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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.

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.

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.

Nuts, including almonds, are occasionally contaminated with Salmonella spp. In this study, we used chlorine dioxide (ClO

The objectives of this study were to screen essential oil (EO) vapors for inhibitory activities against Staphylococcus aureus on a laboratory medium and to validate their antibacterial effects on dried black peppercorns (Piper nigrum). Among 84 commercially available EOs, using a vapor disc assay, vapors from caraway seed, carrot seed, cinnamon bark, citronella, kanuka, lemongrass, may chang, oregano, and thyme thymol EOs were found to be more inhibitory to S. aureus. The minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) of nine EO vapors from these plant materials were measured on a laboratory medium. Citronella, lemongrass, and oregano EO vapors had the lowest MIC and MLC values, at 0.0781 and 0.1563 μL/mL, respectively. Using a checkerboard assay, partial synergism in inhibitory activity of a combination of citronella and lemongrass EO vapors was observed (fractional inhibitory concentration index = 0.6250). When tested against S. aureus on black peppercorns, citronella, lemongrass, and oregano EO vapors, both individually and in combination, showed a significant antibacterial effect (P ≤ 0.05). Notably, a combination of citronella and lemongrass EO vapors (1:4 vol ratio) showed significant synergistic activity, reducing the S. aureus population by ca. ≥ 1.3–1.8 log CFU/g more than treatment with single EOs (P ≤ 0.05). The results of this study provide important basic information for developing technologies to control S. aureus on the surfaces of dried foods using EO vapors.

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.

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, 78.1 μL/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 ≥ 156 μL/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. 6.3 log CFU/g) decreased by 1.4 log CFU/g within 24 h 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 156 μL/L decreased by 2.1, 2.1, and 1.8 log CFU/g, respectively, after 24 h. Although EO gases exerted greater lethal activities at higher concentrations (312 and 625 μL/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 625 μL/L decreased by 2.7–3.0 log CFU/g after 24 h 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.

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.

Nuts, including almonds, are occasionally contaminated with Salmonella spp. In this study, we used chlorine dioxide (ClO2) gas to inactivate S. enterica subsp. Enterica serovar Enteritidis on almonds. Almonds inoculated with a single strain of S. Enteritidis (8.95 log cfu/mL) were exposed to ClO2 gas generated from 1.0 or 1.5 mL ClO2 solution in a sealed container at 50 or 60 °C (43% relative humidity) for up to 10 h. The concentration of ClO2 gas peaked at 354–510 and 750–786 ppm within 0.5 h upon deposition of 1.0 and 1.5 mL of aqueous ClO2, respectively, and gradually decreased thereafter. Population of S. Enteritidis on almonds treated at 50 °C decreased to 1.70–2.32 log cfu/sample within 1 h of exposure to ClO2 gas and decreased to below the detection limit (1.7 log cfu/sample) at all ClO2 concentrations after 8 h. At 60 °C, the microbial population fell below the detection limit within 1 h, regardless of the volume of ClO2 solution supplied. Microbial survival on almonds treated with ClO2 gas and stored at 12 or 25 °C was observed for up to 8 weeks and the organism was not recovered from the almonds treated for 10 h and stored at 12 °C for 2–8 weeks. The lightness (L value) and redness (a value) of almonds treated for 10 h were not changed by ClO2 gas treatment, but yellowness (b value) increased. Results showed that Salmonella on almonds was successfully inactivated by ClO2 gas treatment and the microbial survival did not occur during storage.

The objective of this study was to identify combinations of plant essential oils (EOs) that have synergistic antilisterial effects in raw organic tomato juice. Among the EOs approved for addition to organic foods in the Republic of Korea, twenty-four EOs were screened for antilisterial activity using a well diffusion assay. Six EOs (cinnamon bark, lemon, lemongrass, oregano, peppermint, and thyme thymol) showed relatively strong inhibitory activities (diameter of zone of inhibition ≥ 35.0 mm) against Listeria monocytogenes. The minimal inhibitory concentrations (MICs) and minimal lethal concentrations (MLCs) of these EOs against L. monocytogenes in a laboratory medium were determined. Oregano and thyme thymol EOs had the lowest MIC (0.3125 μL/mL), and cinnamon bark, oregano, and thyme thymol EOs had the lowest MLC (0.6250 μL/mL). The synergistic antilisterial activities of combinations of EOs in a laboratory medium were determined using a modified checkerboard assay. Combinations of cinnamon bark and lemongrass EOs, cinnamon bark and thyme thymol EOs, and lemongrass and thyme thymol EOs showed synergistic antilisterial activities (fractional inhibitory concentration index = 0.5). A combination of cinnamon bark and thyme thymol EOs was tested for synergistic antilisterial activity in tomato juice incubated at 10 °C and 25 °C for up to 48 h. The number of L. monocytogenes in juice treated with combined EOs at a total concentration of 0.3125 μL/mL decreased significantly (P ≤ 0.05) compared to numbers in juice treated with 0.3125 μL/mL of either cinnamon bark EO or thyme thymol EO alone. These observations show the potential for using combinations of EOs for controlling L. monocytogenes in juice released from cut or bruised raw organic tomatoes intended for consumption in food service or home settings.

Halloysite nanotubes (HNTs), which are natural nanomaterials, have a hollow tubular structure with about 15 nm inner and 50 nm outer diameters. Because of their tubular shape, HNTs loaded with various materials have been investigated as functional nanocapsules. In this study, thyme essential oil (TO) was encapsulated successfully in HNTs using vacuum pulling methods, followed by end-capping or a layer-by-layer surface coating process for complete encapsulation. Nanocapsules loaded with TO were mixed with flexographic ink and coated on a paper for applications as food packaging materials. Scanning electron microscopy and transmission electron microscopy were used to characterize the morphology of the nanocapsules and to confirm the TO loading of the nanocapsules. Fourier transform infrared spectroscopy and thermogravimetric analyses analysis were used to complement the structural information. In addition, the controlled release of TO from the nanocapsules showed sustained release properties over a period of many days. The results reveal that the release properties of TO in these nanocapsules could be controlled by surface modifications such as end-capping and/or surface coating of bare nanocapsules. The packaging paper with TO-loaded HNT capsules was effective in eliminating against Escherichia coli during the first 5 d and showed strong antibacterial activity for about 10 d.

An effective bactericidal cold-marinating method for beef products is described, exploiting the synergism between soy sauce and natural compounds (carvacrol, CV or thymol, TM) to reduce microbiological risks. Beef slices inoculated with Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium (3.1-3.5logCFU/g) were marinated in a teriyaki sauce with or without CV and TM (0.3 and 0.5%). After 1, 3, and 7days at 4°C, indigenous microflora population, color, lipid oxidation, marinade uptake, and pH of marinated beef and leftover marinade samples were examined. Teriyaki sauce alone did not reduce or inhibit any of the target pathogens or indigenous bacteria, while 0.5% CV- or TM-containing teriyaki sauce inactivated all inocula without recovery within 7days (p0.05). The pathogens relocated from the beef into the leftover marinade (3.0-3.4logCFU/mL) were also completely inactivated. The treatment inhibited growth of indigenous aerobic bacteria (p0.05) and inactivated coliform bacteria. Physicochemical parameters were not significantly affected (p0.05).

The effect of a combined treatment using aqueous chlorine dioxide (ClO2 ) and hot-air drying to inactivate Bacillus cereus spores on red chili peppers was evaluated. Ten washed and dried pepper samples, each comprising half of a single pepper (Capsicum annuum L.), were inoculated with B. cereus spore suspension. The inoculated samples were washed with sodium hypochlorite (NaOCl; 50, 100, or 200 μg/mL) or ClO2 (50, 100, or 200 μg/mL) solution for 1 min and then air-dried (25 ± 1 °C, 47 ± 1% relative humidity), which was followed by drying with hot air at 55 °C for up to 48 h. The spore populations on the samples were enumerated and their aw and chromaticity values were measured. The spore numbers immediately after treatment with NaOCl and ClO2 were not significantly different. A more rapid reduction in spore numbers was observed in the samples treated with ClO2 than those treated with NaOCl during drying. A combined treatment of ClO2 and hot-air drying significantly reduced the spore populations to below the detection limit (1.7 log CFU/sample). B. cereus spores on chili peppers were successfully inactivated by washing with ClO2 solution followed by hot-air drying whereas the pepper color was maintained.

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 ( 0.05) decreased after treatment for 6h. The information reported here will be useful when developing decontamination strategies for producing microbiologically safe radish seed sprouts.

Germination rates of 11 types of sprout seeds (alfalfa, broccoli, kohlrabi, kyona, mustard, pak choi, red kohlrabi, red radish, red young radish, tatsoi, and violet radish) treated with ClO 2 (200 μg/ml, 5 min) or dry-heat (80 °C/23% relative humidity [RH], 24 h) were determined. Pak choi, red radish, and tatsoi seeds showed highest tolerance to both ClO 2 and dry-heat treatments. Next, pak choi seeds were inoculated with Escherichia coli O157:H7 (5.4 log CFU/g) or Salmonella enterica (4.8 log CFU/g) and sequentially treated with ClO 2 (200 μg/ml, 5 min), drying (45 °C/23% RH, 24 h), and dry heat (80 °C/23% RH, 48 h). E. coli O157:H7 was inactivated, but S. enterica was not eliminated (>3.8 log CFU/g reduction). Pak choi seeds inoculated with the pathogens were treated with ClO 2 , drying, and dry heat and subsequently sprouted for 5 days. When seeds were not completely decontaminated, initial populations of E. coli O157:H7 and S. enterica on seeds ( 5.3 and > 8.4 log CFU/g of sprouts, respectively. This study shows that sequential treatments of pak choi seeds with ClO 2 , drying, and dry heat are effective in reducing large numbers of E. coli O157:H7 and S. enterica without loss of seed viability.

There is a growing interest in the use of naturally-occurring antimicrobial agents such as plant essential oils (EOs) to inhibit the growth of hazardous and spoilage microorganisms in foods. Gaseous EOs (EO gases) have many potential applications in the food industry, including use as antimicrobial agents in food packaging materials and sanitizing agents for foods and food-contact surfaces, and in food processing environments. Despite the potentially beneficial applications of EO gases, there is no standard method to evaluate their antimicrobial activities. Thus, the present study was aimed at developing an experimental apparatus and protocol to determine the minimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) of EO gases against microorganisms. A sealed experimental apparatus was constructed for simultaneous evaluation of antimicrobial activities of EO gases at different concentrations without creating concentration gradients. A differential medium was then evaluated in which a color change allowed for the determination of growth of glucose-fermenting microorganisms. Lastly, an experimental protocol for the assessment of MIC and MLC values of EO gases was developed, and these values were determined for 31 EO gases against Escherichia coli O157:H7 as a model bacterium. Results showed that cinnamon bark EO gas had the lowest MIC (0.0391 μl/ml), followed by thyme-thymol EO gas (0.0781 μl/ml), oregano EO gas (0.3125 μl/ml), peppermint EO gas (0.6250 μl/ml), and thyme-linalool EO gas (0.6250 μl/ml). The order of the MLC values of the EO gases against the E. coli O157:H7 was thyme-thymol (0.0781 μl/ml)

This study focused on yuba films coated with active agents to improve microbiological and oxidative stability and the feasibility of preserving beef patties. Yuba films (YC) coated with oregano essential oil (YO), grapefruit seed extract (YG), and their mixture (YOG) were characterized and applied to beef patties. YO (2.56 × 10 − 10 g m/Pa·sec· m 2 ) displayed exceptional water barrier properties compared to YC (4.27 × 10 − 10 g m/Pa·sec· m 2 ) . All coated yuba films showed increased antioxidant capacity, particularly YG. In disc diffusion, all coated yuba films (inhibition zones ≥ 20.0 mm) were strongly effective against L. monocytogenes, whereas YO (27.17 mm) and YOG (29.00 mm) were effective against E. coli O157:H7. In the application to beef patties, YG beef patties (1.69 MDA mg/kg) showed the highest oxidative stability as a result of lipid oxidation inhibition (YC 1.88 MDA mg/kg). Furthermore, YG beef patties maintained the highest quality of meat color, which was dark purplish-red, during storage. While E. coli O157:H7 in beef patties was significantly inhibited by YO and YOG, the growth of L. monocytogenes in beef patties was impeded by all coated yuba films throughout the storage period. Overall, YOG can be potentially used to increase microbiological safety and extend the shelf life of beef patties.

The aim of this study was to use chlorine dioxide (ClO2) gas to inactivate Aspergillus flavus on green coffee beans. Green coffee beans inoculated with A. flavus were exposed to ClO2 gas generated from 0.75, 1, 1.25, or 1.5 mL of ClO2 solution in a sealed container at 25 or 50 °C (100% relative humidity [RH]) for up to 10 h. Numbers of A. flavus on beans treated at 25 °C decreased by 1.1–2.2 log cfu/bean within 1 h of exposure to ClO2 and decreased to below the detection limit (≤1.0 log cfu/bean) at all ClO2 concentrations after 10 h. At 50 °C, the microbial population reached the detection limit within 0.5 h regardless of the ClO2 solution concentration. Beans exposed to gas for 10 h at 25 °C were stored for 14 days under conditions of 43, 75, or 100% RH and 4, 12, or 25 °C. At 4 °C, visible mold growth was not established regardless of RH. After 12 days, mold was observed only at 25 °C. At 100% RH, beans stored under the same conditions without ClO2 gas treatment showed mold formation at 13 and 4 days at 12 and 25 °C, respectively.

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.

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 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

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.

We produced a Pseudomonas veronii biofilm on the surface of a stainless steel that is inhibitory to Escherichia coli O157:H7. Pseudomonas veronii strain KACC 81051BP, isolated from lettuce, readily formed biofilm on the surface of stainless steel coupons (SSCs) immersed in tryptic soy broth at 25°C. Cells showed significantly (P ≤ 0·05) enhanced tolerance to desiccation stress (43% relative humidity (RH)) and retained antimicrobial activity against E. coli O157:H7. The number of E. coli O157:H7 (control; 4·1 ± 0·1 log CFU per coupon) on sterile SSCs decreased to 2·7 ± 0·2 log CFU per coupon after exposure to 43% RH at 25°C for 48 h, while the population of E. coli O157:H7 (4·1 ± 0·0 log CFU per coupon) on SSCs containing P. veronii biofilm decreased to below the theoretical detection limit (1·5 log CFU per coupon) within 24 h. The antimicrobial biofilm produced on stainless steel may have application in preventing cross-contamination by E. coli O157:H7 on other abiotic surfaces in food-contact environments. Significance and impact of the study The presence of Escherichia coli O157:H7 on environmental surfaces of food manufacturing, transportation and storage facilities is a significant food safety concern because it can result in cross-contamination of food products. In this study, we developed a Pseudomonas veronii biofilm on the surface of a stainless steel that inhibits the growth of E. coli O157:H7. Since P. veronii in biofilm resists desiccation, it provides persistent antimicrobial activity. Information presented here provides novel and practical insights to developing biological strategies to inactivate E. coli O157:H7 on diverse surfaces in food processing and handling environments.

We compared the microbiological quality of chicken eggshells obtained from a traditional wholesale market and a modern supermarket. We also determined the survival and growth characteristics of naturally occurring mesophilic aerobic bacteria (MAB) and artificially inoculated Salmonella enterica on eggshells under various environmental conditions (presence of chicken feces, temperature [4, 12, or 25 °C], and relative humidity [RH; 43 or 85%]). The populations of MAB, coliforms, and molds and yeasts on eggshells purchased from a traditional wholesale market were significantly (P ≤ 0.05) higher than those from a modern supermarket. In the second study, when we stored uninoculated eggs under various storage conditions, the population of MAB on eggshells (4.7-4.9 log CFU/egg) remained constant for 21 days, regardless of storage conditions. However, when eggshells were inoculated with S. enterica and stored under the same conditions, populations of the pathogen decreased significantly (P ≤ 0.05) under all tested conditions. Survival of S. enterica increased significantly (P ≤ 0.05) in the presence of feces, at low temperatures, and at low RH. These observations will be of value when predicting the behavior of microorganisms on eggshells and selecting storage conditions that reduce the populations of S. enterica on eggshells during distribution.

We evaluated the lethal activity of gaseous chlorine dioxide (ClO2) against Bacillus cereus spores attached to and in biofilm formed on a stainless steel surface. Aqueous ClO2 was prepared by mixing sulfuric acid (5% w/v) with sodium chlorite (10mg/mL), and gaseous ClO2 was produced by vaporization of aqueous ClO2 in an air-tight container. The concentration of gaseous ClO2 in the air within the container increased rapidly at first but gradually decreased over time. The lethality of gaseous ClO2 against B. cereus spores attached to stainless steel coupons (SSCs) and in biofilm formed by the pathogen on SSCs was determined. The B. cereus spores attached to SSCs (5.3±0.1logCFU/coupon) were completely inactivated within 1h at 25°C when treated with gaseous ClO2 (peak concentration: 115.3±5.0 parts per million [ppm]). The total number of vegetative cells and spores in biofilm formed by B. cereus on SSCs was 5.9±0.3logCFU/coupon; the spore count was 5.3±0.1logCFU/coupon. The vegetative cells and spores in biofilm were completely inactivated within 6h (peak concentration: 115.3±5.0ppm). Results show that B. cereus spores in biofilms are more resistant to gaseous ClO2 than are attached spores not in biofilms. Gaseous ClO2 was, nevertheless, very effective in killing B. cereus spores in biofilm on the surface of stainless steel. Results show promise for application of gaseous ClO2 to enhance the microbiological safety of foods that may come in contact with stainless steel and possibly other hard surfaces on which B. cereus biofilms have formed.

Food-grade galactooligosaccharide (GOS) with low water activity (aw of ca. 0.7) is used as an ingredient in various foods. We evaluated heat tolerances of Salmonella, Cronobacter sakazakii, and Pediococcus acidilactici at temperatures (70 to 85°C) used during the saturation process of GOS by comparing decimal reduction time (D-values) and thermal resistance constants (z-values). To determine the D- and z-values, GOS containing Salmonella (5.1 to 5.8 log CFU/g) or C. sakazakii (5.3 to 5.9 log CFU/g) was heat treated at 70, 77.5, or 85°C for up to 40, 25, or 15 s, respectively, and GOS containing P. acidilactici (6.1 to 6.5 log CFU/g) was heat treated at 70, 77.5, or 85°C for up to 150, 75, or 40 s, respectively. The D-values were calculated using a linear model for heating time versus microbial population for each bacterium. When the D-values for Salmonella, C. sakazakii, and P. acidilactici in GOS were compared, the thermal resistance of all bacteria decreased as the temperature increased. Among the three bacteria, P. acidilactici had higher D-values than did Salmonella and C. sakazakii. The z-values of Salmonella, C. sakazakii, and P. acidilactici were 30.10, 33.18, and 13.04°C, respectively. Overall order of thermal resistance was P. acidilactici > Salmonella ≈ C. sakazakii. These results will be useful for selecting appropriate heat treatment conditions for the decontamination of pathogenic microorganisms during GOS manufacturing.

Halloysite nanotubes (HNTs), which are natural nanomaterials, have a hollow tubular structure with about 15 nm inner and 50 nm outer diameters. Because of their tubular shape, HNTs loaded with various materials have been investigated as functional nanocapsules. In this study, thyme essential oil (TO) was encapsulated successfully in HNTs using vacuum pulling methods, followed by end-capping or a layer-by-layer surface coating process for complete encapsulation. Nanocapsules loaded with TO were mixed with flexographic ink and coated on a paper for applications as food packaging materials. Scanning electron microscopy and transmission electron microscopy were used to characterize the morphology of the nanocapsules and to confirm the TO loading of the nanocapsules. Fourier transform infrared spectroscopy and thermogravimetric analyses analysis were used to complement the structural information. In addition, the controlled release of TO from the nanocapsules showed sustained release properties over a period of many days. The results reveal that the release properties of TO in these nanocapsules could be controlled by surface modifications such as end-capping and/or surface coating of bare nanocapsules. The packaging paper with TO-loaded HNT capsules was effective in eliminating against Escherichia coli during the first 5 d and showed strong antibacterial activity for about 10 d.

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.

Fresh produce is usually eaten raw without cooking or heating, which may increase the probability of foodborne infection. The microbiological quality of 11 types of fresh, raw vegetables (romaine lettuce, sesame leaves, crown daisy, garlic chives, iceberg lettuce, cabbage, broccoli, leek, chili pepper, capsicum, and zucchini) purchased at retail markets in Iksan, Korea as affected by cultivation method (environmentally friendly vegetables [organic, pesticide-free, and low-pesticide vegetables] and conventionally grown vegetables) and harvest season was determined. Escherichia coli O157:H7 and Salmonella were not detected in all samples of vegetables tested. Aerobic mesophiles (>6 log cfu/g) were detected in environmentally friendly romaine lettuce and crown daisy and environmentally friendly and conventionally grown garlic chives, which also contained coliforms (>3 log cfu/g). Sesame leaf and crown daisy (regardless of cultivation method), as well as conventionally grown romaine lettuce and leek, contained >1 log cfu/g of E. coli. The overall microbiological quality of environmentally friendly and conventionally grown vegetables was not significantly different (P > 0.05). However, there were seasonal effects on populations of coliforms and generic E. coli on vegetables. The greatest numbers of microorganisms were isolated from environmentally friendly or conventionally grown vegetables purchased in winter. The vegetables, regardless of cultivation method or season, should be subjected to appropriate antimicrobial treatment to enhance their microbial safety.

We studied the survival and growth patterns of Bacillus cereus, mesophilic aerobic bacteria (MAB), and molds and yeasts (MY) on rough and milled brown and white Korean rice stored at 12 and 21 °C and 43, 68, and 85% relative humidity (RH) for up to 24 wk. The initial populations of MAB present on rough rice, brown rice, and white rice were 7.7, 5.7, and 3.3 log CFU/g, respectively, and remained constant or decreased (P ≤ 0.05) by 0.7-1.8 log CFU/g during storage. The initial populations of B. cereus on the three types of laboratory-inoculated rice were 3.1-3.8 log CFU/g and remained constant (P > 0.05) during storage, regardless of degree of milling, storage temperature, and RH. The initial populations of MY on rough rice, brown rice, and white rice were 6.2, 4.2, and 2.1 log CFU/g, respectively. At 12 °C and 85% RH, the MY increased significantly (P ≤ 0.05) only on brown rice; however, at 21 °C and 85% RH, MY increased (P ≤ 0.05) on all types of rice during storage. These observations will be useful when assessing conditions affecting survival of B. cereus and determining environmental conditions necessary to prevent growth of potentially mycotoxigenic molds on various types of milled rice during storage.

Food-grade galactooligosaccharide (GOS) has a low water activity (aw). We investigated the survival of Escherichia coli, Salmonella spp., Pseudomonas aeruginosa, Listeria monocytogenes, Staphylococcus aureus, Aspergillus flavus and Saccharomyces cerevisiae in GOS as affected by inoculum levels and storage temperatures. The five bacteria (inoculated at 3 or 6 log cfu/g), and the fungus A. flavus (3 log cfu/g) and S. cerevisiae (3 log cfu/g), were inoculated into GOS and the material stored at 4, 22 or 50C for 28 days. When ca. 6 log cfu/g of microorganisms were inoculated into GOS, the final numbers of L. monocytogenes and S. aureus (thus after 28 days of storage) were higher than those of E. coli, Salmonella spp. and P. aeruginosa, at all storage temperatures tested. In GOS inoculated with ca. 3 log cfu/g of microorganisms, A. flavus numbers were the highest after 28 days. S. cerevisiae levels were lower than those of gram-positive but higher than those of gram-negative bacteria. The overall level of survival by temperature was 4C > 22C > 50C, regardless of inoculum level. Thus, microbial survival in low-aw GOS was affected by temperature and inoculum level but the microorganisms did not grow during the storage. Practical Applications This study showed that survival of microorganisms in GOS was affected by storage temperature and inoculum level. Microorganisms did not grow in GOS at all temperatures tested, but they still survived in GOS for up to 28 days upon storage temperature. This information will be useful in development of effective microbiological safety strategies to be incorporated in preparation or storage of low-aw food and food ingredients.

The effect of a combined treatment using aqueous chlorine dioxide (ClO

Hyun-Sun Seo, Seonhwa Kim, Jinsol Kim, Jaejoon Han, and Jee-Hoon Ryu*Department of Food Bioscience and Technology, College of Life Sciences and Biotechnology, Korea UniversityAbstract We investigated the delay of kimchi fermentation by the addition of plant extracts. Fifteen plant extracts werescreened for inhibitory activity aginst Lactobacillus plantarum by using an agar well diffusion assay, and determined theminimal inhibitory concentration (MIC) and minimal lethal concentration (MLC) were determined. The lowest MIC forgrapefruit seed extract (GFSE; 0.0313 mg/mL) was determined, followed by Caesalpinia sappan L. extract (CSLE; 0.25 mg/mL), and oregano essential oil (OREO; 1.0 mg/mL). GFSE, CSLE, and OREO were individually added to kimchi, andincubated the samples at 10 for up to 20 days. Results showed that the addition of GFSE (0.3 and 0.5%), CSLE (0.1, 0.3,and 0.5%), or OREO (0.5 and 1.0%) led to a significant increase in the pH of kimchi, and also a significant reduction in thenumbers of lactic acid bacteria. Taken together, the addition of natural antimicrobial agents can delay kimchi fermentation.Keywords: kimchi, fermentation, plant extract

The objective of this study was to control the survival or biofilm formation of Cronobacter spp. on stainless steel surfaces using Paenibacillus polymyxa. The antibacterial activity of a cell-free culture supernatant (CFCS) of P. polymyxa against Cronobacter spp. was found to vary with P. polymyxa incubation time. Maximum activity occurred when P. polymyxa was incubated at 25 or 30 °C for 96 h. When the CFCS was introduced to Cronobacter spp. adhered to stainless steel strips at 25 °C for up to 72 h, the CFCS successfully inhibited Cronobacter biofilm formation. Additionally, stainless steel surfaces with a preformed P. polymyxa biofilm were exposed to Cronobacter spp. suspensions in PBS or 0.1% peptone water at 3, 5, or 7 log CFU/mL to facilitate its attachment. The Cronobacter population significantly decreased on this surface, regardless of inoculum level or carrier, when the P. polymyxa biofilm was present. However, the microbial population decreased within 6 h and remained unchanged thereafter when the surface was immersed in an inoculum suspended in 0.1% peptone water at 5 or 7 log CFU/mL. These results indicate that P. polymyxa is able to use a promising candidate competitive-exclusion microorganism to control Cronobacter spp.

This study evaluated the inactivation of Cronobacter spp. on radish seeds using ClO2 treatment followed by storage in modified atmosphere (MA) for up to 4 days. Radish seeds were inoculated with Cronobacter spp. (6 log cfu/g), adapted to nalidixic acid and treated with a ClO2 aqueous solution (0, 50 or 100 μL/mL). The seeds were then stored in an ambient air or MA (10% O2, 10% CO2 and 80% N2; 5% O2, 10% CO2 and 85% N2; or 10% O2, 0% CO2 and 90% N2) for up to 4 days. The Cronobacter spp. (6.2 log cfu/g) populations on the radish seeds were reduced to 3.9, 3.8 or 2.6 log cfu/g after treatment with 0, 50 or 100 μL/mL ClO2, respectively, and then to 3.7, 3.1 or ≤ 1.6 log cfu/g after 2 h of drying. After 2–3 days of storage under MA (5 or 10% O2 with 10% CO2), no Cronobacter spp. were detected on the seeds treated with 100 μL/mL aqueous ClO2. More than 94.3% of the radish seeds germinated after ClO2 treatment followed by exposure to an MA, regardless of the ClO2 concentration. Practical Applications These results indicate that sprout seeds should be treated with sanitizers at an appropriate concentration. Sanitizer treatment of the seeds is more critical than following storage conditions for enhancing microbial safety. In addition, sprout seeds should be dried or stored after ClO2 treatment to increase its antimicrobial activity because a greater reduction in microbial populations occurred when the seeds treated with ClO2 were dried or stored than immediately after the treatment.

We developed a DNA microarray that contains random genomic DNA fragments of Listeria monocytogenes, validated its diagnostic abilities using cells grown in laboratory media and milk, and established enrichment conditions for detection of a low population of L. monocytogenes in milk. Genomic DNA of L. monocytogenes strain ATCC 19111 was fractionated by agarose gel electrophoresis after being cleaved using several different pairs of restriction enzymes. Sixty DNA fragments of different sizes were randomly selected and spotted onto an amine-coated glass slide. To validate diagnostic ability, probes on the DNA microarray were hybridized with genomic DNA extracted from L. monocytogenes, other Listeria spp., and foodborne pathogenic bacteria belonging to other genera grown in laboratory media. The DNA microarray showed 98-100% positive hybridization signals for the 16 strains of L. monocytogenes tested, 7-85% positive signals for 9 strains of other Listeria spp., and 0-32% positive signals for 13 strains of other types of foodborne pathogens. In milk, the detection limit of the DNA microarray was approximately 8 log CFU/mL. When milk contained L. monocytogenes (3-4 log CFU/mL) with other types of bacteria (Bacillus spp., B. cereus, Salmonella Montevideo, Peudomonas aeruginosa, and Yersinia enterocolitica; ca. 3 log CFU/mL each), L. monocytogenes enriched in UVM modified Listeria enrichment broth at 37°C for 24h was successfully detected by the DNA microarray. Results indicate that the DNA microarray can detect L. monocytogenes and distinguish it from other Listeria spp. and other foodborne pathogens in laboratory media and milk. This platform will be useful when developing a DNA microarray to rapidly and simultaneously detect and identify various foodborne pathogens in foods.

A sachet releasing allyl isothiocyanate (AIT) vapor was developed and its effectiveness as an antimicrobial packaging system for fresh spinach was evaluated. AIT was encapsulated in calcium alginate beads (AIT beads) and the release of AIT as affected by temperature and relative humidity (RH) was determined. The release rate of AIT from beads increased with increased temperature, but was not significantly affected by RH. A low-density polyethylene (LDPE) sachet containing AIT beads (AIT sachet) was prepared and the rate of release of AIT as affected by film thickness and temperature was studied. The release of AIT from sachets increased significantly with decreased LDPE thickness and increased temperature. Finally, antimicrobial effects of the AIT sachet against Escherichia coli O157:H7 and molds and yeasts (MY) on fresh spinach leaves were determined. E. coli O157:H7 and MY had similar sensitivities to AIT vapor. The number of E. coli O157:H7 on spinach leaves (5.6logCFU/leaf) decreased by 1.6-2.6logCFU/leaf at 4°C and 2.1-5.7logCFU/leaf at 25°C within 5days. The reduction of E. coli O157:H7 was significantly greater at 85% RH than at 43% RH. Reductions on control leaves (no AIT sachet) did not exceed 1.2logCFU/leaf under any of the test conditions. Results of the study will be useful when developing antimicrobial packaging systems to increase the microbiological safety and extend the shelf life of spinach and possibly other fresh produces.

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.

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.

Aims: To determine survival and colonization of Escherichia coli O157:H7 on spinach leaves as affected by inoculum level and carrier, temperature and relative humidity (r.h.). Methods and Results: Spinach leaves were inoculated with suspensions of E. coli O157:H7 in distilled water (DW) and 0·1% peptone water (PW) and incubated at 4, 12 and 25°C and 43, 85 and 100% r.h. The number of E. coli O157:H7 on leaves (5·6 or 1·9 log CFU per leaf) inoculated using DW as a carrier medium increased significantly at 25°C and 100% r.h. within 120 h but remained constant or decreased significantly under other test conditions. E. coli O157:H7 on leaves (5·4 log CFU per leaf) inoculated using PW as a carrier increased significantly within 72 and 24 h, respectively, at 12 or 25°C and 100% r.h.; counts using a low inoculum (2·2 log CFU per leaf) increased significantly within 24 h at 25°C. Conclusions: Escherichia coli O157:H7 can colonize on spinach leaves at 12 or 25°C in a 100% r.h. environment. Organic matter in the inoculum carrier may provide protection and nutrients which enhance survival and colonization. Significance and Impact of the Study: Colonization of E. coli O157:H7 on spinach leaves as affected by organic matter in the inoculum, temperature and r.h. was determined. These observations will be useful when developing strategies to prevent growth of E. coli O157:H7 on pre- and postharvest spinach.

A sour kimchi product with an elevated amount of γ-aminobutyric acid (GABA) was produced using starter lactic acid bacteria (LAB) for mukeunjee kimchi fermentation. The starter LAB were screened and isolated from the commercial mukeunjee kimchi product that showed the highest GABA content and was identified as Lactobacillus buchneri. The maximum GABA production of L. buchneri in MRS media was 5.83 mg/mL at pH 4.2 and the addition of 3% NaCl did not significantly (p>0.05) changed GABA production of L. buchneri. The amount of GABA in L. buchneri-inoculated sour kimchi was 61.65 mg/100 g, which represented about 8 times higher than the L. buchneri uninoculated kimchi (control kimchi). A sensory evaluation test of L. buchneri-inoculated sour kimchi and control kimchi showed that the L. buchneri inoculation may produce more sour kimchi than control kimchi, but there was no difference in the general acceptance between the 2 kimchi products (p>0.05).

We determined the combined effectiveness of ClO(2) (200 and 500 μg/ml, 5 min), air drying [25 °C, 40% relative humidity (RH), 2 h], and mild dry heat (55 °C, 23% RH, up to 48 h) treatments in killing total aerobic bacteria (TAB), Escherichia coli O157:H7, and molds and yeasts (MY) on radish seeds. A 5.1-log reduction in the number of TAB was achieved on radish seeds treated with 200 or 500 μg/ml ClO(2) followed by air drying for 2 h and dry heat treatment for 48 h or 24 h, respectively. When radish seeds were treated with 200 and 500 μg/ml ClO(2), air dried, and heat treated for 12 h and 6 h, respectively, the initial population of E. coli O157:H7 (5.6 log CFU/g) on seeds was reduced to an undetectable level (0.8 log CFU/g). However, the pathogen was detected in 5-day-old sprouts. The reduction of MY (1.2-1.0 log CFU/g) on radish seeds under similar experimental conditions was not changed significantly during subsequent heat treatment up to 48 h. Results show that treating radish seeds with 500 μg/ml ClO(2), followed by air dried at 25 °C for 2 h and heat treatment at 55 °C for 36 h achieved a5-log CFU/g reduction of TAB and E. coli O157:H7. These observations will be useful when developing effective strategies and practices to enhance the microbiological safety of radish sprouts.

In this study, we present the successful detection of food-borne pathogens using randomly selected non-sequenced genomic DNA probes-based DNA microarray chips. Three food-borne pathogens, Staphylococcus aureus, Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium), and Bacillus cereus, were subjected for the preparation of the DNA microarray probes. Initially, about 50 DNA probes selected randomly from non-sequenced genomic DNA of each pathogen were prepared by using a set of restriction enzyme pairs. The proto-type of DNA microarray chip for detecting three different pathogens simultaneously was fabricated by using those DNA probes prepared for each pathogen. This proto-type DNA microarray has been tested with three target pathogens and additional seven bacteria, and successfully verified with a few cross-hybridized probes. After this primary verification of the DNA microarray hybridization, this proto-type DNA microarray chip was redesigned and successfully optimized by eliminating a few cross-hybridized probes. The specificity of this redesigned DNA microarray chip to each pathogen was confirmed without any serious cross-hybridizations, and its multiplexing capability in its pathogen detection was found to be possible. This randomly selected non-sequenced genomic DNA probes-based DNA microarray was successfully proved to be the high-throughput simultaneous detection chip for the detection of food-borne pathogens, without knowing the exact sequence information of the target bacteria. This could be the first fabrication of DNA microarray chip for the simultaneous detection of different kinds of food-borne pathogens.

Aims: To fabricate a DNA chip containing random fragments of genomic DNA of Yersinia enterocolitica and to verify its diagnostic ability. Methods and Results: A DNA microarray chip was fabricated using randomly fragmented DNA of Y. enterocolitica. Chips were hybridized with genomic DNA extracted from other Y. enterocolitica strains, other Yersinia spp. and bacteria in different genera. Genomic DNA extracted from Y. enterocolitica showed a significantly higher hybridization rate compared with DNA of other Yersinia spp. or bacterial genera, thereby distinguishing it from other bacteria. Conclusions: A DNA chip containing randomly fragmented genomic DNA from Y. enterocolitica can detect Y. enterocolitica and clearly distinguish it from other Yersinia spp. and bacteria in different genera. Significance and Impact of the Study: A microarray chip containing randomly fragmented genomic DNA of Y. enterocolitica was fabricated without sequence information, and its diagnostic ability to identify Y. enterocolitica was verified.