Efficacy of Neutral pH Electrolyzed Water in Reducing Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 on Fresh Produce Items using an Automated Washer at Simulated Food Service Conditions

The objective of this study was to determine the efficacy of neutral pH electrolyzed (NEO) water (155 mg/L free chlorine, pH 7.5) in reducing Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 on romaine lettuce, iceberg lettuce, and tomatoes washed in an automated produce washer for different times and washing speeds. Tomatoes and lettuce leaves were spot inoculated with 100 μL of a 5 strain cocktail mixture of either pathogen and washed with 10 or 8 L of NEO water, respectively. Washing lettuce for 30 min at 65 rpm led to the greatest reductions, with 4.2 and 5.9 log CFU/g reductions achieved for E. coli O157:H7 and S. Typhimurium respectively on romaine, whereas iceberg lettuce reductions were 3.2 and 4.6 log CFU/g for E. coli O157:H7 and S. Typhimurium respectively. Washing tomatoes for 10 min at 65 rpm achieved reductions greater than 8 and 6 log CFU/tomato on S. Typhimurium and E. coli O157:H7 respectively. All pathogens were completely inactivated in NEO water wash solutions. No detrimental effects on the visual quality of the produce studied were observed under all treatment conditions. Results show the adoption of this washing procedure in food service operations could be useful in ensuring produce safety.     

Switchgrass extractives to mitigate Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium contamination of romaine lettuce at pre- and postharvest

The antimicrobial potential of switchgrass extractives (SE) was evaluated on cut lettuce leaves and romaine lettuce in planta, using rifampicin-resistant Escherichia coli O157:H7 and Salmonella Typhimurium strain LT2 as model pathogens. Cut lettuce leaves were swabbed with E. coli O157:H7 or S. Typhimurium followed by surface treatment with 0.8% SE, 0.6% sodium hypochlorite, or water for 1 to 45 min. For in planta studies, SE was swabbed on demarcated leaf surfaces either prior to or after inoculation of greenhouse-grown lettuce with E. coli O157:H7 or S. Typhimurium; the leaf samples were collected after 0, 24, and 48 h of treatment. Bacteria from inoculated leaves were enumerated on tryptic soy agar plates (and also on MacConkey's and XLT4 agar plates), and the recovered counts were statistically analyzed. Cut lettuce leaves showed E. coli O157:H7 reduction between 3.25 and 6.17 log CFU/leaf, whereas S. Typhimurium reductions were between 2.94 log CFU/leaf and 5.47 log CFU/leaf depending on the SE treatment durations, from initial levels of ∼7 log CFU/leaf. SE treatment of lettuce in planta, before bacterial inoculation, reduced E. coli O157:H7 and S. Typhimurium populations by 1.88 and 2.49 log CFU after 24 h and 3 h, respectively. However, SE treatment after bacterial inoculation of lettuce plants decreased E. coli O157:H7 populations by 3.04 log CFU (after 0 h) with negligible reduction of S. Typhimurium populations. Our findings demonstrate the potential of SE as a plant-based method for decontaminating E. coli O157:H7 on lettuce during pre- and postharvest stages in hurdle approaches.     

Efficacy of slightly acidic electrolyzed water in killing or reducing Escherichia coli O157:H7 on iceberg lettuce and tomatoes under simulated food service operation conditions

Abstract: The objective of this study was to evaluate the efficacy of slightly acidic electrolyzed (SAEO) water in killing or removing Escherichia coli O157:H7 on iceberg lettuce and tomatoes by washing and chilling treatment simulating protocols used in food service kitchens. Whole lettuce leaves and tomatoes were spot‐inoculated with 100 μL of a mixture of 5 strains of E. coli O157:H7. Washing lettuce with SAEO water for 15 s reduced the pathogen by 1.4 to 1.6 log CFU/leaf, but the treatments did not completely inactivate the pathogen in the wash solution. Increasing the washing time to 30 s increased the reductions to 1.7 to 2.3 log CFU/leaf. Sequential washing in SAEO water for 15 s and then chilling in SAEO water for 15 min also increased the reductions to 2.0 to 2.4 log CFU/leaf, and no cell survived in chilling solution after treatment. Washing tomatoes with SAEO water for 8 s reduced E. coli O157:H7 by 5.4 to 6.3 log CFU/tomato. The reductions were increased to 6.6 to 7.6 log CFU/tomato by increasing the washing time to 15 s. Results suggested that application of SAEO water to wash and chill lettuce and tomatoes in food service kitchens could minimize cross‐contamination and reduce the risk of E. coli O157:H7 present on the produce. Practical Application: SAEO water is equally or slightly better than acidic electrolyzed (AEO) water for inactivation of bacteria on lettuce and tomato surfaces. In addition, SAEO water may have the advantages over AEO water on its stability, no chlorine smell, and low corrosiveness. Therefore, SAEO water may have potential for produce wash to enhance food safety.     

Efficacy of Chlorine Dioxide, Ozone, and Thyme Essential Oil or a Sequential Washing in Killing Escherichia coli O157:H7 on Lettuce and Baby Carrots

Chlorine dioxide (ClO₂), ozone, and thyme essential oil has been found to be effective in reducing pathogens, including Escherichia coli O157:H7, on selected produce. The efficacy of these sanitizers was evaluated, alone or through their sequential washing to achieve a 3 or more log reduction of mixed strains of E. coli O157:H7 on shredded lettuce and baby carrots. Samples sprinkle inoculated with mixed strains of E. coli O157:H7 were air-dried for 1 h at 22±2°C in a biosafety cabinet, stored at 4°C for 24 h, and then treated with different concentrations of disinfectants and exposure time. Sterile deionized water washing resulted in approximately 1log reduction ofE. coli O157:H7 after 10 min washing of lettuce and baby carrots. Gaseous treatments resulted in higher log reductions in comparison to aqueous washing. However, decolorization of lettuce leaves was observed during long exposure time. A logarithmic reduction of 1.48-1.97log₁₀ cfu/g was obtained using aqueous ClO₂ (10.0 mg/L for 10 min) ozonated water (9.7 mg/L for 10 min) or thyme oil suspension (1.0 mL/L for 5 min) on lettuce and baby carrots. Of the three sequential washing treatments used in this study, thyme oil followed by aqueous ClO₂/ozonated water, or ozonated water/aqueous ClO₂ were significantly (P<0.05) more effective in reducing E. coli O157:H7 (3.75 and 3.99log, and 3.83 and 4.34 log reduction) on lettuce and baby carrots, respectively. The results obtained from this study indicate that sequential washing treatments could achieve 3-4log reduction of E. coli O157:H7 on shredded lettuce and baby carrots.     

The Effect of pH on Inactivation of Pathogenic Bacteria on Fresh-cut Lettuce by Dipping Treatment with Electrolyzed Water

Fresh‐cut lettuce samples inoculated with S. Typhimurium, E. coli O157:H7 or L. monocytogenes were dipped into 300 ppm electrolyzed water (EW) at pH 4 to 9 and 30 °C for 5 min. The effects of treatment pH on bacterial reduction and visual quality of the lettuce were determined. The treatments at pH 4 and 8 resulted in the most effective inactivation of E. coli O157:H7, but the effect of pH was not significant (P > 0.05) for S. Typhimurium and L. monocytogenes. The treatment at pH 7 retained the best visual quality of lettuce, and achieved a reduction of approximately 2 log CFU/g for above 3 bacteria.     

Inactivation of Escherichia coli O157:H7, Salmonella typhimurium,and Listeria monocytogenes on Stored Iceberg Lettuce by Aqueous Chlorine Dioxide Treatment

ABSTRACT: Inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes in iceberg lettuce by aqueous chlorine dioxide (ClO₂) treatment was evaluated. Iceberg lettuce samples were inoculated with approximately 7 log CFU/g of E. coli O157:H7, S. typhimurium, and L. monocytogenes. Iceberg lettuce samples were then treated with 0, 5, 10, or 50 ppm ClO₂ solution and stored at 4 °C. Aqueous ClO₂ treatment significantly decreased the populations of pathogenic bacteria on shredded lettuce (P < 0.05). In particular, 50 ppm ClO₂ treatment reduced E. coli O157:H7, S. typhimurium, and L. monocytogenes by 1.44, 1.95, and 1.20 log CFU/g, respectively. The D₁₀‐values of E. coli O157:H7, S. typhimurium, and L. monocytogenes in shredded lettuce were 11, 26, and 42 ppm, respectively. The effect of aqueous ClO₂ treatment on the growth of pathogenic bacteria during storage was evaluated, and a decrease in the population size of these pathogenic bacteria was observed. Additionally, aqueous ClO₂ treatment did not affect the color of lettuce during storage. These results suggest that aqueous ClO₂ treatment can be used to improve the microbial safety of shredded lettuce during storage.     

Transfer of Escherichia coli O157:H7 to romaine lettuce due to contact water from melting ice

Ice can be used to chill romaine lettuce and maintain relative humidity during transportation. Escherichia coli O157:H7 may contaminate water used for ice. The objective of this study was to determine the potential for E. coli O157:H7 contamination of romaine lettuce from either ice contaminated with the pathogen or by transfer from lettuce surfaces via melting ice. In experiment 1, lettuce was spot inoculated with E. coli O157:H7 and chilled with ice prepared from uncontaminated tap water. In experiment 2, water inoculated with this pathogen was frozen and used to ice lettuce. Three heads of lettuce were stacked in each container and stored at 4 or 20 degrees C. After the ice melted, E. coli O157:H7 attachment to and recovery from the lettuce leaves were determined. For experiment 1, the population of E. coli O157:H7 attached to inoculated sites averaged 3.8 and 5.5 CFU/cm2 at 4 and 20 degrees C, respectively. Most of the uninoculated sites became contaminated with the pathogen due to ice melt. For experiment 2, 3.5 to 3.8 log CFU E. coli O157:H7 per cm2 was attached to the top leaf on the first head. After rinsing with chlorinated water (200 microg/ml), E. coli O157:H7 remained on the surface of the top head (1.8 to 2.0 log CFU/cm2). There was no difference in numbers of E. coli O157:H7 recovered from each sampling site at 4 and 20 degrees C. Results show that E. coli O157:H7 can be transferred onto other produce layers in shipping containers from melted ice made of contaminated water and from contaminated to uncontaminated leaf surfaces.     

Edible films containing carvacrol and cinnamaldehyde inactivate Escherichia coli O157:H7 on organic leafy greens in sealed plastic bags

The antimicrobial effects of apple-, carrot-, and hibiscus-based edible films containing carvacrol and cinnamaldehyde against Escherichia coli O157:H7 on organic leafy greens in sealed plastic bags were investigated. Fresh-cut Romaine and Iceberg lettuce, and mature and baby spinach leaves were inoculated with E. coli O157:H7 and placed into Ziploc® bags. Edible films were then added to the bags, which were stored at 4°C. The evaluation of samples taken at days 0, 3, and 7 showed that on all leafy greens, 3% carvacrol-containing films had the greatest effect against E. coli O157:H7, reducing the bacterial population by about 5 log CFU/g on day 0. All three types of 3% carvacrol-containing films reduced E. coli O157:H7 by about 5 log CFU/g at day 0. The 1.5% carvacrol-containing films reduced E. coli O157:H7 by 1–4 logs CFU/g at day 7. Films with 3% cinnamaldehyde showed reduction of 0.6–3 logs CFU/g on different leafy greens.     

Fate of Escherichia coli O157:H7 in field-inoculated lettuce

Impact of drip and overhead sprinkler irrigation on the persistence of attenuated Escherichia coli O157:H7 in the lettuce phyllosphere was investigated using a split-plot design in four field trials conducted in the Salinas Valley, California, between summer 2007 and fall 2009. Rifampicin-resistant attenuated E. coli O157:H7 ATCC 700728 (BLS1) was inoculated onto the soil beds after seeding with a backpack sprayer or onto 2- or 4-week-old lettuce plant foliage with a spray bottle at a level of 7 log CFU ml⁻¹. When E. coli O157:H7 was inoculated onto 2-week-old plants, the organism was recovered by enrichment in 1 of 120 or 0 of 240 plants at 21 or 28 days post-inoculation, respectively. For the four trials where inoculum was applied to 4-week-old plants, the population size of E. coli O157:H7 declined rapidly and by day 7, counts were near or below the limit of detection (10 cells per plant) for 82% or more of the samples. However, in 3 out 4 field trials E. coli O157:H7 was still detected in lettuce plants by enrichment 4-weeks post-inoculation. Neither drip nor overhead sprinkler irrigation consistently influenced the survival of E. coli O157:H7 on lettuce.     

Comparison of the Microbiological Quality of Environmentally Friendly and Conventionally Grown Vegetables Sold at Retail Markets in Korea

Fresh produce is usually eaten raw without cooking or heating, which may increase the probability of foodborne infection. The microbiological quality of 11 types of fresh, raw vegetables (romaine lettuce, sesame leaves, crown daisy, garlic chives, iceberg lettuce, cabbage, broccoli, leek, chili pepper, capsicum, and zucchini) purchased at retail markets in Iksan, Korea as affected by cultivation method (environmentally friendly vegetables [organic, pesticide‐free, and low‐pesticide vegetables] and conventionally grown vegetables) and harvest season was determined. Escherichia coli O157:H7 and Salmonella were not detected in all samples of vegetables tested. Aerobic mesophiles (>6 log cfu/g) were detected in environmentally friendly romaine lettuce and crown daisy and environmentally friendly and conventionally grown garlic chives, which also contained coliforms (>3 log cfu/g). Sesame leaf and crown daisy (regardless of cultivation method), as well as conventionally grown romaine lettuce and leek, contained >1 log cfu/g of E. coli. The overall microbiological quality of environmentally friendly and conventionally grown vegetables was not significantly different (P > 0.05). However, there were seasonal effects on populations of coliforms and generic E. coli on vegetables. The greatest numbers of microorganisms were isolated from environmentally friendly or conventionally grown vegetables purchased in winter. The vegetables, regardless of cultivation method or season, should be subjected to appropriate antimicrobial treatment to enhance their microbial safety.     

Effectiveness of Active Packaging on Control of Escherichia Coli O157:H7 and Total Aerobic Bacteria on Iceberg Lettuce

Contaminated leafy green vegetables have been linked to several outbreaks of human gastrointestinal infections. Antimicrobial interventions that are adoptable by the fresh produce industry for control of pathogen contamination are in great demand. This study was undertaken to evaluate the efficacy of sustained active packaging on control of Escherichia coli O157:H7 and total aerobic bacteria on lettuce. Commercial Iceberg lettuce was inoculated with a 3‐strain mixture of E. coli O157:H7 at 10² or 10⁴ CFU/g. The contaminated lettuce and un‐inoculated controls were placed respectively in 5 different active packaging structures. Traditional, nonactive packaging structure was included as controls. Packaged lettuce was stored at 4, 10, or 22 °C for 3 wk and sampled weekly for the population of E. coli O157:H7 and total aerobic bacteria. Results showed that packaging structures with ClO₂ generator, CO₂ generator, or one of the O₂ scavengers effectively controlled the growth of E. coli O157:H7 and total aerobic bacteria under all storage conditions. Packaging structure with the ClO₂ generator was most effective and no E. coli O157:H7 was detected in samples packaged in this structure except for those that were inoculated with 4 log CFU/g of E. coli O157:H7 and stored at 22 °C. Packaging structures with an oxygen scavenger and the allyl isothiocyanate generator were mostly ineffective in control of the growth of the bacteria on Iceberg lettuce. The research suggests that some of the packaging structures evaluated in the study can be used to control the presence of foodborne pathogens on leafy green vegetables.     

Efficacy of Ozone Against Escherichia coli O157:H7 on Apples

Apples were inoculated with Escherichia coli O157:H7 and treated with ozone. Sanitization treatments were more effective when ozone was bubbled during apple washing than by dipping apples in pre‐ozonated water. The corresponding decreases in counts of E. coli O157:H7 during 3‐min treatments were 3.7 and 2.6 log₁₀ CFU on apple surface, respectively, compared to < 1 log₁₀ CFU decrease in the stem‐calyx region in both delivery methods. Optimum conditions for decontamination of whole apples with ozone included a pretreatment with a wetting agent, followed by bubbling ozone for 3 min in the wash water, which decreased the count of E. coli O157:H7 by 3.3 log₁₀CFU/g.     

Survival of Escherichia coli O157:H7 in channel catfish pond and holding tank water

Survival of Escherichia coli O157:H7 in channel catfish (Ictalurus punctatus), pond and holding tank water was investigated. Water from three channel catfish ponds was inoculated with ampicillin/nalidixic acid-resistant E. coli O157:H7 transformed with a plasmid encoding for green fluorescent protein at 10⁵, 10⁶, and 10⁷ CFU/ml. Samples were taken from surface, internal organs, and skin scrape of fish and pond water for E. coli O157:H7 enumeration on brain heart infusion (BHI) agar containing ampicillin and nalidixic acid. To determine the survival of E. coli O157:H7 in catfish holding tank water from two farmers markets, the water was inoculated with 10⁷E. coli O157:H7 CFU/ml. E. coli O157:H7 were detected by direct plating for 33 and 69 d in pond and holding tank water, respectively. A rapid decrease of the pathogen was observed in the first 2 weeks to reach 2 log CFU/ml. When E. coli O157:H7 was not recovered by direct plating, the pathogen was isolated by enrichment in TSB for approximately another 30 d from pond and holding tank water. The populations of E. coli O157:H7 found in the internal organs and skin scrape were 5.5 log and 2.5 log CFU/ml, respectively. E. coli O157:H7 from internal organs and water were recovered for at least 12 d. Results suggest that E. coli O157:H7 can survive in channel catfish pond and holding tank water and channel catfish may become a potential carrier of the pathogen.     

Decontamination of irrigation water using a combined sand filtration and UV-C light treatment

Surface irrigation water is a food safety risk due to susceptibility to contamination and varying turbidity that affects chlorination efficacy. Effectiveness of a combined high-throughput, commercially available sand filter and UV-C system to inactivate Escherichia coli (K12, and attenuated O157:H7) in simulated irrigation water of varying turbidity and actual irrigation water was investigated. Sand filtration reduced the water turbidity from ~600 nephelometric turbidity units (NTU) to ~200 NTU but did not effectively filter out E. coli (K12) [<1 log colony forming units (CFU)/ml]. UV-C-induced inactivation of E. coli (attenuated O157:H7) was modeled for various levels of turbidity (0–500 NTU), and the relationship between the D value for inactivation and turbidity was found to be linear (R² > .97). Predicted UV treatment time to achieve 5 log reduction in water with turbidities of 100 and 300 NTU was 110 and 370 s, respectively. Validation performed with irrigation water (turbidity of 330 ± 82 NTU) showed that sand filtration reduced turbidity to 180 ± 57 NTU. Based on this, the duration of UV treatment required for 5 log reduction of E. coli O157:H7 was calculated (average of 237 s). Observed levels of inactivation, 4.7 ± 0.31 log CFU/ml, were not significantly different (p > .05) from the predicted level, 5 log CFU/ml, indicating a good model fit.     

Survival and growth of foodborne pathogens in minimally processed vegetables at 4 and 15 °C

Abstract: We conducted this study to investigate the survival and growth of pathogens on fresh vegetables stored at 4 and 15 °C. Vegetables (romaine lettuce, iceberg lettuce, perilla leaves, and sprouts) were inoculated with 4 pathogens (Salmonella enterica serovar Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Escherichia coli O157:H7) and stored at 2 different temperatures for different periods of time (3, 6, 9, 12, and 15 d at 4 °C and 1, 2, 3, 5, and 7 d at 15 °C). Populations of the 4 pathogens tended to increase on all vegetables stored at 15 °C for 7 d. Populations of E. coli O157:H7 and S. Typhimurium increased significantly, by approximately 2 log₁₀CFU/g, on loose and head lettuce stored at 15 °C for 1 d. No significant differences were observed in the growth of different pathogens on vegetables stored at 4 °C for 15 d. E. coli O157:H7 did not survive on sprouts stored at 15 or 4 °C. The survival and growth of food pathogens on fresh vegetables were very different depending on the pathogen type and storage temperature. Practical Application: Survivals and growth of pathogens on various vegetables at 4 and 15 °C were observed in this study. Survivals and growth of pathogens on vegetables were different depending on the pathogen type and storage temperature. Therefore, vegetables should be stored under refrigerated conditions (below 4 °C) prior to consumption. This recommendation may vary depending on the type of vegetable.     

Effects of X-ray radiation on Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri inoculated on shredded iceberg lettuce

The main goal of this investigation was to study the efficacy of X-ray doses (0.1, 0.2, 0.3, 0.5, 0.75, 1.0, 1.5 and 2.0 kGy) on inoculated Escherichia coli O157: H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on shredded iceberg lettuce. The second goal was to study the effect of X-ray on the inherent microflora counts and visual color of shredded iceberg lettuce during storage at 4 °C for 30 days. Treatment with 1.0 kGy X-ray significantly reduced the population of E. coli O157: H7, L. monocytogenes, Salmonella enterica and S. flexneri on shredded iceberg lettuce by 4.4, 4.1, 4.8 and 4.4-log CFU 5 cm⁻², respectively. Furthermore, more than a 5 log CFU reduction of E. coli O157: H7, L. monocytogenes, S. enterica and S. flexneri was achieved with 2.0 kGy X-ray. Treatment with X-ray reduced the initial microflora on iceberg lettuce and kept them significantly (p < 0.05) lower than the control during storage at 4 °C and 90% RH for 30 days. Treatment with X-ray did not significantly (p > 0.05) change the green color of iceberg lettuce leaves. Treatment with X-ray significantly reduced selected pathogens and inherent microorganisms on shredded iceberg lettuce leaves, which could be a good alternative to other technologies for produce (lettuce) industry.     

Role of O-antigen on the Escherichia coli O157:H7 cells hydrophobicity, charge and ability to attach to lettuce

Environmental factors encountered during growing and harvesting may contribute to Escherichia coli O157:H7 contamination of lettuce. Limited nutrients and extended exposure to water may cause E. coli O157:H7 to shed its O antigen. Absence of the O157-polysaccharide antigen could affect the cell's physicochemical properties (hydrophobicity and cell charge) and ultimately influence its attachment to surfaces. The objectives of this study were to evaluate the effect of the E. coli O157:H7 O-antigen on the cell's overall hydrophobicity, charge and ability to attach to cut edge and whole leaf iceberg lettuce surfaces. Three strains of E. coli O157:H7 (86-24 wild type; F-12, mutant lacking the O-antigen and pRFBE, plasmid for O157 gene reintroduced) were examined for their hydrophobicity, overall charge and ability to attach to lettuce. Overall, E. coli O157:H7 attached at higher levels to cut surfaces over whole leaf surfaces (P = 0.008) for all strains and treatments. Additionally, the strain lacking the O-antigen (F12) — attached significantly less to lettuce (P = 0.015) than the strains expressing the antigen (WT and pRFBE). Cells lacking the O antigen (strain F-12) were also significantly more hydrophobic than strains 86-24 or pRFBE (P ≤ 0.05). Surface charge differed among the strains tested (P ≤ 0.05); however, it did not appear to influence bacterial attachment to lettuce surfaces. The charge was not fully restored in the pRFBE strain (expression of O-antigen reintroduced), therefore, no conclusions can be made pertaining to the effect of charge on attachment in this study. Results indicate that E. coli O157:H7 cells which lack the O-antigen have greater hydrophobicity and attach at lower concentrations than cells expressing the O-antigen, to iceberg lettuce surfaces.     

Reduction of Escherichia coli O157:H7 in Biofilms Using Bacteriophage BPECO 19

Biofilm formation is a growing concern in the food industry. Escherichia coli O157:H7 is one of the most important foodborne pathogens that can persists in food and food‐related environments and subsequently produce biofilms. The efficacy of bacteriophage BPECO 19 was evaluated against three E. coli O157:H7 strains in biofilms. Biofilms of the three E. coli O157:H7 strains were grown on abiotic (stainless steel, rubber, and minimum biofilm eradication concentration [MBEC^TM] device) and biotic (lettuce) surfaces at different temperatures. The effectiveness of bacteriophage BPECO 19 in reducing preformed biofilms on these surfaces was further evaluated by treating the surfaces with a phage suspension (10⁸ PFU/mL) for 2 h. The results indicated that the phage treatment significantly reduced (P  < 0.05) the number of adhered cells in all the surfaces. Following phage treatment, the viability of adhered cells was reduced by ≥3 log CFU/cm², 2.4 log CFU/cm², and 3.1 log CFU/peg in biofilms grown on stainless steel, rubber, and the MBEC^TM device, respectively. Likewise, the phage treatment reduced cell viability by ≥2 log CFU/cm² in biofilms grown on lettuce. Overall, these results suggested that bacteriophages such as BPECO 19 could be effective in reducing the viability of biofilm‐adhered cells.     

Detection of 5 CFU/g of Escherichia coli O157:H7 on lettuce using activated charcoal and real-time PCR without enrichment

A sample treatment method which separates Escherichia coli O157:H7 from lettuce and removes PCR inhibitors allowing 5 CFU/g of target cells to be detected using real-time PCR is described. Lettuce leaves inoculated with E. coli O157:H7 were rinsed with 0.025% sodium dodecyl sulfate (SDS). In this study, there were two major factors that strongly affected the recovery of E. coli O157:H7 during sample preparation, the amount of bentonite coated activated charcoal used to remove PCR inhibitors and the agitated contact time of the samples with the coated charcoal. When 3.0 g of activated carbon coated with bentonite were mixed with target cell suspensions (30 ml) derived from 50 g of lettuce, a high recovery of E. coli O157:H7 (93%) was obtained. Sample agitation with bentonite coated activated charcoal for 15 min resulted in 95% recovery of E. coli O157:H7. When a commercial DNA purification resin was used for detection of E. coli O157:H7 without the use of the bentonite treated charcoal, the real-time PCR (Rti-PCR) failed to detect 1 × 10² CFU/g. In contrast, with the use of use of bentonite coated activated charcoal and a commercial DNA purifying resin together, Rti-PCR was able to detect 5 CFU of E. coli O157:H7/g of lettuce which was equivalent to 2.8 CFU/Rti-PCR. Such a successful detection level was the result of the bentonite coated activated charcoal’s ability to absorb the PCR inhibitors released from seeded lettuce during detachment. A standard curve was generated by plotting the Ct values against the log of CFU of target bacterial cells. A linear range of DNA amplification was exhibited from 5.0 × 10⁰ to 1.0 × 10⁴ CFU/g by using Rti-PCR.     

Efficacy of adding detergents to sanitizer solutions for inactivation of Escherichia coli O157:H7 on Romaine lettuce

Numerous Escherichia coli O157:H7 outbreaks have been linked to consumption of fresh lettuce. The development of effective and easily implemented wash treatment could reduce such incidents. The purpose of this study was to evaluate the addition of food-grade detergents to sanitizer solutions for inactivation of E. coli O157:H7 on Romaine lettuce. Freshly-cut leaves of Romaine lettuce were dip-inoculated to achieve a final cell concentration of 7.8 ± 0.2 log CFU/g, air-dried for 2 h, and stored overnight at 4 °C. Leaves were then washed for 2 min in an experimental short chain fatty acid formulation (SCFA) or in one of the following solutions with or without 0.2% dodecylbenzenesulfonic acid or 0.2% sodium 2-ethyl hexyl sulfate: 1) deionized water; 2) 100 ppm chlorine dioxide; 3) 100 ppm chlorine; and 4) 200 ppm chlorine. Following wash treatment, samples were blended in neutralizing buffer (1:3) and surface plated on the selective media CT-SMAC. The efficacy of wash treatments, with or without the detergents, in inactivating E. coli O157:H7 cells on lettuce leaves were not significantly different. The most effective wash solution was SCFA, which was capable of reducing E. coli O157:H7 populations by more than 5 log CFU/g. The rest of the wash treatments resulted in a population reduction of less than 1 log CFU/g. The effectiveness of SCFA surpasses that of other sanitizer treatments tested in this study and requires further research to optimize treatments to preserve lettuce quality. Conventional detergents did not enhance the efficacy of any of the wash treatments tested during this study.