage temperature on growth parameters (λ, µmax and Yend) was subsequently studied, and secondary models were developed for the eight most relevant microbial groups. All of the final equations developed in this study showed R2 values ≥0.90, and RMSE values ≤0.50. In addition, results obtained in this investigation strongly suggest that Photobacterium would be the main responsible microorganism for the spoilage of hake fillets stored under MAP conditions (50% CO2/50% N2) along the entire range of temperatures investigated (1-10 °C).

Protective effect of glutathione on Escherichia coli cells upon lethal heat stress.

Heat treatments are widely used by the food industry to obtain safe and stable products, therefore a deeper knowledge of its mode of action on microorganisms would allow a better profit of this technology. Heat shows a multitarget mechanism of action on bacteria, affecting various cellular structures, and causing unbalances in several homeostatic parameters. The aim of this work was to study the effect of glutathione on bacterial survival against heat treatments, in order to acquire knowledge about the mode of action of heat on bacterial cells. Cells were treated in presence or absence of added glutathione and the level of inactivation, soluble protein concentration, enzymatic activities, intracellular ROS level and membrane damages were studied. Results showed that glutathione protected microorganisms against heat inactivation. Moreover, glutathione in the treatment medium preserved intracellular enzyme activity, membrane structure and reduced ROS detection. Besides, glutathione decreased sublethal injury in E. coli. Data presented in this work add new knowledge about bacterial inactivation and survival by heat.