• Title/Summary/Keyword: Pseudomonas Aeruginosa

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Sensitive and Enzyme-Free Pseudomonas aeruginosa Detection and Isolation via DNAzyme Cascade Triggered DNA Tweezer

  • Furong Liu;Jingyuan Xu;Lihua Yang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.9
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    • pp.1919-1925
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    • 2024
  • Effective isolation and sensitive detection of Pseudomonas aeruginosa (P. aeruginosa) is crucial for the early diagnosis and prognosis of various diseases, such as urinary tract infections. However, efficient isolation and simultaneous detection of P. aeruginosa remains a huge challenge. Herein, we depict a novel fluorescence assay for sensitive, enzyme-free detection of P. aeruginosa by integrating DNAzyme cascade-induced DNA tweezers and magnetic nanoparticles (MNPs)-based separation. The capture probe@MNPs is capable of accurately identifying target bacteria and transporting the bacteria signal to nucleic acid signals. Based on the DNAzyme cascade-induced DNA tweezers, the nucleic acid signals are extensively amplified, endowing the method with a high sensitivity and a low detection limit of 1 cfu/mL. In addition, the method also exhibits a wide detection of six orders of magnitudes. The proposed method could be extended to other bacteria detection by simply changing the aptamer sequence. Taking the merit of the high sensitivity, greatly minimized detection time (less than 1.5 h), enzyme-free characteristics, and stability, the proposed method could be potentially applied to diagnosing and preventing diseases caused by pathogenic bacteria.

Antimicrobial Activity of Pseudomonas aeruginosa BCNU 1204 and Its Active Compound (Pseudomonas aeruginosa BCNU 1204의 항균활성과 활성 물질)

  • Shin, Hwa Jin;Joo, Woo Hong
    • Journal of Life Science
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    • v.29 no.1
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    • pp.84-89
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    • 2019
  • Previous screening of novel antibacterial agents revealed that some bacterial isolates exhibited antibiotic activity against both gram-positive and gram-negative bacteria and that they showed antibacterial activity, even against methicillin-resistant Staphylococcus aureus (MRSA). Among these isolates, one bacterial strain, BCNU 1204, was identified as Pseudomonas aeruginosa using phenetic and phylogenetic analysis, based on 16S ribosomal RNA gene sequences. The maximum productivities of antimicrobial substances of BCNU 1204 were obtained after being cultured at $35^{\circ}C$ and pH 7.0 for 4 d in King's medium B (KMB). Dichloromethane (DCM) and ethylacetate (EA) extracts of P. aeruginosa BCNU 1204 exhibited strong antimicrobial activity, particularly against gram-positive bacteria. The EA extracts exhibited broad-spectrum activity against antibiotic resistant strains. Fraction 5-2, was obtained by recycling preparative liquid chromatography (LC) and preparative thin-layer chromatography (TLC) and was identified as phenazine-1-carboxylic acid belonging to phenazines using gas chromatography and mass spectrometry (GC/MS). Its minimum inhibitory concentration (MIC) values were $25{\mu}g/ml$, $50{\mu}g/ml$, ${\geq}25{\mu}g/ml$, and ${\geq}50{\mu}g/ml$ for MRSA CCARM 3089, 3090, 3091, and 3095 strains, respectively. P. aeruginosa BCNU 1204 may be a potential resource for the development of anti-MRSA antibiotics. Additional research is required to identify the active substance from P. aeruginosa BCNU 1204.

Purification and Properties of Biosurfactant from Pseudomonas aeruginosa KK-7 (Pseudomonas aeruginosa가 생산하는 biosurfactant의 분리 및 특성)

  • 김대원;김민주;강상모
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.337-345
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    • 1995
  • Biosurfactants produced by Pseudomonas aeroginosa KK-7 were purified and their properties were studied. The bacterial surfactant was seperated into two sorts of biosufactants (Type I, 11) by silica gel column chromatograpgy. On the basis of physiochemical analysis, Type I was found to be mixture of two glycolipids with M.W. 800, and Type II was peptide with M.W. 1300. The Type 11 biosurfactant was compose of glutamic acid, proline, glycine, leucine, histidine. The crude extract was used to dertermine some properties as a surfactant. The biosurfactant had the properties as stronger emulsification agent and a stronger stabilizing agent emulsion than any other surfactants tested.

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Growth Suppression of Microcystis aeruginosa by Pseudomonas aeruginosa AJ1 (Pseudomonas aeruginosa AJ1에 의한 Microcystis aeruginosa의 성장제어)

  • Kim, Sun-Jung;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.362-367
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    • 2009
  • Among total 176 strains with antialgal effects isolated from So-ok stream in Korea, Pseudomonas aeruginosa AJ1 showed the highest removal efficiency for an algal species Microcystis aeruginosa (clear zone of diameter 50.0 mm on algal lawn after 20 days). The algal growth was suppressed even when the supernatant of AJ1 culture was applied, suggesting that extracellular substances are responsible for its antialgal activity. The removal activity of AJ1 was optimal under the following condition: pH 8, $30^{\circ}C$, and mannitol as a carbon source. The antialgal activity of AJ1 appeared to be dependent of the growth phase of M. aeruginosa, i.e., the highest at the early phase, but not its own phase. As expected, the algicidal effect was improved as the amount of the treated supernatant was increased; the highest removal efficiency (80.3%) was achieved when 40 ml/L of the supernatant was used. Interestingly, however, the removal rate was opposite. The highest removal rate ($8.2{\mu}g$ chl-a/ml supernatant/day) was achieved when low concentration (10 ml/L) was applied. These results suggest that P. aeruginosa AJ1 is a promising biological agent to control the problematic algal bloom.

Patterns of Antimicrobial Resistance and Genotyping of Carbapenemase-producing Imipenem-nonsusceptible Pseudomonas aeruginosa (Imipenem 비감수성 Carbapenemase 생성 Pseudomonas aeruginosa에 의한 항생제 내성유형과 분자생물학적인 특성)

  • Lee, Jin-Hee;Lee, Gyusang;Lim, Kwanhun;Eom, Yong-Bin;Kim, Shin-Moo;Kim, Jong-Bae
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.2
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    • pp.71-80
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    • 2010
  • Pseudomonas aeruginosa are important nosocomial pathogens. Their resistance to carbapenem is increasing and causing concerns in Korea. An increasing prevalence of carbapenem resistance mediated by acquired carbapenemase is being reported. Over a 10 month-period from July 2007 to April 2008, 32 strains of imipenem-nonsusceptible P. auruginosa were isolated from Kangwon National University Hospital. To determine the prevalence and genotypes of the carbapenemase-producing clinical isolates, the antibiotic susceptibility was determined by Microscan Walkaway 96 SI System and the carbapenem activity was detected by the modified Hodge test and the imipenem-EDTA-SMA double-disk synergy test. The metallo-${\beta}$-lactamase gene and OXA-type ${\beta}$-lactamase gene reported in Korea were detected by PCR. As for the result of PCR, 30 isolates of P. aeruginosa were found to have $bla_{IMP-1}$-like and 1 isolate was found to have $bla_{IMP-1}$-like and $bla_{IMP-2}$. No clinical isolates were found to have $bla_{SIM-1}$, $bla_{OXA-23}$-like and $bla_{OXA-24}$-like. Random amplified polymorphic DNA (RAPD)-PCR and dendrogram for genetical similarity to band patterns of each clinical isolates were examined. P. aeruginosa were grouped into 7 clusters of up to 50% of similarity index. In the P. aeruginosa group, PS3 was resistant to the most antibiotics, PS1 was susceptible to the most antibiotics. PS7 was resistant to aztreonam unlike other groups. This is the first report of prevalence of carbapenemase in Chuncheon.

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Biodegradation Enhancement of The Mixture of Kerosene and Diesel by using Biosurfactant from Pseudomonas aeruginosa F722 (Pseudomonas aeruginosa F722부터 유래된 biosurfactant를 이용한 등.경유 혼합물의 생분해율 향상)

  • ;;;skubo Motoki
    • KSBB Journal
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    • v.18 no.6
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    • pp.529-535
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    • 2003
  • We studied degradation effects of hydrophobic substrate such as kerosene and diesel by adding a biosurfactant originated from Pseudomonas aeruginosa F722 and chemical surfactants (Tween 80 and detergent) with aeration. The surface tensions of the biosurfactant, Tween 80 and detergent were 30mN/m, 39mN/m and 31mN/m, respectively. When the concentration of biosurfactant added in C-medium was 0.01 and 0.15%(w/v), the ratios of hydrocarbon degradation were 94.3% and 94.2% respectively. It was 6.2%(w/v) higher than when the concentrations of added biosurfactant were 0.05, 0.1 and 0.2%. The degradation ratios of the chemical surfactants (Tween 80 and detergent) were 94.5% and 93.5% respectively. The effects of the biosurfactant and chemical surfactants were similar on the degradation ratio in mixtures of kerosene and diesel. However, the population of viable p. aeruginosa F722 at the end of the cultivation period was twice as higher in the biosurfactant than that in the chemical surfactant. We also studied the effect of aeration (0.5vvm) on the degradation ratio. The biosurfactant addition experiment was conducted with 0.5vvm air, 35$^{\circ}C$, 150rpm, pH 8.0, 3days, 1.0% (w/v) substrate. When p. aeruginosa F722 and 0.15%(w/v) biosurfactant were added, the degradation ratio of hydrocarbon was 94.8%. Without p. aeruginosa F722, it was 68%. Thus, with aeration, the degradation ratio of hydrocarbon was increased by 26.8%. In addition, the cultivation time was shortened by 1/3. The degradation ratios of hydrocarbon in shaking culture (cultivation time; 3days) and stationary culture (cultivation time; 10days) were 94.8 and 93.7% respectively. Thus, the addition of biosurfactant and aeration enhanced the degradation of hydrocarbon originated kerosene and diesel.

Antibacterial Effects and Cellular Responses of Imipenem-resistant Pseudomonas aeruginosa Exposed to Green Tea Polyphenols (녹차 폴리페놀에 노출된 Imipenem 내성 Pseudomonas aeruginosa의 항균효과 및 세포반응)

  • Song, You-Jin;Cho, Yun-Seok;Oh, Kye-Heon
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.198-206
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    • 2010
  • The aim of this work was to investigate the synergically bactericidal effects and cellular responses of tea polyphenols (TPP) and imipenem on imipenem-resistant Pseudomonas aeruginosa. Imipenem-resistant Ps. aeruginosa was isolated from patient in hospital. The bactericidal effects of TPP and imipenem were evaluated on the basis of its minimum inhibitory concentrations (MIC). The combined use of TPP and imipenem resulted in 16-fold and 8-fold reductions in the MICs of imipenem for the imipenem-susceptible and imipenem-resistant Ps. aeruginosa, respectively. The bactericidal effects of the imipenem and TPP against the Ps. aeruginosa was evaluated using the time-kill assay. The synergetic effects of the combinations of TPP and imipenem against Ps. aeruginosa were confirmed. Western blot using anti-DnaK and anti-GroEL monoclonal antibodies was performed to investigate the expression of stress shock proteins (SSPs) in imipenem-susceptible and imipenem-resistant strains exposed to TPP. The amount of SSPs were induced as the exposure time increased and decreased. The molecular weights of DnaK and GroEL were 70 kDa and 60 kDa, respectively. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides (LPS) increased or decreased in the strain treated to different concentrations and exposing periods of TPP. Scanning electron microscopic analysis demonstrated the presence of umblicated and wrinkled surfaces for cells treated with TPP or imipenem.

The Experimental Study on Anti-bacterial Potency of Sean-tang on Three species of causative bacteria of Keratitis (세안탕(洗眼湯)이 다종(多種)의 각막염(角膜炎) 유발균(誘發菌)에 미치는 효과(效果)에 관한 실험적 연구)

  • Kim, Yong-Kuk;Jung, Soon-Young;Yoo, Jin-Gon;Seo, Hyung-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.19 no.1
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    • pp.11-20
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    • 2006
  • Objective : This experimental study was performed to investigate the effect of herbal eye drops, Sean-tang and Coptidis rhizoma on Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa keratitis. Methods : After administering herbal eye drops (Sean-tang, Coptidis rhizoma) on Staphylococcus ausreus, Staphylococcus. epidermidis and Pseudomonas aeruginosa, I measured MIC and the size of inhibition zone. MIC was measured by dropping to $50{\mu}{\ell}$ according to density(l00%, 10%, 1%, 0.1%). Anti-bacterial potency was measured by the size of inhibition zone with change of volume. Result : On Staphylococcus aureus, Staphylococcus epidermidis, Coptidis rhizoma showed the highest anti-bacterial potency and on Pseudomonas aeruginosa, Sean-tang, Coptidis rhizoma was nor anti-bacterial potency. Conclusions : The present author think that the herbal eye drops, Sean-tang and Coptidis rhizoma can be used to cure Staphylococcus aureus, Staphyloccus epidermidis keratitis and if further study is performed, the use of the herbal eye drops will be valuable and beneficial in the clinical medicines.

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Combined action of Aminoglycoside and Cephalosporin Against Pseudomonas aeruginosa (녹농균에 대한 Aminoglycoside계와 Cephalosporin계의 병합작용)

  • Oh, Jong-Suk;Ahn, Tai-Hew
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.3
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    • pp.375-380
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    • 1986
  • Thirty-one strains of Pseudomonas aeruginosa were submitted to the synergistic activity test of amikacin(AK) and gentamicin(GM) combined with moxalactam(MX), ceftizoxime(CTZ) or cefoperazone(CFZ). The minimal inhibitory concentrations(MICs) of each drug and drugs combined in various ratios were measured by checkerboard dilution method. The synergism was determined through analysing the MIC distribution curve on isobologram and calculating the fractional inhibitory concentration index(FICI). MICs of GM, AK, MX, CFZ and CTZ against the 31 tested strains were distributed from $12.5{\mu}g/ml$ to $800{\mu}g/ml$, from $0.8{\mu}g/ml$ to $25{\mu}g/ml$, from $3.1{\mu}g/ml$ to $50{\mu}g/ml$, from $3.1{\mu}g/ml$ to $400{\mu}g/ml$, and from $12.5{\mu}g/ml$ to $100{\mu}g/ml$, respectively. The rate synergism of each drug combination by means of FICl was 45.5% in GM-MX, 36.4% in GM-CFZ, 63.6% in GM-CTZ, 48.6% in AK-MX, 35.3% in AK-CFZ, and 35.7% in AK-CTZ combination. Thus, it is suggested that Pseudomonas aeruginosa may effectively be inhibited by various aminoglycoside and cephalosporin combinations.

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