• Journal of Environmental Health Sciences
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• v.38 no.1
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• pp.73-81
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• 2012

Objectives: With the goal of quantifying the risk of children contracting gastroenteritis while playing at interactive waterscape facilities and evaluating the adequacy of current water quality regulations, risk assessment was performed with Escherichia coli as pathogen. Methods: Abundances of E. coli in the waters of interactive water features in South Korea were acquired from survey reports. A gamma distribution describing the volume of water swallowed by children during swimming activities was adopted. Exposure rate and risk were calculated by Monte Carlo simulation and dose-response models for various pathogenic E. coli. Results: E. coli was detected in 25 out of 40 facilities, with range of ~1,600 CFU/100 ml. The abundance fitted an exponential distribution. Simulated exposures ranged ${\sim}1.9{\times}10^{10}$ CFU, varying greater along E. coli abundance than the volume of water. Risk of children being infected by enterohemorrhagic E. coli was high, with range of ~0.85. When E. coli abundance was <200 CFU/100 ml, which is the current government threshold, the risk decreased to <0.43. Although the guideline successfully reduced the risk of adults being infected by a less virulent E. coli strains (<0.03), the risk for children could not be quantified due to lack of dose-response models for those pathogens for children. Conclusions: Under the current guideline, children are at risk of being infected if water is contaminated with by enterohemorrhagic E. coli. For other E. coli strains, the risk appears to be considerably less. The result warrants need for developing dose-response models for children for each pathogenic E. coli strain.

• Biomedical Science Letters
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• v.4 no.1
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• pp.43-56
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• 1998

A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-I.II), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) O157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli O157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-I.II), and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 was found to be 2.5$\times$10$^{6}$ of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC O157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC O157:H7.

• Food Science of Animal Resources
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• v.37 no.4
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• pp.579-592
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• 2017

This study assessed the quantitative microbial risk of non-enterohemorrhagic Escherichia coli (EHEC). For hazard identification, hazards of non-EHEC E. coli in natural and processed cheeses were identified by research papers. Regarding exposure assessment, non-EHEC E. coli cell counts in cheese were enumerated, and the developed predictive models were used to describe the fates of non-EHEC E. coli strains in cheese during distribution and storage. In addition, data on the amounts and frequency of cheese consumption were collected from the research report of the Ministry of Food and Drug Safety. For hazard characterization, a doseresponse model for non-EHEC E. coli was used. Using the collected data, simulation models were constructed, using software @RISK to calculate the risk of illness per person per day. Non-EHEC E. coli cells in natural- (n=90) and processed-cheese samples (n=308) from factories and markets were not detected. Thus, we estimated the initial levels of contamination by Uniform distribution ${\times}$ Beta distribution, and the levels were -2.35 and -2.73 Log CFU/g for natural and processed cheese, respectively. The proposed predictive models described properly the fates of non-EHEC E. coli during distribution and storage of cheese. For hazard characterization, we used the Beta-Poisson model (${\alpha}=2.21{\times}10^{-1}$, $N_{50}=6.85{\times}10^7$). The results of risk characterization for non-EHEC E. coli in natural and processed cheese were $1.36{\times}10^{-7}$ and $2.12{\times}10^{-10}$ (the mean probability of illness per person per day), respectively. These results indicate that the risk of non-EHEC E. coli foodborne illness can be considered low in present conditions.

• Journal of Preventive Medicine and Public Health
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• v.36 no.1
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• pp.77-84
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• 2003

Objectives : Two related cases of Hemolytic-Uremic Syndrome (HUS) were reported to the Korea National Institute of Health in May, 2001. Shiga toxin 2 genes were detected in both stool samples. We suspected an enterohemorrhagic Escherichia coli (EHEC) infection as the cause of the HUS, and conducted an investigation to find the source of the infection and its route of transmission. Methods : We peformed case investigations on these two related HUS cases, and obtained interviews and rectal swabs form the family members and other close contacts. Additionally, we peformed rectal swabs on the cattle raised by the household of the index patient. Results : We found a 20 month old index patient and a 6 year-old cousin had developed HUS, where there had been a 2 day history of contact with the index, and bacteriological examinations for these two patients revealed, indistinguishably, the same E. coli O171. The grandmother of the index patient was found to be asymptomatic, but E. coli O26 was isolated. We also found a probable case in the mother of the cousin. She reported a history of contact with the index, and developed bloody diarrhea of 3 days duration. The test results for the cattle revealed E. coli O26 in one cow, and E. coli O26 and O55 in another. E. coli O26, which was isolated in both cows and the grandmother of the index, were indistinguishably the same. Conclusions : We found that the E. coli O26 in the grandmother had originated from the cows, and that the E. coli O171 found in the index patient had been transmitted to the cousin through person-to-person contact.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2003.10a
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• pp.30-30
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• 2003

Escherichia coli strains associated with diarrhea have been divided into the following six major categories on the basis of pathogenic mechanisms: enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAggEC) and diffusely adherent E. coli (DAEC).$\^$15,18/ EPEC, EAggEC, and DAEC strains were classified by their ability to produce distinct patterns of adherence to cultured epithelial cells in virto: localized (LA), aggregative (AA), and diffuse (DA) adherence. The objective of this study was to investigate the relationship of the adherence patterns with AIDA-positive E. coli isolated from diarrheic pigs. (omitted)

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.130-136
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• 2021

Persister cell formation and biofilms of pathogens are extensively involved in the development of chronic infectious diseases. Eradicating persister cells is challenging, owing to their tolerance to conventional antibiotics, which cannot kill cells in a metabolically dormant state. A high frequency of persisters in biofilms makes inactivating biofilm cells more difficult, because the biofilm matrix inhibits antibiotic penetration. Fatty acids may be promising candidates as antipersister or antibiofilm agents, because some fatty acids exhibit antimicrobial effects. We previously reported that fatty acid ethyl esters effectively inhibit Escherichia coli persister formation by regulating an antitoxin. In this study, we screened a fatty acid library consisting of 65 different fatty acid molecules for altered persister formation. We found that undecanoic acid, lauric acid, and N-tridecanoic acid inhibited E. coli BW25113 persister cell formation by 25-, 58-, and 44-fold, respectively. Similarly, these fatty acids repressed persisters of enterohemorrhagic E. coli EDL933. These fatty acids were all medium-chain saturated forms. Furthermore, the fatty acids repressed Enterohemorrhagic E. coli (EHEC) biofilm formation (for example, by 8-fold for lauric acid) without having antimicrobial activity. This study demonstrates that medium-chain saturated fatty acids can serve as antipersister and antibiofilm agents that may be applied to treat bacterial infections.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.5-14
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• 2010

Enterohemorrhagic Escherichia coli O157:H7 is a major foodborne pathogen causing severe disease in humans worldwide. Healthy cattle are a reservoir of E. coli O157:H7, and bovine food products and fresh produce contaminated with bovine waste are the most common sources for disease outbreaks in the United States. E. coli O157:H7 also survives well in the environment. The abilities to cause human disease, colonize the bovine gastrointestinal tract, and survive in the environment require that E. coli O157:H7 adapt to a wide variety of conditions. Three major virulence factors of E. coli O157:H7 have been identified including Shiga toxins, products of the pathogenicity island called the locus of enterocyte effacement, and products of the F-like plasmid pO157. Among these virulence factors, the role of pO157 is least understood. This review provides a board overview of E. coli O157:H7 with an emphasis on pO157.

• Microbiology and Biotechnology Letters
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• v.48 no.2
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• pp.121-128
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• 2020

Diarrhea is a major public health concern associated with pathogenic Escherichia coli infections. Food-borne pathogenic E. coli can lead to large diarrheal outbreaks and hence, there is a need to estimate the frequency of pathogenic E. coli load in the various types of meat available in markets. In the present study, we classified and characterized diarrheagenic E. coli isolates collected from 399 raw meat samples from retail sources in Korea. Shiga toxin-producing E. coli (STEC) were detected in 11 (9.7%) samples, including nine strains (8.0%) in beef and two strains (1.8%) in chicken. The frequency of the detected virulence markers were as follows: astA, 28.3%; escV,18.6%; eaeA,17.7%; ent, 7.0%; EHEC-hly, 4.4%; stx1, 3.5%; and stx2, 3.5%. We did not observe any typical EPEC, EIEC, or ETEC virulence determinants in any of the samples. The STEC serotype O26 was detected in one sample, but no other serogroups (O91, O103, O128, O157, O145, O111, and O121) were found. Further research is needed to better understand the virulence mechanism of STEC serotypes, their ecology, and prevalence in animals, food, and the environment. These results will help improve risk assessment and predict the sources of food poisoning outbreaks.

• 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.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.401-407
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• 2014

Quorum sensing and the stringent response are well-known regulation systems for the expression of virulence genes in enterohemorrhagic Escherichia coli (EHEC). However, how these two systems interact is not well known. E. coli strains with mutations in two regulation systems, ${\Delta}luxS$ (ECM101) and ${\Delta}luxS{\Delta}relA{\Delta}spoT$ (ECM201), and the ${\Delta}luxS$ complement strain to ECM201 (ECM202) were created from EHEC O157:H7 EDL933 to investigate how the regulatory systems interact. The phenotypic changes of the mutant strains were characterized and compared with the wild type. The mutant strains exhibited no obvious growth defects, although acid resistance and cellular cytotoxicity were decreased significantly in all the mutant strains. Phenotypic characterization revealed that mutations in the stringent response system (ECM201 and ECM202) influenced the metabolic (defective utilization of arabinose and L-sorbose) and enzymatic activities (decreased trypsin activity, and increased ${\alpha}$-glucosidase activity). In contrast, the quorum sensing system mutant (ECM101) did not display these phenotypes. The motility of the quorum sensing system mutant (ECM101) was unchanged, but mutation in the stringent response system influenced the motility. Our results suggest that quorum sensing interacts with the stringent response regulation system.