Chlorine dioxide gas from an aqueous solution: reduction of Salmonella in wounds on tomato fruit and movement to sinks in a treatment chamber.

Chlorine dioxide (ClO2) off-gassed from an aqueous solution and reacted incrementally with potassium iodide solutions (sinks). After 30 min, 45% of the initial dose was detected as chlorite ion in the sink, whereas 35% of the initial dose was still in the source. Aqueous solutions of ClO2 can be used as a source of ClO2 gas in various laboratory experiments involving treatment of fruits or vegetables. Movement from source to sink is continuous, which precludes the development of large headspace concentrations and the need for a tight chamber seal. When the source solution has dissipated, the chamber can be opened safely as there is little free ClO2 remaining in the headspace. In tests with whole, wound-inoculated tomato fruit, at both green and pink stages of ripeness, the control of Salmonella enterica serotype Typhimurium in wounds varied with the weight of gas used. The number of viable cells of Typhimurium recovered was reduced by > 5 log units when > or = 0.5 mg of ClO2 was applied to three pieces of fruit during a 2-h treatment.

Applications of Chlorine Dioxide Gas for Control of Bacterial Soft Rot in Tomatoes.

Chlorine dioxide (ClO2) gas was generated from a mixture of sodium chlorite and ferric chloride plus water (impregnated into zeolite) in a Tyvek sachet over a 2- or 24-h period. The gas was distributed by a fan over wound-inoculated tomato fruit (Lycopersicon esculentum) enclosed in a sealed aluminum pressure cooker. Within 24 h of inoculation with 6 log10 CFU of Erwinia carotovora subsp. carotovora per wound and storage at 22 to 24°C, bacterial soft rot was observed on >80% of the nontreated wounds (10 wounds/fruit and 4 or 6 fruit/treatment). By contrast, wounds that had been exposed to an atmosphere containing up to 99 mg ClO2 during a 2- or 24-h period remained firm and dry with no evidence of bacterial activity or soft rot. After 72 h of incubation, wounds exposed to 88 mg ClO2 produced over 24 h or 99 mg ClO2 produced over 2 h were free of decay, whereas bacterial soft rot was observed in ca. 12% and less than 5% of wounds treated with 0.75 mg or 7.5 mg, respectively, for either 2 or 24 h. Wounds that had not been inoculated remained free of bacterial soft rot throughout the entire storage period. Wounds exposed to the highest doses of ClO2, 88 mg/24 h or 99 mg/2 h, became bleached and sunken. Additionally, the stem scars on these fruit became cracked, sunken, and bleached. The intact cuticle was not visibly affected, and there was no observed change in overall fruit color. ClO2 gas may be effective for controlling postharvest decays of fruit that have been inoculated prior to or during harvest.

Reduction of poliovirus 1, bacteriophages, Salmonella montevideo, and Escherichia coli O157:H7 on strawberries by physical and disinfectant washes.

The efficacy levels of different physical and chemical washing treatments in the reduction of viral and bacterial pathogens from inoculated strawberries were evaluated. Escherichia coli O157:H7, Salmonella Montevideo, poliovirus 1, and the bacteriophages PRD1, phiX174, and MS2 were used as model and surrogate organisms. Chemicals readily available to producers and/or consumers were evaluated as antimicrobial additives for the production of washes. The gentle agitation of contaminated strawberries in water for 2 min led to reductions in microbial populations ranging from 41 to 79% and from 62 to 90% at water temperatures of 22 and 43 degrees C, respectively. Significant reductions (> 98%) in numbers of bacteria and viruses were obtained with sodium hypochlorite (50 to 300 ppm of free chlorine), Oxine or Carnebon (200 ppm of product generating "stabilized chlorine dioxide"), Tsunami (100 ppm of peroxyacetic acid), and Alcide (100 or 200 ppm of acidified sodium chlorite) washes. Overall, 200 ppm of acidified sodium chlorite produced the greatest reductions of microorganisms. Hydrogen peroxide (0.5%) was slightly less effective than free chlorine in a strawberry wash and caused slight fruit discoloration. Cetylpyridinium chloride (0.1%) was effective in the reduction of bacterial species, while trisodium phosphate (1%) was effective against viruses. The consumer-oriented produce wash Fit was very effective (> 99%) in reducing the numbers of bacteria but not in reducing the numbers of viruses. Another wash, Healthy Harvest, was significantly less effective than Fit in reducing bacterial pathogens but more effective for viruses. The performance of automatic dishwashing detergent was similar to that of Healthy Harvest and significantly better than that of liquid dishwashing detergent. Solutions containing table salt (2% NaCl) or vinegar (10%) reduced the numbers of bacteria by about 90%, whereas only the vinegar wash reduced the numbers of viruses significantly (ca. 95%).

Postharvest Decay Risk Associated with Hydrocooling Tomatoes.

Tomatoes (breaker stage) hydrocooled with a cell suspension of Erwinia carotovora subsp. carotovora containing 50 to 200 mg of free chlorine per liter (ppm) (10°C, pH 7) remained decay free during a 10-day storage at 20°C. Sporadic disease appeared during storage of tomatoes similarly cooled with chlorinated water containing spores of Rhizopus stolonifer. In contrast, when chlorine was omitted from the pathogen suspensions, 50 to 100% of the fruit became diseased. A laboratory-scale shower hydrocooler reduced fruit temperatures from 35 to 15°C within 13.3 min, whereas a flume cooler produced the same temperature reduction in 10.5 min. In both systems, tomatoes increased in weight during cooling, evidence for water uptake. Larger weight increases occurred among tomatoes cooled in the shower than in the flume. An upward instead of downward orientation of stem scars under the shower streams led to significantly larger weight increases, presumably because pores in the stem scar were continuously flooded with water. Tomatoes intermittently submerged in cold water (10 2-min immersions followed by 30-s pauses) absorbed significantly less water than those continuously submerged for 20 min. Hydrocooling appears to be a viable method for rapid cooling of tomatoes. Technical refinements in the hydrocooling process that prevent continuous coverage of fruit surfaces by water should reduce water uptake and the associated risk of pathogen internalization. Maintenance of free chlorine at up to 200 ppm in the cooling water and prevention of direct water pressure on fruit should minimize decay risks. No evidence of phytotoxicity was observed among fruit infiltrated with 200 ppm of chlorine. These tomatoes ripened similarly to those that were not cooled or were cooled in tap water.

Factors that affect proliferation of Salmonella in tomatoes post-harvest: the roles of seasonal effects, irrigation regime, crop and pathogen genotype.

MAIN OBJECTIVES: Fresh fruits and vegetables become increasingly recognized as vehicles of human salmonellosis. Physiological, ecological, and environmental factors are all thought to contribute to the ability of Salmonella to colonize fruits and vegetables pre- and post-harvest. The goal of this study was to test how irrigation levels, fruit water congestion, crop and pathogen genotypes affect the ability of Salmonella to multiply in tomatoes post-harvest. EXPERIMENTAL DESIGN: Fruits from three tomato varieties, grown over three production seasons in two Florida locations, were infected with seven strains of Salmonella and their ability to multiply post-harvest in field-grown tomatoes was tested. The field experiments were set up as a two-factor factorial split plot experiment, with the whole-plot treatments arranged in a randomized complete-block design. The irrigation treatment (at three levels) was the whole-plot factor, and the split-plot factor was tomato variety, with three levels. The significance of the main, two-way, and three-way interaction effects was tested using the (type III) F-tests for fixed effects. Mean separation for each significant fixed effect in the model was performed using Tukey's multiple comparison testing procedure. MOST IMPORTANT DISCOVERIES AND SIGNIFICANCE: The irrigation regime per se did not affect susceptibility of the crop to post-harvest proliferation of Salmonella. However, Salmonella grew significantly better in water-congested tissues of green tomatoes. Tomato maturity and genotype, Salmonella genotype, and inter-seasonal differences were the strongest factors affecting proliferation. Red ripe tomatoes were significantly and consistently more conducive to proliferation of Salmonella. Tomatoes harvested in the driest, sunniest season were the most conducive to post-harvest proliferation of the pathogen. Statistically significant interactions between production conditions affected post-harvest susceptibility of the crop to the pathogen. UV irradiation of tomatoes post-harvest promoted Salmonella growth.

Untangling metabolic and communication networks: interactions of enterics with phytobacteria and their implications in produce safety.

Recent outbreaks of vegetable-borne gastrointestinal illnesses across the globe demonstrate that human enteric pathogens can contaminate produce at any stage of production. Interactions of enterics with native plant-associated microbiota influence the microbiological safety of produce by affecting the attachment, persistence and proliferation of human pathogens on plants. Supermarket surveys have revealed that bacteria, but not fungi or mechanical damage, promote the growth of Salmonella enterica on produce. Field and laboratory studies have indicated that some plant pathogenic bacteria and fungi facilitate the entry and internalization of human pathogens in plants. Conversely, some phytobacteria, including those involved in biocontrol of plant diseases, significantly inhibit attachment and plant colonization by non-typhoidal Salmonella and enterovirulent Escherichia coli by producing antibiotics or competing for nutrients in the phyllosphere. In this review, we attempt to elucidate the mechanisms of interactions between human enteric pathogens and plant-associated microbiota, and describe how these interactions affect produce safety.

Internal colonization of Salmonella enterica serovar Typhimurium in tomato plants.

Several Salmonella enterica outbreaks have been traced back to contaminated tomatoes. In this study, the internalization of S. enterica Typhimurium via tomato leaves was investigated as affected by surfactants and bacterial rdar morphotype, which was reported to be important for the environmental persistence and attachment of Salmonella to plants. Surfactants, especially Silwet L-77, promoted ingress and survival of S. enterica Typhimurium in tomato leaves. In each of two experiments, 84 tomato plants were inoculated two to four times before fruiting with GFP-labeled S. enterica Typhimurium strain MAE110 (with rdar morphotype) or MAE119 (without rdar). For each inoculation, single leaflets were dipped in 10(9) CFU/ml Salmonella suspension with Silwet L-77. Inoculated and adjacent leaflets were tested for Salmonella survival for 3 weeks after each inoculation. The surface and pulp of ripe fruits produced on these plants were also examined for Salmonella. Populations of both Salmonella strains in inoculated leaflets decreased during 2 weeks after inoculation but remained unchanged (at about 10(4) CFU/g) in week 3. Populations of MAE110 were significantly higher (P<0.05) than those of MAE119 from day 3 after inoculation. In the first year, nine fruits collected from one of the 42 MAE119 inoculated plants were positive for S. enterica Typhimurium. In the second year, Salmonella was detected in adjacent non-inoculated leaves of eight tomato plants (five inoculated with strain MAE110). The pulp of 12 fruits from two plants inoculated with MAE110 was Salmonella positive (about 10(6) CFU/g). Internalization was confirmed by fluorescence and confocal laser microscopy. For the first time, convincing evidence is presented that S. enterica can move inside tomato plants grown in natural field soil and colonize fruits at high levels without inducing any symptoms, except for a slight reduction in plant growth.