• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.1
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• pp.76-83
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• 1992

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin (ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. ST producing E. coli KM-7 strain was isolated from the swine and molecular cloning of ST gene of KM-7 strain. Transformant eKT-53 $(ST^+,\;LT^-)$ was selected by infant mouse assay (IMA). The culture supernatant of eKT-53 strain was performed purification by multipled steps. The culture supernatant (crude ST) was purified by sequentially applying batch adsorption chromatography on Amberlite XAD-2 resin, ion exchange chromatography on DEAE-Sephacel anion exchanger, gel filtration chromatography on Bio-Gel P-6 and preparative polyacrylamide slab gel electrophoresis. About 113-fold purification was achieved with a yield of about 11% of crude ST and the minimum effective dose(MED) of this purified ST was about 2.8ng in IMA. Homogeneity of purified ST was demonstrated by showing a single band in analytical SDS polyacrylamide disc gel electrophoresis.

• Korean Journal of Veterinary Research
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• v.27 no.2
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• pp.259-267
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• 1987

Enterotoxigenic E. coli (ETEC) cause an acute diarrhea (white scour) in both animals and humans. The disease process initially involves the adherence and colonization of the mucosal surface of the small intestine, followed by the elaboration of a heat-labile enterotoxin (LT) and/or heat-stable enterotoxin (ST). Intestinal adherence or colonization by ETEC is generally mediated by a specific surface-associated pilus (fimbrial) antigen that endows the bacteria with the capacity to adhere to epitherial cell surface. Fourteen monoclonal antibodies (MAbs) directed against pili antigens of ETEC were obtained by cell fusion/hybridoma technique. They were characterized by indirect immunofluorescence assay (IFA), and divided into four groups: specific to K99 antigen (group 1), cross-reactive with K99 and F41 antigens (group 2), specific to K88 antigen (group 3) and specific to 987P and K88 antigens (group 4), respectively. These MAbs demonstrated the distinct pili (K) antigens on the surface of ETEC by IFA, and could be utilized as diagnostic reagent for the identification of ETEC. When eighty-seven field isolates of E. coli from piglet with diarrhea were tested by group 3 MAb, fourty-two strains (48.3%) has K88 pilus antigen suggesting that this is one of the major pilus antigen of ETEC present in fifeld.

• Journal of Nutrition and Health
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• v.34 no.2
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• pp.150-157
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• 2001

Intestinal absorption of dietary taurine is one of the regulatory component maintaining taurine homeostasis along with renal reabsorption, bile acid conjugation and secretion, and de nobo synthesis of taurine in mammals. Recent observations of decreased enterocytic levels of taurine in response to trauma, infection and surgical insults, postulate the possibility that intestinal taurine absorption might be impaired in such stressed conditions. The aim of the present study was to evaluate changes in enterocytic taurine transporter activity using the human intestinal colon carcinoma cell line, HT-29, in various stress-induced conditions. Pretreatment of the HT-29 cells with dexamethasone, a stress hormone(0.1,1,10 or 100$\mu$M) for 3 hrs, or with E coli heat-stable enterotoxin(10, 100, or 200nM) for 30 minutes in order to induce the condition of enterotoxigenic infection did not influence taurine uptake as compared to the value found in control cells. In contrast, pretreatment of the cells with cholera toxin(10, 100, 500, or 1000ng/ml)for 3hr or 24hr significantly decreased taurine uptake by HT-29 cells to 40~50% of the value found in untreated control cells. Kinetic studies of the taurine transporter activity were conducted in control and cholera toxin treated HT-29 cells with varying taurine concentrations(2~60$\mu$M) in the uptake medium. Pretreatment of the cells with cholera toxin(100ng/ml) for 3hr did not influence the Vmax, but resulted in a 55% increase in the Michaelis-Menten constant(Km) of the taurine transporter compared to those in control cells. These results suggest that cholera toxin-induced reduction in taurine transporter activity in HT-29 cells is associated with decreased affinity of the taurine transporter without altering the amount of transporter protein. Intestinal taurine absorption appears to be reduced in the condition of water-borne diseases caused by bacteria such as V. cholerae. This might influence the taurine status of infants and young children more readily, an age group in which the prevalence of intestinal infection is high and the role of intestinal absorption is crucial for maintaining the body taurine pool. (Korean J Nutrition 34(2) : 150-157, 2001)

• The Journal of the Korean Society for Microbiology
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• v.19 no.1
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• pp.65-71
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• 1984

The author attempted serological typing with the slide agglutination and the production of heat-labile enterotoxin (LT) and heat-stable enterotoxin (ST) by the enterotoxingenic eschenchia coli 446 strains isolated from diarrheal patients. The enterotoxingenic E.coli, producing LT and/or St, were detected by use of assays in the reversed passive hemagglutination(RPHA) and the suckling mouse method. The results obstained were as follows: 1. Of 446 strains isolated, 88 strains(19.73%) produced LT, while 224 strains(50.22%) produced ST, 134(30.05%) produced both LT-ST simultaneously. Serological typing were typed into 06, 08, 020, 025, 027, 0126, 0148 and 0159. Serotype 06 had the highest incidence of 26.46% followed by 0148(18.16%), 027(14.57%); 025(10.99%), 0159(4.03%), 0126(3.59%), 020(1.12%) and 08(0.45%). 2. Serotype 06, 08 and 025 almost always produced both LT and ST, whereas serotypes 020, 027 and 0148 almost always produced only ST. And serotypes 06, 025, 0126 and 0159 almost always produced LT or ST. 3. Of 134 LT-ST positive strains, 115 strains were serotype 06, 3 strains were 025, 2 strains were 08, and 14 strains were 0 untypable. 4. Of 224 ST positive strains, 65 strains were serotype 027, 81 strains were 0148, 16 strains were 0159, and 42 strains were 0 untypable. 5. Of 88 LT positive strains, 45 strains were serotype 025, 5 strains were 0126, 2 strains were 0159, and 42 strains were 0 untypable.

• Korean Journal of Veterinary Research
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• v.26 no.1
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• pp.69-77
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• 1986

The purpose of this study was the examination for presence of K99 antigen (K99), enterotoxigenicity, 0-groups, colicin and antibiotic susceptibility among E. coil isolated from calves and cows. A total of 49(18.7%) among 262 strains, isolated from 30(26.5%) out of 113 calves and cows, possesed K99, and thirty three of 49 $K99^+$ strains produced ST. Of the strains of diarrheal calf origin which less than 15 days old, a high correlation was observed between enterotoxigenic ability and K99: 92.3% of the $K99^+$ strains produced heat stable enterotoxin(ST). In O group typing of 33 $ST^+$ strains, they belonged to O20(48.4%), O8(9.1%), O9(6.1%), O139(6.1%), O149(6.1%), O101(3.0%), O115(3.0%), except six which were untypable or autoagglutinable. Of 262 E. coil isolates, 30 strains(13.3%) produced colicin and K99 were detected in 6 strains. One hundred eighty eight strains(71.8%) of 262 E. coil isolates were resistance to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, tetracycline, alone or in combination thereof. One hundred and fourteen(60.6%) out of 188 drug resistance strains carried R factor($R^+$) which were transferable to the recipients by conjugation. Sixty five $R^+$ strains(57.0%) carried thermo-sensitive R plasmid.

• The Journal of the Korean Society for Microbiology
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• v.22 no.2
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• pp.139-145
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• 1987

Adherence to HEp-2 cells has been proposed as a virulence characteristic of enteropathogenic E. coli (EPEC). The role of the HEp-2 adherent E. coli was evaluated in a group of children with endemic diarrhea admitted to Hanyang University Hospital in Seoul, Korea. HEp-2-adherent E. coli was detected in fecal samples of 59 (59%) of 100 cases and ten (22.7%) of 44 concurrent control children (p<0.0005). Adherence was exhibited by 15 serogroups and subgroups, but within these groups more than one adherence pattern was frequently observed. Of 17 strains belonging to traditional infantile EPEC serogroups, 12(70.6%) gave a positive adherence. Of 45 enterotoxin producing strains, 24 (53.3%) gave a positive adherence. HEp-2-adherent strains that did not belong to classic EPEC serogroups and did not produce heat-stable and/or heat-labile enterotoxins(referred as enteroadherent E. coli, EAEC) was found in 29 (29%) of the patients with diarrhea and in six (13.6%) of the well children (p<0.05). From 22 of the 29 cases, no pathogen other than EAEC was isolated. These findings strongly implicate EAEC as the cause of diarrhea in the children. Our study supports the concept that EAEC may be an important cause of endemic diarrhea in Korean children.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.286-294
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• 2004

Between 1997 and 1998 in Korea, 56 isolates of Escherichia coli were obtained from pig suffering diarrhea. Among those, 38 isolates that showed the hemolytic activity, antimicrobial resistance, and toxin production were studied. Among 38 isolates, thirty-six isolates $(94.7\%)$ were resistant to tetracycline, 27 isolates $(71.0\%)$ were resistant to ampicillin, 26 isolates $(68.4\%)$ were resistant to chloramphenicol, and 21 isolates $(55.2\%)$ were resistant to trimethoprim, while none was resistant to aztreonam, amikacin, and norfloxacin. Among these iso­lates, 21 isolates $(55.3\%)$ were multiple drug resistant to at least four different class antimicrobial agents. Extended spectrum $\beta-lactamase$ producing isolates were not detected in the double disk synergy test. In these hemolytic Escherichia coli, heat-stable enterotoxin $(89.5\%)$ was the most prevalent toxin, followed by vero­toxins $(47.4\%),$ and then heat-labile enterotoxin $(31.6\%).$ Except 8 isolates $(21.0\%)$ which produced ST only, 12 isolates $(31.6\%)$ produced ST and LT, 13 isolates $(34.2\%)$ produced ST, VT, and VTe, and 5 isolates $(13.2\%)$ produced VT and VTe. However, none produced all 4 types of toxin, simultaneously. The predominant serotype could not be determined by the agglutination method. Sixteen isolates $(42.1\%)$ were strongly adhered to T-24 bladder cell and 17 isolates $(44.7\%)$ were to Caco-2 intestinal cell. Especially, 11 strains $(28.9\%)$ were evaluated as strongly adhesive to both T-24 cells and Caco-2 cells. Genes for toxin and the antimicrobial resistance were transferred to clinical isolates of Escherichia coli from human urine by the filter mating method. Results suggest the possibility that antimicrobial resistance and toxin can be transferred from animals to humans by direct con­tact of resistant bacteria as well as gene transfer, although there was no correlation between toxin production, adherent activity, and antimicrobial resistance among hemolytic E. coli isolated from pig suffering diarrhea.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.262-270
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• 2017

Yersinia enterocolitica is a well-known foodborne pathogen causing gastrointestinal infections worldwide. The strain Y. enterocolitica FORC_002 was isolated from the gill of flatfish (plaice) and its genome was sequenced. The genomic DNA consists of 4,837,317 bp with a GC content of 47.1%, and is predicted to contain 4,221 open reading frames, 81 tRNA genes, and 26 rRNA genes. Interestingly, genomic analysis revealed pathogenesis and host immune evasion-associated genes encoding guanylate cyclase (Yst), invasin (Ail and Inv), outer membrane protein (Yops), autotransporter adhesin A (YadA), RTX-like toxins, and a type III secretion system. In particular, guanylate cyclase is a heat-stable enterotoxin causing Yersinia-associated diarrhea, and RTX-like toxins are responsible for attachment to integrin on the target cell for cytotoxic action. This genome can be used to identify virulence factors that can be applied for the development of novel biomarkers for the rapid detection of this pathogen in foods.

• The Journal of the Korean Society for Microbiology
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• v.22 no.2
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• pp.147-153
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• 1987

Colonization factor antigen I(CFA I) has been shown to be one of several virulence factors that promote attachment of enterotoxigenic E. coli(ETEC) to small intestinal epithelial cells of humans. The ability of ETEC to produce mannose-resistant hemagglutination(MRHA) of human blood group A has been used to detect CFA I. To determine gastrointestinal carriage in Korean children of E. coli with MRHA and CFA I, 116 strains of E. coli from diarrheal children admitted to Hanyang University Hospital were examined for MRHA of human erythrocytes and the presence of CFA I. Of 45 ETEC strains, 18(40%) gave a positive MRHA($MRHA^+$) and eight(18%) were positive for CFA I(CFA $I^+$). ETEC with CFA I were all heat-stable enterotoxin(ST) producers and two of these strains were of serogroups $O_{25}$. Of 17 classic enteropathogenic E. coli(EPEC), 7(41%) were $MRHA^+$ but all were negative for CFA I(CFA $I^-$). Of 30 enteroadherent E. coli(EAEC) strains, 11(37%) were $MRHA^+$ and one was CFA $I^+$. Of 24 nonpathogenic E. coli, 4(17%) were $MRHA^+$ but all were CFA $I^-$. It was shown that MRHA was common in all strains of E. coli, CFA I was limited only to ST producing ETEC and EAEC; although MRHA is a useful screening procedure, serologic tests seem to be necessary to comfirm CFA I production. CFA I was associated with a lower proportion of ETEC isolates in Korea than has been reported for other locations.