Geobacillus stearothermophilus STCC4517 spore suspensions showed survival curves with shoulder phenomena independent of sporulation temperature and pH, whose duration was an exponential function of treatment temperature.

Deviations in linearity in survival curves are common in inactivation kinetics during heat treatment. These might lead one to underestimate how effective thermal treatment is. In previous research we reported a relationship between decimal reduction time values (DT) and shoulder lengths (Sl) of survival curves which was characteristic of each microorganism. However, the impact of other factors such as sporulation temperature and pH of the treatment media on shoulder length is still not known. The objective of this research was to evaluate the effect of sporulation temperature (45, 55 and 65 °C) and pH (4.0, 5.0, 6.0 and 7.0) treatment has on the profile of survival curves and on the relationship between Sl/DT of G. stearothermophilus STCC 4517. The results obtained demonstrated that all the spore suspensions, independently of sporulation temperature and pH, showed survival curves with shoulder phenomena, whose duration was an exponential function of treatment temperature. Although both parameters had a significant effect on heat resistance, the relationship between the shoulder length and DT values was constant at all pHs for spores produced at the optimum sporulation temperature.

Impact of shoulders on the calculus of heat sterilization treatments with different bacterial spores.

To date, heat is still the most used technology in food preservation. The calculus of heat treatments is usually based on Bigelow observations i.e. treatment time is an exponential function of the heat treatment temperature. However, a number of researchers have reported deviations from linearity in heat inactivation curves that caused errors in the calculus. This research was designed to evaluate the variability of shoulder length among different sporulated species, the impact of treatment temperature on these shoulders and the relationship between the traditional DT value and shoulder length. The heat inactivation kinetics of five bacterial spores of importance for the food industry was evaluated. B. weihenstephanensis and B. cereus did not show shoulders and DT values calculated ranged from 0.99 to 0.23 and from 1.33 to 0.56 respectively at temperatures from 100 to 102.5 °C. On the other side B. subtilis, B. licheniformis and G. stearothermophilus showed shoulders of 1.75-0.42, 1.92-0.43 and 3.22-0.78 and DT values of 1.52-0.32, 2.12-0.59 and 2.22-0.48 respectively in the range of temperatures tested. From the results obtained it was concluded that the presence and magnitude of shoulders depended on the bacterial spore species, the longest being those on the bacterial spores which showed greatest heat resistance. It has also been proved that shoulder lengths vary with treatment temperature in the same proportion of traditional DT values, with the relationship Sl/DT being constant. Thus, an equation which included the constant Sl/DT was proposed.

Heat resistance, membrane fluidity and sublethal damage in Staphylococcus aureus cells grown at different temperatures.

In this work the influence of growth temperature (10-42 °C) on Staphylococcus aureus heat resistance was studied, and its relationship with the ability of cells to repair sublethal damages and with membrane fluidity was evaluated. Non-linear, convex from above survival curves were obtained, and therefore a special case of the Baranyi model was used to fit them. For exponential phase cells, heat resistance did not change with growth temperature in the range between 10 and 37 °C, but cells grown at 42 °C were significantly more resistant, showing D58 and shoulder length (sl58) values 2.5 and 4 times greater than the others, respectively. For stationary growth phase cells, an increase in growth temperature above 20 °C resulted in an increase in D58 values, and cells grown at 42 °C also displayed the highest D58 and sl58 values. The increased heat resistance at 58 °C of stationary growth phase cells grown at higher temperatures was coincident with the appearance of a higher proportion of sublethally damaged cells capable of recovery and outgrowth in non-selective medium. Membrane fluidity was measured at treatment temperatures, and it was observed that those cells with more rigid membranes displayed greater heat resistance (Pearson coefficient = 0.969***). Additionally, S. aureus cells whose membrane was fluidized through exposure to benzyl alcohol were notably sensitized against the action of heat, in a concentration-dependent manner. Results obtained in this research indicate that membrane physical state could be an important factor determining the survival capacity of bacterial cells to a heat treatment.

Crab-meat-isolated psychrophilic spore forming bacteria inactivation by electron beam ionizing radiation.

The present work was performed to evaluate the potential of electron beam ionizing radiation for the inactivation of three psychrophilic spore forming bacteria (Bacillus mycoides, Bacillus weihenstephanensis and Psychrobacillus psychrodurans) isolated from ready-to-eat brown crab (Cancer pagurus). Inactivation curves for the three spores were performed in both types of crab meat, brown and white. Also the effect of pH and water activity (aw) on the lethal efficacy of ionizing radiation, for the three different psychrophilic spore forming bacteria, was evaluated. The effects of pH, aw and their possible interactions were assessed in citrate-phosphate buffers of different pH, ranging between 7 and 4, and aw, ranging from <0.99 to 0.80. A reduction of aw increased the spores resistance between >0.99 and 0.90, while an aw reduction from 0.90 to 0.80 had a minor impact on their resistance. In contrast to aw, the effect of pH showed a greater variability depending on the spore species. While pH did not affect the resistance of B. weihenstephanensis at any aw, B. mycoides showed slightly higher resistance at pH 5.5 at aw of 0.90 and 0.80. pH showed a significant effect on the resistance of P. psychrodurans. For the two types of crab meat, slightly differences were observed in 6D values. B. weihenstephanensis was the most resistant, requiring 7.3-7.6 kGy to inactivate 6 Log10-cycles of this spore forming bacterium, while for B. mycoides and P. psychrodurans 6.1-6.3 and 5.4-5.3 kGy respectively were necessary to reach the same inactivation level in crab meat. An agreement between spore resistance in crab meats and lab media, with similar characteristics in pH and aw, was also observed. The results obtained in this research demonstrated the potential for ionizing radiation to achieve an appropriate inactivation level of spores naturally present in brown crab with the application of doses lower than 10 kGy.

Evaluation of the potential of ultrasound technology combined with mild temperatures to reduce cadmium content of edible crab (Cancer pagurus).

The consumption of crustaceans is correlated with certain health risks, particularly due to several highly toxic elements they contain, including cadmium (Cd). Although Cd content in one sole crab generally exceeds the total weekly recommended intake of cadmium as established by EFSA (especially in brown meat), efficient modern strategies to reduce Cd content in crabs still have not yet been developed. The objective of this research was therefore to evaluate the potential use of ultrasound technology in combination with temperature (50°-80 °C) with the purpose of releasing Cd from brown crab (Cancer pagurus), thereby reducing the Cd content in its meat. Female crabs were immersed in a water bath at 50, 65, and 80 °C in presence or absence of ultrasound; Cd concentration in the water was monitored along time. At the end of the process, Cd content in brown and white crab meat was likewise quantified. Treatment temperature did not bear an influence on the release of Cd in absence of ultrasound, but proved to be an important variable when ultrasound assisted the process. Ultrasound increased Cd release rates 8.7-, 2.1- and 2.7-fold in conjunction with the treatments at 50, 65 and 80 °C, respectively. The maximum percentage of Cd extracted (22.8%) was observed at 50 °C for an ultrasound input power of 200 W. These results have demonstrated for the first time that the application of ultrasound during the crab-cooking process could serve as an effective physical procedure for reducing the Cd content of crabs, thereby improving the product's safety for consumers.

Synergistic effect of ultrasonic waves under pressure at mild temperatures (MTS) in yeast inactivation.

Ultrasonic treatments are one of the new technologies for microbial inactivation that could serve as an alternative for food preservation. However, decimal reduction times for most of microbial species generally exceed 1 min. Therefore, combined processes have been designed, based on the simultaneous application of ultrasonic waves under pressure at moderate temperatures (mano-thermo-sonication process, MTS). The aim of this study was mathematically quantify the synergism of MTS treatments on S. bayanus and different microbial groups including vegetative cells and bacterial spores and compare them. Results show that the lethal effect of MTS treatments may have both, additive (A. hydrophila, Y. enterocolitica) or synergistic effect (S. bayanus, L. monocytogenes, Salmonella spp., Bacillus spp.). The synergistic effect increases with temperature until reaching a maximum and then descending. A big synergistic effect was observed in yeasts and bacterial spores while lower synergy was observed in bacterial vegetative cells. The more heat resistant bacterial species showed higher synergistic effect of the MTS treatments.

Bivalent IAP antagonists, but not monovalent IAP antagonists, inhibit TNF-mediated NF-κB signaling by degrading TRAF2-associated cIAP1 in cancer cells.

The inhibitor of apoptosis (IAP) proteins have pivotal roles in cell proliferation and differentiation, and antagonizing IAPs in certain cancer cell lines results in induction of cell death. A variety of IAP antagonist compounds targeting the baculovirus IAP protein repeat 3 (BIR3) domain of cIAP1have advanced into clinical trials. Here we sought to compare and contrast the biochemical activities of selected monovalent and bivalent IAP antagonists with the intent of identifying functional differences between these two classes of IAP antagonist drug candidates. The anti-cellular IAP1 (cIAP1) and pro-apoptotic activities of monovalent IAP antagonists were increased by using a single covalent bond to combine the monovalent moieties at the P4 position. In addition, regardless of drug concentration, treatment with monovalent compounds resulted in consistently higher levels of residual cIAP1 compared with that seen following bivalent compound treatment. We found that the remaining residual cIAP1 following monovalent compound treatment was predominantly tumor necrosis factor (TNF) receptor-associated factor 2 (TRAF2)-associated cIAP1. As a consequence, bivalent compounds were more effective at inhibiting TNF-induced activation of p65/NF-κB compared with monovalent compounds. Moreover, extension of the linker chain at the P4 position of bivalent compounds resulted in a decreased ability to degrade TRAF2-associated cIAP1 in a manner similar to monovalent compounds. This result implied that specific bivalent IAP antagonists but not monovalent compounds were capable of inducing formation of a cIAP1 E3 ubiquitin ligase complex with the capacity to effectively degrade TRAF2-associated cIAP1. These results further suggested that only certain bivalent IAP antagonists are preferred for the targeting of TNF-dependent signaling for the treatment of cancer or infectious diseases.

Application of ultrasound in combination with heat and pressure for the inactivation of spore forming bacteria isolated from edible crab (Cancer pagurus).

This research was performed to characterize the resistance of three different bacterial spore species isolated from pasteurized edible crab (Cancer pagurus) meat to heat treatments and to assess the potential of manosonication (MS) and manothermosonication (MTS) as an alternative for their inactivation. The spore-forming bacteria used in this study were Bacillus mycoides, Bacillus weihenstephanensis and Psychrobacillus psychrodurans. The thermal resistance of these three species was determined at different temperatures ranging from 80 to 110 °C and their resistance to ultrasound under pressure from 35 to 95 °C. Ginafit Excel tool was used to fit the Geeraerd's 'Log-linear + shoulder' and Bigelow & Easty's equations to the survival curves for heat and MS/MTS treatments. From the results obtained it can be concluded that the profile of the survival curves either for heat or for ultrasound treatments depended on the bacterial spore species. When shoulders were detected in the inactivation curves for heat they were also present in the curves for MS/MTS treatments, although the application of an ultrasonic field reduced the shoulder length. B. weihenstephanensis was found to be the most resistant species to heat, requiring 1.4 min to reduce 4log10 cycles at 107.5 °C (zT=7.1 °C) while B. mycoides was the most sensitive requiring 1.6 min at 95 °C (zT=9.1 °C). By contrast, B. mycoides was the most resistant to MS. The efficiency of the combination of ultrasonic waves under pressure with heat (MTS) for bacterial spore inactivation was directly correlated with the thermal resistance. Indeed, MTS showed a synergistic effect for the inactivation of the three spores. The highest percentage of synergism corresponded to the spore species with higher zT value (B. mycoides), but the highest temperature at which this synergism was detected corresponded to the most heat tolerant spore species (B. weihenstephanensis). This study revealed that MTS treatment is capable of inactivating spore-forming bacteria and that the inactivation efficiency of the combined treatment is correlated with the thermal resistance of the spore species.

UV-Heat Treatments for the Control of Foodborne Microbial Pathogens in Chicken Broth.

This investigation established the process criteria for using UV-C light and mild heat (UV-H treatment) to inactivate 5-Log10 cycles (performance criterion) of common foodborne pathogen populations, Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus, when inoculated in chicken broth. To define the target microorganism and the proper UV-H treatment conditions (including UV dose, treatment time, and temperature) that would achieve the stated performance criterion, mathematical equations based on Geeraerd's model were developed for each microorganism. For the sake of comparison, inactivation equations for heat treatments were also performed on the same chicken broth and for the same microorganisms. L. monocytogenes was the most UV-H resistant microorganism at all temperatures, requiring a UV dose between 6.10 J/mL (5.6 min) and 2.26 J/mL (2.09 min) to achieve 5-Log10 reductions. In comparison with UV treatments at room temperatures, the combination of UV and mild heat allowed both the UV dose and treatment time to be reduced by 30% and 63% at 55 °C and 60 °C, respectively. Compared to heat treatments, the UV-H process reduced the heating time for 5-Log10 reductions of all the investigated microorganisms in chicken broth from 20-fold to 2-fold when the operating temperature varied from 53 to 60 °C.

Osmotolerance provided by the alternative sigma factors σB and rpoS to Staphylococcus aureus and Escherichia coli is solute dependent and does not result in an increased growth fitness in NaCl containing media.

The aim of this work was to examine the role of the alternative general stress sigma factors σ(B) and rpoS on the ability of Staphylococcus aureus and Escherichia coli, respectively, to grow in liquid and solid media of different osmolarity. For this purpose, S. aureus strain Newman and its isogenic ΔsigB mutant IK84 and E. coli strain BJ4 and its isogenic ΔrpoS mutant BJ4L1 were grown in media (TSBYE) with different concentrations of NaCl. Growth parameters (lag phase duration, growth rate and maximum number of microorganisms) and limiting growth concentrations (Maximum Non-Inhibitory Concentration - MNIC - and Minimum Inhibitory Concentration - MIC-) were determined. The mechanisms underlying the differences observed between parental and mutant strains were also explored. The absence of the sigma factors σ(B) and rpoS led to a decrease in the MNICs and MICs calculated for S. aureus and E. coli, respectively. Conversely, neither σ(B) nor rpoS provided with increased growth fitness to S. aureus and E. coli cells at NaCl concentrations up to 1.36M and 1M, respectively. The decreased osmotolerance of the σ(B) and rpoS deficient strains, as compared to their parental strains, was compensated by the addition of glycine-betaine (1mM) to the growth medium. It was also observed that the decreased tolerance to NaCl of the mutant strains was coincident with a decreased tolerance to sucrose, KCl, and LiCl but not to glycerol, MgCl2, and CaCl2. Results obtained also demonstrate that the increased osmotolerance of stationary growth phase E. coli cells, as compared to exponential growth phase ones, would be due to the activation of both rpoS-independent and rpoS-dependent mechanisms. This work will help to understand the mechanisms of bacterial resistance to osmotic stress and the role of the alternative sigma factors σ(B) and rpoS in this process.

Environmental and biological factors influencing the UV-C resistance of Listeria monocytogenes.

In this investigation, the effect of microbiological factors (strain, growth phase, exposition to sublethal stresses, and photorepair ability), treatment medium characteristics (pH, water activity, and absorption coefficient), and processing parameters (dose and temperature) on the UV resistance of Listeria monocytogenes was studied. The dose to inactivate 99.99% of the initial population of the five strains tested ranged from 21.84 J/mL (STCC 5672) to 14.66 J/mL (STCC 4031). The UV inactivation of the most resistant strain did not change in different growth phases and after exposure to sublethal heat, acid, basic, and oxidative shocks. The pH and water activity of the treatment medium did not affect the UV resistance of L. monocytogenes, whereas the inactivation rate decreased exponentially with the absorption coefficient. The lethal effect of UV radiation increased synergistically with temperature between 50 and 60 °C (UV-H treatment). A UV-H treatment of 27.10 J/mL at 55 °C reached 2.99 and 3.69 Log10 inactivation cycles of L. monocytogenes in orange juice and vegetable broth, and more than 5 Log10 cycles in apple juice and chicken broth. This synergistic effect opens the possibility to design UV combined processes for the pasteurization of liquid foods with high absorptivity.

Bacterial maximum non-inhibitory and minimum inhibitory concentrations of different water activity depressing solutes.

The NaCl MNICs (maximum non-inhibitory concentrations) and MICs (minimum inhibitory concentrations) for growth of various strains of six bacterial species were determined and then compared with those obtained for seven other solutes. The influence of prior growth conditions on the MNICs and MICs was also evaluated. No significant changes on the MNICs and MICs were found among the strains studied within each species. Among all factors investigated, only growth phase -for Gram-negatives- and growth at high NaCl concentrations led to a change in the NaCl MNICs. Species could be classified depending on its NaCl MNICs and MICs (in decreasing order) as follows: Staphylococcus aureus, Listeria monocytogenes, Cronobacter sakazakii, Enterococcus faecium, Escherichia coli and Salmonella Typhimurium. Similar results were obtained for KCl, LiCl, and sodium acetate, but not for the remaining solutes investigated (sucrose, glycerol, MgCl2 and CaCl2). Results obtained indicate that, in general, Gram-negatives showed lower MNICs and MICs than Gram-positives for all the solutes, S. aureus being the most solute tolerant microorganism. When compared on a molar basis, glycerol showed the highest MNICs and MICs for all the microorganisms -except for S. aureus- and LiCl the lowest ones. NaCl MNICs and MICs were not significantly different from those of KCl when compared on a molar basis. Therefore, the inhibitory action of NaCl could not be linked to the specific action of Na(+). Results also showed that the Na(+) tolerance of some species was Cl(-) dependent whereas for others it was not, and that factors others than aw-decrease contribute to the inhibitory action of LiCl, CaCl2 and MgCl2.

Microbiological aspects related to the feasibility of PEF technology for food pasteurization.

Processing unit operations that seek to inactivate harmful microorganisms are of primary importance in ascertaining the safety of food. The capability of pulsed electric fields (PEF) to inactivate vegetative cells of microorganisms at temperatures below those used in thermal processing makes this technology very attractive as a nonthermal pasteurization process for the food industry. Commercial exploitation of this technology for food pasteurization requires the identification of the most PEF-resistant microorganisms that are of concern to public health. Then, the treatment conditions applicable at industrial scale that would reduce the population of these microorganisms to a level that guarantees food safety must be defined. The objective of this paper is to critically compile recent, relevant knowledge with the purpose of enhancing the feasibility of using PEF technology for food pasteurization and underlining the required research for designing PEF pasteurization processes.

Selection of process conditions by risk assessment for apple juice pasteurization by UV-heat treatments at moderate temperatures.

The effect of bactericidal UV-C treatments (254 nm) on Escherichia coli O157:H7 suspended in apple juice increased synergistically with temperature up to a threshold value. The optimum UV-C treatment temperature was 55 °C, yielding a 58.9% synergistic lethal effect. Under these treatment conditions, the UV-heat (UV-H55 °C) lethal variability achieving 5-log reductions had a logistic distribution (α = 37.92, ß = 1.10). Using this distribution, UV-H55 °C doses to achieve the required juice safety goal with 95, 99, and 99.9% confidence were 41.17, 42.97, and 46.00 J/ml, respectively, i.e., doses higher than the 37.58 J/ml estimated by a deterministic procedure. The public health impact of these results is that the larger UV-H55 °C dose required for achieving 5-log reductions with 95, 99, and 99.9% confidence would reduce the probability of hemolytic uremic syndrome in children by 76.3, 88.6, and 96.9%, respectively. This study illustrates the importance of including the effect of data variability when selecting operational parameters for novel and conventional preservation processes to achieve high food safety standards with the desired confidence level.

Resistance of Staphylococcus aureus to UV-C light and combined UV-heat treatments at mild temperatures.

In this investigation, the resistance of enterotoxigenic Staphylococcus aureus to short-wave ultraviolet light (UV-C) and to combined UV C-heat (UV-H) treatments in buffers and in liquid foods with different physicochemical characteristics was studied. Microbial resistance to UV-C varied slightly among the S. aureus strains tested. The UV-C resistance of S. aureus increased in the entry of stationary growth phase, which in part was due to the expression of the alternative sigma factor σ(B). The UV-C resistance of S. aureus was independent of the treatment medium's pH and water activity, but it decreased exponentially as the absorption coefficient increased. UV-C bactericidal efficacy in liquids of high absorption coefficients was improved synergistically when combined with a mild heat treatment at temperatures ranging from 50.0 to 57.5 °C. pH of the treatment medium modified the lethality of UV-H treatments and therefore the temperature of maximum synergy. The advantage of combined UV-H treatments was demonstrated in fruit juices and vegetable and chicken broths, inactivating 5 Log10 cycles of S. aureus by applying UV-C treatments of 27.1 mJ/L for 3.6 min at 52.5 °C or 13.6 mJ/L for 1.8 min at 55.0 °C.

Emergence of pulsed electric fields resistance in Salmonella enterica serovar Typhimurium SL1344.

In this investigation we selected and isolated a culture derived from Salmonella enterica serovar Typhimurium SL1344 with stable increased resistance to pulsed electric fields (PEF) after repeated rounds of PEF treatment and outgrowth of survivors. The resulting culture showed a higher resistance to PEF treatments under different treatment conditions. The acquisition of PEF resistance was only observed in stationary phase cells. The cytoplasmic membrane of the resistant variant showed a higher resilience against PEF treatments, since a lower permeabilization degree was observed after PEF treatments, in comparison to the parental strain. Resistance to PEF was also accompanied by a higher tolerance to acidic pH, hydrogen peroxide and ethanol, but not to heat. The occurrence of a PEF resistant variant in S. enterica serovar Typhimurium SL1344 emphasizes the need to further study the mechanisms of inactivation and resistance by PEF for an adequate design of safe treatments.

Mechanism of the synergistic inactivation of Escherichia coli by UV-C light at mild temperatures.

UV light only penetrates liquid food surfaces to a very short depth, thereby limiting its industrial application in food pasteurization. One promising alternative is the combination of UV light with mild heat (UV-H), which has been demonstrated to produce a synergistic bactericidal effect. The aim of this article is to elucidate the mechanism of synergistic cellular inactivation resulting from the simultaneous application of UV light and heat. The lethality of UV-H treatments remained constant below ∼45°C, while lethality increased exponentially as the temperature increased. The percentage of synergism reached a maximum (40.3%) at 55°C. Neither the flow regimen nor changes in the dose delivered by UV lamps contributed to the observed synergism. UV-H inactivation curves of the parental Escherichia coli strain obtained in a caffeic acid selective recovery medium followed a similar profile to those obtained with uvrA mutant cells in a nonselective medium. Thermal fluidification of membranes and synergistic lethal effects started around 40 to 45°C. Chemical membrane fluidification with benzyl alcohol decreased the UV resistance of the parental strain but not that of the uvrA mutant. These results suggest that the synergistic lethal effect of UV-H treatments is due to the inhibition of DNA excision repair resulting from the membrane fluidification caused by simultaneous heating.

Inactivation of Salmonella enterica by UV-C light alone and in combination with mild temperatures.

The aim of this investigation was to study the efficacy of the combined processes of UV light and mild temperatures for the inactivation of Salmonella enterica subsp. enterica and to explore the mechanism of inactivation. The doses to inactivate the 99.99% (4D) of the initial population ranged from 18.03 (Salmonella enterica serovar Typhimurium STCC 878) to 12.75 J ml(-1) (Salmonella enterica serovar Enteritidis ATCC 13076). The pH and water activity of the treatment medium did not change the UV tolerance, but it decreased exponentially by increasing the absorption coefficient. An inactivating synergistic effect was observed by applying simultaneous UV light and heat treatment (UV-H). A less synergistic effect was observed by applying UV light first and heat subsequently. UV did not damage cell envelopes, but the number of injured cells was higher after a UV-H treatment than after heating. The synergistic effect observed by combining simultaneous UV and heat treatment opens the possibility to design combined treatments for pasteurization of liquid food with high UV absorptivity, such as fruit juices.

Inactivation of Salmonella spp. in liquid whole egg using pulsed electric fields, heat, and additives.

This paper evaluates the lethal effectiveness on 7 different Salmonella serovars of the application, in static and continuous conditions, of pulsed electric fields (PEF) followed by heat treatments in liquid whole egg (LWE) with additives (EDTA or triethyl citrate-TC-). Compared to heat treatments, the PEF (25 kV/cm and 75-100 kJ/kg) followed by heat (52°C/3.5', 55°C/2', or 60°C/1') in LWE with 2% TC permitted the reduction of heat treatment time from 92 fold at 52°C to 3.4 fold at 60°C, and 4.8 fold at 52°C in LWE with EDTA for a 9-Log(10) reduction of the population of Salmonella Enteritidis. The new designed treatments inactivated more than 5 Log(10) cycles of Salmonella serovars Dublin, Enteritidis 4300, Enteritidis 4396, Typhimurium, Typhi, Senftenberg, and Virchow, both in static and continuous conditions. Conversely, current heat pasteurization treatments of 60°C/3.5' and 64°C/2.5' reduced 5 Log(10) cycles of various serovars of Salmonella but only 2 and 3-4 Log(10) cycles of Salmonella Senftenberg and Salmonella Enteritidis 4396, respectively. Soluble protein content (SPC) decreased 1.8%, 1.3%, and 5.0% after the successive application of PEF followed by heat at 52, 55, and 60°C in the presence of 2% TC, respectively, whereas 1.6% and 9.4% of SPC were reduced after heat pasteurization at 60 and 64°C, respectively. Results indicate that designed treatments could be an alternative to current heat pasteurization of LWE as showed higher lethal effectiveness against Salmonella serovars with a similar or even lower decrement of the soluble protein content.

Development of resistance in Cronobacter sakazakii ATCC 29544 to thermal and nonthermal processes after exposure to stressing environmental conditions.

AIMS: The objective was to study the response of Cronobacter sakazakii ATCC 29544 cells to heat, pulsed electric fields (PEF), ultrasound under pressure (Manosonication, MS) and ultraviolet light (UV-C) treatments after exposure to different sublethal stresses that may be encountered in food-processing environments. METHODS AND RESULTS: Cronobacter sakazakii stationary growth-phase cells (30°C, 24h) were exposed to acid (pH 4·5, 1h), alkaline (pH 9·0, 1h), osmotic (5% NaCl, 1h), oxidative (0·5mmoll(-1) H(2) O(2), 1h), heat (47·5°C, 1h) and cold (4°C, 4h) stress conditions and subjected to the subsequent challenges: heat (60°C), PEF (25kVcm(-1) , 35°C), MS (117µm, 200kPa, 35°C) and UV-C light (88·55mWcm(-2), 25°C) treatments. The inactivation kinetics of C. sakazakii by the different technologies did not change after exposure to any of the stresses. The combinations of sublethal stress and lethal treatment that were protective were: heat shock-heat, heat shock-PEF and acid pH-PEF. Conversely, the alkaline shock sensitized the cells to heat and UV-C treatments, the osmotic shock to heat treatments and the oxidative shock to UV-C treatments. The maximum adaptive response was observed when heat-shocked cells were subjected to a heat treatment, increasing the time to inactivate 99·9% of the population by 1·6 times. CONCLUSIONS: Cronobacter sakazakii resistance to thermal and nonthermal preservation technologies can increase or decrease as a consequence of previous exposure to stressing conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The results help in understanding the physiology of the resistance of this emerging pathogen to traditional and novel preservation technologies.