• Biomedical Science Letters
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• v.8 no.3
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• pp.107-113
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• 2002

Enterohemorrhagic Escherichia coli (EHEC) strains of serotype O157:H7 have been shown to colonize the intestinal epithelial cell by the attaching and effacing (AE) mechanism. The AE lesion is mediated by an intimin, of which production and expression are controlled by a 3-Kb eaeA gene located EHEC chromosomal DNA. If the eaeA gene is mutated, EHEC O157:H7 strains lose capacity of adhesion to intestinal epithelial cells. In this study, a 891 bp of the 3'-end region of a gamma intimin was amplified by polymerase chain reaction (PCR). The PCR product was inserted into pSTBlue-1 cloning vector and transformed into DE3 (BL21) competent cell. After plasmid mini-preparation and restriction enzyme digestion of eaeA/891-pSTBlue-1 vector, target eaeA gene was re-inserted into pET-28a expression vector and was transformed. Then the expression of recombinant eaeA/891 (891 bp) gene was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG). The expression of the 40-KDa recombinant protein was identified in SDS-PAGE and confirmed by immunoblotting using the His.Tag$^{\circledR}$ and T$_{7}$.Tag$^{\circledR}$ monoclonal antibody. This recombinant protein expressed by eaeA gene could be applied in further studies on the mechanisms of E. coli O157:H7 infection and the development of recombinant vaccine.

• Preventive Nutrition and Food Science
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• v.6 no.1
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• pp.6-9
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• 2001

Minimal inhibitory concentrations (MIC) and antimicrobial effects of glycerol monolaurate (monolaurin) and organic acids, either alone or in combination, against Escherichia coli O157:H7 in tryptic soy broth were determined. MIC values of monolaurin(ML), acetic (AA), citric (CA), lactic (LA) and hydrochloric acid (HCl) were 300$\mu\textrm{g}$/mL (0.03%), 1250$\mu\textrm{g}$/mL(0.125%), 5000$\mu\textrm{g}$/mL(0.5%), 2500$\mu\textrm{g}$/mL(0.25%) and 2500$\mu\textrm{g}$/mL(0.25), respectively. When 150$\mu\textrm{g}$/mL of ML was combined with 50$\mu\textrm{g}$/mL AA, 250$\mu\textrm{g}$/mL HCl and 125$\mu\textrm{g}$/mL LA, the combined agents did not increase the inhibitory effect of the most active single compound alone. This result indicates that there was little interaction between ML and A, HCl and LA. However, the combination of 150$\mu\textrm{g}$/mL ML and 250$\mu\textrm{g}$/mL CA synergistically inhibited growth of E. coli O157:H7. The present study showed that the nature of combined antimicrobial response against E. coli O157:H7 was complex, but this information would be useful for determining interaction that could compromise effectiveness in food systems.

• Journal of the East Asian Society of Dietary Life
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• v.22 no.1
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• pp.109-119
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• 2012

This study analyzed the antibacterial activity of the ethanol extract of Schizandra chinensis Baillon against food pathogenic microorganisms to determine its capabilities as a natural antimicrobial agent. A paper disc diffusion test, minimum inhibitory concentration (MIC) determination, and time-kill assay showed that the ethanol extract strongly inhibits the growth of Listeria monocytogenes, Bacillus cereus, Escherichia coli O157:H7, and Pseudomonas aeruginosa. Release of cytoplasmic ${\beta}$-galactosidase was detected in E. coli, E. coli O157:H7, S. aureus, and P. aeruginosa treated with the ethanol extract. An increase of outer membrane permeability caused by the ethanol extract was also observed. An outward flow of cell constituents was detected in the Gram negative strains treated with the ethanol extract. These results imply that the inner and outer membranes of cells were partially destroyed and cell constituents were released by the treatment of the S. chinensis Baillon ethanol extract. The results of this study indicate that ethanol extract of S. chinensis Baillon evidences a fairly good antibacterial effect.

• Korean journal of food and cookery science
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• v.24 no.4
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• pp.445-451
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• 2008

This study compared the effects of chlorine dioxide and a commercial chlorine sanitizer for inhibiting foodborne pathogens, including Salmonella enterica serovar Typhimurium, Listeria monocytogenes, and Escherichia coli O157 : H7, on lettuce leaves. The lettuce samples were inoculated with each cocktail of the three strains, and were then treated with chemical sanitizers [distilled water, 100 ppm commercial chlorine and 50 ppm, 100 ppm, 200 ppm chlorine dioxide ($ClO_2$)] for 1 min, 5 min, and 10 min at room temperature($22{\pm}2^{\circ}C$). Following inoculation of the leaves, initial populations of E. coli O157:H7, L. monocytogenes, and S. Typhimurium were approximately 5.54, 4.47, and 5.12 log CFU/g, respectively these levels were not significantly reduced by the treatment with water,whereas the 100 ppm commercial chlorine sanitizer treatment and $ClO_2$ (at all tested concentrations) were effective at reducing levels of all three pathogens. The treatment of 200 ppm $ClO_2$ for 10 min was most effective at inhibiting the three pathogens, and reduction levels of E. coli O157 : H7, L. monocytogenes, and S. Typhimurium were 2.28, 1.95, 1.76 log, respectively. The inhibitory effect of $ClO_2$ increased with increasing treatment concentration of $ClO_2$, but there was no significant difference by the treatment times. When chemically injured cells of E. coli O157 : H7 and L. monocytogenes and S. Typhimurium were examined by SPRAB and selective overlay methods, respectively, it was observed that the commercial chlorine sanitizer generated greater numbers of injured L. monocytogenes than the $ClO_2$ treatment. From the overall results, $ClO_2$ was more effective at inhibiting pathogenic bacteria compared to the commercial chlorine sanitizer therefore, it has potential to be utilized as an alternative sanitizer to increase the microbial safety of fresh produce.

• Journal of Food Hygiene and Safety
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• v.30 no.1
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• pp.92-97
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• 2015

The aim of this study is to examine the removal efficiency of pathogen (Escherichia coli O157:H7 and Salmonella typhimurium) on meat and fish products (packing condition: vacuum or not and storage temperature: $4^{\circ}C$ or $-20^{\circ}C$) repeatedly exposed at low-dose gamma irradiation. In case of meat products (beef and chicken), E. coli O157:H7 was not observed at the level of 2 kGy single gamma irradiation and 0.5 kGy repeated gamma irradiation and S. Typhimurium was not observed at the level of 2 kGy single gamma irradiation and 1 kGy repeated gamma irradiation. In case of fish products, E. coli O157:H7 and S. Typhimurium were not detected at the level of 0.5 kGy single and repeated gamma irradiation. These results showed that microorganisms on fish products were more efficiently removed than those of meat products with low-dose gamma irradiation. Generally, each packing condition made no difference. However, the products (fish and meat) stored at $-20^{\circ}C$ needed more higher dose gamma irradiation than products at $4^{\circ}C$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.4
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• pp.577-584
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• 2016

The objectives of this study were I) to compare the acid resistance (AR) of seven non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups, including O26, O45, O103, O111, O121, O145, and O157:H7 STEC isolated from various sources, in 400 mM acetic acid solution (AAS) at pH 3.2 and $30^{\circ}C$ for 25 min with or without glutamic acid and II) to determine strain survival upon exposure to simulated gastric fluid (SGF, pH 1.5) at $37^{\circ}C$ for 2 h after acid adaptation in apple, pineapple, orange, and strawberry juices at pH 3.8, $4^{\circ}C$ and $20^{\circ}C$. Results show that the O111 serogroup strains had the strongest AR (0.12 log reduction CFU/mL) which was very similar to that of O157:H7 STEC (P>0.05), compared to other serogroups in AAS without glutamic acid, whereas O26 serogroup strains showed the most sensitive AR. However, there was no significant (P>0.05) difference of AR among seven serogroups in AAS with glutamic acid. In the SGF study, 05-6545 (O45:H2), 08023 (O121:H19), and 03-4669 (O145:NM) strains adapted in fruit juices at $4^{\circ}C$ and $20^{\circ}C$ displayed enhanced survival with exposure to SGF for 60 min compared to 06E0218 (O157:H7) strains (P<0.05). In addition, 4 STEC strains adapted in pineapple juice at $4^{\circ}C$ showed enhanced survival with exposure to SGF for 60 min compared to those strains acid-adapted in the other fruit juices. Generally, adaptation at $4^{\circ}C$ in fruit juices resulted in significantly enhanced survival levels compared to acid-adapted at $20^{\circ}C$ and non-adapted conditions. The AR caused by adaptation in fruit juices at low temperature may thus increase survival of non-O157 STEC strain in acidic environments such as the gastrointestinal tract. These results suggest that more careful strategies should be provided to protect against risk of foodborne illness by non-O157 STEC.

• Journal of Food Hygiene and Safety
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• v.12 no.3
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• pp.188-194
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• 1997

The growth inhibition effect of Orally administrated yogurt ACE and Metchnikoffupon E. coli O157:H7 and S. typhimurium inoculated into gastric lumen of rabbits was in vestigated. The rabbits challenged with each 1 $m\ell$ of suspension containing 108 CFU/$m\ell$ of the pathogens were divided into 4 groups by the interval of yogurt administration: A group; preadministrated 7 days before inoculation of the pathogens and fed daily; B group; administrated daily after inocjlation of the pathogens, C group; administrated every 3 days after inoculation of the pathogens; Control group, not fed after inoculation of the pathogens. Each 3 $m\ell$ of yogurt containing 109 CFU/$m\ell$ was orally administrated into rabbits. All yogurt administrated groups (A, B, c) chowed growth ingibition effect on E. coli O157:H7 in one day after inoculation of the pathogen by the level of 0.8~1.0 log CFU/g, compared with the result differences between the control group and the yogurt administrated groups. In the control group after 5 days of inoculation, the number of colonized pathogens was 105~106 CFU/g, whereas 103~104 CFU/g was detected in the yogurt administrated groups. After 10 days of inoculation, the viable pathogen number per gram (g) of the rabbit feces was 103 CFU/g in the control group, whereas the number below 101 CFU/g was detected in the group A, and 102 CFU/g in the control group, B and C. The growth inhibition effect of yogurt administration on E. coli O157:H7 was highly increased in the order of A, B, and C group. The same effect on S. typhimurium was observed at the level of 2 log CFU/g in the Metchnikoff yogurt administrated groups, compared with the control group result in one day after inoculation of the pathogen. In 7 days after inoculation of the pathogen, the viable number was increasingly decreased, and finally after 15 days no viable cell of S. typhimurium was discharged into the fecal samples in the group A, and the mean level of 10* CFU/g was detected in the group B, but there was no growth inhibition effect in the group C. The growth inhibition effect on S. typhimurium was observed at the same level of viable cell number between the yogurt ACE administrated groups and the control group in 5 days after inoculation. But, after 10 days of inoclation the viable cell number was started to decrease, and the viable cell of S. typhimurium was not discharged from rabbit intestinal contents after 15 days of inoculation in the yogurt ACE administrated groups. In such a case that yogurt was administrated in order to prevent the pathogens, pre-administration on a daily basis one week before inoculation of the pathogens exerted considerable effect in growth inhibition. In comparison with two kinds of yogurt tested in this study, the growth inhibition effect on two kinds of pathogens was observed more highly in the Metchnikoff administated group than the ACE administrated group.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.679-682
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• 2008

A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

• Korean Journal of Food Science and Technology
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• v.53 no.6
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• pp.819-822
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• 2021

To enhance phage endolysin-mediated cell wall disruption of Escherichia coli O157:H7, the cells were co-treated with aromatic compounds, namely thymol or eugenol, found in essential oils and endolysin LysECP26. Interestingly, the minimal inhibitory concentrations of LysECP26 was four times lower when used in combination with either of the two compounds than when it was used alone. This synergistic activity was also confirmed by viable cell counting. Within 1 h of LysECP26 and eugenol or thymol co-treatment to the cells, there was a 2.3 or 3.8 log CFU/mL reductions, respectively. Additionally, field emission scanning electron microscopy showed cell wall disruption and severe morphological alterations of the cells in case of the combination treatments. Therefore, endolysin and thymol or eugenol co-treatment can help in developing efficient bio-control strategies against gram-negative pathogen E. coli O157:H7.

• Korean Journal of Environmental Agriculture
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• v.38 no.4
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• pp.254-261
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• 2019

BACKGROUND: Agricultural water is known to be one of the major routes in bacterial contamination of fresh vegetable. However, there is a lack of fundamental data on the microbial safety of agricultural water in Korea. METHODS AND RESULTS: We investigated the density of indicator bacteria in the surface water samples from 31 sites collected in April, July, and October 2018, while the groundwater samples were collected from 20 sites within Jeollabuk-do in April and July 2018. In surface water, the mean density of coliform, fecal coliform, and Escherichia coli was 2.7±0.55, 1.9±0.71, and 1.4±0.58 log CFU/100 mL, respectively, showing the highest bacterial density in July. For groundwater, the mean density of coliform, fecal coliform, and E. coli was 1.9±0.58, 1.4±0.37, and 1.0±0.33 log CFU/ 100mL, respectively, showing no significant difference between sampling time. The survival of E. coli O157:H7 were prolonged in water with higher organic matter contents such as total nitrogen (TN), and nitrate-nitrogen (NO₃-N). The reduction rates of E. coli O157:H7 in the water showed greater in order of 25, 35, 5, and 15℃. CONCLUSION: These results can be utilized as fundamental data for prediction the microbiological contamination of agricultural water and the development of microbial prevention technology.