• Title/Summary/Keyword: E. coli K-12

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Modified sorbitol MacConkey agar for the rapid isolation of Escherichia coli O157:H7

  • Jung, Byeong-yeal;Jung, Suk-chan;Lee, Na-kyung;Cho, Seong-kun;Cho, Dong-hee;Her, Moon;Yoon, Yong-dhuk;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.765-771
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    • 1999
  • Unlike most Escherichia coli strains, E coli O157 : H7 didn't ferment sorbitol within 24h of incubation and showed a negative reaction for $\beta$-glucuronidase. We developed a new medium for the rapid isolation of E coli O157 : H7 using sorbitol MacConkey agar with cefixime, potassium tellurite and 4-methylumbelliferyl-${\beta}$-D-glucuronide (MUG) as a primary plating medium. The addition of $20{\mu}g/ml$ of vancomycin in enrichment broth for E coli O157 : H7 inhibited lots of Gram positive bacteria. Three strains (10.3%) of 29 non-O157 E coli strains and 3 strains (8.3%) of 36 Salmonella spp were inhibited at the $0.05{\mu}g/ml$ of cefixime and 23 strains (79.3%) of 29 non-O157 E coli strains and 12 strains (33.3%) of 36 Salmonella spp were inhibited at the $2.0{\mu}g/ml$ of potassium tellurite. But none of the E coli O157 : H7 was affected at these concentration. The addition of MUG at $100{\mu}g/ml$ level to sorbitol MacConkey agar with cefixime and potassium tellurite (CTM-SMAC) aided in the rapid isolation of E coli O157 : H7 from samples by checking sorbitol-negative and $\beta$-glucuronidase negative phenotypes simultaneously. In conclusion, inoculation of a positive in the O157 screening test from enrichment broth on CTM-SMAC appeared to be a rapid, cost-effective and sensitive method for the isolation of E coli O157 : H7.

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Photo or Solar Ferrioxalate Disinfection Technology without External Hydrogen Peroxide Supply

  • Cho, Min;Jeong, Joon-Seon;Kim, Jae-Eun;Yoon, Je-Yong
    • Environmental Engineering Research
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    • v.12 no.5
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    • pp.238-243
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    • 2007
  • The Fenton reaction, which refers to the reaction between ferrous ions and hydrogen peroxide to produce the OH radical, has not been widely applied to the disinfection of microorganisms despite being economic and environmentally friendly. Cho et al. have previously proposed the neutral photo ferrioxalate system as a solution to the problems posed by the Fenton reaction in acidic conditions, but this system still requires an external hydrogen peroxide supply. In the present study, we developed a simple disinfection technology using the photo or solar ferrioxalate reaction without the need for an external hydrogen peroxide supply. E. coli was employed as the indicating microorganism. The study results demonstrated the effectiveness of the photo ferrioxalate system in inactivating E. coli without any external hydrogen peroxide supply, as long as dissolved oxygen is supplied. Furthermore, the solar ferrioxalate system achieved faster inactivation of E. coli than an artificial light source at similar irradiance.

Detection of Escherichia coli O157:H7 Using Immunosensor Based on Surface Plasmon Resonance

  • Oh, Byung-Keun;Kim, Young-Kee;Bae, Young-Min;Lee, Won-Hong;Choi, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.780-786
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    • 2002
  • An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

Evaluation of Epidemic Characteristics of Extended Spectrum β-Lactamase Producing Bacteria Isolated from Blood Cultures (혈액배양에서 분리된 Extended Spectrum β-Lactamase 생성균의 역학적 특성 조사)

  • Seo, Choong-Won;Kim, Sang-Ha;Hwang, Seock-Yeon;Kim, Young-Kwon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.9
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    • pp.2516-2522
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    • 2009
  • The data which analyze the results of blood cultures are crucial basic information of the empirical therapy for patients with infection since the patterns of the species of microorganism isolated from blood and the results of antibiotic susceptibility test vary depending upon patients general features. Especially, in case of ESBL-producing bacteria, there is a close relation with use of antibiotics. Therefore, we carried out the research with the results of blood culture and antibiotic. 1. Total 39,305 cases of blood culture samples were investigated and positive patients of 2,216 (20.0%) were found. Among those, there were 40 patients with ESBL positive, and blood culture positive samples were 4,798 (12.2%). ESBL positive bacteria were found in 86 samples (including double checked culture bacteria). 2. The majority of ESBL producing bacteria were E. coli, K. pneumoniae and K. oxitoca as ordering based on the number. 3. The research showed the results that there were more females than male with the bacterias, more E. coli in over 50 years old aged group than other bacterias, more K. pneumoniae and K.oxitoca in less 1 year old aged group than other bacterias and largest numbers of patients with 13 patients (32.5%) in Chungcheongnam-do province were found. 4. The most common ESBL producing bacteria were E. coli throughout 3 years, but K, pneumoniae and K. oxitoca were also fairly found. Interestingly, E. coli was highly found in over 50 years old patients.

Prevalence of pili and enterotoxins of Escherichia coli associated with diarrhea in preweaning piglets (포유자돈 소장에서 분리된 대장균의 섬모항원과 장내독소 분포양상)

  • Ham, Hee-jin;Cheon, Doo-sung;Chae, Chan-hee
    • Korean Journal of Veterinary Research
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    • v.37 no.4
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    • pp.779-784
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    • 1997
  • A comprehensive study of 132 Escherichia coli isolates from 150 piglets with colibacillosis included detection of heat-labile enterotoxin, heat-stable enterotoxin, and identification of K88 (F4), K99 (F5), 987P (F6), and F41. Four pili were examined by haemagglutination and slide agglutination test. Heat-labile(LT) and heat-stable(ST) enterotoxin was determined by reverse passive latex agglutination and precipitation test, respectively. Among 132 E coli isolates, 26 had K88 (19.7%), 16 had K99 (12.1%), 3 had 987P (2.3%), and 2 had F41 (1.5%). Three had K88 and K99 (2.3%), 3 had K88 and 987P (2.3%), 2 had K99 and 987P (1.5%), 5 had K99 and F41 (3.8%), and 8 E coli strains had K88, K99 and F41 (6.1%) simultaneously. Among 132 E coli isolates, 5 produced LT only (3.8%), 55 produced heat-stable toxin ST only (41.7%), and 4 produced both LT and ST (3.0%). Three major pathotypes accounted for 27.9% of E coli isolates: $K99^+$ (8.3%), $K88^+ST^+$ (9%) and $K88^+$ (10.6%). Results of this study indicated that piliated enterotoxin-producing E coli was prevalent and was associated with diarrhea in preweaning piglets.

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Characterization and Cloning of a Phytase from Escherichia coli WC7. (Escherichia coli WC7가 생산하는 Phytase의 효소특성과 그 유전자의 클로닝)

  • 최원찬;오병철;김형권;강선철;오태광
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.1-7
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    • 2002
  • Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

Susceptibility of Escherichia coli Isolated from Cattle to Some Antimicrobial Agents (소에서 분리(分離)한 Escherichia coli의 항생물질내성(抗生物質耐性) 및 전달성내성인자(傳達性耐性因子)의 분포(分布))

  • Park, Cheong Kyu
    • Korean Journal of Veterinary Research
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    • v.17 no.1
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    • pp.5-8
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    • 1977
  • One hundred and fifty seven Escherichia coli strains isolated from 18 cattle (9 dairy cattle received penicillin, streptomycin (SM) or sulfadimethoxine for treatment of diseases and 9 Korean native cattle not received antibiotics) were studied for the drug resistance and distribution of R factors. Of 88 E. coli strains isolated from cattle not received antibiotics, only 1 strain was resistant to SM, but about 46 per cent of 69 E. coli strains isolated from cattle received antibiotics were resistant to SM, tetracycline (TC), ampicillin (AP), kanamycin (KM), chloramphenicol (CM), and sulfisomidine (Su), alone or in combination thereof. Of resistant strains, about 72% were resistant to three or more antibiotics, but 28% were found to singly resistant. The most frequent resistant pattern was triple resistance to AP, KM and Su (37.6%), and quadruple one to SM, TC, CM and Su (12.5%). About 28% of resistant strains carried R factors which were transferable to E. coli ML 1410 $NA^r$ by conjugation.

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Antimicrobial Resistance and Multi-Drug Resistance Patterns of Pathogenic Bacteria Isolated from Food Poisoning Patients in Incheon (인천지역 식중독 환자에서 분리한 병원성 세균의 항생제 내성 및 다제 내성 양상)

  • Huh, Myung-Je;Oh, Sung-Suck;Jang, Jae-Seon
    • The Korean Journal of Food And Nutrition
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    • v.26 no.1
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    • pp.132-136
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    • 2013
  • Antimicrobial resistance and multi-drug resistance patterns have been carried out on total of 210 isolated of Salmonella spp. and pathogenic E. coli isolated from food poisoning patients on January through December 2012 in Incheon, Korea. The highest percentage of antibiotics resistance was found to the following antimicrobial agents: tetracycline 43.8%, ampicillin 34.8%, nalidixic acid 23.8%, sulfamethoxazole/trimethoprim and chloramphenicol 12.4%, and ampicillin/sulbactam 11.4%. The highest percentage of resistance was 37.5% to ampicillin for Salmonella spp. and 59.0% to tetracycline for pathogenic E. coli. Overall the multidrug resistance rates of 1 drug was 26.2%, 2 drugs 9.0%, 3 drugs 9.5%, 4 drugs 7.1%, and 5 or more drugs 12.46%. The multi-drug (MDR) strains to four or more antimicrobial agents among the resistant organisms were quite high: 15.9% and 22.1% for Salmonella spp. and pathogenic E. coli, respectively. The study implies that limitation of unnecessary medication use is pertinent in order to maintaining the efficacy of drugs.

Expression of E. coli Phosphofructokinase Gene in an Autotrophic Bacterium Acidithiobacillus thiooxidans

  • Tian, Ke-Li;Lin, Jian-Qun;Liu, Xiang-Mei;Liu, Ying;Zhang, Chang-Kai;Yan, Wang-Ming
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.56-61
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    • 2004
  • A plasmid pSDK-l containing the Escherichia coli phosphofructokinase-l gene (pfkA) was constructed, and transferred into extremely acidophilic Acidithiobacillus thiooxidans Tt-7 by conjugation with the aid of plasmid RP4 at a frequency of $10^{-5}$ per recipient. This plasmid was stable in A. thiooxidans. The pfkA gene from E. coli could be expressed in this obligately autotrophic bacterium, but the enzyme activity (21.6 U/g protein) was lower than that in E. coli (K12: 85.9 Dig protein; DF1010 carrying plasmid pSDK-l: 96.6 U/g protein). In the presence of glucose, the Tt-7 transconjugants consumed glucose, leading to a better growth yield.

Improvement of 4-chlorobiphenyl degradation bya recombinant strain, pseudomonas sp. DJ12-C

  • Kim, Ji-Young;Kim, Young-Chang;You, Lim-Jai;Lee, Ki-Sung;Ok, Ka-Jong;Hee, Min-Kyung;Kim, Chi-Kyung
    • Journal of Microbiology
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    • v.35 no.1
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    • pp.53-60
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    • 1997
  • Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 isolated from the polluted environment are capable of degrading biphenyl and 4-chlorobiphenyl (4CB) to produce benzoic acid and 4-chlorobenzoic acid (4CBA) respectively, by pcbABCD-encoded enzymes. 4CBA can be further degraded by Pseudomonas sp. DJ-12, but not by Pseudomonas sp P20. However, the meta-cleavage activities of 2, 3-dihydroxybiphenyl (2, 3-DHBP) and 4-chloro-2, 3-DHBP dioxygenases (2, 3-DHBD) encoded by pcbC in Pseudomonas sp. P20 were stronger than Pseudomonas sp. DJ-12. In this study, the pcbC gene encoding 2, 3-DHBD was cloned from the genomic DNA of Pseudomonas sp. P20 by using pKT230. A hybrid plasmid pKK1 was constructed and E. coli KK1 transformant was selected by transforming the pKK1 hybrid plasmid carrying pcbC into E. coli XL1-Blue. By transferring the pKK1 plasmide of E. coli KK1 into Pseudomonas sp. DJ-12 by conjugation, a recombinant strain Pseudomonas sp. P20, Pseudomonas sp. DJ-12, and the recombinant cell assay methods. Pseudomonas sp. DJ12-C readily degraded 4CB and 2, 3-DHBP to produce 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA), and the resulting 4CBA and benzoic acid were continuously catabolized. Pseudomonas sp. DJ12-C degraded 1 mM 4CB completely after incubation for 20 h, but Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 showed only 90% and Pseudomonas sp. DJ-12 had, but its degradation activity to 2, 3-DHBP, 3-methylcatechol, and catechol was improved.

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