• Journal of Animal Science and Technology
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• v.62 no.4
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• pp.543-552
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• 2020

For efficient prevention and treatment of enteric colibacillosis, understanding about latest virulence factors and antimicrobial resistance of Escherichia coli is essentially needed. The aim of this study was to survey antimicrobial resistance and determine the prevalence of fimbriae and enterotoxin genes among 118 pathogenic E. coli isolates obtained from Korean pigs with diarrhea between 2016 and 2017. The genes for the toxins and adhesins were amplified by polymerase chain reaction (PCR). The susceptibility of the E. coli isolates to antimicrobials were tested using the standard Kirby-Bauer disk diffusion method. The most prevalent fimbrial antigen was F18 (40.7%), followed by F4 (16.9%), and the most prevalent combinations of toxin genes were Stx2e (21.2%), STb:EAST-1 (19.5%), and STa:STb (16.9%), respectively. Among the pathotypes, enterotoxigenic E. coli (ETEC) was the most predominant (67.8%), followed by Shiga-toxin producing E. coli (STEC, 23.7%). We confirmed high resistance rates to chloramphenicol (88.1%), tetracycline (86.4%), streptomycin (86.4%), and ampicillin (86.4%). And the majorities of isolates (90.7%) showed multi-drug resistance which means having resistance to 3 or more subclasses of antimicrobials. Results of this study can be a source of valuable data for investigating the epidemiology of and control measures for enteric colibacillosis in Korean piggeries.

• 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 Animal Science and Technology
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• v.61 no.2
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• pp.55-60
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• 2019

Colibacillosis is one of the major health problems in young piglets resulting in poor health and death caused by Escherichia coli producing F18 pili and Shiga toxin 2e. It is pivotal to reduce colibacillosis in weaned piglets to enhance production performance. In this study, we evaluated synbiotics as the gut health improvement agents in the mouse model challenged with Shiga toxin-producing E. coli (STEC) isolated from piglets. Prebiotic lactulose was formulated with each $5.0{\times}10^6CFU/mL$ of Pediococcus acidilactici GB-U15, Lactobacillus plantarum GB-U17, and Lactobacillus plantarum GB 1-3 to produce 3 combinations of synbiotics. A total of 40 three weeks old BALB/c mice were randomly assigned to 4 groups (n = 10): a control group and 3 synbiotics treated groups. Each treatment groups were daily administrated with $5.0{\times}10^6CFU/mL$ of one synbiotics for the first week, and every 3 days during the second week. All the mice were challenged with $8.0{\times}10^8CFU/mL$ of STEC 5 days after animals began to receive synbiotics. Mice treated with synbiotics based on Pediococcus acidilactici GB-U15 and Lactobacillus plantarum GB-U17 significantly improved daily weight gain compared to mice in other groups. While mice treated with GB-U15 showed better fecal index, no significant differences were observed among groups. Gross lesion and histopathological evaluations showed that mice treated with GB-U15 moderately improved recovery from STEC infection. In conclusion, our results suggest that the synbiotics formulated with lactulose and Pediococcus acidilactici GB-U15 have potential benefits to prevent and improve colibacillosis in weaned piglets.

• Food Science of Animal Resources
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• v.26 no.2
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• pp.263-268
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• 2006

Probiotics including Lactobacillus acidophilus, refer to a group of nonpathogenic organisms that protect the human host against gastrointestinal(GI) infections by pathogenic bacteria such as Shiga toxin-producing E. coli(STEC). In the study, the inhibitory effects of STEC ATCC 43894 adhesion by L. acidophilus A4 was investigated on the HT-29 epithelial cells. Specific proteins regulated by cell Iysates of L. acidophilus A4 on STEC ATCC 43894 were also characterized by proteomic analysis. Both cell mass and Iysate of L. acidophilus A4 have exhibited the profound inhibitory activity on the HT-29 cells(about 1.5 log scale reduction). Two-dimensional gel electrophoresis(2-DE) revealed seven proteins that were up-regulated by cell Iysates of L. acidophilus A4 and three proteins that were down-regulated. In addition, three protein spots were only detected in the presence of cell Iysates. These results suggest that inhibitory effects of STEC adhesion by L. acidophilus may be due to the regulation of specific protein of STEC.

• Clinical and Experimental Pediatrics
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• v.46 no.2
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• pp.143-153
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• 2003

Purpose : Infection with Shiga-like toxin (SLT)-producing Escherichia coli, an emerging human pathogen found particularly in young children under 5 years of age, causes a spectrum of illnesses with high morbidity and mortality, ranging from diarrhea to hemorrhagic colitis and hemolytic uremic syndrome. Host mediators play an important role in the pathogenesis of SLT-I toxicity. The experiments described here were designed to investigate the effect of SLT-I on TNF-${\alpha}$ production and to understand the effect of TNF-${\alpha}$ on GB3 expression. We also further examine the relationship between the Gb3 level and the differential susceptibility of cells to the cytotoxic action of SLT-I. Methods : The effect of purified SLT-1 from E. coli O157 : H7 (ATCC 43890) on tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) production in Raw264.7 cells was investigated. Many mediators regulate endothelial cell membrane expression of the glycolipid globotriaosyleramide (Gb3), which serves as the toxin receptor, suggesting that the host response to the toxin or other bacterial products may contribute to pathogenesis by regulating target cell sensitivity to the toxins. Therefore, the relationships between Gb3 expression and cytotoxicity against SLT-I on three types of cells were evaluated. Results : Detectable levels of TNF-${\alpha}$ were produced as early as six hours after induction and continued to increase during 48 hours by SLT-I. It was also found that Vero cells and dendritic cells (DC2.4 cells) expressed high levels of Gb3, 83% and 68%, respectively, and that Raw264.7 cells had a low level of Gb3 (29%) and appeared refractory to cytotoxicity against SLT-I. Vero cells and DC2.4 cells expressing high levels of Gb3 were highly susceptible to SLT-I. Furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. Conclusion : These results strongly suggest that the expression of Gb3 is necessary but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its Gb3 receptors in the pathogenesis of E. coli O157 infection.

• 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.

• Journal of Microbiology and Biotechnology
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• v.30 no.10
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• pp.1552-1558
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• 2020

With an increase in the consumption of non-heated fresh food, foodborne shiga toxin-producing Escherichia coli (STEC) has emerged as one of the most problematic pathogens worldwide. Endolysin, a bacteriophage-derived lysis protein, is able to lyse the target bacteria without any special resistance, and thus has been garnering interest as a powerful antimicrobial agent. In this study, rV5-like phage endolysin targeting E. coli O157:H7, named as LysECP26, was identified and purified. This endolysin had a lysozyme-like catalytic domain, but differed markedly from the sequence of lambda phage endolysin. LysECP26 exhibited strong activity with a broad lytic spectrum against various gram-negative strains (29/29) and was relatively stable at a broad temperature range (4℃-55℃). The optimum temperature and pH ranges of LysECP26 were identified at 37℃-42℃ and pH 7-8, respectively. NaCl supplementation did not affect the lytic activity. Although LysECP26 was limited in that it could not pass the outer membrane, E. coli O157: H7 could be effectively controlled by adding ethylenediaminetetraacetic acid (EDTA) and citric acid (1.44 and 1.14 log CFU/ml) within 30 min. Therefore, LysECP26 may serve as an effective biocontrol agent for gram-negative pathogens, including E. coli O157:H7.

• Korean Journal of Veterinary Service
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• v.46 no.4
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• pp.325-334
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• 2023

Edema disease (ED) in pigs is enterotoxemia caused by Shiga toxin type 2e (Stx2e)-producing Escherichia coli (STEC) and frequently occurs in young piglets. Therefore, ED causes enormous economic losses in pig farms. In this study, a modified Stx2e (mStx2e) antigen was expressed and purified using commercial E. coli expression system. Monoclonal antibody was serviced by Ynto Ab Inc., using Phage Display Technique. Anti-Stx2e antibodies in piglets were measured by indirect ELISA using mStx2e antigens. Naive Stx2e in piglets were detected by Sandwich ELISA using Stx2e-monoclonal antibodies and commercial Stx2e-polyclonal antibodies. Among 3,480 piglets, anti-Stx2e antibodies were observed in 2,573 piglets. The 49.4% among 830 piglet serum samples possessed 0.625 ㎍/mL or more of Stx2e proteins. The 18.3% of 830 sera had 0.313 ㎍/mL of Stx2e proteins. The 32.3% of 830 samples held 0.156 ㎍/mL or less of Stx2e proteins. These results show that indirect ELISA using mStx2e antigen and Sandwich ELISA using Stx2e-monoclonal and polyclonal antibodies can be useful to detect ED in piglets.

• Korean Journal of Veterinary Service
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• v.44 no.4
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• pp.271-281
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• 2021

Pathogenic Escherichia coli (E. coli) is one among the most important agents of diarrhea in calves. From January to December 2021, 108 isolates from feces of calves with diarrhea were investigated for enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), shiga toxin-producing E. coli (STEC), enteroaggregative E. coli (EAEC), and enteroinvasive E. coli (EIEC) using real-time PCR. In addition, the genes for F5, F17 and F41 fimbriae were detected by PCR. The most frequently isolated pathotypes were EPEC/STEC (29 isolates), and ETEC/EPEC/STEC (29 isolates). ETEC/EPEC, and ETEC/STEC were also found in 10 isolates. EPEC, STEC, and ETEC were detected in 13, 11, and 6 respectively. EAEC, and EIEC was not detected. Antimicrobial resistance test was carried out by agar disc diffusion method with 14 antimicrobials. Among 108 pathogenic E. coli isolates, 107 isolates were resistant to at least one of 14 antibiotics used in this study, 99 (91.7%) were resistant to two or more antimicrobials, and a single remarkable isolate was resistant to 14 antimicrobials. The isolates were primarily resistant to penicillins, streptomycin, tetracycline, ceftiofur, Trimethoprim/sulfamethoxazole, Kanamycin, and Ciprofloxacin. The high rate of resistance in pathogenic E. coli, sometimes to multiple drugs, may complicate future options for treating human infections. These results may bu used for diagnosis and therpeitic purposes in calves with diarrhea.

• Childhood Kidney Diseases
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• v.6 no.1
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• pp.102-108
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• 2002

We report two cases of hemolytic uremic syndrome (HUS) associated with Escherichia coli O114. Two cases were similar and showed the same clinical courses. After prodrome of diarrhea and vomiting lasting 1-2 days, azotemia persisted for about 10 days, and during that period, the patients were on peritoneal dialysis. They recovered without any sequelae after about 15 days. Direct multiplex PCR of stool culture revealed eae and stx2 gene and the result of ELISA done on the colony positive of one gene confirmed Escherichia coli O114. This is the first report of HUS associated with Escherichia coli O114. We recommend, Shiga toxin producing bacterial Infection must be considered and efforts should be made to scrutinize the organism in all diarrhea-prodrome HUS patients.(J Korean Soc Pediatr Nephrol 2002 ;6 : 102-8)