• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.25 no.1
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• pp.79-86
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• 2022

Purpose: To determine the effect of zinc deficiency on fecal protein, electrolyte, and short-chain fatty acid levels in both heat-stable (ST) and heat-labile (LT) enterotoxigenic Escherichia coli (ETEC)-induced diarrhea in rats. Methods: Albino rats, weighing 100 to 150 g, were divided into 2 groups, with 15 animals each: non-zinc and zinc-deficient. These two groups were sub-divided into three sub-groups with five rats each: control (saline); LT-ETEC; and ST-ETEC. Sodium phytate (30 mmol/L) was added to the animals' water to induce zinc deficiency, while diarrhea was induced using 5×10⁹ ETEC cells/mL. Fecal protein levels were estimated using the Bradford method, while sodium and potassium levels were determined using atomic absorption spectrophotometry. Short-chain fatty acids were measured using gas chromatography-mass spectrometry. Results: Among the non-zinc and zinc-deficient groups, there were significant increases (p=0.04), (p=0.03) in fecal protein concentrations (mg/mL) in the LT-ETEC- (4.50±0.33), (6.50±0.26) and ST-ETEC- (3.85±0.19), (5.98±0.32) induced groups compared to the control groups (2.60±0.52), (3.50±0.11) respectively. Fecal sodium and potassium levels (mg/L) were significantly (p=0.029) increased in non-zinc-deficient rats induced with LT-ETEC (9.35±0.95, 1.05±0.48), and ST-ETEC (9.96±1.02, 1.21±0.45) compared with the control group (8.07±0.44, 0.47±0.17) but the increase were not statistically significant (p=0.059) in the zinc deficient rat groups. Fecal acetate and propionate levels (mg/g) significantly (p=0.032) increased when induced with LT-ETEC and ST-ETEC in non-zinc and zinc-deficient groups compared with the control groups. Conclusion: Zinc deficiency among rats with ETEC-induced diarrhea elevated fecal protein loss but may not have an effect on fecal sodium, potassium and short-chain fatty acid levels.

• Journal of Microbiology
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• v.43 no.4
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• pp.354-360
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• 2005

Heat-labile enterotoxin B subunit (LTB) of enterotoxigenic Escherichia coli (ETEC) is both a strong mucosal adjuvant and immunogen. It is a subunit vaccine candidate to be used against ETEC-induced diarrhea. It has already been expressed in several bacterial and plant systems. In order to construct yeast expressing vector for the LTB protein, the eltB gene encoding LTB was amplified from a human origin enterotoxigenic E. coli DNA by PCR. The expression plasmid pLTB83 was constructed by inserting the eltB gene into the pYES2 shuttle vector immediately downstream of the GAL1 promoter. The recombinant vector was transformed into S. cerevisiae and was then induced by galactose. The LTB protein was detected in the total soluble protein of the yeast by SDS-PAGE analysis. Quantitative ELISA showed that the maximum amount of LTB protein expressed in the yeast was approximately $1.9\%$ of the total soluble protein. Immunoblotting analysis showed the yeast-derived LTB protein was antigenically indistinguishable from bacterial LTB protein. Since the whole-recombinant yeast has been introduced as a new vaccine formulation the expression of LTB in S. cerevisiae can offer an inexpensive yet effective strategy to protect against ETEC, especially in developing countries where it is needed most.

• Korean Journal of Veterinary Service
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• v.43 no.2
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• pp.53-58
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• 2020

Enterovirulent Escherichia coli are among the most important causes of diarrhea in cattles. Between January and December, 2017, a total of 150 stool specimens from cattles were investigated for enterohaemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enterotoxigenic E. coli (ETEC) and enteroinvasive E. coli (EIEC) using real-time PCR. 131 E. coli were isolated from feces. The most frequently isolated pathotype in feces was EHEC (37 isolates). EPEC, ETEC and EAEC were detected in feces with 14, 7 and 3 respectively. EIEC was not detected. Antimicrobial resistance test was performed by agar disc diffusion method with 14 antimicrobials. Enterovirulent E. coli isolates showed the highest antimicrobial resistance to ampicillin 61.3%, followed by tetracycline 54.5% and streptomycin 45.5%. They had low resistance to amikacin 11.4%. Of 44 isolates, 37 (84.1%) were resistant to more than 2 antimicrobials. futher study a highest antimicrobial susceptibility to trimethoprim/sulfamethoxazole 50.0% and florofenicol 47.7%.

• The Journal of the Korean Society for Microbiology
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• v.22 no.2
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• pp.155-162
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• 1987

To determine the prevalence of antibiotic resistance in fecal E. coli and to investigate possible associations between antibiotic resistance and other plasmid-mediated virulence properties, antibiotic disk susceptibility tests for nine antibiotics were done on 141 strains of E. coli isolated from diarrheal children and well controls. Eighty two percent of the test strains were resistant to one or more antibiotics. Antibiotics to which the test strains were most resistant in descending order were ampicillin (85%), trimethoprim/sulfamethoxazol (60%), and cephalothin (55%). Seventy nine percent of these resistant strains were resistant to two or more antibiotics. All 141 test strains were sorted into enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroadherent E. coli (EAEC) and non-pathogenic E. coli and the percentages of strains resistant to multiple antibiotics were compared. Among ETEC regardless of its source, multiple drug resistance was more frequent in strains producing heatstable enterotoxin (ST) only than in strains producing only heat-labile enterotoxin (LT) or both. In EAEC, multiple resistance was more frequently associated with strains isolated from diarrheal patients than with those from well controls. The major antibiotic resistance patterns possessed by multiple resistant enteropathogenic strains were $SXT^R$ $AM^R$, $CR^R$, and $SXT^R$ $AM^R$ $CR^R$. Of 28 ST- producing $SXT^R$ ETEC, 26(96%) were also resistant to ampicillin and 17 (61%) were resistant to cephalothin. The similar pattern was observed in EAEC and EPEC as well. This study has important implications for the treatment of E. coli diarrhea with antibiotics because it is possible that dissemination of virulence could occur under the force of selective antibiotic pressure. In addition, this study suggests that the in vivo efficacy of SXT in treating diarrheal illness be reevaluated.

• 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)

• Korean Journal of Veterinary Service
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• v.20 no.4
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• pp.359-370
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• 1997

This study was carried out to Investigate the biochemical characteristics, antibiotic susceptibility, serogroups and pili producibility test of enterotoxigenic Escherichia coli(ETEC) isolated from piglets with diarrhea in Kangwon province from March to October 1996. 1. Sixty eight E coli strains were isolated from 72 piglets with diarrhea and the biochemical and cultural reaction were compared with the classification criteria of Edwards and Ewing. 2. The serogroups of 26 isolates were classified as 08 : K87 6(8.8%), O20 : K1O1 4(5.9%), O141 : K85 4(5.9%), 09 : K103 : P987 3(4.4%), O45 : K 2(2.9%) 0139 : K82 2(2.9%), O64 : $K^{-}$2(2.9%), O149 : K91 1(1.5%), O157 : K88ac 1(1.5%) and O115 : $K^{-}$1(1.5%), respectively. 3. In antibiotic susceptibility test, the isolates showed high susceptible to Ak, Eno, Na, Gm, Am and Km, whereas resistance to Tc, Sm and Cf. 4. Sixty one strains(89.7%) of 68 I coli Isolates were resistant to one or more drugs. The isolates resistant to 2 and 3 or more drugs were 60.3% and 19.1%, respectively. Amog the 16 multiple resistant patterns, Sm Tc(11.5% ), Cf Sm Tc(11.5% ), Cf Cp Sm Su Tc(9.8% ) and Cf Cp Sm Su Tc(8.2%) patterns were frequently observed. 5. MRHA of guinea pig erythrocytes was detected in 9 out of 26 OK serotype and 9 out of 42 unidentified serotypes. MRHA titers of serotypes showed from 16 to 32 in O141 : K85 and no titers in O139 : K82. 6. By the GM1 ganglioside ELISA, $\beta$-, $\alpha$-, and $\gamma$-hemolysin producing strains was detected as 36, 6, and 5 from heat labile enterotoxin(LT) of 47 ETEC, respectively. The distribution of LT toxin from 112 isolates was showed $\beta$- hemolysin, 2 isolates $\alpha$-hemolysin and 3 isolates $\gamma$-hemol-ysin from 26 OK serotypes.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.6
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• pp.777-785
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• 2010

The effect of two doses of different sources of zinc, inorganic (zinc oxide) or chelated (zinc glutamate chelate), on morphology and turn-over of the small intestine was assessed in early-weaned pigs orally challenged with enterotoxigenic E. coli K88 (ETEC). Sixty pigs weaned at 21 days were assigned to one of the following 5 diets: control (C); C+Zinc oxide (ZnO), either a 200 or a 2,500 mg Zn/kg dose; or C+zinc chelate with glutamic acid (Glu-Zn), either a 200 or a 2,500 mg Zn/kg dose. On d 2, the pigs were orally inoculated with 1.5 ml of a $10^{10}$ CFU/ml E. coli K88ac O148 suspension. Zinc supplements did not improve the performance of the pigs, but on d 5 faecal excretion of ETEC was reduced, and this was mainly due to high zinc doses (p<0.05). The villous height in the duodenum was improved by the zinc supplements (p<0.01) whatever the source and the level, whereas no effect was seen in the other two tracts of small intestine. The diet did not affect apoptosis and mitosis counts, while ETEC-susceptible pigs had more mitotic cells in the villi than non-susceptible pigs, particularly in the jejunum (p<0.01). The duodenum had fewer mitotic cells in the villi (p<0.05) and in the crypts (p<0.01) and more apoptotic cells in the villi. High dietary doses of ZnO or Zn-Glutamate improve villous height of the duodenum, but not of the jejunum and the ileum, and do not affect the epithelial proliferative activity and apoptotic index of intestinal mucosa of early-weaned pigs orally challenged with ETEC.

• Journal of Microbiology
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• v.44 no.5
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• pp.548-555
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• 2006

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.421-426
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• 2014

To characterize the extended-spectrum beta-lactamases (ESBLs) in diarrheagenic Escherichia coli from Korea in 2008-2011, we screened seven enterotoxigenic E. coli (ETEC) and one enteroaggregative E. coli (EAEC) that produce ESBLs from a nationwide survey. All eight isolates produced CTX-M-type ESBLs, including CTX-M-12 (n = 4), CTX-M-14 (n = 2), and CTX-M-15 (n = 2). PCR-based replicon typing indicated that the $bla_{CTX-M-12}$ genes of four ETEC isolates were carried on a conjugative IncF plasmid, whereas the $bla_{CTX-M-14}$ of one EAEC was located on an IncK plasmid. This is the first report of the occurrence of $bla_{CTX-M}$ genes in clinical isolates of EAEC in Korea. The ESBL-producing isolates were shown to be different based on pulsed-field gel electrophoresis and multilocus sequence typing, whereas the four isolates with CTX-M-12 were clonally related. These observations raise an alarm for the spread of plasmid-mediated resistance to ESBL among diarrheagenic E. coli.