Inactivation of Escherichia coli O157: H7 in foods by emerging technologies: a review.

Escherichia coli O157: H7 is a representative foodborne pathogen that causes haemorrhagic colitis, bloody diarrhea, and fatal haemolytic uraemic syndrome. Previously, only conventional heat treatment was used to pasteurised food; however, this method decreases food quality, including colour change, denatures proteins, and causes lipid oxidation. Therefore, emerging technologies to inactivate pathogens in food that affect food quality minimally have been researched and developed. This review aims to compile research since 2018 and briefly describe the inactivation mechanisms of emerging technologies such as microwave, radio frequency, ohmic heating, superheated steam, ionising radiation (gamma irradiation, electron beam, and X-rays), high pressure, ultraviolet light, pulsed light, ultrasound, gas treatment, plasma, and combination treatments. Pulsed electric field and electrolysed water were excluded because few research papers were published after 2018. In addition, the shortcomings of emerging technologies in the control of E. coli O157: H7 and the directions for emerging technology research are presented. Taking advantage of emerging technologies with many benefits will significantly improve food safety.

Antibacterial Activity of Caffeic Acid Combined with UV-A Light against Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes.

The aim of this study was to evaluate the antibacterial activity of caffeic acid (CA), which is a natural polyphenol, combined with UV-A light against the representative foodborne bacteria Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes. Data regarding the inactivation of these bacteria and its dependence on CA concentration, light wavelength, and light dose were obtained. E. coli O157:H7 and Salmonella Typhimurium were reduced to the detection limit when treated with 3 mM CA and UV-A for 3 J/cm2 and 4 J/cm2, respectively, and 5 J/cm2 treatment induced 3.10 log reduction in L. monocytogenes. To investigate the mechanism for inactivation of Salmonella Typhimurium and L. monocytogenes, measurement of polyphenol uptake, membrane damage assessment, enzymatic activity assay, and transmission electron microscopy (TEM) were conducted. It was revealed that CA was significantly (P < 0.05) absorbed by bacterial cells, and UV-A light allowed a higher uptake of CA for both pathogens. Additionally, CA plus UV-A treatment induced significant (P < 0.05) cell membrane damage. In the enzymatic activity assay, the activities of both pathogens were reduced by CA, and a greater reduction occurred by use of CA plus UV-A. Moreover, transmission electron microscopy (TEM) images indicated that CA plus UV-A treatment notably destroyed the intercellular structure. In addition, antibacterial activity was also observed in commercial apple juice, which showed results similar to those obtained from phosphate-buffered saline (PBS), resulting in a significant (P < 0.05) reduction for all three pathogens without any changes in color parameters (L*, a*, and b*), total phenolic compounds, and DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging activity. IMPORTANCE Photodynamic inactivation (PDI), which involves photoactivation of a photosensitizer (PS), is an emerging field of study, as it effectively reduces various kinds of microorganisms. Although there are several PSs that have been used for PDI, there is a need to find naturally occurring PSs for safer application in the food industry. Caffeic acid, a natural polyphenol found in most fruits and vegetables, has recently been studied for its potential to act as a novel photosensitizer. However, no studies have been conducted regarding its antibacterial activity depending on treatment conditions and its antibacterial mechanism. In this study, we closely examined the effectiveness of caffeic acid in combination with UV-A light for inactivating representative foodborne bacteria in liquid medium. Therefore, the results of this research are expected to be utilized as basic data for future application of caffeic acid in PDI, especially when controlling pathogens in liquid food processing.

Growth temperature influences the resistance of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium on lettuce to X-ray irradiation.

This study aimed to investigate the effect of different growth temperatures on the resistance of Escherichia coli O157:H7 and Salmonella Typhimurium to low-energy X-ray irradiation. Irradiation of contaminated phosphate-buffered saline with 0.6 kGy X-ray decreased the counts of E. coli O157:H7 cultured at 37 °C to below the detection limit (<1.0 colony-forming unit (CFU)/mL) and those of E. coli O157:H7 cultured at 25 and 15 °C by 4.82 and 4.45 log CFU/mL, respectively. The viable counts of S. Typhimurium cultured at 37, 25, and 15 °C in phosphate-buffered saline decreased by 3.56, 3.08, and 2.75 log CFU/mL, respectively, after irradiation with 0.6 kGy X-ray. Irradiation of contaminated lettuce with 0.4 kGy decreased the counts of E. coli O157:H7 cultured at 37, 25, and 15 °C by 3.97, 3.45, and 3.10 log CFU/cm2, respectively, and those of S. Typhimurium by 4.41, 3.84, and 3.40 log CFU/cm2, respectively. Growth temperature influenced pathogen resistance to X-ray irradiation by modulating cellular membrane and DNA integrity, intracellular enzyme activity, and efflux pump function. The results of this study suggest that the stress resistance status of pathogenic bacteria cultured at different growth temperatures should be considered for the application of X-ray irradiation for fresh produce sterilization.

Decontamination of dried whole black peppercorns using ultraviolet-c irradiation.

This study determined the efficacy of UV-C as a decontamination process against some foodborne bacteria in dried whole black peppercorns. Artificially-inoculated Salmonella enterica, Escherichia coli O157:H7, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus were subjected to UV-C with a surface irradiance of 0.43 mW/cm2 and were all found to exhibit a biphasic inactivation pattern with fast log-linear inactivation followed by a tail. Total log reductions (TLR) ranged from 1.92 (S. aureus) to 3.60 log CFU/g (E. coli O157:H7). Increasing the lamp number from 1 to 5 also linearly (R2 = 0.98) increased the surface irradiance from 0.43 to 1.70 and the TLR of the most resistant S. aureus from 1.92 to 2.62 log CFU/g. Quality evaluation showed very small, variable changes in color coordinates, which were not detected by a same/different test involving a 50-member sensory evaluation panel. Mercury deposition was not detected after a maximum exposure time of 90 min to 0.43 and 1.70 mW/cm2. Finally small, non-significant changes in the innate bacterial microflora of the black peppercorns were determined after 90 min-treatment using 1 lamp and 5 lamps, emphasizing the limitation of utility of UV-C as additional decontamination process for post-process-introduced microorganisms. Good Manufacturing Practices throughout the dried black peppercorn manufacturing process were recommended.

Microwave pasteurization of apple juice: Modeling the inactivation of Escherichia coli O157:H7 and Salmonella Typhimurium at 80-90 °C.

Although due to their acidity some fruit juices are considered safe, several outbreaks have been reported. For processing fruit juices, microwave heating offers advantages such as shorter come-up time, faster and uniform heating, and energy efficiency. Thus, it could be a beneficial alternative to conventional pasteurization. The objective of this study was to study the inactivation kinetics of Escherichia coli O157:H7 and Salmonella Typhimurium under microwave pasteurization at temperatures between 80 and 90 °C, i.e., at conditions that are employed in conventional pasteurization. Inoculated juices were treated at different power levels (600 W, 720 W) and treatment times (5s, 10s, 15s, 20s, 25s). Time-temperature profiles were obtained by fiber-optic sensors in contact with the samples allowing continuous data collection. The log-logistic and Arrhenius equations were used to account for the influence of the temperature history; thus, resulting in two different modeling approaches that were compared in terms of their prediction abilities. Survival kinetics including non-isothermal conditions were described by a non-linear ordinary differential equation that was numerically solved by the Runge-Kutta method (ode45 in MATLAB ®). The lsqcurvefit function (MATLAB®) was employed to estimate the corresponding survival parameters, which were obtained from freshly made apple juice, whereas the prediction ability of these parameters was evaluated on commercial apple juices. Results indicated that inactivation increased with power level, temperature, and treatment time reaching a microbial reduction up to 7 Log10 cycles. The study is relevant to the food industry because it provides a quantitative tool to predict survival characteristics of pathogens at other non-isothermal processing conditions.

Synergistic bactericidal effect and mechanism of X-ray irradiation and citric acid combination against food-borne pathogens on spinach leaves.

In this study, we investigated the antimicrobial activity of the X-ray irradiation and citric acid (CA) combination against Escherichia coli O157:H7 and Listeria monocytogenes on the surface of spinach leaves and elucidated the mechanisms underlying their synergistic interaction. Upon treatment with 0.3 kGy X-ray irradiation and 1% CA combination, the cell counts of E. coli O157:H7 and L. monocytogenes reduced by 4.23 and 3.69 log CFU/mL on spinach leaves, respectively. The synergistic reduction in the cell counts of E. coli O157:H7 and L. monocytogenes by the combination treatment was 0.95 and 1.14 log units, respectively. The X-ray and CA combination exerts its antimicrobial effect by damaging the bacterial cell membrane and enhancing the generation of intracellular reactive oxygen species in the pathogens. The enhanced bactericidal effect of the combination treatment may not be due to the loss of intracellular enzyme activity. We also evaluated the effect of the combination treatment on the quality attributes of spinach leaves. The combination treatment did not result in adverse changes in color and texture of spinach leaves. These results demonstrate the potential of citric acid and X-ray irradiation combination for decontaminating foodborne pathogens on fresh produce.

Bactericidal and synergistic effects of X-ray irradiation and gallic acid against foodborne pathogens on lettuce.

The objectives of this study were to evaluate the bactericidal effects of X-ray irradiation and gallic acid (GA) against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on lettuce leaves and in phosphate-buffered saline (PBS). Inoculated PBS and lettuce were exposed to X-rays (0.05, 0.1, and 0.15; 0.1, 0.2, and 0.3 kGy, respectively), and GA was applied to lettuce leaves as a solution and in PBS at concentrations of 0.5% (w/v). Combined treatment with 0.3 kGy and 0.5% GA reduced E. coli O157:H7, S. Typhimurium, and L. monocytogenes cell counts 5.41, 2.57, and 1.36 log CFU/cm2 on lettuce, respectively. Combined treatment with 0.15 kGy X-ray and 0.5% GA reduced counts for the same species by 6.54, 4.24, and 1.51 log CFU/mL in PBS. The combined treatments exerted a synergistic antibacterial effect against E. coli O157:H7 on lettuce, but not against S. Typhimurium or L. monocytogenes. In PBS, the synergistic effect was confirmed in both E. coli O157:H7 and S. Typhimurium cells. Mechanistic investigations indicated that the synergistic antibacterial effect was associated with intracellular reactive oxygen species (ROS) generation and bacterial cell membrane damage. Additionally, the X-ray and GA combination treatment did not adversely affect the color, total phenol content, and texture of lettuce. These findings demonstrate that treatment with X-ray radiation and GA can enhance the microbiological safety of fresh produce.

Inactivating foodborne pathogens in apple juice by combined treatment with fumaric acid and ultraviolet-A light, and mechanisms of their synergistic bactericidal action.

We evaluated the bactericidal efficacy of the simultaneous application of ultraviolet-A (UV-A) irradiation and fumaric acid (FA) against Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes in apple juice and as well as investigated the effects of this treatment on product quality. Further, we elucidated the mechanisms underlying their synergistic bactericidal action. Simultaneous UV-A light irradiation and 0.1% FA treatment for 30 min resulted in 6.65-, 6.27-, and 6.49-log CFU/ml reductions in E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, which involved 3.15, 2.21, and 3.43 log CFU reductions, respectively, and these were attributed to the synergistic action of the combined treatments. Mechanistic investigations suggested that the combined UVA-FA treatment resulted in significantly greater bacterial cell membrane damage and intracellular reactive oxygen species (ROS) generation. UVA-FA treatment for 30 min did not cause significant changes to the color, nonenzymatic browning index, pH, and total phenolic content of apple juice. These results suggest that combined UVA-FA treatment can be effectively used to control foodborne pathogens in apple juice without affecting its quality.

The Synergistic Bactericidal Mechanism of Simultaneous Treatment with a 222-Nanometer Krypton-Chlorine Excilamp and a 254-Nanometer Low-Pressure Mercury Lamp.

The purpose of this study was to investigate the synergistic bactericidal effect of 222-nm KrCl excilamp and 254-nm low-pressure (LP) Hg lamp simultaneous treatment against Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium, and Listeria monocytogenes in tap water and to identify the synergistic bactericidal mechanism. Sterilized tap water inoculated with pathogens was treated individually or simultaneously with a 254-nm LP Hg lamp or 222-nm KrCl excilamp. Overall, for all pathogens, an additional reduction was found compared to the sum of the log unit reductions of the individual treatments resulting from synergy in the simultaneous treatment with both kinds of lamps. In order to identify the mechanism of this synergistic bactericidal action, the form and cause of membrane damage were analyzed. Total reactive oxygen species (ROS) and superoxide generation as well as the activity of ROS defense enzymes then were measured, and the overall mechanism was described as follows. When the 222-nm KrCl excilamp and the 254-nm LP Hg lamp were treated simultaneously, inactivation of ROS defense enzymes by the 222-nm KrCl excilamp induced additional ROS generation following exposure to 254-nm LP Hg lamp (synergistic) generation, resulting in synergistic lipid peroxidation in the cell membrane. As a result, there was a synergistic increase in cell membrane permeability leading to a synergistic bactericidal effect. This identification of the fundamental mechanism of the combined disinfection system of the 222-nm KrCl excilamp and 254-nm LP Hg lamp, which exhibited a synergistic bactericidal effect, can provide important baseline data for further related studies or industrial applications in the future.IMPORTANCE Contamination of pathogenic microorganisms in water plays an important role in inducing outbreaks of food-borne illness by causing cross-contamination in foods. Thus, proper disinfection of water before use in food production is essential to prevent outbreaks of food-borne illness. As technologies capable of selecting UV radiation wavelengths (such as UV-LEDs and excilamps) have been developed, wavelength combination treatment with UV radiation, which is widely used in water disinfection systems, is actively being studied. In this regard, we have confirmed synergistic bactericidal effects in combination with 222-nm and 254-nm wavelengths and have identified mechanisms for this. This study clearly analyzed the mechanism of synergistic bactericidal effect by wavelength combination treatment, which has not been attempted in other studies. Therefore, it is also expected that these results will play an important role as baseline data for future research on, as well as industrial applications for, the disinfection strategy of effective wavelength combinations.

Decontamination Effect of the Spindle and 222-Nanometer Krypton-Chlorine Excimer Lamp Combination against Pathogens on Apples (Malus domestica Borkh.) and Bell Peppers (Capsicum annuum L.).

In this study, we developed a washing system capable of decontaminating fresh produce by combining the Spindle apparatus, which detaches microorganisms on sample surfaces, and a 222-nm krypton-chlorine excimer lamp (KrCl excilamp) (Sp-Ex) and investigated their decontamination effect against Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes on apple (Malus domestica Borkh.) and bell pepper (Capsicum annuum L.) surfaces. Initial levels of the three pathogens were approximately 108 CFU/sample. Both E. coli O157:H7 and S. Typhimurium were reduced to below the detection limit (2.0 log CFU/sample) after 5 and 7 min of treatment on apple and bell pepper surfaces, respectively. The amounts of L. monocytogenes on apple and bell pepper surfaces were reduced by 4.26 and 5.48 logs, respectively, after 7 min of treatment. The decontamination effect of the Sp-Ex was influenced by the hydrophobicity of the sample surface as well as the microbial cell surface, and the decontamination effect decreased as the two hydrophobicity values increased. To improve the decontamination effect of the Sp-Ex, Tween 20, a surfactant that weakens the hydrophobic interaction between the sample surface and pathogenic bacteria, was incorporated into Sp-Ex processing. It was found that its decontamination effect was significantly (P < 0.05) increased by the addition of 0.1% Tween 20. Sp-Ex did not cause significant quality changes in apple or bell pepper surfaces during 7 days storage following treatment (P > 0.05). Our results suggest that Sp-Ex could be applied as a system to control pathogens in place of chemical sanitizer washing by the fresh-produce industry.IMPORTANCE Although most fresh-produce processing currently controls pathogens by means of washing with sanitizers, there are still problems such as the generation of harmful substances and changes in product quality. A combination system composed of the Spindle and a 222-nm KrCl excilamp (Sp-Ex) developed in this study reduced pathogens on apple and bell pepper surfaces using sanitizer-free water without altering produce color and texture. This study demonstrates the potential of the Sp-Ex to replace conventional washing with sanitizers, and it can be used as baseline data for practical application by industry. In addition, implementation of the Sp-Ex developed in this study is expected not only to meet consumer preference for fresh, minimally processed produce but also to reduce human exposure to harmful chemicals while being beneficial to the environment.

Increased Resistance of Salmonella enterica Serovar Typhimurium and Escherichia coli O157:H7 to 222-Nanometer Krypton-Chlorine Excilamp Treatment by Acid Adaptation.

In this study, we examined the change in resistance of Salmonella enterica serovar Typhimurium and Escherichia coli O157:H7 to 222-nm krypton-chlorine (KrCl) excilamp treatment as influenced by acid adaptation and identified a mechanism of resistance change. In addition, we measured changes in apple juice quality indicators, such as color, total phenols, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, during treatment. Non-acid-adapted and acid-adapted pathogens were induced by growing the cells in tryptic soy broth without dextrose (TSB w/o D) at pH 7.3 and in TSB w/o D at pH 5.0 (adjusted with HCl), respectively. For the KrCl excilamp treatment, acid-adapted pathogens exhibited significantly (P < 0.05) higher D5d values, which indicate dosages required to achieve a 5-log reduction, than those for non-acid-adapted pathogens in both commercially clarified apple juice and phosphate-buffered saline (PBS), and the pathogens in the juice showed significantly (P < 0.05) higher D5d values than those for pathogens in PBS because of the UV-absorbing characteristics of apple juice. Through mechanism identification, it was found that the generation of lipid peroxidation in the cell membrane, inducing cell membrane destruction, was significantly (P < 0.05) lower in acid-adapted cells than in non-acid-adapted cells for the same amount of reactive oxygen species (ROS) generated at the same dose because the ratio of unsaturated to saturated fatty acids (USFA/SFA) in the cell membrane was significantly (P < 0.05) decreased as a result of acid adaptation. Treated apple juice showed no significant (P > 0.05) difference in quality indicators compared to those of untreated controls during treatment at 1,773 mJ/cm2IMPORTANCE There is a need for novel, mercury-free UV lamp technology to replace germicidal lamps containing harmful mercury, which are routinely utilized for UV pasteurization of apple juice. In addition, consideration of the changes in response to antimicrobial treatments that may occur when pathogens are adapted to the acid in an apple juice matrix is critical to the practical application of this technology. Based on this, an investigation using 222-nm KrCl excilamp technology, an attractive alternative to mercury lamps, was conducted. Our study demonstrated increased resistance to 222-nm KrCl excilamp treatment as pathogens adapted to acids, and this was due to changes in reactivity to ROS with changes in the fatty acid composition of the cell membrane. Despite increased resistance, the 222-nm KrCl excilamp achieved pathogen reductions of 5 log or more at laboratory scale without affecting apple juice quality. These results provide valuable baseline data for application of 222-nm KrCl excilamps in the apple juice industry.

Effect of gamma irradiation on inactivation of Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes on pistachios.

This study evaluated the efficacy of gamma irradiation to inactivate food-borne pathogens on pistachios (Pistacia vera L.). Pistachios inoculated with Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes were subjected to gamma irradiation in the range of 0, 0·5, 1, 3 and 5 kGy, and colour change was evaluated after treatment and during storage at room temperature. Pathogen populations decreased with increasing treatment doses. A gamma irradiation dose of 5 kGy decreased the three pathogens on pistachios to under the detection limit (1·0 log CFU per g) without effecting colour change. During storage following treatment, pathogens were reduced due to the postirradiation effect. D-values of pathogens on pistachios showed that L. monocytogenes was more resistant to gamma irradiation than was E. coli O157:H7 or S. Typhimurium. During gamma irradiation treatment, L, a and b values of pistachios did not significantly change but these values changed during storage. These results show that gamma irradiation has potential as a nonthermal process for inactivating food-borne pathogens in pistachios without inducing colour changes. SIGNIFICANCE AND IMPACT OF THE STUDY: Generally, nuts have lower water activity which precludes the growth of food-borne pathogens. But, food-borne outbreaks due to pathogen-contaminated nuts have been reported in the last few decades. Pistachios are one of the most popular nuts and have many health beneficial effects. However, many pasteurization interventions have been used to reduce pathogens on pistachios, but most of them are not effective. This study confirms the effectiveness of gamma irradiation on pasteurization of pistachios. This may be helpful in nut processing industries to ensure the microbial safety of nuts.

Effects of pulsed light and sanitizer wash combination on inactivation of Escherichia coli O157:H7, microbial loads and apparent quality of spinach leaves.

The purpose of this study was to investigate the efficacy of pulsed light (PL), a new formula of sanitizer (HEN) consisting of hydrogen peroxide, EDTA and Nisin, as well as synergy of PL and HEN sanitizer (PL-HEN) wash in inactivating E. coli O157:H7 on spinach. The treatment effect on microbial loads and apparent quality during 13 days storage at 4 °C was also determined. A bacterial cocktail containing three strains of E. coli O157:H7 was used as inoculum based on their association with produce-related outbreaks. Spinach leaves were spot inoculated on surface before treating with PL (1-63J/cm2), HEN sanitizer wash (2 min) or their combinations. PL inactivation was influenced significantly at low doses. Treatment dose of 15.75 J/cm2, equivalent to 15 s intense PL treatment, was found optimal above which adverse quality effect was evident. The optimal PL dose resulted 2.7 log CFU/g reduction of E. coli O157:H7 while a rapid 2 min wash in sanitizer formulation HEN, provided comparatively low, 1.8 log CFU/g, reduction of the pathogen. Two different sequences of PL and HEN treatment combinations were tested. In PL-HEN treatment, inoculated leaves were first treated at optimal PL dose (15.75 J/cm2) followed by 2 min immersion in HEN whereas in HEN-PL treatment, leaves were first washed in HEN before PL exposure. HEN-PL treatment indicated a compound inactivation activity (4.6 logs reduction) while PL-HEN treatment indicated a strong synergistic inactivation as E. coli cells were not detectable after treatment indicating >5 log reduction. The PL-HEN treatment not only significantly reduced spoilage microbial populations on spinach but also slowed their growth during storage. Furthermore, the visual and firmness quality of spinach were not significantly affected by the PL-HEN treatment. Overall, our results demonstrate that integrated PL-HEN technology can be used to enhance microbial safety of spinach.

Effect of 222-nm krypton-chloride excilamp treatment on inactivation of Escherichia coli O157:H7 and Salmonella Typhimurium on alfalfa seeds and seed germination.

We examined the control effect of a 222-nm KrCl excilamp on foodborne pathogens on alfalfa seeds and compared it with a conventional 254-nm low-pressure (LP) Hg lamp. When the 222-nm KrCl excilamp treated seeds at 87, 174 and 261 mJ/cm2, the log reductions of Escherichia coli O157:H7 (E. coli O157:H7) were 0.85, 1.77, and 2.77, respectively, and Salmonella Typhimurium (S. Typhimurium) experienced log reductions of 1.22, 2.27, and 3.04, respectively. When the 254-nm LP Hg lamp was applied at 87, 174, and 261 mJ/cm2, the log reductions of E. coli O157: H7 were 0.7, 1.16, and 1.43, respectively, and those of S. Typhimurium were 0.75, 1.15, and 1.85, respectively. Therefore, it was shown that the 222-nm KrCl excilamp was more effective than the 254-nm LP Hg lamp in reducing foodborne pathogens. The germination rate decreased to less than 80% after 261 mJ/cm2 treatment with the 254-nm LP Hg lamp, while more than 90% was maintained with 261 mJ/cm2 222-nm KrCl excilamp treatment. DNA damage assay showed that the difference in germination rate was due to DNA damage resulting from 254-nm LP Hg lamp treatment. However, 222 nm KrCl excilamp treatment did not cause DNA damage, resulting in no difference in germination rate compared to that of non-treated alfalfa seeds. Overall, these results demonstrate the utility of the 222-nm KrCl excilamp as a foodborne pathogen control intervention for the alfalfa seed industry.

Inactivation of Escherichia coli O157 and Salmonella Enteritidis in raw beef liver by gamma irradiation.

Irradiation of ground beef and beef liver inoculated with Escherichia coli O157 466 and DT66 and Salmonella Enteritidis 3313 were performed with gamma rays from cobalt-60 at refrigerated and frozen temperatures under air- and vacuum-packaged conditions. Results showed that D10 values for all pathogens in frozen beef liver were higher than those in frozen ground beef samples, with significant differences observed between the D10 values of E. coli O157 466 and S. Enteritidis 3313 under air-packaged conditions, as well as in E. coli O157 DT66 and S. Enteritidis 3313 under vacuum-packaged conditions. To verify effective bacterial inactivation under high bacterial-contamination levels (105-107 CFU/g), survival/death interfaces of E. coli O157 DT66 and S. Enteritidis 3313 inoculated in beef liver under vacuum-packaged and frozen conditions were constructed, with results suggesting that doses from 5.3 kGy to 5.5 kGy and 8.2 kGy-8.5 kGy would be sufficient to kill 105 CFU/g of E. coli O157 and S. Enteritidis 3313, respectively, at a 95%-99% predicted confidence interval. These results suggested that food matrixes containing high amounts of antioxidants (such as beef liver) and treated under frozen and vacuum-packaged conditions require additional consideration and evaluation for applications of irradiation treatment.

Ultrasound treatment combined with fumaric acid for inactivating food-borne pathogens in apple juice and its mechanisms.

The purpose of this study was to evaluate the synergistic bactericidal efficacy of combining ultrasound (US) and fumaric acid (FA) treatment against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in apple juice and to identify the synergistic bactericidal mechanisms. Additionally, the effect of combination treatment on juice quality was determined by measuring the changes in color, pH, non-enzymatic browning index, and total phenolic content. A mixed cocktail of the three pathogens was inoculated into apple juice, followed by treatment with US (40 kHz) alone, FA (0.05, 0.1, and 0.15%) alone, and a combination of US and FA for 1, 2, 3, 4, and 5 min. Combined US and 0.15% FA treatment for 5 min achieved 5.67, 6.35, and 3.47 log reductions in E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, with the 1.55, 2.37, and 0.57 log CFU reductions attributed to the synergistic effect. Although the pH value slightly decreased as FA increased, there were no significant (P > 0.05) differences in color values, browning indices, and phenolic content between untreated and treated samples. To identify the mechanism of this synergistic bactericidal action, membrane integrity, malfunctions in the membrane efflux pump, and intracellular enzyme activity were measured. The analyses confirmed that damage to the cell envelope (membrane integrity and efflux pump) was strongly related to the synergistic microbial inactivation. These results suggest that simultaneous application of US treatment and FA is a novel method for ensuring the microbial safety of apple juice.

Elevated Inactivation Efficacy of a Pulsed UVC Light-Emitting Diode System for Foodborne Pathogens on Selective Media and Food Surfaces.

UVC light, a strong surface disinfection technology, is used worldwide to ensure not only environmental safety but also food safety. Several drawbacks associated with the use of mercury-containing UV lamps, especially human and environmental health risks, led to the Minamata Convention on Mercury, which prohibits the manufacture and import/export of products containing mercury. Therefore, light-emitting diode (LED)-based UVC irradiation, a new technology that is ecofriendly and represents an effective UV light source, has been researched recently. To date, however, there has been no report describing pulsed UVC-LED irradiation for improvement of inactivation of foodborne pathogens, although much research regarding conventional pulsed xenon lamps has been published. In this investigation, we evaluated the enhanced bactericidal effect of a pulsed UVC-LED system, compared to continuous irradiation, and optimum conditions for maximizing the effect were determined. Also, the differences in inactivation between pulsed and continuous UVC-LED irradiation were determined by inactivation mechanism analyses. The combination of 20-Hz frequency and 50% duty ratio for pulsed UVC-LED irradiation achieved 4- to 5-log-unit reductions of Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes; this combination showed the greatest bactericidal effect among various treatment conditions using 2 or 5 mJ/cm2 In mechanism assessments, membrane integrity (propidium iodide uptake) was not affected by UVC-LED treatment but membrane potential [bis-(1,3-dibutylbarbituric acid)trimethine oxonol [DiBAC4(3)] accumulation] showed significantly different values when pulsed and continuous treatments were compared. Changes in membrane lipid peroxidation and respiratory enzyme activity were attributed to generation of more reactive oxygen species by pulsed UVC-LED irradiation.IMPORTANCE In 2013, the United Nations Environment Programme convened the Minamata Convention on Mercury, which prohibits trade in mercury-containing products in order to ensure human health. It will be effectuated in 2020; use of low-pressure mercury lamps will be discontinued and a new UV light source selected to replace the conventional technology. In this regard, UVC-LEDs have been developed and the fundamental inactivating effect has been researched. However, a pulsed UVC-LED system has not been studied, because of the difficulty of generating a UVC-LED pulse wave. An optical chopper system that physically divides the light with an adjustable blade, with personalized frequency and duty ratio settings, was introduced for generation of pulsed UVC-LED irradiation. This study elucidated the efficacy of a pulsed UVC-LED system and investigated its enhanced bactericidal effect in mechanism analyses.

UVC LED Irradiation Effectively Inactivates Aerosolized Viruses, Bacteria, and Fungi in a Chamber-Type Air Disinfection System.

In this study, the possibility of inactivating viral, bacterial, and fungal aerosols in a chamber-type air disinfection system by using a UVC light-emitting-diode (LED) array was investigated and inactivation rate constants of each microorganism were calculated in fitting curves of surviving populations. UVC LED array treatment effectively inactivated viral infectivity, achieving 5-log reductions within 45 mJ/cm2 for MS2, Qß, and ϕX174 viruses. UVC LED array effectiveness in inactivating Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Listeria monocytogenes, and Staphylococcus aureus aerosols achieved 2.5- to 4-log reductions within 1.5 to 4.6 mJ/cm2 Also, 4-log reductions of Aspergillus flavus and Alternaria japonica were achieved at a dosage of 23 mJ/cm2 using UVC LED array irradiation. The highest UV susceptibility, represented by the inactivation rate constant, was calculated for bacteria, followed by fungi and viruses. UVC LED, an innovative technology, can effectively inactivate microorganisms regardless of taxonomic classification and can sufficiently substitute for conventional mercury UV lamps.IMPORTANCE The United Nations Environment Programme (UNEP) convened the Minamata Convention on Mercury in 2013 to ban mercury-containing products in order to ensure human and environmental health. It will be effectuated in 2020 to discontinue use of low-pressure mercury lamps and new UV-emitting sources have to replace this conventional technology. However, the UV germicidal irradiation (UVGI) system still uses conventional UV lamps, and no research has been conducted for air disinfection using UVC LEDs. The research reported here investigated the inactivation effect of aerosolized microorganisms, including viruses, bacteria, and fungi, with an UVC LED module. The results can be utilized as a primary database to replace conventional UV lamps with UVC LEDs, a novel type of UV emitter. Implementation of UVC LED technology is truly expected to significantly reduce the extent of global mercury contamination, and this study provides important baseline data to help ensure a healthier environment and increased health for humanity.

Novel sanitization approach based on synergistic action of UV-A light and benzoic acid: Inactivation mechanism and a potential application in washing fresh produce.

Antimicrobial activity of the simultaneous UV-A light and benzoic acid (BA) treatment against stationary phase Escherichia coli O157:H7 was investigated. While 15 mM BA or UV-A light exposure for 30 min alone caused < 1 logarithmic reduction in the bacterial population, > 5 logarithmic reductions were induced by the simultaneous application of UV-A and 15 mM BA in 30 min, demonstrating a synergistic antimicrobial effect. Due to its ability to increase cell membrane permeability, addition of EDTA (1 mM) was able to decrease the required concentration of BA in the simultaneous treatment from 15 to 8 mM. Microbial inactivation was a result of simultaneous membrane damage, intracellular acidification, and intracellular oxidative stress. The simultaneous treatment was effective in the presence of organic load of up to 500 mg/L of chemical oxygen demand (COD) and was able to lower cross-contamination risk during simulated washing of spinach (Spinacia oleracea) without adversely affecting its color.

Fog, phenolic acids and UV-A light irradiation: A new antimicrobial treatment for decontamination of fresh produce.

This study evaluates synergistic interactions of food grade phenolic acids (gallic and ferulic acid) and UV-A light to achieve decontamination of fresh produce using a fog to improve dispersion of the phenolic acids on produce surface. Nonvirulent strains of Escherichia coli O157:H7 and Listeria innocua were used as model bacteria and spinach was selected as a model fresh produce. Synergistic combination of a fog deposited phenolic acid and a UV-A light treatment achieved reduction in bacterial plate count up to 2 log CFU/cm2 independently of the initial load of the bacteria (104 or 106 CFU/cm2). Following the treatment, fog deposited gallic and ferulic acid could be easily removed from the surface of produce by immersion in water and the treatment did not significantly alter the total endogenous phenolic content of spinach. The treatment also did not affect the texture, but impacted the color of the spinach leaves on a Hunter's Lab scale although the visual color changes were small. Overall, this technology may aid in developing alternative approaches for decontamination processes using food grade compounds.