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Optimal culture conditions for production of Escherichia coli Adhesin protein coupled to Escherichia coli Heat Labile Enterotoxin A2B in Escherichia coli TB1.

  • Lee, Yong-Hwa;Rhee, Dong-Kwon;Pyo, Suhk-Neung
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.226.2-226.2
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    • 2003
  • The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause of urinary tract infection. To produce a possible vaccine antigen against urinary tract infection, the fimH gene was genetically linked to the Itxa2b gene, which was then cloned into the pMAL -p2E expression vector. The chimaeric construction of pMALfimH/Itxa2b was transformed into Escherichia coli TB1 and its N-terminal amino acid sequence was analyzed. (omitted)

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Incidence and Risk Factors for Extended-Spectrum ${\beta}-Lactamase-Producing$ Escherichia coli in Community-acquired Childhood Urinary Tract Infection (지역사회 획득 소아 요로 감염에서 Extended-Spectrum ${\beta}-Lactamase$ 생성)

  • Lee Jung-Won;Shin Jee-Sun;Seo Jeong-Wan;Lee Mi-Ae;Lee Seung-Joo
    • Childhood Kidney Diseases
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    • v.8 no.2
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    • pp.214-222
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    • 2004
  • Purpose: Appropriate antibiotic therapy is important in childhood urinary tract infection and the selection of anibiotics is based on antimicrobial sensitivity of Escherichia coli. Extended-Spectrum ${\beta}-Lactamase(ESBL)$ is an enzyme produced by gram-negative bacilli that has the ability to hydrolyse penicillins, broad-spectrum cephalosporin and monobactam. There have been many reports of outbreaks of hospital infection by ESBL-producing organism. However, community-acquired infection with ESBL-producing organism are rare. This study was performed to retrospectively identify the incidence, characteristics and risk factors of ESBL (+) E. coli in community-acquired childhood UTI. Methods: In 288 children admitted in Ewha Womans University Hospital with E. coli UTI from Mar 2001 to February 2003, ESBL was isolated. ESBL was confirmed by the utilization of an automatized machine(Vitek GNS 433 card) using liquid medium dilution method according to National Committee for Clinical Laboratory Standard. The clinical characteristics, risk factors, antimicrobial resistance and treatment effectiveness were compared with ESBL(-) E. coli UTI. Results: Of 288 E. coli isolates, 31(10.8%) produced ESBL and 93.5%(29/31) occurred in infants younger than 6 month of age(P<0.01). No significant differences were noted in prior antibiotic use, prior admission history and underlying urogenital anomaly. Antimicrobial resistance was significantly higher in ESBL(+) E. coli compared with control patients (P<0.05). Although ceftriaxone showed 100% resistance in ESBL(+) E. coli, bacteriologic sterilization rate after ceftriaxone therapy was higher(96.8%). However, the recurrence rate of febrile UTI within 6 months was higher(25.8%) than control patients(6.6%). Conclusion: Epidemiologic study is required to find out any new risk factors of community-acquired ESBL(+) E. coli UTI and changes in selection of empirical antibiotics should be considered.

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The Effect of Acupuncture on the Intestinal Smooth Muscle Dysfunction Caused by Enteric Bacterial Infection in Weaning Piglet (돼지의 세균성 위장관 질환에 따른 평활근 기능 이상에 대한 침술의 효과)

  • Kim, Tae-wan;La, Jun-ho;Sung, Tae-sik;Kang, Jeong-woo;Nam, Tchi-chou;Choi, Min-cheol;Yoon, Yeo-sung;Yang, Il-suk
    • Korean Journal of Veterinary Research
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    • v.43 no.3
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    • pp.415-421
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    • 2003
  • It is now generally accepted that acupuncture is effective in diarrhea caused by bacterial infection. However, its effect on the intestinal smooth muscle dysfunction is not clear. Therefore, we investigated the effect of acupuncture therapy at Jiao-chao (GV-1) on the intestinal muscle dysfunction in weaning piglets orally infected by Escherichia coli. The animals are divided into four groups; 1) E. coli + no-treatment, 2) E. coli + antibiotic, 3) E. coli + acupuncture, 4) normal group. In the three E. coli infected groups, low frequency electrical field stimulation (EFS, 1 Hz) provoked triphasic responses composed of initial relaxation followed by on-contraction and off-contraction. While in the normal group, EFS (1Hz) induced biphasic responses composed of relaxation during the stimulation and off-contraction. At the high frequency (16Hz) EFS, both on-contraction and off-contraction of the E. coli + antibiotic, E. coli + acupuncture and the normal group were larger than those of the E. coli + no-treatment group. In the non-adrenergic non-cholinergic (NANC) condition, only biphasic responses occurred to EFS in all experimental groups and the off-contraction of E. coli + antibiotic, E. coli + acupuncture and the normal group were larger than those of the E. coli + no-treatment group. The response to carbachol of those three groups was also significantly greater than that of the E. coli + no-treatment group. These results suggest that acupuncture is as effective as antibiotic in the dysfunction of colonic circular muscle caused by E. coli infection. The maintenance of contractile neuromuscular transmission seems to be involved in the mechanism of the acupuncture effects on diarrhea.

Geochemical Analysis and Fates of Pathogenic Indicating Bacteria on Seawater Intrusion in a Sand Box Model (인공 대수층내에서 발생하는 해수침투의 지화학적 분석 및 병원성 지표 미생물의 사멸 특성)

  • Lee, So-Jung;Park, Hun-Ju;Sung, Eun-Hae;So, Myung-Ho;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.4
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    • pp.385-392
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    • 2008
  • In this study, seawater intrusion was assessed employing a kind of biological parameters such as Escherichia coli and Enterococcus faecalis while lab-prepared reclaimed water was recharged to prevent seawater intrusion. Chemical factors indicating seawater intrusion such as Cl$^-$, Ca$^{2+}$, Mg$^{2+}$ and specific conductivity were also simultaneously investigated where an ion exchange between a matrix in artificial aquifer and cations in solution was estimated. Both Escherichia coli and Enterococcus faecalis were shown to be very sensitive against degree of salinity during saline water intrusion. Enterococcus faecalis more strongly resisted against salinity than that of Escherichia coli. The ratio of Enterococcus faecalis divided by E. coli in the process of seawater intrusion increased up to more than 50$\sim$100 times in 18 hours whereas E. coli was died off more than 90% during pumping and recharge rate kept at 10 mL/min. However, when the rates of both recharge and pumping was kept at 5 mL/min, Enterococcus faecalis / Escherichia coli was sustained in the range of 2.5$\sim$5.0, while Escherichia coli showed dimished death rate. Chemical factors such as Cl$^-$, Ca$^{2+}$, Mg$^{2+}$ and specific conductivity showed more than 0.9 of high correlation each other well explaining the degree of seawater intrusion. The degree of ion exchange between artificial aquifer and saline water can be efficiently interpreted by both minus $\Delta$Na, $\Delta$Mg variation and positive $\Delta$Ca variation.

Prophylactic Effects of Bifidobacterium longum HY8001 against Escherichia coli O157:H7 and Salmonella typhimurium DT104 Enteric Infection and Evaluation of Vero Cytotxin Neutralizing Effects (Bifidobacterium longum HY8001 균주의 Escherichia coli O157:H7과 Salmonella typhimurium DT104 장곤 내 감염 예방효과 및 Vero cytotoxin 중화효과)

  • 양수진;윤장원;서근석;구혜정;김소현;배형석;백영진;박용호
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.419-425
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    • 1999
  • Prophylactic effects of Bifidobacterium longum HY8001, Korean isolate, against Escherichia coli O157:H7 and Salmonella typhimurium DT104 enteric infection were examined at four groups of specific pathogen free(SPF)-ICR mouse for each pathogen. B. longum HY8001+B. typhimurium DT104+B. longum HY8001(BL+ST+BL) group and B. longum HY8001+E. coli O157:H7+B. longum HY8001(BL+E+BL) group were fed with B. longum HY8001 before and after E. coli O157:H7 or s. typhimurium DT104 challenge, while B. longum HY8001+S. typhimurium DT104(BL+ST) and B. longum HY8001+e. coli O157:H7(BL+E) groups were fed with B. longum HY8001 only before E. coli O157:H7 or S. typhimurium DT104 challenge. E. coli O157:H7(E) and S. typhimurium DT104(ST) groups were challenged with each pathogen without B. longum HY8001 administration and control groups were administered with phosphate buffered solution(PBS). After the oral administration with B. longum HY8001(109cfu), th emice were challenged with E. coli O157:H7(2$\times$1010cfu) or S. typhimurium DT104(108cfu) and the mortality rate and the fecal shedding of challenged pathogen were also examined define the reactivity of the B. longum HY8001. Production of toxin neutralizing substance(s) of B. longum HY8001 was determined by cell cytotoxicity assay using Vero cells. Fecal shedding of th eS. typhimurium DT104 was significantly decreased in BL+ST+BL group fed with B. longum HY8--1 before and after challenge(p<0.05), while the fecal shedding s of S. typhimurium DT104 in BL+ST and St groups remained more than 106cfu. the protective effect of the B. longum HY8001 against E. coli O157:H7 was significantly high only in BL+E+BL group fed with b. longum Hy8001 before and after E. coli O157:H7 challenge from the result of fecal E. coli O157:H7 isolation rate, mortality rate, and intestinal contents culture to detect E. coli O157:H7. the mortality rate of the BL+e and E groups. The cytopathic effect (CPE) of the Vero cytotoxin (Shiga like toxin I & II) in Vero cell was neutralized in B. longum HY8001 culture supernatant added wells which indicate the presence of soluble Vero cytotxin neutralizing substance(s) in B. longum HY8001 culture suprnatant.

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Physiological Function of a DNA-Binding Protein from Starved Cells in Combating Diverse External Stresses in Escherichia coli (대장균 세포 내 다양한 외부 스트레스에 대한 DPS 단백질의 생리적 기능)

  • Lee, Joo Hyeong;Cheong, Su Jin;Oh, Hun Taek;Kim, Woe Yeon;Jung, Young Jun
    • Journal of Life Science
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    • v.23 no.4
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    • pp.479-486
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    • 2013
  • The DNA-binding protein from starved cells (DPS), originally identified as a DNA binding protein in Escherichia coli, is known to play an important role in DNA protection. The aim of this study was to evaluate the functional roles of DPS in E. coli against various kinds of external stresses by comparing the properties of wild-type E. coli cells and dps knockout mutant E. coli (${\Delta}dps$) cells. Under various stress conditions, we measured the cell growth of the wild-type E. coli and the dps knockout mutant E. coli (${\Delta}dps$) cells using a UV spectrophotometer. The growth rate of the cells was compared to investigate the functional roles of the DPS protein in E. coli. In comparison to the properties of the wild-type E. coli cells, the dps knockout mutant E. coli (${\Delta}dps$) cells showed highly sensitive phenotypes under various stress conditions, such as heat shock, acidic pH, nutrient deficiency, and different concentrations of reactive oxygen species (ROS), suggesting that DPS plays key roles in E. coli in combating diverse external stresses. The DPS DNA-binding protein in E. coli plays crucial roles in bacterial cell growth and in the protection of the cells from environmental stresses by tightly binding and preserving their DNA molecules.

Distributional Characteristics of Escherichia coli at Nakdong River Mouth and Busan Coastal Area (낙동강 하구와 부산연안해역에서 대장균의 해역별 분포특성)

  • Baek, Seung Ho;Lee, Min Ji;Yoon, Dongyoung
    • Journal of Marine Life Science
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    • v.1 no.1
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    • pp.1-7
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    • 2016
  • In order to assess seasonal and geographical characteristics of pollutant Escherichia coli, we investigated its distribution in Nakdong River mouth and Busan coastal water from February 2013 to November 2015. The coastal area was divided into five different zones (I-V) based on the pollutant level and geographical characteristics. During the study periods, water temperature and salinity varied from 7.50 to 27.64℃ and 16.82 to 34.82 psu, respectively. The annual water temperature variation was characterized in temperate zone. The salinity was significantly (p<0.05) decreased in zone IV and zone III after heavy rain during summer season in 2014, resulting led to elevated E. coli biomass. The highest colony formation of E. coli was recorded at 6,000 cfu l-1 during autumn at station 1 (zone I). On the other hands, during all seasons of 2015, E. coli abundances were kept to be low level in zone III. The E. coli was not significantly (p>0.05) correlated with water temperature. However, the salinity was significantly (r=-0.53, p<0.05) correlated with the E. coli, implying that salinity plays a crucial role in the proliferation of E. coli. Consequently, E. coli in western Busan coastal water might have been significantly promoted by pollutant sources from Nakdong Rive discharge during the spring and summer rainy seasons depending on annual rainfall variations. On the other hands, E. coli in station 1 (i.e., Suyeong Bay) was obviously high due to influences of discharge water from municipal wastewater treatment plant. However, there was no clear seasonality of E. coli.

Effect of Disinfection Process Combination on E. coli Deactivation and Oxidants Generation (E. coli 불활성화와 산화제 생성에 미치는 소독 공정 결합의 영향)

  • Kim, Dong-Seog;Park, Young-Seek
    • Journal of Environmental Science International
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    • v.20 no.7
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    • pp.891-898
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    • 2011
  • The aim of this research was to evaluate the effect of combination of disinfection process (electrolysis, UV process) on Escherichia coli (E. coli) disinfection and oxidants (OH radical, $ClO_2$, HOCl, $H_2O_2$ and $O_3$) generation. The effect of electrolyte type (NaCl, KCl and $Na_2SO_4$) on the E. coli disinfection and oxidants generation were evaluated. The experimental results showed that performance of E. coli disinfection of electrolysis and UV single process was similar. Combination of electrolysis and UV process enhanced the E. coli disinfection and 4-carboxybenzaldehyde (4-CBA, indicator of the generation of OH radical) degradation. It is clearly showed synergy effect on disinfection and OH radical formation. However chlorine ($ClO_2$, HOCl) and oxygen type ($H_2O_2$, $O_3$) oxidants were decreased with the combination of two process. In electrolysis + UV complex process, electro-generated $H_2O_2$ and $O_3$ were reacted with UV light of UV-C lamp and increased 4-CBA degradation(increase OH radical). Disinfection of electrolyte of chlorine type was higher than that of the sulfate type electrolyte due to the higher generation of OH radical and oxidants.

Escherichia coli-Derived Uracil Increases the Antibacterial Activity and Growth Rate of Lactobacillus plantarum

  • Ha, Eun-Mi
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.975-987
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    • 2016
  • Lactobacillus plantarum (L. plantarum) is a representative probiotic. In particular, L. plantarum is the first commensal bacterium to colonize the intestine of infants. For this reason, the initial settlement of L. plantarum can play an important role in determining an infant's health as well as their eventual health status as an adult. In addition, L. plantarum combats pathogenic infections (such as Escherichia coli (E. coli), one of the early pathogenic colonizers in an unhealthy infant gut) by secreting antimicrobial substances. The aim of this research was to determine how L. plantarum combats E. coli infection and why it is a representative probiotic in the intestine. Consequently, this research observed that E. coli releases uracil. L. plantarum specifically recognizes E. coli-derived uracil, which increases the growth rate and production of antimicrobial substance of L. plantarum. In addition, through the inhibitory activity test, this study postulates that the antimicrobial substance is a protein and can be considered a bacteriocin-like substance. Therefore, this research assumes that L. plantarum exerts its antibacterial ability by recognizing E. coli and increasing its growth rate as a result, and this phenomenon could be one of the reasons for L. plantarum settling in the intestine of infants as a beneficial bacterium.

Immunosensor for Detection of Escherichia coli O157:H7 Using Imaging Ellipsometry

  • Bae Young-Min;Park Kwang-Won;Oh Byung-Keun;Choi Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1169-1173
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    • 2006
  • Imaging ellipsometry (IE) for detection of binding of Escherichia coli O157:H7 (E. coli O157:H7) to an immunosensor is reported. A protein G layer, chemically bound to a self-assembled layer of 11-mercaptoundecanoic acid (11-MUA), was adopted for immobilization of monoclonal antibody against E. coli O157:H7 (Mab). The immobilization of antibody was investigated using surface plasmon resonance. To fabricate antibody spots on a gold surface, protein G solution was spotted onto the gold surface modified with an 11-MUA layer, followed by immobilizing Mab on the protein G spot. Ellipsometric images of the protein G spot, the Mab spot, and Mab spots with binding of E. coli O157:H7 in various concentrations were acquired using the IE system. The change of mean optical intensity of the Mab spots in the ellipsometric images indicated that the lowest detection limit was $10^3$CFU/ml for E. coli O157:H7. Thus, IE can be applied to an immunosensor for detection of E. coli O157:H7 as a detection method with the advantages of allowing label-free detection, high sensitivity, and operational simplicity.