Antibacterial action of synthetic antilipopolysaccharide peptides (SALP) involves neutralization of both membrane-bound and free toxins.

Increasing failure of conventional antibiotics to combat bacterial infections requires the urgent development of new antibacterial drugs; a promising class of new drugs based on antimicrobial peptides. Here, we studied the molecular interaction of polycationic synthetic antilipopolysaccharide peptides (SALPs) with various gram-negative and gram-positive bacteria, including resistant strains. The analysis of antimicrobial activity by conventional techniques and atomic force microscopy showed a strict dependence on amino acid (aa) sequences, with the type of amino acid, its position within the primary structure, and the sequence length being critical parameters. By monitoring lipopolysaccharide (LPS)- or bacteria-induced cytokine production in human mononuclear cells and whole blood, we found a direct link between the binding of the lead compound Pep19-2.5 to Salmonella enterica and the anti-inflammatory activity of the peptide. Thermodynamic analysis of Pep19-2.5 binding to the bacterial cell envelope showed an exothermic reaction with saturation characteristics, whereas small-angle X-ray scattering data indicated a direct attachment of Pep19-2.5 to the bacterial cell envelope. This binding preferentially takes place to the LPS outer monolayer, as evidenced by the change in the LPS acyl chain and phosphate vibrational bands seen by Fourier-transform infrared spectroscopy. We report here that the anti-inflammatory activity of Pep19-2.5 is not only connected with neutralization of cell-free bacterial toxins but also with a direct binding of the peptide to the outer leaflet of the bacterial outer membrane.

Salt content dependent dielectric properties of pistachios relevant to radio-frequency pasteurization.

This study was conducted to investigate the effect of salt content during radio-frequency (RF) heating on rate of temperature increase, dielectric properties (DPs), and reduction of pathogens in pistachios. Also, the effect of RF heating on pistachio quality of varying salt content was determined. Pistachios of different salt content (0, 100, and 330 mg sodium/serving) were inoculated with Salmonella enterica and treated in a 27.12-MHz RF heater. The RF heating rate increased when salt content was in the range of 0-100 mg sodium/serving, but there were no significant (P > 0.05) differences in the rate of temperature rise after salt content reached to 100 mg sodium/serving. Both dielectric constant and dielectric loss factor of pistachios increased with rising salt content. Along with increased salt content, RF treatment time required to reduce this pathogen by 4 log CFU/g decreased first and then remained the same above an upper limit of salt content corresponding to the peak heating rate. RF treatment did not significantly (P > 0.05) cause changes in the color and level of lipid oxidation of pistachios. The results of the current study imply that RF heating may be a potential intervention for inactivating foodborne pathogens in pistachios and that the effect of pasteurization is influenced by dielectric loss factor relative to salt content.

Sanitation of tomatoes based on a combined approach of washing process and pulsed light in conjunction with selected disinfectants.

In this study, we evaluated the performance of a large-scale decontamination system based on a washing process in combination with pulsed light (PL) exposure and H2O2/chlorine. In order to identify optimum processing condition, we first evaluated the effect of single and combined PL treatments on the inactivation of Salmonella on grape tomatoes using a small sample size of 50 g. Two inoculation methods, spot and dip, were used to simulate different contamination scenarios and two wash water quality, clear tap water and turbid tap water with extremely high levels of organic load and soil, were used to represent clean and very dirty wash water. In general, the combined PL-Chlorine and PL-H2O2 treatments were more or as effective as chlorine washing in killing Salmonella on grape tomatoes and were able to keep residual Salmonella in wash water below the detection limit of 2 CFU/mL. The PL alone and combined PL-H2O2 treatments were chosen and further tested for their decontamination efficacy under turbid wash water condition using large sample sizes, 300, 1000 and 2000 g. Sample size did not negatively affect the single and combined PL treatments on the inactivation of Salmonella on grape tomatoes. The combined PL-H2O2 treatment in general showed better inactivation effect of Salmonella on tomatoes than the PL alone treatment. Additionally, the combined PL-H2O2 treatment reduced Salmonella in turbid wash water below the detection limit of 2 CFU/mL in the majority of cases. In conclusion, the combined PL-H2O2 treatment could potentially be used as an environmentally friendly alternative to chlorine washing for tomato decontamination and cleaning.

Individual and combined efficacies of mild heat and ultraviolet-c radiation against Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in coconut liquid endosperm.

This study determined the inactivation kinetic parameters of selected pathogens in heat, ultraviolet-C and combined heat-UV-C treated coconut liquid endosperm. Separate cocktails of Escherichia coli O157:H7, Salmonella enterica serovars, and Listeria monocytogenes strains were inoculated into coconut liquid endosperm (pH 5.15, TSS 4.4oBx, TA 0.062% malic acid, extinction coefficient (ε) at 254 nm of 0.0154 cm-1) for inactivation studies. Result showed that all organisms generally exhibited a log-linear heat inactivation behavior (R2 0.81-0.99). The E. coli O157:H7 cocktail (D55 = 19.75 min, D57 = 10.79 min, D60 = 3.38 min, and D63 = 0.46 min) was found to be significantly more resistant (P > 0.05) than the tested cocktail of L. monocytogenes (D55 = 11.68 min, D57 = 4.53 min, D60 = 1.82 min and D63 = 0.26 min) and S. enterica cocktail (D55 = 3.08 min, D57 = 2.60 min, D60 = 0.89 min and D63 = 0.25 min). Despite the differences in DT values, computed z values for L. monocytogenes cocktail (5.12 ±â€¯0.43 °C) and E. coli O157:H7 cocktail (4.95 ±â€¯0.12 °C) were not significantly different (P > 0.05), but were both significantly (P < 0.05) lower than that of S. enterica cocktail (7.10 ±â€¯0.15 °C). All test organisms also exhibited a generally log-linear UV-C inactivation behavior (R2 0.90-0.99) with E. coli O157:H7 cocktail (DUV-C = 25.26 mJ/cm2) demonstrating greatest resistance to UV-C than S. enterica (DUV-C = 24.65 mJ/cm2) and L. monocytogenes (DUV-C = 17.30 mJ/cm2) cocktails. The D55 values of each organism cocktail were used to calculate for the 3-log reduction heating process schedules, during which UV-C treatments were simultaneously applied. Lethal rates (F values) calculations in the combined processes revealed that within the 3-log reduction heating processes, co-exposure of UV-C resulted in 5.62 to 6.20 log reductions in the test organism populations. Heating caused 69.3, 97.2, and 67.4% of the reduction in E. coli O157:H7, S. enterica and L. monocytogenes cocktails, respectively. These results can be used as baseline data in the establishment of mild heat treatment in combination with UV-C process schedules for coconut liquid endosperm and other similar products.

A model for the influences of soluble and insoluble solids, and treated volume on the ultraviolet-C resistance of heat-stressed Salmonella enterica in simulated fruit juices.

This study was conducted to determine the effects of intrinsic juice characteristics namely insoluble solids (IS, 0-3 %w/v), and soluble solids (SS, 0-70 °Brix), and extrinsic process parameter treated volume (250-1000 mL) on the UV-C inactivation rates of heat-stressed Salmonella enterica in simulated fruit juices (SFJs). A Rotatable Central Composite Design of Experiment (CCRD) was used to determine combinations of the test variables, while Response Surface Methodology (RSM) was used to characterize and quantify the influences of the test variables on microbial inactivation. The heat-stressed cells exhibited log-linear UV-C inactivation behavior (R2 0.952 to 0.999) in all CCRD combinations with DUV-C values ranging from 10.0 to 80.2 mJ/cm2. The DUV-C values obtained from the CCRD significantly fitted into a quadratic model (P < 0.0001). RSM results showed that individual linear (IS, SS, volume), individual quadratic (IS2 and volume2), and factor interactions (IS × volume and SS × volume) were found to significantly influence UV-C inactivation. Validation of the model in SFJs with combinations not included in the CCRD showed that the predictions were within acceptable error margins.

Inactivation of Gram (-) bacteria Salmonella enterica by chlorophyllin-based photosensitization: Mechanism of action and new strategies to enhance the inactivation efficiency.

This study is focused on the enhancement of susceptibility of Gram (-) bacteria S. enterica to chlorophyllin-based (Chl) photosensitization combining it with other antimicrobial tools. In order to find best combinations, the mechanism by which Chl-based photosensitization inactivates bacteria must be identified. Data confirmed that photosensitization (Chl 1.5×10-5M, for 1-120min, 405nm, 0-46.1J/cm2) reduced S. enterica population, just by 2.05 log (CFU/ml). Fluorimetric measurements indicated that just minor part of Chl was bound to Salmonella in suspension. Addition of sodium azide (NaN3) (10mM) protected bacteria from killing, what means that 1O2 took place in photochemical reactions. Gene expression data confirmed that Chl-based photosensitization induced oxidative stress in bacteria cells, since mostly genes responsible for detoxification of ROS (OxyR, AhpC, GrxA) have been expressed in Salmonella. Moreover, the expression of genes, responsible for the inhibition of oxidative respiration (AtpC), cell division and down-regulation of metabolism (SulA) have been detected. In addition, Chl-based photosensitization induced significant release of intracellular components (absorbing at λ260 nm and λ280 nm) in bacteria that indicated increased membrane permeability. Thus, the combination of two antimicrobials (Chl-based photosensitization and chitosan (CHS)) with the same target (cellular membrane) in the presence of light drastically reduced viable Salmonella population (by 7.28 log). Combined treatment of photosensitization and high power pulsed UV light (HPPL) was also very effective, since reduced viable Salmonella by 7.5 log. Bacterial regrowth experiments clearly indicated that after both combined treatments Salmonella lost its ability to proliferate, and SEM images confirmed that after both treatments no viable bacteria have been found at all.

Influence of static magnetic field exposure on fatty acid composition in Salmonella Hadar.

We have been interested, in this work, to investigate the effect of the exposure to static magnetic field at 200 mT (SMF) on the fatty acid (FA) composition of Salmonella enterica subsp Enterica serovar Hadar isolate 287: effects on the proportion of saturated and unsaturated fatty acids (SFAs, UFAs), cyclopropane fatty acids (CFAs) and hydroxy fatty acids after exposure to the static magnetic field at 200 mT (SMF). Analysis with Gas Chromatography-Mass Spectrometry (GC-MS) of total lipid showed that the proportion of the most fatty acids was clearly affected. The comparison of UFAs/SFAs ratio in exposed bacteria and controls showed a diminution after 3 and 6 h of exposure. This ration reached a balance after 9 h of treatment with SMF. So we can conclude that S. Hadar tries to adapt to magnetic stress by changing the proportions of SFAs and UFAs over time to maintain an equilibrium after 9 h of exposure, thus to maintain the inner membranes fluidity. Also, a decrease in the proportion of hydroxy FAs was observed after 6 h but an increase of this proportion after 9 h of exposure. Concerning CFAs, its proportion raised after 6 h of exposure to the SMF but it decreased after 9 h of exposure. These results are strongly correlated with those of cfa (cyclopropane fatty acid synthase) gene expression which showed a decrease of its expression after 9 h of exposure.

Impact of pulsed light on cellular activity of Salmonella enterica.

AIMS: The objective of this study was a comprehensive characterization of physiological changes of Salmonella enterica induced by intense broad spectrum pulsed light (PL). After exposing the bacteria to this nonthermal decontamination technology on a gel surface, multiple viability parameters beyond culturability were assessed. METHODS AND RESULTS: By applying flow cytometry, a luciferin-luciferase bioluminescence assay and a microplate assay to measure the current redox activity, the impact of pulsed light on the membrane potential, membrane integrity, esterase activity, efflux pump activity, expression of the green fluorescent protein (GFP), respiration activity and ATP-content of Salm. enterica ATCC BAA-1045 was determined. These culture-independent methods for assessing the bacterial activity were compared to the ability to grow on tryptic soy agar. It is shown that this strain is rather sensitive to PL considering colony count reductions, while on the other hand unculturable bacteria still exhibit significant cellular energetic functions. However, this residual activity after PL exposure significantly decreases during sample storage in buffer for 24 h. This study also shows that the GFP expression of PL-treated cells which have rendered unculturable is severely reduced. CONCLUSIONS: This study reveals that although not all cellular functions of Salm. enterica are immediately shut down after PL exposure, the synthesis of new GFP is strongly reduced and affected to a similar extent as the culturability. SIGNIFICANCE AND IMPACT OF THE STUDY: It is shown for the first time, that even there is significant bacterial activity measurable after PL exposure, it is likely that nongrowing pathogenic bacteria like Salm. enterica are unable to express proteins, which is of great importance regarding their pathogenicity.

Effective photosensitization-based inactivation of Gram (-) food pathogens and molds using the chlorophyllin-chitosan complex: towards photoactive edible coatings to preserve strawberries.

This study is focused on the novel approaches to enhance the inactivation of the Gram (-) food pathogen Salmonella enterica and harmful molds in vitro and on the surface of strawberries using the chlorophyllin-chitosan complex. Salmonella enterica (∼1 × 10(7) CFU mL(-1)) was incubated with chlorophyllin 1.5 × 10(-5) M (Chl, food additive), chitosan 0.1% (CHS, food supplement) or the chlorophyllin-chitosan complex (1.5 × 10(-5) M Chl-0.1% CHS) and illuminated with visible light (λ = 405 nm, light dose 38 J cm(-2)) in vitro. Chlorophyllin (Chl)-based photosensitization inactivated Salmonella just by 1.8 log. Chitosan (CHS) alone incubated for 2 h with Salmonella reduced viability 2.15 log, whereas photoactivated Chl-CHS diminished bacterial viability by 7 log. SEM images indicate that the Chl-CHS complex under these experimental conditions covered the entire bacterial surface. Significant cell membrane disintegration was the main lethal injury induced in Gram (-) bacteria by this treatment. Analysis of strawberry decontamination from surface-inoculated Salmonella indicated that photoactivated Chl-CHS (1.5 × 10(-5) M Chl-0.1% CHS, 30 min incubation, light dose 38 J cm(-2)) coatings diminished the pathogen population on the surface of strawberries by 2.2 log. Decontamination of strawberries from naturally distributed yeasts/molds revealed that chitosan alone reduced the population of yeasts/molds just by 0.4 log, Chl-based photosensitization just by 0.9 log, whereas photoactivated Chl-CHS coatings reduced yeasts/molds on the surface of strawberries by 1.4 log. Electron paramagnetic resonance spectroscopy confirmed that no additional photosensitization-induced free radicals have been found in the strawberry matrix. Visual quality (color, texture) of the treated strawberries was not affected either. In conclusion, photoactive Chl-CHS exhibited strong antimicrobial action against more resistant to photosensitization Gram (-) Salmonella enterica in comparison with Gram (+) bacteria in vitro. It reduced significantly the viability of strawberry surface-attached yeasts/molds and inoculated Salmonella without any negative impact on the visual quality of berries. Experimental data support the idea that photoactivated Chl-CHS can be a useful tool for the future development of edible photoactive antimicrobial coatings which can preserve strawberries and prolong their shelf-life according to requirements of "clean green technology".

Effect of X-ray treatments on Escherichia coli O157:H7, Listeria monocytogenes, Shigella flexneri, Salmonella enterica and inherent microbiota on whole mangoes.

UNLABELLED: The aims of this investigation were to; (i) study the effect of X-ray treatments in reducing Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on whole mangoes, and (ii) study the effect of X-ray treatments on microflora counts (mesophilic counts, psychrotrophic counts and yeast and mould counts) of whole mangoes during storage at ambient temperature (22°C) for 30 days. A mixture of three strains of each test organism was spot inoculated (100 µl; approx. 8-9 log CFU ml(-1) ), separately, onto the surface (5 cm(2) ) of whole mangoes, air-dried (30 min), and then treated with different doses of X-ray (0, 0·1, 0·5, 1·0, and 1·5 kGy). Approximately 2·9, 1·8, 2·1 and 5·2 log CFU cm(-2) reduction of E. coli O157:H7, L. monocytogenes, Sh. flexneri and Salm. enterica were achieved by treatment with 0·5 kGy X-ray respectively. Furthermore, the populations of E. coli O157:H7, L. monocytogenes, Sh. flexneri and Salm. enterica were reduced to less than the detectable limit (2·0 log CFU cm(-2) ) by treatment with 1·5 kGy X-ray. Treatment with 1·5 kGy X-ray significantly reduced the initial inherent microflora on skin of whole mangoes and inherent levels were significantly (P < 0·05) lower than the control sample throughout storage at 22°C for 30 days. SIGNIFICANCE AND IMPACT OF THE STUDY: Fresh produce was associated with 770 outbreaks between 1990 and 2005, resulting in 35 060 cases of illness that costs the US $39 billion annually. The food industry is looking for new preservation methods. This investigation indicated that X-ray treatment was very effective against Escherichia coli O157:H7, Listeria monocytogenes, Shigella flexneri and Salmonella enterica and inherent microflora on whole mangoes which could offer an applicable approach to control pathogens and spoilage bacteria for the mango industry.

Inactivation of Escherichia Coli O157:H7 and Salmonella Enterica on Blueberries in Water Using Ultraviolet Light.

UNLABELLED: Ultraviolet light (UV) has antimicrobial effects, but the shadowing effect has limited its application. In this study, a novel setup using UV processing in agitated water was developed to inactivate Escherichia coli O157:H7 and Salmonella on blueberries. Blueberries were dip- or spot-inoculated with E. coli or Salmonella. Blueberries inoculated with E. coli were treated for 2 to 10 min with UV directly (dry UV) or immersed in agitated water during UV treatment (wet UV). E. coli was most easily killed on spot-inoculated blueberries with a 5.2-log reduction after 10-min wet UV treatment. Dip-inoculated blueberries were the most difficult to be decontaminated with only 1.6-log reduction after 10-min wet UV treatment. Wet UV treatment generally showed higher efficacies than dry UV treatment, achieving an average of 1.4 log more reduction for spot-inoculated blueberries. For dip-inoculated blueberries, chlorine washing and UV treatments were less effective, achieving <2 log reductions of E. coli. Thus, the efficacy of combinations of wet UV with sodium dodecyl sulfate (SDS), levulinic acid, or chlorine was evaluated. Inoculated blueberries were UV-treated while being immersed in agitated water containing 100 ppm SDS, 0.5% levulinic acid or 10 ppm chlorine. The 3 chemicals did not significantly enhance the wet UV treatment. Findings of this study suggest that UV treatment could be used as an alternative to chlorine washing for blueberries and potentially for other fresh produce. PRACTICAL APPLICATION: A novel UV light system for decontamination of blueberries in water was developed and evaluated. Results demonstrated that the decontamination efficacy of this system was generally as effective as chlorine washing, indicating that it could potentially be used as an alternative to chlorine washing for blueberries and other fresh produce.

Reduction of Salmonella enterica serotype Poona and background microbiota on fresh-cut cantaloupe by electron beam irradiation.

The efficacy of electron beam (e-beam) irradiation processing to reduce Salmonella enterica serotype Poona on surfaces of fresh-cut cantaloupe, and the impact of e-beam irradiation processing on the numbers of indigenous microorganisms were determined. Additionally, the D10-value for S. Poona reduction on the cut cantaloupe was also determined. Fresh-cut cantaloupe pieces, inoculated with S. Poona to 7.8 log10 CFU/g, were exposed to 0.0, 0.7, or 1.5 kGy. Surviving S. Poona, lactic acid bacteria (LAB), and fungi (yeasts, molds) were periodically enumerated on appropriate media over 21 days of storage at 5 °C. Cantaloupe surface pH was measured for irradiated cantaloupe across the 21 day storage period. To determine the D10-value of S. Poona, cantaloupe discs were inoculated and exposed to increasing radiation dosages between 0 and 1.06 kGy; surviving pathogen cells were selectively enumerated. S. Poona was significantly reduced by irradiation; immediate reductions following exposure to 0.7 and 1.5 kGy were 1.1 and 3.6 log10 CFU/g, respectively. After 21 days, S. Poona numbers were between 4.0 and 5.0 log10 CFU/g less than untreated samples at zero-time. Yeasts were not reduced significantly (p ≥ 0.05) by e-beam irradiation and grew slowly but steadily during storage. Counts of LAB and molds were initially reduced with 1.5 kGy (p<0.05) but then LAB recovered grew to high numbers, whereas molds slowly declined for irradiated and control samples. Cantaloupe pH declined during storage, with the greatest decrease in untreated control cantaloupe (p<0.05). The D10-value for S. Poona was determined to be 0.211 kGy, and this difference from the reductions observed in the cut cantaloupe studies may be due to the more precise dose distribution obtained in the thin and flat cantaloupe pieces used for the D10-value experiments. The effect of e-beam irradiation at the same doses used in this study was determined in previous studies to have no negative effect in the quality of the cut cantaloupe. Therefore, incorporation of low dosage ionizing irradiation and consistent application of irradiation processing can significantly improve the microbiological safety of fresh-cut cantaloupe.

Previous physicochemical stress exposures influence subsequent resistance of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes to ultraviolet-C in coconut liquid endosperm beverage.

This study investigated the influences of prior exposures to common physicochemical stresses encountered by microorganisms in food and food processing ecologies such as acidity, desiccation, and their combinations, on their subsequent susceptibility towards UV-C treatment in coconut liquid endosperm beverage. Cocktails of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes were separately subjected to gradually acidifying environment (final pH 4.46), exposed to abrupt desiccation by suspension in saturated NaCl solution (aw=0.85) for 4, 8, and 24h, and sequential acidic and desiccated stresses before suspending in the coconut beverage for UV-C challenge. The exposure times (D) and UV-C energy dose values (DUV-C) necessary to reduce 90% of the population of the different test organisms varied with previous exposures to different sublethal stresses, indicating possible influence of implicit microbial factors towards resistance to UV-C. All tested individual and combined stresses resulted in increased resistance, albeit some were not statistically significant. Non-stressed cells had D values of 3.2-3.5s, and corresponding DUV-C values of 8.4-9.1 mJ/cm(2). Cells exposed to previous acid stress had D values of 4.1-4.8s and corresponding DUV-C values of 10.7-12.5 mJ/cm(2). Prior exposure to desiccation resulted in D values of 5.6-7.9s and DUV-C values of 14.7-20.6 mJ/cm(2), while exposure to combined acid and desiccation stresses resulted in D values of 6.1-8.1s and DUV-C values of 15.9-21.0 mJ/cm(2). The D and DUV-C values of S. enterica after previous exposure to sequential acid (24 h) and desiccation (24 h) stresses were found significantly greatest, making the organism and physiological state an appropriate reference organism for the establishment of UV-C pasteurization process for the beverage.

Feverlike Temperature is a Virulence Regulatory Cue Controlling the Motility and Host Cell Entry of Typhoidal Salmonella.

Human infection with typhoidal Salmonella serovars causes a febrile systemic disease, termed enteric fever. Here we establish that in response to a temperature equivalent to fever (39 °C-42 °C) Salmonella enterica serovars Typhi, Paratyphi A, and Sendai significantly attenuate their motility, epithelial cell invasion, and uptake by macrophages. Under these feverlike conditions, the residual epithelial cell invasion of S. Paratyphi A occurs in a type III secretion system (T3SS) 1-independent manner and results in restrained disruption of epithelium integrity. The impaired motility and invasion are associated with down-regulation of T3SS-1 genes and class II and III (but not I) of the flagella-chemotaxis regulon. In contrast, we demonstrate up-regulation of particular Salmonella pathogenicity island 2 genes (especially spiC) and increased intraepithelial growth in a T3SS-2-dependent manner. These results indicate that elevated physiological temperature is a novel cue controlling virulence phenotypes in typhoidal serovars, which is likely to play a role in the distinct clinical manifestations elicited by typhoidal and nontyphoidal salmonellae.

UV-C pre-adaptation of Salmonella: effect on cell morphology and membrane fatty acids composition.

The present study was carried out to evaluate the effects of ultraviolet radiations (UV-C) on the fatty acids composition of three serovars of Salmonella: S. typhimurium, S. hadar and S. zanzibar. Results obtained show that UV-C treatment increases significantly (P ≤ 0.05) the percentage of cyclic fatty acids. The atomic force microscopy was used to study the morphology and cell surface of irradiated strains. Results show that UV-C rays induce morphological changes and alter the bacterial cell surface (presence of grooves and irregularities).

Increased water activity reduces the thermal resistance of Salmonella enterica in peanut butter.

Increased water activity in peanut butter significantly (P < 0.05) reduced the heat resistance of desiccation-stressed Salmonella enterica serotypes treated at 90 °C. The difference in thermal resistance was less notable when strains were treated at 126 °C. Using scanning electron microscopy, we observed minor morphological changes of S. enterica cells resulting from desiccation and rehydration processes in peanut oil.

Efficacy of integrated treatment of UV light and low-dose gamma irradiation on inactivation of Escherichia coli O157:H7 and Salmonella enterica on grape tomatoes.

The study evaluated the efficacy of integrated ultraviolet-C light (UVC) and low-dose gamma irradiation treatments to inactivate mixed strains of Escherichia coli O157:H7 and Salmonella enterica on inoculated whole grape tomatoes. A mixed bacterial cocktail composed of a 3 strain mixture of E. coli O157:H7 (C9490, E02128, and F00475) and a 3 serotype mixture of S. enterica (S. Montevideo G4639, S. Newport H1275, and S. Stanley H0558) was used based on their association with produce-related outbreaks. Spot inoculation (50 to 100 µmL) on tomato surfaces was performed to achieve a population of appropriately 10(7-8) CFU/tomato. Inoculated tomatoes were subjected to UVC (253.7 nm) dose of 0.6 kJ/m(2) followed by 4 different low doses of gamma irradiations (0.1 kGy, 0.25 kGy, 0.5 kGy, 0.75 kGy). The fate of background microflora (mesophilic aerobic) including mold and yeast counts were also determined during storage at 5 °C over 21 d. Integrated treatment significantly (P < 0.05) reduced the population of target pathogens. Results indicate about 3.4 ± 0.3 and 3.0 ± 0.1 log CFU reduction of E. coli O157:H7 and S. enterica, respectively, per tomato with UVC (0.6 kJ/m(2) ) and 0.25 kGy irradiation. More than a 4 log and higher reduction (>5 log) per fruit was accomplished by combined UVC treatment with 0.5 kGy and 0.75 kGy irradiation, respectively, for all tested pathogens. Furthermore, the combined treatment significantly (P < 0.05) reduced the native microflora compared to the control during storage. The data suggest efficacious treatment strategy for produce indicating 5 or higher log reduction which is consistent with the recommendations of the Natl. Advisory Committee on Microbiological Criteria for Foods.

Solar inactivation of four Salmonella serovars in fresh and marine waters.

Sunlight-mediated disinfection of water is of interest to both the drinking and recreational water quality community of researchers due to its potential to reduce microbial contamination and waterborne illness. Photo-inactivation of enteric bacteria has primarily been investigated using Escherichia coli and laboratory strains of model bacteria. The present study sought to document the photo-inactivation of environmental isolates of Salmonella in filter-sterilized natural seawater and freshwater and to test the hypothesis that diverse Salmonella serovars decay at similar rates both within and between water matrices. The inactivation of Salmonella enterica Typhimurium LT2, Typhimurium ST19, Heidelberg, and Mbandaka was examined in sunlit and dark microcosms. First order decay was observed in sunlit microcosms; the time until 90% inactivation was of the order of 10 min. A significant shoulder, of the order of 1 hr in length, was observed in the freshwater microcosms during which concentrations were stable. Serovar Mdandaka decayed more slowly than other serovars in both seawater and freshwater. The serovars were extremely stable in the dark microcosms showing little to no decay over 53 days. The results document intra-species variation in photo-inactivation, likely owing to differences in intracellular concentrations of photo-sensitizing molecules or molecules that quench reactive species.

Effect of x-ray treatments on pathogenic bacteria, inherent microbiota, color, and texture on parsley leaves.

This work is a part of systematic studies of the effect of X-ray treatments on fresh produce. The main objective of this investigation was to study the effects of X-ray treatments in reducing the concentration of artificially inoculated Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica, and Shigella flexneri, and inherent microbiota on parsley leaves. The secondary objective was to study the effects of X-ray treatments on color and texture parameters on treated parsley leaves. The Dip-inoculated method was used to inoculate parsley leaves with a mixture of two or three strains of each tested organism at 10(8) to 10(9) colony-forming unit (CFU)/mL; the inoculated parsley leaves were then air-dried and followed by treatment with different doses of X-ray (0, 0.1, 0.5, 1.0, and 1.5 kGy) at 22°C and 55-60% relative humidity. Surviving bacterial populations on parsley leaves were evaluated using a nonselective medium (tryptic soy agar) with a selective medium overlay for each bacterium: E. coli O157:H7 (CT-SMAC agar), L. monocytogenes (MOA), and S. enterica and S. flexneri (XLD). Approximately 5.8, 3.1, 5.7, and 5.2 log CFU reductions of E. coli O157:H7, L. monocytogenes, S. enterica, and Shigella flexneri were achieved by treatment with 1.0 kGy X-ray, respectively. Furthermore, the populations of E. coli O157:H7, L. monocytogenes, S. enterica, and Shigella flexneri were reduced to less than the detectable limit (1.0 log CFU/g) by treatment with 1.5 kGy X-ray. Treatment with 1.5 kGy X-ray significantly reduced the initial inherent microbiota on parsley leaves, and inherent levels were significantly (p < 0.05) lower than the control sample throughout refrigerated storage for 30 days. No significant differences (p > 0.05) in color or texture of control and treated samples with 0.1-1.5 X-ray were observed. The results of investigation indicated that X-ray is an effective technology to eliminate E. coli O157:H7, L. monocytogenes, S. enterica, and Shigella flexneri, and to extend the shelf life of parsley leaves.

Bactericidal effects of 405 nm light exposure demonstrated by inactivation of Escherichia, Salmonella, Shigella, Listeria, and Mycobacterium species in liquid suspensions and on exposed surfaces.

The bactericidal effect of 405 nm light was investigated on taxonomically diverse bacterial pathogens from the genera Salmonella, Shigella, Escherichia, Listeria, and Mycobacterium. High-intensity 405 nm light, generated from an array of 405-nm light-emitting diodes (LEDs), was used to inactivate bacteria in liquid suspension and on exposed surfaces. L. monocytogenes was most readily inactivated in suspension, whereas S. enterica was most resistant. In surface exposure tests, L. monocytogenes was more susceptible than Gram-negative enteric bacteria to 405 nm light when exposed on an agar surface but interestingly less susceptible than S. enterica after drying onto PVC and acrylic surfaces. The study findings, that 405 nm light inactivates diverse types of bacteria in liquids and on surfaces, in addition to the safety advantages of this visible (non-UV wavelength) light, indicate the potential of this technology for a range of decontamination applications.