Use of chlorine dioxide gas for the postharvest control of Alternaria alternata and Stemphylium vesicarium on Roma tomatoes.

BACKGROUND: Tomatoes and potatoes are the top produce affected in terms of value lost in the USA. Postharvest losses can occur anywhere from the time of harvest to the consumers' decision to eat or discard the food. These data support the importance of finding sustainable strategies to minimise food waste and preserve resources. This study evaluated the potential application of chlorine dioxide gas (ClO2 ) technology to control the postharvest spoilage of Roma tomatoes by Alternaria alternata and Stemphylium vesicarium. RESULTS: Data analysis showed that exposure time was a significant factor for fungal disease control (P < 0.05). After 3 min of treatment, mycelial growth was completely inhibited for A. alternata and S. vesicarium. Similar results were observed for conidial germination. The efficacy of ClO2 treatments was also studied under in vivo conditions. While untreated Roma tomatoes developed white moulds and black spots after 5 days of storage, produce decay was significantly (P < 0.05) delayed after 5 and 7 min treatments for S. vesicarium and A. alternata respectively. CONCLUSION: The use of ClO2 in the food industry is regulated by both the FDA and the EPA. Currently, only acidified sodium chlorite solutions are approved for the control of micro-organisms in water used to wash fruits and vegetables. No direct applications of ClO2 gas on fresh fruits and vegetables can be found in the regulations. More data are required by the two agencies to demonstrate that residues of ClO2 on produce surfaces are acceptable for human consumption.

Comparing the mannitol-egg yolk-polymyxin agar plating method with the three-tube most-probable-number method for enumeration of Bacillus cereus spores in raw and high-temperature, short-time pasteurized milk.

The U.S. Food and Drug Administration's Bacteriological Analytical Manual recommends two enumeration methods for Bacillus cereus: (i) standard plate count method with mannitol-egg yolk-polymyxin (MYP) agar and (ii) a most-probable-number (MPN) method with tryptic soy broth (TSB) supplemented with 0.1% polymyxin sulfate. This study compared the effectiveness of MYP and MPN methods for detecting and enumerating B. cereus in raw and high-temperature, short-time pasteurized skim (0.5%), 2%, and whole (3.5%) bovine milk stored at 4°C for 96 h. Each milk sample was inoculated with B. cereus EZ-Spores and sampled at 0, 48, and 96 h after inoculation. There were no differences (P > 0.05) in B. cereus populations among sampling times for all milk types, so data were pooled to obtain overall mean values for each treatment. The overall B. cereus population mean of pooled sampling times for the MPN method (2.59 log CFU/ml) was greater (P < 0.05) than that for the MYP plate count method (1.89 log CFU/ml). B. cereus populations in the inoculated milk samples ranged from 2.36 to 3.46 and 2.66 to 3.58 log CFU/ml for inoculated milk treatments for the MYP plate count and MPN methods, respectively, which is below the level necessary for toxin production. The MPN method recovered more B. cereus, which makes it useful for validation research. However, the MYP plate count method for enumeration of B. cereus also had advantages, including its ease of use and faster time to results (2 versus 5 days for MPN).

Effect of chlorine dioxide gas on Salmonella enterica inoculated on navel orange surfaces and its impact on the quality attributes of treated oranges.

Microorganisms, including pathogens of public health significance, have been shown to contaminate orange juice during the mechanical extraction of juice. The problem gets exacerbated when washed oranges have high initial microbial load, due to an insufficient postharvest treatment. The objective of this study was to investigate the reduction of Salmonella enterica on orange surfaces using ClO2 gas treatments to achieve a 5 log reduction, consistent with the recommendations of the U.S. Department of Agriculture-National Advisory Committee on Microbiological Criteria for Foods. A mixed culture of four Salmonella strains, isolated from previous orange juice outbreaks, was spot inoculated onto orange skin surface areas. The oranges were then treated with 0.1, 0.3, and 0.5 mg/L ClO2 gas for 2-14 minutes at 22°C and 90%-95% relative humidity. Surviving bacteria on treated areas were recovered and enumerated over treatment time on a nonselective medium, tryptic soy agar, followed by culturing onto a selective medium, xylose lysine deoxycholate agar. A >5 log reduction of Salmonella per sample of orange surface was observed with 0.1 and 0.3 mg/L ClO2 gas treatments at 14 minutes and a similar log reduction was observed at 0.5 mg/L ClO2 gas at 10 minutes. This result demonstrates that the treatment of oranges with ClO2 gas is a promising technology that could be successfully employed for the treatment of whole oranges to reduce the risk of Salmonella outbreaks in orange juice.

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 log(10) population of biofilm cells before CD gas, aqueous CD and SHC treatment was 4.80, 5.09 and 4.95 log(10) CFU/cm(2). 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 log(10) CFU/cm(2), 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 Salmonella enterica and Listeria monocytogenes inoculated on hydroponic tomatoes using chlorine dioxide gas.

The main objective of this study was to determine survivability of a cocktail of three strains of Salmonella enterica (Montevideo, Javiana, and Baildon) and two strains of Listeria monocytogenes (LCDC 81-861 and F4244) on hydroponic tomatoes after treatment with chlorine dioxide (ClO(2)) gas. An initial concentration of 8-9 log cfu/mL of Salmonella and Listeria cocktails was inoculated individually, in separate experiments, on tomato skin to obtain a population of 7-8 log cfu/cm(2) after drying of the inoculums on the tomato skin. The aim was to achieve a 5 log reduction consistent with the recommendations of the National Advisory Committee on Microbiological Criteria for Foods. The tomato skins were treated with 0.1, 0.3, and 0.5 mg/L ClO(2) gas for 12 min at 22 degrees C and at the relative humidity of 90%. Untreated skin samples were processed under the same conditions. ClO(2)-gas-treated and untreated samples were recovered by an overlay method. The bottom layer contains tryptic soy agar, and the top layer consists of xylose-lysine-desoxycholate agar or modified Oxford antimicrobial supplement agar for Salmonella and Listeria, respectively. More than a 5 log reduction in Salmonella and Listeria was observed on the tomato skin surfaces after treatment with 0.5 mg/L ClO(2) gas for 12 min. Treatment with 0.5 mg/L ClO(2) gas for 12 min also delayed the growth of natural microflora on tomato surfaces and extended the shelf life of tomatoes by 7 days during storage at 22 degrees C, compared with the untreated control. These results revealed that ClO(2) gas is a promising antimicrobial technology for fresh tomato skin surfaces.

Inactivation of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on spinach leaves by X-ray.

Several recent foodborne disease outbreaks associated with leafy green vegetables, including spinach, have been reported. X-ray is a non-thermal technology that has shown promise for reducing pathogenic and spoilage bacteria on spinach leaves. Inactivation of inoculated Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on spinach leaves using X-ray at different doses (0.1, 0.2, 0.3, 0.5, 0.75, 1.0, 1.5 and 2.0 kGy) was studied. The effect of X-ray on color quality and microflora counts (mesophilic counts, psychrotrophic counts and yeast and mold counts) of untreated and treated spinach was also determined. A mixture of three strains of each tested organism was spot inoculated (100 microl) onto the surface of spinach leaves (approximately 8-9 log ml(-1)), separately, and air-dried, followed by treatment with X-ray at 22 degrees C and 55-60% relative humidity. Surviving bacterial populations on spinach leaves were evaluated using a nonselective medium (tryptic soy agar) with a selective medium overlay for each bacteria; E. coli O157:H7 (CT-SMAC agar), L. monocytogenes (MOA), and S. enterica and S. flexneri (XLD). More than a 5 log CFU reduction/leaf was achieved with 2.0 kGy X-ray for all tested pathogens. Furthermore, treatment with X-ray significantly reduced the initial inherent microflora on spinach leaves and inherent levels were significantly (p < 0.05) lower than the control sample throughout refrigerated storage for 30 days. Treatment with X-ray did not significantly affect the color of spinach leaves, even when the maximum dose (2.0 kGy) was used.

Identification of a non-pathogenic surrogate organism for chlorine dioxide (ClO2) gas treatment.

The identification of non-pathogenic surrogate microorganisms is beneficial for determining and validating the efficacy of antimicrobial treatments in food manufacturing environments. A surrogate organism was identified to aid in the decontamination process of fresh produce when treated with chlorine dioxide (ClO(2)) gas. Thirty-two known strains of pathogenic and non-pathogenic microorganisms and seven unknown microbial isolates from mushroom, tomatoes, and strawberries were evaluated. The primary goal was to find alternative non-pathogenic organisms that had an equal or higher resistance compared to Escherichia coli O157:H7, Salmonella spp., and Listeria monocytogenes. Among the strains tested, MR1 (mushroom isolate), E. coli O157:H7 C7927, E. coli O157:H7 204P, STB2 (strawberry isolate), and vegetative cells of Bacillus cereus 232 in wet inoculum were found to be the most resistant to gaseous ClO(2) treatment at 0.3 mg/l for 1 min and D-values at 0.3 mg/l ClO(2) were 3.53, 1.95, 1.72, 1.68, and 1.57 min, respectively. For identification, the MR1 and STB2 strains were identified using a Ribotyper with the EcoRI restriction enzyme of 16S rDNA sequence. MR1 was identified as Hafnia alvei with a similarity value of 94% using the ribotype pattern and with a 93.6% similarity using an API 20E strip, and with a 99% similarity using 16S rDNA analysis. The Ped-2E9-based cytotoxicity assay was conducted for the MRI strain extracellular toxin and whole cell toxicity and did not show cytotoxicity. Analysis, using multiplex PCR, was performed to verify absence of the eaeA gene. H. alvei is a suitable non-pathogenic surrogate, with higher resistance to ClO(2) gas compared to pathogens studied, that may be useful to establish optimum conditions of ClO(2) gas decontamination systems.

Survival and growth of foodborne microorganisms in processed and individually wrapped cheese slices.

The objectives of the research reported here were to determine the growth, survival, or inactivation of selected microorganisms on individually wrapped processed cheese (IWC) slices stored at 5 degrees C and 22 degrees C, and to compare quality indices. IWC slices were spot-inoculated with foodborne pathogenic bacteria (Listeria monocytogenes, Staphylococcus aureus, and Salmonella spp.), spoilage bacteria (Pseudomonas spp. and Lactobacillus spp.), and spoilage molds (Penicillium spp. and Cladosporium spp.). Each bacterium was inoculated at 10(5) CFUs/g for determination of growth, survival, or inactivation. Molds were inoculated at 10(2) spores per gram and observed for growth. Fifty percent of the inoculated product samples were held at 5 degrees C (to simulate refrigeration), and the other 50 percent were held at 22 degrees C (to simulate ambient temperature) throughout shelf life. Samples taken on days 0, 3, 7,10, 14, and 28 and after 2, 3, 6, and 9 months, and were evaluated for surviving cells (by means of appropriate selective media), color (with the cheese color guide), and lipid oxidation (by means of peroxide values). Bacterial inactivation was observed in all conditions. At 14 days, a 5-log reduction was observed for Listeria monocytogenes and Salmonella, while a 3-log reduction was observed for Staphylococcus aureus. For Pseudomonas spp. and Lactobacillus spp., a 2-log reduction was observed within 3 days, with an additional 1-log reduction noted after several months. Mold levels showed no change during the first several weeks of storage. At 84 days, mold levels decreased at 5 degrees C, but they showed growth at 22 degrees C, to approximately 10(5) CFUs/g. Visual color was evaluated on a 10-point National Cheese Institute scale. During storage at 5 degrees C or 22 degrees C, color became darker and values increased from 4 to 5 and 4 to 7, respectively. Higher peroxide values were also obtained for the samples held at 22 degrees C versus 5 degrees C. From a microbiological standpoint, pathogenic and spoilage bacteria were unable to grow in this product; however, long-term storage at 22 degrees C led to lower product quality and mold growth.

Growth and survival of selected pathogens in margarine-style table spreads.

Although margarine-style table spreads can have a pH above 4.6 and a water activity greater than 0.85, there is some question if such products can support the growth of pathogenic bacteria. The objective of this study was to evaluate the growth and survival of Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella typhi in 60-percent- and 70-percent-vegetable-oil, margarine-style, water-in-oil-emulsion table spreads stored at different temperatures. Samples of 25 grams of each table spread were inoculated with 1 x 10(3) cells of each bacterial mixture. The samples were stored at 5 degrees C, 7 degrees C, and 21 degrees C, and the microbial population in colony-forming units per gram (CFU/gram) was enumerated over time. In almost all storage conditions, bacterial levels were shown to decrease over time. Inactivation was observed in (listed from fastest to slowest, respectively) S. aureus, L. monocytogenes, E. coli O157:H7, and S. typhi. Growth was observed only for S. typhi in table spreads stored at 21 degrees C, but the rate of growth was extremely slow. Based on these findings, the table spreads evaluated in this study are not potentially hazardous foods, and cold temperature storage is not necessary from a food safety perspective.