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Antibacterial packaging holds promise in addressing food spoilage by inactivating bacteria, but current antimicrobial packaging solutions face challenges like depletion of antibacterials and concerns of antibiotic abuse. In response to these limitations of existing packaging materials, we developed a novel antibacterial packaging film by incorporating titanium dioxide (TiO 2 )- tetra(4-carboxyphenyl) porphyrin (TcPP) conjugates into cellulose nanofibrils (CNF) films. Unlike conventional antimicrobial packaging, this film harnesses visible light energy to excite electrons from TcPP to TiO 2 , generating reactive oxygen species (ROS) that inactivate bacteria without relying on antibiotics. Results demonstrated that the film reduced 4.5, 4.6, 4.1, and 4.7-log Escherichia coli, Pseudomonas fluorescens, Leuconostoc lactis, and Listeria innocua, respectively, in phosphate-buffered saline within 72 h under 6000 lux light (3.13 mW/cm 2 ). The antimicrobial efficacy decreased as the light intensity decreased. Notably, it retains significant antimicrobial properties even under an extremely low light intensity of 600 lux (0.60 mW/cm 2 ). The analysis also revealed that singlet oxygen and hydrogen peroxide are the major generated ROS from the film under light exposure. When applied to cucumbers, the film reduced E. coli by 3.5 logs after 48-hour light exposure. The designed photocatalytic antibacterial film represents a major advancement in sustainable food preservation, reducing food waste by extending the shelf life of fresh produce., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Haibo Huang has patent #Photo-catalytic antimicrobial packaging films. Application No. 63/358,322 pending to Haibo Huang. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In this study the suitability of a thin-film reactor (TFR) equipped with special flow guiding elements (FGE) was examined to analyse its capability to inactivate microorganisms in milk. Experiments were carried out with UHT-milk inoculated with Escherichia coli (E. coli), DH5α and Listeria innocua (L. innocua) WS 2258. Furthermore, the inactivation of microorganisms originally occurring in raw milk was investigated. E. coli, DH5α and L. innocua serving as biodosimeter were reduced by 4.58-log and 3.19-log, respectively. In milk, the original microorganisms showed a 4-log reduction. Without FGE the reduction was below 0.13-log. Thus, it can be derived that the efficacy of a UV-C thin-film reactor processing absorptive media like milk can be highly improved using FGE., (Copyright © 2021 Elsevier B.V. All rights reserved.)

We aimed to study the efficacy of a water-assisted UVC light device (WUVC) as an innovative clean technology for the disinfection of fresh sound tomatoes and processing wash water and water turbidity was evaluated as a critical parameter. First, wash waters with different turbidities (from 0.4 to 828 NTU) were inoculated with Listeria innocua and treated in the WUVC device at different dosages. Secondly, fresh tomatoes, inoculated with L. innocua and non-inoculated ones, were treated using the WUVC device containing wash water of different turbidities for different times. The reduction of L. innocua populations on wash water and on the surface of tomato was influenced by turbidity; lower reduction values were observed at higher turbidities. Washing tomatoes with tap water with UVC lamps off (control treatment, TW) decreased L. innocua population on the surface of tomatoes but did not eliminate those bacteria that went into the water. Contrarily, when UVC lights were on, L. innocua population in wash water after treatment significantly decreased, those in clean water being the lowest populations. Reductions of native microbiota on the clean water treated with the highest UV-C radiation dose were lower than those obtained when tomatoes were artificially inoculated. We demonstrated that high reductions of L. innocua population on fresh tomatoes could be achieved using the WUVC system but some drawbacks related to the increase of turbidity should be solved for its implementation in real conditions., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Highlights: Thermal and nonthermal methods can support a 5-log CFU reduction of model bacteria introduced into tiger nut milk. Thermal treatment of tiger nut milk results in significant loss of protein, antioxidants, and quality properties. HHP or UV-C treatment of tiger nut milk retains quality and nutritional characteristics. HHP or UV-C are suitable for the pasteurization of tiger nut milk.

There is a critical unmet need to improve microbial safety of fresh fruits and vegetables. Current sanitation approaches cannot achieve >2 log inactivation of bacteria on fresh produce. Thus, there is a need to develop antimicrobial strategies that can consistently achieve >2 logs of bacterial inactivation on the surface of diverse fresh produce. Furthermore it is highly desired that these antimicrobial strategies have reduced environmental impact and are clean label solutions for food products. In this study, we evaluated the combination of curcumin and UV-A light radiation for the inactivation of inoculated E. coli O157:H7 and L. innocua bacterial cells on the surface of spinach, lettuce and tomatoes. Curcumin was deposited on the surface of fresh produce by either aerosolization or conventional spray-atomization methods before exposing the contaminated produce to UV-A light for 10 min (total light fluence of 20.4 kJ m -2 ). Results showed that the proposed combination of aerosolized or sprayed curcumin and UV-A light radiation can reduce the initial Escherichia coli O157:H7 and Listeria innocua load from 6 log CFU cm -2 to approximately 3 log CFU cm -2 on spinach, lettuce and tomato surfaces. Furthermore, there was no significant difference in bacterial reduction between the different types of inoculated fresh produce surfaces (P > .05). Interestingly, subsequent curcumin deposition and UV-A light exposure cycles were not able to further reduce the bacterial load below the observed threshold of approximately 3 log CFU cm -2 . Lastly, the combination of aerosolized curcumin and UV-A light radiation did not affect the color or the texture of the treated fresh produce samples. The findings described in this study illustrate the potential of applying aerosolized or sprayed curcumin under UV-A light illumination to improve microbial safety of fresh produce products., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The effect of pulsed light treatment on the lag phase and the maximum specific growth rate of Listeria innocua was determined in culture media at 7 °C. Fluences of 0.175, 0.350 and 0.525 J/cm 2 were tested. The lag phase of the survivors increased as fluence did, showing significant differences for all the doses; an 8.7-fold increase was observed at 0.525 J/cm 2 . Pulsed light decreased the maximum specific growth rate by 38% at the same fluence. Both parameters were also determined by time-lapse microscopy at 25 °C in survivors to 0.525 J/cm 2 , with an increase of 13-fold of the lag phase and a 45% decrease of the maximum specific growth rate. The higher the fluence, the higher the variability of both parameters was. To characterize pulsed light damage on L. innocua, the formation of dimers on DNA was assessed, and a proteomic study was undertaken. In cells treated with 0.525 J/cm 2 , cyclobutane pyrimidine dimers and pyrimidine (6-4) pyrimidone photoproducts were detected at 5:1 ratio. Pulsed light induced the expression of three proteins, among them the general stress protein Ctc. Furthermore, treated cells showed an up-regulation of proteins related to metabolism of nucleotides and fatty acids, as well as with translation processes, whereas flagellin and some glucose metabolism proteins were down-regulated. Differences in the proteome of the survivors could contribute to explain the mechanisms of adaptation of L. innocua after pulsed light treatment., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study investigated the efficacy of power ultrasound (US) for the inactivation of Escherichia coli and Listeria innocua in the presence of sodium salt and salt replacers. Inoculated bacteria suspensions were treated at ultrasonic frequencies of 33 or 20 kHz alone or in combination, and in the presence of 5% NaCl, 5% KCl or 5% NaCl/KCl. Inactivation curves were fitted to the Weibull and the Biphasic models. The goodness of the fit for each model was evaluated based on R 2 and RMSE, while AIC and BIC values were used to choose the best model predictor. The Weibull and the biphasic models showed high regression coefficient (R 2  > 0.99) and low RMSE (<0.03) values. According to the results, inactivation up to 6 log for E. coli K12 and to 4 log for L. innocua could be achieved within one hour of ultrasound treatment. However, the presence of NaCl, or its substitution with KCl did not affect the degree of inhibition for both microorganisms. The results of this study suggest that power ultrasound treatment may be employed for the inactivation of microorganisms when low salt or salt substitutes are employed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The antimicrobial efficacy of 400 nm photoirradiated caffeic acid (CA, 5 mM) was evaluated against Escherichia coli O157:H7 and Listeria innocua. A stronger antimicrobial effect was observed on E. coli than on L. innocua where the combined treatment resulted in 4 and 1 log(CFU/mL) reductions, respectively. The treatment's effects on cellular metabolism (resazurin assay), uptake of CA (fluorescence technique) and membrane damage (propidium iodide assay) were studied in both species. CA uptake increased in both species, but membrane damage was only observed in E. coli O157:H7. The treatment had minimal impact on metabolic activity in both species. The treatment applied to the surface of spinach leaves was found to be effective against E. coli O157:H7. The novel treatment proposed in this study has the potential to improve the microbial food safety of fresh produce.

The synergistic combination of the food colorant Erythrosin B (E-B, FD&C 3) (0, 25, and 50μM) and low-frequency ultrasound (20kHz, 0.86-0.90WmL -1 ) was evaluated against Listeria innocua. Although E-B was antibacterial by itself, the inactivation rate significantly increased in a concentration-dependent manner upon exposure to ultrasound and followed a sigmoidal behavior. The enhanced antimicrobial effect of E-B in the presence of ultrasound can be explained in part from a microbubble disappearance study in which it was confirmed that the presence of E-B enhances inertial cavitation, thereby enhancing the antimicrobial effect of ultrasound. The inactivation rate in a sequential treatment, where L. innocua was sonicated for 4min followed by exposure to 25μM Erythrosin B, was comparable to that obtained by the simultaneous treatment, indicating complementary mechanisms of inactivation. Fluorescence microscopy showed attachment of E-B to the cells, which may explain its intrinsic antimicrobial property. Other mechanism may include the confirmed decrease in the cavitation threshold of water by addition of E-B, resulting in more effective cavitation. The study offers a proof-of-concept of a novel approach to complement ultrasound treatment for enhanced microbial inactivation., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Aims: Pulsed light (PL) is an upcoming nonthermal decontamination technology mainly used for surface sterilization. The objective of this study was to investigate the extent of cellular damage caused by PL treatments of Listeria innocua and Escherichia coli on a polysaccharide surface in order to gain knowledge about the main inactivation pathways., Methods and Results: The impact of PL on the cellular ATP level was investigated as well as the bacterial ability to take up fluorescently labelled glucose (2-NBDG). Furthermore, the extent of DNA damages was assessed by qPCR. The ability of L. innocua and E. coli to photorepair under artificial daylight exposure was quantified. Finally, the induction of reactive oxygen species (ROS) and lipid peroxidation were studied by fluorometric detection of ROS and thiobarbituric acid reactive substances (TBARS). It is shown that intracellular ATP levels and glucose uptake ability do not correlate with the immediate loss of bacterial reproducibility, which indicates that cellular activity and energy may remain on a relatively high level, although growth on tryptic soy agar is not observable. Sequence specific investigation of PL induced DNA damages by qPCR revealed distinct differences between L. innocua and E. coli although the observed inactivation efficacy of PL by the culture based method was similar. Photoreactivation has been observed for both bacteria, a higher recovery rate of up to 2 log was seen in case of E. coli. Intracellular ROS and lipid peroxides were both detectable at relatively high fluencies with E. coli so the contribution of oxidative damage to microbial inactivation of PL cannot be excluded., Conclusions: Escherichia coli as well as L. innocua cells have proven to maintain residual cellular activity after having been exposed to PL even when they are not able to reproduce any more. High proportions of sublethal damages were also obvious with regard to occurring photoreactivation. The destruction of bacterial DNA seems to be the primary mechanism of inactivation of PL but the involvement of other factors like oxidative stress cannot be excluded., Significance and Impact of the Study: The observed data underline that bacteria are not immediately inactivated after exposure to PL as different indicators of cellular energy are still detectable even when cells do not reproduce on solid media any more. DNA is the primary target of PL, but as the extent of damage among different bacteria may not reveal their actual sensitivity, other destructive effects should also be considered., (© 2015 The Society for Applied Microbiology.)

Cheese products are susceptible to postprocessing cross-contamination by bacterial surface contamination during slicing, handling, or packaging, which can lead to food safety issues and significant losses due to spoilage. This study examined the effectiveness of pulsed-light (PL) treatment on the inactivation of the spoilage microorganism Pseudomonas fluorescens, the nonenterohemorrhagic Escherichia coli ATCC 25922 (nonpathogenic surrogate of Escherichia coli O157:H7), and Listeria innocua (nonpathogenic surrogate of Listeria monocytogenes) on cheese surface. The effects of inoculum level and cheese surface topography and the presence of clear polyethylene packaging were evaluated in a full factorial experimental design. The challenge microorganisms were grown to early stationary phase and subsequently diluted to reach initial inoculum levels of either 5 or 7 log cfu/slice. White Cheddar and process cheeses were cut into 2.5×5 cm slices, which were spot-inoculated with 100 µL of bacterial suspension. Inoculated cheese samples were exposed to PL doses of 1.02 to 12.29 J/cm(2). Recovered survivors were enumerated by standard plate counting or the most probable number technique, as appropriate. The PL treatments were performed in triplicate and data were analyzed using a general linear model. Listeria innocua was the least sensitive to PL treatment, with a maximum inactivation level of 3.37±0.2 log, followed by P. fluorescens, with a maximum inactivation of 3.74±0.8 log. Escherichia coli was the most sensitive to PL, with a maximum reduction of 5.41±0.1 log. All PL inactivation curves were nonlinear, and inactivation reached a plateau after 3 pulses (3.07 J/cm(2)). The PL treatments through UV-transparent packaging and without packaging consistently resulted in similar inactivation levels. This study demonstrates that PL has strong potential for decontamination of the cheese surface., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Inactivation of Listeria innocua by pulsed light (PL) was evaluated at different post-treatment temperature and illumination conditions. The impact of post-PL-treatment temperature on L. innocua culturability was evaluated for cells cultured at 37 °C (optimal growth temperature) and 4 °C (classical refrigerated food temperature). For both culture conditions, significant higher reductions (up to 3 log) were observed after post-PL-treatment temperature of 4 °C than of 37 °C. Contrarily, L. innocua culturability after PL treatment increased up to 2.2 log in presence of daylight illumination in comparison to dark storage. This photorepair mechanism was quickly activated reaching the maximum photoreactivation level after only 30 min of illumination. Moreover, photorepair capacity was rapidly reduced by increasing the time in darkness from PL treatment to samples illumination, being completely lost after time in darkness equal or greater than 5 h. According to these findings, the combination of PL with post-treatment temperature of 4 °C has a synergistic effect on the inactivation of L. innocua, whereas post-treatment daylight illumination has an antagonic effect on PL antimicrobial efficacy. Post-PL-treatment temperature and illumination conditions could be thereby considered important environmental factors to activate, inhibit or control the repair and/or growth of L. innocua survivors after PL treatment., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Aims: The application of broad-spectrum intense light pulses is an innovative nonthermal technology for the decontamination of packaging materials, liquids or foodstuffs. The objective of this study was the fundamental investigation of the cellular impact of a pulsed light treatment on Listeria innocua and Escherichia coli., Methods and Results: Flow cytometry in combination with different fluorescent stains, conventional plate count technique and a viability assay were applied to investigate the effects of a pulsed light treatment on the physiological properties of L. innocua and E. coli. The results showed that loss of cultivability occurred at considerably lower fluences than the shutdown of cellular functions such as the depolarization of cell membranes, the loss of metabolic, esterase and pump activities or the occurrence of membrane damage. Therefore, a considerable proportion of cells appeared to have entered the viable but nonculturable (VBNC) state after the pulsed light treatment. A high percentage of L. innocua was able to maintain certain cellular vitality functions after storage overnight, whereas a further decrease in vitality was observed in case of E. coli. The loss of culturability was on the other hand directly accompanied by the formation of reactive oxygen species (ROS) and DNA damages, which were assessed by the ROS-sensitive probe DCFH-DA and RAPD-PCR, respectively., Conclusions: A significant discrepancy between conventional plate counts and different viability staining parameters was observed, which shows that a pulsed light treatment does not cause an immediate shutdown of vitality functions even when the number of colony-forming units already decreased for more than 6 log10 sample(-1) . Oxidative stress with concomitant damage to the DNA molecule showed to be directly responsible for the loss of cultivability due to pulsed light rather than a direct rupture of cell membranes or inactivation of intracellular enzymes., Significance and Impact of the Study: The presented results suggest an UV light-induced photochemical rather than a photothermal or photophysical inactivation of bacterial cells by pulsed light under the applied experimental conditions. Flow cytometry in combination with different viability stains proved to be a suitable technique to gain deeper insight into the cellular response of bacteria to inactivation processes like a pulsed light treatment., (© 2013 The Society for Applied Microbiology.)

The objective of this study was to produce phage display-derived binders with the ability to distinguish Listeria monocytogenes from other Listeria spp., which may have potential utility to enhance detection of Listeria monocytogenes. To obtain binders with the desired binding specificity a series of surface and solution phage-display biopannings were performed. Initially, three rounds of surface biopanning against gamma-irradiated L. monocytogenes serovar 4b cells were performed followed by an additional surface biopanning round against L. monocytogenes 4b which included prior subtraction biopanning against gamma-irradiated L. innocua cells. In an attempt to further enhance binder specificity for L. monocytogenes 4b two rounds of solution biopanning were performed, both rounds included initial subtraction solution biopanning against L. innocua. Subsequent evaluations were performed on the phage clones by phage binding ELISA. All phage clones tested from the second round of solution biopanning had higher specificity for L. monocytogenes 4b than for L. innocua and three other foodborne pathogens (Salmonella spp., Escherichia coli and Campylobacter jejuni). Further evaluation with five other Listeria spp. revealed that one phage clone in particular, expressing peptide GRIADLPPLKPN, was highly specific for L. monocytogenes with at least 43-fold more binding capability to L. monocytogenes 4b than to any other Listeria sp. This proof-of-principle study demonstrates how a combination of surface, solution and subtractive biopanning was used to maximise binder specificity. L. monocytogenes-specific binders were obtained which could have potential application in novel detection tests for L. monocytogenes, benefiting both the food and medical industries.

Pulsed light (PL) treatment can effectively inactivate a large proportion of contaminating bacteria on surfaces and in clear solutions. An important issue that needs to be investigated is whether repeated exposure to PL treatment causes any changes to the growth and resistance behavior of the bacteria surviving the treatment. To test this, three challenge microorganisms were used: Listeria monocytogenes, Listeria innocua, and Escherichia coli. Cells of the challenge bacteria were treated with either low or high PL doses. Survivors of the PL treatment were enumerated, isolated, regrown, and exposed again to PL treatment. PL inactivation curves were generated for the survivors of each exposure cycle (as well as controls) to examine possible differences induced by repeated treatments. Growth curves of L. monocytogenes, L. innocua, and E. coli isolates recovered from exposure to either 1.1 or 10.1 J/cm(2) were not significantly different from the growth curves of untreated cells. Reduction levels of up to 4 and up to 6 log CFU were obtained after exposure to 1.1 and 10.1 J/cm(2), respectively, both for the controls and the repeatedly treated and recovered isolates. These results show that PL did not significantly change the growth kinetics or resistance to PL of the target microorganisms after up to 10 exposures. These findings have significance for the practical application of PL treatment, as they indicate that this technology does not select for microorganisms with increased resistance.

The effect of β-lactoglobulin and β-casein on the pulsed light (PL) inactivation of Listeria innocua was evaluated. For low protein concentrations (β-lactoglobulin and β-casein up to 10 mg/mL), the lowest fluences applied (0.2 J/cm(2)) induced more than 7 Log reductions in cell counts. However, higher fluences were needed to induce similar reduction in L. innocua counts when this bacterium was suspended in solutions of higher protein concentration. The fluence required to induce similar microbial inactivation was lower in β-casein than in β-lactoglobulin solutions. For all protein solutions, the inactivation curves followed first order kinetics. The specific inactivation rate for L. innocua inactivation in protein solutions depended on the quantity of light transmitted in the range 230-290 nm by protein solutions. This work shows that PL technology could be used for the decontamination of high protein content solutions like whey or even higher protein concentration solutions., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Unlabelled: Recent outbreaks associated to the consumption of raw or minimally processed vegetable products that have resulted in several illnesses and a few deaths call for urgent actions aimed at improving the safety of those products. Electron beam irradiation can extend shelf-life and assure safety of fresh produce. However, undesirable effects on the organoleptic quality at doses required to achieve pathogen inactivation limit irradiation. Ways to increase pathogen radiation sensitivity could reduce the dose required for a certain level of microbial kill. The objective of this study was to evaluate the effectiveness of using natural antimicrobials when irradiating fresh produce. The minimum inhibitory concentration of 5 natural compounds and extracts (trans-cinnamaldehyde, eugenol, garlic extract, propolis extract, and lysozyme with ethylenediaminetetraacetate acid (disodium salt dihydrate) was determined against Salmonella spp. and Listeria spp. In order to mask odor and off-flavor inherent of several compounds, and to increase their solubility, complexes of these compounds and extracts with β-cyclodextrin were prepared by the freeze-drying method. All compounds showed bacteriostatic effect at different levels for both bacteria. The effectiveness of the microencapsulated compounds was tested by spraying them on the surface of baby spinach inoculated with Salmonella spp. The dose (D₁₀ value) required to reduce the bacterial population by 1 log was 0.190 kGy without antimicrobial addition. The increase in radiation sensitivity (up to 40%) varied with the antimicrobial compound. These results confirm that the combination of spraying microencapsulated antimicrobials with electron beam irradiation was effective in increasing the killing effect of irradiation., Practical Application: Foodborne illness outbreaks attributed to fresh produce consumption have increased and present new challenges to food safety. Current technologies (water washing or treating with 200 ppm chlorine) cannot eliminate internalized pathogens. Ionizing radiation is a viable alternative for eliminating pathogens; however, the dose required to inactivate these pathogens is often too high to be tolerated by the fresh produce without undesirable quality changes. This study uses natural antimicrobial ingredients as radiosensitizers. These ingredients were encapsulated and applied to fresh produce that was subsequently irradiated. The process results in high level of microorganism inactivation using lower doses than the conventional irradiation treatments., (© 2011 Institute of Food Technologists®)

High Intensity Light Pulses (HILP) represent an emerging processing technology which uses short (100-400 μs) light pulses (200-1100 nm) for product decontamination. In this study, model and real foods of differing transparencies (maximum recovery diluent (MRD), apple and orange juices and milk) were exposed to HILP in a batch system for 0, 2, 4 or 8 s at a frequency of 3 Hz. After treatment, inactivation of Escherichia coli or Listeria innocua was evaluated in pre-inoculated samples. Sensory and other quality attributes (colour, pH, Brix, titratable acidity, non-enzymatic browning, total phenols and antioxidant capacity (TEAC)) were assessed in apple juice. Microbial kill decreased with decreasing transparency of the medium. In apple juice (the most transparent beverage) E. coli decreased by 2.65 and 4.5 after exposure times of 2 or 4 s, respectively. No cell recovery was observed after 48 h storage at 4°C. No significant differences were observed in quality parameters, excepting TEAC and flavour score, where 8 s exposure caused a significant decrease (p<0.05). Based on these results, HILP with short exposure times could represent a potential alternative to thermal processing to eliminate undesirable microorganisms, while maintaining product quality, in transparent fruit juices., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The FDA recently approved irradiation treatment of leafy greens such as spinach up to 1 kGy; however, it is important to reduce the dose required to decontaminate the produce while maintaining its quality. Thus, the objectives of this study were: (1) to assess the radiation sensitivities of Salmonella spp. and Listeria spp. inoculated in ready-to-eat baby spinach leaves under modified atmosphere packaging (MAP) and irradiated using a 1.35-MeV Van de Graff accelerator (the leaves were irradiated both at room temperature and at -5 °C); and (2) to understand and optimize the synergistic effect of MAP and irradiation by studying the radiolysis of ozone formation under different temperatures, the effect of dose rate on its formation, and its decomposition. Results showed that increased concentrations of oxygen in the packaging significantly increased the radiation sensitivity of the test organisms, ranging from 7% up to 25% reduction in D(10)-values. In particular, radiosensitization could be effected (P < 0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Radiosensitization was demonstrated for both microorganisms with irradiation of either fresh or frozen (-5 °C) baby spinach. These results suggest that low-dose (below 1 kGy) e-beam radiation under modified atmosphere packaging (100% O(2) and N(2):O(2)[1:1]) may be a viable tool for reducing microbial populations or eliminating Salmonella spp. and Listeria spp. from baby spinach. A suggested treatment to achieve a 5-log reduction of the test organisms would be irradiation at room temperature under 100% O(2) atmosphere at a dose level of 0.7 kGy. Practical Application: Decontamination of minimally processed fruits and vegetables from food-borne pathogens presents technical and economical challenges to the produce industry. Internalized microorganisms cannot be eliminated by the current procedure (water-washed or treated with 200-ppm chlorine). The only technology available commercially is ionizing radiation; however, the actual radiation dose required to inactivate pathogens is too high to be tolerated by the product without unwanted changes. This study shows a new approach in using MAP with 100% O(2), which is converted to ozone to radiosensitize pathogens while improving the shelf life of minimally processed fruits and vegetables. The process results in a high level of microorganism inactivation using lower doses than the conventional irradiation treatments.

The efficiency of UV-C irradiation as a non-thermal pasteurization process for liquid egg white (LEW) was investigated. LEW inoculated with Escherichia coli K-12 (ATCC 25253), pathogenic strain of Escherichia coli O157:H7 (NCTC12900) and Listeria innocua (NRRL B33314) were treated with UV light using a bench top collimated beam apparatus. Inoculated LEW samples were exposed to UV-C irradiation of known UV intensity of 1.314mW/cm(2) and sample depth of 0.153cm for 0, 3 5, 7, 10, 13, 17 and 20min. The populations of E. coli K-12, E. coli O157:H7 and L. innocua were reduced after 20min of exposure by 0.896, 1.403 and 0.960logCFU respectively. Additionally, the inactivation data obtained for each strain suspended in LEW was correlated by using Weibull (2 parameter), Log-Linear (1 parameter), Hom (2 parameter) and modified Chick Watson (2 parameter) models. The inactivation kinetics of E. coli K-12, E. coli O157:H7 and L. innocua were best described by modified Chick Watson model with the smallest root mean squared error (RMSE) (R(2)> or =0.92)., (Copyright 2010 Elsevier B.V. All rights reserved.)

Listeria monocytogenes, a psychrotrophic foodborne pathogen, is a recurring postprocess contaminant on ready-to-eat meat (RTE) products including frankfurters. Flash (Steam) Pasteurization (FP) and ultraviolet light (254 nm-UVC) has been shown to reduce levels of L. monocytogenes and L. innocua on frankfurters. In this study, the use of UVC light followed by FP to inactivate L. innocua, a nonpathogenic surrogate for L.monocytogenes, on frankfurters that contained sodium diacetate and potassium lactate (SDA/PL) in a pilot-plant setting was investigated. Application of UVC (1.0 J/cm2), followed by FP (0.75 s steam/121 degrees C) resulted in inactivation of 3.19 log L. innocua, while application of UVC (4.0 J/cm2), followed by FP (3 s steam/121 degrees C) resulted in inactivation of 3.89 log of L. innocua. A refrigerated storage study (8 degrees C) of frankfurters that contained SDA/PL that were treated with UVC followed by FP revealed the growth of L. innocua was inhibited for approximately 8 wk following application of the interventions. The use of UVC in combination with FP had little effect on frankfurter color and texture. The combination of UVC, FP, and SDA/PL was found to be an effective hurdle process for decontamination of frankfurter surfaces.

The risk of listeriosis associated with ready-to-eat foods is a major concern in the United States. Pulsed light (PL) treatment has been effective for killing Listeria. The possibility of enhancing the antilisterial capability of PL treatment by combining PL with an additional hurdle, the natural antimicrobial nisin, was explored in this study. First, the ability of Listeria innocua to mimic the response of Listeria monocytogenes to PL treatment was demonstrated. Subsequently, a series of inoculation studies was performed in which canned sausages were surface inoculated with L. innocua as a surrogate for L. monocytogenes and then treated with a commercial preparation of nisin (Nisaplin), PL, or a combination of the two treatments. The application of a Nisaplin dip alone resulted in an immediate reduction of L. innocua by 2.35 +/- 0.09 log CFU. PL reduced L. innocua by 1.37 +/- 0.30 log CFU after exposure to 9.4 J/cm2. A total reduction of 4.03 +/- 0.15 log CFU was recorded after the combined treatment of Nisaplin and PL for 48 h at 4 degrees C. The long-term survival of L. innocua was evaluated on sausages stored at 4 degrees C. Treatment with Nisaplin and PL resulted in a 4- to 5-log reduction for two replicate studies. The combination treatment resulted in no significant microbial growth during 28 and 48 days of refrigerated storage in the first and second replicates, respectively. These results suggest that this combination treatment can be used as an effective antilisterial step in the production of ready-to-eat foods.

Optical density measurements were used to estimate the effect of heat treatments on the single-cell lag times of Listeria innocua fitted to a shifted gamma distribution. The single-cell lag time was subdivided into repair time (the shift of the distribution assumed to be uniform for all cells) and adjustment time (varying randomly from cell to cell). After heat treatments in which all of the cells recovered (sublethal), the repair time and the mean and the variance of the single-cell adjustment time increased with the severity of the treatment. When the heat treatments resulted in a loss of viability (lethal), the repair time of the survivors increased with the decimal reduction of the cell numbers independently of the temperature, while the mean and variance of the single-cell adjustment times remained the same irrespective of the heat treatment. Based on these observations and modeling of the effect of time and temperature of the heat treatment, we propose that the severity of a heat treatment can be characterized by the repair time of the cells whether the heat treatment is lethal or not, an extension of the F value concept for sublethal heat treatments. In addition, the repair time could be interpreted as the extent or degree of injury with a multiple-hit lethality model. Another implication of these results is that the distribution of the time for cells to reach unacceptable numbers in food is not affected by the time-temperature combination resulting in a given decimal reduction.

The inactivation kinetics for Listeria monocytogenes Scott A (CIP 103575, serotype 4b) and Listeria innocua (NTC 11288) after E-beam radiation were studied in vacuum-packed ready-to-eat dry-cured ham to optimize the sanitation treatment of this product. A treatment of 1.12 kGy was calculated to reach the food safety objective according to the U.S. Department of Agriculture criterion. No irradiation treatment is necessary to meet the European Union microbiological criterion for this bacterium. No changes (at doses < or =4 kGy) in the 2-thiobarbituric acid reactive substances values and texture were observed. Dry-cured hams treated with 1 and 2 kGy had negligible sensory modifications (appearance, odor, and flavor). However, the application of 3 and 4 kGy resulted in an increase in the intensity of off-odors and off-flavors. Despite these effects, all irradiated vacuum-packed dry-cured hams treated at < or =4 kGy were deemed acceptable for trading.

We investigated the degree of physiological damage to bacterial cells caused by optical trapping using a 1,064-nm laser. The physiological condition of the cells was determined by their ability to maintain a pH gradient across the cell wall; healthy cells are able to maintain a pH gradient over the cell wall, whereas compromised cells are less efficient, thus giving rise to a diminished pH gradient. The pH gradient was measured by fluorescence ratio imaging microscopy by incorporating a pH-sensitive fluorescent probe, green fluorescent protein or 5(6)-carboxyfluorescein diacetate succinimidyl ester, inside the bacterial cells. We used the gram-negative species Escherichia coli and three gram-positive species, Listeria monocytogenes, Listeria innocua, and Bacillus subtilis. All cells exhibited some degree of physiological damage, but optically trapped E. coli and L. innocua cells and a subpopulation of L. monocytogenes cells, all grown with shaking, showed only a small decrease in pH gradient across the cell wall when trapped by 6 mW of laser power for 60 min. However, another subpopulation of Listeria monocytogenes cells exhibited signs of physiological damage even while trapped at 6 mW, as did B. subtilis cells. Increasing the laser power to 18 mW caused the pH gradient of both Listeria and E. coli cells to decrease within minutes. Moreover, both species of Listeria exhibited more-pronounced physiological damage when grown without shaking than was seen in cells grown with shaking, and the degree of damage is therefore also dependent on the growth conditions.

Pulsed light (PL) treatment can effectively reduce microbial populations in clear substrates and on surfaces, but its effectiveness varies as a function of substrate or treatment-related factors. For PL to be successfully adopted by the food industry, all factors of influence, as well as the inactivation kinetics for the microorganisms of concern, must be elucidated. In this study, the inactivation kinetics of Listeria innocua and the effect of inoculum size on PL inactivation were investigated. Stainless steel coupons (50.8 by 101.6 mm) of defined surface properties and transparent glass chamber slides (25.4 by 50.8 by 10 mm) were each inoculated with 1 ml of aqueous suspensions of L. innocua containing inoculum populations of up to 10(9) CFU. The thickness of the liquid layer in the glass slides was 1.16 mm. The inoculated substrates were exposed to PL treatment of up to 17 J/cm2 in a static PL chamber equipped with a pulsed Xenon lamp. Survivors were recovered and enumerated by both standard plate counting and most-probable-number procedures. The data indicated that in clear liquids, PL resulted in more than a 6-log reduction of L. innocua after a 12-J/cm2 treatment, regardless of the initial inoculum size. For the stainless steel surfaces, less than a 4-log reduction after a 12-J/cm2 treatment and a noticeable effect of substrate characteristics and inoculum size on inactivation were observed. The survivor curves showed pronounced tailing for all substrates used in the study. The Weibull model accurately predicted the survivor ratios for the PL treatment of L. innocua in clear liquids, with a shape and scale parameter of 0.33 and 3.01, respectively. The Weibull model resulted in significant overestimation of PL effectiveness for the stainless steel substrates, where the influence of various substrate properties and inoculum level on inactivation was significant.

Biofilm formation on various surfaces is a well-known phenomenon and it has caused pollution, health, and safety hazards, and a substantial economic loss. The present study was to evaluate the bactericidal effects of sodium hypochlorite and gamma irradiation on Psudomonas aeruginosa, Listeria innocua, and Escherichia coli biofilm formed on polypropylene (PP), polyethylene (PET), and polycarbonate (PC), which are widely used as food container materials. The bacterial counts of all the micro-organisms tested in the bacterial suspension were decreased linearly by a gamma irradiation and 3 kGy of irradiation decreased the bacterial counts to below the detection limit (<10(1)cfu/ml). In sodium hypochlorite treated bacterial suspension only a 1 decimal point reduction in bacterial counts was observed until 100 ppm, beyond 100-400 ppm all micro-organisms tested were undetected. The microbial biofilms attached to PP, PE, and PC were very resistant to sodium hypochlorite, showing only 1-3 decimal point reductions even at 400 ppm of the total available chlorine level. In contrast, 3 kGy of gamma irradiation eliminated the micro-organisms attached to PP, PET, and PC with minor exceptions (P. aeruginosa attached to PE and Escherichia coli attached to PC). In conclusion, gamma irradiation was effective for reducing both the bacterial counts in the suspension and biofilms on PP, PET, and PC, while sodium hypochlorite was unable to eliminate the bacterial cells attached to PP, PET, and PC.

Pulsed light (PL) treatment is an alternative to traditional thermal treatment that has the potential to achieve several log-cycle reductions in the concentration of microorganisms. One issue that is still debated is related to what specifically causes cell death after PL treatments. The main objective of this work was to elucidate which portions of the PL range are responsible for bacterial inactivation. Stainless steel coupons with controlled surface properties were inoculated with a known concentration of Listeria innocua in the stationary growth phase and treated with 1 to 12 pulses of light at a pulse rate of 3 pulses per s and a pulse width of 360 micros. The effects of the full spectrum (lambda = 180 to 1,100 nm) were compared with the effects obtained when only certain regions of UV, visible, and near-infrared light were used. The effectiveness of the treatments was determined in parallel by the standard plate count and most-probable-number techniques. At a fluence of about 6 J/cm(2), the full-spectrum PL treatment resulted in a 4.08-log reduction of L. innocua on a Mill finish surface, the removal of lambda < 200 nm diminished the reduction to only 1.64 log, and total elimination of UV light resulted in no lethal effects on L. innocua. Overwhelmingly, the portions of the PL spectrum responsible for bacterial death are the UV-B and UV-C spectral ranges (X < 300 nm), with some death taking place during exposure to UV-A radiation (300 < lambda < 400 nm) and no observable death upon exposure to visible and near-infrared light (lambda > 400 nm). This work provides additional supporting evidence that cell death in PL treatment is due to exposure to UV light. Additionally, it was shown that even a minor modification of the light path or the UV light spectrum in PL treatments can have a significant negative impact on the treatment intensity and effectiveness.

Three ready-to-use vegetables, cucumber, blanched and seasoned spinach, and seasoned burdock were selected and the effects of an irradiation treatment for eliminating pathogens were investigated. The pathogens tested were Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus, and Listeria ivanovii. Inoculated viable cells of S. Typhimurium and L. ivanovii into cucumber and blanched and seasoned spinach were reduced about 4 decimal points by 2 kGy of irradiation and that of S. aureus inoculated into burdock showed about 4-decimal point reduction by 1 kGy. E. coli inoculated into burdock was not detected by 1 kGy. All the bacterial contents of test pathogens into the samples were reduced to below the limit of detection by 3 kGy irradiation. The range of the D10 value was 0.28-0.42 among the four pathogens. A Salmonella mutagenicity assay (Ames test) indicated that the 10 kGy-irradiated ready-to-use vegetables did not cause any increase. The studies indicated that a low-dose irradiation (3 kGy or less) can improve the microbial safety of ready-to-use vegetables.

The objective of this study was to evaluate the effect of an edible antimicrobial coating combined with modified atmosphere (MA) packaging (60% O2, 30% CO2, and 10% N2) and gamma irradiation on peeled minicarrots inoculated with Listeria innocua. Carrots were inoculated with L. innocua (10(3) CFU/g) and then coated with an antimicrobial coating based on calcium caseinate containing trans-cinnamaldehyde. The same formulation without trans-cinnamaldehyde was used as an inactive coating. Coated and uncoated carrots were packed under the MA or under air, irradiated at 0.25 or 0.5 kGy, and stored at 4 +/- 1 degrees C for 21 days. Samples were evaluated periodically for enumeration of L. innocua. Unirradiated carrots stored under air had the highest concentrations of L. innocua after 21 days of storage: 2.23 CFU/g in the uncoated samples and 2.26 CFU/ g in samples coated with the inactive coating. These results suggest that the inactive coating did not have any antimicrobial effect against L. innocua. However, the addition of the antimicrobial coating resulted in a 1.29-log reduction in the concentration of L. innocua in carrots packed under air after 21 days of storage and a 1.08-log reduction in carrots packed under MA after 7 days of storage. After 7 days of storage, no L. innocua was detected in samples treated at 0.5 kGy under air or in samples treated at 0.25 kGy under MA. A complete inhibition of L. innocua was also observed during all storage periods in uncoated and coated samples treated at 0.5 kGy under MA. These results indicate that the combination of irradiation and MA conditions play an important role in the radiosensitization of L. innocua.

Apple cider, inoculated with Escherichia coli and Listeria innocua, was processed using a simple UV apparatus. The apparatus consisted of a low-pressure mercury lamp surrounded by a coil of UV transparent tubing. Cider was pumped through the tubing at flow rates of 27 to 83 ml/min. The population of E. coli K-12 was reduced by 3.4 +/- 0.3 log after being exposed for 19 s at a treatment temperature of 25 degrees C. The population of L. innocua, which was more resistant to UV, was reduced by 2.5 +/- 0.1 log after being exposed for 58 s. The electrical energy for the process was 34 J/ml and is similar to that for conventional thermal processing. UV processing has the potential to improve the safety and extend the shelf life of apple cider.

Three prepared seafood products for manufacturing a laver (dried seaweed) roll, a traditional and rapidly growing ready-to-eat meal in Korea, were selected and the effects of irradiation treatment for eliminating pathogens of public health significance were investigated. The pathogens tested were Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus, and Listeria ivanovii. The radiation sensitivity (D10-values or the dose required to inactivate 90% of a population) of these organisms ranged from 0.23 to 0.62 kGy in imitation crab leg, 0.31 to 0.44 kGy in surimi gel, and 0.27 to 0.44 kGy in dried seaweed. The growth of all four test organisms inoculated (10(8) CFU/g) into these foods was inhibited by irradiation during 24 h of postirradiation storage regardless of the temperature (10, 20, and 30 degrees C). L. ivanovii was not detected after a 3-kGy treatment, but the other pathogens were not detected following irradiation at 2 kGy. These studies indicated that low-dose irradiation (2 kGy or less) of prepared seafood materials can keep them microbiologically safe before manufacturing a ready-to-eat prepared meal, a laver roll.

Ionizing radiation inactivates the pathogenic bacteria that can contaminate leafy green vegetables. Leaf pieces and leaf homogenate of endive (Cichorium endiva) were inoculated with the pathogen Listeria monocytogenes (ATCC 49594) or Listeria innocua (ATCC 51742), a nonpathogenic surrogate bacterium. The radiation sensitivity of the two strains was similar, although L. innocua was more sensitive to the type of suspending leaf preparation. During refrigerated storage after irradiation, the population of L. monocytogenes on inoculated endive was briefly suppressed by 0.42 kilogray (kGy), a dose calibrated to achieve a 99% reduction. However, the pathogen regrew after 5 days until it exceeded the bacterial levels on the control after 19 days in storage. Treatment with 0.84 kGy, equivalent to a 99.99% reduction, suppressed L. monocytogenes throughout refrigerated storage. Doses up to 1.0 kGy had no significant effect on the color of endive leaf material, regardless of whether taken from the leaf edge or the leaf midrib. The texture of leaf edge material was unaffected by doses up to 1.0 kGy, whereas the maximum dose tolerated by leaf midrib material was 0.8 kGy. These results show that endive leaves may be treated with doses sufficient to achieve at least a 99.99% reduction of L. monocytogenes with little or no impact on the product's texture or color.

Listeria spp. are a frequent postprocess contaminant of ready-to-eat (RTE) meat products, including ham. Vacuum-steam-vacuum (VSV) technology has been used successfully to eliminate Listeria innocua from hot dogs. Ionizing radiation can eliminate Listeria spp. from RTE meats. However, the excessive application of either technology can cause changes in product quality, including structural changes, changes in cure color (redness), and lipid oxidation. In this study, two cycles of VSV were combined with 2.0 kGy of ionizing radiation to obtain 4.40- and 4.85-log10 reductions of L. innocua on ham meat and skin, respectively. The use of both treatments resulted in an additive, as opposed to synergistic, reduction of L. innocua on ham. The combination treatment did not cause statistically significant changes in product structure, color (redness), or lipid oxidation.

Microorganisms in flowing water were disinfected by UV radiation from two excimer (excited dimer) lamps (emitting at 172 and 222 nm) in combination with two low-pressure mercury lamps (emitting at 254 nm). Synergies were investigated among the three types of radiation in the treatment of water spiked in turn with Escherichia coli, Listeria innocua, Shewanella putrefaciens, and spores of Bacillus subtilis and Bacillus cereus. Synergy was demonstrated between radiations at 222 and 254 nm in the treatment of E. coli, L. innocua, and S. putrefaciens, but little or no synergy was observed in the treatment of B. subtilis and B. cereus. At maximum flow rates (60 liters/min), 5-log reductions in E. coli were achieved at 254 nm, although at 222 nm, less than 1-log reductions were observed. No bacterial kill was observed with 172-nm radiation alone, despite increasing exposure time by reducing flow rates to less than 3 liters/min.

The effects of pulsed electric field (PEF) treatment and processing factors on the inactivation kinetics of Listeria innocua NCTC 11289 were investigated by using a pilot plant PEF unit with a flow rate of 200 liters/h. The electric field strength, pulse length, number of pulses, and inlet temperature were the most significant process factors influencing the inactivation kinetics. Product factors (pH and conductivity) also influenced the inactivation kinetics. In phosphate buffer at pH 4.0 and 0.5 S/m at 40 degrees C, a 3. 0-V/microm PEF treatment at an inlet temperature of 40 degrees C resulted in > or = 6.3 log inactivation of strain NCTC 11289 at 49.5 degrees C. A synergistic effect between temperature and PEF inactivation was also observed. The inactivation obtained with PEF was compared to the inactivation obtained with heat. We found that heat inactivation was less effective than PEF inactivation under similar time and temperature conditions. L. innocua cells which were incubated for a prolonged time in the stationary phase were more resistant to the PEF treatment, indicating that the physiological state of the microorganism plays a role in inactivation by PEF. Sublethal injury of cells was observed after PEF treatment, and the injury was more severe when the level of treatment was increased. Overall, our results indicate that it may be possible to use PEF in future applications in order to produce safe products.

Listeria spp. were sought in 32 chickens experimentally irradiated (2.5 kGy) and in 25 matched unirradiated chickens. Using the modified Food and Drug Administration method only three of the chickens were positive for L. monocytogenes. Using the more sensitive cold enrichment method 30 were found positive. The proportion of carcasses contaminated with L. monocytogenes was lower in the irradiated birds than in the unirradiated birds. The results also indicated that the numbers of L. monocytogenes in the irradiated birds were lower than in the unirradiated birds. The only other species of Listeria isolated was L. innocua, which was found on 44% of unirradiated chickens, but on none of the irradiated chickens.

The characteristics of the mononuclear phagocytes mediating resistance to infection with Listeria during the early phase (0 to 48 h) of the response have been investigated in genetically determined susceptible (A/J) and resistant (C57BL/6, B10.A/SgSn) strains of mice. Irradiation immediately before infection profoundly enhanced the bacterial growth in the resistant strain, while having no effect in the susceptible strain, over a wide range (3 x 10(3) to 10(5)) of infective doses. This effect of irradiation is demonstrable at low-dose radiation (200 roentgens) and can be reversed by repopulation with 20 x 10(6) syngeneic nucleated bone marrow cells. Administration of dextran sulfate 500 24 h before infection profoundly enhanced the bacterial growth in the susceptible strain, while having much less effect in the resistant strain. Thus, the genetic advantage of the resistant mouse strains to listerial infection, at least during the early phase of the response, appears to be due to a cellular mechanism that is highly radiosensitive and relatively insensitive to dextran sulfate 500. In the susceptible strain, the early protective cellular mechanism is radioresistant and highly dextran sulfate 500 sensitive.