Use of titanium dioxide (TiO2) photocatalysts as alternative means for Listeria monocytogenes biofilm disinfection in food processing

The aim of this work was to study the photocatalytic activity of titanium dioxide (TiO₂) against Listeria monocytogenes bacterial biofilm. Different TiO₂ nanostructured thin films were deposited on surfaces such as stainless steel and glass using the doctor-blade technique. All the surfaces were placed in test tubes containing Brain Heart (BH) broth and inoculated with L. monocytogenes. Test tubes were then incubated for 10 days at 16 °C in order to allow biofilm development. After biofilm formation, the surfaces were illuminated by ultraviolet A light (UVA; wavelength of 315-400 nm). The quantification of biofilms was performed using the bead vortexing method, followed by agar plating and/or by conductance measurements (via the metabolic activity of biofilm cells). The presence of the TiO₂ nanoparticles resulted in a fastest log-reduction of bacterial biofilm compared to the control test. The biofilm of L. monocytogenes for the glass nanoparticle 1 (glass surface modified by 16% w/v TiO₂) was found to have decreased by 3 log CFU/cm² after 90 min irradiation by UVA. The use of TiO₂ nanostructured photocatalysts as alternative means of disinfecting contaminated surfaces presents an intriguing case, which by further development may provide potent disinfecting solutions. Surface modification using nanostructured titania and UV irradiation is an innovative combination to enhance food safety and economizing time and money.     

Comparison of inactivation of Listeria monocytogenes within a biofilm matrix using chlorine dioxide gas,aqueous chlorine dioxide and sodium hypochlorite treatments

The present research compared the effect of chlorine dioxide (CD) gas, aqueous CD and aqueous sodium hypochlorite (SHC) treatments on the inactivation of a five strain mixture of Listeria monocytogenes – containing biofilms. Four day old biofilms were developed on a stainless steel (SS 304) coupon by using a mixture of five cultures of L. monocytogenes (Scott A, N1-227, 103M, 82 and 311) using a 100% relative humidity (RH) dessicator for incubation at room temperature (22 ± 2 °C). After biofilm development, coupons were rinsed and dried for 2 h and treated with 0.3 mg/l CD gas at 75% RH, 7 mg/l of aqueous CD and 50 mg/l SHC. Initial log₁₀ population of biofilm cells before CD gas, aqueous CD and SHC treatment was 4.80, 5.09 and 4.95 log₁₀ CFU/cm². The Weibull model was used to fit non-linear survivor curves. Treatments and time points of 0.3 mg/l CD gas and 7 mg/l aq. CD solution were significantly different (p < 0.05). A 10 min treatment of 0.3 mg/l CD gas, 7 mg/l of aq. CD, and 50 mg/l SHC resulted in reductions of 3.21, 3.74 and 3.09 log₁₀ CFU/cm², respectively. At 10 min, all treatments were not statistically different (p > 0.05). Low levels of CD (0.3 mg/l CD gas and 7 mg/l aq. CD solution) for 10 min resulted in similar log reductions compared to 50 mg/l SHC.     

Inactivation of biofilm cells of foodborne pathogens by steam pasteurization

The objective of this study was to evaluate the effect of steam pasteurization on the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms on stainless steel and polyvinyl chloride (PVC). Biofilms were formed on a stainless steel and PVC coupon by using a mixture of three strains each of three foodborne pathogens. Six-day-old biofilms on stainless steel and PVC coupons were treated with steam at 75 and 85 °C for 5, 10, 20, 30, 40, and 50 s. Biofilm cells of E. coli O157:H7, S. Typhimurium, and L. monocytogenes on stainless steel were reduced by more than 6 log CFU/coupon after exposure to steam at 75 °C for 30, 40, and 30 s, respectively, and at 85 °C for 30, 20, and 20 s, respectively. Steam treatment resulted in less reduction in the levels of biofilm cells on PVC coupons. Biofilm cells of E. coli O157:H7, S. Typhimurium, and L. monocytogenes were reduced by 1.78, 2.04, and 1.29 log CFU/coupon, respectively, after 50 s of exposure to steam at 75 °C. Exposure to steam at 85° for 50 s reduced biofilm cells of E. coli O157:H7, S. Typhimurium, and L. monocytogenes by 2.53, 3.01, and 1.70 log CFU/coupon, respectively. The results of this study suggest that steam pasteurization has potential as a biofilm control method by the food industry.     

Resistance of pathogenic bacteria on the surface of stainless steel depending on attachment form and efficacy of chemical sanitizers

Various bacteria including food spoilage bacteria and pathogens can form biofilms on different food processing surfaces, leading to potential food contamination or spoilage. Therefore, the survival of foodborne pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, Cronobacter sakazakii) in different forms (adhered cells, biofilm producing in TSB, biofilm producing at RH 100%) on the surface of stainless steel and stored at various relative humidities (RH 23%, 43%, 68%, 85%, and 100%) at room temperature for 5 days was investigated in this study. Additionally, the efficacy of chemical sanitizers (chlorine-based and alcohol-based commercial sanitizers) on inhibiting various types of biofilms of E. coli O157:H7 and S. aureus on the surface of stainless steel was investigated. The number of pathogens on the surface of stainless steel in TSB stored at 25 °C for 7 days or RH 100% at 25 °C for 7 days was significantly increased and resulted in the increase of 3 log₁₀ CFU/coupon after 1 day, and these levels were maintained for 7 days. When stainless steel coupons were stored at 25 °C for 5 days, the number of pathogens on the surface of stainless steel was significantly reduced after storage at RH 23%, 43%, 68%, and 85%, but not at 100%. When the bacteria formed biofilms on the surface of stainless steel in TSB after 6 days, the results were similar to those of the attached form. However, levels of S. aureus and C. sakazakii biofilms were more slowly reduced after storage at RH 23%, 43%, 68%, and 85% for 5 days than were those of the other pathogens. Formation of biofilms stored at RH 100% for 5 days displayed the highest levels of resistance to inactivation. Treatment with the alcohol sanitizer was very effective at inactivating attached pathogens or biofilms on the surface of stainless steel. Reduction levels of alcohol sanitizer treatment ranged from 1.91 to 4.77 log and from 4.35 to 5.35 log CFU/coupon in E. coli O157:H7 and S. aureus, respectively. From these results, the survival of pathogens contaminating the surfaces of food processing substrates such as stainless steel varied depending on RH and attachment form. Also, alcohol-based sanitizers can be used as a potential method to remove microbial contamination on the surfaces of utensils, cooking equipment, and other related substrates regardless of the microbial attached form.     

Inactivation of Escherichia coli O157:H7 on stainless steel upon exposure to Paenibacillus polymyxa biofilms

We investigated the potential use of biofilm formed by a competitive-exclusion (CE) microorganism to inactivate Escherichia coli O157:H7 on a stainless steel surface. Five microorganisms showing inhibitory activities against E. coli O157:H7 were isolated from vegetable seeds and sprouts. The microorganism with the greatest antimicrobial activity was identified as Paenibacillus polymyxa (strain T5). In tryptic soy broth (TSB), strain T5 reached a higher population at 25 °C than at 12 or 37 °C without losing inhibitory activity against E. coli O157:H7. When P. polymyxa (6 log CFU/mL) was co-cultured with E. coli O157:H7 (2, 3, 4, or 5 log CFU/mL) in TSB at 25 °C, the number of E. coli O157:H7 decreased significantly within 24 h. P. polymyxa formed a biofilm on stainless steel coupons (SSCs) in TSB at 25 °C within 24 h, and cells in biofilms, compared to attached cells without biofilm formation, showed significantly increased resistance to a dry environment (43% relative humidity [RH]). With the exception of an inoculum of 4 log CFU/coupon at 100% RH, upon exposure to biofilm formed by P. polymyxa on SSCs, populations of E. coli O157:H7 (2, 4, or 6 log CFU/coupon) were significantly reduced within 48 h. Most notably, when E. coli O157:H7 at 2 log CFU/coupon was applied to SSCs on which P. polymyxa biofilm had formed, it was inactivated within 1 h, regardless of RH. These results will be useful when developing strategies using biofilms produced by competitive exclusion microorganisms to inactivate foodborne pathogens in food processing environments.     

Attachment and biofilm formation by Escherichia coli O157:H7 at different temperatures, on various food-contact surfaces encountered in beef processing

Escherichia coli O157:H7 attached to beef-contact surfaces found in beef fabrication facilities may serve as a source of cross-contamination. This study evaluated E. coli O157:H7 attachment, survival and growth on food-contact surfaces under simulated beef processing conditions. Stainless steel and high-density polyethylene surfaces (2 × 5 cm) were individually suspended into each of three substrates inoculated (6 log CFU/ml or g) with E. coli O157:H7 (rifampicin-resistant, six-strain composite) and then incubated (168 h) statically at 4 or 15 °C. The three tested soiling substrates included sterile tryptic soy broth (TSB), unsterilized beef fat-lean tissue (1:1 [wt/wt]) homogenate (10% [wt/wt] with sterile distilled water) and unsterilized ground beef. Initial adherence/attachment of E. coli O157:H7 (0.9 to 2.9 log CFU/cm²) on stainless steel and high-density polyethylene was not affected by the type of food-contact surface but was greater (p < 0.05) through ground beef. Adherent and suspended E. coli O157:H7 counts increased during storage at 15 °C (168 h) by 2.2 to 5.4 log CFU/cm² and 1.0 to 2.8 log CFU/ml or g, respectively. At 4 °C (168 h), although pathogen levels decreased slightly in the substrates, numbers of adherent cells remained constant on coupons in ground beef (2.4 to 2.5 log CFU/cm²) and increased on coupons in TSB and fat-lean tissue homogenate by 0.9 to 1.0 and 1.7 to 2.0 log CFU/cm², respectively, suggesting further cell attachment. The results of this study indicate that E. coli O157:H7 attachment to beef-contact surfaces was influenced by the type of soiling substrate and temperature. Notably, attachment occurred not only at a temperature representative of beef fabrication areas during non-production hours (15 °C), but also during cold storage (4 °C) temperatures, thus, rendering the design of more effective sanitation programs necessary.     

Use of high-concentration-short-time chlorine dioxide gas treatments for the inactivation of Salmonella enterica spp. inoculated onto Roma tomatoes

Salmonella outbreaks have been recently linked to the consumption of fresh tomatoes. Thus, there is a need to develop systems that reduce the risk of microbial contamination to increase product shelf-life and keep fresh fruit attributes. The objectives of this study were to evaluate high-concentration-short-time chlorine dioxide gas treatments effects on Salmonella-inoculated Roma tomatoes and determine the optimal treatment conditions for microbial inactivation and shelf-life extension. Effects of ClO₂ concentration (2, 5, 8 and 10 mg/l) and exposure time (10, 30, 60, 120 and 180 s) on inoculated Roma tomatoes were studied. Salmonella enterica strains, serotype Montevideo, Javiana and Baildon, were used to experimentally inoculate the food product. After ClO₂ treatments, tomatoes were stored at room temperature for 28 days. Inherent microbial population, change in tomato color, and chlorine dioxide gas residuals were evaluated. ANOVA analysis showed that both ClO₂ concentration and treatment time were significant (p < 0.01) for Salmonella inactivation. Surviving Salmonella populations of 3.09, 2.17 and 1.16 log CFU/cm² were obtained treating tomatoes with 8 mg/l ClO₂ for 60 s, 10 mg/l ClO₂ for 120 s, and 10 mg/l for 180 s, respectively (initial Salmonella population: 6.03 ± 0.11 log CFU/cm²). The selected treatments significantly reduced background microflora (p < 0.05), while fruit color and residual contents were not significantly different (p > 0.05), as compared to the control. Results suggest the potential for high-concentration-short-time treatments ClO₂ gas as an effective pathogen inactivation technology for large-scale produce packing operations.     

Effect of reduced oxygen atmosphere and sodium acetate treatment on the microbial quality changes of seer fish (Scomberomorus commerson) steaks stored in ice

The effect of reduced oxygen atmosphere and sodium acetate treatment on the microbial quality of seer fish (Scomberomorus commerson) steaks was determined during chilled storage (1–2 °C). The O₂ absorber reduced the oxygen content in the pack to less than 0.01% corresponding to 99.96% reduction within 24 h. The use of O₂ absorber with sodium acetate dip treatment (2% w/v) extended the sensory shelf life up to 25 days compared to only 12 days for control air packs and 20 days for untreated samples with O₂ absorber. A prominent lag phase was observed for many bacterium studied, particularly for the sodium acetate treated samples with O₂ absorber. On the day of sensory rejection, both the total mesophilic and psychrotrophic counts reached 7.7–8.1 and 7.1–7.9 log cfu/g, respectively. The sodium acetate treatment and reduced O₂ atmosphere affected the type of major spoilers. In air packed samples, H₂S-producers predominated followed by Brochothrix thermosphacta, Pseudomonas spp., where as in the untreated samples with O₂ absorber, H₂S-producers predominated the microbial flora followed by Lactobacillus spp. For treated samples with O₂ absorber, B. thermosphacta formed the major micro-flora followed by Lactobacillus spp. The use of O₂ absorber inhibited the growth of Pseudomonas spp., and total Enterobacteriaceae.     

Biofilm formation and exopolysaccharide (EPS) production by Cronobacter sakazakii depending on environmental conditions

Biofilm matrices are formed largely of extracellular polymeric substance (EPS). This study was conducted to investigate biofilm formation and EPS production by Cronobacter sakazakii under various conditions (media, nutrition, and relative humidity (RH)) by quantification of EPS and cell populations, Field Emission Scanning Electron Microscope (FE-SEM), and colony observation. Various agar media conditions (TSA without dextrose (W/D), M9 minimum salt medium (MSM) agar, and M9 MSM agar with 3% glucose, 3% NaCl, 3% Tween 80, 3% sucrose, and adjusted to pH 5 with HCl) were prepared. C. sakazakii biofilm formed on the surface of stainless steel coupons (SSCs) immersed in TSB W/D and M9 MSM with or without 0, 1, 3, and 5% sucrose and subsequently exposed to various RH levels (23, 43, 68, 85, and 100%). EPS production by C. sakazakii on TSA W/D was significantly higher than that on other media after 1 and 2 days. However, C. sakazakii ATCC 12868 produced the highest levels of EPS (209.18 ± 16.13 and 207.22 ± 4.14 μg/mL after 1 and 2 days, respectively) on M9 MSM agar with 3% sucrose. Regarding C. sakazakii ATCC 12868 biofilm formed on the surface of SSCs immersed in M9 MSM with 0, 1, 3, and 5% sucrose and subsequently exposed to various RHs, populations were significantly different among the various RHs and sucrose concentrations, and EPS production was significantly higher (4.69 mg/L) compared to other sucrose concentrations (0%:0.71 mg/L and 1%:0.98 mg/L), except for M9 MSM with 3% sucrose (2.97 mg/L) (P ≤ 0.05). From these results, biofilm formation and EPS production by C. sakazakii differed depending on the nutrient or environmental conditions provided to the cells.     

Differences in biofilm formation of produce and poultry Salmonella enterica isolates and their persistence on spinach plants

Spinach plants were irrigated biweekly with water containing 2.1 log CFU Salmonella/100 ml water (the maximum Escherichia coli MPN recommended by the Leafy Greens Marketing Agreement; LGMA), or 4.1 CFU Salmonella/100 ml water to determine Salmonella persistence on spinach leaves. Green Fluorescent protein expressing Salmonella were undetectable by most-probable number (MPN) at 24 h and 7 days following each irrigation event. This study indicates that Salmonella are unlikely to persist on spinach leaves when irrigation water is contaminated at a level below the LGMA standards. In a parallel study, persistence of Salmonella isolated from poultry or produce was compared following biweekly irrigation of spinach plants with water containing 6 log CFU Salmonella/100 ml. Produce Salmonella isolates formed greater biofilms on polystyrene, polycarbonate and stainless steel surfaces and persisted at significantly higher numbers on spinach leaves than those Salmonella from poultry origin during 35 days study. Poultry Salmonella isolates were undetectable (<1 log CFU/g) on spinach plants 7 days following each irrigation event when assayed by direct plating. This study indicates that Salmonella persistence on spinach leaves is affected by the source of contamination and the biofilm forming ability of the strain.     

Inactivation of biofilm cells of foodborne pathogen by aerosolized sanitizers

The objective of this study was to determine the effect of aerosolized sanitizers on the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms. Biofilms were formed on a stainless steel and polyvinyl chloride (PVC) coupon by using a mixture of three strains each of three foodborne pathogens. Six day old biofilms on stainless steel and PVC coupons were treated with aerosolized sodium hypochlorite (SHC; 100 ppm) and peracetic acid (100, 200, and 400 ppm) in a model cabinet for 5, 10, 30, and 50 min. Treatment with 100 ppm PAA was more effective than the same concentration of SHC with increasing treatment time. Exposure to 100 ppm SHC and PAA for 50 min significantly (p<0.05) reduced biofilm cells of three foodborne pathogens (0.50 to 3.63 log CFU/coupon and 2.83 to more than 5.78 log CFU/coupon, respectively) compared to the control treatment. Exposure to 200 and 400 ppm PAA was more effective in reducing biofilm cells. Biofilm cells were reduced to below the detection limit (1.48 log CFU/coupon) between 10 and 30 min of exposure. The results of this study suggest that aerosolized sanitizers have a potential as a biofilm control method in the food industry.     

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.     

Biofilm formation of Salmonella Typhimurium on stainless steel and acrylic surfaces as affected by temperature and pH level

The effect of temperature (28, 37 and 42 °C) and pH (6 and 7) on the biofilm formation capability of Salmonella Typhimurium on stainless steel and acrylic was investigated. The rate of biofilm formation increased with increasing temperature and pH, while the number of attached cells after 240 h decreased with increasing temperature and was not different between pH 6 and 7. The surface hydrophobicity of bacterial cells was not significantly (p > 0.05) different among tested conditions. Electron-donating/accepting properties changed with pH and temperature, although these changes did not correlate with the ability to form biofilms under respective conditions. Attachment of S. Typhimurium showed a preference for stainless steel compared to acrylic surfaces under all conditions tested. The results suggest that salmonellae were less adherent to acrylic than to stainless steel surfaces; thus, acrylic-type surfaces should be considered for use in the food industry over stainless steel where applicable. The rate of biofilm formation increased at higher temperatures and pH levels within the tested ranges. Hurdle technology using lower temperatures reduced pH may help delay biofilm formation on food contact surfaces contaminated with S. Typhimurium.     

Production of biofilm and quorum sensing by Escherichia coli O157:H7 and its transfer from contact surfaces to meat,poultry, ready-to-eat deli,and produce products

Multistate outbreaks of Escherichia coli O157:H7 infections through consumption of contaminated foods including produce products have brought a great safety concern. The objectives of this study were to determine the effect of biofilm and quorum sensing production on the attachment of E. coli O157:H7 on food contact surfaces and to evaluate the transfer of the pathogen from the food contact to various food products. E. coli O157:H7 produced maximum levels of AI-2 signals in 12 h of incubation in tested meat, poultry, and produce broths and subsequently formed strong biofilm in 24 h of incubation. In general, E. coli O157:H7 formed stronger biofilm on stainless steel than glass. Furthermore, E. coli O157:H7 that had attached on the surface of stainless steel was able to transfer to meat, poultry, ready-to-eat deli, and produce products. Strong attachment of the transferred pathogen on produce products (cantaloupe, lettuce, carrot, and spinach) was detected (>10³ CFU/cm²) even after washing these products with water. Our findings suggest that biofilm formation by E. coli O157:H7 on food contact surfaces can be a concern for efficient control of the pathogen particularly in produce products that require no heating or cooking prior to consumption.     

Inactivation behavior of Pseudomonas aeruginosa by supercritical N₂O compared to supercritical CO₂

Interest is growing for a non-thermal sterilization technique in the food and pharmaceutical industries in order to ensure microbiological safety without the deterioration of product quality. In this study, supercritical nitrous oxide (SC N₂O) treatment was carried out in a multi-batch system to examine its bactericidal effect and characteristics, which largely remains unclear. The effect of operating pressure, temperature, mixing intensity, and working volume ratio (defined as the ratio of sample volume to the reactor volume to be filled with SC fluids) on the inactivation efficiency of SC N₂O were investigated in comparison with supercritical carbon dioxide (SC CO₂) treatment. Pseudomonas aeruginosa was chosen as a model microorganism. A 8-log reduction of P. aeruginosa cell concentration (10% working volume) in neutral phosphate-buffered saline was achieved by the SC N₂O treatment accompanying no pH change in the presence of vigorous mixing (600 rpm) within 6 min, in a condition of 37 °C and 10 MPa and its overall efficiency is comparable to the SC CO₂ treatment. Among the reaction parameters investigated in this study, mixing intensity appeared to be the most important operating parameter affecting the bactericidal efficiency.The release of intracellular substances in cells as a result of the SC N₂O treatment was explained as one of major bactericidal actions by the SC N₂O treatment. At the same time, relatively negligible change of proteins or enzyme activities in the cells in case of the SC N₂O treatment was observed compared to the SC CO₂ treatment. Also, the observation by scanning and transmission electron microscopy shows the milder morphological change of SC N₂O-treated cells in comparison with the SC CO₂-treated cells. This study suggests that the SC N₂O bactericidal application on food products would be a viable option over SC CO₂ application, when mild change of enzyme activity and the proteins, or no pH change are desired.     

A comparative study on the effectiveness of chlorine dioxide gas, ozone gas and e-beam irradiation treatments for inactivation of pathogens inoculated onto tomato, cantaloupe and lettuce seeds

The increase in reported food-borne outbreaks linked with consumption of raw fruits and vegetables has motivated new research focusing on prevention of pre-harvest produce contamination. This study evaluates and compares the effectiveness of three non-thermal technologies, chlorine dioxide gas, ozone gas and e-beam irradiation, for inactivation of Salmonella enterica and Escherichia coli O157:H7 on pre-inoculated tomato, lettuce and cantaloupe seeds, and also their corresponding effect on seeds germination percentage after treatments. Samples were treated with 10 mg/l ClO₂ gas for 3 min at 75% relative humidity, with 4.3 mg/l ozone gas for 5 min and with a dose of 7 kGy electron beam for 1 min. Initial load of pathogenic bacteria on seeds was ~ 6 log CFU/g. Results demonstrate that all treatments significantly reduce the initial load of pathogenic bacteria on seeds (p < 0.05). In particular, after ozone gas treatments 4 log CFU/g reduction was always observed, despite the seeds and/or microorganisms treated. ClO₂ and e-beam treatments were noticeably more effective against Salmonella on contaminated tomato seeds, where 5.3 and 4.4 log CFU/g reduction were respectively observed. Germination percentage was not affected, except for cantaloupe seeds, where the ratio was significantly lowered after ClO₂ treatments. Overall, the results obtained show the great applicability of these non-thermal inactivation techniques to control and reduce pathogenic bacteria contamination of seeds.     

The application of high-concentration short-time chlorine dioxide treatment for selected specialty crops including Roma tomatoes (Lycopersicon esculentum), cantaloupes (Cucumis melo ssp. melo var. cantaloupensis) and strawberries (Fragaria × ananassa)

The effects of high-concentration short-time chlorine dioxide (ClO₂) gas treatment on food-borne pathogens inoculated onto the surface of tomatoes, cantaloupes, and strawberries were studied. Produce were spot-inoculated with a mixture of Salmonella enterica (serotypes Montevideo, Javiana and Baildon), Escherichia coli O157:H7 (serotypes 204 P, EDL 933 and C792) or Listeria monocytogenes (serotypes Scott A, F 5069 and LCDC 81-861), and treated with ClO₂ gas at 10 mg/l for 180 s. After ClO₂ gas treatment, surviving populations were determined and shelf-life studies were conducted (microbial spoilage population, change in color and overall appearance). Significant microbial reduction (p < 0.05) was observed for all treated samples. Nearly a 5 Log CFU/cm²Salmonella reduction was found on tomatoes, cantaloupe and strawberries, while a ∼3 Log CFU/cm² reduction was observed for E. coli and Listeria on all produce surfaces. E. coli and Listeria appeared to be more resistant to ClO₂ gas as compared to Salmonella spp. Treatments significantly (p < 0.05) reduced initial microflora population, while produce color surface was not significantly influenced, as compared to the control (p > 0.05). Results obtained suggest the potential use of high-concentration short-time ClO₂ gas treatment as an effective online pathogen inactivation technology for specialty crops in large-scale produce packing operations.     

Influence of light exposure during storage on the content of sesquiterpene lactones and photosynthetic pigments in witloof chicory (Cichorium intybus L. var. foliosum Hegi)

Greening of witloof chicory is a major quality defect being commonly associated with increased bitterness. The aim of this study was to assess the effect of light exposure during storage on levels of bitter sesquiterpene lactones (SLs) and photosynthetic pigments in leaves and central axes of two cultivars of witloof chicory. For this purpose, witloof chicory packaged in consumer-sized polyamide bags was stored in the dark (8 °C) and under light exposure (8 °C, 4500 lux), respectively, over a period of 11 days. Throughout storage, O₂ and CO₂ levels in the modified atmosphere were monitored. Levels of six individual SLs, chlorophyll derivatives and β-carotene in leaves and central axes were assessed by HPLC-DAD-MSⁿ and color measurements were carried out applying the CIE L*a*b* system.     

Removal of Salmonella enterica serovar Typhimurium and Cronobacter sakazakii biofilms from food contact surfaces through enzymatic catalysis

Bacterial biofilms are highly difficult to control, hence significant economic resources have been allocated to develop strategies to eradicate them. This study evaluated the effect of an enzymatic treatment to be used as a cleaning product to control the presence of biofilms. Two different materials used in the food industry, polystyrene and stainless steel, were tested using Salmonella Typhimuirum and Cronobacter sakazakii. Biofilm formation was carried out by inoculating the surfaces with a standardized concentration of 4 log (CFU cm⁻²) and incubated for 48 hr with renewal of nutrients. The biofilm formation and subsequent enzymatic treatment were quantified using fluorescence microscopy and the conventional culture method. The enzymatic treatment showed significant reductions of 2–3 log (CFU cm⁻²) in biofilm cells, which was attributed to the degradation of the extracellular matrix and the further detachment of both microorganisms. The maximum biofilm detachment obtained with the preventive formula was 46.67%; however, this percentage could be increased by applying an aggressive treatment or by adding a subsequent disinfection step that would eliminate adhered microbial cells. Further, the enzymatic cleaning treatment could be exploited as a potent technology to control bacterial adherence and biofilm formation in the food industry.     

Effect of meat ingredients (sodium nitrite and erythorbate) and processing (vacuum storage and packaging atmosphere) on germination and outgrowth of Clostridium perfringens spores in ham during abusive cooling

The effect of nitrite and erythorbate on Clostridium perfringens spore germination and outgrowth in ham during abusive cooling (15 h) was evaluated. Ham was formulated with ground pork, NaNO₂ (0, 50, 100, 150 or 200 ppm) and sodium erythorbate (0 or 547 ppm). Ten grams of meat (stored at 5 °C for 3 or 24 h after preparation) were transferred to a vacuum bag and inoculated with a three-strain C. perfringens spore cocktail to obtain an inoculum of ca. 2.5 log spores/g. The bags were vacuum-sealed, and the meat was heat treated (75 °C, 20 min) and cooled within 15 h from 54.4 to 7.2 °C. Residual nitrite was determined before and after heat treatment using ion chromatography with colorimetric detection. Cooling of ham (control) stored for 3 and 24 h, resulted in C. perfringens population increases of 1.46 and 4.20 log CFU/g, respectively. For samples that contained low NaNO₂ concentrations and were stored for 3 h, C. perfringens populations of 5.22 and 2.83 log CFU/g were observed with or without sodium erythorbate, respectively. Residual nitrite was stable (p > 0.05) for both storage times. Meat processing ingredients (sodium nitrite and sodium erythorbate) and their concentrations, and storage time subsequent to preparation of meat (oxygen content) affect C. perfringens spore germination and outgrowth during abusive cooling of ham.