Salmonella sensitivity to sodium hypochlorite and citric acid in washing water of lettuce residues

Salmonella spp. is one of the main lettuce pathogens and should be inactivated during the disinfection of these vegetables before consumption. In minimally processed vegetable industries, residues of organic matter can prevent the inactivation of this pathogen by disinfectants. The objective of the present work was to evaluate the inactivation of Salmonella isolated from organic lettuce to sodium hypochlorite (25 and 50 ppm) and citric acid (0.5 and 1%) in washing water added with lettuce residues. To do so, a washing water with lettuce residues was elaborated, and Salmonella was added in the order of 10⁶ CFU/ml. Thereafter, each sanitizer was added separately to evaluate its effect on reducing Salmonella counts. After 1, 2, 3, 4, 5, 10, and 15 min of contact with the sanitizers, serial dilutions using neutralizer (0.5% sodium thiosulfate) were performed and each dilution was sown in Xylose-Lysine-Desoxycholate medium. Total aerobic mesophilic counts of wash water with lettuce residues before testing (without Salmonella) and after 15 min of exposure to each sanitizer (with Salmonella) were also performed. In addition, the free chlorine still present in the samples after the contact of sodium hypochlorite with lettuce residues for 15 min. The results demonstrated that 50 and 25 ppm sodium hypochlorite could reduce 6 log CFU/ml of Salmonella in 1 and 3 min of contact, respectively, while 0.5 and 1% citric acid was able to reduce 1.26 and 1.74 log CFU/ml respectively from the same microorganism within 15 min of contact. The total aerobic mesophilic counts of the wash water before being tested were, on average, 1.5 log CFU/ml. After addition of Salmonella, with 15 min of contact with the sanitizer, the results of total counts showed the same magnitude as the Salmonella counts. Organic matter may have reacted with the free chlorine present, reducing chlorine concentrations, since values of 30.4 ppm were observed when the initial concentration should be 50 and 17.1 ppm when the initial concentration should be 25 ppm. Based on the results, sodium hypochlorite demonstrated a greater microbial reduction capacity in wash water with lettuce residues, indicating that it is more appropriate to avoid cross-contamination between batches during sanitation of lettuce in washing tanks.     

A pilot plant scale evaluation of a new process aid for enhancing chlorine efficacy against pathogen survival and cross-contamination during produce wash

Developing food safety intervention technology that can be readily adopted by the industry often requires test conditions that match as closely as possible to those of commercial food processing operations; yet biosafety risks inherent in pathogen studies constrain most experiments to laboratory settings. In this study, we report the first semi-commercial pilot-scale evaluation of a new process aid, T128, for its impact on enhancing the antimicrobial efficacy of chlorinated wash water against pathogen survival and cross-contamination. A non-pathogenic, BSL-1, strain of Escherichia coli O157:H7 was inoculated onto freshly harvested baby spinach leaves and washed with large amounts of freshly cut un-inoculated iceberg lettuce shreds in wash water with free chlorine periodically replenished, in the presence or absence of T128. Changes in water quality and pathogen survival and cross-contamination were monitored at every 2min intervals for up to 36min for each treatment during the wash operation. Results indicated that the use of T128 did not significantly (P>0.05) influence the rate of wash water deterioration, nor the pathogen populations remaining on the inoculated spinach leaves. However, in the absence of T128 (control), survival of E. coli O157:H7 in wash water and cross-contamination of un-inoculated lettuce frequently occurred when free chlorine in solution dropped below 1mg/l during the wash process. In contrast, the use of T128 significantly reduced the occurrence of E. coli O157:H7 surviving in wash water and of cross-contamination to un-inoculated shredded iceberg lettuce under the same operational conditions, suggesting that the application of T128 in a chlorine-based fresh produce sanitization system could increase the safety margin of process control on fresh-cut operations.     

Efficacy of a novel sanitizer composed of lactic acid and peroxyacetic acid against single strains of nonpathogenic Escherichia coli K-12, Listeria innocua, and Lactobacillus plantarum in aqueous solution and on surfaces of romaine lettuce and spinach

A novel sanitizer composed of lactic acid and peroxyacetic acid (LA-PAA) was developed as an alternative to chlorinated water (CW) for fresh produce processing. Single strains of Lactobacillus plantarum, nonpathogenic Escherichia coli K-12, and Listeria innocua were used to demonstrate the microbial efficacy of LA-PAA. LA-PAA achieved a >7.8-log reduction of L. innocua and L. plantarum suspended in water at 4°C for 20 s, and LA, PAA, and CW achieved reductions of 0.4, 4.8, and 2.7 log, respectively. LA-PAA, when compared with LA, PAA, and CW, enhanced the reduction of L. innocua attached to romaine leaves by >2.2 log, and improved the removal of E. coli attached to spinach leaves by >2.4 log. The exponential improvement in the microbial efficacy of LA-PAA showed synergism between LA and PAA. LA-PAA microbial efficacy was inversely proportional to pH value and directly correlated with residence time and concentration. Despite an improvement in microbial reduction through the addition of surfactant to LA-PAA, the usage of surfactant in washing fresh produce was impeded by excessive foaming during actual processing. Effects of organic matter on the performance of LA-PAA were minimal. External sensory evaluations showed that LA-PAA had no negative effects on the quality of lettuce and tender leaves. Temperature-abuse studies demonstrated that LA-PAA reduced decay by ～50% when compared with CW. Overall, these results support the premise that LA-PAA has significant potential to be an alternative to CW for fresh produce processing.     

Efficacy of chlorine and a peroxyacetic acid sanitizer in killing Listeria monocytogenes on iceberg and Romaine lettuce using simulated commercial processing conditions

The efficacy of chlorine (100 microg/ml) and a peroxyacetic acid sanitizer (80 microg/ml; Tsunami 100) in killing Listeria monocytogenes inoculated at populations of 1 to 2, 2 to 3, and 4 to 5 log CFU/g of iceberg lettuce pieces, shredded iceberg lettuce, and Romaine lettuce pieces was determined by treatment conditions simulating those used by a commercial fresh-cut lettuce processor. The lettuce/treatment solution ratio was 1:100 (wt/vol), treatment temperature was 4 degrees C, and total treatment time was 30 s. Compared with washing in water, treatment of iceberg lettuce pieces containing all levels of inoculum and shredded iceberg lettuce containing 2 to 3 or 4 to 5 log CFU/g with chlorine or Tsunami resulted in significant reductions (P < or = 0.05) of pathogen populations. Populations recovered from Romaine lettuce pieces treated with chlorine or Tsunami were not significantly different from populations recovered from pieces washed with water, regardless of the inoculum level. Within lettuce type and inoculum level, in no instance was the number of L. monocytogenes recovered from lettuce treated with chlorine or Tsunami significantly different. The rate of decrease in free chlorine concentration in treatment solution as affected by the weight/volume ratio (1:100, 1:10, 2:10, and 4:10) of lettuce and solution was determined. The rate of reduction increased as the ratio decreased. The overall order of magnitude of reduction was shredded iceberg lettuce > iceberg pieces > Romaine pieces. The highest reductions in free chlorine concentration in solutions used to treat shredded lettuce are attributed to the release of tissue juices, which increases the concentration of soluble organic materials available for reaction with chlorine.     

Influence of inoculation method, spot inoculation site, and inoculation size on the efficacy of acidic electrolyzed water against pathogens on lettuce

The influence of bacterial inoculation methods on the efficacy of sanitizers against pathogens was examined. Dip and spot inoculation methods were employed in this study to evaluate the effectiveness of acidic electrolyzed water (AcEW) and chlorinated water (200 ppm free available chlorine) against Escherichia coli O157:H7 and Salmonella spp. Ten pieces of lettuce leaf (5 by 5 cm) were inoculated by each method then immersed in 1.5 liters of AcEW, chlorinated water, or sterile distilled water for 1 min with agitation (150 rpm) at room temperature. The outer (abaxial) and inner (adaxial) surfaces of the lettuce leaf were distinguished in the spot inoculation. Initial inoculated pathogen population was in the range 7.3 to 7.8 log CFU/g. Treatment with AcEW and chlorinated water resulted in a 1 log CFU/g or less reduction of E. coli O157:H7 and Salmonella populations inoculated with the dip method. Spot inoculation of the inner surface of the lettuce leaf with AcEW and chlorinated water reduced the number of E. coli O157:H7 and Salmonella by approximately 2.7 and 2.5 log CFU/g, respectively. Spot inoculation of the outer surface of the lettuce leaf with both sanitizers resulted in approximately 4.6 and 4.4 log CFU/g reductions of E. coli O157:H7 and Salmonella, respectively. The influence of inoculation population size was also examined. Each sanitizer could not completely eliminate the pathogens when E. coli O157:H7 and Salmonella cells inoculated on the lettuce were of low population size (10(3) to 10(4) CFU/g), regardless of the inoculation technique.     

Chlorine stabilizer T-128 enhances efficacy of chlorine against cross-contamination by E. coli O157:H7 and Salmonella in fresh-cut lettuce processing

During fresh-cut produce processing, organic materials released from cut tissues can rapidly react with free chlorine in the wash solution, leading to the potential survival of foodborne bacterial pathogens, and cross-contamination when the free chlorine is depleted. A reported chlorine stabilizer, T-128, has been developed to address this problem. In this study, we evaluated the ability of T-128 to stabilize free chlorine in wash solutions in the presence of high organic loads generated by the addition of lettuce extract or soil. Under conditions used in this study, T-128 significantly (P<0.001) decreased the rate of free chlorine depletion at the presence of soil. T-128 also slightly decreased the rate of free chlorine depletion caused by the addition of lettuce extract in wash solution. Application of T-128 significantly reduced the survival of bacterial pathogens in wash solutions with high organic loads and significantly reduced the potential of cross-contamination, when contaminated and uncontaminated produce were washed together. However, T-128 did not enhance the efficacy of chlorinated wash solutions for microbial reduction on contaminated iceberg lettuce. Evaluation of several produce quality parameters, including overall visual appearance, package headspace O2 and CO2 composition, and lettuce electrolyte leakage, during 15 d of storage indicated that iceberg lettuce quality and shelf life were not negatively impacted by washing fresh-cut lettuce in chlorine solutions containing 0.1% T-128. PRACTICAL APPLICATION: Reported chlorine stabilizer is shown to enhance chlorine efficacy against potential bacterial cross-contamination in the presence of high organic loads without compromising product quality and shelf life.     

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.     

A comparison of different chemical sanitizers for inactivating Escherichia coli O157:H7 and Listeria monocytogenes in solution and on apples, lettuce, strawberries, and cantaloupe

Ozone (3 ppm), chlorine dioxide (3 and 5 ppm), chlorinated trisodium phosphate (100- and 200-ppm chlorine), and peroxyacetic acid (80 ppm) were assessed for reduction of Escherichia coli O157:H7 and Listeria monocytogenes in an aqueous model system and on inoculated produce. Initially, sanitizer solutions were inoculated to contain approximately 10(6) CFU/ml of either pathogen, after which aliquots were removed at 15-s intervals over a period of 5 min and approximately plated to determine log reduction times. Produce was dip inoculated to contain approximately 10(6) E. coli O157:H7 or L. monocytogenes CFU/g, held overnight, submerged in each sanitizer solution for up to 5 min, and then examined for survivors. In the model system study, both pathogens decreased > 5 log following 2 to 5 min of exposure, with ozone being most effective (15 s), followed by chlorine dioxide (19 to 21 s), chlorinated trisodium phosphate (25 to 27 s), and peroxyacetic acid (70 to 75 s). On produce, ozone and chlorine dioxide (5 ppm) were most effective, reducing populations approximately 5.6 log, with chlorine dioxide (3 ppm) and chlorinated trisodium phosphate (200 ppm chlorine) resulting in maximum reductions of approximately 4.9 log. Peroxyacetic acid was the least effective sanitizer (approximately 4.4-log reductions). After treatment, produce samples were stored at 4 degrees C for 9 days and quantitatively examined for E. coli O157:H7, L. monocytogenes, mesophilic aerobic bacteria, yeasts, and molds. Populations of both pathogens remained relatively unchanged, whereas numbers of mesophilic bacteria increased 2 to 3 log during storage. Final mold and yeast populations were significantly higher than initial counts for chlorine dioxide- and ozone-treated produce. Using the nonextended triangle test, whole apples exposed to chlorinated trisodium phosphate (200 ppm chlorine) and shredded lettuce exposed to peroxyacetic acid were statistically different from the other treated samples.     

Pathogen Reduction and Quality of Lettuce Treated with Electrolyzed Oxidizing and Acidified Chlorinated Water

ABSTRACT: The efficacy of electrolyzed oxidizing (EO) and acidified chlorinated water (45 ppm residual chlorine) was evaluated in killing Escherichia coli O157:H7 and Listeria monocytogenes on lettuce. After surface inoculation, each leaf was immersed in 1.5 L of EO or acidified chlorinated water for 1 or 3 min at 22 °C. Compared to a water wash only, the EO water washes significantly decreased mean populations of E. coli O157:H7 and L. monocytogenes by 2.41 and 2.65 log10 CFU per lettuce leaf for 3 min treatments, respectively (p < 0.05). However, the difference between the bactericidal activity of EO and acidified chlorinated waters was not significant (p > 0.05). Change in the quality of lettuce subjected to the different wash treatments was not significant at the end of 2 wk of storage.     

Influence of variations in methodology on populations of Listeria monocytogenes recovered from lettuce treated with sanitizers

The elimination of Listeria monocytogenes inoculated onto a piece of cut iceberg lettuce (3.8 by 3.8 cm) by treatment with chlorinated water (200 micrograms/ml free chlorine) and a 0.5% (wt/vol) solution of FIT Professional Line Antibacterial Cleaner (FIT) was investigated. The efficacy of the two sanitizers was not influenced by the composition of the medium used to culture the L. monocytogenes used in the inocula, the number of strains in the inoculum, or the recovery medium used to enumerate the pathogen on lettuce after treatment. Drying inoculum on lettuce for 45 min at 37 degrees C caused more cells to die or not be retrieved compared with drying inoculum for 30 min at 25 degrees C. However, the percentage of cells in the inoculum recovered from lettuce treated with chlorine or FIT was not significantly different, regardless of the drying method. Stomaching, homogenizing, or stomaching followed by homogenizing lettuce treated with sanitizers resulted in recovery of similar numbers of L. monocytogenes, indicating that stomaching and homogenizing are equivalent in extracting cells; the sequential use of both processing methods did not substantially increase the efficiency of recovery. Washing lettuce with water or treating lettuce with 200 micrograms/ml chlorine or FIT resulted in decreases in populations of 0.60, 1.76, and 1.51 log CFU per lettuce piece, respectively, regardless of variations in test parameters. Reductions caused by sanitizers were significantly greater (alpha = 0.05) than that observed for water but not significantly different from each other. It is concluded that evaluation of sanitizers for their efficacy in killing L. monocytogenes on lettuce can be determined by spot inoculating 50 microliters of a five-strain mixture of cells from 24-h cultures suspended in 5% horse serum albumen, followed by drying the inoculum for 45 min at 37 degrees C, treatment by submerging in 50 ml of sanitizer for 5 min, stomaching samples in 50 ml of Dey-Engley neutralizing broth for 2 min, and enumerating survivors on modified Oxford medium.     

Efficacy of sanitizers to inactivate Escherichia coli O157:H7 on fresh-cut carrot shreds under simulated process water conditions

Chlorine is widely used as a sanitizer to maintain the microbial quality and safety of fresh-cut produce; however, chlorine treatment lacks efficacy on pathogen reduction, especially when the fresh-cut processing water contains heavy organic loads. A more efficacious sanitizer that can tolerate the commercial processing conditions is needed to maintain microbial safety of fresh-cut produce. This study evaluated the efficacy of Escherichia coli O157:H7 reduction on fresh-cut carrots using new and traditional sanitizers with tap water and fresh-cut processing water scenarios. Fresh-cut carrot shreds inoculated with E. coli O157:H7 were washed in sanitizer solutions including 200 ppm chlorine, citric acid-based sanitizer (Pro-San), 80 ppm peroxyacetic acid-based sanitizer (Tsunami 100), and 1,000 ppm acidified sodium chlorite (SANOVA) prepared in fresh tap water or simulated processing water with a chemical oxygen demand level of approximately 3,500 mg/liter. Samples were packaged and stored at 5 degrees C. Microbial analyses performed at days 0, 7, and 14 indicate that the organic load in the process water significantly affected the efficacy of chlorine on pathogen removal and was especially evident on samples tested during storage. Acidified sodium chlorite provided a strong pathogen reduction even under process water conditions with up to a 5.25-log reduction when compared with the no-wash control. E. coli O157:H7 was not recovered on acidified sodium chlorite-treated samples during the entire 14 days of storage, even following an enrichment step. These results suggest that acidified sodium chlorite holds considerable promise as an alternative sanitizer of fresh-cut produce.     

Survival of Enterobacter sakazakii on fresh produce as affected by temperature, and effectiveness of sanitizers for its elimination

A study was done to determine the survival characteristics of Enterobacter sakazakii on the surface of apples, cantaloupes, strawberries, lettuce, and tomatoes stored at 4, 12, and 25 degrees C for 8-28 days. Populations significantly decreased (por=50 microg/ml, were equivalent in killing E. sakazakii on apples. Populations of E. sakazakii on apples treated with 10 microg/ml chlorine dioxide for 1 or 5 min were significantly reduced (por=4.00 log CFU/apple. Reductions of >or=3.70 log CFU/tomato were achieved by treatment with 10 microg/ml chlorine or chlorine dioxide or 40 microg/ml Tsunami 200 for 5 min. Reductions in populations of E. sakazakii on lettuce treated with chlorine at 10, 50, and 100 microg/ml for 1 min ranged from 1.61 to 2.50 log CFU/sample (26+/-4 g), compared to populations remaining on lettuce washed with water. Chlorine was less effective in killing E. sakazakii on lettuce than on apples or tomatoes. Treatment of lettuce with Tsunami 200 (40 and 80 microg/ml) for 5 min caused a reduction of >or=5.31 log CFU/sample. Results provide insights to predicting survival characteristics of E. sakazakii on produce and the efficacy of sanitizers in killing the bacterium.     

Determination of free chlorine concentrations needed to prevent Escherichia coli O157:H7 cross-contamination during fresh-cut produce wash

This study was conducted to investigate the effect of free chlorine concentrations in wash water on Escherichia coli O157:H7 reduction, survival, and transference during washing of fresh-cut lettuce. The effectiveness of rewashing for inactivation of E. coli O157:H7 on newly cross-contaminated produce previously washed with solutions containing an insufficient amount of chlorine also was assessed. Results indicate that solutions containing a minimum of 0.5 mg/liter free chlorine were effective for inactivating E. coli O157:H7 in suspension to below the detection level. However, the presence of 1 mg/liter free chlorine in the wash solution before washing was insufficient to prevent E. coli O157:H7 survival and transfer during washing because the introduction of cut lettuce to the wash system quickly depleted the free chlorine. Although no E. coli O157:H7 was detected in the wash solution containing 5 mg/liter free chlorine before washing a mix of inoculated and uninoculated lettuce, low numbers of E. coli O157:H7 cells were detected on uninoculated lettuce in four of the seven experimental trials. When the prewash free chlorine concentration was increased to 10 mg/liter or greater, no E. coli O157:H7 transfer was detected. Furthermore, although rewashing newly cross-contaminated lettuce in 50 mg/liter free chlorine for 30 s significantly reduced (P = 0.002) the E. coli O157:H7 populations, it failed to eliminate E. coli O157:H7 on lettuce. This finding suggests that rewashing is not an effective way to correct for process failure, and maintaining a sufficient free chlorine concentration in the wash solution is critical for preventing pathogen cross-contamination.     

Cross-contamination of fresh-cut lettuce after a short-term exposure during pre-washing cannot be controlled after subsequent washing with chlorine dioxide or sodium hypochlorite

Chlorine dioxide (ClO₂) has been postulated as an alternative to sodium hypochlorite (NaClO) for fresh-cut produce sanitization to avoid risks associated with chlorination by-products. Experiments were performed to determine the prevention of cross-contamination of fresh-cut lettuce by Escherichia coli using chlorine dioxide (3 mg/L) or sodium hypochlorite (100 mg/L) as sanitation agents. The efficacy of these sanitation solutions was evaluated simulating as much as possible the conditions of a fresh-cut processing line. Thus, to evaluate the potential risk of cross-contamination during pre-washing, inoculated fresh-cut lettuce was pre-washed and after that non-inoculated lettuce was then pre-washed in the same water. After this pre-washing, non-inoculated lettuce was cross-contaminated, changing from 0 to 3.4 log units of E. coli cells. During washing with sanitizers, none of the tested sanitation agents significantly reduced E. coli counts in both inoculated and cross-contaminated lettuce. These results suggest that when cross-contamination occurs, even if the event is recent, subsequent sanitation steps are inefficient for inactivating E. coli cells on the vegetable tissue. However, chlorine dioxide and sodium hypochlorite solutions were able to inactivate most E. coli cells that passed from inoculated product to wash water. Therefore, they might be able to avoid cross-contamination between clean and contaminated product during the washing step. Scanning electron microscopy micrographs indicated that bacterial cells were mainly located in clusters or tissue stomata where they might be protected, which explains the low efficacy of sodium hypochlorite and chlorine dioxide solutions observed in this study.     

Sanitation and design of lettuce coring knives for minimizing Escherichia coli O157:H7 contamination

This study was undertaken to examine the effect of ultrasound in combination with chlorine on the reduction of Escherichia coli O157:H7 populations on lettuce coring knives. Two new coring devices designed to mitigate pathogen attachment were fabricated and evaluated. The coring rings of the knives were dip inoculated with soil slurry containing 10? E. coli cells and treated with chlorinated water with and without ultrasonication for 30, 60, and 120 s. The rough welding joints on currently used in-field lettuce coring knives provided a site conducive to bacterial attachment and resistant to cell removal during sanitation treatment. The two modified coring knives harbored significantly fewer E. coli cells than did the currently used commercial model, and the efficacy of the disinfection treatment was high (P < 0.05). Ultrasound treatment reduced the E. coli O157:H7 counts to below the detection limit of 1.10 log CFU/cm2 at both the coring ring blade and welding joint within 30 s in 1 ppm of chlorinated water. The redesigned coring knives and an ultrasound plus chlorine combination treatment may provide practical options for minimizing the microbial safety hazards of lettuce processed by core-in-field operations.     

Antimicrobial effect of electrolyzed water for inactivating Campylobacter jejuni during poultry washing

The effectiveness of electrolyzed (EO) water for killing Campylobacter jejuni on poultry was evaluated. Complete inactivation of C. jejuni in pure culture occurred within 10 s after exposure to EO or chlorinated water, both of which contained 50 mg/l of residual chlorine. A strong bactericidal activity was also observed on the diluted EO water (containing 25 mg/l of residual chlorine) and the mean population of C. jejuni was reduced to less than 10 CFU/ml (detected only by enrichment for 48 h) after 10-s treatment. The diluted chlorine water (25 mg/l residual chlorine) was less effective than the diluted EO water for inactivation of C. jejuni. EO water was further evaluated for its effectiveness in reducing C. jejuni on chicken during washing. EO water treatment was equally effective as chlorinated water and both achieved reduction of C. jejuni by about 3 log10 CFU/g on chicken, whereas deionized water (control) treatment resulted in only 1 log10 CFU/g reduction. No viable cells of C. jejuni were recovered in EO and chlorinated water after washing treatment, whereas high populations of C. jejuni (4 log10 CFU/ml) were recovered in the wash solution after the control treatment. Our study demonstrated that EO water was very effective not only in reducing the populations of C. jejuni on chicken, but also could prevent cross-contamination of processing environments.     

Effect of warm,chlorinated water on the microbial flora of shredded iceberg lettuce

An optimization technique was used to determine suitable times of exposure and temperatures for maximum destruction of microorganisms in shredded iceberg lettuce dipped in warm, chlorinated (100 μg/ml) water in a model system. Shredded lettuce washed for 3 min at 47°C and packaged in high oxygen transmission rate (6000–8000 cc m² 24 h) bags maintained an acceptable appearance after 7 days in storage at 1±1°C. Initial microbial loads were reduced by approximately 3 log cfu g⁻¹ in lettuce washed in chlorinated water at 47°C, and 1 log cfu g⁻¹ at 4°C. Pilot plant scale studies confirmed observations in the model system. The microbial flora of shredded lettuce packaged in the raw state and after a 3 min wash in 100 g ml⁻¹ chlorine at 4 or 47°C was dominated by psychrotrophic bacteria, particularly species of Pseudomonas, throughout the storage period. Numbers of these microorganisms were significantly lower than in raw lettuce or lettuce washed at 4°C until the 15th day in storage. Gas composition inside the bags was not affected by the washes.     

Ultrasound decontamination of minimally processed fruits and vegetables

The effectiveness of power ultrasound for the microbial decontamination of minimally processed fruits and vegetables was studied. Reductions in Salmonella typhimurium attached to iceberg lettuce obtained by cleaning with water, chlorinated water, ultrasound with water and ultrasound with chlorinated water were 0.7, 1.7, 1.5 and 2.7 logs, respectively, for small-scale (2 L) trials. The cleaning action of cavitation appears to remove cells attached to the surface of fresh produce, rendering the pathogens more susceptible to the sanitizer. For large-scale (40 L) trials, the addition of chlorine to water in the tank gave a systematic difference in Escherichia coli decontamination efficiency. However, the frequency of ultrasound treatment (25, 32–40, 62–70 kHz) had no significant effect on decontamination efficiency ( P  > 0.69). With the potentially high capital expenditure together with the expensive process of optimization and water treatment, it is unlikely that the fresh produce industry would be willing to take up this technology. Furthermore, the additional one log reduction achieved by applying ultrasound to a chlorinated water wash does not completely eliminate the risk of pathogens on fresh produce.     

Bactericidal effects of lettuce after subsequent washing with hypobromous acid and sodium hypochlorite

Sodium hypochlorite (NaClO) is widely used as a sanitizer to maintain microbial safety of lettuce; however, NaClO treatment is insufficient on pathogen reduction due to its high instability for an extended storage time. Hypobromous acid (HOBr) as another sanitizer, relatively stable in processing conditions, was applied to lettuce. The numbers of total aerobic bacteria, Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus were evaluated on lettuce treated with NaClO and HOBr. The population of 4 selected bacteria on lettuce reduced more than 2.5 log CFU/g. Bactericidal effect maintained for 10 days in combined treatment with 100 mg/L NaClO followed by 200 mg/L HOBr, while the population of the bacteria without HOBr treatment increased. Combined treatment on lettuce can give synergistic effect on reducing microbial population. Residual bromine was not detected on the lettuce treated with HOBr after washed with water for 30 s, indicating that bromine was effectively washed away.     

PREDICTIVE MODELING AND GROWTH MODELS OF AEROBIC MESOPHILIC BACTERIA ON FRESH-CUT LETTUCE BY HYPOCHLORITE-WASHING

Response surface methodology was used to evaluate the effect of three selected factors (chlorine concentration, washing time and water‐to‐lettuce ratio) on reducing aerobic mesophilic bacteria on fresh‐cut lettuce. According to statistical analysis, the model established was effective in predicting the reduction of aerobic mesophilic bacteria on fresh‐cut lettuce by washing with chlorine. In addition, best‐fit Gompertz‐modified models were described to evaluate aerobic mesophilic bacteria growth on fresh‐cut lettuce during storage at 0, 4 and 25C, respectively. The final load of aerobic mesophilic bacteria and shelf life of fresh‐cut lettuce could be predicted in various storing temperatures with the growth models.