Efficacy of Chlorine Dioxide Gas Against Hepatitis A Virus on Blueberries, Blackberries, Raspberries, and Strawberries.

Seeking a means of sanitizing berries, the effectiveness of steady state levels of gaseous chlorine dioxide (ClO2) against hepatitis A virus (HAV) on laboratory-contaminated berries was determined. The generated ClO2 was maintained with 1 or 2 mg/l air inside a 269-l glove box to treat 50 g batches of blueberries, raspberries, and blackberries, and 100 g batches of strawberries that were immersion coated with HAV. Normalized data for ClO2 (ppm-h/g product) is reported as a function of ClO2 concentration, treatment time, and weight of treated product. Treatments of ClO2 ranging from 1.00 to 6.27 ppm-h/g berry were evaluated. When compared to untreated HAV-contaminated berries, log reductions of HAV were > 2.1 for all berry types and conditions tested indicating the gaseous ClO2 was effective. The average log reduction with strawberries, raspberries, blueberries and blackberries treated with 1.00 ppm-h/g, the lowest ClO2 treatment tested, were 2.44, 2.49, 3.23, and 3.45, respectively. The highest treatment of 6.27 ppm-h/g was applied at two different gas concentrations of 1 mg/l and 2 mg/l. Average log reductions for blueberries and strawberries treated with 6.27 ppm-h/g were 4.34 and 4.42, and 4.03 and 3.51, applied at 1 mg/l and 2 mg/l, respectively. For blackberries and raspberries 3.20 and 3.24, and 3.23 and 3.97 log reductions were observed for 6.27 ppm-h/g treatments applied at 1 mg/l and 2 mg/l, respectively. Results indicate that HAV contamination of berries can be substantially reduced by gaseous ClO2 and offer industry a waterless means of sanitizing berries against HAV.

Evaluation of Chlorine Dioxide Gas against Four Salmonella enterica Serovars Artificially Contaminated on Whole Blueberries.

ABSTRACT: Fresh produce, such as blueberries, continues to be a source of foodborne illness in the United States. Despite new practices and intervention technologies, blueberries and other produce are contaminated with foodborne pathogens, such as Salmonella. The aim of this study was to evaluate the efficacy of chlorine dioxide gas (CDG) against Salmonella enterica serovars Newport, Stanley, Muenchen, and Anatum on artificially contaminated whole fresh blueberries. Blueberries were dip inoculated into a 400-mL bath containing a Salmonella serovar cocktail of either ca. 6 or 9 log CFU/mL. Samples were dried for either 2 or 24 h before treatment with 1.5 or 3 mg of CDG/L of air to a final treatment of 3.55 to 6 ppm/h/g of blueberry. Salmonella cells were recovered by stomaching CDG-treated and nontreated control samples with 0.1% peptone and enumerated on xylose lysine Tergitol 4 agar. CDG treatments achieved up to a 5.63-log CFU/g reduction of the cocktail using 5.5 ppm/h/g, whereas the lowest treatment, 4 ppm/h/g (1.5 mg of CDG/L), was still capable of a 4.45-log CFU/g reduction. Incubation time significantly (P < 0.001) affected CDG efficacy against both inoculation concentrations. Additionally, all serovars responded similarly to CDG treatment when tested individually (P > 0.0691). Finally, the availability of a water reservoir during treatments did not have a significant effect (P = 0.9818) on CDG efficacy in this study. Our results demonstrate that CDG can be an efficacious treatment option for whole blueberry decontamination.

Challenges in Recovering Foodborne Pathogens from Low-Water-Activity Foods.

There are numerous obstacles to the detection of foodborne pathogens in foods that exhibit a low water activity (aw). These obstacles include the presence of antimicrobial compounds, particulates, PCR inhibitors, and fatty matrices. New approaches should be sought to increase the sensitivity of pathogen testing in low-aw foods and to overcome the effects of various inhibitors and antimicrobials. The U.S. Food and Drug Administration and other laboratories are working toward this goal. This review will address these issues while delineating specific inhibitors and antimicrobials that impede testing of low-aw foods. A review of relevant rapid and conventional testing methodologies for Salmonella in low-aw foods will also be discussed.

Evaluation of Steady-State Gaseous Chlorine Dioxide Treatment for the Inactivation of Tulane virus on Berry Fruits.

The effectiveness of steady-state levels of gaseous chlorine dioxide (ClO2) against Tulane virus (TV), a human norovirus surrogate, on berries was determined. The generated ClO2 was maintained at 1 mg/L inside a 269 L glove box to treat two 50 g batches of blueberries, raspberries, and blackberries, and two 100 g batches of strawberries that were immersion coated with TV. The standardized/normalized treatment concentrations of ClO2 ranging from 0.63 to 4.40 ppm-h/g berry were evaluated. When compared to untreated TV contaminated berries, log reductions of TV were in excess of 2.9 log PFU/g for all berry types and conditions tested, indicating that ClO2 was highly effective. In general, the efficacy of all ClO2 treatments on log reductions of TV on all berries was not significantly different (p < 0.05). The average log reduction with strawberries, raspberries, blueberries, and blackberries, treated with the lowest ClO2 concentration, 0.63 ppm-h/g, were 2.98, 3.40, 3.82, and 4.17 log PFU/g, respectively. Overall results suggest that constant levels of ClO2 could be quite effective against foodborne viruses.

Evaluation of a Male-Specific DNA Coliphage Persistence Within Eastern Oysters (Crassostrea virginica).

Male-specific coliphages (MSCs) are currently used to assess the virologic quality of shellfish-growing waters and to assess the impact of sewage release or adverse weather events on bivalve shellfish. Since MSC can have either DNA or RNA genomes, and most research has been performed exclusively on RNA MSCs, persistence of M13, a DNA MSC, was evaluated for its persistence as a function of time and temperature within Eastern oysters (Crassostrea virginica). Oysters were individually exposed to seawater containing a total of 1010 to 1012 pfu of M13 for 24 h at 15 °C followed by maintenance in tanks with as many as 21 oysters in continuously UV-sterilized water for up to 6 weeks at either 7, 15, or 22 °C. Two trials for each temperature were performed combining three shucked oysters per time point which were assayed by tenfold serial dilution in triplicate. Initial contamination levels averaged 106.9 and ranged from 106.0 to 107.0 of M13. For oysters held for 3 weeks, log10 reductions were 1.7, 3.8, and 4.2 log10 at 7, 15, and 22 °C, respectively. Oysters held at 7 and 15 °C for 6 weeks showed average reductions of 3.6 and 5.1 log10, respectively, but still retained infectious M13. In total, this work shows that DNA MSC may decline within shellfish in a manner analogous to RNA MSCs.

Evaluation of Hot Water, Gaseous Chlorine Dioxide, and Chlorine Treatments in Combination with an Edible Coating for Enhancing Safety, Quality, and Shelf Life of Fresh-Cut Cantaloupes.

Fresh-cut cantaloupes have been implicated in numerous foodborne outbreaks of salmonellosis. Commercial aqueous wash treatments are limited in their ability to inactivate Salmonella enterica. Our objective was to evaluate the efficacy of hot water, gaseous chlorine dioxide, and chlorine on enhancing microbial safety and sensory qualities of fresh-cut cantaloupes. Cantaloupes were inoculated with an S. enterica cocktail (serovars Michigan, Mbandaka, and Poona) and treated with chlorine (200 ppm of free chlorine) for 40 min, 5 mg/L gaseous chlorine dioxide for 4.5 h, and hot water (76.1°C) for 3 min. Fresh-cut cantaloupes were prepared from treated whole cantaloupes and divided into two sets; one set of samples was treated with NatureSeal to evaluate its effect on shelf life and sensory quality and the second set (control) was packed without further treatment. Fresh-cut samples were stored at 4°C for up to 21 days. For the sensory quality parameters analyzed (color, water loss, and texture), the samples treated with NatureSeal had significantly better quality ( P < 0.05) than did the control samples. All treatments significantly reduced ( P < 0.05) the pathogen populations on the rind of the whole melons and on the fresh-cut samples prepared from the treated melons. All fresh-cut samples prepared from melons treated with hot water were negative for Salmonella throughout the storage period except for the samples treated with hot water and NatureSeal and evaluated on day 7. The fresh-cut samples prepared from melons treated with chlorine dioxide and chlorine were negative for Salmonella after 21 days of storage. These results provide a framework to producers of fresh-cut cantaloupes for the potential use of hot water as an intervention treatment in combination with NatureSeal for enhancing the microbiological safety and quality of this commodity.

Survival of Salmonella Typhimurium on soybean sprouts following treatments with gaseous chlorine dioxide and biocontrol Pseudomonas bacteria.

Control of Salmonella Typhimurium on sprouts is crucial for food and consumer safety. In this study, natural microflora on soybean seed was assessed and effects of gaseous chlorine dioxide (ClO2) and biocontrol Pseudomonas on the survival of S. Typhimurium on soybean sprouts were evaluated. Sprouts were dip-inoculated with S. Typhimurium prior to the application of the biocontrol (P. chlororaphis and P. fluorescens). After inoculation with S. Typhimurium, the sprouts were treated with ClO2 at 0.4 mg/L for 1 h (90% R.H., 13°C). Pseudomonas strains and Salmonella were recovered on Pseudomonas Agar F (PAF) and xylose lysine tergitol-4 (XLT-4) media, respectively. Pseudomonas strains reduced Salmonella by <1 log colony forming units (CFU)/g of sprouts, whereas S. Typhimurium on soybean sprouts was reduced from 2.55 to 5.35 logs CFU/g by ClO2. Gaseous ClO2 treatment reduced S. Typhimurium by 3.90 (0 h), 4.47 (24 h), and 3.61 log CFU/g (168 h). It was concluded that ClO2 and biocontrol treatment can enhance sprout safety.

Development of Combined Dry Heat and Chlorine Dioxide Gas Treatment with Mechanical Mixing for Inactivation of Salmonella enterica Serovar Montevideo on Mung Bean Seeds.

Foodborne outbreaks have been associated with the consumption of fresh sprouted beans. The sprouting conditions of mung bean seeds provide optimal conditions of temperature and relative humidity for any potential pathogenic contaminant on the seeds to grow. The lack of a kill step postsprouting is a major safety concern. Thus, the use of a kill step on the seeds prior to a sprouting step would enhance the safety of fresh sprouts. The objective of this work was to evaluate the effectiveness of the combined thermal and chlorine dioxide gas (3.5 mg/liter of air) treatment with mechanical mixing (tumbling) to eliminate Salmonella on artificially inoculated mung bean seeds. Although no viable Salmonella was recovered from seeds treated in hot water at 60°C for 2 h, these treated seeds failed to germinate. Dry heat treatments (55, 60, or 70°C) for up to 8 h reduced Salmonella populations in excess of 3 log CFU/g. The use of tumbling, while treating the seeds, resulted in up to 1.6 log CFU/g reduction in Salmonella populations compared with no tumbling. Dry heat treatment at 65°C for 18 h with tumbling resulted in a complete inactivation of Salmonella populations on inoculated seeds with low inoculum levels (2.83 log CFU/g) as compared with high inoculum levels (4.75 log CFU/g). The increased reductions in pathogenic populations on the seeds with the use of tumbling could be attributed to increased uniformity of heat transfer and exposure to chlorine dioxide gas. All treated seeds were capable of germinating, as well as the nontreated controls. These results suggest that this combined treatment would be a viable process for enhancing the safety of fresh sprouts.

Evaluation of chlorine dioxide gas treatment to inactivate Salmonella enterica on mungbean sprouts.

Although freshly sprouted beans and grains are considered to be a source of nutrients, they have been associated with foodborne outbreaks. Sprouts provide good matrices for microbial localization and growth due to optimal conditions of temperature and humidity while sprouting. Also, the lack of a kill step postsprouting is a major safety concern. The objective of this work was to evaluate the effectiveness of chlorine dioxide gas treatment to reduce Salmonella on artificially inoculated mungbean sprouts. The effectiveness of gaseous chlorine dioxide (0.5 mg/liter of air) with or without tumbling (mechanical mixing) was compared with an aqueous chlorine (200 ppm) wash treatment. Tumbling the inoculated sprouts during the chlorine dioxide gas application for 15, 30, and 60 min reduced Salmonella populations by 3.0, 4.0, and 5.5 log CFU/g, respectively, as compared with 3.0, 3.0, and 4.0 log CFU/g reductions obtained without tumbling, respectively. A 2.0 log CFU/g reduction in Salmonella was achieved with an aqueous chlorine wash. The difference in microbial reduction between chlorine dioxide gas versus aqueous chlorine wash points to the important role of surface topography, pore structure, bacterial attachment, and/or biofilm formation on sprouts. These data suggested that chlorine dioxide gas was capable of penetrating and inactivating cells that are attached to inaccessible sites and/or are within biofilms on the sprout surface as compared with an aqueous chlorine wash. Consequently, scanning electron microscopy imaging indicated that chlorine dioxide gas treatment was capable of penetrating and inactivating cells attached to inaccessible sites and within biofilms on the sprout surfaces.

Commercial thermal process for inactivating Salmonella Poona on surfaces of whole fresh cantaloupes.

Outbreaks of salmonellosis by Salmonella Poona and listeriosis by Listeria monocytogenes have been associated with the consumption of cantaloupes. Commercial washing processes for cantaloupes are limited in their ability to inactivate and/or remove this human pathogen. Our objective was to develop a commercial-scale surface pasteurization process for enhancing microbiological safety of cantaloupes. Whole cantaloupes, surface inoculated with Salmonella Poona RM 2350, were stored at 32°C for 24 h prior to processing. Temperature-penetration profiles indicated that the surface temperature of the whole cantaloupe was 26 and 30°C below that of the hot water temperature after 2 min of immersion at 82 and 92°C, respectively. Hot water treatments at 92°C for 60 and 90 s reduced Salmonella Poona populations in excess of 5 log CFU/g of rind. Cantaloupes that were treated and stored at 4°C for 9 days retained their firmness quality and maintained nondetectable levels of Salmonella Poona as compared with the controls. Also, levels of Salmonella Poona on fresh-cut cantaloupe prepared from hot water-treated cantaloupes and stored for 9 days at 4°C were nondetectable as compared with the controls. These results indicate that surface pasteurization at 92°C for 90 s will enhance the microbiological safety of cantaloupes and will extend the shelf life of this commodity as well. The process parameter of 90 s or less falls within the commercial requirements of the whole-cantaloupe processor-packer industry.

Management of microbial food safety in Arab countries.

Microbial food safety remains a major economic and public health concern in Arab countries. Over the past several years, many of these countries have attempted to revise and upgrade food quality control and surveillance programs; however, these systems vary in scope and effectiveness. This review addresses the major reported foodborne outbreaks and multidrug resistance of pathogenic microorganisms isolated from food products. Major foodborne pathogens of concern included Brucella spp., Clostridium botulinum, fecal coliforms, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella, and Staphylococcus aureus. Measures for managing microbial food hazards based on a comprehensive risk analysis also are proposed.

Application of ozonated dry ice (ALIGAL™ Blue Ice) for packaging and transport in the food industry.

Dry ice is used by meat and poultry processors for temperature reduction during processing and for temperature maintenance during transportation. ALIGAL™ Blue Ice (ABI), which combines the antimicrobial effect of ozone (O(3)) along with the high cooling capacity of dry ice, was investigated for its effect on bacterial reduction in air, in liquid, and on food and glass surfaces. Through proprietary means, O(3) was introduced to produce dry ice pellets to a concentration of 20 parts per million (ppm) by total weight. The ABI sublimation rate was similar to that of dry ice pellets under identical conditions, and ABI was able to hold the O(3) concentration throughout the normal shelf life of the product. Challenge studies were performed using different microorganisms, including E. coli, Campylobacter jejuni, Salmonella, and Listeria, that are critical to food safety. ABI showed significant (P < 0.05) microbial reduction during bioaerosol contamination (up to 5-log reduction of E. coli and Listeria), on chicken breast (approximately 1.3-log reduction of C. jejuni), on contact surfaces (approximately 3.9 log reduction of C. jejuni), and in liquid (2-log reduction of C. jejuni). Considering the stability of O(3), ease of use, and antimicrobial efficacy against foodborne pathogens, our results suggest that ABI is a better alternative, especially for meat and poultry processors, as compared to dry ice. Further, ABI can potentially serve as an additional processing hurdle to guard against pathogens during processing, transportation, distribution, and/or storage.

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 2h, 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.

Use of chemical sanitizers to reduce microbial populations and maintain quality of whole and fresh-cut cantaloupe.

Whole cantaloupes either not inoculated or inoculated with Salmonella Poona were submerged in water, 180 ppm of chlorine, acidified calcium sulfate (ACS: 1.2% Safe(2)O-ACS50), 1,000 ppm of acidified sodium chlorite (ASC), 80 ppm of peroxyacetic acid (PAA), and a combination of ACS and PAA for 10 min. Although only ASC and the combination of ACS and PAA significantly reduced the aerobic plate count of samples taken from the surface of whole cantaloupe (compared with samples taken from cantaloupe submerged in water only), all treatments reduced yeast and mold counts on the whole cantaloupe. However, none of the treatments of whole cantaloupes consistently reduced yeast and mold counts for the samples of fresh-cut cantaloupes. The aerobic plate counts for fresh-cut cantaloupe were reduced by 1 to 2 log CFU/g by sanitization of whole fruit with ASC, ACS, and the combination of ACS and PAA. The low bacterial population on the fresh-cut fruit was maintained during 14 days of storage at 4 degrees C. All treatments had a limited effect on the population of Salmonella, achieving no more than a 1.5-log reduction of the pathogen inoculated on the surface of the whole cantaloupes. Salmonella was nondetectable via direct plating (with a detection limit of 0.4 log CFU/g) in fresh-cut cantaloupes prepared from whole cantaloupes treated with any of the sanitizers. However, after enrichment, Salmonella often was detectable. Color, texture, soluble solids, pH, ascorbic acid, and drip loss of cut cantaloupes were not consistently affected by any of the whole-fruit treatments. Overall, treatments of whole cantaloupe with ASC, ACS, and the combination of ACS and PAA at the concentrations tested permitted a significant reduction in Salmonella and native microflora of whole and cut fruit; however, Salmonella still could be found in cut cantaloupes from all treatments.

Efficacy of chlorine, acidic electrolyzed water and aqueous chlorine dioxide solutions to decontaminate Escherichia coli O157:H7 from lettuce leaves.

This study compared the efficacy of chlorine (20-200 ppm), acidic electrolyzed water (50 ppm chlorine, pH 2.6), acidified sodium chlorite (20-200 ppm chlorite ion concentration, Sanova), and aqueous chlorine dioxide (20-200 ppm chlorite ion concentration, TriNova) washes in reducing populations of Escherichia coli O157:H7 on artificially inoculated lettuce. Fresh-cut leaves of Romaine or Iceberg lettuce were inoculated by immersion in water containing E. coli O157:H7 (8 log CFU/ml) for 5 min and dried in a salad spinner. Leaves (25 g) were then washed for 2 min, immediately or following 24 h of storage at 4 degrees C. The washing treatments containing chlorite ion concentrations of 100 and 200 ppm were the most effective against E. coli O157:H7 populations on Iceberg lettuce, with log reductions as high as 1.25 log CFU/g and 1.05 log CFU/g for TriNova and Sanova wash treatments, respectively. All other wash treatments resulted in population reductions of less than 1 log CFU/g. Chlorine (200 ppm), TriNova, Sanova, and acidic electrolyzed water were all equally effective against E. coli O157:H7 on Romaine, with log reductions of approximately 1 log CFU/g. The 20 ppm chlorine wash was as effective as the deionized water wash in reducing populations of E. coli O157:H7 on Romaine and Iceberg lettuce. Scanning electron microscopy indicated that E. coli O157:H7 that was incorporated into biofilms or located in damage lettuce tissue remained on the lettuce leaf, while individual cells on undamaged leaf surfaces were more likely to be washed away.

Scientific status summary.

The Institute of Food Technologists has issued this Scientific Status Summary to provide readers with a tutorial on biofilms, their purposeful mechanism of interaction (quorum sensing), and recent findings on how to inhibit their formation.

Combination of hot-water surface pasteurization of whole fruit and low-dose gamma irradiation of fresh-cut cantaloupe.

Improvements in methods for disinfecting fresh-cut cantaloupe could reduce spoilage losses and reduce the risk of foodborne illness from human pathogen contamination. The objective of this study was to investigate the feasibility of using hot-water treatment in combination with low-dose irradiation to reduce native microbial populations while maintaining the quality of fresh-cut cantaloupe. Whole cantaloupes were washed in tap water at 20 or 76 degrees C for 3 min. Fresh-cut cantaloupe cubes, prepared from the washed fruit, were then packaged in clamshell containers, and half the samples were exposed to 0.5 kGy of gamma radiation. Native microflora populations and sensory qualities were evaluated during the subsequent 7 days of storage at 4 degrees C. The hot-water surface pasteurization reduced the microflora population by 3.3 log on the surface of whole fruits, resulting in a lower microbial load on the fresh-cut cubes compared with cubes cut from fruit treated with cold water. Irradiation of cubes prepared from untreated fruit to an absorbed dose of 0.5 kGy achieved a low microbial load similar to that of cubes prepared from hot-water-treated fruit. The combination of the two treatments was able to further reduce the microflora population. During storage, the headspace atmosphere of the packages was not significantly influenced by any of the treatments. Color, titratable acidity, pH, ascorbic acid, firmness, and drip loss were not consistently affected by treatment with irradiation, hot water, or the combination of the two. Cubes prepared from hot-water-treated whole fruit had slightly lower soluble solids content. The combination of hot-water pasteurization of whole cantaloupe and low-dose irradiation of packaged fresh-cut melon can reduce the population of native microflora while maintaining the quality of this product.

Biofilm reactors for industrial bioconversion processes: employing potential of enhanced reaction rates.

This article describes the use of biofilm reactors for the production of various chemicals by fermentation and wastewater treatment. Biofilm formation is a natural process where microbial cells attach to the support (adsorbent) or form flocs/aggregates (also called granules) without use of chemicals and form thick layers of cells known as "biofilms." As a result of biofilm formation, cell densities in the reactor increase and cell concentrations as high as 74 gL(-1) can be achieved. The reactor configurations can be as simple as a batch reactor, continuous stirred tank reactor (CSTR), packed bed reactor (PBR), fluidized bed reactor (FBR), airlift reactor (ALR), upflow anaerobic sludge blanket (UASB) reactor, or any other suitable configuration. In UASB granular biofilm particles are used. This article demonstrates that reactor productivities in these reactors have been superior to any other reactor types. This article describes production of ethanol, butanol, lactic acid, acetic acid/vinegar, succinic acid, and fumaric acid in addition to wastewater treatment in the biofilm reactors. As the title suggests, biofilm reactors have high potential to be employed in biotechnology/bioconversion industry for viable economic reasons. In this article, various reactor types have been compared for the above bioconversion processes.

Biofilm formation, cellulose production, and curli biosynthesis by Salmonella originating from produce, animal, and clinical sources.

The ability of 71 strains of Salmonella enterica originating from produce, meat, or clinical sources to form biofilms was investigated. A crystal violet binding assay demonstrated no significant differences in biofilm formation by isolates from any source when tested in any of the following three media: Luria-Bertani broth supplemented with 2% glucose, tryptic soy broth (TSB), or 1/20th-strength TSB. Incubation was overnight at 30 degrees C under static conditions. Curli production and cellulose production were monitored by assessing morphotypes on Luria-Bertani agar without salt containing Congo red and by assessing fluorescence on Luria-Bertani agar containing calcofluor, respectively. One hundred percent of the clinical isolates exhibited curli biosynthesis, and 73% demonstrated cellulose production. All meat-related isolates formed curli, and 84% produced cellulose. A total of 80% of produce-related isolates produced curli, but only 52% produced cellulose. Crystal violet binding was not statistically different between isolates representing the three morphotypes when grown in TSB; however, significant differences were observed when strains were cultured in the two other media tested. These data demonstrate that the ability to form biofilms is not dependent on the source of the test isolate and suggest a relationship between crystal violet binding and morphotype, with curli- and cellulose-deficient isolates being least effective in biofilm formation.

Studies to select appropriate nonpathogenic surrogate Escherichia coli strains for potential use in place of Escherichia coli O157:H7 and salmonella in pilot plant studiest.

The response of a potential nonpathogenic surrogate organism to a particular treatment should closely mimic the response of the target pathogenic organism. In this study, growth characteristics (generation time, lag phase duration, and maximum population), pH at stationary phase, and survival characteristics (level of attachment and survival on apple surfaces, resistance to hydrogen peroxide decontamination treatments, and thermal resistance at 60 degrees C) of 15 nonpathogenic generic Escherichia coli strains and one nonpathogenic E. coli O157:H43 strain were compared with those of two E. coli O157:H7 strains and two Salmonella strains. Few differences in growth characteristics or pH at stationary phase were evident between nonpathogenic and pathogenic strains tested. However, considerably more separation among strains was seen following investigation of survival characteristics. E. coli ECRC 97.0152, which does not contain genes encoding for known virulence factors associated with E. coli O157:H7, appears to be a good surrogate candidate, with growth and survival characteristics similar to those of E. coli O157:H7 strains. The less heat-resistant surrogate strains E. coli NRRL B-766 and NRRL B-3054 and E. coli ATCC 11775, ATCC 25253, and ATCC 25922 may be used when attempting to model the heat resistance of Salmonella Montevideo G4639 and Salmonella Poona RM 2350, respectively. These surrogate strains may be useful for evaluating the efficacy of intervention steps in reducing populations of selected strains of E. coli O157:H7 and Salmonella in processing environments where these pathogens cannot be introduced.