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This study examined the disinfection efficacy and mechanism of electrolyzed water (EW) on Bacillus cereus biofilms. B. cereus strains, ATCC 14579 and Korean Collection for Type Cultures (KCTC) 13153 biofilms, were formed on stainless steel (SS) and plastic slide (PS) coupons. Mature biofilms were treated with slightly acidic EW (SAEW), acidic EW (AEW), and basic EW (BEW). SAEW (available chlorine concentration, 25 ± 1.31 mg L-1; pH 5.71 ± 0.16; and oxidation reduction potential, 818 to 855 mV) reduced ATCC 14579 biofilms on plastic slides to below the detection limit within 30 s. However, biofilms on SS coupons showed a higher resistance to the SAEW treatment. When the disinfection activities of three types of EW on biofilms were compared, AEW showed a higher bactericidal activity, followed by SAEW and BEW. In contrast, BEW showed a significantly ( P < 0.05) higher biofilm dispersal activity than AEW and SAEW. SAEW disinfection of the B. cereus biofilms was due to the disruption of the B. cereus plasma membrane. The higher resistance of biofilms formed on the SS coupon might be due to the higher number of attached cells and extracellular polymeric substances formation that reacts with the active chlorine ions, such as hypochlorous acid and hypochlorite ion of SAEW, which decreased the disinfection efficacy of SAEW. This study showed that the EW treatment effectively disinfected B. cereus biofilms, providing insight into the potential use of EW in the food processing industry to control the biofilm formation of B. cereus.

This study investigated the antigenotoxic effects of enzymatic browning reaction products (PEBRPs) obtained by reaction of polyphenol compounds with oxidase extracted from potato. Each of the PEBRPs by themselves at 100 mg/kg did not induce an increased frequency of micronucleated polychromatic erythrocytes (MNPCEs) irrespective of the sampling time (up to 72 h), while the treatment with benzo[a]pyrene (B[a]P) significantly increased the incidence of MNPCEs (P < 0.05). Significant reductions were observed in the frequencies of MNPCEs (P < 0.05) when all PEBRPs were given to the mice 12 h before they were exposed to 100 mg/kg of B[a]P and inhibitory effects were 60%, 70%, and 60% in the catechol (Ca)-PEBRPs, hydroxyhydroquinone (HHQ)-PEBRPs, and pyrogallol (Py)-PEBRPs, respectively. When three kinds of PEBRPs were fed to mice 12 h before injecting 100 mg/kg of B[a]P, the most significant decrease (P < 0.05) in the frequencies of MNPCEs induced by B[a]P were observed and the relative frequency inhibitions b...

Individual and combined antimicrobial effects of monolaurin and heat on planktonic, 1-day adherent, or 7-day adherent cells of Listeria monocytogenes were determined to evaluate biofilm removal from stainless steel. Planktonic cells were more sensitive to heat and monolaurin than were cells attached to stainless steel. Young (1-day) biofilm cells were more sensitive to each treatment than were old (7-day) biofilm cells. Adherent cells were destroyed by 50 μg/ml monolaurin combined with heating at 65°C for 5 min. Cells in a rich nutrient environment were more resistant to treatment than were cells in a depleted nutrient environment. Results demonstrate the usefulness of combining chemical and physical treatments to control L. monocytogenes biofilm problems in the food industry.

Microbiological analyses were made on samples of cooked blue crab taken immediately after debacking and either forced-air cooling or static-air cooling. Forced-air cooling had significantly lower (P

The aim of this study was to model the growth kinetics of Listeria monocytogenes on ready-to-eat ham and sausage at different temperatures (4 to 35°C). The observed data fitted well with four primary models (Baranyi, modified Gompertz, logistic, and Huang) with high coefficients of determination (R(2) > 0.98) at all measured temperatures. After the mean square error (0.009 to 0.051), bias factors (0.99 to1.06), and accuracy factors (1.01 to 1.09) were obtained in all models, the square root and the natural logarithm model were employed to describe the relation between temperature and specific growth rate (SGR) and lag time (LT) derived from the primary models. These models were validated against the independent data observed from additional experiments using the acceptable prediction zone method and the proportion of the standard error of prediction. All secondary models based on each of the four primary models were acceptable to describe the growth of the pathogen in the two samples. The validation results indicate that the optimal primary model for estimating the SGR was the Baranyi model, and the optimal primary model for estimating LT was the logistic model in ready-to-eat (RTE) ham. The Baranyi model was also the optimal model to estimate the SGR and LT in RTE sausage. These results could be used to standardize predictive models, which are commonly used to identify critical control points in hazard analysis and critical control point systems or for the quantitative microbial risk assessment to improve the food safety of RTE meat products.

The combined effects of ultrasonication and slight acidic electrolyzed water were investigated to improve the microbial safety of brown rice against Bacillus cereus infection and to evaluate the growth kinetics of these bacteria during storage of untreated and treated rice at various temperatures (5, 10, 15, 20, 25, 30, and 35°C). The results indicate that this combination treatment was bactericidal against B. cereus, resulting in an approximately 3.29-log reduction. Although B. cereus can be efficiently reduced by treatment, temperature abuse during storage can allow B. cereus to recover and grow. A primary growth model (Baranyi and Roberts equation) was fitted to the raw growth data from untreated (control) and treated samples to estimate growth rate, lag time, and maximum population density, with a low standard error of the residuals (≤0.140) and high adjusted coefficient of determination (>0.990). The growth curves obtained from the Baranyi and Roberts model indicated that B. cereus grew more slowly on treated brown rice than on untreated brown rice. Secondary models predicting the square root of the maximum growth rate and the natural logarithm of the lag time as a function of temperature were satisfactory (bias factor = 0.993 to 1.013; accuracy factor = 1.290 to 1.352; standard error of prediction = 18.828 to 36.615%). Inactivation results and the model developed and validated in this study provided reliable and valuable growth kinetics information for quantitative microbiological risk assessment studies of B. cereus on brown rice.

Foods grown in organic production systems have been described as representing an increased risk to public health compared with foods from conventional production. Leafy vegetables (spinach, romaine lettuce, and green sesame leaves) grown in organic and conventional systems were collected from various areas in Korea and examined using standard culture methods to compare the microbiological quality of the produce grown in the two agricultural systems. The 354 samples of these leafy vegetables were analyzed for levels of indicator bacteria (aerobic bacteria, coliforms, and Escherichia coli) and the prevalence of the pathogens Staphylococcus aureus, E. coli O157:H7, Listeria monocytogenes, Bacillus cereus, and Salmonella. Aerobic bacteria and coliforms were detected in all vegetable types, but nonpathogenic E. coli was below the limit of detection in all samples. B. cereus was the most prevalent pathogen, found on 7 (11.1%) of the 63 organic spinach samples. The prevalence of S. aureus was highest in organic sesame leaves; it was found on 5 (8.0%) of the 63 samples. The prevalence of L. monocytogenes was highest on organic romaine lettuce and spinach; it was found in 4 (6.4%) of 63 samples of each type of vegetable. E. coli O157:H7 found on only 1 (1.58%) of 55 conventional spinach samples. These results suggest that farming type at most only slightly affects the hygienic quality of leafy vegetables, and no effect was found for sample collection area. Salmonella was not isolated from any of the conventional or organic leafy vegetables. These results do not support the hypothesis that organic produce poses a substantially greater risk of pathogen contamination than does conventional produce.

The detection ability and selectivity of Karmali agar was improved by supplementation of an extended-spectrum β-lactamase inhibitor, potassium clavulanate. The optimum concentration of potassium clavulanate (0.5 μg/ml) in Karmali agar was determined by inoculation of 50 Campylobacter and 30 extended-spectrum β-lactamase-producing E. coli strains onto normal and modified Karmali agar containing various concentrations of the agent. Eighty retail carcasses were rinsed with 400 ml of buffered peptone water. The rinse samples were enriched in 2 × blood-free Bolton enrichment broth at 42°C for 48 h and then were streaked onto normal and modified Karmali agar containing 0.5 μg/ml potassium clavulanate. The suspicious colonies were subcultured on Columbia blood agar and confirmed by colony PCR. In chicken carcass samples, the modified Karmali agar showed a significantly greater isolation rate than normal Karmali agar (42.5 versus 21.3%; P0.05). Furthermore, the selectivity of the modified Karmali agar was also significantly higher (P0.05) than that of the normal Karmali agar, as seen by comparison of the number of contaminated agar plates (83.8 versus 97.5%) and the growth index (1.67 versus 2.91) of the non-Campylobacter colonies.

The objective of this study was to investigate the microbial contamination levels (aerobic bacteria plate count [APC], coliforms, Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes) in mixed pressed ham and cooked sausage. A total of 180 samples were collected from factories with and without hazard analysis critical control point (HACCP) systems at four steps: after chopping (AC), after mixing (AM), cooling after the first heating process, and cooling after the second heating process. For ham, APCs and coliform and E. coli counts increased when ingredients were added to the meat at the AC step. Final product APC was 1.63 to 1.85 log CFU/g, and coliforms and E. coli were not detected. S. aureus and L. monocytogenes were found in nine (15.0%) and six (10.0%) samples, respectively, but only at the AC and AM steps and not in the final product. Sausage results were similar to those for ham. The final product APC was 1.52 to 3.85 log CFU/g, and coliforms and E. coli were not detected. S. aureus and L. monocytogenes were found in 29 (24.2%) and 25 (20.8%) samples at the AC and AM steps, respectively, but not in the final product. These results indicate that the temperature and time of the first and second heating are of extreme importance to ensure the microbiological safety of the final product regardless of whether a HACCP system is in place. Microorganism contamination must be monitored regularly and regulations regarding sanitization during processing should be improved. Education regarding employee personal hygiene, environmental hygiene, prevention of cross-contamination, ingredient control, and step-by-step process control is needed to reduce the risk of food poisoning.

This study was conducted to investigate the disinfection efficacy of hurdle treatments (thermosonication plus slightly acidic electrolyzed water [SAcEW]) and to develop a model for describing the effect of storage temperatures (4, 10, 15, 20, 25, 30, and 35°C) on the growth of Escherichia coli O157:H7 on fresh-cut kale treated with or without (control) thermosonication combined with SAcEW. The hurdle treatments of thermosonication plus SAcEW had strong bactericidal effects against E. coli O157:H7 on kale, with approximately 3.3-log reductions. A modified Gompertz model was used to describe growth parameters such as specific growth rate (SGR) and lag time (LT) as a function of storage temperature, with high coefficients of determination (R(2)0.98). SGR increased and LT declined with rising temperatures in all samples. A significant difference was found between the SGR values obtained from treated and untreated samples. Secondary models were established for SGR and LT to evaluate the effects of storage temperature on the growth kinetics of E. coli O157:H7 in treated and untreated kale. Statistical evaluation was carried out to validate the performance of the developed models, based on the additional experimental data not used for the model development. The validation step indicated that the overall predictions were inside the acceptable prediction zone and had lower standard errors, indicating that this new growth model can be used to assess the risk of E. coli O157:H7 contamination on kale.

Three important foodborne pathogens, Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus, are of great concern for food safety. They may also coexist in food matrices and, in the case of B. cereus and S. aureus, the resulting illnesses can resemble each other owing to similar symptoms. Therefore, their simultaneous detection may have advantages in terms of cost savings and rapidity. Given this context, a rapid multiplex real-time PCR high-resolution melt curve assay for the simultaneous detection of these three pathogens in food was developed. The assay successfully detected B. cereus (gyrB), L. monocytogenes (hly), and S. aureus (nuc) in a single reaction, and the average melting temperatures were 76.23, 80.19, and 74.01°C, respectively. The application of SYTO9 dye and a slow melt curve analysis ramp rate (0.1°C/s) enabled the production of sharp, high-resolution melt curve peaks that were easily distinguishable from each other. The detection limit in food (milk, rice, and lettuce) was 3.7 × 103 CFU/g without an enrichment step and 3.7 × 101 CFU/g following the 10-h enrichment. Hence, the assay developed here is specific and sensitive, providing an efficient tool for implementation in food for the simultaneous detection of B. cereus, L. monocytogenes, and S. aureus.

Recent Staphylococcus aureus outbreaks linked to meat and poultry products underscore the importance of understanding the growth kinetics of S. aureus in these products at different temperatures. Raw pork, ham, and sausage (each 10 ± 0.3 g) were inoculated with a three-strain cocktail of S. aureus, resulting in an initial level of ca. 3 log CFU/g. Samples were stored isothermally at 10, 15, 20, 25, 30, 35, and 40°C, and S. aureus was enumerated at appropriate time intervals. The square root model was developed using experimental data collected from S. aureus grown on all samples (where data from raw pork, ham, and sausage were combined) so as to describe the growth rate of S. aureus as a function of temperature. The model was then compared with models for S. aureus growth on each individual sample in the experiments (raw pork, ham, or sausage) and the S. aureus ComBase models, as well as models for the growth of different types of pathogens (S. aureus, Escherichia coli O157:H7, Clostridium perfringens, Salmonella serovars, and Salmonella Typhimurium) on various types of meat and poultry products. The results show that the S. aureus model developed here based on the pooled data from all three pork products seems suitable for the prediction of S. aureus growth on different pork products under isothermal conditions from 10 to 25°C, as well as for S. aureus growth on different meat and poultry products at higher temperatures between 20 and 35°C. Regardless of some high deviations observed at temperatures between 25 and 40°C, the developed model still seems suitable to predict the growth of other pathogens on different types of meat and poultry products over the temperature ranges used here, especially for E. coli O157:H7 and Salmonella Typhimurium. The developed model, therefore, may be useful for estimating the effects of storage temperature on the behavior of pathogens in different meat and poultry products and for microbial risk assessments evaluating meat safety.

Bacillus cereus was divided into emetic toxin (cereulide)- and enterotoxin-producing strains, but emetic toxin-producing B. cereus is difficult to detect immunochemically. Screening methods for emetic toxin-producing B. cereus are needed. The objectives of this study were to identify and detect emetic toxin-producing B. cereus among 160 B. cereus strains, and to compare enterotoxin production and phenotypic characteristics between the emetic toxin-producing and enterotoxin-producing strains. Forty emetic toxin-producing B. cereus strains were determined with high-pressure liquid chromatography-mass spectrometry analysis. Among the emetic toxin-producing strains (n = 40), 31 (77.5%) and 3 (7.5%) strains produced nonhemolytic enterotoxin (NHE) and hemolysin BL (HBL) enterotoxins, respectively. In addition, 107 (89.2%) and 100 (83.3%) strains produced NHE and HBL enterotoxins among the enterotoxin-producing strains (n = 120). The number of strains positive for starch hydrolysis, salicin fermentation, and hemolysis among the emetic toxin-producing strains were 3 (7.5%), 3 (7.5%), and 26 (65.0%), respectively, and among enterotoxin-producing strains, these numbers were 101 (84.2%), 100 (83.3%), and 111 (92.5%), respectively. In particular, the three emetic toxin-producing B. cereus strains (JNHE 6, JNHE 36, and KNIH 28) produced the HBL and NHE enterotoxins and were capable of starch hydrolysis and salicin fermentation. The absence of HBL enterotoxin and certain phenotypic properties, such as starch hydrolysis and salicin fermentation, indicates that these properties were not critical characteristics of the emetic toxin-producing B. cereus tested in this study.

This study was conducted to develop a predictive model to estimate the growth of Listeria monocytogenes on fresh pork during storage at constant temperatures (5, 10, 15, 20, 25, 30, and 35°C). The Baranyi model was fitted to growth data (log CFU per gram) to calculate the specific growth rate (SGR) and lag time (LT) with a high coefficient of determination (R(2)0.98). As expected, SGR increased with a decline in LT with rising temperatures in all samples. Secondary models were then developed to describe the variation of SGR and LT as a function of temperature. Subsequently, the developed models were validated with additional independent growth data collected at 7, 17, 27, and 37°C and from published reports using proportion of relative errors and proportion of standard error of prediction. The proportion of relative errors of the SGR and LT models developed herein were 0.79 and 0.18, respectively. In addition, the standard error of prediction values of the SGR and LT of L. monocytogenes ranged from 25.7 to 33.1% and from 44.92 to 58.44%, respectively. These results suggest that the model developed in this study was capable of predicting the growth of L. monocytogenes under various isothermal conditions.

A total of 410 domestic Korean food samples were analyzed for the presence of Listeria spp. by the conventional U.S. Department of Agriculture protocol, and presumptive strains were identified by morphological, cultural and biochemical tests according to Bergey's manual and confirmed by API-Listeria kit. Among the total 410 food samples, 46 samples (11.2%) were found to be contaminated with Listeria species. Among the 46 strains of Listeria spp. isolates, 8 strains (17.42%) for Listeria monocytogenes, 3 strains (6.5%) for Listeria seeligeri, 33 strains (71.7%) for Listeria innocua, and 2 strains (4.4%) for Listeria welshimeri were identified, respectively. Also, only beef, chicken, pork, frozen foods, and sausage were contaminated with L. monocytogenes, and the other products were free of L. monocytogenes. Of 46 Listeria spp. isolates, L. innocua (71.7%) was the most predominantly isolated in a variety of foods compared to other Listeria spp. An in vitro virulence assay for Listeria spp. using myeloma and hybridoma cells from murine and human sources was performed. The result showed that only L. monocytogenes killed approximately 95 to 100% hybridoma cells after 6 h and the other Listeria species, such as L. innocua, L. seeligeri, and L. welshimeri strains had about 0 to 10% lethal effect on hybridoma cells. Also, an antibiotic susceptibility test showed that Listeria spp. isolates were very susceptible to the antibiotics tested, except for nalidixic acid. Also, serotyping results showed 75% of L. monocytogenes isolates from beef, chicken, and frozen pizza belonged to serotype 1 and 25% from sausage were type 4.

Bacillus cereus is frequently isolated from a variety of foods, including vegetables, dairy products, meats, and other raw and processed foods. The bacterium is capable of producing an enterotoxin and emetic toxin that can cause severe nausea, vomiting, and diarrhea. The objectives of this study were to assess and model the probability of enterotoxin production of B. cereus in a broth model as affected by the broth pH and storage temperature. A three-strain mixture of B. cereus was inoculated in tryptic soy broth adjusted to pH 5.0, 6.0, 7.2, 8.0, and 8.5, and the samples were stored at 15, 20, 25, 30, and 35°C for 24 h. A total of 25 combinations of pH and temperature, each with 10 samples, were tested. The presence of enterotoxin in broth was assayed using a commercial test kit. The probabilities of positive enterotoxin production in 25 treatments were fitted with a logistic regression to develop a probability model to describe the probability of toxin production as a function of pH and temperature. The resulting model showed that the probabilities of enterotoxin production of B. cereus in broth increased as the temperature increased and/or as the broth pH approached 7.0. The model described the experimental data satisfactorily and identified the boundary of pH and temperature for the production of enterotoxin. The model could provide information for assessing the food poisoning risk associated with enterotoxins of B. cereus and for the selection of product pH and storage temperature for foods to reduce the hazards associated with B. cereus.

The effect of temperature, pH, and monolaurin on the growth and survival of Listeria monocytogenes in tryptic soy broth with yeast extract (TSBYE) was investigated. TSBYE medium containing 5, 6, 7, 8, or 9 μg/ml monolaurin was adjusted to pH 5.0, 5.5, or 7.0, inoculated with L. monocytogenes , and incubated at 7, 15, or 35°C. The effect of sublethal concentrations of monolaurin was measured by calculating generation times. Results indicated that temperature and pH were very important factors influencing the efficacy of monolaurin against L. monocytogenes . Bactericidal effects were observed at pH 5.0 but not observed at pH 5.5 or pH 7.0. Further, lethal effects of monolaurin increased as temperature increased at constant pH. Conversely, bacteriostatic effects on growth increased as temperature decreased at constant pH. At constant temperature the bactericidal and bacteriostatic effects of monolaurin increased as the pH of the medium decreased. Generation time of the organism as significantly (P < 0.05) influenced by pH, temperature, concentration of monolaurin, or their combined interactions. Thus, the results indicate that pH and temperature interact to affect the antimicrobial potential of monolaurin against L. monocytogenes .

Incidence and properties of Bacillus cereus strains naturally present in cereals were evaluated by phenotypic characterization, antibiotic susceptibility testing, and pulsed-field gel electrophoresis. Of 293 cereal samples tested, 73 (25%) contained B. cereus strains. Incidence of B. cereus isolates varied with respect to sample; they were found in 15 (37%) of 83 brown rice samples, 23 (37%) of 63 glutinous rice samples, 16 (21%) of 76 barley samples, and 19 (27%) of 71 Job's tears samples. All B. cereus isolates from cereals were positive for diarrheal toxin genes. The isolates were susceptible to most of the antibiotics tested, but they were highly resistant to ampicillin, cefepime, oxacillin, and penicillin. Of the genes assayed by the PCR technique, a high frequency of nheA (99%) and hblDC (84%) was found in the genomic DNA of cereal-associated isolates, whereas cytK was less common (55%). From the strains carrying the hblDC genes, 93% produced enterotoxin HBL. B. cereus isolates did not have significant genetic homology. The genetic diversity and toxic potential differ among the strains isolated from cereals. These results provide important information on toxin gene profiles of cereal-associated B. cereus for population studies.

A number of studies on the influence of acid on Escherichia coli O157:H7 have shown considerable strain differences, but limited information has been reported to compare the acid resistance based on the different sources of E. coli O157:H7 isolates. The purpose of this study was to determine the survival of E. coli O157:H7 strains isolated from five sources (foods, bovine carcasses, bovine feces, water, and human) in 400 mM acetic acid solutions under conditions that are typical of acidified foods. The isolates from bovine carcasses, feces, and water survived acetic acid treatment at pH 3.3 and 30 degrees C significantly (Por = 0.05) better than did any food or human isolates. However, resistance to acetic acid significantly increased as temperature decreased to 15 degrees C for a given pH, with little (Por = 0.05) difference among the different isolation sources. All groups of E. coli O157:H7 strains showed more than 1.8- to 4.5-log reduction at pH 3.3 and 30 degrees C after 25 min. Significantly reduced (less than 1-log reduction) lethality for all E. coli O157:H7 strain mixtures was observed when pH increased to 3.7 or 4.3, with little difference in acetic acid resistance among the groups. The addition of glutamate to the acetic acid solution or anaerobic incubation provided the best protection compared with the above conditions for all groups of isolates. These results suggest that temperature, pH, and atmospheric conditions are key factors in establishing strategies for improving the safety of acidified foods.

The risk of Staphylococcus aureus in ready-to-eat kimbab (rice rolled in laver) sold in Korea was evaluated by a mathematical modeling approach. Four nodes were constructed from preparation at retail to consumption. A predictive microbial growth model and survey data were combined with probabilistic modeling to simulate the level of S. aureus in a single kimbab at the time of consumption. We estimated the mean level of S. aureus to be 2.92 log CFU/g for a typical kimbab (150 to 200 g each) at the time of consumption. Our model also showed that 29.73% of the kimbabs hador = 100,000 S. aureus CFU/g, which poses some risk of illness, since some level of enterotoxin would be expected from toxigenic strains. However, because of the lack of dose-response models for staphylococcal enterotoxin, the final level of S. aureus in the kimbabs could not be used to estimate how many people would become ill from eating them. Correlation sensitivity results showed that consumer eating patterns and initial contamination levels at retail stores were the most significant risk factors for illness and that temperature control under 10 degrees C was a critical control point in kimbab retail establishments to prevent the growth of S. aureus.

A total of 673 ready-to-eat vegetable samples were collected in Korea from 2001 to 2002 and analyzed for the presence of Yersinia spp. We analyzed biotypes, serotypes, and susceptibility to 12 antibiotics and tested for virulence genes of pathogenic Yersinia enterocolitica isolates by PCR assay. Among the samples, 27 (4.0%) were found to be contaminated with Yersinia spp. Among the 27 strains of Yersinia spp. isolates, 18 strains (66.7%) of Y. enterocolitica, 5 strains (18.5%) of Y. frederiksenii, 3 strains (11.1%) of Y. intermedia, and 1 strain (3.7%) of Y. kristensenii were identified. According to the serotypes of Y. enterocolitica isolates, O:3 (11.1%) and O:5 (11.1%) were the most predominant, followed by O:8 (5.6%) and others (72.2%). For biotypes of Y. enterocolitica isolates, 1A (77.8%) was the most predominant, followed by 3B (11.1%), 3 (5.6%), and 5A (5.6%). Also, an antibiotic susceptibility test showed that Y. enterocolitica isolates were very susceptible to the antibiotics tested but highly resistant to ampicillin (94%), cephalothin (100%), and carbenicillin (83%). PCR assays with specific primers derived from yst and ail genes of Y. enterocolitica were applied to confirm the presence of pathogenic Y. enterocolitica. Among the 18 strains of Y. enterocolitica isolates, only 3 strains (O:3/1A, UT/3B, and UT/1A isolated from Chinese cabbage, onion, and spinach, respectively) were shown to have a virulence gene.

Individual and combined antimicrobial effects of monolaurin and acetic acid on Listeria monocytogenes planktonic cells or stainless-steel-adherent cells were determined in order to evaluate cell viability during a 25-min exposure period at 25 degrees C. A 10(7)-colony-forming units (CFU)/ml population of planktonic cells was completely inactivated by the synergistic combination of 1% acetic acid with 50 or 100 microg/ml of monolaurin within 25 or 20 min, respectively. Either compound alone caused partial but incomplete inactivation within the same time periods. A population of 10(5) CFU/cm2 of 1-day adherent cells on stainless steel was completely inactivated within 25 min, but with the highest concentrations of the combined chemicals, i.e., 1% acetic acid and 100 microg/ml of monolaurin. The combined chemical treatment again synergistically produced greater inhibition. A 10(6)-CFU/cm2 population of 7-day adherent cells was not completely inactivated within 25 min of exposure, although counts did decline. The results demonstrate increased resistance of attached L. monocytogenes to acetic acid and monolaurin and show that resistance increased with culture age. Combinations of organic acids and monolaurin might be considered as sanitizers of food contact surfaces, but activities of such combinations are likely to be less than other commonly used sanitizers.

The effect of pH on the minimum inhibitory concentration (MIC) of glycerol monolaurate (monolaurin) against four strains of Listeria monocytogenes at 35°C in tryptic soy broth supplemented with 0.6% yeast extract was investigated. Our results demonstrate that the MIC of monolaurin was lower than MIC values reported for other common food antimicrobials such as potassium sorbate, tertiary butylhydroquinone, propyl paraben, and butylated hydroxyanisole. The MIC of monolaurin was reduced by decreasing the pH value of the medium. A 3-fold MIC reduction occurred when the pH decreased from pH 7.0 (10 μg/ml) to pH 5.0 (3 μg/ml).