Influence of diluent on antimicrobial activity of cinnamon bark essential oil vapor against Staphylococcus aureus and Salmonella enterica on a laboratory medium and beef jerky.

The influence of chemical diluents on the antimicrobial activity of plant essential oil (EO) vapors was evaluated. We first determined if vapors generated from 22 chemical diluents not containing EO had antimicrobial activities. Ethyl ether vapor retarded the growth of S. aureus. The minimal inhibitory concentrations (MICs) and the minimal lethal concentrations (MLCs) of cinnamon bark EO vapor, which was diluted in and generated from 21 diluents, against S. aureus and S. enterica were determined. Cinnamon bark EO vapor showed significantly (P ≤ 0.05) lower MICs against S. aureus when diluted in dimethyl sulfoxide (DMSO), ethanol, ethyl acetate, or jojoba oil, and against S. enterica when diluted in DMSO, ethanol, or jojoba oil, compared to those in other diluents. We compared antimicrobial activities of cinnamon bark EO vapor diluted in DMSO, ethanol, ethyl acetate, or jojoba oil against S. aureus and S. enterica on beef jerky as a food model. Antimicrobial activity was significantly (P ≤ 0.05) higher when vaporized from DMSO. These results indicate that antimicrobial activity of cinnamon bark EO vapor may vary significantly (P ≤ 0.05) depending on the type of diluent from which it is vaporized. These observations provide basic information when developing food and food-contact surface decontamination strategies using EO vapors.

Effects of temperature, pH and sodium chloride on antimicrobial activity of magnesium oxide nanoparticles against E. coli O157:H7.

AIM: This study was done to determine the effects of temperature, pH and sodium chloride (NaCl) on antimicrobial activity of magnesium oxide (MgO) nanoparticles (NPs) against E. coli O157:H7. METHODS AND RESULTS: Culture conditions were established by varying the pH (5.0, 7.2 and 9.0), NaCl concentration (0.5, 2.0, 3.5 and 5.0%, w/v), and incubation temperatures (4, 12, 22 and 37°C). At each condition, the antimicrobial activities of MgO-NPs (0, 1, 2 and 4 mg/ml) against E. coli O157:H7 were measured. Four-way analysis of variance indicated interactions among all factors had a significant effect (p ≤ 0.05) on the antimicrobial activity of MgO-NPs. The concentration of MgO-NPs necessary to cause a 5-log reduction of E. coli O157:H7 under the most inhibitory conditions (37°C, pH 9.0, and 5.0% NaCl) was 0.50 mg/ml of MgO-NPs. CONCLUSION: The antimicrobial activity of the MgO-NPs increased significantly (p ≤ 0.05) with increased temperature, pH and NaCl concentration in TSB. SIGNIFICANCE AND IMPACT OF THE STUDY: The influence of intrinsic and extrinsic factors on antimicrobial activity of MgO-NPs we found will contribute to the development of microbial decontamination strategies using MgO in the food industry.

Antimicrobial activities of plant essential oil vapours against Acidovorax citrulli and Xanthomonas campestris on Cucurbitaceae, Brassicaceae and Solanaceae seeds.

AIM: This study was done to develop a seed decontamination treatment for organic seeds against plant pathogens (Acidovorax citrulli and Xanthomonas campestris) using essential oil (EO) vapours without affecting the seeds' germination rate. METHODS AND RESULTS: By using a diffusion assay and determining minimum inhibitory and lethal concentrations, we screened two EO vapours which were most inhibitory to A. citrulli (cinnamon bark and garlic EO vapours) and X. campestris (onion and garlic EO vapours). After 48 h of exposure to EO vapours at 25°C and 43% or 85% relative humidity (RH), no significant decrease (p > 0.05) in germination rates was observed compared with those of control seeds. It was observed that EO vapour treatment at 25°C and 43% or 85% RH for 48 h caused significant population reductions (p ≤ 0.05) (ca. 0.3-2.6 log colony forming unit/g) compared to those of untreated seeds. CONCLUSION: Applications of EO vapours showed significant (p ≤ 0.05) antimicrobial effects against A. citrulli and X. campestris on both laboratory mediums and plant seeds without decreasing the germination rate of seeds. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides useful information for the development of natural seed sterilization treatments using EO vapours.

Inactivation of Escherichia coli O157:H7 on radish and cabbage seeds by combined treatments with gaseous chlorine dioxide and heat at high relative humidity.

This study was done to develop a method to inactivate Escherichia coli O157:H7 on radish and cabbage seeds using simultaneous treatments with gaseous chlorine dioxide (ClO2) and heat at high relative humidity (RH) without decreasing seeds' viability. Gaseous ClO2 was spontaneously vaporized from a solution containing hydrochloric acid (HCl, 1 N) and sodium chlorite (NaClO2, 100,000 ppm). Using a sealed container (1.8 L), an equation (y = 5687×, R2 = 0.9948) based on the amount of gaseous ClO2 generated from HCl-NaClO2 solution at 60 °C and 85% RH was developed. When radish or cabbage seeds were exposed to gaseous ClO2 at concentrations up to 3,000 ppm for 120 min, germination rates did not significantly decrease (P > 0.05). When seeds inoculated with E. coli O157:H7 were treated with 2,000 or 3,000 ppm of gaseous ClO2 in an atmosphere with 85% RH at 60 °C, populations (6.8-6.9 log CFU/g) on both types of seeds were decreased to below the detection limit for enrichment (-0.5 log CFU/g) within 90 min. This study provides useful information for developing a decontamination method to control E. coli O157:H7 and perhaps other foodborne pathogens on plant seeds by simultaneous treatment with gaseous ClO2 and heat at high RH.

Survival of Salmonella and Shiga Toxin-Producing Escherichia coli and Changes in Indigenous Microbiota during Fermentation of Home-Brewed Kombucha.

ABSTRACT: Survival and growth of Salmonella and Shiga toxin-producing Escherichia coli (STEC) were investigated in kombucha prepared from four brands of commercially available kombucha kits intended for use by home brewers. Changes in populations of the indigenous microbiota responsible for fermentation of kombucha were also determined. An initial population of Salmonella (6.77 log CFU/mL) decreased to below the detection limit (0.30 log CFU/mL) within 10 days in kombucha prepared from two of the test brands. Populations of 1.85 and 1.20 log CFU/mL were detected in two brands fermented for 14 days. An initial STEC population of 7.02 log CFU/mL decreased to <0.30 log CFU/mL in two brands within 14 days; 0.77 and 0.87 log CFU/mL were detected in kombucha prepared from the other two brands. Salmonella and STEC increased within 1 day in three brands of base tea used to prepare kombucha and were stable throughout 14 days of incubation. Both pathogens steadily declined in base tea prepared from one brand of kombucha kit. Inactivation of the pathogens occurred as the pH of the kombucha decreased, but a clear correlation between rates of inactivation among different brands of kits and decrease in pH was not evident. Growth and peak populations of mesophilic aerobic microorganisms, yeasts, lactic acid bacteria, and acetic acid bacteria varied depending on the kombucha kit brand. No strong evidence was found of a correlation between the behavior of Salmonella or STEC and that of any of these groups of indigenous microbiota. Results of this study show that survival of Salmonella and STEC in kombucha and base tea used to prepare kombucha is dependent on inherent differences in commercially available kombucha kits intended for use in home settings. Strict application of hygienic practices is essential for preventing contamination with Salmonella or STEC and reducing the risk of illness associated the consumption of kombucha.

Antimicrobial activities of organic acid vapors against Acidovorax citrulli, Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes on Cucurbitaceae seeds.

This study investigated the antimicrobial activities of organic acid vapors against a phytopathogen (Acidovorax citrulli) and foodborne pathogens (Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes) on the surface of Cucurbitaceae seeds. Germination percentages of cucumber, honeydew melon and watermelon seeds treated with acetic and propionic acid vapors (100 mg/L) at 50 °C and 43% or 85% relative humidity (RH) for up to 2 h did not significantly (P > 0.05) decrease. Treatment with formic acid significantly (P ≤ 0.05) decreased the germination percentage. The antimicrobial activities of acetic and propionic acid vapors (100 mg/L; 50 °C; 43% or 85% RH) were determined. A. citrulli was inactivated within 1 h on cucumber and watermelon seeds, regardless of type of organic acid or RH. The phytopathogen was reduced to levels below the detection limit (-0.5 log CFU/g) for enrichment on honeydew melon seeds treated with acetic acid vapor. S. enterica and L. monocytogenes were inactivated within 2 h at 85% RH on honeydew melon and watermelon seeds treated with acetic acid and propionic acid vapors. E. coli O157: H7 was inactivated by treatment with acetic acid vapor at 85% RH. This study provides useful information for developing a method to decontaminate Curcurbitaceae seeds using organic acid vapors as lethal agents.

Synergistic antimicrobial activity of oregano and thyme thymol essential oils against Leuconostoc citreum in a laboratory medium and tomato juice.

This study was done to identify combinations of essential oils (EOs) that elicit synergistic antimicrobial effects against Leuconostoc citreum, a spoilage bacterium in vegetable and fruit juices. Twenty-four EOs were tested for antimicrobial activity against L. citreum using an agar well diffusion assay. Ten EOs showed relatively strong antimicrobial activity. Among those, cinnamon bark, oregano, and thyme thymol EOs showed the strongest activity (minimal inhibitory concentration = 1.25 µL/mL). It was confirmed that a combination of oregano and thyme thymol EOs had a synergistic antimicrobial activity (fractional inhibitory concentration index = 0.3750) using a checkerboard assay. This combination also had a synergistic antimicrobial activity against L. citreum in tomato juice. An initial population of L. citreum in tomato juice (5.3 log CFU/mL) increased to ≥ 8.6 log CFU/mL within 48 h at 15 °C and 24 h at 25 °C. Populations reached 6.8-7.2 log CFU/mL in tomato juice containing either 0.156 mL/mL oregano or thyme thymol. However, in the presence of both EOs (0.156 µL/mL in total), populations of L. citreum were significantly lower (5.7-6.5 log CFU/mL) (P ≤ 0.05). Findings will be useful when developing non-thermal food preservation technologies to increase the shelf-life of juices and other foods not containing synthetic preservatives.

Glove-mediated transfer of Listeria monocytogenes on fresh-cut cantaloupe.

The common use of gloves in retail practices represents a potential route for cross contamination of foodborne pathogens in fresh-cut produce. Using fresh-cut cantaloupe as a food model, we investigated factors that may influence glove-mediated cross contamination by Listeria monocytogenes and developed mathematical models to illustrate the patterns of transfer during fresh-cutting practices. Contact time (2, 5, 10 s), contact pressure (0.05, 0.18, 0.37 psi), and glove type (nitrile, polyvinyl chloride, polyethylene) did not have a significant effect on transfer of L. monocytogenes from cantaloupe rind to flesh, or from flesh to flesh. However, glove type appeared to affect L. monocytogenes transfer from the stem scar tissue to cantaloupe flesh (P = 0.0371). Transfer from rind pieces that had been washed with water was significantly higher than transfer from pieces that had not been washed (P = 0.0006). Predictive modeling and experimental validation suggested that transfer of L. monocytogenes on cantaloupe flesh persists over 85 pieces through consecutive contacts with a gloved hand. Findings of the study provide new scientific data to aid researchers, retailers, and caterers in safety risk assessments of fresh-cut practices used to prepare cantaloupes and other produce items.

Synergistic activities of gaseous oregano and thyme thymol essential oils against Listeria monocytogenes on surfaces of a laboratory medium and radish sprouts.

We investigated combinations of gaseous essential oils (EO gases) for their synergistic inhibitory activities against Listeria monocytogenes on a laboratory medium and radish sprouts. The minimum inhibitory concentrations and minimum lethal concentrations of oregano, thyme thymol, and cinnamon bark EO gases against L. monocytogenes were 0.0781 µL/mL on nutrient agar supplemented with glucose and bromocresol purple (NGBA). A checkerboard assay showed that combinations of oregano and thyme thymol EO gases and of oregano and cinnamon bark EO gases exert the strongest synergistic antilisterial activity (fractional inhibitory concentration index [FICI] = 0.3750). A combination of thyme thymol and cinnamon bark EO gases also had a synergistic effect (FICI = 0.5000) on L. monocytogenes on NGBA. Combinations of oregano and thyme thymol EO gases were tested for synergistic antimicrobial activity against L. monocytogenes on radish sprouts. A combination of these gases, each at 0.313 µL/mL, caused a significant (P ≤ 0.05) reduction in the number of L. monocytogenes on radish sprouts compared with reductions caused by treatment with oregano or thyme thymol EO gas alone at the same concentration. Our findings provide information that will be useful when developing antimicrobial applications using EO gases to control L. monocytogenes in the food industry.

Development of a decontamination method to inactivate Acidovorax citrulli on Cucurbitaceae seeds without loss of seed viability.

BACKGROUND: Acidovorax citrulli is a plant pathogen causing bacterial fruit blotch in Cucurbitaceae family. Applying high concentration of disinfectants to seeds containing plant pathogen may substantially decrease the germination rate of seeds. Therefore, it is necessary to develop a hurdle technology which can inactivate plant pathogens without decreasing seed viability. This study was conducted to develop a decontamination method to inactivate the plant pathogen Acidovorax citrulli on Cucurbitaceae seeds by sequential treatments with aqueous chlorine dioxide (ClO2 ), drying, and dry heat. RESULTS: The maximum ClO2 concentration that did not lower germination rates of cucumber, honeydew melon, and watermelon seeds was ca. 100 µg mL-1 of ClO2 for 5 min. Optimal incubation conditions for drying seeds that had been treated with aqueous ClO2 were determined as 25 °C and 43% relative humidity (RH) for 48 h. The maximum dry-heat temperature that did not reduce germination rates of seeds, which had been treated with ClO2 and dried at 25 °C, was 60 °C at 43% RH for 24 h. When seeds containing A. citrulli (6.4-7.0 log CFU g-1 ) were treated with aqueous ClO2 (50 µg mL-1 , 5 min), dried (25 °C, 43% RH, 24 h), and dry heated (60 °C, 43% RH, 24 h), the pathogen was inactivated to below the detection limit from all three seed types (<-0.5 log CFU g-1 ). CONCLUSION: The decontamination conditions to inactivate A. citrullii from Cucurbitaceae seeds without decreasing the seed viability were determined (sequential treatment with ClO2 [50 µg mL-1 , 5 min], dried [25 °C, 43% RH, 24 h], and dry heated [60 °C, 43% RH, 24 h]). The results of this study may also be applicable to other plant pathogens on other types of seeds. © 2019 Society of Chemical Industry.

Inhibition of Listeria monocytogenes using biofilms of non-pathogenic soil bacteria (Streptomyces spp.) on stainless steel under desiccated condition.

Of the 1648 microbial isolates from 133 soil samples collected from 30 diverse locations in the Republic of Korea, two isolates exhibited strong antilisterial activity and ability to grow to high populations (>8.0 log CFU/ml) in Bennett's broth. Isolates were identified as Streptomyces lactacystinicus (strain Samnamu 5-15) and Streptomyces purpureus (strain Chamnamu-sup 4-15). Both isolates formed biofilms on the surface of stainless steel coupons (SSCs) immersed in Bennett's broth within 24 h at 25 °C. Cells retained antilisterial activity after biofilm formation and showed significantly (P ≤ 0.05) enhanced resistance against dry conditions (43% relative humidity [RH]) compared to the cells not in biofilm. An initial population (ca. 3.2 log CFU/cm2) of Listeria monocytogenes inoculated on SSCs lacking Streptomyces biofilm decreased to 1.4 log CFU/cm2 within 48 h at 25 °C and 43% RH. In contrast, L. monocytogenes (3.3 log CFU/cm2) inoculated on SSCs containing Streptomyces biofilm decreased to populations below the theoretical detection limit (0.5 log CFU/cm2) within 24 h. The results indicate that biofilms formed by the Streptomyces spp. inhibitory to L. monocytogenes showed enhanced resistance to desiccation condition (43% RH) and effectively inhibited the growth of L. monocytogenes on the surfaces of SSCs. Antilisterial biofilms developed in this study may be applicable on desiccated environmental surfaces in food related environments such as food storage, handling, and processing facilities to enhance the microbiological safety of foods.

Synergistic antimicrobial activities of essential oil vapours against Penicillium corylophilum on a laboratory medium and beef jerky.

This study was done to determine the antifungal activities of essential oil (EO) vapours of 97 plants against Penicillium corylophilum and to test combinations of EO vapours for synergistic antifungal effects. Among 97 commercially available EOs extracted from plant parts, garlic, cinnamon bark, may chang (mountain pepper), citronella, thyme thymol, oregano, spearmint, and thyme linalool EO vapours exhibited relatively strong antifungal activities. The minimal inhibitory concentrations of these EO vapours were 0.0390-0.6250 µL/mL. A combination of cinnamon bark, citronella, and may chang EO vapours, as well as a combination of cinnamon bark and citronella EO vapours, showed synergistic inhibitory activities to P. corylophilum on a laboratory medium. A combination of cinnamon bark, citronella, and may chang EO vapours had synergistic activity in inhibiting growth of P. corylophilium on beef jerky. Observations reported here provide basic information valuable when developing strategies to inhibit the growth of P. corylophilum and possibly other moderately xerophilic molds on intermediate-moisture foods.

Inactivation of Salmonella Typhimurium on red chili peppers by treatment with gaseous chlorine dioxide followed by drying.

Chili pepper (Capsicum annuum L.) powder and other powdered spices containing chili peppers are occasionally contaminated with foodborne pathogens. We applied chlorine dioxide (ClO2) gas treatment to chili peppers prior to drying to inactivate Salmonella Typhimurium. Chili peppers inoculated with S. Typhimurium were exposed to ClO2 gas generated from 0.77 mL of aqueous ClO2 for 6 h at 25 °C and 100% relative humidity, followed by air drying at 55 °C for up to 24 h. Populations of S. Typhimurium and total aerobic bacteria (TAB) on peppers, chromaticity values, and moisture content were determined after various treatment times. S. Typhimurium (ca. 5.6 log cfu/g) was reduced to <0.7 log cfu/g of peppers treated with ClO2 gas for 0.5 h at 25 °C, but was detected by enrichment (>1 cfu/10.8 g) after treatment for 4 h. The pathogen was not detected after treatment with ClO2 gas for 6 h. S. Typhimurium did not decrease significantly (P > 0.05) on peppers exposed to air at 25 °C for 6 h, but rapidly decreased to 1.6 log cfu/g after subsequent drying for 6 h at 55 °C. The initial number of TAB (ca. 6.5 log cfu/g) decreased to 1.4 log cfu/g after treatment with ClO2 gas for 0.5 h and was less than the detection limit (0.7 log cfu/g) after 2 h. TAB counts were not reduced by treating peppers with air for 6 h, but decreased to <0.7 log cfu/g after subsequent drying for 24 h at 55 °C. The lightness (L value) of treated and untreated chili peppers did not change during drying, but redness (a value) and yellowness (b value) decreased. Results showed that treatment of chili peppers containing S. Typhimurium (5.6 log cfu/g) with ClO2 gas for 6 h prior to drying at 55 °C reduced the population to < 1 cfu/10.8 g.

Antimicrobial activities of gaseous essential oils against Listeria monocytogenes on a laboratory medium and radish sprouts.

The aim of this study was to evaluate the antimicrobial activities of gaseous essential oils (EO gases) against Listeria monocytogenes on the surfaces of a laboratory medium and radish sprouts. We determined the minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) values of EO gases from eight EOs extracted from basil leaves, carrot seed, cinnamon bark, cinnamon leaves, clove flower buds, oregano leaves, thyme flowers (linalool), and thyme leaves (thymol) against L. monocytogenes on a nutrient agar supplemented with 1% glucose and 0.025% bromocresol purple (NGBA). Oregano, thyme thymol, and cinnamon bark EO gases showed the strongest antilisterial activities (MIC and MLC, 78.1µL/L). We also investigated the inhibitory and lethal activities of these gases against L. monocytogenes on the surface of radish sprouts. The number of L. monocytogenes after exposure to EO gases at ≥156µL/L was significantly (P≤0.05) lower than that of untreated L. monocytogenes. For example, the initial number of L. monocytogenes on the surface of radish sprouts (ca. 6.3logCFU/g) decreased by 1.4logCFU/g within 24h at 30°C and 43% relative humidity (RH) without EO gas treatment, whereas the number of L. monocytogenes after exposure to oregano, thyme thymol, and cinnamon bark EO gases at 156µL/L decreased by 2.1, 2.1, and 1.8logCFU/g, respectively, after 24h. Although EO gases exerted greater lethal activities at higher concentrations (312 and 625µL/L), L. monocytogenes on the surface of radish sprouts was not completely inactivated. The number of L. monocytogenes on sprouts treated with oregano, thyme thymol, and cinnamon bark EO gases at 625µL/L decreased by 2.7-3.0logCFU/g after 24h at 30°C and 43% RH. Results indicate that EO gases that showed antilisterial activities on a laboratory medium also exhibited reduced lethal activity on the surface of radish sprouts. These findings will be useful when developing strategies to inactivate L. monocytogenes and possibly other foodborne pathogens on sprouts and perhaps other foods using EO gases.

Development of non-pathogenic bacterial biofilms on the surface of stainless steel which are inhibitory to Salmonella enterica.

Non-pathogenic bacterial biofilms were developed on the surface of stainless steel possessing desiccation tolerance and antimicrobial activity against Salmonella enterica. Three bacteria exhibiting strong antimicrobial activities against S. enterica were isolated from various soils, foods, and food-contact surfaces. Isolates were identified as Pseudomonas extremorientalis (strain Lettuce-28), Paenibacillus peoriae (strain Lettuce-7), and Streptomyces cirratus (strain Geumsan-207). These bacteria grew rapidly and formed biofilms within 24 h on the surface of stainless steel coupons (SSCs) immersed in laboratory media (tryptic soy broth or Bennet's broth) at 25 °C. Cells in biofilms had enhanced tolerance to desiccation (exposure to 43% atmospheric relative humidity [RH]) and retained antimicrobial activity against S. enterica. Populations of S. enterica deposited on SSCs containing biofilm formed by Ps. extremorientalis strain Lettuce-28, for example, decreased by > 2.5 log CFU/coupon within 24 h at 25 °C and 43% RH, while the number of cells inoculated on SSCs lacking biofilm decreased by 1.5 log CFU/coupon. Antimicrobial activities of the three antagonistic bacteria against S. enterica persisted in desiccated biofilms. This study provides insights to developing strategies to inactivate Salmonella and perhaps other foodborne pathogens on abiotic surfaces using non-pathogenic antagonistic bacteria.

Retention of Viability of Salmonella in Sucrose as Affected by Type of Inoculum, Water Activity, and Storage Temperature.

Outbreaks of salmonellosis have been associated with consumption of high-sugar, low-water activity (aw) foods. The study reported here was focused on determining the effect of storage temperature (5 and 25°C) on survival of initially high and low levels of Salmonella in dry-inoculated sucrose (aw 0.26 ± 0.01 to 0.54 ± 0.01) and wet-inoculated sucrose (aw 0.24 ± 0.01 to 0.44 ± 0.04) over a 52-week period. With the exception of dry-inoculated sucrose at aw 0.26, Salmonella survived for 52 weeks in dry- and wet-inoculated sucrose stored at 5 and 25°C. Retention of viability was clearly favored in sucrose stored at 5°C compared with 25°C, regardless of level or type of inoculum or aw. Survival at 5°C was not affected by aw. Initial high-inoculum counts of 5.18 and 5.25 log CFU/g of dry-inoculated sucrose (aw 0.26 and 0.54, respectively) stored for 52 weeks at 5°C decreased by 0.56 and 0.53 log CFU/g; counts decreased by >4.18 and >4.25 log CFU/g in samples stored at 25°C. Inactivation rates in wet-inoculated sucrose were similar to those in dry-inoculated sucrose; however, a trend toward higher persistence of Salmonella in dry- versus wet-inoculated sucrose suggests there was a higher proportion of cells in the wet inoculum with low tolerance to osmotic stress. Survival patterns were similar in sucrose initially containing a low level of Salmonella (2.26 to 2.91 log CFU/g). The pathogen was recovered from low-inoculated sucrose stored at 5°C for 52 weeks regardless of type of inoculum or aw and from dry-inoculated sucrose (aw 0.54) and wet-inoculated sucrose (aw 0.24) stored at 25°C for 12 and 26 weeks, respectively. Results emphasize the importance of preventing contamination of sucrose intended for use as an ingredient in foods not subjected to a treatment that would be lethal to Salmonella.

Inactivation of Salmonella enterica in chicken feces on the surface of eggshells by simultaneous treatments with gaseous chlorine dioxide and mild wet heat.

The aim of this study was to investigate the lethal effects of simultaneous treatments with gaseous chlorine dioxide (ClO2) and mild wet heat (55 °C at 100% relative humidity [RH]) on Salmonella enterica in chicken feces on the surface of eggshells. Gaseous ClO2 production decreased significantly (P ≤ 0.05) as the RH (23, 43, 68, 85, and 100%) at 25 °C was increased. The lethality of gaseous ClO2 against S. enterica in feces on eggshells increased significantly (P ≤ 0.05) as RH increased. For example, when treated with gaseous ClO2 at 85 and 100% RH at 25 °C, S. enterica (5.9 log CFU/egg) was inactivated within 4 h. In contrast, at 23, 43, and 68% RH, the pathogen remained at 5.1, 5.0, and 2.8 log CFU/egg, respectively, after 6 h. Finally, when eggshells surface-contaminated with S. enterica (5.8 log CFU/egg) were treated with gaseous ClO2 (peak concentration of ClO2: 185.6 ppm) at 100% RH and 55 °C, inactivation occurred within 1 h. These results indicate that treatment of surface-contaminated shell eggs with gaseous ClO2 at elevated RH and temperature is effective in inactivating S. enterica. These observations will be useful when developing an effective sanitation program to enhance the microbiological safety of shell eggs.

Inhibition of Staphylococcus aureus by antimicrobial biofilms formed by competitive exclusion microorganisms on stainless steel.

The goal of this study was to develop a desiccation resistant antimicrobial surface using biofilm of competitive exclusion (CE) microorganism inhibitory to Staphylococcus aureus. We isolated 161 microorganisms from soils, foods, and food-contact surfaces that are inhibitory to S. aureus. Among them, three CE microorganisms (Streptomyces spororaveus strain Gaeunsan-18, Bacillus safensis strain Chamnamu-sup 5-25, and Pseudomonas azotoformans strain Lettuce-9) exhibiting strong antibacterial activity and high growth rates were selected for evaluation. These isolates formed biofilms within 24h on stainless steel coupons (SSCs) immersed in Bennet's broth and tryptic soy broth at 25°C. Cells in these biofilms showed significantly (P≤0.05) enhanced resistance to a desiccation (43% relative humidity [RH]) compared to those attached to SSCs but not in biofilms. The antimicrobial activities of biofilms formed by these isolates on SSCs against S. aureus at 25°C and 43% RH were determined. Compared to SSCs lacking biofilms formed by CE microorganisms, populations of S. aureus on SSCs harboring CE biofilms were significantly lower (P≤0.05). Results indicate that persistent antimicrobial activity against S. aureus on stainless steel surfaces can be achieved by the presence of biofilms of CE microorganisms. This information will be useful when developing strategies to improve the microbiological safety of foods during storage, processing, and distribution by facilitating the development of effective antimicrobial food-contact surfaces.

Sanitizing radish seeds by simultaneous treatments with gaseous chlorine dioxide, high relative humidity, and mild heat.

Sanitizing radish seeds intended for edible sprout production was achieved by applying simultaneous treatments with gaseous chlorine dioxide (ClO2), high relative humidity (RH, 100%), and mild heat (55°C). Gaseous ClO2 was produced from aqueous ClO2 (0.66ml) by mixing sulfuric acid (5% w/v) with sodium chlorite (10 mg/mL) in a sealed container (1.8L). Greater amounts of gaseous ClO2 were measured at 23% RH (144ppm after 6h) than at 100% RH (66ppm after 6h); however, the lethal activity of gaseous ClO2 against naturally occurring mesophilic aerobic bacteria (MAB) on radish seeds was significantly enhanced at 100% RH. For example, when exposed to gaseous ClO2 at 23% RH, the number of MAB on radish seeds decreased from 3.7logCFU/g to 2.6logCFU/g after 6h. However, when exposed to gaseous ClO2 at 100% RH for 6h, the MAB population decreased to 0.7logCFU/g after 6h. Gaseous ClO2 was produced in higher amounts at 55°C than at 25°C, but decreased more rapidly over time at 55°C than at 25°C. The lethal activity of gaseous ClO2 against MAB on radish seeds was greater at 55°C than at 25°C. When radish seeds were treated with gaseous ClO2 (peak concentration: 195ppm) at 100% RH and 55°C, MAB were reduced to populations below the detectable level (<-0.7logCFU/g) within 2h without decreasing the seed germination rate (97.7%). The lethality of combined treatments against artificially inoculated Escherichia coli O157:H7 was also evaluated. When exposed to gaseous ClO2 at 100% RH and 55°C for 6h, the initial number of E. coli O157:H7 (3.5logCFU/g) on radish seeds decreased to below the detection limit (0.7logCFU/g) by direct plating but it was not eliminated from seeds. The germination rate of radish seeds was not significantly (P>0.05) decreased after treatment for 6h. The information reported here will be useful when developing decontamination strategies for producing microbiologically safe radish seed sprouts.

Comparison of New and Traditional Culture-Dependent Media for Enumerating Foodborne Yeasts and Molds.

Fifty-six foods and food ingredients were analyzed for populations of naturally occurring yeasts and molds using Petrifilm rapid yeast and mold (RYM) count plates, Petrifilm yeast and mold (YM) count plates, dichloran rose bengal chloramphenicol (DRBC) agar plates, acidified potato dextrose agar (APDA) plates, and dichloran 18% glycerol (DG18) agar plates. Colonies were counted after incubating plates for 48, 72, and 120 h at 25°C. Of 56 foods in which either yeasts or molds were detected on at least one medium incubated for 120 h, neither yeasts nor molds were detected in 55.4, 73.2, 21.4, 19.6, and 71.4% of foods plated on the five respective media and incubated for 48 h; 10.7, 14.3, 3.6, 1.8, and 19.6% of foods were negative after 72 h, and 3.6, 1.8, 0, 0, and 0% of foods were negative after 120 h. Considering all enumeration media, correlation coefficients were 0.03 to 0.97 at 48 h of incubation; these values increased to 0.75 to 0.99 at 120 h. Coefficients of variation for total yeasts and molds were as high as 30.0, 30.8, and 27.2% at 48, 72, and 120 h, respectively. The general order of performance was DRBC = APDA > RYM Petrifilm > YM Petrifilm ≥ DG18 when plates were incubated for 48 h, DRBC > APDA > RYM Petrifilm > YM Petrifilm ≥ DG18 when plates were incubated for 72 h, and DRBC > APDA > RYM Petrifilm > YM Petrifilm > DG18 when plates were incubated for 120 h. Differences in performance among media are attributed to the diversity of yeasts and molds likely to be present in test foods and differences in nutrient, pH, and water activity requirements for resuscitation of stressed cells and colony development.