• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.3
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• pp.571-575
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• 2004

Escherichia coli O157:H7 cause hemorrhagic colitis and the extraintestinal complication of hemolytic-uremic syndrome, with their higher incidence occurring in children. Lipopolysaccharide (LPS) of E. coli O157:H7 is very important to make IgG anti-LPS with bactericidal activity. To identify the characteristic of E. coli OI57:H7, we isolated 60 MDa plasmid and amplified stx genes of shiga-like toxin (Stx) 1, 2 of E. coli O157:H7 by polymerase chain reaction (PCR) method. Using the simple purification method which contained phenol extract, ethanol precipitation and gel filtration steps, the LPS of E. coli O157:H7 was isolated and purified. Finally, we confirmed the purity of LPS through SDS-PAGE and silver nitrate staining.

• Clinical and Experimental Pediatrics
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• v.57 no.2
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• pp.96-99
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• 2014

Hemolytic uremic syndrome (HUS) is one of the most common causes of acute renal failure in childhood and is primarily diagnosed in up to 4.5% of children who undergo chronic renal replacement therapy. Escherichia coli serotype O157:H7 is the predominant bacterial strain identified in patients with HUS; more than 100 types of Shiga toxin-producing enterohemorrhagic E. coli (EHEC) subtypes have also been isolated. The typical HUS manifestations are microangiopathic hemolytic anemia, thrombocytopenia, and renal insufficiency. In typical HUS cases, more serious EHEC manifestations include severe hemorrhagic colitis, bowel necrosis and perforation, rectal prolapse, peritonitis, and intussusceptions. Colonic perforation, which has an incidence of 1%-2%, can be a fatal complication. In this study, we report a typical Shiga toxin-associated HUS case complicated by small intestinal perforation with refractory peritonitis that was possibly because of ischemic enteritis. Although the degree of renal damage is the main concern in HUS, extrarenal complications should also be considered in severe cases, as presented in our case.

• Korean Journal of Agricultural Science
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• v.42 no.3
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• pp.201-205
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• 2015

The experiment was conducted to evaluate the effects of pathogenic Escherichia coli on diarrhea, growth performance, and blood profile of weaned pigs. A total of 48 pigs were used and housed in individual pens of disease containment chambers for 16 d (4 d before and 12 d after the first challenge [d0]). The treatments were with or without the pathogenic E. coli challenge (F-18 E. coli strain; heat-labile, heat-stable, and Shiga-like toxins). Pigs were orally inoculated with a dose of $10^{10}cfu$ E. coli per 3 mL PBS daily for 3 days. The common nursery diet and water were available at all times. The ADG, ADFI, G:F, diarrhea score, ratio of fecal ${\beta}$-hemolytic coliforms from total coliforms (RHT), and blood profile were measured. The pathogenic E. coli reduced (P < 0.05) ADG from d0 to 6 (117 vs. 297 g/d) and from d0 to 12 (377 vs. 238 g/d) compared with the control. Meanwhile, the pathogenic E. coli increased (P < 0.05) diarrhea score (average 3.4 vs. 1.4) and RHT (average 82 vs. 11%) on d3, 6, and 9 and the number of white blood cells (17.59 vs. $13.48{\times}10^3/{\mu}L$) on d6 compared with the control. No differences were found on ADFI and others in the blood profile (total protein and hematocrit). In conclusion, pathogenic E. coli used in this experiment successfully caused mild diarrhea, increased number of white blood cells, and adversely affected growth rate of weaned pigs.

• BMB Reports
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• v.37 no.5
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• pp.574-581
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• 2004

The full-length cDNA of the lumbrokinase fraction 6 (F6) protease gene of Lumbricus rubellus was amplified using an mRNA template, sequenced and expressed in E. coli cells. The F6 protease gene consisted of pro- and mature sequences by gene sequence analysis, and the protease was translated and modified into active mature polypeptide by N-terminal amino acid sequence analysis of the F6 protease. The pro-region of F6 protease consisted of the 44 residues from methionine-1 to lysine-44, and the mature polypeptide sequence (239 amino acid residues and one stop codon; 720 bp) started from isoleucine-45 and continued to the terminal residue. F6 protease gene clones having pro-mature sequence and mature sequence produced inclusion bodies in E. coli cells. When inclusion bodies were orally administrated rats, generated thrombus weight in the rat' venous was reduced by approximately 60% versus controls. When the inclusion bodies were solubilized in pepsin and/or trypsin solutions, the solubilized enzymes showed hemolytic activity in vitro. It was concluded the F6 protease has hemolytic activity, and that it is composed of pro- and mature regions.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.519-527
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• 2021

In this study, the hemolysis of Enterococcus faecalis BMSE-HMP strains, isolated from human breast milk, was investigated, and the anti-hemolytic and antimicrobial effects on multidrug-resistant (MDR) bacteria were investigated. The enzyme activity of E. faecalis BMSE-HMP 4 strains was measured, and it was found that the activities of esterase and esterase lipase were the highest. In addition, no hemolytic reaction was observed in any of the isolates. Subsequently, the anti-hemolytic activity against MDR strains causing hemolysis was evaluated. E. faecalis BMSE-HMP002 had the highest anti-hemolytic activity against Staphylococcus aureus CCARM 3855 at 75.71 ± 10.00%. The anti-hemolytic activity against Escherichia coli DC 2 CCARM 0238 and Pseudomonas aeruginosa CCARM 0223 showed that the activity of BMSE-HMP001 was highest at 76.92 ± 2.99% and 87.93 ± 1.93%, respectively. Examination of the antimicrobial effects against the MDR bacteria Staphylococcus spp., Escherichia spp., Pseudomonas spp., Salmonella spp., Klebsiella spp., Enterobacter spp., and E. faecalis BMSE-HMP strains showed antimicrobial effects against both gram-positive and gram-negative strains. Breastfeeding delivers enterococci into the intestinal tract of newborns by lactation, and its usefulness is attracting attention as it has been reported that enterococci have a potential effect on neonatal immune development. In this study, the hemolytic and antimicrobial effects of E. faecalis BMSE-HMP strains on MDR bacteria were investigated, to confirm their potential as useful lactic acid bacteria. Additional studies on the antibiotic resistance and toxicity of the E. faecalis BMSE-HMP strains, isolated in this study, are necessary to prove it safe for use.

• Pediatric Infection and Vaccine
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• v.4 no.1
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• pp.73-78
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• 1997

Purpose : Escherichia coli O157 can produce diarrhea as well as hemorrhagic colitis and hemolytic uremic syndrome. In many parts of North America, E. coli O157 often is the second or third most commonly isolated enteric bacterial pathogens. Recently, intakes of fast food, including hamburgers have increased in Korea. Therefore, E. coli O157 infection in Korea are likely to be increased. Methods : Stool samples from 317 pediatric diarrheal patients were analyzed by culture on sorbitol-MacConkey agar. Sorbitol-negative colonies were teated by E. coli O157 latex agglutination test. Results : Of the 317 specimens, one (0.3%) were E. coli O157:NM that not produced Shiga toxin. The 7 year old male patient who had complained of abdominal pain, vomiting and non-bloody diarrhea for 2 days. The patient was improved for 2 days after admission. Conclusions 1 Only one (0.3%) of all fecal samples were isolated E. coli O157 that not produced Shiga toxin. Therefore, routine stool culture for the isolation of E. coli O157 was not likely to be neccessary so far.

• 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.

• Korean Journal of Veterinary Research
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• v.45 no.1
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• pp.99-103
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• 2005

We report septicemic colibacillosis accompanied by white-spotted kidney in a 30-day old female Holstein-Friesian dairy calf. Grossly, there were numerous white spots sized average 0.5cm in diameter in both kidneys. When sectioned sagittally, there were radially oriented gray streaks extending outward and reaching the renal cortex. The renal papillae were ulcerated, white to gray in color and very friable. Histologically, there was extensive purulent inflammation characterized by severe neutrophilic cellular infiltrations in the tubular lumens and interstitia. In addition, massive coagulative necrosis were found in the apices of papillae. Numerous Gram-negative bacterial colonies were detected in both of the renal and lung tissues. Beta-hemolytic Escherichia coli (E. coli) was purely isolated from the renal parenchyma, peritoneal surface and pleural surface. Based on the above results, we suggest that the generalized septicemia with pyelonephritis may be oriented from the lower urinary tract infection with E. coli in this case.

• Korean Journal of Veterinary Service
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• v.39 no.2
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• pp.87-92
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• 2016

Porcine edema disease (ED) is a communicable disease of pigs caused by infection with Shiga toxin (Stx)-producing Escherichia coli (STEC) which expresses F18 fimbriae and/or Stx type 2e (Stx2e). While STEC causes a severe illness including hemorrhagic colitis and hemolytic-uremic syndrome in humans, it induces damage to the vascular endothelium, which results in edema, hemorrhage, and microthrombosis, leading in high mortality in pigs. In the present study, we cultured Stx2e-producing E. coli field isolates from conventional pig farms that experienced sudden deaths previously with symptoms similar to porcine edema disease, which were further investigated with Shiga toxin profiles. A total of 43 strains were identified from the collected samples by F18 or Stx2e specific PCR. Based on the PCR, 42 isolates out of 43 isolates were proved to carry one of F18 or Stx2e genes and 14 isolates to carry both F18 and Stx2e genes. All of the 30 isolates that harbored Stx2e gene induced the cytopathic effect (CPE) in vero cells and especially, the isolate 150229 produced the highest level of Shiga toxin. Therefore, we identified the virulence genes (F18 and Stx2e) and demonstrated Shiga toxin-producing abilities from porcine edema disease causing E. coli filed isolates. These results suggested that one of the isolates could be a vaccine antigen candidate against STEC through further investigating to elicit an immune response.

• Korean Journal of Food Science and Technology
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• v.47 no.1
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• pp.126-131
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• 2015

Pathogenic Escherichia coli is recognized as an important cause of diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome worldwide. This study was conducted to investigate the prevalence E. coli contamination in the Korean traditional food bibimbap. E. coli were isolated from 84 of 1142 (7.3%) bibimbap investigated from 2005 to 2011. Antibiotic resistance profiling demonstrated that 6 of the 84 isolates (7.2%) showed multiple drug resistance. Fifteen virulence genes specific for pathogenic E. coli such as Shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and enteroaggregative E. coli (EAEC) were examined by multiplex PCR for mixed bacterial cultures derived from bibimbap samples. The EPEC virulence gene (ent) was detected in 5 strains (5.9%), while ETEC, EAEC, and EIEC were not detected. STEC serotypes O103 (1.2%), O91 (1.2%), and O128 (6.0%) were found, but other serogroups such as O26, O157, O145, O111 and O121 were not detecded. Automated Repetitive-Sequence-Based PCR analysis showed different patterns.