Antimicrobial performance of alkaline ionic fluid (GC-100X) and its ability to remove Escherichia coli O157:H7 from the surface of tomatoes

An efficacy test of GC-100X, a noncorrosive alkaline ionic fluid (pH 12) composed of free radicals and supplemented with xylitol, was carried out against six major foodborne pathogens-Staphylococcus aureus FRI 913, Salmonella enterica serovar Enteritidis ATCC 13076, S. enterica serovar Typhimurium DT104 Korean isolate, Vibrio parahaemolyticus ATCC 17803, Escherichia coli O157:H7 ATCC 43894, and Pseudomonas aeruginosa KCTC 1637-at three different temperatures (4, 25, and 36 degrees C) with or without organic load (2% yeast extract). Results revealed a more than 4-log10 (CFU/ml) reduction (1.0 x 10(4) CFU/ml reduction) against all pathogens reacted at 37 degrees C for 3 h in the absence of organic material. GC-100X solution diluted with an equal volume of distilled or standard hard water (300 ppm CaCO3) showed effective bactericidal activity, particularly against gram-negative bacteria. Washing efficacy of GC-100X solution was compared against E. coli O157:H7 on cherry tomato surfaces with those of a commercially used detergent and chlorine water (100 ppm). Viable cell counts of E. coli O157:H7 that had penetrated to the cores of tomatoes after sanitizing treatment revealed that GC-100X stock and its 5% diluted solutions had similar washing effects to 100-ppm chlorine water and were more effective than the other kitchen detergent. These results indicate that GC-100X has good bactericidal and sanitizing activities and is useful as a new sanitizer for food safety and kitchen hygiene.     

Effectiveness of Individual or Combined Sanitizer Treatments for Inactivating Salmonella spp. on Smooth Surface, Stem Scar, and Wounds of Tomatoes

ABSTRACT: Unwaxed, green tomatoes ('Florida 47' cultivar) were contaminated with Salmonella and then treated with aqueous solutions of sodium hypochlorite (HOCl; 200 ppm), acidified sodium chlorite (ASC; 1200 ppm), peroxyacetic acid (PAA; 87 ppm), or chlorine dioxide gas (ClO₂; total 100 mg). Additionally, a combined treatment of immersion in HOCl, followed by immersion in ASC and then exposure to ClO₂ gas was investigated. Tomatoes were spot inoculated with a 5-strain Salmonella cocktail on smooth surfaces, stem scar tissue, or puncture wounds. A 3 replicate set of each of the sample groups was stored at 20 °C and 95% relative humidity (RH) and retested after 5 d. Greater than 4.0-log unit reductions of Salmonella spp. inoculated on the smooth surface of the tomatoes were seen for all aqueous sanitizer treatments, with Salmonella populations below the detection limit after 5 d of storage. All aqueous treatment groups showed > 1.0-log unit reductions in Salmonella at the stem scar and >2.0-log unit reduction at puncture wounds. The ClO₂ gas treatment reduced Salmonella to undetectable levels at the stem scar, but had no apparent effect on populations inoculated in puncture wounds. The combined treatment resulted in a 3.0-log unit reduction of inoculated Salmonella at puncture wounds. In all cases except for treatment with chlorine, surviving Salmonella populations did not increase after the 5 d of storage. Results of this study suggest the combined treatment was most effective for minimizing the risk of Salmonella contaminated on tomatoes.     

Reduction of Salmonella enterica contamination on grape tomatoes by washing with thyme oil, thymol, and carvacrol as compared with chlorine treatment

In recent years, multistate outbreaks of Salmonella enterica serovars were traced to tomatoes and resulted in serious economic loss for the tomato industry and decreased consumer confidence in the safety of tomato produce. Purified compounds derived from essential oils such as thymol and carvacrol had wide inhibitory effects against foodborne pathogens including Salmonella. The objective of this study was to determine the antimicrobial activities of thymol, carvacrol, and thyme oil against Salmonella on grape tomatoes. Surface-inoculated grape tomatoes were washed with 4% ethanol, 200 ppm of chlorine, or one of six washing solutions (thymol [0.2 and 0.4 mg/ml], thyme oil [1 and 2 mg/ml], and carvacrol [0.2 and 0.4 mg/ml]) for 5 or 10 min. There was no significant difference in the reduction of S. enterica serovars when different washing times were used (P > 0.05). Thymol (especially at the concentration of 0.4 mg/ml) was the most effective (P < 0.05) among the three natural antimicrobial agents, which achieved >4.1-log reductions of S. enterica serovars Typhimurium, Kentucky, Senftenberg, and Enteritidis on grape tomatoes after a 5-min washing and >4.3-log reductions after a 10-min washing. A >4.6-log reduction in the S. enterica populations in comparison to control was observed with the use of thymol solutions. The uses of these antimicrobial agents achieved significant log reductions of Salmonella on inoculated grape tomatoes and decreased dramatically the risk of potential transmission of pathogens from tomatoes to washing solutions. None of these antimicrobial agents decreased the total phenolic and ascorbic acid content, nor did any of them change the color and pH values or affect the taste, aroma, or visual quality of grape tomatoes. Therefore, 0.4 mg/ml thymol has great potential to be an alternative to chlorine-based washing solution for fresh produce.     

Efficiency of sodium hypochlorite, fumaric acid, and mild heat in killing native microflora and Escherichia coli O157:H7, Salmonella typhimurium DT104, and Staphylococcus aureus attached to fresh-cut lettuce

The effect of the disinfectant sodium hypochlorite (NaClO), with or without mild heat (50 degrees C) and fumaric acid, on native bacteria and the foodborne pathogens Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella Typhimurium DT104 attached to iceberg lettuce leaves was examined. The retail lettuce examined consistently harbored 6 to 7 log CFU/g of native bacteria throughout the study period. Inner leaves supported 1 to 2 log CFU/g fewer bacteria than outer leaves. About 70% of the native bacterial flora was removed by washing five times with 0.85% NaCl. S. aureus, E. coli, and Salmonella allowed to attach to lettuce leaves for 5 min were more easily removed by washing than when allowed to attach for 1 h or 2 days, with more S. aureus being removed than E. coli or Salmonella Typhimurium. An increase of time for attachment of pathogens from 5 min to 2 days leads to decreased efficiency of the washing and sanitizing treatment. Treatment with fumaric acid (50 mM for 10 min at room temperature) was the most effective, although it caused browning of the lettuce, with up to a 2-log reduction observed. The combination of 200 ppm of sodium hypochlorite and mild heat treatment at 50 degrees C for 1 min reduced the pathogen populations by 94 to 98% (1.2- to 1.7-log reduction) without increasing browning.     

Efficacy of neutral electrolyzed water (NEW) for reducing microbial contamination on minimally-processed vegetables

Consumption of minimally-processed, or fresh-cut, fruit and vegetables has rapidly increased in recent years, but there have also been several reported outbreaks associated with the consumption of these products. Sodium hypochlorite is currently the most widespread disinfectant used by fresh-cut industries. Neutral electrolyzed water (NEW) is a novel disinfection system that could represent an alternative to sodium hypochlorite. The aim of the study was to determine whether NEW could replace sodium hypochlorite in the fresh-cut produce industry. The effects of NEW, applied in different concentrations, at different treatment temperatures and for different times, in the reduction of the foodborne pathogens Salmonella, Listeria monocytogenes and Escherichia coli O157:H7 and against the spoilage bacterium Erwinia carotovora were tested in lettuce. Lettuce was artificially inoculated by dipping it in a suspension of the studied pathogens at 10(8), 10(7) or 10(5) cfu ml(-1), depending on the assay. The NEW treatment was always compared with washing with deionized water and with a standard hypochlorite treatment. The effect of inoculum size was also studied. Finally, the effect of NEW on the indigenous microbiota of different packaged fresh-cut products was also determined. The bactericidal activity of diluted NEW (containing approximately 50 ppm of free chlorine, pH 8.60) against E. coli O157:H7, Salmonella, L. innocua and E. carotovora on lettuce was similar to that of chlorinated water (120 ppm of free chlorine) with reductions of 1-2 log units. There were generally no significant differences when treating lettuce with NEW for 1 and 3 min. Neither inoculation dose (10(7) or 10(5) cfu ml(-1)) influenced the bacterial reduction achieved. Treating fresh-cut lettuce, carrot, endive, corn salad and 'Four seasons' salad with NEW 1:5 (containing about 50 ppm of free chlorine) was equally effective as applying chlorinated water at 120 ppm. Microbial reduction depended on the vegetable tested: NEW and sodium hypochlorite treatments were more effective on carrot and endive than on iceberg lettuce, 'Four seasons' salad and corn salad. The reductions of indigenous microbiota were smaller than those obtained with the artificially inoculated bacteria tested (0.5-1.2 log reduction). NEW seems to be a promising disinfection method as it would allow to reduce the amount of free chlorine used for the disinfection of fresh-cut produce by the food industry, as the same microbial reduction as sodium hypochlorite is obtained. This would constitute a safer, 'in situ', and easier to handle way of ensuring food safety.     

Evaluation of gaseous chlorine dioxide as a sanitizer for killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, and yeasts and molds on fresh and fresh-cut produce

Gaseous chlorine dioxide (ClO2) was evaluated for effectiveness in killing Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on fresh-cut lettuce, cabbage, and carrot and Salmonella, yeasts, and molds on apples, peaches. tomatoes, and onions. Inoculum (100 microl, ca. 6.8 log CFU) containing five serotypes of Salmonella enterica, five strains of E. coli O157:H7, or five strains of L. monocytogenes was deposited on the skin and cut surfaces of fresh-cut vegetables, dried for 30 min at 22 degrees C, held for 20 h at 4 degrees C, and then incubated for 30 min at 22 degrees C before treatment. The skin surfaces of apples, peaches, tomatoes, and onions were inoculated with 100 microl of a cell suspension (ca. 8.0 log CFU) containing five serotypes of Salmonella, and inoculated produce was allowed to dry for 20 to 22 h at 22 degrees C before treatment. Treatment with ClO2 at 4.1 mg/liter significantly (alpha = 0.05) reduced the population of foodborne pathogens on all produce. Reductions resulting from this treatment were 3.13 to 4.42 log CFU/g for fresh-cut cabbage, 5.15 to 5.88 log CFU/g for fresh-cut carrots, 1.53 to 1.58 log CFU/g for fresh-cut lettuce, 4.21 log CFU per apple, 4.33 log CFU per tomato, 1.94 log CFU per onion, and 3.23 log CFU per peach. The highest reductions in yeast and mold populations resulting from the same treatment were 1.68 log CFU per apple and 2.65 log CFU per peach. Populations of yeasts and molds on tomatoes and onions were not significantly reduced by treatment with 4.1 mg/liter ClO2. Substantial reductions in populations of pathogens on apples, tomatoes, and onions but not peaches or fresh-cut cabbage, carrot, and lettuce were achieved by treatment with gaseous ClO2 without markedly adverse effects on sensory qualities.     

Inactivation of Escherichia coli O157:H7, Salmonella enterica serotype enteritidis,and Listeria monocytogenes on lettuce by hydrogen peroxide and lactic acid and by hydrogen peroxide with mild heat

Iceberg lettuce is a major component in vegetable salad and has been associated with many outbreaks of foodborne illnesses. In this study, several combinations of lactic acid and hydrogen peroxide were tested to obtain effective antibacterial activity without adverse effects on sensory characteristics. A five-strain mixture of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis, and Listeria monocytogenes was inoculated separately onto fresh-cut lettuce leaves, which were later treated with 1.5% lactic acid plus 1.5% hydrogen peroxide (H2O2) at 40 degrees C for 15 min, 1.5% lactic acid plus 2% H2O2 at 22 degrees C for 5 min, and 2% H2O2 at 50 degrees C for 60 or 90 s. Control lettuce leaves were treated with deionized water under the same conditions. A 4-log reduction was obtained for lettuce treated with the combinations of lactic acid and H2O2 for E. coli O157:H7 and Salmonella Enteritidis, and a 3-log reduction was obtained for L. monocytogenes. However, the sensory characteristics of lettuce were compromised by these treatments. The treatment of lettuce leaves with 2% H2O2 at 50 degrees C was effective not only in reducing pathogenic bacteria but also in maintaining good sensory quality for up to 15 days. A < or = 4-log reduction of E. coli O157:H7 and Salmonella Enteritidis was achieved with the 2% H2O2 treatment, whereas a 3-log reduction of L. monocytogenes was obtained. There was no significant difference (P > 0.05) between pathogen population reductions obtained with 2% H2O2 with 60- and 90-s exposure times. Hydrogen peroxide residue was undetectable (the minimum level of sensitivity was 2 ppm) on lettuce surfaces after the treated lettuce was rinsed with cold water and centrifuged with a salad spinner. Hence, the treatment of lettuce with 2% H2O2 at 50 degrees C for 60 s is effective in initially reducing substantial populations of foodborne pathogens and maintaining high product quality.     

Calcinated calcium killing of Escherichia coli O157:H7, Salmonella,and Listeria monocytogenes on the surface of tomatoes

This study was conducted to evaluate the efficacy of calcinated calcium, 200 ppm chlorine water (1% active chlorine), and sterile distilled water in killing Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on the surfaces of spot-inoculated tomatoes. Inoculated tomatoes were sprayed with calcinated calcium, chlorinated water, or sterile distilled water (control) and hand rubbed for 30 s. Populations of E coli O157:H7, Salmonella, and L. monocytogenes in the rinse water and in the residual (0.1% peptone) wash solution were determined. Treatment with 200 ppm chlorine and calcinated calcium resulted in 3.40- and 7.85-log10 reductions of E. coli O157:H7, respectively, and 2.07- and 7.36-log10 reductions of Salmonella, respectively. Treatment with 200 ppm chlorine and calcinated calcium reduced L monocytogenes numbers by 2.27 and 7.59 log10 CFU per tomato, respectively. The findings of this study suggest that calcinated calcium could be useful in controlling pathogenic microorganisms in fresh produce.     

Inactivation of Escherichia coli O157:H7, Salmonella typhimurium DT104, and Listeria monocytogenes on inoculated alfalfa seeds with a fatty acid-based sanitizer

Alfalfa seeds were inoculated with a three-strain cocktail of Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium DT104, or Listeria monocytogenes by immersion to contain approximately 6 to 8 log CFU/g and then treated with a fatty acid-based sanitizer containing 250 ppm of peroxyacid, 1,000 ppm of caprylic and capric acids (Emery 658), 1,000 ppm of lactic acid, and 500 ppm of glycerol monolaurate at a reference concentration of 1X. Inoculated seeds were immersed at sanitizer concentrations of 5X, 10X, and 15X for 1, 3, 5, and 10 min and then assessed for pathogen survivors by direct plating. The lowest concentration that decreased all three pathogens by >5 log was 15. After a 3-min exposure to the 15X concentration, populations of E. coli O157:H7, Salmonella Typhimurium DT104, and L. monocytogenes decreased by >5.45, >5.62, and >6.92 log, respectively, with no sublethal injury and no significant loss in seed germination rate or final sprout yield. The components of this 15x concentration (treatment A) were assessed independently and in various combinations to optimize antimicrobial activity. With inoculated seeds, treatment C (15,000 ppm of Emery 658, 15,000 ppm of lactic acid, and 7,500 ppm of glycerol monolaurate) decreased Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes by 6.23 and 5.57 log, 4.77 and 6.29 log, and 3.86 and 4.21 log after 3 and 5 min of exposure, respectively. Treatment D (15,000 ppm of Emery 658 and 15,000 ppm of lactic acid) reduced Salmonella Typhimurium by >6.90 log regardless of exposure time and E. coli )157:H7 and L. monocytogenes by 4.60 and >5.18 log and 3.55 and 3.14 log after 3 and 5 min, respectively. No significant differences (P > 0.05) were found between treatments A, C, and D. Overall, treatment D, which contained Emery 658 and lactic acid as active ingredients, reduced E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes populations by 3.55 to >6.90 log and may provide a viable alternative to the recommended 20,000 ppm of chlorine for sanitizing alfalfa seeds.     

Effect of a simple chlorine dioxide method for controlling five foodborne pathogens, yeasts and molds on blueberries

The effect of aqueous chlorine dioxide (ClO(2)) on controlling foodborne pathogens, yeasts, and molds on blueberries was studied. Five pathogens were spot-inoculated on the skin of blueberries. A sachet containing necessary chemicals for generation of ClO(2) was used to provide 320 ppm of ClO(2) in 7.5 l of water. The efficacy of different concentrations (1, 3, 5, 10, and 15 ppm) of ClO(2) and various contact times (10s; 1, 5, 10, 20, 30 min; and 1h and 2h) were studied. ClO(2) was most effective in reducing Listeria monocytogenes (4.88 log cfu/g) as compared to the other pathogens. Pseudomonas aeruginosa was reduced by 2.16 log cfu/g after 5 min when treated with 15 ppm of ClO(2). Relatively short treatment time was more effective in reducing Salmonella Typhimurium than longer treatment time for most concentrations. The highest reduction (4.56 log cfu/g) of Staphylococcus aureus was achieved with 15 ppm of ClO(2) for 30 min. When treated for 2h with 5 ppm of ClO(2), Yersinia enterocolitica was reduced by 3.49 log cfu/g. Fifteen ppm of ClO(2) reduced natural yeasts and molds by 2.82 log cfu/g after 1h. Concentrations of ClO(2) decreased over time. When exposed to blueberries, ClO(2) concentrations were further reduced, showing significant degradation.     

Internalization of bacterial pathogens in tomatoes and their control by selected chemicals

The effect of different washing or sanitizing agents was compared for preventing or reducing surface and internal contamination of tomatoes by Salmonella Typhimurium and Escherichia coli O157:H7. The tomatoes were inoculated by dipping them in a bacterial suspension containing approximately 6.0 log CFU/ml of each pathogen and then rinsing them with tap water, hypochlorite solution (250 mg/liter), or lactic acid solution (2%, wt/vol). All treatments were applied by dipping or spraying, and solutions were applied at 5, 25, 35, and 55 degrees C. With the exception of the lactic acid dip at 5 degrees C, all treatments reduced both pathogens on the surfaces of the tomatoes by at least 2.9 cycles. No significantly different results were obtained (P > 0.05) with the dipping and spraying techniques. For internalized pathogens, the mean counts for tomatoes treated with water alone or with chlorine ranged from 0.8 to 2.1 log CFU/g. In contrast, after lactic acid spray treatment, all core samples of tomatoes tested negative for Salmonella Typhimurium and, except for one sample with a low but detectable count, all samples tested negative for E. coli O157:H7 with a plate count method. When the absence of pathogens was verified by an enrichment method, Salmonella was not recovered from any samples, whereas two of four samples tested positive for E. coli O157:H7 even though the counts were negative. Few cells of internalized pathogens were able to survive in the center of the tomato during storage at room temperature (25 to 28 degrees C). The average superficial pH of tomatoes treated with tap water, chlorine, or lactic acid was 4.9 to 5.2, 4.1 to 4.3, and 2.5, respectively (P < 0.05), whereas no differences were observed in the internal pH (3.6 to 3.7) of the tomatoes treated with different sanitizers. The general practice in the tomato industry is to wash the tomatoes in chlorinated water. However, chlorine is rapidly degraded by organic matter usually present in produce. Therefore, lactic acid sprays may be a more effective alternative for decontaminating tomato surfaces. The use of warm (55 degrees C) sprays could reduce pathogen internalization during washing.     

Combined effect of aqueous chlorine dioxide and modified atmosphere packaging on inhibiting Salmonella Typhimurium and Listeria monocytogenes in mungbean sprouts

This study was conducted to investigate the effect of chlorine dioxide (ClO2) combined with modified atmosphere packaging (MAP) on inhibiting total mesophilic microorganisms, Salmonella Typhimurium, and Listeria monocytogenes in mungbean sprouts during refrigerated storage. Mungbean sprouts were packaged using 4 different methods (air, vacuum, CO2 gas, and N2 gas) following treatment with water or 100 ppm ClO2 for 5 min and stored at 5 +/- 2 degrees C. The population of total mesophilic microorganisms in mungbean sprouts was about 8.4-log(10) CFU/g and this level was not significantly reduced by treatment with water or ClO2 (P > 0.05). However, when samples were packaged under vacuum, N2 gas, or CO2 gas following treatment with ClO2, the populations of total mesophilic microorganisms were significantly reduced during storage (P < 0.05). Levels of S. Typhimurium and L. monocytogenes in mungbean sprouts following inoculation were 4.6- and 5.6-log(10) CFU/g and treatment with water followed by different packaging conditions (air, vacuum, N2 gas, and CO2 gas) had no significant effect on population reduction (P > 0.05). However, treatment with ClO2 significantly reduced populations of S. Typhimurium and L. monocytogenes by 3.0- and 1.5-log CFU/g, respectively (P < 0.05), and these reduced cell levels were maintained or decreased in samples packaged under vacuum or in N2 or CO2 gas during storage. These results suggest that the combination of ClO2 treatment and MAP such as CO2 gas packaging may be useful for inhibiting microbial contamination and maintaining quality in mungbean sprouts during storage.     

High pressures in combination with antimicrobials to reduce Escherichia coli O157:H7 and Salmonella Agona in apple juice and orange juice

The effect of high pressure processing in conjunction with the chemical antimicrobials, dimethyl dicarbonate (DMDC), hydrogen peroxide, cinnamic acid, potassium sorbate, and sodium benzoate (NaB) on E. coli O157:H7 strain E009 and Salmonella enterica serovar Agona was investigated in apple juice and orange juice, respectively. Juices were inoculated with approximately 10(6) CFU/ml and subjected to pressures of 550 MPa (E. coli O157:H7 samples) and 400 MPa (Salmonella Agona samples) for 2 min at 6 degrees C (initial temperature). Populations of each pathogen were determined before pressurization, immediately after pressurization, and after samples had been held after treatment for 24 h at 4 degrees C. The most effective treatment for E. coli O157:H7, as determined by plating immediately after pressurization, was 125 ppm of DMDC, which caused a >4.98-log reduction. Other treatments that were significantly different from the sample with no added antimicrobial were 62.5 ppm of DMDC, 300 ppm of hydrogen peroxide, and 500 ppm of NaB, which produced 4.97-, 5.79-, and 3.91-log total reductions, respectively. After 24 h at 4 degrees C, E. coli O157:H7 was undetectable in all treatment groups (and controls). In samples inoculated with Salmonella, the most effective treatment was 62.5 ppm of DMDC, which produced a 5.96-log decrease immediately after pressure treatment. The results for 1,000 ppm of NaB, which produced a 3.26-log decrease, also were significantly different from those for the sample containing no antimicrobials. After 24 h at 4 degrees C, all samples with added antimicrobials had near or more than a 5-log total reduction of Salmonella Agona.     

High-pressure resistance variation of Escherichia coli O157:H7 strains and Salmonella serovars in tryptic soy broth, distilled water, and fruit juice

The effect of high pressure on the log reduction of six strains of Escherichia coli O157:H7 and five serovars of Salmonella enterica was investigated in tryptic soy broth, sterile distilled water, and commercially sterile orange juice (for Salmonella) and apple cider (for E. coli). Samples were subjected to high-pressure processing treatment at 300 and 550 MPa for 2 min at 6 degrees C. Samples were plated onto tryptic soy agar directly after pressurization and after being held for 24 h at 4 degrees C. At 300 MPa, little effect was seen on E. coli O157:H7 strains, while Salmonella serovars varied in resistance, showing reductions between 0.26 and 3.95 log CFU/ml. At 550 MPa, E. coli O157:H7 strains exhibited a range of reductions (0.28 to 4.39 log CFU/ml), while most Salmonella populations decreased beyond the detection limit (> 5-log CFU/ml reduction). The most resistant strains tested were E. coli E009 and Salmonella Agona. Generally, bacterial populations in fruit juices showed larger decreases than did populations in tryptic soy broth and distilled water. E. coli O157:H7 cultures held for 24 h at 4 degrees C after treatment at 550 MPa showed a significant log decrease as compared with cultures directly after treatment (P < or = 0.05), while Salmonella serovars did not show this significant decrease (P > 0.05). All Salmonella serovars tested in orange juice treated at 550 MPa for 2 min at 6 degrees C and held for 24 h showed a > 5-log decrease, while E. coli O157:H7 strains require a higher pressure, higher temperature, longer pressurization, or a chemical additive to achieve a 5-log decrease.     

Efficacy of chlorine dioxide gas as a sanitizer of lettuce leaves

Aqueous solutions of sodium hypochlorite or hypochlorous acid are typically used to sanitize fresh fruits and vegetables. However, pathogenic organisms occasionally survive aqueous sanitization in sufficient numbers to cause disease outbreaks. Chlorine dioxide (ClO2) gas generated by a dry chemical sachet was tested against foodborne pathogens on lettuce leaves. Lettuce leaves were inoculated with cocktail of three strains each of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium and treated with CLO2 gas for 30 min, 1 h, and 3 h in a model gas cabinet at room temperature (22 +/- 2 degrees C). After treatment, surviving cells, including injured cells, were enumerated on appropriate selective agar or using the overlay agar method, respectively. Total ClO2, generated by the gas packs was 4.3, 6.7, and 8.7 mg after 30 min, 1 h, and 3 h of treatment, respectively. Inoculated lettuce leaves exposed to ClO2 gas for 30 min experienced a 3.4-log reduction in E. coli, a 4.3-log reduction in Salmonella Typhimurium, and a 5.0-log reduction in L. monocytogenes when compared with the control. After 1 h. the three pathogens were reduced in number of CFU by 4.4. 5.3, and 5.2 log, respectively. After 3 h, the reductions were 6.9, 5.4, and 5.4 log, respectively. A similar pattern emerged when injured cells were enumerated. The ClO2, gas sachet was effective at killing pathogens on lettuce without deteriorating visual quality. Therefore, this product can be used during storage and transport of lettuce to improve its microbial safety.     

Response of Salmonella and Escherichia coli O157:H7 to UV energy

To determine the efficacy of a UV light treatment at 253.7 nm (UVC light) on microbial growth, plates containing tryptic soy agar plus 50 ppm of nalidixic acid (TSAN) were inoculated with known concentrations of five-strain cocktails of Salmonella and Escherichia coli O157:H7 and subjected to different UVC treatments. The concentration of the cocktail inoculum was determined with TSAN prior to inoculation. Serial dilutions were carried out, and inoculation levels of 10(0) to 10(8) CFU/ ml were tested for each pathogen. Multiple replications of doses of UV light ranging from 1.5 to 30 mW/cm2 were applied to different cocktail concentrations, and doses of > 8.4 mW/cm2 resulted in a 5-log reduction of Escherichia coli O157:H7, while a 5-log reduction of Salmonella was observed with doses of > 14.5 mW/cm2. Results for both organisms yielded sigmoidal inactivation curves. UVC light is effective in reducing microbial populations of pathogens on agar surfaces.     

Effects of cell surface charge and hydrophobicity on attachment of 16 Salmonella serovars to cantaloupe rind and decontamination with sanitizers

Adherence of bacteria to cantaloupe rind is favored by surface irregularities such as roughness, crevices, and pits, thus reducing the ability of washing or sanitizer treatments to remove or inactivate attached cells. In this study, we compared the surface charge and hydrophobicity of two cantaloupe-related outbreak strains of Salmonella Poona (RM2350 and G-91-1595) to those of 14 additional Salmonella strains using electrostatic and hydrophobic interaction chromatography. The relative abilities of the 16 strains to attach to cantaloupe surfaces and resist removal by washing with water, chlorine (200 ppm), or hydrogen peroxide (2.5%) for 5 min after a storage period of up to 7 days at 5 to 20 degrees C also were determined. Whole cantaloupes were inoculated with each pathogen at 8.36 log CFU/ml, dried for 1 h inside a biosafety cabinet, stored, and then subjected to the washing treatments. Only the positive surface charge of the two cantaloupe-related strains of Salmonella Poona was significantly higher (P < 0.05) than that of the other strains. Initial bacterial attachment to cantaloupe surfaces ranged from 3.68 to 4.56 log CFU/cm2 (highest values for Salmonella Michigan, Newport, Oranienburg, and Mbandaka). The average percentage of the total bacterial population strongly attached to the cantaloupe surface for the Salmonella serovars studied ranged from 0.893 to 0.946 at 5 degrees C and from 0.987 to 0.999 at 25 degrees C. Washing inoculated melons with water did not produce a significant reduction in the concentration of the pathogens (P > 0.05). Chlorine and hydrogen peroxide treatments caused an average 3-log reduction when applied 20 to 40 min postinoculation. However, sanitizer treatments applied 60 min or more postinoculation were less effective (approximately 2.5-log reduction). No significant differences were noted in sanitizer efficacy against the individual strains (P > 0.05). The two cantaloupe-related outbreak Salmonella Poona strains did not significantly differ from the other Salmonella strains tested in negative cell surface charge or hydrophobicity, were not more effective in attaching to whole melon surfaces, and were not more resistant to the various washing treatments when present on rinds.     

Effects of tomato variety, temperature differential, and post-stem removal time on internalization of Salmonella enterica serovar Thompson in tomatoes

Tomatoes have been implicated in salmonellosis outbreaks due to possible contamination through bacterial internalization during postharvest handling. This study was conducted to determine the effects of tomato variety, temperature differential between tomato pulp and bacterial suspension, and the time delay between stem removal and immersion in bacterial suspension on internalization of Salmonella enterica serovar Thompson in tomato fruit. Mature green tomatoes at 32.2°C were immersed in water containing approximately 10(6) CFU/ml S. enterica bacteria. Different tomato varieties (Mountain Spring, Applause, and BHN961), temperature differentials (-10, 0, and 10°F, or -5.6, 0, and 5.6°C, respectively), and post-stem removal times (0, 2, and 16 h) were evaluated for their effects on S. enterica internalization. The incidence and density of internalized cells were determined by culture enrichment and most-probable-number methods, respectively. Overall, variety and post-stem removal time by variety interaction significantly affected the incidence of S. enterica internalization (P < 0.0001), while temperature differential had no significant effect (P = 0.36). Mountain Spring tomatoes were less susceptible to S. enterica internalization than were Applause and BHN961. Increasing the time interval between stem removal and immersion greatly reduced pathogen internalization in BHN961 and Applause, while it had no effect in Mountain Spring tomatoes. The variety and interactions between varieties and post-stem removal times (P = 0.0363) and between temperature differentials and post-stem removal times (P = 0.0257) had significant effects on the populations of internalized S. enterica. Furthermore, all internalized S. enterica cells were found within the core tissue segments immediately underneath the stem scars.     

Determination of 5-log reduction times for food pathogens in acidified cucumbers during storage at 10 and 25 degrees C

Outbreaks of acid-resistant foodborne pathogens in acid foods with pH values below 4.0, including apple cider and orange juice, have raised concerns about the safety of acidified vegetable products. For acidified vegetable products with pH values between 3.3 and 4.6, previous research has demonstrated that thermal treatments are needed to achieve a 5-log reduction in the numbers of Escherichia coli O157:H7, Listeria monocytogenes, or Salmonella enterica. For some acidified vegetable products with a pH of 3.3 or below, heat processing can result in unacceptable product quality. The purpose of this study was to determine the holding times needed to achieve a 5-log reduction in E. coli O157:H7, L. monocytogenes, and S. enterica strains in acidified vegetable products with acetic acid as the primary acidulant, a pH of 3.3 or below, and a minimum equilibrated temperature of 10 degrees C. We found E. coli O157:H7 to be the most acid-resistant microorganism for the conditions tested, with a predicted time to achieve a 5-log reduction in cell numbers at 10 degrees C of 5.7 days, compared with 2.1 days (51 h) for Salmonella or 0.5 days (11.2 h) for Listeria. At 25 degrees C, the E. coli O157:H7 population achieved a 5-log reduction in 1.4 days (34.3 h).     

The Effectiveness of Sanitizer Treatments in Inactivation of Salmonella spp. from Bell Pepper, Cucumber, and Strawberry

ABSTRACT: Four different postharvest treatments for removal of Salmonella from bell pepper and cucumber were examined, including washes with chlorinated water (HOCl; 200 ppm), acidified sodium chlorite (ASC; 1200 ppm), and peroxyacetic acid (PAA; 75 ppm), and treatment with gaseous chlorine dioxide (ClO₂; total 100 mg). Only ClO₂ gas was evaluated for decontamination of strawberries. Each produce was inoculated with approximately 1.0 × 10⁷ colony-forming units (CFU) of a 5-serovar cocktail of Salmonella on artificially created wounds, smooth surfaces, and stem scar tissue. For tests involving smooth surface inoculation, ASC and PAA treatments decreased contamination to undetectable levels on bell pepper and cucumber, while the chlorine treatment of bell pepper reduced contamination by approximately 2-logs. For stem scar contamination on bell pepper, ASC and PAA treatments both showed >2-log unit reductions, and chlorine treatment showed a <1-log unit reduction. For puncture wounds on bell pepper, HOCl, ASC, and PAA treatments reduced bacterial levels approximately 2-, 3-, and 1-log units, respectively, indicating that HOCl and ASC were more effective than PAA. These aqueous treatments of cucumber with puncture wounds reduced bacterial levels approximately 1-, 2-, and 2-log units, respectively. ClO₂ treatment decreased counts to undetectable levels on all inoculation sites on cucumber and on strawberry smooth surfaces, but failed to completely eliminate Salmonella from bell pepper and from the stem scar and the puncture wounds of strawberry. ASC treatment of bell pepper and ClO₂ gas treatments of cucumber showed the best efficiency for inactivation of Salmonella. ClO₂ treatments effectively reduced Salmonella cells inoculated on the smooth surface and stem scar of strawberries compared with unsanitized control.