Elisa Gayán

Inactivation of Salmonella enterica by UV-C Light Alone and in Combination with Mild Temperatures

ABSTRACT 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.

Evaluation of pulsed electric fields technology for liquid whole egg pasteurization.

This investigation evaluated the lethal efficiency of pulsed electric fields (PEFs) to pasteurize liquid whole egg (LWE). To achieve this aim, we describe the inactivation of Salmonella Enteritidis and the heat resistant Salmonella Senftenberg 775 W in terms of treatment time and specific energy at electric field strengths ranging from 20 to 45 kV/cm. Based on our results, the target microorganism for this technology in LWE varied with intensity of the PEF treatment. For electric field strengths greater than 25 kV/cm, Salmonella Enteritidis was the most PEF-resistant strain. For this Salmonella serovar the level of inactivation depended only on the specific energy applied: i.e., 106, 272, and 472 kJ/kg for 1, 2, and 3 Log(10) reductions, respectively. The developed mathematical equations based on the Weibull distribution permit estimations of maximum inactivation level of 1.9 Log(10) cycles of the target Salmonella serovar in the best-case scenario: 250 kJ/kg and 25 kV/cm. This level of inactivation indicates that PEF technology by itself cannot guarantee the security of LWE based on USDA and European regulations. The occurrence of cell damage due to PEF in the Salmonella population opens the possibility of designing combined processes enabling increased microbial lethality in LWE.

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.

Design of a combined process for the inactivation of Salmonella Enteritidis in liquid whole egg at 55°C.

This paper is an evaluation of the lethal effectiveness of a successive application of pulsed electric fields (PEFs) and heat treatment in liquid whole egg (LWE) in the presence of different additives on the population of Salmonella serovar Enteritidis. Synergistic reductions of the Salmonella Enteritidis population were observed when LWE samples containing additives were treated with PEF (25 kV/cm; 100 and 200 kJ/kg), heat (55 °C), or PEF followed by heat. The presence of additives, such as 10 mM EDTA or 2% triethyl citrate, increased the PEF lethality 1 log₁₀ cycle and generated around 1.5 log₁₀ cycles of cell damage, resulting in the reduction of undamaged cells of 4.4 and 3.1 log₁₀ cycles, respectively. The application of PEF followed by heat treatment significantly (p < 0.05) reduced D(55 ºC) from 3.9 ± 0.2 min in LWE to 1.40 ± 0.06 min or 0.24 ± 0.02 min in the presence of 10 mM EDTA or 2% triethyl citrate, respectively. A PEF treatment of 25 kV/cm and 200 kJ/kg followed by a heat treatment of 55 °C and 2 min reduced more than 8 log₁₀ cycles of the population of Salmonella Enteritidis in LWE combined with 2% triethyl citrate, with a minimal impact on its protein soluble content. The heat sensitizing effect of PEF treatments in the presence of 2% triethyl citrate on the Salmonella population could enable LWE producers to reduce the temperature or processing time of thermal treatments (current standards are 60 °C for 3.5 min in the United States), increasing the level of Salmonella inactivation and retaining the quality of non-treated LWE.

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.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.

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 mediums 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.

Hurdle approach to increase the microbial inactivation by high pressure processing: Effect of essential oils

Consumer demand for improved quality and fresh-like food products has led to the development of new nonthermal preservation methods. High pressure processing (HPP) is currently the novel nonthermal technology best established in the food processing industry. However, many potential HPP applications would require long treatment times to ensure an adequate inactivation level of pathogens and spoilage microorganisms. High hydrostatic pressure and the addition of essential oils (EOs) have similar effects on microbial structures and thus they may act synergistically on the inactivation of microorganisms. Therefore, the combination of high hydrostatic pressure with EOs is a promising alternative to expand the HPP food industry. In this work, findings on this scarcely investigated hurdle option have been reviewed with a focus on the mechanisms involved. The main mechanisms involved are as follows: (1) membrane permeability induced by HPP and EOs facilitating the uptake of EOs by bacterial cells; (2) generation of reactive oxygen species via the Fenton reaction; (3) impairment of the proton motive force and electron flow; and (4) disruption of the protein–lipid interaction at the cell membrane altering numerous cellular functions. The effectiveness of a specific EO in enhancing the microbial inactivation level achieved by HPP treatments depends on the microbial ecology of the food product, the molecular mechanisms of the microbial inactivation by HPP, and the mode of action of the EO being used.

Effect of Pressure-Induced Changes in the Ionization Equilibria of Buffers on Inactivation of Escherichia coli and Staphylococcus aureus by High Hydrostatic Pressure

ABSTRACT Survival rates of Escherichia coli and Staphylococcus aureus after high-pressure treatment in buffers that had large or small reaction volumes (ΔV°), and which therefore underwent large or small changes in pH under pressure, were compared. At a low buffer concentration of 0.005 M, survival was, as expected, better in MOPS (morpholinepropanesulfonic acid), HEPES, and Tris, whose ΔV° values are approximately 5.0 to 7.0 cm3 mol−1, than in phosphate or dimethyl glutarate (DMG), whose ΔV° values are about −25 cm3 mol−1. However, at a concentration of 0.1 M, survival was unexpectedly better in phosphate and DMG than in MOPS, HEPES, or Tris. This was because the baroprotective effect of phosphate and DMG increased much more rapidly with increasing concentration than it did with MOPS, HEPES, or Tris. Further comparisons of survival in solutions of salts expected to cause large electrostriction effects (Na2SO4 and CaCl2) and those causing lower electrostriction (NaCl and KCl) were made. The salts with divalent ions were protective at much lower concentrations than salts with monovalent ions. Buffers and salts both protected against transient membrane disruption in E. coli, but the molar concentrations necessary for membrane protection were much lower for phosphate and Na2SO4 than for HEPES and NaCl. Possible protective mechanisms discussed include effects of electrolytes on water compressibility and kosmotropic and specific ion effects. The results of this systematic study will be of considerable practical significance in studies of pressure inactivation of microbes under defined conditions but also raise important fundamental questions regarding the mechanisms of baroprotection by ionic solutes.

Continuous-Flow UV Liquid Food Pasteurization: Engineering Aspects

Ultraviolet (UV)-C treatments are a promising technology for liquid food pasteurization as an alternative to heat treatments. However, the design of efficient UV reactors to reduce pertinent microorganisms and comply with current food safety goals is still an engineering challenge due to the low penetration depth of UV light in liquid foods with high UV absorbance and suspended particles, and the variations in the residence time of the product in the UV reactors. This review focuses on physical aspects of UV radiation related to the essential product and processing parameters for the design of UV reactors. The UV equipment available for liquid food processing is described and the main drawbacks and advantages are discussed.

NON-THERMAL PROCESSING | Pulsed UV Light

Pulsed ultraviolet (PUV) light involves the use of high-peak-power pulsed light of short duration and a broad spectrum ranging from ultraviolet to infrared wavelengths. This technology has been reported to have the potential to inactivate a broad range of spoilage and pathogenic microorganisms by limiting the negative effects on product quality and nutritional value. This chapter provides a comprehensive overview of the state of the art of microbial inactivation using PUV by considering its advantages and limitations.