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Sample records for aeruginosa requiring removal

  1. Pseudomonas aeruginosa bacteriophage PA1Ø requires type IV pili for infection and shows broad bactericidal and biofilm removal activities.

    Science.gov (United States)

    Kim, Shukho; Rahman, Marzia; Seol, Sung Yong; Yoon, Sang Sun; Kim, Jungmin

    2012-09-01

    We isolated a new lytic Pseudomonas aeruginosa phage that requires type IV pili for infection. PA1Ø has a broad bactericidal spectrum, covering Gram-positive and Gram-negative bacteria, and can eradicate biofilm cells. PA1Ø may be developed as a therapeutic agent for biofilm-related mixed infections with P. aeruginosa and Staphylococcus aureus.

  2. Characterization of Glutamine-Requiring Mutants of Pseudomonas aeruginosa

    NARCIS (Netherlands)

    Janssen, Dick B.; Joosten, Han M.L.J.; Herst, Patricia M.; Drift, Chris van der

    1982-01-01

    Revertants were isolated from a glutamine-requiring mutant of Pseudomonas aeruginosa PAO. One strain showed thermosensitive glutamine requirement and formed thermolabile glutamine synthetase, suggesting the presence of a mutation in the structural gene for glutamine synthetase. The mutation conferri

  3. Removal of Microcystis aeruginosa using nano-Fe3O4 particles as a coagulant aid.

    Science.gov (United States)

    Zhang, Bo; Jiang, Dan; Guo, Xiaochen; He, Yiliang; Ong, Choon Nam; Xu, Yongpeng; Pal, Amrita

    2015-12-01

    Blue-green algae bloom is of great concern globally since they adversely affect the water ecosystem and also drinking water treatment processes. This work investigated the removal of Microcystis aeruginosa (M. aeruginosa) by combining the conventional coagulant polyaluminum chloride (PACl) with nano-Fe3O4 particles as a coagulant aid. The results showed that the addition of nano-Fe3O4 significantly improved the removal efficiency of M. aeruginosa by reducing the amount of PACl dosage and simultaneously hastening the sedimentation. At the M. aeruginosa density of an order of magnitude of 10(7), 10(6), and 10(5) pcs/mL, respectively, the corresponding PACl dose of 200, 20, and 2 mg/L and the mass ratio of PACl to nano-Fe3O4 of 4:1, the removal efficiency of M. aeruginosa could be increased by 33.0, 44.7, and 173.1%, respectively. Compared to PACl, PACl combined with the nano-Fe3O4 as a coagulant aid had higher removal efficiency at a wider pH range. SEM images showed that nano-Fe3O4 first combined with PACl to form clusters and further generated the flocs with algae. Results from the laser particle analyzer further suggested that the floc size increased with the addition of nano-Fe3O4. It was noted that the addition of nano-Fe3O4 led to aluminum species change after PACl hydrolyzed in the algae solution, from Ala to Alb and Alc subsequently. As a coagulant aid, the nano-Fe3O4, in conjunction with PACl, apparently provided nucleation sites for larger flocs to integrate with M. aeruginosa. In addition, increased floc density improved the removal of M. aeruginosa.

  4. Bisphenol A removal by a Pseudomonas aeruginosa immobilized on granular activated carbon and operating in a fluidized bed reactor

    Energy Technology Data Exchange (ETDEWEB)

    Mita, Luigi [National Laboratory on Endocrine Disruptors, National Institute of Biostructures and Biosystems (INBB), Via P. Castellino, 111, 80131 Naples (Italy); Institute of Genetic and Biophysics “ABT”, Via P. Castellino, 111, 80131 Naples Italy (Italy); Grumiro, Laura [National Laboratory on Endocrine Disruptors, National Institute of Biostructures and Biosystems (INBB), Via P. Castellino, 111, 80131 Naples (Italy); Rossi, Sergio [Institute of Genetic and Biophysics “ABT”, Via P. Castellino, 111, 80131 Naples Italy (Italy); Bianco, Carmen; Defez, Roberto [Institute of Biosciences and BioResources, Via P. Castellino, 111, 80131 Naples (Italy); Gallo, Pasquale [Dipartimento di Chimica, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via della Salute 2, 80055 Portici, Naples (Italy); Mita, Damiano Gustavo, E-mail: mita@igb.cnr.it [National Laboratory on Endocrine Disruptors, National Institute of Biostructures and Biosystems (INBB), Via P. Castellino, 111, 80131 Naples (Italy); Institute of Genetic and Biophysics “ABT”, Via P. Castellino, 111, 80131 Naples Italy (Italy); Diano, Nadia [National Laboratory on Endocrine Disruptors, National Institute of Biostructures and Biosystems (INBB), Via P. Castellino, 111, 80131 Naples (Italy); Department of Experimental Medicine, Second University of Naples, Via S.M. di Costantinopoli, 16, 80138 Naples Italy (Italy)

    2015-06-30

    Highlights: • A fluidized bed reactor, filled with a Pseudomonas aeruginosa immobilized on GAC, has been used for BPA removal. • BPA removal resulted from a biological activated carbon (BAC) process. • Equations describing the results have been indicated. • BPA removal was analyzed as a function of time and biofilm reuse. - Abstract: Serratia rubidiae, Pseudomonas aeruginosa and Escherichia coli K12 have been studied for their ability of Bisphenol A removal from aqueous systems and biofilm formation on activated granule carbon. Mathematical equations for biodegradation process have been elaborated and discussed. P. aeruginosa was found the best strain to be employed in the process of Bisphenol A removal. The yield in BPA removal of a P. aeruginosa biofilm grown on GAC and operating in a fluidized bed reactor has been evaluated. The results confirm the usefulness in using biological activated carbon (BAC process) to remove phenol compounds from aqueous systems.

  5. Bisphenol A removal by a Pseudomonas aeruginosa immobilized on granular activated carbon and operating in a fluidized bed reactor.

    Science.gov (United States)

    Mita, Luigi; Grumiro, Laura; Rossi, Sergio; Bianco, Carmen; Defez, Roberto; Gallo, Pasquale; Mita, Damiano Gustavo; Diano, Nadia

    2015-06-30

    Serratia rubidiae, Pseudomonas aeruginosa and Escherichia coli K12 have been studied for their ability of Bisphenol A removal from aqueous systems and biofilm formation on activated granule carbon. Mathematical equations for biodegradation process have been elaborated and discussed. P. aeruginosa was found the best strain to be employed in the process of Bisphenol A removal. The yield in BPA removal of a P. aeruginosa biofilm grown on GAC and operating in a fluidized bed reactor has been evaluated. The results confirm the usefulness in using biological activated carbon (BAC process) to remove phenol compounds from aqueous systems.

  6. Complete Genome Sequence of Pseudomonas aeruginosa Podophage MPK7, Which Requires Type IV Pili for Infection.

    Science.gov (United States)

    Bae, Hee-Won; Cho, You-Hee

    2013-10-10

    We report the complete genome sequence of Pseudomonas aeruginosa podophage MPK7. It displays synteny to the P. aeruginosa phages of the Phikmvlikevirus genus, which includes phiKMV and LKA1. MPK7 requires type IV pili (TFP) for infection, suggesting the role of functional TFP as the receptor for this phage genus.

  7. The cytotoxin of Pseudomonas aeruginosa : Cytotoxicity requires proteolytic activation

    NARCIS (Netherlands)

    Orlik-Eisel, Gabriele; Lutz, Frieder; Henschen, Agnes; Eisel, Ulrich; Struckmeier, Martin; Kräuter, Josef; Niemann, Heiner

    1990-01-01

    The primary structure of a cytotoxin from Pseudomonas aeruginosa was determined by sequencing of the structural gene. The cytotoxin (31,700 Mr) lacks an N-terminal signal sequence for bacterial secretion but contains a pentapeptide consensus sequence commonly found in prokaryotic proteins which func

  8. Removal of Cr(VI from aqueous solution using Bacillus subtilis, Pseudomonas aeruginosa and Enterobacter cloacae

    Directory of Open Access Journals (Sweden)

    P. Sethuraman,

    2010-06-01

    Full Text Available The objective of this study is to investigate the removal efficiency of Cr(VI by Bacillus subtilis, Pseudomonas aeruginosa and Enterobacter cloacae from aqueous solution under different process conditions. Batch mode experiments were carried out as a function of solution pH, biosorbent dosage, Cr(VI concentration and contact time.The FT-IR spectra and SEM analysis of the biosorbent were recorded to analyse the number and position of the functional groups available for the binding of Cr(VI ions and to study the morphology of biosorbent. The batch isothermal equilibrium data were analyzed with Freundlich and Langmuir isotherm models. The kinetic models were examined with pseudo first order and pseudo second order kinetics. The results revealed that the Cr(VI is considerably adsorbed on bacterial biomass and it could be an economical method for the removal of Cr(VI from aqueous solution.

  9. Microcystin-LR removal from Microcystis aeruginosa using in natura sugarcane bagasse and activated carbon

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    Aline Rafaela de Almeida

    2016-03-01

    Full Text Available Microcystin-LR is a type of toxin released by the Microcystis aeruginosa cyanobacteria found in water sources used for human consumption. It can cause illness and even death if not completely removed in conventional water treatment. The retention of this toxin is often accomplished by the adsorption process in activated carbon in water treatment plants. In this study, a comparison was made between the retention of microcystin-LR by activated carbon and by sugarcane bagasse in natura applied as a bio-adsorbent. Adsorption experiments were performed after the physical and chemical properties of the bio adsorbent and the activated carbon were characterized. The adsorption performance was evaluated by the toxin removal efficiency and the maximum adsorption capacity. Average removal efficiencies of the toxin resulted in 65.25; 41.74 and 11.75% for the activated carbon and 24.15; 18.92 and 12.27% for the sugarcane bagasse for concentrations of 2.36, 3.33 and 3.83 µg L-1, respectively. The bio adsorbent presented removal efficiency for the toxin similar to that observed in the activated carbon for the concentration of 3.83 µg L-1. Maximum adsorption capacity obtained with better linear adjustment to the Freundlich isotherm was 6,047.84 µg g-1 (toxin concentration of 3.83 µg L-1 for sugarcane bagasse and 338.61 µg g-1 (toxin concentration of 2.36 µg L-1 for activated carbon.

  10. Isolation and characterization of Pseudomonas aeruginosa mutants requiring salicylic acid for pyochelin biosynthesis.

    OpenAIRE

    Ankenbauer, R G; Cox, C D

    1988-01-01

    Pseudomonas aeruginosa mutants requiring salicylic acid for pyochelin biosynthesis were isolated after chemical mutagenesis by plating on a siderophore detection medium. Like the wild type, these mutants incorporated 7-[14C]salicylic acid into pyochelin, demonstrating that salicylic acid is an intermediate in the biosynthesis pathway of pyochelin.

  11. Removal of toxic Co-EDTA complex by a halophilic solar-salt-pan isolate Pseudomonas aeruginosa SPB-1.

    Science.gov (United States)

    Paraneeiswaran, A; Shukla, Sudhir K; Subba Rao, T; Prashanth, K

    2014-01-01

    In this study, a promising bioremediation approach was developed to remove [Co(III)-EDTA](-) complex that is generated during the waste management process. Though several studies have been reported on bioremediation of cobalt, the removal of [Co(III)-EDTA](-) complex has not been tested. A [Co(III)-EDTA](-) resistant bacterium, Pseudomonas aeruginosa SPB-1 was isolated from the solar-salt-pan and physical parameters were optimized for its growth. The various studies showed that the removal of [Co(III)-EDTA](-) from the bulk liquid was due to the adsorption of the complex by the biomass. Using absorption/desorption isotherm over a range of pH (1-8), the maximum adsorption of [Co(III)-EDTA](-) was found to be at pH 7.0 and maximum desorption from the biomass occurred at pH 1.0, thus rendering an ion exchange property to P. aeruginosa SPB-1 biomass. P. aeruginosa SPB-1 biomass could be used as bio-resin that showed 80.4±3.27% adsorption capacity up to fourth cycle and the biomass was viable till the ninth cycle with 10.5±7.3% adsorption. Radiation tolerance potential i.e. D10 value for the strain was found to be ~300 Gy, which suggests the potential use of the bacterium in bioremediation of moderately active nuclear waste.

  12. Revealing the characteristics of a novel bioflocculant and its flocculation performance in Microcystis aeruginosa removal

    Science.gov (United States)

    Sun, Pengfei; Hui, Cai; Bai, Naling; Yang, Shengmao; Wan, Li; Zhang, Qichun; Zhao, Yuhua

    2015-12-01

    In the present work, a novel bioflocculant, EPS-1, was prepared and used to flocculate the kaolin suspension and Microcystis aeruginosa. We focused on the characteristics and flocculation performance of EPS-1, especially with regard to its protein components. An important attribute of EPS-1 was its protein content, with 18 protein types identified that occupied a total content of 31.70% in the EPS-1. Moreover, the flocculating activity of these protein components was estimated to be no less than 33.93%. Additionally, polysaccharides that occupied 57.12% of the total EPS-1 content consisted of four monosaccharides: maltose, D-xylose, mannose, and D-fructose. In addition, carbonyl, amino, and hydroxyl groups were identified as the main functional groups. Three main elements, namely C1s, N1s, and O1s, were present in EPS-1 with relative atomic percentages of 62.63%, 24.91%, and 10.5%, respectively. Zeta potential analysis indicated that charge neutralization contributed to kaolin flocculation, but was not involved in M. aeruginosa flocculation. The flocculation conditions of EPS-1 were optimized, and the maximum flocculating efficiencies were 93.34% within 2 min for kaolin suspension and 87.98% within 10 min for M. aeruginosa. These results suggest that EPS-1 could be an alternative to chemical flocculants for treating wastewaters and cyanobacterium-polluted freshwater.

  13. Potential application of aerobic denitrifying bacterium Pseudomonas aeruginosa PCN-2 in nitrogen oxides (NOx) removal from flue gas.

    Science.gov (United States)

    Zheng, Maosheng; Li, Can; Liu, Shufeng; Gui, Mengyao; Ni, Jinren

    2016-11-15

    Conventional biological removal of nitrogen oxides (NOx) from flue gas has been severely restricted by the presence of oxygen. This paper presents an efficient alternative for NOx removal at varying oxygen levels using the newly isolated bacterial strain Pseudomonas aeruginosa PCN-2 which was capable of aerobic and anoxic denitrification. Interestingly, nitric oxide (NO), as the obligatory intermediate, was negligibly accumulated during nitrate and nitrite reduction. Moreover, normal nitrate reduction with decreasing NO accumulation was realized under O2 concentration ranging from 0 to 100%. Reverse transcription and real-time quantitative polymerase chain reaction (RT-qPCR) analysis revealed that high efficient NO removal was attributed to the coordinate regulation of gene expressions including napA (for periplasmic nitrate reductase), nirS (for cytochrome cd1 nitrite reductase) and cnorB (for NO reductase). Further batch experiments demonstrated the immobilized strain PCN-2 possessed high capability of removing NO and nitrogen dioxide (NO2) at O2 concentration of 0-10%. A biotrickling filter established with present strain achieved high NOx removal efficiencies of 91.94-96.74% at inlet NO concentration of 100-500ppm and O2 concentration of 0-10%, which implied promising potential applications in purifying NOx contaminated flue gas.

  14. Characterization of Microcystis Aeruginosa immobilized in complex of PVA and sodium alginate and its application on phosphorous removal in wastewater

    Institute of Scientific and Technical Information of China (English)

    李飞; 毛文娟; 李雪; 王晓钰; 肖智华; 周耀渝; 曾光明

    2015-01-01

    Based on ecological niche theory, Microcystis Aeruginosa (MA) immobilized in the complex of polyvinyl alcohol (PVA) and sodium alginate (SA) crosslinked by CaCl2, was treated as a new kind of special species, and its properties were investigated. Chlorophyll a was used to characterize the bioactivity of the immobilized MA. Results reveal that the gel beads have mechanical strength and chemical stability even under non-sterile harsh conditions, which may be attributed to the rarely seen structure (including three different layers: dense surface, tubular-shaped divergent structure and honeycomb crystal lattice layer) of the immobilized MA determined by scanning electron microscope (SEM). SEM also displays that more quantity of MA is attached to the inwall after cultivation, which demonstrates that the MA within beads maintains high bioactivity. Removal capacities on phosphorous (P) removal in wastewater in the presence and absence of the BG-11 medium were examined, and the removal ratios are 80.3%and 76.7%, respectively, which indicates that the beads without providing ample nutrients still have high capacity of P removal. In addition, control experiment, utilizing polyvinyl alcohol and sodium alginate (PVA-SA) beads without immobilized MA, demonstrates that MA within beads plays the key role in absorbing P.

  15. Kirschner Wire Breakage during Removal Requiring Retrieval

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    Kai Yuen Wong

    2016-01-01

    Full Text Available Kirschner wires (K-wires are widely used for fixation of fractures and dislocations in the hand as they are readily available, reliable, and cost-effective. Complication rates of up to 18% have been reported. However, K-wire breakage during removal is rare. We present one such case illustrating a simple technique for retrieval. A 35-year-old male presented with a distal phalanx fracture of his right middle finger. This open fracture was treated with K-wire fixation. Postoperatively, he developed a pin site infection with associated finger swelling. The K-wire broke during removal with the proximal piece completely retained in his middle phalanx. To minimise risk of osteomyelitis, the K-wire was removed with a novel surgical technique. He had full return of hand function. Intraoperative K-wire breakage has a reported rate of 0.1%. In our case, there was no obvious cause of breakage and the patient denied postoperative trauma. On the other hand, pin site infections are much more common with reported rates of up to 7% in the hand or wrist. K-wire fixation is a simple method for bony stabilisation but can be a demanding procedure with complications often overlooked. It is important to be aware of the potential sequelae.

  16. Pseudomonas aeruginosa β-lactamase induction requires two permeases, AmpG and AmpP

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    Schneper Lisa

    2010-12-01

    Full Text Available Abstract Background In Enterobacteriaceae, β-lactam antibiotic resistance involves murein recycling intermediates. Murein recycling is a complex process with discrete steps taking place in the periplasm and the cytoplasm. The AmpG permease is critical to this process as it transports N-acetylglucosamine anhydrous N-acetylmuramyl peptides across the inner membrane. In Pseudomonadaceae, this intrinsic mechanism remains to be elucidated. Since the mechanism involves two cellular compartments, the characterization of transporters is crucial to establish the link. Results Pseudomonas aeruginosa PAO1 has two ampG paralogs, PA4218 (ampP and PA4393 (ampG. Topology analysis using β-galactosidase and alkaline phosphatase fusions indicates ampP and ampG encode proteins which possess 10 and 14 transmembrane helices, respectively, that could potentially transport substrates. Both ampP and ampG are required for maximum expression of β-lactamase, but complementation and kinetic experiments suggest they act independently to play different roles. Mutation of ampG affects resistance to a subset of β-lactam antibiotics. Low-levels of β-lactamase induction occur independently of either ampP or ampG. Both ampG and ampP are the second members of two independent two-gene operons. Analysis of the ampG and ampP operon expression using β-galactosidase transcriptional fusions showed that in PAO1, ampG operon expression is β-lactam and ampR-independent, while ampP operon expression is β-lactam and ampR-dependent. β-lactam-dependent expression of the ampP operon and independent expression of the ampG operon is also dependent upon ampP. Conclusions In P. aeruginosa, β-lactamase induction occurs in at least three ways, induction at low β-lactam concentrations by an as yet uncharacterized pathway, at intermediate concentrations by an ampP and ampG dependent pathway, and at high concentrations where although both ampP and ampG play a role, ampG may be of greater

  17. Prevalence of Multidrug Efflux Pump Requiring Ciprofloxacin, Ofloxacin and Pefloxacin as Substrates, Among Clinical Isolates of Pseudomonas aeruginosa

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    Omoregie, R.

    2007-01-01

    Full Text Available Forty two consecutive clinical isolates of Pseudomonas aeruginosa were screened for the presence of reserpine inhibited multidrug (MDR efflux pump, utilizing ciprofloxacin ofloxacin and/or pefloxacin as substrates, by determining the minimum inhibitory concentration in the presence and absence of 100 mg/L reserpine. The result showed that 50% of the Pseudomonas aeruginosa isolates possessed reserpine inhibited MDR efflux pump. MDR efflux pump requiring ofloxacin (40.48% were significantly (p<0.01 more among the isolates when compared with those requiring ciprofloxacin (16.67% or pefloxacin (11.90%. Only one isolate possessed reserpine inhibited MDR efflux pumps that utilize all three fluiroquinolones. Research into suitable combination of antibacterials and appropriate pump mactivators or antibacterials that are less likely to be substrate for MDR pumps is advocated.

  18. Dimeric c-di-GMP is required for post-translational regulation of alginate production in Pseudomonas aeruginosa.

    Science.gov (United States)

    Whitney, John C; Whitfield, Gregory B; Marmont, Lindsey S; Yip, Patrick; Neculai, A Mirela; Lobsanov, Yuri D; Robinson, Howard; Ohman, Dennis E; Howell, P Lynne

    2015-05-15

    Pseudomonas aeruginosa is an opportunistic human pathogen that secretes the exopolysaccharide alginate during infection of the respiratory tract of individuals afflicted with cystic fibrosis and chronic obstructive pulmonary disease. Among the proteins required for alginate production, Alg44 has been identified as an inner membrane protein whose bis-(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP) binding activity post-translationally regulates alginate secretion. In this study, we report the 1.8 Å crystal structure of the cytoplasmic region of Alg44 in complex with dimeric self-intercalated c-di-GMP and characterize its dinucleotide-binding site using mutational analysis. The structure shows that the c-di-GMP binding region of Alg44 adopts a PilZ domain fold with a dimerization mode not previously observed for this family of proteins. Calorimetric binding analysis of residues in the c-di-GMP binding site demonstrate that mutation of Arg-17 and Arg-95 alters the binding stoichiometry between c-di-GMP and Alg44 from 2:1 to 1:1. Introduction of these mutant alleles on the P. aeruginosa chromosome show that the residues required for binding of dimeric c-di-GMP in vitro are also required for efficient alginate production in vivo. These results suggest that the dimeric form of c-di-GMP represents the biologically active signaling molecule needed for the secretion of an important virulence factor produced by P. aeruginosa.

  19. The Pseudomonas aeruginosa Type III Translocon Is Required for Biofilm Formation at the Epithelial Barrier

    DEFF Research Database (Denmark)

    Tran, Cindy S; Rangel, Stephanie M; Almblad, Henrik;

    2014-01-01

    Clinical infections by Pseudomonas aeruginosa, a deadly Gram-negative, opportunistic pathogen of immunocompromised hosts, often involve the formation of antibiotic-resistant biofilms. Although biofilm formation has been extensively studied in vitro on glass or plastic surfaces, much less is known...... about biofilm formation at the epithelial barrier. We have previously shown that when added to the apical surface of polarized epithelial cells, P. aeruginosa rapidly forms cell-associated aggregates within 60 minutes of infection. By confocal microscopy we now show that cell-associated aggregates...... a previously unappreciated function for the type III translocon in the formation of P. aeruginosa biofilms at the epithelial barrier and demonstrate that biofilms may form at early time points of infection....

  20. Anaerobic survival of Pseudomonas aeruginosa by pyruvate fermentation requires an Usp-type stress protein

    DEFF Research Database (Denmark)

    Schreiber, K; Boes, N; Escbach, M;

    2006-01-01

    Recently, we identified a pyruvate fermentation pathway in Pseudomonas aeruginosa sustaining anaerobic survival in the absence of alternative anaerobic respiratory and fermentative energy generation systems (M. Eschbach, K. Schreiber, K. Trunk, J. Buer, D. Jahn, and M. Schobert, J. Bacteriol. 186......:4596-4604, 2004). Anaerobic long-term survival of P. aeruginosa might be essential for survival in deeper layers of a biofilm and the persistent infection of anaerobic mucus plaques in the cystic fibrosis lung. Proteome analysis of P. aeruginosa cells during a 7-day period of pyruvate fermentation revealed...... the induced synthesis of three enzymes involved in arginine fermentation, ArcA, ArcB, and ArcC, and the outer membrane protein OprL. Moreover, formation of two proteins of unknown function, PA3309 and PA4352, increased by factors of 72- and 22-fold, respectively. Both belong to the group of universal stress...

  1. Pseudomonas aeruginosa serA Gene Is Required for Bacterial Translocation through Caco-2 Cell Monolayers

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    Yasuda, Masashi; Nagata, Syouya; Yamane, Satoshi; Kunikata, Chinami; Kida, Yutaka; Kuwano, Koichi; Suezawa, Chigusa; Okuda, Jun

    2017-01-01

    To specify critical factors responsible for Pseudomonas aeruginosa penetration through the Caco-2 cell epithelial barrier, we analyzed transposon insertion mutants that demonstrated a dramatic reduction in penetration activity relative to P. aeruginosa PAO1 strain. From these strains, mutations could be grouped into five classes, specifically flagellin-associated genes, pili-associated genes, heat-shock protein genes, genes related to the glycolytic pathway, and biosynthesis-related genes. Of these mutants, we here focused on the serA mutant, as the association between this gene and penetration activity is yet unknown. Inactivation of the serA gene caused significant repression of bacterial penetration through Caco-2 cell monolayers with decreased swimming and swarming motilities, bacterial adherence, and fly mortality rate, as well as repression of ExoS secretion; however, twitching motility was not affected. Furthermore, L-serine, which is known to inhibit the D-3-phosphoglycerate dehydrogenase activity of the SerA protein, caused significant reductions in penetration through Caco-2 cell monolayers, swarming and swimming motilities, bacterial adherence to Caco-2 cells, and virulence in flies in the wild-type P. aeruginosa PAO1 strain. Together, these results suggest that serA is associated with bacterial motility and adherence, which are mediated by flagella that play a key role in the penetration of P. aeruginosa through Caco-2 cell monolayers. Oral administration of L-serine to compromised hosts might have the potential to interfere with bacterial translocation and prevent septicemia caused by P. aeruginosa through inhibition of serA function. PMID:28046014

  2. Video requirements plan for the HMT equipment removal system

    Energy Technology Data Exchange (ETDEWEB)

    Vargo, G.F. Jr.

    1995-02-01

    This document is the plan defining the video coverage requirements for the equipment removal event of the Hydrogen Mitigation Test (HMT) mixer pump currently installed in high level nuclear waste storage Tank 241-SY-101. When the mixer pump fails the removal and installation of a spare pump will be a time critical event. Since the success of the HMT mixer pump has resolved the DOE safety issue it is absolutely essential that mixing be restored to the tank in a short as time possible. Therefore, the removal of the failed pump and the installation of the spare pump must be anticipated and planned well in advance. The removal, containment, transporting, and storage of the failed pump is a very complex and hazardous task. The successful completion of this task will require careful planning and monitoring. Certain events, during the removal and subsequent installation of the new pump, will require video observation and storage for safety, documenting, training, and promotional use. Furthermore, certain events will require close monitoring and observation by the event directors and key supervisory personnel for the execution of specific tasks during the equipment removal event.

  3. Genes required for and effects of alginate overproduction induced by growth of Pseudomonas aeruginosa on Pseudomonas isolation agar supplemented with ammonium metavanadate.

    Science.gov (United States)

    Damron, F Heath; Barbier, Mariette; McKenney, Elizabeth S; Schurr, Michael J; Goldberg, Joanna B

    2013-09-01

    Pseudomonas aeruginosa is an opportunistic pathogen that can adapt to changing environments and can secrete an exopolysaccharide known as alginate as a protection response, resulting in a colony morphology and phenotype referred to as mucoid. However, how P. aeruginosa senses its environment and activates alginate overproduction is not fully understood. Previously, we showed that Pseudomonas isolation agar supplemented with ammonium metavanadate (PIAAMV) induces P. aeruginosa to overproduce alginate. Vanadate is a phosphate mimic and causes protein misfolding by disruption of disulfide bonds. Here we used PIAAMV to characterize the pathways involved in inducible alginate production and tested the global effects of P. aeruginosa growth on PIAAMV by a mutant library screen, by transcriptomics, and in a murine acute virulence model. The PA14 nonredundant mutant library was screened on PIAAMV to identify new genes that are required for the inducible alginate stress response. A functionally diverse set of genes encoding products involved in cell envelope biogenesis, peptidoglycan remodeling, uptake of phosphate and iron, phenazine biosynthesis, and other processes were identified as positive regulators of the mucoid phenotype on PIAAMV. Transcriptome analysis of P. aeruginosa cultures growing in the presence of vanadate showed differential expression of genes involved in virulence, envelope biogenesis, and cell stress pathways. In this study, it was observed that growth on PIAAMV attenuates P. aeruginosa in a mouse pneumonia model. Induction of alginate overproduction occurs as a stress response to protect P. aeruginosa, but it may be possible to modulate and inhibit these pathways based on the new genes identified in this study.

  4. Requirement of the Pseudomonas aeruginosa CbrA sensor kinase for full virulence in a murine acute lung infection model.

    Science.gov (United States)

    Yeung, Amy T Y; Janot, Laure; Pena, Olga M; Neidig, Anke; Kukavica-Ibrulj, Irena; Hilchie, Ashley; Levesque, Roger C; Overhage, Joerg; Hancock, Robert E W

    2014-03-01

    Pseudomonas aeruginosa is an opportunistic pathogen that is a major cause of respiratory tract and other nosocomial infections. The sensor kinase CbrA is a central regulator of carbon and nitrogen metabolism and in vitro also regulates virulence-related processes in P. aeruginosa. Here, we investigated the role of CbrA in two murine models of infection. In both peritoneal infections in leukopenic mice and lung infection models, the cbrA mutant was less virulent since substantially larger numbers of cbrA mutant bacteria were required to cause the same level of infection as wild-type or complemented bacteria. In contrast, in the chronic rat lung model the cbrA mutant grew and persisted as well as the wild type, indicating that the decrease of in vivo virulence of the cbrA mutant did not result from growth deficiencies on particular carbon substrates observed in vitro. In addition, a mutant in the cognate response regulator CbrB showed no defect in virulence in the peritoneal infection model, ruling out the involvement of certain alterations of virulence properties in the cbrA mutant including defective swarming motility, increased biofilm formation, and cytotoxicity, since these alterations are controlled through CbrB. Further investigations indicated that the mutant was more susceptible to uptake by phagocytes in vitro, resulting in greater overall bacterial killing. Consistent with the virulence defect, it took a smaller number of Dictyostelium discoideum amoebae to kill the cbrA mutant than to kill the wild type. Transcriptional analysis of the cbrA mutant during D. discoideum infection led to the conclusion that CbrA played an important role in the iron metabolism, protection of P. aeruginosa against oxidative stress, and the regulation of certain virulence factors.

  5. Requirement of the Pseudomonas aeruginosa CbrA Sensor Kinase for Full Virulence in a Murine Acute Lung Infection Model

    Science.gov (United States)

    Yeung, Amy T. Y.; Janot, Laure; Pena, Olga M.; Neidig, Anke; Kukavica-Ibrulj, Irena; Hilchie, Ashley; Levesque, Roger C.; Overhage, Joerg

    2014-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen that is a major cause of respiratory tract and other nosocomial infections. The sensor kinase CbrA is a central regulator of carbon and nitrogen metabolism and in vitro also regulates virulence-related processes in P. aeruginosa. Here, we investigated the role of CbrA in two murine models of infection. In both peritoneal infections in leukopenic mice and lung infection models, the cbrA mutant was less virulent since substantially larger numbers of cbrA mutant bacteria were required to cause the same level of infection as wild-type or complemented bacteria. In contrast, in the chronic rat lung model the cbrA mutant grew and persisted as well as the wild type, indicating that the decrease of in vivo virulence of the cbrA mutant did not result from growth deficiencies on particular carbon substrates observed in vitro. In addition, a mutant in the cognate response regulator CbrB showed no defect in virulence in the peritoneal infection model, ruling out the involvement of certain alterations of virulence properties in the cbrA mutant including defective swarming motility, increased biofilm formation, and cytotoxicity, since these alterations are controlled through CbrB. Further investigations indicated that the mutant was more susceptible to uptake by phagocytes in vitro, resulting in greater overall bacterial killing. Consistent with the virulence defect, it took a smaller number of Dictyostelium discoideum amoebae to kill the cbrA mutant than to kill the wild type. Transcriptional analysis of the cbrA mutant during D. discoideum infection led to the conclusion that CbrA played an important role in the iron metabolism, protection of P. aeruginosa against oxidative stress, and the regulation of certain virulence factors. PMID:24379284

  6. Use of RSM modeling for optimizing decolorization of simulated textile wastewater by Pseudomonas aeruginosa strain ZM130 capable of simultaneous removal of reactive dyes and hexavalent chromium.

    Science.gov (United States)

    Maqbool, Zahid; Hussain, Sabir; Ahmad, Tanvir; Nadeem, Habibullah; Imran, Muhammad; Khalid, Azeem; Abid, Muhammad; Martin-Laurent, Fabrice

    2016-06-01

    Remediation of colored wastewater loaded with dyes and metal ions is a matter of interest nowadays. In this study, 220 bacteria isolated from textile wastewater were tested for their potential to decolorize each of the four reactive dyes (reactive red-120, reactive black-5, reactive yellow-2, and reactive orange-16) in the presence of a mixture of four different heavy metals (Cr, Zn, Pb, Cd) commonly found in textile effluents. Among the tested bacteria, the isolate ZM130 was found to be the most efficient in decolorizing reactive dyes in the presence of the mixture of heavy metals and was identified as Pseudomonas aeruginosa strain ZM130 by 16S rRNA gene analysis. The strain ZM130 was highly effective in simultaneously removing hexavalent chromium (25 mg L(-1)) and the azo dyes (100 mg L(-1)) from the simulated wastewater even in the presence of other three heavy metals (Zn, Pb, Cd). Simultaneous removal of chromium and azo dyes ranged as 76.6-98.7 % and 51.9-91.1 %, respectively, after 180 h incubation. On the basis of quadratic polynomial equation and response surfaces given by the response surface methodology (RSM), optimal salt content, pH, carbon co-substrate content, and level of multi-metal mixtures for decolorization of reactive red-120 in a simulated textile wastewater by the strain ZM130 were predicted to be 19.8, 7.8, and 6.33 g L(-1) and a multi-metal mixture (Cr 13.10 mg L(-1), Pb 26.21 mg L(-1), Cd 13.10 mg L(-1), Zn 26.21 mg L(-1)), respectively. Moreover, the strain ZM130 also exhibited laccase and nicotinamide adenine dinucleotide (reduced)-dichlorophenolindophenol reductase (NADH-DCIP reductase) activity during the decolorization of reactive red-120. However, the laccase activity was found to be maximum in the presence of 300 mg L(-1) of the dye as compared to other concentrations. Hence, the isolation of this strain might serve as a potential bio-resource required for developing the strategies aiming at bioremediation of the

  7. Required ozone doses for removing pharmaceuticals from wastewater effluents

    DEFF Research Database (Denmark)

    Antoniou, Maria; Hey, Gerly; Rodríguez Vega, Sergio

    2013-01-01

    The aim of the this study was to investigate the ozone dosage required to remove active pharmaceutical ingredients (APIs) from biologically treated wastewater of varying quality, originated from different raw wastewater and wastewater treatment processes.Secondary effluents from six Swedish...... wastewater treatment plants (WWTP) were spiked with 42 APIs (nominal concentration 1μg/L) and treated with different O3 doses (0.5–12.0mg/L ozone) in bench-scale experiments.In order to compare the sensitivity of APIs in each matrix, the specific dose of ozone required to achieve reduction by one decade...... of each investigated API (DDO3) was determined for each effluent by fitting a first order equation to the remaining concentration of API at each applied ozone dose. Ozone dose requirements were found to vary significantly between effluents depending on their matrix characteristics.The specific ozone dose...

  8. Cyclic di-GMP-mediated repression of swarming motility by Pseudomonas aeruginosa PA14 requires the MotAB stator.

    Science.gov (United States)

    Kuchma, S L; Delalez, N J; Filkins, L M; Snavely, E A; Armitage, J P; O'Toole, G A

    2015-02-01

    The second messenger cyclic diguanylate (c-di-GMP) plays a critical role in the regulation of motility. In Pseudomonas aeruginosa PA14, c-di-GMP inversely controls biofilm formation and surface swarming motility, with high levels of this dinucleotide signal stimulating biofilm formation and repressing swarming. P. aeruginosa encodes two stator complexes, MotAB and MotCD, that participate in the function of its single polar flagellum. Here we show that the repression of swarming motility requires a functional MotAB stator complex. Mutating the motAB genes restores swarming motility to a strain with artificially elevated levels of c-di-GMP as well as stimulates swarming in the wild-type strain, while overexpression of MotA from a plasmid represses swarming motility. Using point mutations in MotA and the FliG rotor protein of the motor supports the conclusion that MotA-FliG interactions are critical for c-di-GMP-mediated swarming inhibition. Finally, we show that high c-di-GMP levels affect the localization of a green fluorescent protein (GFP)-MotD fusion, indicating a mechanism whereby this second messenger has an impact on MotCD function. We propose that when c-di-GMP level is high, the MotAB stator can displace MotCD from the motor, thereby affecting motor function. Our data suggest a newly identified means of c-di-GMP-mediated control of surface motility, perhaps conserved among Pseudomonas, Xanthomonas, and other organisms that encode two stator systems.

  9. Influence of rhamnolipids, produced by Pseudomonas aeruginosa NCAIM(P, B001380 on Cr(VI removal capacity in liquid medium

    Directory of Open Access Journals (Sweden)

    Avramović Nataša S.

    2013-01-01

    Full Text Available Pseudomonas aeruginosa NCAIM(P, B001380, a propitious bacterial strain isolated from mineral cutting oil was identified to be chromium tolerant and a producer of biosurfactant rhamnolipid (RL with potential application in heavy metal bioremediation. Culture growth, RL production and Cr(VI removal capacity of the strain in the presence of 50 mg L-1 (I and 100 mg L-1 of Cr(VI (II were studied. Maximum of RL production were found in the late-stationary phase at 72 h for both Cr(VI-amended cultures: I (236 mg L-1 and II (160 mg L-1, as well as the maximum of Cr(VI removal capacity: 70 % (I and 57 % (II. The amount of Cr in RL preparation II was 22 mg mg-1 determined by flame atomic absorption spectroscopy (FAAS. Appearance of a new band at 914 cm-1 in infrared (IR spectrum of RL (II indicated a significant proof for possible coordination of CrO42-ion with RL. The effect of Cr(VI on monorhamnolipids (RL1 and dirhamnolipids (RL2 distribution and its ratio were studied by electrospray ionization mass spectrometry (ESI-MS. An increase was observed in a RL2/RL1 ratio for II compared to control.

  10. Conditions for effective removal of pyrene from an artificially contaminated soil using Pseudomonas aeruginosa 57SJ rhamnolipids

    Energy Technology Data Exchange (ETDEWEB)

    Bordas, Francois [Institut national de la recherche scientifique, Centre INRS-Eau-Terre-Environnement, Universite du Quebec, 2800 rue Einstein, C.P. 7500, Sainte-Foy, Quebec, G1V 4C7 (Canada)]. E-mail: francois.bordas@unilim.fr; Lafrance, Pierre [Institut national de la recherche scientifique, Centre Eau, Terre et Environnement, Universite du Quebec, 490, rue de la Couronne, Quebec, G1K 9A9 (Canada)]. E-mail: pierre_lafrance@inrs-ete.uquebec.ca; Villemur, Richard [Institut national de la recherche scientifique, Centre INRS-Institut Armand Frappier, Universite du Quebec, 531 bd des Prairies, Laval, Quebec, H7V 1B7 (Canada)]. E-mail: richard.villemur@inrs-iaf.uquebec.ca

    2005-11-15

    The efficacy of a new rhamnolipid biosurfactants mixture to enhance the removal of pyrene from a soil artificially contaminated was investigated. The molar solubilization ratio (MSR) and the partition coefficient between the micelles and water (log K {sub m}) were found to be 7.5 x 10{sup -3} and 5.7, respectively. From soil column studies, the pyrene removal increased linearly with the concentration of the injected biosurfactants solution above the effective critical micellar concentration (0.4 g L{sup -1}). Flushing with a 5.0 g L{sup -1} biosurfactants solution increased the pyrene concentration in the effluent by 178 times. At high biosurfactants' concentrations (2.5 and 5.0 g L{sup -1}), the cumulative pyrene recovery reached 70%. This pyrene remobilization takes place independently of the soil organic carbon solubilization. This study provides a combination of batch and column experiments in order to find the conditions for effective soil remediation using a new rhamnolipids mixture. - The potential of newly isolated biosurfactants to mobilize PAHs from contaminated soils was evaluated from the determination in dynamic conditions of their effective critical micellar concentration.

  11. Lipopolysaccharide (LPS) inner-core phosphates are required for complete LPS synthesis and transport to the outer membrane in Pseudomonas aeruginosa PAO1.

    Science.gov (United States)

    Delucia, Angela M; Six, David A; Caughlan, Ruth E; Gee, Patricia; Hunt, Ian; Lam, Joseph S; Dean, Charles R

    2011-01-01

    also caused gross changes in cell morphology and led to the accumulation of an aberrant LPS lacking several core sugars and all core phosphates. The aberrant LPS failed to reach the OM, suggesting that WaaP is essential in P. aeruginosa because it is required to produce the full-length LPS that is recognized by the OM transport/assembly machinery in this organism. Therefore, WaaP may constitute a good target for the development of novel antipseudomonal agents.

  12. 30 CFR 772.11 - Notice requirements for exploration removing 250 tons of coal or less.

    Science.gov (United States)

    2010-07-01

    ... 250 tons of coal or less. 772.11 Section 772.11 Mineral Resources OFFICE OF SURFACE MINING RECLAMATION... requirements for exploration removing 250 tons of coal or less. (a) Any person who intends to conduct coal exploration operations outside a permit area during which 250 tons or less of coal will be removed,...

  13. Oligoribonuclease is the primary degradative enzyme for pGpG in Pseudomonas aeruginosa that is required for cyclic-di-GMP turnover.

    Science.gov (United States)

    Orr, Mona W; Donaldson, Gregory P; Severin, Geoffrey B; Wang, Jingxin; Sintim, Herman O; Waters, Christopher M; Lee, Vincent T

    2015-09-01

    The bacterial second messenger cyclic di-GMP (c-di-GMP) controls biofilm formation and other phenotypes relevant to pathogenesis. Cyclic-di-GMP is synthesized by diguanylate cyclases (DGCs). Phosphodiesterases (PDE-As) end signaling by linearizing c-di-GMP to 5'-phosphoguanylyl-(3',5')-guanosine (pGpG), which is then hydrolyzed to two GMP molecules by yet unidentified enzymes termed PDE-Bs. We show that pGpG inhibits a PDE-A from Pseudomonas aeruginosa. In a dual DGC and PDE-A reaction, excess pGpG extends the half-life of c-di-GMP, indicating that removal of pGpG is critical for c-di-GMP homeostasis. Thus, we sought to identify the PDE-B enzyme(s) responsible for pGpG degradation. A differential radial capillary action of ligand assay-based screen for pGpG binding proteins identified oligoribonuclease (Orn), an exoribonuclease that hydrolyzes two- to five-nucleotide-long RNAs. Purified Orn rapidly converts pGpG into GMP. To determine whether Orn is the primary enzyme responsible for degrading pGpG, we assayed cell lysates of WT and ∆orn strains of P. aeruginosa PA14 for pGpG stability. The lysates from ∆orn showed 25-fold decrease in pGpG hydrolysis. Complementation with WT, but not active site mutants, restored hydrolysis. Accumulation of pGpG in the ∆orn strain could inhibit PDE-As, increasing c-di-GMP concentration. In support, we observed increased transcription from the c-di-GMP-regulated pel promoter. Additionally, the c-di-GMP-governed auto-aggregation and biofilm phenotypes were elevated in the ∆orn strain in a pel-dependent manner. Finally, we directly detect elevated pGpG and c-di-GMP in the ∆orn strain. Thus, we identified that Orn serves as the primary PDE-B enzyme that removes pGpG, which is necessary to complete the final step in the c-di-GMP degradation pathway.

  14. Pseudomonas aeruginosa PA1006, which plays a role in molybdenum homeostasis, is required for nitrate utilization, biofilm formation, and virulence.

    Directory of Open Access Journals (Sweden)

    Melanie J Filiatrault

    Full Text Available Pseudomonas aeruginosa (Pae is a clinically important opportunistic pathogen. Herein, we demonstrate that the PA1006 protein is critical for all nitrate reductase activities, growth as a biofilm in a continuous flow system, as well as virulence in mouse burn and rat lung model systems. Microarray analysis revealed that ΔPA1006 cells displayed extensive alterations in gene expression including nitrate-responsive, quorum sensing (including PQS production, and iron-regulated genes, as well as molybdenum cofactor and Fe-S cluster biosynthesis factors, members of the TCA cycle, and Type VI Secretion System components. Phenotype Microarray™ profiles of ΔPA1006 aerobic cultures using Biolog plates also revealed a reduced ability to utilize a number of TCA cycle intermediates as well as a failure to utilize xanthine as a sole source of nitrogen. As a whole, these data indicate that the loss of PA1006 confers extensive changes in Pae metabolism. Based upon homology of PA1006 to the E. coli YhhP protein and data from the accompanying study, loss of PA1006 persulfuration and/or molybdenum homeostasis are likely the cause of extensive metabolic alterations that impact biofilm development and virulence in the ΔPA1006 mutant.

  15. Functional characterization of ExFadLO, an outer membrane protein required for exporting oxygenated long-chain fatty acids in Pseudomonas aeruginosa.

    Science.gov (United States)

    Martínez, Eriel; Estupiñán, Mónica; Pastor, F I Javier; Busquets, Montserrat; Díaz, Pilar; Manresa, Angeles

    2013-02-01

    Bacterial proteins of the FadL family have frequently been associated to the uptake of exogenous hydrophobic substrates. However, their outer membrane location and involvement in substrate uptake have been inferred mainly from sequence similarity to Escherichia coli FadL, the first well-characterized outer membrane transporters of Long-Chain Fatty Acids (LCFAs) in bacteria. Here we report the functional characterization of a Pseudomonas aeruginosa outer membrane protein (ORF PA1288) showing similarities to the members of the FadL family, for which we propose the name ExFadLO. We demonstrate herein that this protein is required to export LCFAs 10-HOME and 7,10-DiHOME, derived from a diol synthase oxygenation activity on oleic acid, from the periplasm to the extracellular medium. Accumulation of 10-HOME and 7,10-DiHOME in the extracellular medium of P. aeruginosa was abolished by a transposon insertion mutation in exFadLO (ExFadLO¯ mutant). However, intact periplasm diol synthase activity was found in this mutant, indicating that ExFadLO participates in the export of these oxygenated LCFAs across the outer membrane. The capacity of ExFadLO¯ mutant to export 10-HOME and 7,10-DiHOME was recovered after complementation with a wild-type, plasmid-expressed ExFadLO protein. A western blot assay with a variant of ExFadLO tagged with a V5 epitope confirmed the location of ExFadLO in the bacterial outer membrane under the experimental conditions tested. Our results provide the first evidence that FadL family proteins, known to be involved in the uptake of hydrophobic substrates from the extracellular environment, also function as secretion elements for metabolites of biological relevance.

  16. 76 FR 41434 - Removal of Certain Requirements Related to the Prescription Drug Marketing Act; Opportunity for...

    Science.gov (United States)

    2011-07-14

    ... prescription drug marketing and distribution. The primary purpose of the PDMA was to increase safeguards to... the Prescription Drug Marketing Act; Opportunity for Public Comment AGENCY: Food and Drug... remove a section of the Prescription Drug Marketing Act (PDMA) regulations requiring that prior to...

  17. Modeling regulatory policies associated with offshore structure removal requirements in the Gulf of Mexico

    Energy Technology Data Exchange (ETDEWEB)

    Kaiser, Mark J. [Center for Energy Studies, Louisiana State University, Energy Coast and Environment Building, Baton Rouge, LA (United States)

    2008-07-15

    Federal regulations require that a lease in the Outer Continental Shelf of the Gulf of Mexico be cleared of all structures within one year after production on the lease ceases, but in recent years, the Minerals Management Service has begun to encourage operators to remove idle (non-producing) structures on producing leases that are no longer ''economically viable''. At the end of 2003, there were 2175 producing structures, 898 idle (non-producing) structures, and 440 auxiliary (never-producing) structures on 1356 active leases; and 329 idle structures and 65 auxiliary structures on 273 inactive leases. The purpose of this paper is to model the impact of alternative regulatory policies on the removal trends of structures and the inventory of idle iron, and to provide first-order estimates of the cost of each regulatory option. A description of the modeling framework and implementation results is presented. (author)

  18. Comparison of the time required for removal of intraradicular cast posts using two Brazilian ultrasound devices

    Directory of Open Access Journals (Sweden)

    Manoel Brito-Júnior

    2009-03-01

    Full Text Available The aim of this in vitro study was to compare the time required for removal of intraradicular cast posts cemented with zinc phosphate (ZF or glass ionomer cement (GIC, using two Brazilian ultrasound devices (BUD. Seventy two human inferior premolars with single root canals were sectioned transversally at the cementoenamel junction. In each specimen, the root canal was endodontically treated, the post space was prepared to a depth of 9 mm and the canal was molded to obtain a post impression. After the casting procedures, the posts were randomly distributed into 2 groups (n = 36 according to the luting material used: G1 - ZF and G2 - GIC. The tooth and luted post set was then embedded in an acrylic resin block. The groups were then divided into 3 subgroups (n = 12 according to the ultrasound device used: A - Enac (Osada Electric, Japan, used as a control group; B - Profi II Ceramic (Dabi Atlante, Brazil and C - Jet Sonic Satelec (Gnatus, Brazil. The posts were submitted to the vibration process with maximum power set on all surrounding surfaces. Time of application was recorded with a chronometer until complete post dislodgment, and the data were analyzed by the ANOVA test (p < 0.05. The averages required for post removal in G1 and G2 were respectively 41.42 and 92.03 seconds, with significant statistical difference (p = 0.001. No statistical difference was observed among the three ultrasound devices (p = 0.088, and the BUD presented a performance similar to that of the international gold standard device (Enac. Moreover, the type of luting agent had a greater influence on the time required for post removal than the origin of the ultrasonic unit.

  19. Silver against Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Kirketerp-Møller, K.; Kristiansen, S.

    2007-01-01

    Silver has been recognized for its antimicrobial properties for centuries. Most studies on the antibacterial efficacy of silver, with particular emphasis on wound healing, have been performed on planktonic bacteria. Our recent studies, however, strongly suggest that colonization of wounds involves...... bacteria in both the planktonic and biofilm modes of growth. The action of silver on mature in vitro biofilms of Pseudomonas aeruginosa, a primary pathogen of chronic infected wounds, was investigated. The results show that silver is very effective against mature biofilms of P. aeruginosa......, but that the silver concentration is important. A concentration of 5-10 ig/mL silver sulfadiazine eradicated the biofilm whereas a lower concentration (1 ig/mL) had no effect. The bactericidal concentration of silver required to eradicate the bacterial biofilm was 10-100 times higher than that used to eradicate...

  20. Pseudomonas aeruginosa killing of Caenorhabditis elegans used to identify P. aeruginosa virulence factors

    Science.gov (United States)

    Tan, Man-Wah; Rahme, Laurence G.; Sternberg, Jeffrey A.; Tompkins, Ronald G.; Ausubel, Frederick M.

    1999-01-01

    We reported recently that the human opportunistic pathogen Pseudomonas aeruginosa strain PA14 kills Caenorhabditis elegans and that many P. aeruginosa virulence factors (genes) required for maximum virulence in mouse pathogenicity are also required for maximum killing of C. elegans. Here we report that among eight P. aeruginosa PA14 TnphoA mutants isolated that exhibited reduced killing of C. elegans, at least five also exhibited reduced virulence in mice. Three of the TnphoA mutants corresponded to the known virulence-related genes lasR, gacA, and lemA. Three of the mutants corresponded to known genes (aefA from Escherichia coli, pstP from Azotobacter vinelandii, and mtrR from Neisseria gonorrhoeae) that had not been shown previously to play a role in pathogenesis, and two of the mutants contained TnphoA inserted into novel sequences. These data indicate that the killing of C. elegans by P. aeruginosa can be exploited to identify novel P. aeruginosa virulence factors important for mammalian pathogenesis. PMID:10051655

  1. Pseudomonas aeruginosa: assessment of risk from drinking water.

    Science.gov (United States)

    Hardalo, C; Edberg, S C

    1997-01-01

    Pseudomonas aeruginosa is an ubiquitous environmental bacterium. It can be recovered, often in high numbers, in common food, especially vegetables. Moreover, it can be recovered in low numbers in drinking water. A small percentage of clones of P. aeruginosa possesses the required number of virulence factors to cause infection. However, P. aeruginosa will not proliferate on normal tissue but requires previously organs. Further narrowing the risk to human health is that only certain specific hosts are at risk, including patients with profound neutropenia, cystic fibrosis, severe burns, and those subject to foreign device installation. Other than these very well-defined groups, the general population is refractory to infection with P. aeruginosa. Because of its ubiquitous nature, it is not only not practical to eliminate P. aeruginosa from our food and drinking water, but attempts to do so would produce disinfection byproducts more hazardous than the species itself. Moreover, because there is no readily available sensitive and specific means to detect and identify P. aeruginosa available in the field, any potential regulation governing its control would not have a defined laboratory test measure of outcome. Accordingly, attempts to regulate P. aeruginosa in drinking water would not yield public health protection benefits and could, in fact, be counterproductive in this regard.

  2. 77 FR 59318 - Removal of 30-Day Residency Requirement for Per Diem Payments

    Science.gov (United States)

    2012-09-27

    ... able to provide stable nursing home care via State homes as we intend. Additionally, removing the 30... Regulation and Regulatory Review) emphasizes the importance of quantifying both costs and benefits, reducing... annual effect on the economy of $100 million or more or adversely affect in a material way the economy,...

  3. 76 FR 29991 - Live Goats and Swine for Export; Removal of Certain Testing Requirements

    Science.gov (United States)

    2011-05-24

    ... brucellosis testing of goats and breeding swine intended for export to countries that do not require such... brucellosis prior to export. Some countries do not require that goats and breeding swine be tested for... eliminate the requirement for pre-export tuberculosis and brucellosis testing of goats and breeding...

  4. 75 FR 56912 - Live Goats and Swine for Export; Removal of Certain Testing Requirements

    Science.gov (United States)

    2010-09-17

    ... requirement for pre-export tuberculosis and brucellosis testing of goats and breeding swine intended for... exportation be tested for tuberculosis and, for some goats, brucellosis prior to export. Section 91.9 requires... countries do not require that goats and breeding swine be tested for tuberculosis and brucellosis prior...

  5. Development of a Pseudomonas aeruginosa Agmatine Biosensor

    Directory of Open Access Journals (Sweden)

    Adam Gilbertsen

    2014-10-01

    Full Text Available Agmatine, decarboxylated arginine, is an important intermediary in polyamine production for many prokaryotes, but serves higher functions in eukaryotes such as nitric oxide inhibition and roles in neurotransmission. Pseudomonas aeruginosa relies on the arginine decarboxylase and agmatine deiminase pathways to convert arginine into putrescine. One of the two known agmatine deiminase operons, aguBA, contains an agmatine sensitive TetR promoter controlled by AguR. We have discovered that this promoter element can produce a titratable induction of its gene products in response to agmatine, and utilized this discovery to make a luminescent agmatine biosensor in P. aeruginosa. The genome of the P. aeruginosa lab strain UCBPP-PA14 was altered to remove both its ability to synthesize or destroy agmatine, and insertion of the luminescent reporter construct allows it to produce light in proportion to the amount of exogenous agmatine applied from ~100 nM to 1mM. Furthermore it does not respond to related compounds including arginine or putrescine. To demonstrate potential applications the biosensor was used to detect agmatine in spent supernatants, to monitor the development of arginine decarboxylase over time, and to detect agmatine in the spinal cords of live mice.

  6. Risk assessment of Pseudomonas aeruginosa in water.

    Science.gov (United States)

    Mena, Kristina D; Gerba, Charles P

    2009-01-01

    . 1986). Outbreaks have also been reported from exposure to P. aeruginosa in swimming pools and water slides. Although P. aeruginosa has a reputation for being resistant to disinfection, most studies show that it does not exhibit any marked resistance to the disinfectants used to treat drinking water such as chlorine, chloramines, ozone, or iodine. One author, however, did find it to be slightly more resistant to UV disinfection than most other bacteria (Wolfe 1990). Although much has been written about biofilms in the drinking water industry, very little has been reported regarding the role of P. aeruginosa in biofilms. Tap water appears to be a significant route of transmission in hospitals, from colonization of plumbing fixtures. It is still not clear if the colonization results from the water in the distribution system, or personnel use within the hospital. Infections and colonization can be significantly reduced by placement of filters on the water taps. The oral dose of P. aeruginosa required to establish colonization in a healthy subject is high (George et al. 1989a). During dose-response studies, even when subjects (mice or humans) were colonized via ingestion, there was no evidence of disease. P. aeruginosa administered by the aerosol route at levels of 10(7) cells did cause disease symptoms in mice, and was lethal in aerosolized doses of 10(9) cells. Aerosol dose-response studies have not been undertaken with human subjects. Human health risks associated with exposure to P. aeruginosa via drinking water ingestion were estimated using a four-step risk assessment approach. The risk of colonization from ingesting P. aeruginosa in drinking water is low. The risk is slightly higher if the subject is taking an antibiotic resisted by P. aeruginosa. The fact that individuals on ampicillin are more susceptible to Pseudomonas gastrointestinal infection probably results from suppression of normal intestinal flora, which would allow Pseudomonas to colonize. The process of

  7. The galactophilic lectin (PA-IL, gene LecA) from Pseudomonas aeruginosa. Its binding requirements and the localization of lectin receptors in various mouse tissues

    DEFF Research Database (Denmark)

    Kirkeby, Svend; Hansen, Axel K; d'Apice, Anthony

    2006-01-01

    . It is concluded that the carbohydrate recognizing site of the lectin can have a binding requirement of only one saccharide. Lectin histochemistry was performed on sections from wild type mice and from knock-out mice, which lack function of the alpha.1,3-galactosyltransferase gene. All assays with the P...

  8. Crystal Structure of the Pseudomonas aeruginosa Virulence Factor Regulator

    Energy Technology Data Exchange (ETDEWEB)

    Cordes, Timothy J.; Worzalla, Gregory A.; Ginster, Aaron M.; Forest, Katrina T. (UW)

    2012-09-07

    Virulence factor regulator (Vfr) enhances Pseudomonas aeruginosa pathogenicity through its role as a global transcriptional regulator. The crystal structure of Vfr shows that it is a winged-helix DNA-binding protein like its homologue cyclic AMP receptor protein (CRP). In addition to an expected primary cyclic AMP-binding site, a second ligand-binding site is nestled between the N-terminal domain and the C-terminal helix-turn-helix domain. Unlike CRP, Vfr is a symmetric dimer in the absence of DNA. Removal of seven disordered N-terminal residues of Vfr prvents the growth of P. aeruginosa.

  9. The Smc5-Smc6 complex is required to remove chromosome junctions in meiosis.

    Directory of Open Access Journals (Sweden)

    Sarah Farmer

    Full Text Available Meiosis, a specialized cell division with a single cycle of DNA replication round and two consecutive rounds of nuclear segregation, allows for the exchange of genetic material between parental chromosomes and the formation of haploid gametes. The structural maintenance of chromosome (SMC proteins aid manipulation of chromosome structures inside cells. Eukaryotic SMC complexes include cohesin, condensin and the Smc5-Smc6 complex. Meiotic roles have been discovered for cohesin and condensin. However, although Smc5-Smc6 is known to be required for successful meiotic divisions, the meiotic functions of the complex are not well understood. Here we show that the Smc5-Smc6 complex localizes to specific chromosome regions during meiotic prophase I. We report that meiotic cells lacking Smc5-Smc6 undergo catastrophic meiotic divisions as a consequence of unresolved linkages between chromosomes. Surprisingly, meiotic segregation defects are not rescued by abrogation of Spo11-induced meiotic recombination, indicating that at least some chromosome linkages in smc5-smc6 mutants originate from other cellular processes. These results demonstrate that, as in mitosis, Smc5-Smc6 is required to ensure proper chromosome segregation during meiosis by preventing aberrant recombination intermediates between homologous chromosomes.

  10. A risk assessment of Pseudomonas aeruginosa in swimming pools: a review.

    Science.gov (United States)

    Rice, Scott A; van den Akker, Ben; Pomati, Francesco; Roser, David

    2012-06-01

    Despite routine monitoring and disinfection, treated swimming pools are frequently contaminated with the opportunistic pathogen Pseudomonas aeruginosa, which can represent a significant public health threat. This review was undertaken to identify the current understanding of risk factors associated with pool operation with respect to P. aeruginosa. The ecology and factors that promote growth of P. aeruginosa in the pool environment are complex and dynamic and so we applied a systematic risk assessment approach to integrate existing data, with the aim to improve pool management and safety. Sources of P. aeruginosa, types of infections, dose responses, routes of transmission, as well as the efficacy of current disinfectant treatments were reviewed. This review also highlights the critical knowledge gaps that are required for a more robust, quantitative risk assessment of P. aeruginosa. Quantitative risk management strategies have been successfully applied to drinking water systems and should similarly be amenable to developing a better understanding of the risk posed by P. aeruginosa in swimming pools.

  11. Pseudomonas aeruginosa biofilm infections

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2014-01-01

    Bacteria in natural, industrial and clinical settings predominantly live in biofilms, i.e., sessile structured microbial communities encased in self-produced extracellular matrix material. One of the most important characteristics of microbial biofilms is that the resident bacteria display...... a remarkable increased tolerance toward antimicrobial attack. Biofilms formed by opportunistic pathogenic bacteria are involved in devastating persistent medical device-associated infections, and chronic infections in individuals who are immune-compromised or otherwise impaired in the host defense. Because...... the use of conventional antimicrobial compounds in many cases cannot eradicate biofilms, there is an urgent need to develop alternative measures to combat biofilm infections. The present review is focussed on the important opportunistic pathogen and biofilm model organism Pseudomonas aeruginosa. Initially...

  12. Pseudomonas aeruginosa in Healthcare Settings

    Science.gov (United States)

    ... Address What's this? Submit What's this? Submit Button Pseudomonas aeruginosa in Healthcare Settings Recommend on Facebook Tweet ... and/or help treat infections? What is a Pseudomonas infection? Pseudomonas infection is caused by strains of ...

  13. HD-GYP domain proteins regulate biofilm formation and virulence in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Ryan, Robert P.; Lucey, Jean; O'Donovan, Karen

    2009-01-01

    2572 had a negative influence on swarming that was cryptic and was revealed only after removal of an uncharacterized C-terminal domain. Mutation of PA4108, PA4781 and PA2572 had distinct effects on biofilm formation and architecture of P. aeruginosa. All three proteins contributed to virulence of P...... residues (YN-GYP). Here we have investigated the role of these proteins in biofilm formation, virulence factor synthesis and virulence of P. aeruginosa. Mutation of PA4108 and PA4781 led to an increase in the level of cyclic-di-GMP in P. aeruginosa, consistent with the predicted activity of the encoded...

  14. Chronic Pseudomonas aeruginosa cervical osteomyelitis

    Directory of Open Access Journals (Sweden)

    Sujeet Kumar Meher

    2016-01-01

    Full Text Available Pseudomonas aeruginosa is a rare cause of osteomyelitis of the cervical spine and is usually seen in the background of intravenous drug use and immunocompromised state. Very few cases of osteomyelitis of the cervical spine caused by pseudomonas aeruginosa have been reported in otherwise healthy patients. This is a case presentation of a young female, who in the absence of known risk factors for cervical osteomyelitis presented with progressively worsening neurological signs and symptoms.

  15. Autocatalytic activation of the furin zymogen requires removal of the emerging enzyme's N-terminus from the active site.

    Directory of Open Access Journals (Sweden)

    Katarzyna Gawlik

    Full Text Available Before furin can act on protein substrates, it must go through an ordered process of activation. Similar to many other proteinases, furin is synthesized as a zymogen (profurin which becomes active only after the autocatalytic removal of its auto-inhibitory prodomain. We hypothesized that to activate profurin its prodomain had to be removed and, in addition, the emerging enzyme's N-terminus had to be ejected from the catalytic cleft.We constructed and analyzed the profurin mutants in which the egress of the emerging enzyme's N-terminus from the catalytic cleft was restricted. Mutants were autocatalytically processed at only the primary cleavage site Arg-Thr-Lys-Arg(107 downward arrowAsp(108, but not at both the primary and the secondary (Arg-Gly-Val-Thr-Lys-Arg(75 downward arrowSer(76 cleavage sites, yielding, as a result, the full-length prodomain and mature furins commencing from the N-terminal Asp108. These correctly processed furin mutants, however, remained self-inhibited by the constrained N-terminal sequence which continuously occupied the S' sub-sites of the catalytic cleft and interfered with the functional activity. Further, using the in vitro cleavage of the purified prodomain and the analyses of colon carcinoma LoVo cells with the reconstituted expression of the wild-type and mutant furins, we demonstrated that a three-step autocatalytic processing including the cleavage of the prodomain at the previously unidentified Arg-Leu-Gln-Arg(89 downward arrowGlu(90 site, is required for the efficient activation of furin.Collectively, our results show the restrictive role of the enzyme's N-terminal region in the autocatalytic activation mechanisms. In a conceptual form, our data apply not only to profurin alone but also to a range of self-activated proteinases.

  16. Antibacterial, anti-swarming and anti-biofilm formation activities of Chamaemelum nobile against Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Hossein Kazemian

    2015-08-01

    Full Text Available AbstractINTRODUCTION:Chamomile ( Chamaemelum nobile is widely used throughout the world, and has anti-inflammatory, deodorant, bacteriostatic, antimicrobial, carminative, sedative, antiseptic, anti-catarrhal, and spasmolytic properties. Because of the increasing incidence of drug-resistant bacteria, the development of natural antibacterial sources such as medical herbs for the treatment of infectious diseases is necessary. Extracts from different plant parts such as the leaves, flowers, fruit, and bark of Combretum albiflorum, Laurus nobilis , and Sonchus oleraceus were found to possess anti-quorum sensing (QS activities. In this study, we evaluated the effect of C. nobile against Pseudomonas aeruginosa biofilm formationMETHODS:The P. aeruginosa samples were isolated from patients with different types of infection, including wound infection, septicemia, and urinary tract infection. The flowers of C. nobile were dried and the extract was removed using a rotary device and then dissolved in dimethyl sulfoxide at pH 7.4. The microdilution method was used to evaluate the minimum inhibitory concentration (MIC of this extract on P. aeruginosa , and biofilm inhibition was assayed.RESULTS:Eighty percent of the isolated samples (16/20 could form a biofilm, and most of these were isolated from wound infections. The biofilm inhibitory concentration of the C. nobile extract was 6.25-25mg/ml, whereas the MIC was 12.5-50mg/ml.CONCLUSIONS:The anti-QS property of C. nobile may play an important role in its antibacterial activity, thus offering an additional strategy in the fight against bacterial infections. However, molecular investigation is required to explore the exact mechanisms of the antibacterial action and functions of this phytocompound.

  17. Light and Phosphate Competition Between Cylindrospermopsis raciborskii and Microcystis aeruginosa is Strain Dependent

    NARCIS (Netherlands)

    Marinho, M.M.; Gonçalves Souza, M.B.; Lürling, M.

    2013-01-01

    The hypothesis that outcomes of phosphorus and light competition between Cylindrospermopsis raciborskii and Microcystis aeruginosa are strain dependent was tested experimentally. Critical requirements of phosphorus (P*) and of light (I*) of two strains of each species were determined through monocul

  18. ARSENIC DEGRADATION BY Pseudomonas aeruginosa FOR WATER BIOREMEDIATION. PRELIMINARY STUDY

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    Esther E. Pellizzari

    2015-03-01

    Full Text Available The aim of this study was to investigate the arsenic resistance in pure cultivations of Pseudomonas aeruginosa isolated from Presidencia Roque Sáenz Peña groundwater (Chaco province, and evaluate the possibility of its use to remove arsenic from groundwater. Strains were immobilized in natural stone and cultivated in salts broth and 1 mgAs/L. The arsenic resistance and biofilm formation were observed, obtaining interaction between cells, rock and arsenic. Arsenic removal was evaluated during 3 months and its final percentage of the experiment was 60%.

  19. Maturation of the cytochrome cd 1 nitrite reductase NirS from Pseudomonas aeruginosa requires transient interactions between the three proteins NirS, NirN and NirF

    OpenAIRE

    2013-01-01

    The periplasmic cytochrome cd 1 nitrite reductase NirS occurring in denitrifying bacteria such as the human pathogen Pseudomonas aeruginosa contains the essential tetrapyrrole cofactors haem c and haem d 1. Whereas the haem c is incorporated into NirS by the cytochrome c maturation system I, nothing is known about the insertion of the haem d 1 into NirS. Here, we show by co-immunoprecipitation that NirS interacts with the potential haem d 1 insertion protein NirN in vivo. This NirS–NirN inter...

  20. Microevolution of Pseudomonas aeruginosa to a chronic pathogen of the cystic fibrosis lung.

    Science.gov (United States)

    Hogardt, Michael; Heesemann, Jürgen

    2013-01-01

    Pseudomonas aeruginosa is the leading pathogen of chronic cystic fibrosis (CF) lung infection. Life-long persistance of P. aeruginosa in the CF lung requires a sophisticated habitat-specific adaptation of this pathogen to the heterogeneous and fluctuating lung environment. Due to the high selective pressure of inflamed CF lungs, P. aeruginosa increasingly experiences complex physiological and morphological changes. Pulmonary adaptation of P. aeruginosa is mediated by genetic variations that are fixed by the repeating interplay of mutation and selection. In this context, the emergence of hypermutable phenotypes (mutator strains) obviously improves the microevolution of P. aeruginosa to the diverse microenvironments of the CF lung. Mutator phenotypes are amplified during CF lung disease and accelerate the intraclonal diversification of P. aeruginosa. The resulting generation of numerous subclonal variants is advantegous to prepare P. aeruginosa population for unpredictable stresses (insurance hypothesis) and thus supports long-term survival of this pathogen. Oxygen restriction within CF lung environment further promotes persistence of P. aeruginosa due to increased antibiotic tolerance, alginate production and biofilm formation. Finally, P. aeruginosa shifts from an acute virulent pathogen of early infection to a host-adapted chronic virulent pathogen of end-stage infection of the CF lung. Common changes that are observed among chronic P. aeruginosa CF isolates include alterations in surface antigens, loss of virulence-associated traits, increasing antibiotic resistances, the overproduction of the exopolysaccharide alginate and the modulation of intermediary and micro-aerobic metabolic pathways (Hogardt and Heesemann, Int J Med Microbiol 300(8):557-562, 2010). Loss-of-function mutations in mucA and lasR genes determine the transition to mucoidity and loss of quorum sensing, which are hallmarks of the chronic virulence potential of P. aeruginosa. Metabolic factors

  1. Necessity of 3D visualization for the removal of lower wisdom teeth: required sample size to prove non-inferiority of panoramic radiography compared to CBCT.

    Science.gov (United States)

    Roeder, Felix; Wachtlin, Daniel; Schulze, Ralf

    2012-06-01

    The availability of cone beam computed tomography (CBCT) and the numbers of CBCT scans rise constantly, increasing the radiation burden to the patient. A growing discussion is noticeable if a CBCT scan prior to the surgical removal of wisdom teeth may be indicated. We aimed to confirm non-inferiority with respect to damage of the inferior alveolar nerve in patients diagnosed by panoramic radiography compared to CBCT in a prospective randomized controlled multicentre trial. Sample size (number of required third molar removals) was calculated for the study and control groups as 183,474 comparing temporary and 649,036 comparing permanent neurosensory disturbances of the inferior alveolar nerve. Modifying parameter values resulted in sample sizes ranging from 39,584 to 245,724 respectively 140,024 to 869,250. To conduct a clinical study to prove a potential benefit from CBCT scans prior to surgical removal of lower wisdom teeth with respect to the most important parameter, i.e., nerval damage, is almost impossible due to the very large sample sizes required. This fact vice versa indicates that CBCT scans should only be performed in high risk wisdom tooth removals.

  2. A functional type I topoisomerase from Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Roper Benjamin J

    2009-03-01

    Full Text Available Abstract Background Pseudomonas aeruginosa encodes a putative topoisomerase with sequence similarity to the eukaryotic type IB topoisomerase from Vaccinia virus. Residues in the active site are conserved, notably Tyr292 which would be predicted to form the transient covalent bond to DNA. Results The gene encoding the P. aeruginosa topoisomerase I was cloned and expressed in E. coli. The enzyme relaxes supercoiled DNA, while a mutant containing a Tyr292 to Phe substitution at the active site was found to be catalytically inert. This is consistent with the role of Tyr in forming the covalent intermediate. Like Vaccinia topoisomerase, the P. aeruginosa topoisomerase relaxes DNA in the absence of ATP, but unlike Vaccinia topoisomerase, P. aeruginosa topoisomerase does not relax supercoiled DNA without MgCl2 present. In addition, high concentration of NaCl is not able to substitute for MgCl2 as seen for Vaccinia topoisomerase. A truncated derivative of the topoisomerase lacking residues 1–98 relaxes DNA, with both full length and truncated enzyme exhibiting equivalent requirements for divalent cations and the ability to relax DNA to completion, suggesting a shared domain organization. DNA-binding assays suggest an only modest preference for the CCCTT pentameric sequence required for transesterification by Vaccinia topoisomerase IB. Conclusion P. aeruginosa encodes a functional topoisomerase with significant similarity to the type IB enzyme encoded by poxviruses. In contrast to the Vaccinia-encoded homolog, the P. aeruginosa-encoded enzyme requires divalent cations for catalytic activity, relaxes DNA to completion, and does not exhibit a strong preference for the pentameric sequence stringently required by the Vaccinia-encoded homolog. A comparison with the structure of poxviral topoisomerase in complex with DNA suggests that bacterial homologs of the eukaryotic type IB topoisomerase may exhibit a relaxed sequence preference due to the lack of

  3. Gallium-Protoporphyrin IX Inhibits Pseudomonas aeruginosa Growth by Targeting Cytochromes

    Science.gov (United States)

    Hijazi, Sarah; Visca, Paolo; Frangipani, Emanuela

    2017-01-01

    Pseudomonas aeruginosa is a challenging pathogen due to both innate and acquired resistance to antibiotics. It is capable of causing a variety of infections, including chronic lung infection in cystic fibrosis (CF) patients. Given the importance of iron in bacterial physiology and pathogenicity, iron-uptake and metabolism have become attractive targets for the development of new antibacterial compounds. P. aeruginosa can acquire iron from a variety of sources to fulfill its nutritional requirements both in the environment and in the infected host. The adaptation of P. aeruginosa to heme iron acquisition in the CF lung makes heme utilization pathways a promising target for the development of new anti-Pseudomonas drugs. Gallium [Ga(III)] is an iron mimetic metal which inhibits P. aeruginosa growth by interfering with iron-dependent metabolism. The Ga(III) complex of the heme precursor protoporphyrin IX (GaPPIX) showed enhanced antibacterial activity against several bacterial species, although no inhibitory effect has been reported on P. aeruginosa. Here, we demonstrate that GaPPIX is indeed capable of inhibiting the growth of clinical P. aeruginosa strains under iron-deplete conditions, as those encountered by bacteria during infection, and that GaPPIX inhibition is reversed by iron. Using P. aeruginosa PAO1 as model organism, we show that GaPPIX enters cells through both the heme-uptake systems has and phu, primarily via the PhuR receptor which plays a crucial role in P. aeruginosa adaptation to the CF lung. We also demonstrate that intracellular GaPPIX inhibits the aerobic growth of P. aeruginosa by targeting cytochromes, thus interfering with cellular respiration.

  4. Aerobic biodegradation pathway for Remazol Orange by Pseudomonas aeruginosa.

    Science.gov (United States)

    Sarayu, K; Sandhya, S

    2010-02-01

    Removal of azo dyes from effluent generated by textile industries is rather difficult. Azo dyes represent a major class of synthetic colorants that are mutagenic and carcinogenic. Pseudomonas aeruginosa grew well in the presence of Remazol Orange (RO) and was able to decolorize and degrade it. In the present study, the decolorization and degradation efficiency using single culture P. aeruginosa with RO and textile wastewaters is studied. The elucidation of decolorization pathway for P. aeruginosa is of special interest. The degradation pathway and the metabolic products formed during the degradation were also predicted with the help of high performance liquid chromatography, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy analysis. The data show the cleavage of the azo dye RO to form both methyl metanilic acid and 4-aminobenzoic acid after decolorization and finally to oxidation forms benzoic acid, alkenes, aldehydes, and alkynes. The organism was able to decolorize the dye RO and wastewater effectively to the maximum of 82.4% and 62%, respectively.

  5. An unusual presentation of Pseudomonas aeruginosa blebitis following combined surgery

    Directory of Open Access Journals (Sweden)

    Shabana Bharathi

    2014-01-01

    Full Text Available We report a case of blebitis that occurred 3 years later following a combined glaucoma and cataract surgery. It was an atypical presentation, as patient had no classical fiery looking signs of blebitis despite the isolated organism being Pseudomonas aeruginosa. Improvized surgical techniques like use of Mitomycin C, releasable flap sutures though considered as part of the recommended procedure for better surgical outcomes, their role as potential risk factors for visually blinding complications like endophthalmitis are often overlooked. This case report throws light on such risk factors for bleb associated infections and recommends removal or trimming of all releasable sutures and the need for a regular postoperative follow-up.

  6. Low-flow CO2 removal integrated into a renal-replacement circuit can reduce acidosis and decrease vasopressor requirements

    Science.gov (United States)

    2013-01-01

    Introduction Lung-protective ventilation in patients with ARDS and multiorgan failure, including renal failure, is often paralleled with a combined respiratory and metabolic acidosis. We assessed the effectiveness of a hollow-fiber gas exchanger integrated into a conventional renal-replacement circuit on CO2 removal, acidosis, and hemodynamics. Methods In ten ventilated critically ill patients with ARDS and AKI undergoing renal- and respiratory-replacement therapy, effects of low-flow CO2 removal on respiratory acidosis compensation were tested by using a hollow-fiber gas exchanger added to the renal-replacement circuit. This was an observational study on safety, CO2-removal capacity, effects on pH, ventilator settings, and hemodynamics. Results CO2 elimination in the low-flow circuit was safe and was well tolerated by all patients. After 4 hours of treatment, a mean reduction of 17.3 mm Hg (−28.1%) pCO2 was observed, in line with an increase in pH. In hemodynamically instable patients, low-flow CO2 elimination was paralleled by hemodynamic improvement, with an average reduction of vasopressors of 65% in five of six catecholamine-dependent patients during the first 24 hours. Conclusions Because no further catheters are needed, besides those for renal replacement, the implementation of a hollow-fiber gas exchanger in a renal circuit could be an attractive therapeutic tool with only a little additional trauma for patients with mild to moderate ARDS undergoing invasive ventilation with concomitant respiratory acidosis, as long as no severe oxygenation defects indicate ECMO therapy. PMID:23883472

  7. The removal of the consideration requirement, and the consequent clarification on duress, for verbal modifications to Irish construction contracts

    OpenAIRE

    Mooney, Conor Francis Joseph

    2015-01-01

    peer-reviewed This thesis studies the consideration requirement for verbally-made contract variations in Irish construction contracts, and proposes how this specific circumstance can be best served by the law of contract. Construction is a complex and uncertain industry; and an effective construction industry is important and necessary for Ireland: it is a key driver of a functioning, developed economy, and it requires a sustainable level of activity to maintain infrastructu...

  8. Minimization of steam requirements and enhancement of water-gas shift reaction with warm gas temperature CO2 removal

    Science.gov (United States)

    Siriwardane, Ranjani V; Fisher, II, James C

    2013-12-31

    The disclosure utilizes a hydroxide sorbent for humidification and CO.sub.2 removal from a gaseous stream comprised of CO and CO.sub.2 prior to entry into a water-gas-shift reactor, in order to decrease CO.sub.2 concentration and increase H.sub.2O concentration and shift the water-gas shift reaction toward the forward reaction products CO.sub.2 and H.sub.2. The hydroxide sorbent may be utilized for absorbtion of CO.sub.2 exiting the water-gas shift reactor, producing an enriched H.sub.2 stream. The disclosure further provides for regeneration of the hydroxide sorbent at temperature approximating water-gas shift conditions, and for utilizing H.sub.2O product liberated as a result of the CO.sub.2 absorption.

  9. Control of the harmful alga Microcystis aeruginosa and absorption of nitrogen and phosphorus by Candida utilis.

    Science.gov (United States)

    Kong, Yun; Xu, Xiangyang; Zhu, Liang; Miao, Lihong

    2013-01-01

    This study is aimed at controlling eutrophication through converting the nutrients such as nitrogen and phosphorus into microbial protein and simultaneously inhibiting the growth of Microcystis aeruginosa by Candida utilis. C. utilis and M. aeruginosa (initial cell density was 2.25 × 10(7) and 4.15 × 10(7) cells·mL(-1)) were cultured together in the absence or presence of a carbon source (glucose) during a 10-day experiment. In the absence of carbon source, the measured removal efficiencies of NH(4) (+)-N and PO(4) (3-)-P were 41.39 ± 2.19 % and 82.93 ± 3.95 %, respectively, at the second day, with the removal efficiency of 67.82 ± 2.29 % for M. aeruginosa at the fourth day. In contrast, the removal efficiencies of NH(4) (+)-N and PO(4) (3-)-P were increased to 87.45 ± 4.25 % and 83.73 ± 3.55 %, respectively, while the removal efficiency of M. aeruginosa decreased to 37.89 ± 8.41 % in the presence of the carbon source (C/N = 2:1). These results showed that the growth of M. aeruginosa was inhibited by C. utilis. Our finding sheds light on a novel potential approach for yeast to consume nutrients and control harmful algal during bloom events.

  10. 8 CFR 1216.5 - Waiver of requirement to file joint petition to remove conditions by alien spouse.

    Science.gov (United States)

    2010-01-01

    .... (b) Fee. Form I-751 shall be accompanied by the appropriate fee required under § 103.7(b) of 8 CFR... the field. An evaluation which was obtained in the course of the divorce proceedings may be submitted.... If the decision is adverse, the director shall advise the alien of the reasons therefor, notify...

  11. Use of the paraffin wax baiting system for identification of Pseudomonas aeruginosa clinical isolates.

    Science.gov (United States)

    Massengale, A R; Ollar, R A; Giordano, S J; Felder, M S; Aronoff, S C

    1999-11-01

    Pseudomonas aeruginosa is the primary pathogen among the Pseudomonads and is known for its minimal nutritional requirements, capacity to use paraffin as a sole carbon source, and biofilm formation. Because the ability of Pseudomonads to grow on paraffin is not commonly found among human pathogens and the primary Pseudomonas human pathogen is P. aeruginosa, we studied the adaptation of the paraffin baiting system for the growth and identification of clinical isolates of P. aeruginosa. We also studied the effectiveness of combining a fluorescence assay measuring fluorescein (pyoverdin) production and oxidase test with the paraffin baiting assay for P. aeruginosa speciation. Strains were tested for the capacity to use paraffin as a sole carbon source using the paraffin baiting system with Czapek's minimal salt medium. Of 111 P. aeruginosa clinical isolates tested for using paraffin as a sole carbon source, 45% exhibited growth on paraffin at 24 h and 76.6% exhibited growth on paraffin at 48 h. The ability of the reference strains and clinical isolates were then tested for their ability to associate with the paraffin slide in the presence of an additional carbon source. Of 111 P. aeruginosa clinical isolates tested, 85 strains (76.6%), and 102 (93%) were associated with the paraffin surface at 24 and 48 h. We successfully combined fluorescence and oxidase assays with the paraffin baiting system for identification of P. aeruginosa. The simple and inexpensive paraffin baiting system is a useful method for the identification and study of P. aeruginosa suitable for both the clinical and research laboratory.

  12. Effect of environmental factors on allelopathic inhibition of Microcystis aeruginosa by berberine.

    Science.gov (United States)

    Zhang, Shulin; Dai, Wei; Bi, Xiangdong; Zhang, Dajuan; Xing, Kezhi

    2013-01-01

    To understand how environmental conditions affect the allelopathic inhibition of toxic Microcystis aeruginosa by berberine, the independent effects of some environmental factors, including temperature, light, and aeration, on the growth and extracellular microcystin (MC) content of M. aeruginosa (FACHB 905) treated with 0.000 and 0.001% (w/v) berberine were investigated. The results showed that higher temperature and light density, and aeration in daytime were beneficial for the growth of M. aeruginosa under the measured environmental conditions. The allelopathic effects of berberine on M. aeruginosa were closely associated with the environmental conditions. Berberine had the best inhibitory effects when temperature, light and aeration were more optimal for growth. In darkness, no changes in the density of M. aeruginosa were observed with the prolongation of culture time and berberine could hardly exhibit algicidal effects. Disturbance in the photosynthesis process might be one of the main reasons responsible for algicidal function. Berberine could increase extracellular MC contents significantly via killing and lyzing algal cells. Other treatments coupled with berberine needed to be carried out to degrade or remove MC released from berberine-killed algal cells.

  13. Characterization of bacteriophages infecting clinical isolates of Pseudomonas aeruginosa stored in a culture collection

    Directory of Open Access Journals (Sweden)

    C.C.S. Zanetti

    2013-08-01

    Full Text Available Some clinical isolates of Pseudomonas aeruginosa stored in our culture collection did not grow or grew poorly and showed lysis on the culture plates when removed from the collection and inoculated on MacConkey agar. One hypothesis was that bacteriophages had infected and killed those clinical isolates. To check the best storage conditions to maintain viable P. aeruginosa for a longer time, clinical isolates were stored at various temperatures and were grown monthly. We investigated the presence of phage in 10 clinical isolates of P. aeruginosa stored in our culture collection. Four strains of P. aeruginosa were infected by phages that were characterized by electron microscopy and isolated to assess their ability to infect. The best condition to maintain the viability of the strains during storage was in water at room temperature. Three Siphoviridae and two Myoviridae phages were visualized and characterized by morphology. We confirmed the presence of bacteriophages infecting clinical isolates, and their ability to infect and lyse alternative hosts. Strain PAO1, however, did not show lysis to any phage. Mucoid and multidrug resistant strains of P. aeruginosa showed lysis to 50% of the phages tested.

  14. Simulated effects of dam removal on water temperatures along the Klamath River, Oregon and California, using 2010 Biological Opinion flow requirements

    Science.gov (United States)

    Risley, John C.; Brewer, Scott J.; Perry, Russell W.

    2012-01-01

    Computer model simulations were run to determine the effects of dam removal on water temperatures along the Klamath River, located in south-central Oregon and northern California, using flow requirements defined in the 2010 Biological Opinion of the National Marine Fisheries Service. A one-dimensional, daily averaged water temperature model (River Basin Model-10) developed by the U.S. Environmental Protection Agency Region 10, Seattle, Washington, was used in the analysis. This model had earlier been configured and calibrated for the Klamath River by the U.S. Geological Survey for the U.S. Department of the Interior, Klamath Secretarial Determination to simulate the effects of dam removal on water temperatures for current (2011) and future climate change scenarios. The analysis for this report was performed outside of the scope of the Klamath Secretarial Determination process at the request of the Bureau of Reclamation Technical Services Office, Denver, Colorado.For this analysis, two dam scenarios were simulated: “dams in” and “dams out.” In the “dams in” scenario, existing dams in the Klamath River were kept in place. In the “dams out” scenario, the river was modeled as a natural stream, without the J.C. Boyle, Copco1, Copco2, and Iron Gate Dams, for the entire simulation period. Output from the two dam scenario simulations included daily water temperatures simulated at 29 locations for a 50-year period along the Klamath River between river mile 253 (downstream of Link River Dam) and the Pacific Ocean. Both simulations used identical flow requirements, formulated in the 2010 Biological Opinion, and identical climate conditions based on the period 1961–2009.Simulated water temperatures from January through June at almost all locations between J.C. Boyle Reservoir and the Pacific Ocean were higher for the “dams out” scenario than for the “dams in” scenario. The simulated mean monthly water temperature increase was highest [1.7–2

  15. Aspergillus triggers phenazine production in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Jensen, Britt Guillaume; Jelsbak, Lars; Søndergaard, Ib

    Aspergillus species. Methods: A suspension of fungal spores was streaked onto WATM agar plates. After 24 hours incubation at 37 °C, a P. aeruginosa overnight culture was streaked out perpendicular to the fungal streak. The plates were incubated at 37 °C for five days, examined and plugs were extracted...... for HPLC-DAD and HPLC-DAD-MS analysis. Results: P. aeruginosa PAO1 suppressed growth of A. fumigatus, A. niger, A. flavus, A. oryzae, A. terreus and Emericella nidulans. HPLC and HPLC-DAD-MS results showed an increase in phenazine-1-carboxylic acid and phenazine-1-carboxamide production by P. aeruginosa...

  16. Suppression of Aspergillus by Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Jensen, Britt Guillaume; Jelsbak, Lars; Søndergaard, Ib

    culture plates. After 24 hours incubation at 37 °C, a P. aeruginosa overnight culture diluted to 108 CFU/ml was streaked out perpendicular to the fungal streak. The plates were incubated at 37 °C for 5 days, examined and plugs were extracted for HPLC and LC-DAD-MS analysis. Results: P. aeruginosa PAO1...... suppressed growth of A. fumigatus, A. niger, A. flavus, A. oryzae, A. terreus and E. nidulans. HPLC and LC-DAD-MS results showed an increase in phenazine-1-carboxylic acid and phenazine-1-carboxamide production by P. aeruginosa in the contact area of Aspergillus. Different quinolones were also identified...

  17. Occurrence of Pseudomonas aeruginosa in Kuwait soil.

    Science.gov (United States)

    Al-Saleh, Esmaeil; Akbar, Abrar

    2015-02-01

    Environmentally ubiquitous bacteria such as Pseudomonas aeruginosa evolved mechanisms to adapt and prevail under diverse conditions. In the current investigation, strains of P. aeruginosa demonstrating high rates of crude oil utilization and tolerance to high concentrations of heavy metals were found in both crude oil-contaminated and uncontaminated sites in Kuwait, and were dominant in the contaminated sites. The incidence of P. aeruginosa in tested soils implies the definitive pattern of crude oil contamination in the selection of the bacterial population in petroleum-contaminated sites in Kuwait. Surprisingly, the unculturable P. aeruginosa in different soil samples showed significant high similarity coefficients based on 16S-RFLP analyses, implying that the unculturable fraction of existing bacterial population in environmental samples is more stable and, hence, reliable for phylogenetic studies compared to the culturable bacteria.

  18. Osmoregulation in Pseudomonas aeruginosa under hyperosmotic shock.

    Science.gov (United States)

    Velasco, R; Burgoa, R; Flores, E; Hernández, E; Villa, A; Vaca, S

    1995-01-01

    Pseudomonas aeruginosa PAO1 strain was found to be able to tolerate 700 mM NaCl. 0.5 mM of the osmoprotectant betaine restablished the growth of this strain in 1200 mM NaCl. Intracellular K+ and glutamate concentrations of P. aeruginosa PAO1 after an hyperosmotic shock (400 mM NaCl) showed a permanent increase. Adition of betaine (0.5 mM) to the medium with NaCl had an inhibitory effect on the intracellular accumulation of glutamate. The results indicate that P. aeruginosa PAO1 resists high NaCl concentrations, K+ accumulation and glutamate synthesis probably being the first mechanisms involved in adaptation to osmotic stress. Also is is demonstrated that betaine modulates intracellular glutamate levels in osmotically stressed P. aeruginosa PAO1.

  19. [Clinical features of Pseudomonas aeruginosa infections].

    Science.gov (United States)

    Sarlangue, J; Brissaud, O; Labrèze, C

    2006-10-01

    Pseudomonas aeruginosa is a ubiquitous environmental organism usually considered as opportunistic pathogen in immunocompromised subjects. However it can produce disease in healthy children, mainly on moist body sites. Familial, community and nosocomial outbreaks of cutaneous infections have been reported. Ecthyma gangrenosum is possible without bacteremia. P. aeruginosa is also the most common cause of otitis externa in swimmers and osteomyelitis after puncture wound of the foot.

  20. Dynamics of Pseudomonas aeruginosa genome evolution

    OpenAIRE

    Mathee, Kalai; Narasimhan, Giri; Valdes, Camilo; Qiu, Xiaoyun; Matewish, Jody M.; Koehrsen, Michael; Rokas, Antonis; Yandava, Chandri N.; Engels, Reinhard; Zeng, Erliang; Olavarietta, Raquel; Doud, Melissa; Smith, Roger S.; Montgomery, Philip; White, Jared R.

    2008-01-01

    One of the hallmarks of the Gram-negative bacterium Pseudomonas aeruginosa is its ability to thrive in diverse environments that includes humans with a variety of debilitating diseases or immune deficiencies. Here we report the complete sequence and comparative analysis of the genomes of two representative P. aeruginosa strains isolated from cystic fibrosis (CF) patients whose genetic disorder predisposes them to infections by this pathogen. The comparison of the genomes of the two CF strains...

  1. Interspecific small molecule interactions between clinical isolates of Pseudomonas aeruginosa and Staphylococcus aureus from adult cystic fibrosis patients.

    Directory of Open Access Journals (Sweden)

    Alexandre Fugère

    Full Text Available Pseudomonas aeruginosa and Staphylococcus aureus are the most prevalent pathogens in airway infections of cystic fibrosis (CF patients. We studied how these pathogens coexist and interact with each other. Clinical isolates of both species were retrieved from adult CF patients. Culture supernatants from 63 P. aeruginosa isolates triggered a wide range of biofilm-stimulatory activities when added to the culture of a control S. aureus strain. The extent of biofilm formation by S. aureus was positively correlated to the levels of the 2-alkyl-4-(1H-quinolones (AQs Pseudomonas Quinolone Signal (PQS and 2-heptyl-4-hydroxy quinoline N-oxide (HQNO produced by the P. aeruginosa isolates. Supernatants from P. aeruginosa isogenic mutants deficient in PQS and HQNO production stimulated significantly less biofilm formation by S. aureus than that seen with the parental strain PA14. When studying co-isolated pairs of P. aeruginosa and S. aureus retrieved from patients showing both pathogens, P. aeruginosa supernatants stimulated less biofilm production by the S. aureus counterparts compared to that observed using the control S. aureus strain. Accordingly, some P. aeruginosa isolates produced low levels of exoproducts and also some of the clinical S. aureus isolates were not stimulated by their co-isolates or by PA14 despite adequate production of HQNO. This suggests that colonization of the CF lungs promotes some type of strain selection, or that co-existence requires specific adaptations by either or both pathogens. Results provide insights on bacterial interactions in CF.

  2. Pseudomonas pellicle in disinfectant testing: electron microscopy, pellicle removal, and effect on test results.

    Science.gov (United States)

    Cole, E C; Rutala, W A; Carson, J L; Alfano, E M

    1989-01-01

    Pseudomonas aeruginosa ATCC 15442 is a required organism in the Association of Official Analytical Chemists use-dilution method for disinfectant efficacy testing. When grown in a liquid medium, P. aeruginosa produces a dense mat or pellicle at the broth/air interface. The purpose of this investigation was to examine the pellicle by scanning electron microscopy, to evaluate three pellicle removal methods, and to determine the effect of pellicle fragments on disinfectant efficacy test results. The efficacies of three methods of pellicle removal (decanting, vacuum suction, and filtration) were assessed by quantifying cell numbers on penicylinders. The Association of Official Analytical Chemists use-dilution method was used to determine whether pellicle fragments in the tubes used to inoculate penicylinders affected test results. Scanning electron micrographs showed the pellicle to be a dense mass of intact, interlacing cells at least 10 microns thick. No significant differences in pellicle removal methods were observed, and the presence of pellicle fragments usually increased the number of positive tubes in the use-dilution method significantly. Images PMID:2497711

  3. Non-apoptotic toxicity of Pseudomonas aeruginosa toward murine cells.

    Directory of Open Access Journals (Sweden)

    Sanhita Roy

    Full Text Available Although P. aeruginosa is especially dangerous in cystic fibrosis (CF, there is no consensus as to how it kills representative cell types that are of key importance in the lung. This study concerns the acute toxicity of the sequenced strain, PAO1, toward a murine macrophage cell line (RAW 264.7. Toxicity requires brief contact with the target cell, but is then delayed for more than 12 h. None of the classical toxic effectors of this organism is required and cell death occurs without phagocytosis or acute perturbation of the actin cytoskeleton. Apoptosis is not required for toxicity toward either RAW 264.7 cells or for alveolar macrophages. Transcriptional profiling shows that encounter between PAO1 and RAW 264.7 cells elicits an early inflammatory response, followed by growth arrest. As an independent strategy to understand the mechanism of toxicity, we selected variant RAW 264.7 cells that resist PAO1. Upon exposure to P. aeruginosa, they are hyper-responsive with regard to classical inflammatory cytokine production and show transient downregulation of transcripts that are required for cell growth. They do not show obvious morphologic changes. Although they do not increase interferon transcripts, when exposed to PAO1 they dramatically upregulate a subset of the responses that are characteristic of exposure to g-interferon, including several guanylate-binding proteins. The present observations provide a novel foundation for learning how to equip cells with resistance to a complex challenge.

  4. Responses of Pseudomonas aeruginosa to antimicrobials

    Directory of Open Access Journals (Sweden)

    Yuji eMorita

    2014-01-01

    Full Text Available Infections caused by Pseudomonas aeruginosa often are hard to treat; inappropriate chemotherapy readily selects multidrug-resistant P. aeruginosa. This organism can be exposed to a wide range of concentrations of antimicrobials during treatment; learning more about the responses of P. aeruginosa to antimicrobials is therefore important. We review here responses of the bacterium P. aeruginosa upon exposure to antimicrobials at levels below the inhibitory concentration.Carbapenems (e.g., imipenem have been shown to induce the formation of thicker and more robust biofilms, while fluoroquinolones (e.g., ciprofloxacin and aminoglycosides (e.g., tobramycin have been shown to induce biofilm formation. Ciprofloxacin also has been demonstrated to enhance the frequency of mutation to carbapenem resistance. Conversely, although macrolides (e.g., azithromycin typically are not effective against P. aeruginosa because of the pseudomonal outer-membrane impermeability and efflux, macrolides do lead to a reduction in virulence factor production. Similarly, tetracycline is not very effective against this organism, but is known to induce the type-III secretion system and consequently enhance cytotoxicity of P. aeruginosa in vivo. Of special note are the effects of antibacterials and disinfectants on pseudomonal efflux systems. Sub-inhibitory concentrations of protein synthesis inhibitors (aminoglycosides, tetracycline, chloramphenicol, etc. induce the MexXY multidrug efflux system. This response is known to be mediated by interference with the translation of the leader peptide PA5471.1, with consequent effects on expression of the PA5471 gene product. Additionally, induction of the MexCD-OprJ multidrug efflux system is observed upon exposure to sub-inhibitory concentrations of disinfectants such as chlorhexidine and benzalkonium. This response is known to be dependent upon the AlgU stress response factor.Altogether, these biological responses of P. aeruginosa

  5. Extracellular DNA Acidifies Biofilms and Induces Aminoglycoside Resistance in Pseudomonas aeruginosa.

    Science.gov (United States)

    Wilton, Mike; Charron-Mazenod, Laetitia; Moore, Richard; Lewenza, Shawn

    2015-11-09

    Biofilms consist of surface-adhered bacterial communities encased in an extracellular matrix composed of DNA, exopolysaccharides, and proteins. Extracellular DNA (eDNA) has a structural role in the formation of biofilms, can bind and shield biofilms from aminoglycosides, and induces antimicrobial peptide resistance mechanisms. Here, we provide evidence that eDNA is responsible for the acidification of Pseudomonas aeruginosa planktonic cultures and biofilms. Further, we show that acidic pH and acidification via eDNA constitute a signal that is perceived by P. aeruginosa to induce the expression of genes regulated by the PhoPQ and PmrAB two-component regulatory systems. Planktonic P. aeruginosa cultured in exogenous 0.2% DNA or under acidic conditions demonstrates a 2- to 8-fold increase in aminoglycoside resistance. This resistance phenotype requires the aminoarabinose modification of lipid A and the production of spermidine on the bacterial outer membrane, which likely reduce the entry of aminoglycosides. Interestingly, the additions of the basic amino acid L-arginine and sodium bicarbonate neutralize the pH and restore P. aeruginosa susceptibility to aminoglycosides, even in the presence of eDNA. These data illustrate that the accumulation of eDNA in biofilms and infection sites can acidify the local environment and that acidic pH promotes the P. aeruginosa antibiotic resistance phenotype.

  6. Pseudomoniasis phytotherapy: a review on most important Iranian medicinal plants effective on Pseudomonas aeruginosa

    Science.gov (United States)

    Bahmani, Mahmoud; Rafieian-Kopaei, Mahmoud; Hassanzadazar, Hassan; Taherikalani, Morovat

    2016-01-01

    Background and Objectives: Pseudomonas aeruginosa is a Gram-negative, aerobic bacterium found in water and soil. It is a normal flora in skin and gastrointestinal tract of human beings. P. aeruginosa as an opportunistic pathogen involved in nosocomial infections having multiple pathogenic factors and shows high rate of resistance to different antibiotics. The aim of this study was to identify the most important native medicinal plants of Iran effective on P. aeruginosa. Materials and Methods: All required information was obtained by searching keywords such as P. aeruginosa, medicinal plant extracts or essential oils in published articles in authentic scientific databases such as Science Direct, Wiley-Blackwell, Springer, Google scholar, Scientific Information Database (SID) and Magiran. Results: According to the literature review, our results showed 12 different native medicinal plants were effective against P. aeruginosa in Iran including Eucalyptus camadulensis, Marticaria chamomilla, Ferula gummosa Boiss, Lawsonia inermis, Ocimumgra tissimum, Allium sativum, Satureja hortensis L, Satureja bachtiarica Bunge, Satureja khuzestanica (Jamzad), Thymus daenensis Celak, Thymus carmanicus Jalals and Camellia sinensis. Conclusion: Phytochemical analysis has shown that bioactive compounds of medicinal plants with their antioxidant and antimicrobial properties can be good alternatives for the synthetic medicines in food and drug industry. PMID:28149496

  7. Evaluation of Mannosidase and Trypsin Enzymes Effects on Biofilm Production of Pseudomonas aeruginosa Isolated from Burn Wound Infections

    Science.gov (United States)

    Banar, Maryam; Emaneini, Mohammad; Satarzadeh, Mhboubeh; Abdellahi, Nafiseh; Beigverdi, Reza; van Leeuwen, Willem B.; Jabalameli, Fereshteh

    2016-01-01

    Biofilm is an important virulence factor in Pseudomonas aeruginosa and has a substantial role in antibiotic resistance and chronic burn wound infections. New therapeutic agents against P. aeruginosa, degrading biofilms in burn wounds and improving the efficacy of current antimicrobial agents, are required. In this study, the effects of α-mannosidase, β-mannosidase and trypsin enzymes on the degradation of P. aeruginosa biofilms and on the reduction of ceftazidime minimum biofilm eliminating concentrations (MBEC) were evaluated. All tested enzymes, destroyed the biofilms and reduced the ceftazidime MBECs. However, only trypsin had no cytotoxic effect on A-431 human epidermoid carcinoma cell lines. In conclusion, since trypsin had better features than mannosidase enzymes, it can be a promising agent in combatting P. aeruginosa burn wound infections. PMID:27736961

  8. Outbreak of Pseudomonas aeruginosa bacteraemia in a haematology department

    DEFF Research Database (Denmark)

    Rasmussen, Benjamin Schnack; Christensen, Nikolas; Sørensen, Jan;

    2015-01-01

    INTRODUCTION: Infection by Pseudomonas aeruginosa represents a major cause of morbidity and mortality among immunocompromised patients. In Denmark, an increase in P. aeruginosa isolates from blood cultures from a haematology department prompted a hygienic audit in 2007. METHODS: Blood cultures...

  9. Pseudomonas aeruginosa Dose-Response and Bathing Water Infection

    Science.gov (United States)

    Pseudomonas aeruginosa is the most commonly identified opportunistic pathogen associated with pool acquired bather disease. To better understand why this microorganism poses this protracted problem we recently appraised P. aeruginosa pool risk management. Much is known about the ...

  10. Mechanism of resistance to benzalkonium chloride by Pseudomonas aeruginosa.

    OpenAIRE

    Sakagami, Y; Yokoyama, H; Nishimura, H.; Ose, Y; Tashima, T.

    1989-01-01

    The mechanisms of resistance of Pseudomonas aeruginosa to benzalkonium chloride (BC) were studied. The effluence of cell components was observed in susceptible P. aeruginosa by electron microscopy, but resistant P. aeruginosa seemed to be undamaged. No marked changes in cell surface potential between Escherichia coli NIHJC-2 and a spheroplast strain were found. The contents of phospholipids (PL) and fatty and neutral lipids (FNL) in the cell walls of resistant P. aeruginosa were higher than t...

  11. Vaccines for preventing infection with Pseudomonas aeruginosa in cystic fibrosis

    DEFF Research Database (Denmark)

    Johansen, Helle Krogh; Gøtzsche, Peter C

    2013-01-01

    Chronic pulmonary infection in cystic fibrosis results in progressive lung damage. Once colonisation of the lungs with Pseudomonas aeruginosa occurs, it is almost impossible to eradicate. Vaccines, aimed at reducing infection with Pseudomonas aeruginosa, have been developed.......Chronic pulmonary infection in cystic fibrosis results in progressive lung damage. Once colonisation of the lungs with Pseudomonas aeruginosa occurs, it is almost impossible to eradicate. Vaccines, aimed at reducing infection with Pseudomonas aeruginosa, have been developed....

  12. Effects of two copper compounds on Microcystis aeruginosa cell density, membrane integrity, and microcystin release.

    Science.gov (United States)

    Tsai, Kuo-Pei

    2015-10-01

    Microcystin release following Microcystis aeruginosa cell lysis after copper-based algaecide treatment is often cited as a concern leading to restricted use of algaecide in restoration of natural water resources. To examine this concern, bench-scale experiments were conducted to study responses of M. aeruginosa to 8-day copper exposures as copper sulfate and copper-ethanolamine (Cu-EA). M. aeruginosa UTEX 2385 was cultured in BG11 medium to cell density of 10(6)cells/mL with total and extracellular microcystin of 93 and 53μg/L, respectively. Exposures of copper concentration ranged from 40 to 1000μgCu/L. Cell membrane integrity was indicated by erythrosine B. In the end of experiment, total microcystin and cell density in untreated control (313μg/L and 10(7)cells/mL) was 3.3 and 10 times greater than pretreatment value, respectively. Minimum amount of copper required to reduce M. aeruginosa population within 8 days was 160μgCu/L as copper sulfate and 80μgCu/L as Cu-EA, where total and extracellular microcystin concentrations (47 and 44μg/L for copper sulfate; 56 and 44μg/L for Cu-EA) were degraded with degradation rate coefficient 0.1 day(-1) and were less than pretreatment values. Given a copper concentration at 80µgCu/L as Cu-EA, M. aeruginosa cells were intact and less microcystin were released compared to treatments at 160-1000µgCu/L, where lysed cells and relatively greater microcystin release were observed. Based on the laboratory results, a minimum amount of copper required for reducing M. aeruginosa population could decrease total microcystin concentration and not compromise cells and minimize microcystin release.

  13. Glycopeptide dendrimers as Pseudomonas aeruginosa biofilm inhibitors.

    Science.gov (United States)

    Reymond, Jean-Louis; Bergmann, Myriam; Darbre, Tamis

    2013-06-01

    Synthetic glycopeptide dendrimers composed of a branched oligopeptide tree structure appended with glycosidic groups at its multiple N-termini were investigated for binding to the Pseudomonas aeruginosa lectins LecB and LecA. These lectins are partly responsible for the formation of antibiotic resistant biofilms in the human pathogenic bacterium P. aeruginosa, which causes lethal airway infections in immune-compromised and cystic fibrosis patients. Glycopeptide dendrimers with high affinity to the lectins were identified by screening of combinatorial libraries. Several of these dendrimers, in particular the LecB specific glycopeptide dendrimers FD2 and D-FD2 and the LecA specific glycopeptide dendrimers GalAG2 and GalBG2, also efficiently block P. aeruginosa biofilm formation and induce biofilm dispersal in vitro. Structure-activity relationship and structural studies are reviewed, in particular the observation that multivalency is essential to the anti-biofilm effect in these dendrimers.

  14. Complement activation by Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Jensen, E T; Kharazmi, A; Garred, P

    1993-01-01

    In chronic infections, such as the bronchopulmonary Pseudomonas aeruginosa infection in cystic fibrosis (CF) patients, bacteria persist despite an intact host immune defense and frequent antibiotic treatment. An important reason for the persistence of the bacteria is their capacity for the biofilm...... immuno-electrophoresis, C5a generation tested by a PMN chemotactic assay, and terminal complement complex formation measured by ELISA. Two of the four assays showed that P. aeruginosa grown in biofilm activated complement less than planktonic bacteria, and all assays showed that activation by intact...... influx of neutrophils are known to cause inflammatory changes in the lungs. P. aeruginosa persisting in biofilms may contribute to the constant inflammation taking place in the lungs of CF patients....

  15. Stratified growth in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Werner, E.; Roe, F.; Bugnicourt, A.;

    2004-01-01

    In this study, stratified patterns of protein synthesis and growth were demonstrated in Pseudomonas aeruginosa biofilms. Spatial patterns of protein synthetic activity inside biofilms were characterized by the use of two green fluorescent protein (GFP) reporter gene constructs. One construct...... carried an isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible gfpmut2 gene encoding a stable GFP. The second construct carried a GFP derivative, gfp-AGA, encoding an unstable GFP under the control of the growth-rate-dependent rrnBp(1) promoter. Both GFP reporters indicated that active protein...... of oxygen limitation in the biofilm. Oxygen microelectrode measurements showed that oxygen only penetrated approximately 50 mum into the biofilm. P. aeruginosa was incapable of anaerobic growth in the medium used for this investigation. These results show that while mature P. aeruginosa biofilms contain...

  16. Hair Removal

    Science.gov (United States)

    ... Loss Surgery? A Week of Healthy Breakfasts Shyness Hair Removal KidsHealth > For Teens > Hair Removal A A A ... recommend an electrologist with the proper credentials. Laser Hair Removal How It Works: A laser is directed through ...

  17. Cyanobacterium Microcystis aeruginosa response to pentachlorophenol and comparison with that of the microalga Chlorella vulgaris.

    Science.gov (United States)

    de Morais, Paulo; Stoichev, Teodor; Basto, M Clara P; Ramos, V; Vasconcelos, V M; Vasconcelos, M Teresa S D

    2014-04-01

    Pentachlorophenol (PCP) effects on a strain of the cyanobacterium Microcystis aeruginosa were investigated at laboratory scale. This is the first systematic ecotoxicity study of the effects of PCP on an aquatic cyanobacterium. The microalga Chlorella vulgaris was studied in the same conditions as the cyanobacterium, in order to compare the PCP toxicity and its removal by the species. The cells were exposed to environmental levels of PCP during 10 days, in Fraquil culture medium, at nominal concentrations from 0.01 to 1000 μg L(-1), to the cyanobacterium, and 0.01 to 5000 μg L(-1), to the microalga. Growth was assessed by area under growth curve (AUC, optical density vs time) and chlorophyll a content (chla). The toxicity profiles of the two species were very different. The calculated effective concentrations EC20 and EC50 were much lower to M. aeruginosa, and its growth inhibition expressed by chla was concentration-dependent while by AUC was not concentration-dependent. The cells might continue to divide even with lower levels of chla. The number of C. vulgaris cells decreased with the PCP concentration without major impact on the chla. The effect of PCP on M. aeruginosa is hormetic: every concentration studied was toxic except 1 μg L(-1), which promoted its growth. The legal limit of PCP set by the European Union for surface waters (1 μg L(-1)) should be reconsidered since a toxic cyanobacteria bloom might occur. The study of the removal of PCP from the culture medium by the two species is an additional novelty of this work. M. aeruginosa could remove part of the PCP from the medium, at concentrations where toxic effects were observed, while C. vulgaris stabilized it.

  18. QapR (PA5506) represses an operon that negatively affects the Pseudomonas quinolone signal in Pseudomonas aeruginosa.

    Science.gov (United States)

    Tipton, Kyle A; Coleman, James P; Pesci, Everett C

    2013-08-01

    Pseudomonas aeruginosa is a Gram-negative, opportunistic pathogen that can cause disease in varied sites within the human body and is a significant source of morbidity and mortality in those afflicted with cystic fibrosis. P. aeruginosa is able to coordinate group behaviors, such as virulence factor production, through the process of cell-to-cell signaling. There are three intercellular signaling systems employed by P. aeruginosa, and one of these systems utilizes the small molecule 2-heptyl-3-hydroxy-4-quinolone (Pseudomonas quinolone signal [PQS]). PQS is required for virulence in multiple infection models and has been found in the lungs of cystic fibrosis patients colonized by P. aeruginosa. In this study, we have identified an RpiR family transcriptional regulator, QapR, which is an autoregulatory repressor. We found that mutation of qapR caused overexpression of the qapR operon. We characterized the qapR operon to show that it contains genes qapR, PA5507, PA5508, and PA5509 and that QapR directly controls the transcription of these genes in a negative manner. We also show that derepression of this operon greatly reduces PQS concentration in P. aeruginosa. Our results suggest that qapR affects PQS concentration by repressing an enzymatic pathway that acts on PQS or a PQS precursor to lower the PQS concentration. We believe that this operon comprises a novel mechanism to regulate PQS concentration in P. aeruginosa.

  19. Control of Pseudomonas aeruginosa and Stenotrophomonas maltophilia contamination of microfiltered water dispensers with peracetic acid and hydrogen peroxide.

    Science.gov (United States)

    Sacchetti, Rossella; De Luca, Giovanna; Zanetti, Franca

    2009-06-30

    The abilities of peracetic acid and hydrogen peroxide to remove or reduce Pseudomonas aeruginosa and Stenotrophomonas maltophilia in output water from microfiltered water dispensers (MWDs) were investigated. Two MWDs were inoculated with strains of P. aeruginosa and S. maltophilia isolated from water. Dispensers A and B were disinfected with 10% (v/v) peracetic acid (PAA) and 3% (v/v) hydrogen peroxide (HP) respectively. Each dispenser was disinfected three times at monthly intervals with contact times of 10, 30 and 40 min. Water dispensed by the MWDs was collected immediately before and after each treatment and then twice weekly for the remaining period. Once a week a sample of the tap water entering the dispensers was tested. P. aeruginosa and S. maltophilia were enumerated in the 90 samples collected during 6 months. In the output water from the dispensers before the first treatment, the number of the bacteria was 3 to 4 log cfu/100 mL. Treatment with PAA greatly reduced the numbers of P. aeruginosa and S. maltophilia in the dispensed water initially. However, by 2 days after treatment, the numbers increased and remained high. In the case of disinfection with HP for 40 min, P. aeruginosa was not detected in most of the samples (73.7%). Numbers of S. maltophilia decreased with increasing time after treatment.

  20. Standardized chemical synthesis of Pseudomonas aeruginosa pyocyanin

    Directory of Open Access Journals (Sweden)

    Rajkumar Cheluvappa

    2014-01-01

    As we have extracted pyocyanin both from P. aeruginosa cultures, and via chemical synthesis; we know the procedural and product-quality differences. We endorse the relative ease, safety, and convenience of using the chemical synthesis described here. Crucially, our “naturally endotoxin-free” pyocyanin can be extracted easily without using infectious bacteria.

  1. Phospholipase C from Pseudomonas aeruginosa and Bacillus cereus;characterization of catalytic activity

    Institute of Scientific and Technical Information of China (English)

    Nooran Sherif Elleboudy; Mohammad Mabrouk Aboulwafa; Nadia Abdel-Haleem Hassouna

    2014-01-01

    Objective:To study characteristics of phospholipases C (PLCs), their importance for producing microorganisms as well as the potential of their use for industrial purposes. Methods:PLC from Bacillus cereus (B. cereus) D101 was selected as an example of Gram-positive PLCs and PLC from Pseudomonas aeruginosa (P. aeruginosa) D183 of Gram-negative ones. Enzymes were partially purified by ammonium sulfate precipitation followed by membrane dialysis. Partially purified preparations were used to study effect of different factors on activities as well as in substrate specificity tests which were conducted using a turbidimetric assay method. Results: Maximum activity was at pH 7 and 8 and 40℃for P. aeruginosa PLC, and pH 8-10 and 37℃for B. cereus PLC. Both PLCs were inhibited by Pi at 5 mM or higher, whereas, PLC from B. cereus only was inhibited by EDTA. Activity of P. aeruginosa PLC was not affected by removing Zn2+ions from reaction mixture or their replacement with Ca2+, Ba2+, Mg2+or Mn2+ions. Vis-à-vis, activity of B. cereus PLC was found to be metal ion dependent. PLCs from both isolates were relatively thermostable and showed maximum affinity toward phosphatidylcholine. Sphingomyelin and phosphatidylethanolamine were not good substrates and phosphatidylinositol, phosphatidylserine, phosphatidylglycerol and cardiolipin could be considered non-substrates. Conclusions: Human body physiological conditions could favor activity of P. aeruginosa and B. cereus PLCs. These enzymes may participate in phosphate scavenging and virulence of producing isolates but not in autolysis. PLCs from both isolates are potential candidates for industrial use.

  2. Pseudomonas aeruginosa SoxR does not conform to the archetypal paradigm for SoxR-dependent regulation of the bacterial oxidative stress adaptive response.

    Science.gov (United States)

    Palma, Marco; Zurita, Juan; Ferreras, Julian A; Worgall, Stefan; Larone, Davise H; Shi, Lei; Campagne, Fabien; Quadri, Luis E N

    2005-05-01

    SoxR is a transcriptional regulator that controls an oxidative stress response in Escherichia coli. The regulator is primarily activated by superoxide anion-dependent oxidation. Activated SoxR turns on transcription of a single gene, soxS, which encodes a transcriptional regulator that activates a regulon that includes dozens of oxidative stress response genes. SoxR homologues have been identified in many bacterial species, including the opportunistic pathogen Pseudomonas aeruginosa. However, the expected SoxR partner, SoxS, has not been found in P. aeruginosa. Thus, the primary gene target(s) of P. aeruginosa SoxR is unknown and the involvement of this regulator in the oxidative stress response of the bacterium remains unclear. We utilized transcriptome profiling to identify the P. aeruginosa SoxR regulon and constructed and characterized an unmarked P. aeruginosa DeltasoxR mutant. We provide evidence indicating that P. aeruginosa SoxR activates a six-gene regulon in response to O(2)(.-)-induced stress. The regulon includes three transcriptional units: (i) the recently identified mexGHI-ompD four-gene operon, which encodes a multidrug efflux pump system involved in quorum-sensing signal homeostasis; (ii) gene PA3718, encoding a probable efflux pump; and (iii) gene PA2274, encoding a probable monooxygenase. We also demonstrate that P. aeruginosa SoxR is not a key regulatory player in the oxidative stress response. Finally, we show that P. aeruginosa SoxR is required for virulence in a mouse model of intrapulmonary infection. These results demonstrate that the E. coli-based SoxRS paradigm does not hold in P. aeruginosa and foster new hypotheses for the possible physiological role of P. aeruginosa SoxR.

  3. The behaviors of Microcystis aeruginosa cells and extracellular microcystins during chitosan flocculation and flocs storage processes.

    Science.gov (United States)

    Pei, Hai-Yan; Ma, Chun-Xia; Hu, Wen-Rong; Sun, Feng

    2014-01-01

    This work aimed to study the effects of chitosan on cell integrity and extracellular microcystins (MCs) of Microcystis aeruginosa cells during flocculation and flocs storage processes. The impacts of chitosan addition, flocculation stirring and flocs storage time were comprehensively detected to prevent or reduce cell lysis and MCs release. Response surface method (RSM) was applied to optimize the chitosan flocculation. Under chitosan concentration 7.31 mg/L and optimized mechanical conditions, 99% of M. aeruginosa cells were integrated removed. Furthermore, amounts of extracellular MCs were adsorbed by chitosan polymers in this process. With chitosan flocs protect, though cells showed some damage, extracellular MCs concentration in flocculated samples lower than background level within first 2 d. However, lots of MCs release was observed after 4d which may result from chitosan degradation and cells lysis. Therefore, chitosan flocs should be treated within 2d to prevent the adsorbed MCs releasing again.

  4. phoU inactivation in Pseudomonas aeruginosa enhances accumulation of ppGpp and polyphosphate.

    Science.gov (United States)

    de Almeida, Luiz Gustavo; Ortiz, Julia Helena; Schneider, René P; Spira, Beny

    2015-05-01

    Inorganic polyphosphate (polyP) is a linear polymer composed of several molecules of orthophosphate (Pi) linked by energy-rich phosphoanhydride bonds. In Pseudomonas aeruginosa, Pi is taken up by the ABC transporter Pst, encoded by an operon consisting of five genes. The first four genes encode proteins involved in the transport of Pi and the last gene of the operon, phoU, codes for a protein which exact function is unknown. We show here that the inactivation of phoU in P. aeruginosa enhanced Pi removal from the medium and polyP accumulation. The phoU mutant also accumulated high levels of the alarmone guanosine tetraphosphate (ppGpp), which in turn increased the buildup of polyP. In addition, phoU inactivation had several pleiotropic effects, such as reduced growth rate and yield and increased sensitivity to antibiotics and stresses. However, biofilm formation was not affected by the phoU mutation.

  5. Comparison of quantification methods illustrates reduced Pseudomonas aeruginosa activity on nanorough polyvinyl chloride.

    Science.gov (United States)

    Seil, Justin T; Rubien, Nathan M; Webster, Thomas J; Tarquinio, Keiko M

    2011-07-01

    Patients on mechanical ventilators for extended periods of time are faced with a high probability of developing ventilator associated pneumonia. Although this has been mostly addressed through the re-engineering of endotracheal tubes (ETTs) with antimicrobial materials, such material coatings may easily delaminate during use. However, the potential exists to apply nanotechnology to the ETT to avoid delamination but implement antibacterial properties. Selecting a protocol to evaluate in vitro material for anti-infection is difficult, partially due to the existence of conflicting reported methods of analysis. In this study, the susceptibility of conventional and nanorough polymeric materials to bacterial biofilm growth were evaluated. After creating nanorough polyvinyl chloride (PVC) ETTs, Pseudomonas aeruginosa biofilms were then grown on sample surfaces during a 24-h culture. Biofilms were then removed and assayed from sample surfaces using a variety of techniques. Comparisons between the different techniques used for biofilm removal indicated that vortexing provided adequate removal of the biofilm from sample surfaces. Most importantly, a protocol following the vortexing method of biofilm and bacteria removal provided an ∼40% lower yield of colony forming units from nanorough PVC compared to conventional PVC. This suggests that Pseudomonas aeruginosa are less adherent on nanorough PVC than conventional PVC.

  6. Tick Removal

    Science.gov (United States)

    ... ticks Tickborne diseases abroad Borrelia miyamotoi Borrelia mayonii Tick Removal Recommend on Facebook Tweet Share Compartir If ... a tick quite effectively. How to remove a tick Use fine-tipped tweezers to grasp the tick ...

  7. Cadmium removal from contaminated soil by tunable biopolymers.

    Science.gov (United States)

    Prabhukumar, Giridhar; Matsumoto, Mark; Mulchandani, Ashok; Chen, Wilfred

    2004-06-01

    An elastin-like polypeptide (ELP) composed of a polyhistidine tail (ELPH12) was exploited as a tunable, metal-binding biopolymer with high affinity toward cadmium. By taking advantage of the property of ELPH12 to undergo a reversible thermal precipitation, easy recovery of the sequestered cadmium from contaminated water was demonstrated as the result of a simple temperature change. In this study, batch soil washing experiments were performed to evaluate the feasibility of using ELPH12 as an environmentally benign strategy for removing cadmium from contaminated soil. The stability constant (log KL) for the cadmium-ELPH12 complex was determined to be 6.8, a value similar to that reported for the biosurfactant rhamnolipid. Two washings with 1.25 mg/mL of ELPH12 were able to remove more than 55% of the bound cadmium as compared to only 8% removal with ELP containing no histidine tail or 21% removal using the same concentration of EDTA. Unlike rhamnolipid from Pseudomonas aeruginosa ATCC 9027, which adsorbs extensively to soil, less than 10% of ELPH12 was adsorbed under all soil washing conditions. As a result, a significantly lower concentration of ELPH12 (0.036 mM as compared to 5-10 mM of biosurfactants) was required to achieve similar extraction efficiencies. However, cadmium recovery by simple precipitation was incomplete due to the displacement of bound cadmium by zinc ions present in soil. Owing to its benign nature, ease of production, and selective tailoring of the metal binding domain toward any target metals of interest, ELP biopolymers may find utility as an effective extractant for heavy metal removal from contaminated soil or ore processing.

  8. Plate removal following orthognathic surgery.

    Science.gov (United States)

    Little, Mhairi; Langford, Richard Julian; Bhanji, Adam; Farr, David

    2015-11-01

    The objectives of this study are to determine the removal rates of orthognathic plates used during orthognathic surgery at James Cook University Hospital and describe the reasons for plate removal. 202 consecutive orthognathic cases were identified between July 2004 and July 2012. Demographics and procedure details were collected for these patients. Patients from this group who returned to theatre for plate removal between July 2004 and November 2012 were identified and their notes were analysed for data including reason for plate removal, age, smoking status, sex and time to plate removal. 3.2% of plates were removed with proportionally more plates removed from the mandible than the maxilla. 10.4% of patients required removal of one or more plate. Most plates were removed within the first post-operative year. The commonest reasons for plate removal were plate exposure and infection. The plate removal rates in our study are comparable to those seen in the literature.

  9. Biological cost of pyocin production during the SOS response in Pseudomonas aeruginosa.

    Science.gov (United States)

    Penterman, Jon; Singh, Pradeep K; Walker, Graham C

    2014-09-01

    LexA and two structurally related regulators, PrtR and PA0906, coordinate the Pseudomonas aeruginosa SOS response. RecA-mediated autocleavage of LexA induces the expression of a protective set of genes that increase DNA damage repair and tolerance. In contrast, RecA-mediated autocleavage of PrtR induces antimicrobial pyocin production and a program that lyses cells to release the newly synthesized pyocin. Recently, PrtR-regulated genes were shown to sensitize P. aeruginosa to quinolones, antibiotics that elicit a strong SOS response. Here, we investigated the mechanisms by which PrtR-regulated genes determine antimicrobial resistance and genotoxic stress survival. We found that induction of PrtR-regulated genes lowers resistance to clinically important antibiotics and impairs the survival of bacteria exposed to one of several genotoxic agents. Two distinct mechanisms mediated these effects. Cell lysis genes that are induced following PrtR autocleavage reduced resistance to bactericidal levels of ciprofloxacin, and production of extracellular R2 pyocin was lethal to cells that initially survived UV light treatment. Although typically resistant to R2 pyocin, P. aeruginosa becomes transiently sensitive to R2 pyocin following UV light treatment, likely because of the strong downregulation of lipopolysaccharide synthesis genes that are required for resistance to R2 pyocin. Our results demonstrate that pyocin production during the P. aeruginosa SOS response carries both expected and unexpected costs.

  10. Pseudomonas aeruginosa endophthalmitis masquerading as chronic uveitis

    Directory of Open Access Journals (Sweden)

    Kalpana Badami Nagaraj

    2013-01-01

    Full Text Available A 65-year-old male presented with decreased vision in the left eye of 15-day duration after having undergone an uneventful cataract surgery 10 months back. He had been previously treated with systemic steroids for recurrent uveitis postoperatively on three occasions in the same eye. B-scan ultrasonography showed multiple clumplike echoes suggestive of vitreous inflammation. Aqueous tap revealed Pseudomonas aeruginosa sensitive to ciprofloxacin. The patient was treated with intravitreal ciprofloxacin and vancomycin along with systemic ciprofloxacin with good clinical response. Even a virulent organism such as P.aeruginosa can present as a chronic uveitis, which, if missed, can lead to a delay in accurate diagnosis and appropriate management.

  11. Pseudomonas aeruginosa biofilms in cystic fibrosis

    DEFF Research Database (Denmark)

    Høiby, Niels; Ciofu, Oana; Bjarnsholt, Thomas

    2010-01-01

    The persistence of chronic Pseudomonas aeruginosa lung infections in cystic fibrosis (CF) patients is due to biofilm-growing mucoid (alginate-producing) strains. A biofilm is a structured consortium of bacteria, embedded in a self-produced polymer matrix consisting of polysaccharide, protein...... and DNA. In CF lungs, the polysaccharide alginate is the major part of the P. aeruginosa biofilm matrix. Bacterial biofilms cause chronic infections because they show increased tolerance to antibiotics and resist phagocytosis, as well as other components of the innate and the adaptive immune system....... As a consequence, a pronounced antibody response develops, leading to immune complex-mediated chronic inflammation, dominated by polymorphonuclear leukocytes. The chronic inflammation is the major cause of the lung tissue damage in CF. Biofilm growth in CF lungs is associated with an increased frequency...

  12. Cell death in Pseudomonas aeruginosa biofilm development

    DEFF Research Database (Denmark)

    Webb, J.S.; Thompson, L.S.; James, S.

    2003-01-01

    . However, key developmental processes regulating these events are poorly understood. A normal component of multicellular development is cell death. Here we report that a repeatable pattern of cell death and lysis occurs in biofilms of P. aeruginosa during the normal course of development. Cell death....... We propose that prophage-mediated cell death is an important mechanism of differentiation inside microcolonies that facilitates dispersal of a subpopulation of surviving cells....

  13. Pseudomonas aeruginosa genomic structure and diversity

    Directory of Open Access Journals (Sweden)

    Jens eKlockgether

    2011-07-01

    Full Text Available The Pseudomonas aeruginosa genome (G + C content 65-67%, size 5.5 – 7 Mbp is made up of a single circular chromosome and a variable number of plasmids. Sequencing of complete genomes or blocks of the accessory genome has revealed that the genome encodes a large repertoire of transporters, transcriptional regulators and two-component regulatory systems which reflects its metabolic diversity to utilize a broad range of nutrients. The conserved core component of the genome is largely collinear among P. aeruginosa strains and exhibits an interclonal sequence diversity of 0.5 – 0.7%. Only a few loci of the core genome are subject to diversifying selection. Genome diversity is mainly caused by accessory DNA elements located in 79 regions of genome plasticity that are scattered around the genome and show an anomalous usage of mono- to tetradecanucleotides. Genomic islands of the pKLC102/PAGI-2 family that integrate into tRNALys or tRNAGly genes represent hotspots of inter- and intraclonal genomic diversity. The individual islands differ in their repertoire of metabolic genes that make a large contribution to the pangenome. In order to unravel intraclonal diversity of P. aeruginosa, the genomes of two members of the PA14 clonal complex from diverse habitats and geographic origin were compared. The genome sequences differed by less than 0.01% from each other. 198 of the 231 SNPs were non-randomly distributed in the genome. Non-synonymous SNPs were mainly found in an integrated Pf1-like phage and in genes involved in transcriptional regulation, membrane and extracellular constituents, transport and secretion. In summary, P. aeruginosa is endowed with a highly conserved core genome of low sequence diversity and a highly variable accessory genome that communicates with other pseudomonads and genera via horizontal gene transfer.

  14. Nosocomial infections due to Pseudomonas aeruginosa: review of recent trends.

    Science.gov (United States)

    Cross, A; Allen, J R; Burke, J; Ducel, G; Harris, A; John, J; Johnson, D; Lew, M; MacMillan, B; Meers, P

    1983-01-01

    The role of Pseudomonas aeruginosa in nosocomial infections occurring since 1975 is reviewed. Data from the National Nosocomial Infections Study conducted by the Centers for Disease Control, from individual medical centers, and from the literature were used to compare the relative frequency of occurrence of nosocomial infection caused by P. aeruginosa with that of infection caused by other gram-negative bacilli. The relative frequency of P. aeruginosa as a nosocomial pathogen has increased, although wide variations are seen among individual medical centers. P. aeruginosa continues to be a major pathogen among patients with immunosuppression, cystic fibrosis, malignancy, and trauma. While Staphylococcus aureus has become the predominant pathogen in some large burn centers, P. aeruginosa is the most important gram-negative pathogen. Periodic review of the epidemiology of P. aeruginosa infection is warranted in view of the changing incidence of infection caused by this organism.

  15. Glycerol metabolism promotes biofilm formation by Pseudomonas aeruginosa.

    Science.gov (United States)

    Scoffield, Jessica; Silo-Suh, Laura

    2016-08-01

    Pseudomonas aeruginosa causes persistent infections in the airways of cystic fibrosis (CF) patients. Airway sputum contains various host-derived nutrients that can be utilized by P. aeruginosa, including phosphotidylcholine, a major component of host cell membranes. Phosphotidylcholine can be degraded by P. aeruginosa to glycerol and fatty acids to increase the availability of glycerol in the CF lung. In this study, we explored the role that glycerol metabolism plays in biofilm formation by P. aeruginosa. We report that glycerol metabolism promotes biofilm formation by both a chronic CF isolate (FRD1) and a wound isolate (PAO1) of P. aeruginosa. Moreover, loss of the GlpR regulator, which represses the expression of genes involved in glycerol metabolism, enhances biofilm formation in FRD1 through the upregulation of Pel polysaccharide. Taken together, our results suggest that glycerol metabolism may be a key factor that contributes to P. aeruginosa persistence by promoting biofilm formation.

  16. Antivirulence activity of azithromycin in Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Francesco eImperi

    2014-04-01

    Full Text Available Antibiotics represent our bulwark to combat bacterial infections, but the spread of antibiotic resistance compromises their clinical efficacy. Alternatives to conventional antibiotics are urgently needed in order to complement the existing antibacterial arsenal. The macrolide antibiotic azithromycin (AZM provides a paradigmatic example of an unconventional antibacterial drug. Besides its growth-inhibiting activity, AZM displays potent anti-inflammatory properties, as well as antivirulence activity on some intrinsically resistant bacteria, such as Pseudomonas aeruginosa. In this bacterium, the antivirulence activity of AZM mainly relies on its ability to interact with the ribosome, resulting in direct and/or indirect repression of specific subsets of genes involved in virulence, quorum sensing, biofilm formation and intrinsic antibiotic resistance. Both clinical experience and clinical trials have shown the efficacy of AZM in the treatment of chronic pulmonary infections caused by P. aeruginosa. The aim of this review is to combine results from laboratory studies with evidence from clinical trials in order to unify the information on the in vivo mode of action of AZM in P. aeruginosa infection.

  17. Removal of broken hardware.

    Science.gov (United States)

    Hak, David J; McElvany, Matthew

    2008-02-01

    Despite advances in metallurgy, fatigue failure of hardware is common when a fracture fails to heal. Revision procedures can be difficult, usually requiring removal of intact or broken hardware. Several different methods may need to be attempted to successfully remove intact or broken hardware. Broken intramedullary nail cross-locking screws may be advanced out by impacting with a Steinmann pin. Broken open-section (Küntscher type) intramedullary nails may be removed using a hook. Closed-section cannulated intramedullary nails require additional techniques, such as the use of guidewires or commercially available extraction tools. Removal of broken solid nails requires use of a commercial ratchet grip extractor or a bone window to directly impact the broken segment. Screw extractors, trephines, and extraction bolts are useful for removing stripped or broken screws. Cold-welded screws and plates can complicate removal of locked implants and require the use of carbide drills or high-speed metal cutting tools. Hardware removal can be a time-consuming process, and no single technique is uniformly successful.

  18. Pseudomonas aeruginosa Alginate Overproduction Promotes Coexistence with Staphylococcus aureus in a Model of Cystic Fibrosis Respiratory Infection

    Science.gov (United States)

    Limoli, Dominique H.; Whitfield, Gregory B.; Kitao, Tomoe; Ivey, Melissa L.; Davis, Michael R.; Grahl, Nora; Hogan, Deborah A.; Rahme, Laurence G.; Howell, P. Lynne

    2017-01-01

    ABSTRACT While complex intra- and interspecies microbial community dynamics are apparent during chronic infections and likely alter patient health outcomes, our understanding of these interactions is currently limited. For example, Pseudomonas aeruginosa and Staphylococcus aureus are often found to coinfect the lungs of patients with cystic fibrosis (CF), yet these organisms compete under laboratory conditions. Recent observations that coinfection correlates with decreased health outcomes necessitate we develop a greater understanding of these interbacterial interactions. In this study, we tested the hypothesis that P. aeruginosa and/or S. aureus adopts phenotypes that allow coexistence during infection. We compared competitive interactions of P. aeruginosa and S. aureus isolates from mono- or coinfected CF patients employing in vitro coculture models. P. aeruginosa isolates from monoinfected patients were more competitive toward S. aureus than P. aeruginosa isolates from coinfected patients. We also observed that the least competitive P. aeruginosa isolates possessed a mucoid phenotype. Mucoidy occurs upon constitutive activation of the sigma factor AlgT/U, which regulates synthesis of the polysaccharide alginate and dozens of other secreted factors, including some previously described to kill S. aureus. Here, we show that production of alginate in mucoid strains is sufficient to inhibit anti-S. aureus activity independent of activation of the AlgT regulon. Alginate reduces production of siderophores, 2-heptyl-4-hydroxyquinolone-N-oxide (HQNO), and rhamnolipids—each required for efficient killing of S. aureus. These studies demonstrate alginate overproduction may be an important factor driving P. aeruginosa coinfection with S. aureus. PMID:28325763

  19. Crystal structure of PvdO from Pseudomonas aeruginosa.

    Science.gov (United States)

    Yuan, Zenglin; Gao, Fei; Bai, Guohui; Xia, Hengchuan; Gu, Lichuan; Xu, Sujuan

    2017-02-26

    Pyoverdine I (PVDI) is a water-soluble fluorescein siderophore with strong iron chelating ability from the gram-negative pathogen Pseudomonas aeruginosa PAO1. Compared to common siderophores, PVDI is a relatively large compound whose synthesis requires a group of enzymes with different catalytic activities. In addition to four nonribosomal peptide synthetases (NRPS) which are responsible for the production of the peptide backbone of PVDI, several additional enzymes are associated with the modification of the side chains. PvdO is one of these enzymes and participates in PVDI precursor maturation in the periplasm. We determined the crystal structure of PvdO at 1.24 Å resolution. The PvdO structure shares a common fold with some FGly-generating enzymes (FGE) and is stabilized by Ca(2+). However, the catalytic residues in FGE are not observed in PvdO, indicating PvdO adopts a unique catalytic mechanism.

  20. Mechanism of resistance to benzalkonium chloride by Pseudomonas aeruginosa.

    Science.gov (United States)

    Sakagami, Y; Yokoyama, H; Nishimura, H; Ose, Y; Tashima, T

    1989-01-01

    The mechanisms of resistance of Pseudomonas aeruginosa to benzalkonium chloride (BC) were studied. The effluence of cell components was observed in susceptible P. aeruginosa by electron microscopy, but resistant P. aeruginosa seemed to be undamaged. No marked changes in cell surface potential between Escherichia coli NIHJC-2 and a spheroplast strain were found. The contents of phospholipids (PL) and fatty and neutral lipids (FNL) in the cell walls of resistant P. aeruginosa were higher than those in the cell walls of susceptible P. aeruginosa. The amounts of BC adsorbed to PL and FNL of cell walls of BC-resistant P. aeruginosa were lower than those for BC-susceptible P. aeruginosa. Fifteen species of cellular fatty acids were identified by capillary gas chromatography and gas chromatography-mass spectrometry. The ability of BC to permeate the cell wall was reduced because of the increase in cellular fatty acids. These results suggested that the resistance of P. aeruginosa to BC is mainly a result of increased in the contents of PL and FNL. In resistant P. aeruginosa, the decrease in the amount of BC adsorbed is likely to be the result of increases in the contents of PL and FNL. Images PMID:2506813

  1. Pseudomonas aeruginosa Alginate Overproduction Promotes Coexistence with Staphylococcus aureus in a Model of Cystic Fibrosis Respiratory Infection.

    Science.gov (United States)

    Limoli, Dominique H; Whitfield, Gregory B; Kitao, Tomoe; Ivey, Melissa L; Davis, Michael R; Grahl, Nora; Hogan, Deborah A; Rahme, Laurence G; Howell, P Lynne; O'Toole, George A; Goldberg, Joanna B

    2017-03-21

    While complex intra- and interspecies microbial community dynamics are apparent during chronic infections and likely alter patient health outcomes, our understanding of these interactions is currently limited. For example, Pseudomonas aeruginosa and Staphylococcus aureus are often found to coinfect the lungs of patients with cystic fibrosis (CF), yet these organisms compete under laboratory conditions. Recent observations that coinfection correlates with decreased health outcomes necessitate we develop a greater understanding of these interbacterial interactions. In this study, we tested the hypothesis that P. aeruginosa and/or S. aureus adopts phenotypes that allow coexistence during infection. We compared competitive interactions of P. aeruginosa and S. aureus isolates from mono- or coinfected CF patients employing in vitro coculture models. P. aeruginosa isolates from monoinfected patients were more competitive toward S. aureus than P. aeruginosa isolates from coinfected patients. We also observed that the least competitive P. aeruginosa isolates possessed a mucoid phenotype. Mucoidy occurs upon constitutive activation of the sigma factor AlgT/U, which regulates synthesis of the polysaccharide alginate and dozens of other secreted factors, including some previously described to kill S. aureus Here, we show that production of alginate in mucoid strains is sufficient to inhibit anti-S. aureus activity independent of activation of the AlgT regulon. Alginate reduces production of siderophores, 2-heptyl-4-hydroxyquinolone-N-oxide (HQNO), and rhamnolipids-each required for efficient killing of S. aureus These studies demonstrate alginate overproduction may be an important factor driving P. aeruginosa coinfection with S. aureusIMPORTANCE Numerous deep-sequencing studies have revealed the microbial communities present during respiratory infections in cystic fibrosis (CF) patients are diverse, complex, and dynamic. We now face the challenge of determining

  2. Oxidative stress-mediated antibacterial activity of graphene oxide and reduced graphene oxide in Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Gurunathan S

    2012-11-01

    Full Text Available Sangiliyandi Gurunathan, Jae Woong Han, Ahmed Abdal Dayem, Vasuki Eppakayala, Jin-Hoi KimDepartment of Animal Biotechnology, Konkuk University, Seoul, South KoreaBackground: Graphene holds great promise for potential use in next-generation electronic and photonic devices due to its unique high carrier mobility, good optical transparency, large surface area, and biocompatibility. The aim of this study was to investigate the antibacterial effects of graphene oxide (GO and reduced graphene oxide (rGO in Pseudomonas aeruginosa. In this work, we used a novel reducing agent, betamercaptoethanol (BME, for synthesis of graphene to avoid the use of toxic materials. To uncover the impacts of GO and rGO on human health, the antibacterial activity of two types of graphene-based material toward a bacterial model P. aeruginosa was studied and compared.Methods: The synthesized GO and rGO was characterized by ultraviolet-visible absorption spectroscopy, particle-size analyzer, X-ray diffraction, scanning electron microscopy and Raman spectroscopy. Further, to explain the antimicrobial activity of graphene oxide and reduced graphene oxide, we employed various assays, such as cell growth, cell viability, reactive oxygen species generation, and DNA fragmentation.Results: Ultraviolet-visible spectra of the samples confirmed the transition of GO into graphene. Dynamic light-scattering analyses showed the average size among the two types of graphene materials. X-ray diffraction data validated the structure of graphene sheets, and high-resolution scanning electron microscopy was employed to investigate the morphologies of prepared graphene. Raman spectroscopy data indicated the removal of oxygen-containing functional groups from the surface of GO and the formation of graphene. The exposure of cells to GO and rGO induced the production of superoxide radical anion and loss of cell viability. Results suggest that the antibacterial activities are contributed to by loss of

  3. Mitochondrial damage contributes to Pseudomonas aeruginosa activation of the inflammasome and is downregulated by autophagy.

    Science.gov (United States)

    Jabir, Majid Sakhi; Hopkins, Lee; Ritchie, Neil D; Ullah, Ihsan; Bayes, Hannah K; Li, Dong; Tourlomousis, Panagiotis; Lupton, Alison; Puleston, Daniel; Simon, Anna Katharina; Bryant, Clare; Evans, Thomas J

    2015-01-01

    The nucleotide-binding domain, leucine-rich repeat containing family caspase recruitment domain containing 4 (NLRC4) inflammasome can be activated by pathogenic bacteria via products translocated through the microbial type III secretion apparatus (T3SS). Recent work has shown that activation of the NLRP3 inflammasome is downregulated by autophagy, but the influence of autophagy on NLRC4 activation is unclear. We set out to determine how autophagy might influence this process, using the bacterium Pseudomonas aeruginosa, which activates the NLRC4 inflammasome via its T3SS. Infection resulted in T3SS-dependent mitochondrial damage with increased production of reactive oxygen intermediates and release of mitochondrial DNA. Inhibiting mitochondrial reactive oxygen release or degrading intracellular mitochondrial DNA abrogated NLRC4 inflammasome activation. Moreover, macrophages lacking mitochondria failed to activate NLRC4 following infection. Removal of damaged mitochondria by autophagy significantly attenuated NLRC4 inflammasome activation. Mitochondrial DNA bound specifically to NLRC4 immunoprecipitates and transfection of mitochondrial DNA directly activated the NLRC4 inflammasome; oxidation of the DNA enhanced this effect. Manipulation of autophagy altered the degree of inflammasome activation and inflammation in an in vivo model of P. aeruginosa infection. Our results reveal a novel mechanism contributing to NLRC4 activation by P. aeruginosa via mitochondrial damage and release of mitochondrial DNA triggered by the bacterial T3SS that is downregulated by autophagy.

  4. The potential of free cells of Pseudomonas aeruginosa on textile dye degradation.

    Science.gov (United States)

    Selvakumar, Kuppusamy Vaithilingam; Basha, Chiya Ahmed; Prabhu, Harinarayan Janardhana; Kalaichelvi, Ponnusamy; Nelliyan, Sudha

    2010-04-01

    The objective of the present work was to reduce chemical oxygen demand (COD), color of textile effluent containing dye Procion Blue 2G and recycle the treated effluent. To achieve this objective, the degradation potential of bacterial strain Pseudomonas aeruginosa was tested. In degradation, the 'clean' electro-oxidation (EO) process was combined with bio-treatment such that an electro-oxidation step was included between two bio-degradation treatments. Bio-oxidation process was carried out under aerobic and anoxic conditions. Results showed more than 90% reduction in COD and complete removal of color at the end of two cycles of combined oxidation process with a post electro-oxidation. The treated effluent was then subjected to photo-oxidation to remove the microbes so that the water can be recycled after the removal of total dissolved solids (TDS).

  5. PfeR, an enterobactin-responsive activator of ferric enterobactin receptor gene expression in Pseudomonas aeruginosa.

    OpenAIRE

    Dean, C. R.; Neshat, S; Poole, K

    1996-01-01

    PfeR (Regulator) and PfeS (Sensor), members of the superfamily of so-called two-component regulatory protein pairs, are required for the enterobactin-inducible production of the ferric enterobactin receptor (PfeA) in Pseudomonas aeruginosa. A pfeR knockout mutant failed to demonstrate enterobactin-inducible expression of a pfeA-lacZ fusion, indicating that PfeR acts at the level of pfeA gene expression. Consistent with this, PfeR overexpressed in P. aeruginosa bound, in bandshift assays, the ...

  6. Iminoguanidines as Allosteric Inhibitors of the Iron-Regulated Heme Oxygenase (HemO) of Pseudomonas aeruginosa

    OpenAIRE

    2016-01-01

    New therapeutic targets are required to combat multidrug resistant infections, such as the iron-regulated heme oxygenase (HemO) of Pseudomonas aeruginosa, due to links between iron and virulence and dependence on heme as an iron source during infection. Herein we report the synthesis and activity of a series of iminoguanidine-based inhibitors of HemO. Compound 23 showed a binding affinity of 5.7 µM and an MIC50 of 52.3 µg/mL against P. aeruginosa PAO1. An in cellulo activity assay was develop...

  7. Interspecies signalling via the Stenotrophomonas maltophilia diffusible signal factor influences biofilm formation and polymyxin tolerance in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Ryan, R.P.; Fouhy, Y.; Garcia, B.F.

    2008-01-01

    including the rhizosphere of plants and the cystic fibrosis lung. In mixed species biofilms, S. maltophilia substantially influenced the architecture of P. aeruginosa structures, which developed as extended filaments. This effect depended upon the synthesis of the diffusible signal factor (DSF) by S....... maltophilia and could be mimicked by the addition of synthetic DSF. This response of P. aeruginosa to DSF required PA1396, a sensor kinase with an input domain of related amino acid sequence to the sensory input domain of RpfC, which is responsible for DSF perception in xanthomonads. Mutation of PA1396...

  8. Physiological levels of nitrate support anoxic growth by denitrification of Pseudomonas aeruginosa at growth rates reported in cystic fibrosis lungs and sputum

    DEFF Research Database (Denmark)

    Klitten, Laura Line; Alhede, Morten; Kolpen, Mette;

    2014-01-01

    Chronic Pseudomonas aeruginosa lung infection is the most severe complication in patients with cystic fibrosis (CF). The infection is characterized by the formation of biofilm surrounded by numerous polymorphonuclear leukocytes (PMNs) and strong O2 depletion in the endobronchial mucus. We have...... reported that O2 is mainly consumed by the activated PMNs, while O2 consumption by aerobic respiration is diminutive and nitrous oxide (N2O) is produced in infected CF sputum. This suggests that the reported growth rates of P. aeruginosa in lungs and sputum may result from anaerobic respiration using...... comparable to our reported growth rates in the majority of P. aeruginosa cells in CF lungs and sputum. Thus, we have demonstrated that denitrification is required for P. aeruginosa growth in infected endobronchial CF mucus....

  9. Application of WGS data for O-specific antigen analysis and in silico serotyping of Pseudomonas aeruginosa isolates

    DEFF Research Database (Denmark)

    Thrane, Sandra Wingaard; Taylor, Véronique L.; Lund, Ole;

    2016-01-01

    Accurate typing methods are required for efficient infection control. The emergence of whole genome sequencing (WGS) technologies has enabled the development of genomics-based methods applicable for routine typing and surveillance of bacterial pathogens. In this study, we developed the Pseudomonas...... aeruginosa serotyper (PAst) program, which enabled in silico serotyping of P. aeruginosa isolates using WGS data. PAst has been made publically available as a web-service, and aptly facilitate high-throughput serotyping analysis. The program overcomes critical issues such as the loss of in vitro typeability....... This frequency is rarely achievable by conventional serotyping methods. The limited number of non-typeable isolates found using PAst was the result of either complete absence of OSA genes in the genomes or the artifact of genomic misassembly. With PAst, P. aeruginosa serotype data can be obtained from WGS...

  10. Proteomics of the oxidative stress response induced by hydrogen peroxide and paraquat reveals a novel AhpC-like protein in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Hare, Nathan J; Scott, Nichollas E; Shin, Eun Hye H;

    2011-01-01

    . aeruginosa establishes infection in the cystic fibrosis lung and eventually out-competes other pathogenic bacteria to persist into chronic infection. We utilized a proteomics approach to identify the proteins associated with the oxidative stress response of P. aeruginosa PAO1 to hydrogen peroxide...... by hypothetical protein PA3529 following treatment with 10 mM H(2) O(2) . AhpC belongs to the 2-Cys peroxiredoxin family and is a redox enzyme responsible for removing peroxides in bacterial cells. MS analysis showed that PA3529 was altered by the formation of a dimer via a disulfide bond in a manner analogous...

  11. Chronic Pseudomonas aeruginosa lung infection in normal and athymic rats

    DEFF Research Database (Denmark)

    Johansen, H K; Espersen, F; Pedersen, S S

    1993-01-01

    We have compared a chronic lung infection with Pseudomonas aeruginosa embedded in alginate beads in normal and athymic rats with an acute infection with free live P. aeruginosa bacteria. The following parameters were observed and described: mortality, macroscopic and microscopic pathologic change...

  12. Effects of ginseng on Pseudomonas aeruginosa motility and biofilm formation

    DEFF Research Database (Denmark)

    Wu, Hong; Lee, Baoleri; Yang, Liang

    2011-01-01

    Biofilm-associated chronic Pseudomonas aeruginosa lung infections in patients with cystic fibrosis are virtually impossible to eradicate with antibiotics because biofilm-growing bacteria are highly tolerant to antibiotics and host defense mechanisms. Previously, we found that ginseng treatments......-associated chronic infections caused by P. aeruginosa....

  13. Antibacterial activity of five Peruvian medicinal plants against Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    Gabriela; Ulloa-Urizar; Miguel; Angel; Aguilar-Luis; María; del; Carmen; De; Lama-Odría; José; Camarena-Lizarzaburu; Juana; del; Valle; Mendoza

    2015-01-01

    Objective: To evaluate the susceptibility of Pseudomonas aeruginosa(P. aeruginosa)in vitro to the ethanolic extracts obtained from five different Peruvian medicinal plants.Methods: The plants were chopped and soaked in absolute ethanol(1:2, w/v). The antibacterial activity of compounds against P. aeruginosa was evaluated using the cupplate agar diffusion method.Results: The extracts from Maytenus macrocarpa("Chuchuhuasi"), Dracontium loretense Krause("Jergon Sacha"), Tabebuia impetiginosa("Tahuari"), Eucalyptus camaldulensis Dehn(eucalyptus), Uncaria tomentosa("U?a de gato") exhibited favorable antibacterial activity against P. aeruginosa. The inhibitory effect of the extracts on the strains of P. aeruginosa tested demonstrated that Tabebuia impetiginosa and Maytenus macrocarpa possess higher antibacterial activity.Conclusions: The results of the present study scientifically validate the inhibitory capacity of the five medicinal plants attributed by their common use in folk medicine and contribute towards the development of new treatment options based on natural products.

  14. Antibacterial activity of ifve Peruvian medicinal plants against Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    Gabriela Ulloa-Urizar; Miguel Angel Aguilar-Luis; Mara del Carmen De Lama-Odra; Jos Camarena-Lizarzaburu; Juana del Valle Mendoza

    2015-01-01

    Objective:To evaluate the susceptibility of Pseudomonas aeruginosa (P. aeruginosa) in vitro to the ethanolic extracts obtained from five different Peruvian medicinal plants. Methods:The plants were chopped and soaked in absolute ethanol (1:2, w/v). The antibacterial activity of compounds against P. aeruginosa was evaluated using the cup-plate agar diffusion method. Results:The extracts from Maytenus macrocarpa (“Chuchuhuasi”), Dracontium loretense Krause (“Jergon Sacha”), Tabebuia impetiginosa (“Tahuari”), Eucalyptus camaldulensis Dehn (eucalyptus), Uncaria tomentosa (“Uña de gato”) exhibited favorable antibacterial activity against P. aeruginosa. The inhibitory effect of the extracts on the strains of P. aeruginosa tested demonstrated that Tabebuia impetiginosa and Maytenus macrocarpa possess higher antibacterial activity. Conclusions:The results of the present study scientifically validate the inhibitory capacity of the five medicinal plants attributed by their common use in folk medicine and contribute towards the development of new treatment options based on natural products.

  15. Ga@C-dots as an antibacterial agent for the eradication of Pseudomonas aeruginosa

    Science.gov (United States)

    Kumar, Vijay Bhooshan; Natan, Michal; Jacobi, Gila; Porat, Ze’ev; Banin, Ehud; Gedanken, Aharon

    2017-01-01

    The opportunistic pathogen Pseudomonas aeruginosa causes infections that are difficult to treat by antibiotic therapy. This research article reports on the synthesis of gallium (Ga) doped in carbon (C)-dots (Ga@C-dots) and their antimicrobial activity against free-living P. aeruginosa bacteria. The synthesis of Ga@C-dots was carried out by sonicating molten Ga (for 2.5 h) in polyethylene glycol-400, which acts as both a medium and carbon source. The resultant Ga@C-dots, having an average diameter of 9±2 nm, showed remarkably enhanced antibacterial activity compared with undoped C-dots. This was reflected by the much lower concentration of Ga doped within Ga@C-dots which was required for full inhibition of the bacterial growth. These results highlight the possibility of using Ga@C-dots as potential antimicrobial agents. PMID:28176980

  16. Resistance of Pseudomonas aeruginosa to liquid disinfectants on contaminated surfaces before formation of biofilms.

    Science.gov (United States)

    Sagripanti, J L; Bonifacino, A

    2000-01-01

    A comparison was made of the effectiveness of popular disinfectants (Cavicide, Cidexplus, Clorox, Exspor, Lysol, Renalin, and Wavicide) under conditions prescribed for disinfection in the respective product labels on Pseudomonas aeruginosa either in suspension or deposited onto surfaces of metallic or polymeric plastic devices. The testing also included 7 nonformulated germicidal agents (glutaraldehyde, formaldehyde, peracetic acid, hydrogen peroxide, sodium hypochlorite, phenol, and cupric ascorbate) commonly used in disinfection and decontamination. Results showed that P. aeruginosa is on average 300-fold more resistant when present on contaminated surfaces than in suspension. This increase in resistance agrees with results reported in studies of biofilms, but unexpectedly, it precedes biofilm formation. The surface to which bacteria are attached can influence the effectiveness of disinfectants. Viable bacteria attached to devices may require dislodging through more than a one-step method for detection. The data, obtained with a sensitive and quantitative test, suggest that disinfectants are less effective on contaminated surfaces than generally acknowledged.

  17. Biotransformation of myrcene by Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Hashemi Elham

    2011-05-01

    Full Text Available Abstract Background Dihydrolinalool and terpineol are sources of fragrances that provide a unique volatile terpenoid alcohol of low toxicity and thus are widely used in the perfumery industry, in folk medicine, and in aromatherapy. They are important chemical constituents of the essential oil of many plants. Previous studies have concerned the biotransformation of limonene by Pseudomonas putida. The objective of this research was to study biotransformation of myrcene by Pseudomonas aeruginosa. The culture preparation was done using such variables as different microbial methods and incubation periods to obtain maximum cells of P. aeruginosa for myrcene biotransformation. Results It was found that myrcene was converted to dihydrolinalool and 2,6-dimethyloctane in high percentages. The biotransformation products were identified by Fourier-transform infrared spectroscopy (FT-IR, ultraviolet (UV analysis, gas chromatography (GC, and gas chromatography-mass spectroscopy (GC-MS. Comparison of the different incubation times showed that 3 days was more effective, the major products being 2,6-dimethyloctane (90.0% and α-terpineol (7.7% and comprising 97.7%. In contrast, the main compounds derived for an incubation time of 1.5 days were dihydrolinalool (79.5% and 2,6-dimethyloctane (9.3%, with a total yield of 88.8%.

  18. Pseudomonas aeruginosa ventilator-associated pneumonia management

    Directory of Open Access Journals (Sweden)

    Ramírez-Estrada S

    2016-01-01

    Full Text Available Sergio Ramírez-Estrada,1 Bárbara Borgatta,1,2 Jordi Rello3,4 1Critical Care Department, Vall d'Hebron University Hospital, 2CRIPS, Vall d'Hebron Institute of Research (VHIR, 3Department of Medicine, Universitat Autònoma de Barcelona (UAB, Barcelona, 4Centro de Investigación Biomédica en Red Enfermedad Respiratoria – CIBERES, Madrid, Spain Abstract: Ventilator-associated pneumonia is the most common infection in intensive care unit patients associated with high morbidity rates and elevated economic costs; Pseudomonas aeruginosa is one of the most frequent bacteria linked with this entity, with a high attributable mortality despite adequate treatment that is increased in the presence of multiresistant strains, a situation that is becoming more common in intensive care units. In this manuscript, we review the current management of ventilator-associated pneumonia due to P. aeruginosa, the most recent antipseudomonal agents, and new adjunctive therapies that are shifting the way we treat these infections. We support early initiation of broad-spectrum antipseudomonal antibiotics in present, followed by culture-guided monotherapy de-escalation when susceptibilities are available. Future management should be directed at blocking virulence; the role of alternative strategies such as new antibiotics, nebulized treatments, and vaccines is promising. Keywords: multidrug-resistant, ICU, new-antibiotics, adjunctive-therapies, care-bundles

  19. Cockayne syndrome: varied requirement of transcription-coupled nucleotide excision repair for the removal of three structurally different adducts from transcribed DNA.

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    Nataliya Kitsera

    Full Text Available Hereditary defects in the transcription-coupled nucleotide excision repair (TC-NER pathway of damaged DNA cause severe neurodegenerative disease Cockayne syndrome (CS, however the origin and chemical nature of the underlying DNA damage had remained unknown. To find out, to which degree the structural properties of DNA lesions determine the extent of transcription arrest in human CS cells, we performed quantitative host cell reactivation analyses of expression vectors containing various synthetic adducts. We found that a single 3-(deoxyguanosin-N2-yl-2-acetylaminofluorene adduct (dG(N2-AAF constitutes an unsurmountable obstacle to transcription in both CS-A and CS-B cells and is removed exclusively by the CSA- and CSB-dependent pathway. In contrast, contribution of the CS proteins to the removal of two other transcription-blocking DNA lesions - N-(deoxyguanosin-8-yl-2-acetylaminofluorene (dG(C8-AAF and cyclobutane thymine-thymine (TT dimer - is only minor (TT dimer or none (dG(C8-AAF. The unique properties of dG(N2-AAF identify this adduct as a prototype for a new class of DNA lesions that escape the alternative global genome repair and could be critical for the CS pathogenesis.

  20. Removal of cyanobacteria and cyanotoxins from drinking water by powdered activated carbon adsorption/ultrafiltration

    OpenAIRE

    Campinas, Margarida Páscoa

    2009-01-01

    Tese dout., Ciências e Tecnologias do Ambiente, Universidade do Algarve, 2009 PAC/UF was investigated to remove the cyanobacterium Microcysis aeruginosa and microcystins, focusing on toxins adsorption onto PAC and the combined effect of the water organic and inorganic matrices, the cells removal and lysis by UF, and PAC contribution to membrane fouling control and microcystins removal by PAC/UF. The fine-grade mesoporous PAC presented high capacity and fast kinetics for microc...

  1. Tattoo removal.

    Science.gov (United States)

    Adatto, Maurice A; Halachmi, Shlomit; Lapidoth, Moshe

    2011-01-01

    Over 50,000 new tattoos are placed each year in the United States. Studies estimate that 24% of American college students have tattoos and 10% of male American adults have a tattoo. The rising popularity of tattoos has spurred a corresponding increase in tattoo removal. Not all tattoos are placed intentionally or for aesthetic reasons though. Traumatic tattoos due to unintentional penetration of exogenous pigments can also occur, as well as the placement of medical tattoos to mark treatment boundaries, for example in radiation therapy. Protocols for tattoo removal have evolved over history. The first evidence of tattoo removal attempts was found in Egyptian mummies, dated to have lived 4,000 years BC. Ancient Greek writings describe tattoo removal with salt abrasion or with a paste containing cloves of white garlic mixed with Alexandrian cantharidin. With the advent of Q-switched lasers in the late 1960s, the outcomes of tattoo removal changed radically. In addition to their selective absorption by the pigment, the extremely short pulse duration of Q-switched lasers has made them the gold standard for tattoo removal.

  2. Contributions of efflux pumps to high level resistance of Pseudomonas aeruginosa to ciprofloxacin

    Institute of Scientific and Technical Information of China (English)

    WANG Dan-dan; SUN Tie-ying; HU Yun-jian

    2007-01-01

    @@ Pseudomonas aeruginosa (P. aeruginosa) is one of the leading pathogens involved in nosocomial pneumonia. In addition, P. aeruginosa infection is associated with significant morbidity and mortality.1 A major problem in P. aeruginosa infection is that this organism exhibits natural and acquired resistance to many structurally and functionally diverse antibiotics.

  3. Tobramycin at subinhibitory concentration inhibits the RhlI/R quorum sensing system in a Pseudomonas aeruginosa environmental isolate

    Directory of Open Access Journals (Sweden)

    Venturi Vittorio

    2010-06-01

    Full Text Available Abstract Background Antibiotics are not only small molecules with therapeutic activity in killing or inhibiting microbial growth, but can also act as signaling molecules affecting gene expression in bacterial communities. A few studies have demonstrated the effect of tobramycin as a signal molecule on gene expression at the transcriptional level and its effect on bacterial physiology and virulence. These have shown that subinhibitory concentrations (SICs of tobramycin induce biofilm formation and enhance the capabilities of P. aeruginosa to colonize specific environments. Methods Environmental P. aeruginosa strain PUPa3 was grown in the presence of different concentrations of tobramycin and it was determined at which highest concentration SIC, growth, total protein levels and translation efficiency were not affected. At SIC it was then established if phenotypes related to cell-cell signaling known as quorum sensing were altered. Results In this study it was determined whether tobramycin sensing/response at SICs was affecting the two independent AHL QS systems in an environmental P. aeruginosa strain. It is reasonable to assume that P. aeruginosa encounters tobramycin in nature since it is produced by niche mate Streptomyces tenebrarius. It was established that SICs of tobramycin inhibited the RhlI/R system by reducing levels of C4-HSL production. This effect was not due to a decrease of rhlI transcription and required tobramycin-ribosome interaction. Conclusions Tobramycin signaling in P. aeruginosa occurs and different strains can have a different response. Understanding the tobramycin response by an environmental P. aeruginosa will highlight possible inter-species signalling taking place in nature and can possible also have important implications in the mode of utilization for human use of this very important antibiotic.

  4. Enantioselective changes in oxidative stress and toxin release in Microcystis aeruginosa exposed to chiral herbicide diclofop acid

    Energy Technology Data Exchange (ETDEWEB)

    Ye, Jing [School of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418 (China); MOE Key Lab of Environmental Remediation and Ecosystem Health, College of Natural Research and Environmental Sciences, Zhejiang University, Hangzhou 310058 (China); Zhang, Ying [Department of Environmental Science, East China Normal University, Shanghai 200241 (China); Chen, Shengwen [School of Urban Development and Environment Engineering, Shanghai Second Polytechnic University, Shanghai 201209 (China); Liu, Chaonan; Zhu, Yongqiang [School of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418 (China); Liu, Weiping, E-mail: wliu@zju.edu.cn [MOE Key Lab of Environmental Remediation and Ecosystem Health, College of Natural Research and Environmental Sciences, Zhejiang University, Hangzhou 310058 (China)

    2014-01-15

    Highlights: •The first study on enantioselective oxidative stress and toxin release from Microcystis aeruginosa. •Provide information for the R-enantiomer poses more oxidative stress than the S-enantiomer. •Lifecycle analysis of chiral pollutants needs more attention in environmental assessment. -- Abstract: Enantioselective oxidative stress and toxin release from Microcystis aeruginosa after exposure to the chiral herbicide diclofop acid were investigated. Racemic diclofop acid, R-diclofop acid and S-diclofop acid induced reactive oxygen species (ROS) generation, increased the concentration of malondialdehyde (MDA), enhanced the activity of superoxide dismutase (SOD) and triggered toxin release in M. aeruginosa to varying degrees. The increase in MDA concentration and SOD activity in M. aeruginosa occurred sooner after exposure to diclofop acid than when the cyanobacteria was exposed to either the R- and the S-enantiomer. In addition, enantioselective toxicity of the enantiomers was observed. The R-enantiomer trigged more ROS generation, more SOD activity and more toxin synthesis and release in M. aeruginosa cells than the S-enantiomer. Diclofop acid and its R-enantiomer may collapse the transmembrane proton gradient and destroy the cell membrane through lipid peroxidation and free radical oxidation, whereas the S-enantiomer did not demonstrate such action. R-diclofop acid inhibits the growth of M. aeruginosa in the early stage, but ultimately induced greater toxin release, which has a deleterious effect on the water column. These results indicate that more comprehensive study is needed to determine the environmental safety of the enantiomers, and application of chiral pesticides requires more direct supervision and training. Additionally, lifecycle analysis of chiral pollutants in aquatic system needs more attention to aide in the environmental assessment of chiral pesticides.

  5. Berberine is a novel type efflux inhibitor which attenuates the MexXY-mediated aminoglycoside resistance in Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Yuji Morita

    2016-08-01

    Full Text Available The emergence and spread of multidrug-resistant P. aeruginosa infections is of great concern, as very few agents are effective against strains of this species. Methanolic extracts from the Coptidis Rhizoma (the rhizomes of Coptis japonica var. major Satake or Phellodendri Cortex (the bark of Phellodendron chinense Schneider markedly reduced resistance to anti-pseudomonal aminoglycosides (e.g. amikacin in multidrug-resistant P. aeruginosa strains. Berberine, the most abundant benzylisoquinoline alkaloid in the two extracts, reduced aminoglycoside resistance of P. aeruginosa via a mechanism that required the MexXY multidrug efflux system; berberine also reduced aminoglycoside MICs in Achromobacter xylosoxidans and Burkholderia cepacia, two species that harbor intrinsic multidrug efflux systems very similar to the MexXY. Furthermore this compound inhibited MexXY-dependent antibiotic resistance of other classes including cephalosporins (cefepime, macrolides (erythromycin, and lincosamides (lincomycin demonstrated using a pseudomonad lacking the 4 other major Mex pumps. Although phenylalanine-arginine beta-naphthylamide (PAβN, a well-known efflux inhibitor, antagonized aminoglycoside in a MexXY-dependent manner, a lower concentration of berberine was sufficient to reduce amikacin resistance of P. aeruginosa in the presence of PAβN. Moreover, berberine enhanced the synergistic effects of amikacin and piperacillin (and vice versa in multidrug-resistant P. aeruginosa strains. Thus, berberine appears to be a novel type inhibitor of the MexXY-dependent aminoglycoside efflux in P. aeruginosa. As aminoglycosides are molecules of choice to treat severe infections the clinical impact is potentially important.

  6. Berberine Is a Novel Type Efflux Inhibitor Which Attenuates the MexXY-Mediated Aminoglycoside Resistance in Pseudomonas aeruginosa

    Science.gov (United States)

    Morita, Yuji; Nakashima, Ken-ichi; Nishino, Kunihiko; Kotani, Kenta; Tomida, Junko; Inoue, Makoto; Kawamura, Yoshiaki

    2016-01-01

    The emergence and spread of multidrug-resistant P. aeruginosa infections is of great concern, as very few agents are effective against strains of this species. Methanolic extracts from the Coptidis Rhizoma (the rhizomes of Coptis japonica var. major Satake) or Phellodendri Cortex (the bark of Phellodendron chinense Schneider) markedly reduced resistance to anti-pseudomonal aminoglycosides (e.g., amikacin) in multidrug-resistant P. aeruginosa strains. Berberine, the most abundant benzylisoquinoline alkaloid in the two extracts, reduced aminoglycoside resistance of P. aeruginosa via a mechanism that required the MexXY multidrug efflux system; berberine also reduced aminoglycoside MICs in Achromobacter xylosoxidans and Burkholderia cepacia, two species that harbor intrinsic multidrug efflux systems very similar to the MexXY. Furthermore this compound inhibited MexXY-dependent antibiotic resistance of other classes including cephalosporins (cefepime), macrolides (erythromycin), and lincosamides (lincomycin) demonstrated using a pseudomonad lacking the four other major Mex pumps. Although phenylalanine-arginine beta-naphthylamide (PAβN), a well-known efflux inhibitor, antagonized aminoglycoside in a MexXY-dependent manner, a lower concentration of berberine was sufficient to reduce amikacin resistance of P. aeruginosa in the presence of PAβN. Moreover, berberine enhanced the synergistic effects of amikacin and piperacillin (and vice versa) in multidrug-resistant P. aeruginosa strains. Thus, berberine appears to be a novel type inhibitor of the MexXY-dependent aminoglycoside efflux in P. aeruginosa. As aminoglycosides are molecules of choice to treat severe infections the clinical impact is potentially important. PMID:27547203

  7. Candida albicans ethanol stimulates Pseudomonas aeruginosa WspR-controlled biofilm formation as part of a cyclic relationship involving phenazines.

    Directory of Open Access Journals (Sweden)

    Annie I Chen

    2014-10-01

    Full Text Available In chronic infections, pathogens are often in the presence of other microbial species. For example, Pseudomonas aeruginosa is a common and detrimental lung pathogen in individuals with cystic fibrosis (CF and co-infections with Candida albicans are common. Here, we show that P. aeruginosa biofilm formation and phenazine production were strongly influenced by ethanol produced by the fungus C. albicans. Ethanol stimulated phenotypes that are indicative of increased levels of cyclic-di-GMP (c-di-GMP, and levels of c-di-GMP were 2-fold higher in the presence of ethanol. Through a genetic screen, we found that the diguanylate cyclase WspR was required for ethanol stimulation of c-di-GMP. Multiple lines of evidence indicate that ethanol stimulates WspR signaling through its cognate sensor WspA, and promotes WspR-dependent activation of Pel exopolysaccharide production, which contributes to biofilm maturation. We also found that ethanol stimulation of WspR promoted P. aeruginosa colonization of CF airway epithelial cells. P. aeruginosa production of phenazines occurs both in the CF lung and in culture, and phenazines enhance ethanol production by C. albicans. Using a C. albicans adh1/adh1 mutant with decreased ethanol production, we found that fungal ethanol strongly altered the spectrum of P. aeruginosa phenazines in favor of those that are most effective against fungi. Thus, a feedback cycle comprised of ethanol and phenazines drives this polymicrobial interaction, and these relationships may provide insight into why co-infection with both P. aeruginosa and C. albicans has been associated with worse outcomes in cystic fibrosis.

  8. Rapid conversion of Pseudomonas aeruginosa to a spherical cell morphotype facilitates tolerance to carbapenems and penicillins but increases susceptibility to antimicrobial peptides.

    Science.gov (United States)

    Monahan, Leigh G; Turnbull, Lynne; Osvath, Sarah R; Birch, Debra; Charles, Ian G; Whitchurch, Cynthia B

    2014-01-01

    The Gram-negative human pathogen Pseudomonas aeruginosa tolerates high concentrations of β-lactam antibiotics. Despite inhibiting the growth of the organism, these cell wall-targeting drugs exhibit remarkably little bactericidal activity. However, the mechanisms underlying β-lactam tolerance are currently unclear. Here, we show that P. aeruginosa undergoes a rapid en masse transition from normal rod-shaped cells to viable cell wall-defective spherical cells when treated with β-lactams from the widely used carbapenem and penicillin classes. When the antibiotic is removed, the entire population of spherical cells quickly converts back to the normal bacillary form. Our results demonstrate that these rapid population-wide cell morphotype transitions function as a strategy to survive antibiotic exposure. Taking advantage of these findings, we have developed a novel approach to efficiently kill P. aeruginosa by using carbapenem treatment to induce en masse transition to the spherical cell morphotype and then exploiting the relative fragility and sensitivity of these cells to killing by antimicrobial peptides (AMPs) that are relatively inactive against P. aeruginosa bacillary cells. This approach could broaden the repertoire of antimicrobial compounds used to treat P. aeruginosa and serve as a basis for developing new therapeutic agents to combat bacterial infections.

  9. A Phytoanticipin Derivative, Sodium Houttuyfonate, Induces in Vitro Synergistic Effects with Levofloxacin against Biofilm Formation by Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Jing Shao

    2012-09-01

    Full Text Available Antibiotic resistance has become the main deadly factor in infections, as bacteria can protect themselves by hiding in a self-constructed biofilm. Consequently, more attention is being paid to the search for “non-antibiotic drugs” to solve this problem. Phytoanticipins, the natural antibiotics from plants, could be a suitable alternative, but few works on this aspect have been reported. In this study, a preliminary study on the synergy between sodium houttuyfonate (SH and levofloxacin (LFX against the biofilm formation of Pseudomonas aeruginosa was performed. The minimal inhibitory concentrations (MIC of LFX and SH, anti-biofilm formation and synergistic effect on Pseudomonas aeruginosa, and quantification of alginate were determined by the microdilution method, crystal violet (CV assay, checkerboard method, and hydroxybiphenyl colorimetry. The biofilm morphology of Pseudomonas aeruginosa was observed by fluorescence microscope and scanning electric microscope (SEM. The results showed that: (i LFX and SH had an obvious synergistic effect against Pseudomonas aeruginosa with MIC values of 0.25 μg/mL and 128 μg/mL, respectively; (ii ½ × MIC SH combined with 2 × MIC LFX could suppress the biofilm formation of Pseudomonas aeruginosa effectively, with up to 73% inhibition; (iii the concentration of alginate decreased dramatically by a maximum of 92% after treatment with the combination of antibiotics; and (iv more dead cells by fluorescence microscope and more removal of extracellular polymeric structure (EPS by SEM were observed after the combined treatment of LFX and SH. Our experiments demonstrate the promising future of this potent antimicrobial agent against biofilm-associated infections.

  10. Sequencing and Characterization of Pseudomonas aeruginosa phage JG004

    Directory of Open Access Journals (Sweden)

    Bunk Boyke

    2011-05-01

    Full Text Available Abstract Background Phages could be an important alternative to antibiotics, especially for treatment of multiresistant bacteria as e.g. Pseudomonas aeruginosa. For an effective use of bacteriophages as antimicrobial agents, it is important to understand phage biology but also genes of the bacterial host essential for phage infection. Results We isolated and characterized a lytic Pseudomonas aeruginosa phage, named JG004, and sequenced its genome. Phage JG004 is a lipopolysaccharide specific broad-host-range phage of the Myoviridae phage family. The genome of phage JG004 encodes twelve tRNAs and is highly related to the PAK-P1 phage genome. To investigate phage biology and phage-host interactions, we used transposon mutagenesis of the P. aeruginosa host and identified P. aeruginosa genes, which are essential for phage infection. Analysis of the respective P. aeruginosa mutants revealed several characteristics, such as host receptor and possible spermidine-dependance of phage JG004. Conclusions Whole genome sequencing of phage JG004 in combination with identification of P. aeruginosa host genes essential for infection, allowed insights into JG004 biology, revealed possible resistance mechanisms of the host bacterium such as mutations in LPS and spermidine biosynthesis and can also be used to characterize unknown gene products in P. aeruginosa.

  11. Characteristic changes in algal organic matter derived from Microcystis aeruginosa in microbial fuel cells.

    Science.gov (United States)

    Wang, Huan; Lu, Lu; Liu, Dongmei; Cui, Fuyi; Wang, Peng

    2015-11-01

    The objective of this study was to investigate behavior of algal organic matter (AOM) during bioelectrochemical oxidation in microbial fuel cell in terms of compositions and structures. Study revealed that the AOM derived from blue-green algae Microcystis aeruginosa could be degraded more completely (82% COD removal) in microbial fuel cells (MFCs) than by anaerobic fermentation (24% COD removal) in a control reactor without closed-circuit electrode and electricity was produced simultaneously. A variety of techniques were used to characterize the changes in AOM compositions and structures during bioelectrochemical oxidation. The presence of syntrophic interactions between electrochemical active bacteria and fermentative bacteria to degrade large molecular organics into small molecular substances, which could be oxidized by electrode but not by fermentation. The dominant tryptophan protein-like substances, humic acid-like substances and Chlorophyll a in AOM were highly degraded during MFC treatment.

  12. Comparison of UVB and UVC irradiation disinfection efficacies on Pseudomonas Aeruginosa (P. aeruginosa) biofilm

    Science.gov (United States)

    Argyraki, A.; Markvart, M.; Nielsen, Anne; Bjarnsholt, T.; Bjørndal, L.; Petersen, P. M.

    2016-04-01

    Disinfection routines are important in all clinical applications. The uprising problem of antibiotic resistance has driven major research efforts towards alternative disinfection approaches, involving light-based solutions. Pseudomonas aeruginosa (P. aeruginosa) is a common bacterium that can cause skin, soft tissue, lungs, kidney and urinary tract infections. Moreover, it can be found on and in medical equipment causing often cross infections in hospitals. The objective of this study was to test the efficiency, of two different light-based disinfection treatments, namely UVB and UVC irradiation, on P. aeruginosa biofilms at different growth stages. In our experiments a new type of UV light emitting diodes (LEDs) were used to deliver UV irradiation on the biofilms, in the UVB (296nm) and UVC (266nm) region. The killing rate was studied as a function of dose for 24h grown biofilms. The dose was ramped from 72J/m2 to 10000J/m2. It was shown that UVB irradiation was more effective than UVC irradiation in inactivating P. aeruginosa biofilms. No colony forming units (CFU) were observed for the UVB treated biofilms when the dose was 10000 J/m2 (CFU in control sample: 7.5 x 104). UVB irradiation at a dose of 20000J/m2 on mature biofilms (72h grown) resulted in a 3.9 log killing efficacy. The fact that the wavelength of 296nm exists in daylight and has such disinfection ability on biofilms gives new perspectives for applications within disinfection at hospitals.

  13. Genetic Screen Reveals Link between the Maternal Effect Sterile Gene mes-1 and Pseudomonas aeruginosa-induced Neurodegeneration in Caenorhabditis elegans.

    Science.gov (United States)

    Wu, Qiuli; Cao, Xiou; Yan, Dong; Wang, Dayong; Aballay, Alejandro

    2015-12-01

    Increasing evidence indicates that immune responses to microbial infections may contribute to neurodegenerative diseases. Here, we show that Pseudomonas aeruginosa infection of Caenorhabditis elegans causes a number of neural changes that are hallmarks of neurodegeneration. Using an unbiased genetic screen to identify genes involved in the control of P. aeruginosa-induced neurodegeneration, we identified mes-1, which encodes a receptor tyrosine kinase-like protein that is required for unequal cell divisions in the early embryonic germ line. We showed that sterile but not fertile mes-1 animals were resistant to neurodegeneration induced by P. aeruginosa infection. Similar results were observed using animals carrying a mutation in the maternal effect gene pgl-1, which is required for postembryonic germ line development, and the germ line-deficient strains glp-1 and glp-4. Additional studies indicated that the FOXO transcription factor DAF-16 is required for resistance to P. aeruginosa-induced neurodegeneration in germ line-deficient strains. Thus, our results demonstrate that P. aeruginosa infection results in neurodegeneration phenotypes in C. elegans that are controlled by the germ line in a cell-nonautonomous manner.

  14. Hair removal

    DEFF Research Database (Denmark)

    Haedersdal, Merete; Haak, Christina S

    2011-01-01

    Hair removal with optical devices has become a popular mainstream treatment that today is considered the most efficient method for the reduction of unwanted hair. Photothermal destruction of hair follicles constitutes the fundamental concept of hair removal with red and near-infrared wavelengths...... suitable for targeting follicular and hair shaft melanin: normal mode ruby laser (694 nm), normal mode alexandrite laser (755 nm), pulsed diode lasers (800, 810 nm), long-pulse Nd:YAG laser (1,064 nm), and intense pulsed light (IPL) sources (590-1,200 nm). The ideal patient has thick dark terminal hair......, white skin, and a normal hormonal status. Currently, no method of lifelong permanent hair eradication is available, and it is important that patients have realistic expectations. Substantial evidence has been found for short-term hair removal efficacy of up to 6 months after treatment with the available...

  15. Characterization of a Heme-Regulated Non-Coding RNA Encoded by the prrF Locus of Pseudomonas aeruginosa

    OpenAIRE

    Oglesby-Sherrouse, Amanda G.; Vasil, Michael L.

    2010-01-01

    Pseudomonas aeruginosa, an opportunistic pathogen, requires iron for virulence and can obtain this nutrient via the acquisition of heme, an abundant source of iron in the human body. A surplus of either iron or heme can lead to oxidative stress; thus, the Fur (ferric uptake regulator) protein blocks expression of genes required for iron and heme uptake in iron-replete environments. Fur also represses expression of two nearly identical genes encoding the 116- and 114-nucleotide (nt) long PrrF1...

  16. Positively charged biopolymeric nanoparticles for the inhibition of Pseudomonas aeruginosa biofilms

    Science.gov (United States)

    Chronopoulou, Laura; Di Domenico, Enea Gino; Ascenzioni, Fiorentina; Palocci, Cleofe

    2016-10-01

    Currently, many microbial infections have the potential to become lethal owing to the development of antimicrobial resistance by means of different mechanisms and mainly on the basis of the fact that many drugs are unable to reach therapeutic levels in the target sites. This requires the use of high doses and frequent administrations, causing adverse side effects or in some cases toxicity. The use of nanoparticle systems could help overcome such problems and increase drug efficacy. In the present study, we developed a new drug delivery system based on the use of biopolymeric nanovectors loaded with tobramycin (Tb), which is the standard antibiotic for the treatment of Cystic Fibrosis-associated P. aeruginosa lung infections. Tb-loaded biopolymeric nanoparticles composed by dextran sulfate (DS) and chitosan (CS) were prepared by ionotropic gelation. We optimized drug entrapment in DS/CS nanoparticles, obtaining particles of 170 nm and with a drug loading of 400 µg Tb/mg of nanoparticles. In accord with in vitro release experiments, such preparations were able to release approximately 25 % of their cargo in 60 h. In vitro, the antimicrobial efficacy of the drug delivery system on P. aeruginosa biofilm was tested and compared to the effects of free drug revealing that this formulation can reduce the viability of P. aeruginosa biofilms for 48 h with a single-dose administration.

  17. Ga@C-dots as an antibacterial agent for the eradication of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Kumar VB

    2017-01-01

    Full Text Available Vijay Bhooshan Kumar,1 Michal Natan,2 Gila Jacobi,2 Ze’ev Porat,3,4 Ehud Banin,2 Aharon Gedanken1,5 1Department of Chemistry, 2Mina and Everard Goodman Faculty of Life Sciences, Bar Ilan Institute for Nanotechnology and Advanced Materials Bar-Ilan University, Ramat Gan, 3Division of Chemistry, Nuclear Research Center-Negev, 4Institutes of Applied Research, Ben-Gurion University of the Negev, Be’er-Sheva, Israel; 5National Cheng Kung University, Department of Materials Science and Engineering, Tainan, Taiwan Abstract: The opportunistic pathogen Pseudomonas aeruginosa causes infections that are difficult to treat by antibiotic therapy. This research article reports on the synthesis of gallium (Ga doped in carbon (C-dots (Ga@C-dots and their antimicrobial activity against free-living P. aeruginosa bacteria. The synthesis of Ga@C-dots was carried out by sonicating molten Ga (for 2.5 h in polyethylene glycol-400, which acts as both a medium and carbon source. The resultant Ga@C-dots, having an average diameter of 9±2 nm, showed remarkably enhanced antibacterial activity compared with undoped C-dots. This was reflected by the much lower concentration of Ga doped within Ga@C-dots which was required for full inhibition of the bacterial growth. These results highlight the possibility of using Ga@C-dots as potential antimicrobial agents. Keywords: C-dots, Ga@C-dots, sonochemistry, gallium, antibacterial, Pseudomonas aeruginosa

  18. Siderophore as a potential plant growth-promoting agent produced by Pseudomonas aeruginosa JAS-25.

    Science.gov (United States)

    Sulochana, M B; Jayachandra, S Y; Kumar, S Anil; Parameshwar, A B; Reddy, K Mohan; Dayanand, A

    2014-09-01

    Siderophores scavenges Fe(+3) from the vicinity of the roots of plants, and thus limit the amount of iron required for the growth of pathogens such as Fusarium oxysporum, Pythium ultimum, and Fusarium udum, which cause wilt and root rot disease in crops. The ability of Pseudomonas to grow and to produce siderophore depends upon the iron content, pH, and temperature. Maximum yield of siderophore of 130 μM was observed at pH 7.0 ± 0.2 and temperature of 30 °C at 30 h. The threshold level of iron was 50 μM, which increases up to 150 μM, favoring growth but drastically affecting the production of siderophore by Pseudomonas aeruginosa JAS-25. The seeds of agricultural crops like Cicer arietinum (chick pea), Cajanus cajan (pigeon pea), and Arachis hypogaea (ground nut) were treated with P. aeruginosa JAS-25, which enhanced the seed germination, root length, shoot length, and dry weight of chick pea, pigeon pea, and ground nut plants under pot studies. The efficient growth of the plants was not only due to the biocontrol activity of the siderophore produced by P. aeruginosa JAS-25 but also may be by the production of indole acetic acid (IAA), which influences the growth of the plants as phytohormones.

  19. C-di-GMP regulates antimicrobial peptide resistance in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Chua, Song Lin; Tan, Sean Yang-Yi; Rybtke, Morten Theil

    2013-01-01

    Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is an intracellular second messenger which controls the life styles of many bacteria. A high intracellular level of c-di-GMP induces a biofilm lifestyle, whereas a low intracellular level of c-di-GMP stimulates dispersal of biofilms and promotes a plankto......Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is an intracellular second messenger which controls the life styles of many bacteria. A high intracellular level of c-di-GMP induces a biofilm lifestyle, whereas a low intracellular level of c-di-GMP stimulates dispersal of biofilms and promotes...... a planktonic lifestyle. Here, we used expression of different reporters to show that planktonic cells (PCells), biofilm cells (BCells) and cells dispersed from biofilms (DCells) had distinct intracellular c-di-GMP levels. Proteomics analysis showed that the low intracellular c-di-GMP level of DCells induced...... the expression of proteins required for the virulence and development of antimicrobial peptide resistance in P. aeruginosa. In accordance, P. aeruginosa cells with low c-di-GMP levels were found to be more resistant to colistin than P. aeruginosa cells with high c-di-GMP levels. This contradicts the current...

  20. The immune system vs. Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Jensen, Peter Østrup; Givskov, Michael; Bjarnsholt, Thomas

    2010-01-01

    revealed both innate as well as adaptive immune responses to biofilms. On the other hand, measures launched by biofilm bacteria to achieve protection against the various immune responses have also been demonstrated. Whether particular immune responses to biofilm infections exist remains to be firmly...... established. However, because biofilm infections are often persistent (or chronic), an odd situation appears with the simultaneous activation of both arms of the host immune response, neither of which can eliminate the biofilm pathogen, but instead, in synergy, causes collateral tissue damage. Although...... the present review on the immune system vs. biofilm bacteria is focused on Pseudomonas aeruginosa (mainly because this is the most thoroughly studied), many of the same mechanisms are also seen with biofilm infections generated by other microorganisms....

  1. Flagellum-Mediated Biofilm Defense Mechanisms of Pseudomonas aeruginosa against Host-Derived Lactoferrin ▿

    Science.gov (United States)

    Leid, Jeff G.; Kerr, Mathias; Selgado, Candice; Johnson, Chelsa; Moreno, Gabriel; Smith, Alyssa; Shirtliff, Mark E.; O'Toole, George A.; Cope, Emily K.

    2009-01-01

    Chronic infection with the gram-negative organism Pseudomonas aeruginosa is a leading cause of morbidity and mortality in human patients, despite high doses of antibiotics used to treat the various diseases this organism causes. These infections are chronic because P. aeruginosa readily forms biofilms, which are inherently resistant to antibiotics as well as the host's immune system. Our laboratory has been investigating specific mutations in P. aeruginosa that regulate biofilm bacterial susceptibility to the host. To continue our investigation of the role of genetics in bacterial biofilm host resistance, we examined P. aeruginosa biofilms that lack the flgK gene. This mutant lacks flagella, which results in defects in early biofilm development (up to 36 h). For these experiments, the flgK-disrupted strain and the parental strain (PA14) were used in a modified version of the 96-well plate microtiter assay. Biofilms were challenged with freshly isolated human leukocytes for 4 to 6 h and viable bacteria enumerated by CFU. Subsequent to the challenge, both mononuclear cells (monocytes and lymphocytes) and neutrophils, along with tumor necrosis factor alpha (TNF-α), were required for optimal killing of the flgK biofilm bacteria. We identified a cytokine cross talk network between mononuclear cells and neutrophils that was essential to the production of lactoferrin and bacterial killing. Our data suggest that TNF-α is secreted from mononuclear cells, causing neutrophil activation, resulting in the secretion of bactericidal concentrations of lactoferrin. These results extend previous studies of the importance of lactoferrin in the innate immune defense against bacterial biofilms. PMID:19651866

  2. Biofilm Formation Mechanisms of Pseudomonas aeruginosa Predicted via Genome-Scale Kinetic Models of Bacterial Metabolism.

    Science.gov (United States)

    Vital-Lopez, Francisco G; Reifman, Jaques; Wallqvist, Anders

    2015-10-01

    A hallmark of Pseudomonas aeruginosa is its ability to establish biofilm-based infections that are difficult to eradicate. Biofilms are less susceptible to host inflammatory and immune responses and have higher antibiotic tolerance than free-living planktonic cells. Developing treatments against biofilms requires an understanding of bacterial biofilm-specific physiological traits. Research efforts have started to elucidate the intricate mechanisms underlying biofilm development. However, many aspects of these mechanisms are still poorly understood. Here, we addressed questions regarding biofilm metabolism using a genome-scale kinetic model of the P. aeruginosa metabolic network and gene expression profiles. Specifically, we computed metabolite concentration differences between known mutants with altered biofilm formation and the wild-type strain to predict drug targets against P. aeruginosa biofilms. We also simulated the altered metabolism driven by gene expression changes between biofilm and stationary growth-phase planktonic cultures. Our analysis suggests that the synthesis of important biofilm-related molecules, such as the quorum-sensing molecule Pseudomonas quinolone signal and the exopolysaccharide Psl, is regulated not only through the expression of genes in their own synthesis pathway, but also through the biofilm-specific expression of genes in pathways competing for precursors to these molecules. Finally, we investigated why mutants defective in anthranilate degradation have an impaired ability to form biofilms. Alternative to a previous hypothesis that this biofilm reduction is caused by a decrease in energy production, we proposed that the dysregulation of the synthesis of secondary metabolites derived from anthranilate and chorismate is what impaired the biofilms of these mutants. Notably, these insights generated through our kinetic model-based approach are not accessible from previous constraint-based model analyses of P. aeruginosa biofilm

  3. Ambroxol interferes with Pseudomonas aeruginosa quorum sensing.

    Science.gov (United States)

    Lu, Qi; Yu, Jialin; Yang, Xiqiang; Wang, Jiarong; Wang, Lijia; Lin, Yayin; Lin, Lihua

    2010-09-01

    The mucolytic agent ambroxol has been reported to interfere with the formation of Pseudomonas aeruginosa-derived biofilms in addition to reducing alginate production by undefined mechanisms. Since quorum sensing is a key regulator of virulence and biofilm formation, we examined the effects of ambroxol on P. aeruginosa PAO1 wild-type bacterial clearance rates, adhesion profiles and biofilm formation compared with the quorum sensing-deficient, double-mutant strains DeltalasR DeltarhlR and DeltalasI DeltarhlI. Data presented in this report demonstrated that ambroxol treatment reduced survival rates of the double-mutant strains compared with the wild-type strain in a dose-dependent manner even though the double-mutants had increased adhesion in the presence of ambroxol compared with the wild-type strain. The PAO1 wild-type strain produced a significantly thicker biofilm (21.64+/-0.57 microm) compared with the biofilms produced by the DeltalasR DeltarhlR (7.36+/-0.2 microm) and DeltalasI DeltarhlI (6.62+/-0.31 microm) isolates. Ambroxol treatment reduced biofilm thickness, increased areal porosity, and decreased the average diffusion distance and textual entropy of wild-type and double-mutant strains. However, compared with the double-mutant strains, the changes observed for the wild-type strain were more clearly defined. Finally, ambroxol exhibited significant antagonistic quorum-sensing properties, suggesting that it could be adapted for use clinically in the treatment of cystic fibrosis and to reduce biofilm formation and in the colonisation of indwelling devices.

  4. Alginate overproduction affects Pseudomonas aeruginosa biofilm structure and function

    DEFF Research Database (Denmark)

    Hentzer, Morten; Teitzel, G.M.; Balzer, G.J.;

    2001-01-01

    During the course of chronic cystic fibrosis (CF) infections, Pseudomonas aeruginosa undergoes a conversion to a mucoid phenotype, which is characterized by overproduction of the exopolysaccharide alginate. Chronic P. aeruginosa infections involve surface-attached, highly antibiotic-resistant com......During the course of chronic cystic fibrosis (CF) infections, Pseudomonas aeruginosa undergoes a conversion to a mucoid phenotype, which is characterized by overproduction of the exopolysaccharide alginate. Chronic P. aeruginosa infections involve surface-attached, highly antibiotic......-resistant communities of microorganisms organized in biofilms. Although biofilm formation and the conversion to mucoidy are both important aspects of CF pathogenesis, the relationship between them is at the present unclear. In this study, we report that the overproduction of alginate affects biofilm development...

  5. Oxylipins produced by Pseudomonas aeruginosa promote biofilm formation and virulence

    Science.gov (United States)

    Martínez, Eriel; Campos-Gómez, Javier

    2016-01-01

    The oxygenation of unsaturated fatty acids by dioxygenases occurs in all kingdoms of life and produces physiologically important lipids called oxylipins. The biological roles of oxylipins have been extensively studied in animals, plants, algae and fungi, but remain largely unidentified in prokaryotes. The bacterium Pseudomonas aeruginosa displays a diol synthase activity that transforms several monounsaturated fatty acids into mono- and di-hydroxylated derivatives. Here we show that oxylipins derived from this activity inhibit flagellum-driven motility and upregulate type IV pilus-dependent twitching motility of P. aeruginosa. Consequently, these oxylipins promote bacterial organization in microcolonies, increasing the ability of P. aeruginosa to form biofilms in vitro and in vivo (in Drosophila flies). We also demonstrate that oxylipins produced by P. aeruginosa promote virulence in Drosophila flies and lettuce. Our study thus uncovers a role for prokaryotic oxylipins in the physiology and pathogenicity of bacteria. PMID:27929111

  6. Acquisition and role of molybdate in Pseudomonas aeruginosa.

    Science.gov (United States)

    Pederick, Victoria G; Eijkelkamp, Bart A; Ween, Miranda P; Begg, Stephanie L; Paton, James C; McDevitt, Christopher A

    2014-11-01

    In microaerophilic or anaerobic environments, Pseudomonas aeruginosa utilizes nitrate reduction for energy production, a process dependent on the availability of the oxyanionic form of molybdenum, molybdate (MoO4 (2-)). Here, we show that molybdate acquisition in P. aeruginosa occurs via a high-affinity ATP-binding cassette permease (ModABC). ModA is a cluster D-III solute binding protein capable of interacting with molybdate or tungstate oxyanions. Deletion of the modA gene reduces cellular molybdate concentrations and results in inhibition of anaerobic growth and nitrate reduction. Further, we show that conditions that permit nitrate reduction also cause inhibition of biofilm formation and an alteration in fatty acid composition of P. aeruginosa. Collectively, these data highlight the importance of molybdate for anaerobic growth of P. aeruginosa and reveal novel consequences of nitrate reduction on biofilm formation and cell membrane composition.

  7. The Enzymes of the Ammonia Assimilation in Pseudomonas aeruginosa

    NARCIS (Netherlands)

    Janssen, Dick B.; Camp, Huub J.M. op den; Leenen, Pieter J.M.; Drift, Chris van der

    1980-01-01

    Glutamine synthetase from Pseudomonas aeruginosa is regulated by repression/derepression of enzyme synthesis and by adenylylation/deadenylylation control. High levels of deadenylylated biosynthetically active glutamine synthetase were observed in cultures growing with limiting amounts of nitrogen wh

  8. Vaccines for preventing infection with Pseudomonas aeruginosa in cystic fibrosis

    DEFF Research Database (Denmark)

    Johansen, H.K.; Gøtzsche, Peter C.; Johansen, Helle Krogh

    2008-01-01

    BACKGROUND: Chronic pulmonary infection in cystic fibrosis results in progressive lung damage. Once colonisation of the lungs with Pseudomonas aeruginosa occurs, it is almost impossible to eradicate. Vaccines, aimed at reducing infection with Pseudomonas aeruginosa, have been developed. OBJECTIVES......: To assess the effectiveness of vaccination against Pseudomonas aeruginosa in cystic fibrosis. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register using the terms vaccines AND pseudomonas (last search May 2008) and PubMed using the terms vaccin* AND cystic...... fibrosis (last search May 2008). SELECTION CRITERIA: Randomised trials (published or unpublished) comparing Pseudomonas aeruginosa vaccines (oral, parenteral or intranasal) with control vaccines or no intervention in cystic fibrosis. DATA COLLECTION AND ANALYSIS: The authors independently selected trials...

  9. Caenorhabditis elegans reveals novel Pseudomonas aeruginosa virulence mechanism

    NARCIS (Netherlands)

    Utari, Putri Dwi; Quax, Wim J.

    2013-01-01

    The susceptibility of Caenorhabditis elegans to different virulent phenotypes of Pseudomonas aeruginosa makes the worms an excellent model for studying host-pathogen interactions. Including the recently described liquid killing, five different killing assays are now available offering superb possibi

  10. Isolation of clinical strains of Pseudomonas aeruginosa harboring different plasmids.

    Science.gov (United States)

    Ranjbar, R; Owlia, P; Saderi, H; Bameri, Z; Izadi, M; Jonaidi, N; Morovvati, S

    2007-09-01

    Aim of this study was to investigate the presence of plasmids among the strains of P. aeruginosa isolated from clinically diagnosed cases in Tehran in 2006. A total of 38 strains of P. aeruginosa were isolated. With the exception of one isolate, all P. aeruginosa strains harbored at least one plasmid band. The electrophoretic analysis of plasmid DNAs showed different number of plasmid bands among the strains tested. The DNA band of 1.4 kbp was evident in 84.2% of the strains. Approximately 71 and 21% of the isolates harbored concomitantly two and three plasmids, respectively. Isolation of strains with diverse types of plasmids suggests the different cluster of P. aeruginosa might be disseminated during the current study period.

  11. Enhanced degradation of eletrooxidized textile effluent by petroleum degrading Pseudomonas aeruginosa (MTCC No.1201) at compressed gas pressure.

    Science.gov (United States)

    Santhanam, Manikandan; Annamalai, Sivasankar; Umarkatha, Sayera Banu; Sundaram, Maruthamuthu

    2015-03-01

    The textile dyeing industry produces large volumes of wastewater during dyeing processes where the major step includes the color removal and COD removal. In the present study, the combined electrooxidation process and a novel biological degradation at high compressed gas pressure were studied. The removal of color in the real textile dye effluent was achieved by electrooxidation with Titanium Substrate Insoluble anode and titanium as cathode through generation of hypochlorite. The hypochlorite produced during the electrooxidation was removed by exposing the solution to direct sunlight. The impact of compressed atmospheric condition on the degradation of organics by Pseudomonas aeruginosa (MTCC No.1201, GenBank Acc. No KC545414) was studied. The compressed gas pressure condition increases the level of dissolved gas in the liquid phase and exerts the pressure on the growing cells in the liquid phase. Interesting synchronization between the utilization of oxygen by active microbial cells and the dissolution of oxygen in the water from gas phase was observed which enhanced the bacterial degradation process. It should be mentioned here that the P. aeruginosa was grown without addition of nutrients. The compressed atmospheric pressure enhances the bacterial proliferation, EPS production and COD reduction in the electrooxidized effluent. FTIR and HPLC reveal the degradation of organics in the compressed pressure condition.

  12. Extracellular DNA Shields against Aminoglycosides in Pseudomonas aeruginosa Biofilms

    DEFF Research Database (Denmark)

    Chiang, Wen-Chi; Nilsson, Martin; Jensen, Peter Østrup

    2013-01-01

    , which are thought to be a source of extracellular DNA at sites of infections, increases the tolerance of P. aeruginosa biofilms toward aminoglycosides. Although biofilm-associated aminoglycoside tolerance recently has been linked to extracellular DNA-mediated activation of the pmr genes, we demonstrate...... that the aminoglycoside tolerance mediated by the presence of extracellular DNA is not caused by activation of the pmr genes in our P. aeruginosa biofilms but rather by a protective shield effect of the extracellular DNA....

  13. Genome Sequence of the Urethral Isolate Pseudomonas aeruginosa RN21

    OpenAIRE

    Wibberg, Daniel; Tielen, Petra; Narten, Maike; Schobert, Max; Blom, Jochen; Schatschneider, Sarah; Meyer, Ann-Kathrin; Neubauer, Rüdiger; Albersmeier, Andreas; Albaum, Stefan; Jahn, Martina; Goesmann, Alexander; Vorhölter, Frank-Jörg; Pühler, Alfred; Jahn, Dieter

    2015-01-01

    Pseudomonas aeruginosa is known to cause complicated urinary tract infections (UTI). The improved 7.0-Mb draft genome sequence of P. aeruginosa RN21, isolated from a patient with an acute UTI, was determined. It carries three (pro)phage genomes, genes for two restriction/modification systems, and a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system.

  14. Membrane-bound respiratory chain of Pseudomonas aeruginosa grown aerobically.

    OpenAIRE

    Matsushita, K.; Yamada, M.; Shinagawa, E; Adachi, O; Ameyama, M

    1980-01-01

    The electron transport chain of the gram-negative bacterium Pseudomonas aeruginosa, grown aerobically, contained a number of primary dehydrogenases and respiratory components (soluble flavin, bound flavin, coenzyme Q9, heme b, heme c, and cytochrome o) in membrane particles of the organism. Cytochrome o, about 50% of the b-type cytochrome, seemed to function as a terminal oxidase in the respiratory chain. The electron transport chain of P. aeruginosa grown aerobically was suggested to be line...

  15. Pseudomonas aeruginosa PAO1 Kills Caenorhabditis elegans by Cyanide Poisoning

    OpenAIRE

    Gallagher, Larry A.; Manoil, Colin

    2001-01-01

    In this report we describe experiments to investigate a simple virulence model in which Pseudomonas aeruginosa PAO1 rapidly paralyzes and kills the nematode Caenorhabditis elegans. Our results imply that hydrogen cyanide is the sole or primary toxic factor produced by P. aeruginosa that is responsible for killing of the nematode. Four lines of evidence support this conclusion. First, a transposon insertion mutation in a gene encoding a subunit of hydrogen cyanide synthase (hcnC) eliminated ne...

  16. Singly Flagellated Pseudomonas aeruginosa Chemotaxes Efficiently by Unbiased Motor Regulation

    Directory of Open Access Journals (Sweden)

    Qiuxian Cai

    2016-04-01

    Full Text Available Pseudomonas aeruginosa is an opportunistic human pathogen that has long been known to chemotax. More recently, it has been established that chemotaxis is an important factor in the ability of P. aeruginosa to make biofilms. Genes that allow P. aeruginosa to chemotax are homologous with genes in the paradigmatic model organism for chemotaxis, Escherichia coli. However, P. aeruginosa is singly flagellated and E. coli has multiple flagella. Therefore, the regulation of counterclockwise/clockwise flagellar motor bias that allows E. coli to efficiently chemotax by runs and tumbles would lead to inefficient chemotaxis by P. aeruginosa, as half of a randomly oriented population would respond to a chemoattractant gradient in the wrong sense. How P. aeruginosa regulates flagellar rotation to achieve chemotaxis is not known. Here, we analyze the swimming trajectories of single cells in microfluidic channels and the rotations of cells tethered by their flagella to the surface of a variable-environment flow cell. We show that P. aeruginosa chemotaxes by symmetrically increasing the durations of both counterclockwise and clockwise flagellar rotations when swimming up the chemoattractant gradient and symmetrically decreasing rotation durations when swimming down the chemoattractant gradient. Unlike the case for E. coli, the counterclockwise/clockwise bias stays constant for P. aeruginosa. We describe P. aeruginosa’s chemotaxis using an analytical model for symmetric motor regulation. We use this model to do simulations that show that, given P. aeruginosa’s physiological constraints on motility, its distinct, symmetric regulation of motor switching optimizes chemotaxis.

  17. Laser removal of tattoos.

    Science.gov (United States)

    Kuperman-Beade, M; Levine, V J; Ashinoff, R

    2001-01-01

    Tattoos are placed for different reasons. A technique for tattoo removal which produces selective removal of each tattoo pigment, with minimal risk of scarring, is needed. Nonspecific methods have a high incidence of scarring, textural, and pigmentary alterations compared with the use of Q-switched lasers. With new advances in Q-switched laser technology, tattoo removal can be achieved with minimal risk of scarring and permanent pigmentary alteration. There are five types of tattoos: amateur, professional, cosmetic, medicinal, and traumatic. Amateur tattoos require less treatment sessions than professional multicolored tattoos. Other factors to consider when evaluating tattoos for removal are: location, age and the skin type of the patient. Treatment should begin by obtaining a pre-operative history. Since treatment with the Q-switched lasers is painful, use of a local injection with lidocaine or topical anaesthesia cream may be used prior to laser treatment. Topical broad-spectrum antibacterial ointment is applied immediately following the procedure. Three types of lasers are currently used for tattoo removal: Q-switched ruby laser (694 nm), Q-switched Nd:YAG laser (532 nm, 1064 nm), and Q-switched alexandrite laser (755 nm). The Q-switched ruby and alexandrite lasers are useful for removing black, blue and green pigments. The Q-switched 532 nm Nd:YAG laser can be used to remove red pigments and the 1064 nm Nd:YAG laser is used for removal of black and blue pigments. The most common adverse effects following laser tattoo treatment with the Q-switched ruby laser include textural change, scarring, and pigmentary alteration. Transient hypopigmentation and textural changes have been reported in up to 50 and 12%, respectively, of patients treated with the Q-switched alexandrite laser. Hyperpigmentation and textural changes are infrequent adverse effects of the Q-switched Nd:YAG laser and the incidence of hypopigmentary changes is much lower than with the ruby laser

  18. Polymicrobial Ventilator-Associated Pneumonia: Fighting In Vitro Candida albicans-Pseudomonas aeruginosa Biofilms with Antifungal-Antibacterial Combination Therapy

    Science.gov (United States)

    Pereira, Cláudia R.; Azevedo, Nuno F.; Lourenço, Anália; Henriques, Mariana; Pereira, Maria O.

    2017-01-01

    The polymicrobial nature of ventilator-associated pneumonia (VAP) is now evident, with mixed bacterial-fungal biofilms colonizing the VAP endotracheal tube (ETT) surface. The microbial interplay within this infection may contribute for enhanced pathogenesis and exert impact towards antimicrobial therapy. Consequently, the high mortality/morbidity rates associated to VAP and the worldwide increase in antibiotic resistance has promoted the search for novel therapeutic strategies to fight VAP polymicrobial infections. Under this scope, this work aimed to assess the activity of mono- vs combinational antimicrobial therapy using one antibiotic (Polymyxin B; PolyB) and one antifungal (Amphotericin B; AmB) agent against polymicrobial biofilms of Pseudomonas aeruginosa and Candida albicans. The action of isolated antimicrobials was firstly evaluated in single- and polymicrobial cultures, with AmB being more effective against C. albicans and PolyB against P. aeruginosa. Mixed planktonic cultures required equal or higher antimicrobial concentrations. In biofilms, only PolyB at relatively high concentrations could reduce P. aeruginosa in both monospecies and polymicrobial populations, with C. albicans displaying only punctual disturbances. PolyB and AmB exhibited a synergistic effect against P. aeruginosa and C. albicans mixed planktonic cultures, but only high doses (256 mg L-1) of PolyB were able to eradicate polymicrobial biofilms, with P. aeruginosa showing loss of cultivability (but not viability) at 2 h post-treatment, whilst C. albicans only started to be inhibited after 14 h. In conclusion, combination therapy involving an antibiotic and an antifungal agent holds an attractive therapeutic option to treat severe bacterial-fungal polymicrobial infections. Nevertheless, optimization of antimicrobial doses and further clinical pharmacokinetics/pharmacodynamics and toxicodynamics studies underpinning the optimal use of these drugs are urgently required to improve therapy

  19. Selective eicosanoid-generating capacity of cytoplasmic phospholipase A2 in Pseudomonas aeruginosa-infected epithelial cells.

    Science.gov (United States)

    Hurley, Bryan P; Pirzai, Waheed; Mumy, Karen L; Gronert, Karsten; McCormick, Beth A

    2011-02-01

    Airway neutrophil infiltration is a pathological hallmark observed in multiple lung diseases including pneumonia and cystic fibrosis. Bacterial pathogens such as Pseudomonas aeruginosa instigate neutrophil recruitment to the air space. Excessive accumulation of neutrophils in the lung often contributes to tissue destruction. Previous studies have unveiled hepoxilin A(3) as the key molecular signal driving neutrophils across epithelial barriers. The eicosanoid hepoxilin A(3) is a potent neutrophil chemoattractant produced by epithelial cells in response to infection with P. aeruginosa. The enzyme phospholipase A(2) liberates arachidonic acid from membrane phospholipids, the rate-limiting step in the synthesis of all eicosanoids, including hepoxilin A(3). Once generated, aracidonic acid is acted upon by multiple cyclooxygenases and lipoxygenases producing an array of functionally diverse eicosanoids. Although there are numerous phospholipase A(2) isoforms capable of generating arachidonic acid, the isoform most often associated with eicosanoid generation is cytoplasmic phospholipase A(2)α. In the current study, we observed that the cytoplasmic phospholipase A(2)α isoform is required for mediating P. aeruginosa-induced production of certain eicosanoids such as prostaglandin E(2). However, we found that neutrophil transepithelial migration induced by P. aeruginosa does not require cytoplasmic phospholipase A(2)α. Furthermore, P. aeruginosa-induced hepoxilin A(3) production persists despite cytoplasmic phospholipase A(2)α suppression and generation of the 12-lipoxygenase metabolite 12-HETE is actually enhanced in this context. These results suggest that alterative phospholipase A(2) isoforms are utilized to synthesize 12-lipoxygenase metabolites. The therapeutic implications of these findings are significant when considering anti-inflammatory therapies based on targeting eicosanoid synthesis pathways.

  20. Antimicrobial efficacy of silver ions in combination with tea tree oil against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans.

    Science.gov (United States)

    Low, W L; Martin, C; Hill, D J; Kenward, M A

    2011-02-01

    Tea tree oil (TTO) and silver ions (Ag(+)), either alone or in combination with other antimicrobial compounds, have been used in the treatment of topical infections. However, there appears to be little data on the efficacy of TTO combined with silver in the absence of any other agents. TTO and Ag(+) were added, alone and in combination, to suspension cultures of Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. Treatment of these cultures with TTO and Ag(+) at sub-minimal lethal concentrations resulted in an enhanced loss of viability compared with treatment with individual agents. The order of sensitivity to the combined agents was P. aeruginosa>S. aureus>C. albicans. The fractional lethal concentration index (FLCI) showed that these combinations of TTO and Ag(+) exerted a synergistic effect against P. aeruginosa (FLCI=0.263) and an indifferent effect against S. aureus and C. albicans (FLCI=0.663 and 1.197, respectively). The results indicate that combining these antimicrobial agents may be useful in decreasing the concentration of antimicrobial agents required to achieve an effective reduction in opportunistic pathogenic microorganisms that typically infect wounds.

  1. Resistant patterns of Pseudomonas aeruginosa in a Malaysian teaching hospital

    Institute of Scientific and Technical Information of China (English)

    Zaidah AR; Siti SMN; Zahiruddin WM; Zeehaida M

    2009-01-01

    Objective:Pseudomonas aeruginosa is an opportunistic pathogen and the leading cause of nosocomial infec-tions.Currently a notable increase in the prevalence of multidrug-resistant P.aeruginosa worldwide has been reported in hospitalized patients and was associated with high morbidity and mortality.Methods:A retrospec-tive laboratory based analysis regarding the spectrum and distribution of P.aeruginosa from a wide range of clinical samples in Hospital Universiti Sains Malaysia since January 2003 to December 2007 was done.Re-sults:Altogether,there were 2 308 clinical isolates analyzed.The main sources of P.aeruginosa were from swab,respiratory,urine and blood specimens which accounted for 28.2 %,21.8 %,13.2 % and 12.8 %respectively.Results showed significant reduction in percentage of resistant towards three antibiotic namely ciprofloxacin,ceftazidime and imipenem.However the percentage of pan-resistant P.aeruginosa increased steadily over these years.Conclusion:This data is helpful to the clinician in guiding the choice of appropriate antibiotic to treat P.aeruginosa infection.At the same time,it warrants a more aggressive infection control ac-tivity to be implemented to control the spread of pan resistant strain in this centre.

  2. Inhibitory effect of zinc oxide nanoparticles on pseudomonas aeruginosa biofilm formation

    Directory of Open Access Journals (Sweden)

    Mohammad Hassani Sangani

    2015-04-01

    Full Text Available Objective(s: Bacterial biofilm formation causes many persistent and chronic infections. The matrix protects biofilm bacteria from exposure to innate immune defenses and antibiotic treatments. The purpose of this study was to evaluate the biofilm formation of clinical isolates of Pseudomonas aeruginosa and the activity of zinc oxide nanoparticles (ZnO NPs on biofilm. Materials and Methods: After collecting bacteria from clinical samples of hospitalized patients, the ability of organisms were evaluated to create biofilm by tissue culture plate (TCP assay. ZnO NPs were synthesized by sol gel method and the efficacy of different concentrations (50- 350 µg/ml of ZnO NPs was assessed on biofilm formation and also elimination of pre-formed biofilm by using TCP method. Results:The average diameter of synthesized ZnO NPs was 20 nm. The minimum inhibitory concentration of nanoparticles was 150- 158 μg/ml and the minimum bactericidal concentration was higher (325 µg/ml. All 15 clinical isolates of P. aeruginosa were able to produce biofilm. Treating the organisms with nanoparticles at concentrations of 350 μg/ml resulted in more than 94% inhibition in OD reduction%. Molecular analysis showed that the presence of mRNA of pslA gene after treating bacteria with ZnO NPs for 30 minutes. Conclusion: The results showed that ZnO NPs can inhibit the establishment of P. aeruginosa biofilms and have less effective in removing pre-formed biofilm. However the tested nanoparticles exhibited anti-biofilm effect, but mRNA of pslA gene could be still detected in the medium by RT-PCR technique after 30 minutes treatment with ZnO.

  3. Enzymes Enhance Biofilm Removal Efficiency of Cleaners.

    Science.gov (United States)

    Stiefel, Philipp; Mauerhofer, Stefan; Schneider, Jana; Maniura-Weber, Katharina; Rosenberg, Urs; Ren, Qun

    2016-06-01

    Efficient removal of biofilms from medical devices is a big challenge in health care to avoid hospital-acquired infections, especially from delicate devices like flexible endoscopes, which cannot be reprocessed using harsh chemicals or high temperatures. Therefore, milder solutions such as enzymatic cleaners have to be used, which need to be carefully developed to ensure efficacious performance. In vitro biofilm in a 96-well-plate system was used to select and optimize the formulation of novel enzymatic cleaners. Removal of the biofilm was quantified by crystal violet staining, while the disinfecting properties were evaluated by a BacTiter-Glo assay. The biofilm removal efficacy of the selected cleaner was further tested by using European standard (EN) for endoscope cleaning EN ISO 15883, and removal of artificial blood soil was investigated by treating TOSI (Test Object Surgical Instrument) cleaning indicators. Using the process described here, a novel enzymatic endoscope cleaner was developed, which removed 95% of Staphylococcus aureus and 90% of Pseudomonas aeruginosa biofilms in the 96-well plate system. With a >99% reduction of CFU and a >90% reduction of extracellular polymeric substances, this cleaner enabled subsequent complete disinfection and fulfilled acceptance criteria of EN ISO 15883. Furthermore, it efficiently removed blood soil and significantly outperformed comparable commercial products. The cleaning performance was stable even after storage of the cleaner for 6 months. It was demonstrated that incorporation of appropriate enzymes into the cleaner enhanced performance significantly.

  4. Advanced Coating Removal Techniques

    Science.gov (United States)

    Seibert, Jon

    2006-01-01

    An important step in the repair and protection against corrosion damage is the safe removal of the oxidation and protective coatings without further damaging the integrity of the substrate. Two such methods that are proving to be safe and effective in this task are liquid nitrogen and laser removal operations. Laser technology used for the removal of protective coatings is currently being researched and implemented in various areas of the aerospace industry. Delivering thousands of focused energy pulses, the laser ablates the coating surface by heating and dissolving the material applied to the substrate. The metal substrate will reflect the laser and redirect the energy to any remaining protective coating, thus preventing any collateral damage the substrate may suffer throughout the process. Liquid nitrogen jets are comparable to blasting with an ultra high-pressure water jet but without the residual liquid that requires collection and removal .As the liquid nitrogen reaches the surface it is transformed into gaseous nitrogen and reenters the atmosphere without any contamination to surrounding hardware. These innovative technologies simplify corrosion repair by eliminating hazardous chemicals and repetitive manual labor from the coating removal process. One very significant advantage is the reduction of particulate contamination exposure to personnel. With the removal of coatings adjacent to sensitive flight hardware, a benefit of each technique for the space program is that no contamination such as beads, water, or sanding residue is left behind when the job is finished. One primary concern is the safe removal of coatings from thin aluminum honeycomb face sheet. NASA recently conducted thermal testing on liquid nitrogen systems and found that no damage occurred on 1/6", aluminum substrates. Wright Patterson Air Force Base in conjunction with Boeing and NASA is currently testing the laser remOval technique for process qualification. Other applications of liquid

  5. The Study of Synergistic Effects of n.butanolic Cyclamen coum Extract and Ciprofloxacin on inhibition of Pseudomonas aeruginosa biofilm formation

    Directory of Open Access Journals (Sweden)

    ahya abdi ali

    2015-02-01

    Full Text Available   Introduction : Infections caused by Pseudomonas aeruginosa biofilm are the major causes of death in patients with cystic fibrosis (CF. Some studies revealed that biofilms are resistant to several antibiotics because of their impermeable structures. In order to re-sensitize bacteria to different antibiotics, biofilm formation should be inhibited. In this research, evaluation of antibiofilm activity of n-butanolic Cyclamen coum extract as a medici­nal plant from Myrsinaceae family, in combination with ciprofloxacin was carried out.   Materials and method s: The biofilm formation ability by P. aeruginosa PAO1 and one clinically isolated P. aeruginosa (PA214 was confirmed by microtiter plate method. Extraction of the tubers of Cyclamen coum was done by fractionation method . The antibiofilm and antibacterial properties of n-butanolic C. coum extract (which includes saponin compounds alone and in combination with ciprofloxacin by using microdilution and crystal violet methods were examined. The cytotoxicity effect of the n-butanolic extract on HT-29 cells was assayed by MTT (3- (4,5-dimethylthiazol-2-yl -2,5-diphenyl-tetrazolium bromide test.   Results : The biofilm formation ability by P. aeruginosa strains was quantitatively confirmed. Saponin content of the n-butanolic C.coum extract was 156 µg/mL. The extract revealed antibacterial activity against the growth of planktonic P. aeruginosa strains. The combination of n-butanolic C.coum extract and ciprofloxacin significantly inhibited P.aeruginosa biofilm formation (ΣFBIC = 0.5. The n-butanolic C.coum extract showed insignificant cytotoxic effect against HT-29 human cancer cell line after 48 hours and 72 hours incubation .   Discussion and conclusion : It can be concluded that n-butanolic C.coum extract in combination with ciprofloxacin significantly revealed antibiofilm activity against P. aeruginosa biofilm however, further clinical investigations are required.

  6. North Korea: Terrorism List Removal?

    Science.gov (United States)

    2008-07-10

    calender -day notification period to Congress as required by U.S. law. The White House stated that North Korea would thus be removed on August 11, 2008...Congress notification of his intent to remove North Korea from the list of state sponsors of terrorism after 45 calender days. Under U.S. law, the

  7. North Korea: Terrorism List Removal

    Science.gov (United States)

    2008-11-06

    he was officially notifying Congress of his intent to remove North Korea from the list of state sponsors of terrorism after the 45 calender -day...of his intent to remove North Korea from the list of state sponsors of terrorism after 45 calender days. Under U.S. law, the President is required to

  8. Utility of lytic bacteriophage in the treatment of multidrug-resistant Pseudomonas aeruginosa septicemia in mice

    Directory of Open Access Journals (Sweden)

    Vinodkumar C

    2008-07-01

    Full Text Available Drug resistance is the major cause of increase in morbidity and mortality in neonates. One thousand six hundred forty-seven suspected septicemic neonates were subjected for microbiological analysis over a period of 5 years. Forty-two P. aeruginosa were isolated and the antibiogram revealed that 28 P. aeruginosa were resistant to almost all the common drugs used (multidrug-resistant. The emergence of antibiotic-resistant bacterial strains is one of the most critical problems of modern medicine. As a result, a novel and most effective approaches for treating infection caused by multidrug-resistant bacteria are urgently required. In this context, one intriguing approach is to use bacteriophages (viruses that kill bacteria in the treatment of infection caused by drug-resistant bacteria. In the present study, the utility of lytic bacteriophages to rescue septicemic mice with multidrug-resistant (MDR P. aeruginosa infection was evaluated. MDR P. aeruginosa was used to induce septicemia in mice by intraperitoneal (i.p. injection of 10 7 CFU. The resulting bacteremia was fatal within 48 hrs. The phage strain used in this study had lytic activity against a wide range of clinical isolates of MDR P. aeruginosa. A single i.p. injection of 3 x 10 9 PFU of the phage strain, administered 45 min after the bacterial challenge, was sufficient to rescue 100% of the animals. Even when treatment was delayed to the point where all animals were moribund, approximately 50% of them were rescued by a single injection of this phage preparation. The ability of this phage to rescue septicemic mice was demonstrated to be due to the functional capabilities of the phage and not to a nonspecific immune effect. The rescue of septicemic mice could be affected only by phage strains able to grow in vitro on the bacterial host used to infect the animals and when such strains are heat-inactivated, they lose their ability to rescue the infected mice. Multidrug-resistant bacteria have

  9. Pseudomonas aeruginosa Pore-Forming Exolysin and Type IV Pili Cooperate To Induce Host Cell Lysis

    Science.gov (United States)

    Basso, Pauline; Ragno, Michel; Elsen, Sylvie; Reboud, Emeline; Golovkine, Guillaume; Bouillot, Stephanie; Huber, Philippe; Lory, Stephen; Faudry, Eric

    2017-01-01

    ABSTRACT   Clinical strains of Pseudomonas aeruginosa lacking the type III secretion system genes employ a toxin, exolysin (ExlA), for host cell membrane disruption. Here, we demonstrated that ExlA export requires a predicted outer membrane protein, ExlB, showing that ExlA and ExlB define a new active two-partner secretion (TPS) system of P. aeruginosa. In addition to the TPS signals, ExlA harbors several distinct domains, which include one hemagglutinin domain, five arginine-glycine-aspartic acid (RGD) motifs, and a C-terminal region lacking any identifiable sequence motifs. However, this C-terminal region is important for the toxic activity, since its deletion abolishes host cell lysis. Using lipid vesicles and eukaryotic cells, including red blood cells, we demonstrated that ExlA has a pore-forming activity which precedes cell membrane disruption of nucleated cells. Finally, we developed a high-throughput cell-based live-dead assay and used it to screen a transposon mutant library of an ExlA-producing P. aeruginosa clinical strain for bacterial factors required for ExlA-mediated toxicity. The screen resulted in the identification of proteins involved in the formation of type IV pili as being required for ExlA to exert its cytotoxic activity by promoting close contact between bacteria and the host cell. These findings represent the first example of cooperation between a pore-forming toxin of the TPS family and surface appendages in host cell intoxication. PMID:28119472

  10. Nicotinamide nucleotide transhydrogenase (Nnt) links the substrate requirement in brain mitochondria for hydrogen peroxide removal to the thioredoxin/peroxiredoxin (Trx/Prx) system.

    Science.gov (United States)

    Lopert, Pamela; Patel, Manisha

    2014-05-30

    Mitochondrial reactive oxygen species are implicated in the etiology of multiple neurodegenerative diseases, including Parkinson disease. Mitochondria are known to be net producers of ROS, but recently we have shown that brain mitochondria can consume mitochondrial hydrogen peroxide (H2O2) in a respiration-dependent manner predominantly by the thioredoxin/peroxiredoxin system. Here, we sought to determine the mechanism linking mitochondrial respiration with H2O2 catabolism in brain mitochondria and dopaminergic cells. We hypothesized that nicotinamide nucleotide transhydrogenase (Nnt), which utilizes the proton gradient to generate NADPH from NADH and NADP(+), provides the link between mitochondrial respiration and H2O2 detoxification through the thioredoxin/peroxiredoxin system. Pharmacological inhibition of Nnt in isolated brain mitochondria significantly decreased their ability to consume H2O2 in the presence, but not absence, of respiration substrates. Nnt inhibition in liver mitochondria, which do not require substrates to detoxify H2O2, had no effect. Pharmacological inhibition or lentiviral knockdown of Nnt in N27 dopaminergic cells (a) decreased H2O2 catabolism, (b) decreased NADPH and increased NADP(+) levels, and (c) decreased basal, spare, and maximal mitochondrial oxygen consumption rates. Nnt-deficient cells possessed higher levels of oxidized mitochondrial Prx, which rendered them more susceptible to steady-state increases in H2O2 and cell death following exposure to subtoxic levels of paraquat. These data implicate Nnt as the critical link between the metabolic and H2O2 antioxidant function in brain mitochondria and suggests Nnt as a potential therapeutic target to improve the redox balance in conditions of oxidative stress associated with neurodegenerative diseases.

  11. Rhamnolipid production by pseudomonas aeruginosa GIM 32 using different substrates including molasses distillery wastewater.

    Science.gov (United States)

    Li, An-hua; Xu, Mei-ying; Sun, Wei; Sun, Guo-ping

    2011-03-01

    A rhamnolipid production strain newly isolated from oil-contaminated soil was identified as Pseudomonas aeruginosa GIM32 by its morphology and 16S rDNA sequence analysis. The effect of carbon source and carbon to nitrogen (C/N) ratio on rhamnolipids production was investigated. Palm oil was favorable as a carbon source for rhamnolipid production. The maximum biomass and rhamnolipid concentration were 8.24 g/L and 30.4 g/L, respectively, with an optimization medium containing 50 g/L palm oil and 5 g/L sodium nitrate. Molasses distillery wastewater as an unconventional substrate for rhamnolipid production was investigated. It was found that 2.6 g/L of rhamnolipids was produced; this amount was higher than that of past reports using wastewater as a substrate. In addition, 44% of the chemical oxygen demand of wastewater was removed at the same time under the optimization condition. Eleven kinds of different molecular weight rhamnolipid homologues were identified in the rhamnolipids obtained from molasses distillery wastewater by P. aeruginosa GIM32 by LC-MS analysis.

  12. Biodegradation of Malathion using mixed culture of Serratia marcescens BNA1and Pseudomonas aeruginosa BNA2

    Directory of Open Access Journals (Sweden)

    B Nadalian

    2016-03-01

    Full Text Available Background and Objectives: Organophosphate pesticides are used most commonly for domestic, commercial, and agricultural purposes and have been found to be highly toxic. In essence, bioremediation has become one of the most important tools for removing these compounds in the environment, considering its higher efficiency when compared with the physicochemical methods. Materials and Methods: The biodegradation efficiency of two bacterial strains (i.e. Serratia marcescens BNA1 and Pseudomonas aeruginosa BNA2 were assessed. In order to evaluate Malathion biodegradation, each sample was cultured on mineral salts medium containing Malathion as a sole carbon source. Malathion biodegradation efficiency of the strains was monitored in different culture media. The ability of bacterial isolates to degrade Malathion was studied using gas chromatography. Results: Serratia marcescens BNA1 and Pseudomonas aeruginosa BNA2 were able to degrade Malathion. Biodegradation percentage in different treatments recorded were: BNA1+Ma (33.88%, BNA2+MA (26.45%, BNA1+BNA2+Ma (46/96%, BNA1+Ma+Tween (61.05%, BNA2+Ma+Tween (40.17%, and BNA1+BNA2+Ma+ Tween (67.79%. Conclusion: It could be speculated that the best degradation efficiency can be yielded using mixture of strains plus a surfactant. The results of this study can be used in the bioremediation of Malathion contaminate soil after doing the pilot experiments.

  13. Small Molecule Antivirulents Targeting the Iron-Regulated Heme Oxygenase (HemO) of P. aeruginosa

    OpenAIRE

    2013-01-01

    Bacteria require iron for survival and virulence and employ several mechanisms including utilization of the host heme containing proteins. The final step in releasing iron is the oxidative cleavage of heme by HemO. A recent computer aided drug design (CADD) study identified several inhibitors of the bacterial HemOs. Herein we report the near complete HN, N, CO Cα and Cβ chemical shift assignment of the P. aeruginosa HemO in the absence and presence of inhibitors (E)-3-(4-(Phenylamino)phenylca...

  14. Bioadsorption characteristics of Pseudomonas aeruginosa PAOI

    Directory of Open Access Journals (Sweden)

    Kőnig-Péter Anikó

    2014-01-01

    Full Text Available Biosorption of Cd(II and Pb(II ions from aqueous solution using lyophilized Pseudomonas aeruginosa (PAOI cells were observed under various experimental conditions. The effect of pH, initial metal concentration, equilibration time and temperature on bioadsorption was investigated. The optimum pH value for Pb(II adsorption was found to be 5.0, and for Cd(II 5.0 − 6.0. The Pb(II and Cd(II bioadsorption equilibrium were analyzed by using Freundlich and Langmuir model using nonlinear least-squares estimation. The experimental maximum uptake capacity of Pb(II and Cd(II was estimated to be 164 mg g-1 and 113 mg g-1, respectively. For biosorption kinetic study the pseudo second-order kinetic model was applied at various temperatures. The temperature had no significant effect on Pb(II bioadsorption. In case of Cd(II bioadsorption the adsorbed amount decreased with increasing temperature.

  15. Spaceflight promotes biofilm formation by Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Wooseong Kim

    Full Text Available Understanding the effects of spaceflight on microbial communities is crucial for the success of long-term, manned space missions. Surface-associated bacterial communities, known as biofilms, were abundant on the Mir space station and continue to be a challenge on the International Space Station. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of immune function observed during spaceflight. While planktonic cultures of microbes have indicated that spaceflight can lead to increases in growth and virulence, the effects of spaceflight on biofilm development and physiology remain unclear. To address this issue, Pseudomonas aeruginosa was cultured during two Space Shuttle Atlantis missions: STS-132 and STS-135, and the biofilms formed during spaceflight were characterized. Spaceflight was observed to increase the number of viable cells, biofilm biomass, and thickness relative to normal gravity controls. Moreover, the biofilms formed during spaceflight exhibited a column-and-canopy structure that has not been observed on Earth. The increase in the amount of biofilms and the formation of the novel architecture during spaceflight were observed to be independent of carbon source and phosphate concentrations in the media. However, flagella-driven motility was shown to be essential for the formation of this biofilm architecture during spaceflight. These findings represent the first evidence that spaceflight affects community-level behaviors of bacteria and highlight the importance of understanding how both harmful and beneficial human-microbe interactions may be altered during spaceflight.

  16. Genomic analysis and temperature-dependent transcriptome profiles of the rhizosphere originating strain Pseudomonas aeruginosa M18

    Directory of Open Access Journals (Sweden)

    He Ya-Wen

    2011-08-01

    Full Text Available Abstract Background Our previously published reports have described an effective biocontrol agent named Pseudomonas sp. M18 as its 16S rDNA sequence and several regulator genes share homologous sequences with those of P. aeruginosa, but there are several unusual phenotypic features. This study aims to explore its strain specific genomic features and gene expression patterns at different temperatures. Results The complete M18 genome is composed of a single chromosome of 6,327,754 base pairs containing 5684 open reading frames. Seven genomic islands, including two novel prophages and five specific non-phage islands were identified besides the conserved P. aeruginosa core genome. Each prophage contains a putative chitinase coding gene, and the prophage II contains a capB gene encoding a putative cold stress protein. The non-phage genomic islands contain genes responsible for pyoluteorin biosynthesis, environmental substance degradation and type I and III restriction-modification systems. Compared with other P. aeruginosa strains, the fewest number (3 of insertion sequences and the most number (3 of clustered regularly interspaced short palindromic repeats in M18 genome may contribute to the relative genome stability. Although the M18 genome is most closely related to that of P. aeruginosa strain LESB58, the strain M18 is more susceptible to several antimicrobial agents and easier to be erased in a mouse acute lung infection model than the strain LESB58. The whole M18 transcriptomic analysis indicated that 10.6% of the expressed genes are temperature-dependent, with 22 genes up-regulated at 28°C in three non-phage genomic islands and one prophage but none at 37°C. Conclusions The P. aeruginosa strain M18 has evolved its specific genomic structures and temperature dependent expression patterns to meet the requirement of its fitness and competitiveness under selective pressures imposed on the strain in rhizosphere niche.

  17. Phage ΦPan70, a Putative Temperate Phage, Controls Pseudomonas aeruginosa in Planktonic, Biofilm and Burn Mouse Model Assays

    Science.gov (United States)

    Holguín, Angela V.; Rangel, Guillermo; Clavijo, Viviana; Prada, Catalina; Mantilla, Marcela; Gomez, María Catalina; Kutter, Elizabeth; Taylor, Corinda; Fineran, Peter C.; Barrios, Andrés Fernando González; Vives, Martha J.

    2015-01-01

    Pseudomonas aeruginosa is one of the Multi-Drug-Resistant organisms most frequently isolated worldwide and, because of a shortage of new antibiotics, bacteriophages are considered an alternative for its treatment. Previously, P. aeruginosa phages were isolated and best candidates were chosen based on their ability to form clear plaques and their host range. This work aimed to characterize one of those phages, ΦPan70, preliminarily identified as a good candidate for phage-therapy. We performed infection curves, biofilm removal assays, transmission-electron-microscopy, pulsed-field-gel-electrophoresis, and studied the in vivo ΦPan70 biological activity in the burned mouse model. ΦPan70 was classified as a member of the Myoviridae family and, in both planktonic cells and biofilms, was responsible for a significant reduction in the bacterial population. The burned mouse model showed an animal survival between 80% and 100%, significantly different from the control animals (0%). However, analysis of the ΦPan70 genome revealed that it was 64% identical to F10, a temperate P. aeruginosa phage. Gene annotation indicated ΦPan70 as a new, but possible temperate phage, therefore not ideal for phage-therapy. Based on this, we recommend genome sequence analysis as an early step to select candidate phages for potential application in phage-therapy, before entering into a more intensive characterization. PMID:26274971

  18. Extraction of some strategic elements from thorium–uranium concentrate using bioproducts of Aspergillus ficuum and Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Osman A. Desouky

    2016-09-01

    Full Text Available The activity of the bioproducts from Aspergillus ficuum and Pseudomonas aeruginosa for extraction of thorium (Th4+, uranium (UO22+ and rare earth elements (REEs from thorium–uranium concentrate was studied. P. aeruginosa produce element-specific ligand (siderophore that is able to change pH and enhance chelation of Th4+ and UO22+. The produced siderophore at pH 5.3 has the ability to bioleach and is complexed with 68.00% of uranium and 65.00% of thorium. Also, A. ficuum produced different kinds of organic acids which leached 30.00% of uranium and 29.12% of thorium in addition to 20.00% of lanthanum, 33.00% of cerium and 2.51% of yttrium as rare earth elements at pH 3.0. Oxalic acid was efficient for Th4+, UO22+and REEs precipitation. The binocular stereo-microscope (BSM, environmental scanning electron microscope (ESEM and X-ray diffraction (XRD analyses confirmed the percentages of extracted metals. Exogenous polysaccharides (EPSs seem to play an important role in bioleaching and removal of these elements. It was found that EPSs produced by A. ficuum adsorbed Th4+, UO22+ and REEs while that produced by P. aeruginosa adsorbed REEs only.

  19. Evaluation of Pseudomonas aeruginosa an innovative bioremediation tool in multi metals ions from simulated system using multi response methodology.

    Science.gov (United States)

    Singh, Rajesh; Bishnoi, Narsi R; Kirrolia, Anita

    2013-06-01

    Under certain conditions bacteria can act as a good biosorbent towards heavy metals in simultaneous removal from effluents. The present study explores overlay plots of multi response surface methodology for simulated wastewater treatment potential. Pseudomonas aeruginosa was used for bioremediation of metallic ions, where removal of Cd (80-90%), Mn (85-90%), Fe (50-55%), Cr (70-75%) can be achieved by fixing the pH, oxidation reduction potential (mV) and one of the metallic constituent in the simulated effluent. The metal ions Cd and Cr (T), Fe and ORP (mV) are relatively closely located to each other in the loading plot indicating co-variance between these components. However Cr(VI) transformation and Mn removal are distantly placed in the bi-plot indicating the existed significant difference. Elevated reductase enzyme activity (31.75 μg/minmg) observed in the isolate showing the ability to effectively reduce metals ions.

  20. Heavy metal removal from sediments by biosurfactants.

    Science.gov (United States)

    Mulligan, C N; Yong, R N; Gibbs, B F

    2001-07-30

    Batch washing experiments were used to evaluate the feasibility of using biosurfactants for the removal of heavy metals from sediments. Surfactin from Bacillus subtilis, rhamnolipids from Pseudomonas aeruginosa and sophorolipid from Torulopsis bombicola were evaluated using a metal-contaminated sediment (110mg/kg copper and 3300mg/kg zinc). A single washing with 0.5% rhamnolipid removed 65% of the copper and 18% of the zinc, whereas 4% sophorolipid removed 25% of the copper and 60% of the zinc. Surfactin was less effective, removing 15% of the copper and 6% of the zinc. The technique of ultrafiltration and zeta potential measurements were used to determine the mechanism of metal removal by the surfactants. It was then postulated that metal removal by the biosurfactants occurs through sorption of the surfactant on to the soil surface and complexation with the metal, detachment of the metal from the soil into the soil solution and hence association with surfactant micelles. Sequential extraction procedures were used on the sediment to determine the speciation of the heavy metals before and after surfactant washing. The carbonate and oxide fractions accounted for over 90% of the zinc present in the sediments. The organic fraction constituted over 70% of the copper. Sequential extraction of the sediments after washing with the various surfactants indicated that the biosurfactants, rhamnolipid and surfactin could remove the organically-bound copper and that the sophorolipid could remove the carbonate and oxide-bound zinc. Therefore, heavy metal removal from sediments is feasible and further research will be conducted.

  1. Antibacterial activity of Lawsonia inermis Linn (Henna) against Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    Habbal O; Hasson SS; El-Hag AH; Al-Mahrooqi Z; Al-Hashmi N; Al-Bimani Z; MS Al-Balushi; Al-Jabri AA

    2011-01-01

    Objective: To investigate the antibacterial activity of henna (Lawsonia inermis Linn) obtained from different regions of Oman against a wide array of micro-organisms. Methods: Fresh henna samples were obtained from different regions of Oman as leaves and seeds. 100 g fresh and dry leaves and 50 g of fresh and dry seeds were separately soaked in 500 mL of ethanol for three days, respectively, with frequent agitation. The mixture was filtered, and the crude extract was collected. The crude extract was then heated, at 48 ℃ in a water bath to evaporate its liquid content. The dry crude henna extract was then tested for its antibacterial activity using well-diffusion antibiotic susceptibility technique. Henna extracts were investigated for their antibacterial activity at different concentrations against a wide array of different micro-organisms including a laboratory standard bacterial strain of Pseudomonas aeruginosa (NCTC 10662) (P. aeruginosa) and eleven fresh clinical isolates of P. aeruginosa obtained from patients attending the Sultan Qaboos University Hospital (SQUH). 2-Hydroxy-p-Nathoqinone-Tech (2-HPNT, MW=174.16, C10H6O3) was included as control (at 50% concentration) along with the henna samples tested. Results: Henna samples demonstrated antibacterial activity against all isolates but the highest susceptibility was against P. aeruginosa with henna samples obtained from Al-sharqyia region. Conclusions: Omani henna from Al-sharqyia region demonstrates high in vitro anti-P. aeruginosa activity compared with many henna samples from different regions of Oman.

  2. The Genomic Basis of Evolutionary Innovation in Pseudomonas aeruginosa.

    Science.gov (United States)

    Toll-Riera, Macarena; San Millan, Alvaro; Wagner, Andreas; MacLean, R Craig

    2016-05-01

    Novel traits play a key role in evolution, but their origins remain poorly understood. Here we address this problem by using experimental evolution to study bacterial innovation in real time. We allowed 380 populations of Pseudomonas aeruginosa to adapt to 95 different carbon sources that challenged bacteria with either evolving novel metabolic traits or optimizing existing traits. Whole genome sequencing of more than 80 clones revealed profound differences in the genetic basis of innovation and optimization. Innovation was associated with the rapid acquisition of mutations in genes involved in transcription and metabolism. Mutations in pre-existing duplicate genes in the P. aeruginosa genome were common during innovation, but not optimization. These duplicate genes may have been acquired by P. aeruginosa due to either spontaneous gene amplification or horizontal gene transfer. High throughput phenotype assays revealed that novelty was associated with increased pleiotropic costs that are likely to constrain innovation. However, mutations in duplicate genes with close homologs in the P. aeruginosa genome were associated with low pleiotropic costs compared to mutations in duplicate genes with distant homologs in the P. aeruginosa genome, suggesting that functional redundancy between duplicates facilitates innovation by buffering pleiotropic costs.

  3. Influence of Pseudomonas aeruginosa on exacerbation in patients with bronchiectasis

    Directory of Open Access Journals (Sweden)

    Kiran Chawla

    2015-01-01

    Full Text Available Background: A majority of the studies done on the western population have shown that Pseudomonas aeruginosa causes many severe infections in patients with bronchiectasis as compared to other pathogens. There is scarcity of similar data from the Asian population. Materials and Methods: A prospective study was undertaken to identify the various pathogens isolated from the respiratory samples of 117 patients with bronchiectasis from south India and to compare the clinicomicrobiological profile of infections caused by P. aeruginosa and other respiratory pathogens. Results: The respiratory pathogens were isolated from 63 (53.8% patients. P. aeruginosa was the most common isolate (46.0% followed by Klebsiella pneumoniae (14.3% and other pathogenic bacteria. Patients included in the P. aeruginosa group had a higher number of exacerbations (p: 0.008, greater number of hospital admissions (p: 0.007, a prolonged hospital stay (p: 0.03, and poor lung function, compared to the patients infected with the non-Pseudomonas group. Conclusion: It is necessary to investigate the etiology of respiratory tract infections among bronchiectasis patients followed by the prompt management of cases diagnosed with P. aeruginosa infections, so as to lower the morbidity and have a better prognosis.

  4. Balneotherapy is a potential risk factor for Pseudomonas aeruginosa colonization

    Directory of Open Access Journals (Sweden)

    Gabriela Deutsch

    Full Text Available ABSTRACT The practice of immersion in burn patient has been abandoned in many parts of the world but in Brazil it is still common. The aim of this study was to ascertain if balneotherapy is a risk factor for Pseudomonas aeruginosa colonization in thermally injured patients. Eighteen patients from a Burn Center were studied for 14 weeks for Pseudomonas aeruginosa. Samples were collected by swabbing the exudate of wounds, before and after giving bath to the patients and from balneotherapy table. Pulsed-field gel electrophoresis was used to determine bacterial genetic relatedness. Thirty-seven P. aeruginosa isolates were detected from 292 swabs collected from patients' burn surface area and from the balneotherapy table. Profile analysis of P. aeruginosa DNA fragmentation showed 10 clones among the 37 strains analyzed. Type A is the most prevalent clone, with 23 strains distributed into eight subtypes. These were present in the swabs collected, before and after the patients' bath, from the surface of the bath table, suggesting that there was cross-contamination between the patients in different ways. This work demonstrates that balneotherapy is a risk factor in the Burn Center studied, because the same clone was found among P. aeruginosa isolates collected at various points and times.

  5. Genetic and functional diversity of Pseudomonas aeruginosa lipopolysaccharide

    Directory of Open Access Journals (Sweden)

    Joseph S. Lam

    2011-06-01

    Full Text Available Lipopolysccharide (LPS is an integral component of the Pseudomonas aeruginosa cell envelope, occupying the outer leaflet of the outer membrane in this Gram-negative opportunistic pathogen. It is important for bacteria-host interactions and has been shown to be a major virulence factor for this organism. Structurally, P. aeruginosa LPS is composed of three domains, namely, lipid A, core oligosaccharide, and the distal O antigen (O-Ag. Most P. aeruginosa strains produce two distinct forms of O-Ag, one a homopolymer of D-rhamnose that is a common polysaccharide antigen (CPA, formerly termed A band, and the other a heteropolymer of three to five distinct (and often unique dideoxy sugars in its repeat units, known as O-specific antigen (OSA, formerly termed B band. Compositional differences in the O units among the OSA from different strains form the basis of the International Antigenic Typing Scheme for classification via serotyping of different strains of P. aeruginosa. The focus of this review is to provide state-of-the-art knowledge on the genetic and resultant functional diversity of LPS produced by P. aeruginosa. The underlying factors contributing to this diversity will be thoroughly discussed and presented in the context of its contributions to host-pathogen interactions and the control/prevention of infection.

  6. A Network Biology Approach to Denitrification in Pseudomonas aeruginosa

    Science.gov (United States)

    Arat, Seda; Bullerjahn, George S.; Laubenbacher, Reinhard

    2015-01-01

    Pseudomonas aeruginosa is a metabolically flexible member of the Gammaproteobacteria. Under anaerobic conditions and the presence of nitrate, P. aeruginosa can perform (complete) denitrification, a respiratory process of dissimilatory nitrate reduction to nitrogen gas via nitrite (NO2), nitric oxide (NO) and nitrous oxide (N2O). This study focuses on understanding the influence of environmental conditions on bacterial denitrification performance, using a mathematical model of a metabolic network in P. aeruginosa. To our knowledge, this is the first mathematical model of denitrification for this bacterium. Analysis of the long-term behavior of the network under changing concentration levels of oxygen (O2), nitrate (NO3), and phosphate (PO4) suggests that PO4 concentration strongly affects denitrification performance. The model provides three predictions on denitrification activity of P. aeruginosa under various environmental conditions, and these predictions are either experimentally validated or supported by pertinent biological literature. One motivation for this study is to capture the effect of PO4 on a denitrification metabolic network of P. aeruginosa in order to shed light on mechanisms for greenhouse gas N2O accumulation during seasonal oxygen depletion in aquatic environments such as Lake Erie (Laurentian Great Lakes, USA). Simulating the microbial production of greenhouse gases in anaerobic aquatic systems such as Lake Erie allows a deeper understanding of the contributing environmental effects that will inform studies on, and remediation strategies for, other hypoxic sites worldwide. PMID:25706405

  7. Review: Antibiotic discovery in the age of structural biology - a comprehensive overview with special reference to development of drugs for the treatment of Pseudomonas aeruginosa infection.

    Science.gov (United States)

    Koehnke, Alessa; Friedrich, Reinhard E

    2015-01-01

    Due to the persistence and spread of antibiotic resistance, the discovery and exploitation of new antibiotic targets should be the subject of intensive research. Effective strategies are required to develop antibiotic alternatives. Antibiotics that act on new targets or via novel mechanisms have the greatest likelihood of overcoming resistance. In particular, there is a lack of specific antibiotics for Pseudomonas aeruginosa, one of the leading causes of healthcare-associated infections, exhibiting high resistance levels. Herein we describe how structure-based drug design can be used to achieve new antibiotics for the treatment of Pseudomonas aeruginosa infection, using an essential enzyme of the fatty acid synthesis pathway from P. aeruginosa as an example.

  8. Molecular detection of an atypical, highly resistant, clonal Pseudomonas aeruginosa isolate in cystic fibrosis patients.

    LENUS (Irish Health Repository)

    Keating, Deirdre

    2013-03-01

    The identification of Pseudomonas aeruginosa (P. aeruginosa) isolates in sputum from cystic fibrosis (CF) patients can be challenging due to the multitude of phenotypic changes isolates undergo during adaptation to the microenvironment of the CF lung.

  9. Initial Pseudomonas aeruginosa infection in patients with cystic fibrosis: characteristics of eradicated and persistent isolates

    DEFF Research Database (Denmark)

    Tramper-Stranders, G. A.; van der Ent, C. K.; Molin, Søren;

    2012-01-01

    Clin Microbiol Infect 2012; 18: 567574 Abstract Despite intensive eradication therapy, some CF patients with early Pseudomonas aeruginosa infection rapidly develop a chronic infection. To elucidate factors associated with this persistence, bacterial characteristics of early P. aeruginosa isolates...

  10. Diversity of metabolic profiles of cystic fibrosis Pseudomonas aeruginosa during the early stages of lung infection

    DEFF Research Database (Denmark)

    Jørgensen, Karin Meinike; Wassermann, Tina; Johansen, Helle Krogh;

    2015-01-01

    Pseudomonas aeruginosa is the dominant pathogen infecting the airways of cystic fibrosis (CF) patients. During the intermittent colonization phase, P. aeruginosa resembles environmental strains but later evolves to the chronic adapted phenotype characterized by resistance to antibiotics...

  11. Ambroxol inhibits mucoid conversion of Pseudomonas aeruginosa and contributes to the bactericidal activity of ciprofloxacin against mucoid P. aeruginosa biofilms.

    Science.gov (United States)

    Wang, Wenlei; Yu, Jialin; He, Yu; Wang, Zhengli; Li, Fang

    2016-07-01

    Pseudomonas aeruginosa is an opportunistic human pathogen that can cause severe infections in immunocompromised individuals. Because it forms biofilms, which protect against host immune attack and increase resistance to conventional antibiotics, mucoid P. aeruginosa is nearly impossible to eradicate. Moreover, mucoid conversion of P. aeruginosa in cystic fibrosis (CF) patients leads to poor outcomes. This conversion is mainly due to mucA gene mutation, which is thought to be induced by polymorphonuclear leukocytes (PMNs) and the reactive oxygen species they release. Ambroxol, a mucolytic agent with antioxidant characteristics, is used clinically, and this compound has recently been demonstrated to possess anti-biofilm properties. In this study, we found that ambroxol inhibits the H2 O2 -mediated conversion of P. aeruginosa from a non-mucoid to a mucoid phenotype, an effect that is due to its antioxidant property against H2 O2 . Furthermore, the bactericidal activity of ciprofloxacin against mucoid P. aeruginosa biofilms was increased in vitro when used in combination with ambroxol.

  12. Effects of antibiotics on quorum sensing in pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Skindersø, Mette Elena; Alhede, Morten; Phipps, Richard Kerry

    2008-01-01

    . Three of the antibiotics tested, AZM, ceftazidime (CFT), and ciprofloxacin (CPR), were very active in the assay and were further examined for their effects on QS-regulated virulence factor production in P. aeruginosa. The effects of the three antibiotics administered at subinhibitory concentrations were...... in animal infection models. Treatment of cystic fibrosis (CF) patients chronically infected with P. aeruginosa with the macrolide antibiotic azithromycin (AZM) has been demonstrated to improve the clinical outcome. Several studies indicate that AZM may accomplish its beneficial action in CF patients...... by impeding QS, thereby reducing the pathogenicity of P. aeruginosa. This led us to investigate whether QS inhibition is a common feature of antibiotics. We present the results of a screening of 12 antibiotics for their QS-inhibitory activities using a previously described QS inhibitor selector 1 strain...

  13. Vaccines for preventing infection with Pseudomonas aeruginosa in cystic fibrosis

    DEFF Research Database (Denmark)

    Johansen, Helle Krogh; Gøtzsche, Peter C

    2015-01-01

    BACKGROUND: Chronic pulmonary infection in cystic fibrosis results in progressive lung damage. Once colonisation of the lungs with Pseudomonas aeruginosa occurs, it is almost impossible to eradicate. Vaccines, aimed at reducing infection with Pseudomonas aeruginosa, have been developed...... in the vaccine group and 91 (1 severe) in the control group. In this large trial of a vaccine developed against flagella antigens, antibody titres against the epitopes contained in the vaccine were higher in the vaccine group compared to the placebo group (P Vaccines against....... This is an update of a previously published review. OBJECTIVES: To assess the effectiveness of vaccination against Pseudomonas aeruginosa in cystic fibrosis. SEARCH METHODS: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register using the terms vaccines AND pseudomonas (last search 30...

  14. Subtilase SprP exerts pleiotropic effects in Pseudomonas aeruginosa.

    Science.gov (United States)

    Pelzer, Alexander; Polen, Tino; Funken, Horst; Rosenau, Frank; Wilhelm, Susanne; Bott, Michael; Jaeger, Karl-Erich

    2014-02-01

    The open reading frame PA1242 in the genome of Pseudomonas aeruginosa PAO1 encodes a putative protease belonging to the peptidase S8 family of subtilases. The respective enzyme termed SprP consists of an N-terminal signal peptide and a so-called S8 domain linked by a domain of unknown function (DUF). Presumably, this DUF domain defines a discrete class of Pseudomonas proteins as homologous domains can be identified almost exclusively in proteins of the genus Pseudomonas. The sprP gene was expressed in Escherichia coli and proteolytic activity was demonstrated. A P. aeruginosa ∆sprP mutant was constructed and its gene expression pattern compared to the wild-type strain by genome microarray analysis revealing altered expression levels of 218 genes. Apparently, SprP is involved in regulation of a variety of different cellular processes in P. aeruginosa including pyoverdine synthesis, denitrification, the formation of cell aggregates, and of biofilms.

  15. Novel Targets for Treatment of Pseudomonas aeruginosa Biofilms

    DEFF Research Database (Denmark)

    Alhede, Morten; Alhede, Maria; Bjarnsholt, Thomas

    2014-01-01

    Pseudomonas aeruginosa causes infection in all parts of the human body. The bacterium is naturally resistant to a wide range of antibiotics. In addition to resistance mechanisms such as efflux pumps, the ability to form aggregates, known as biofilm, further reduces Pseudomonas aeruginosa’s suscep......Pseudomonas aeruginosa causes infection in all parts of the human body. The bacterium is naturally resistant to a wide range of antibiotics. In addition to resistance mechanisms such as efflux pumps, the ability to form aggregates, known as biofilm, further reduces Pseudomonas aeruginosa......’s susceptibility to antibiotics. The presence of such biofilms is acknowledged to equal a persistent infection due to their inherent high tolerance to all antimicrobials and immune cells. In this chapter we discuss the mechanisms of biofilm tolerance. The latest biofilm research is reviewed and future treatment...

  16. Hemorrhagic pneumonia in mink caused by Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Salomonsen, Charlotte Mark

    Hemorrhagic pneumonia in mink is an acute and fatal disease caused by Pseudomonas aeruginosa. The mink are typically found dead without prior clinical symptoms. The disease can be highly contagious and varying mortalities on the farm level has been reported. Hemorrhagic pneumonia in mink...... in hemorrhagic pneumonia caused by P. aeruginosa and E. coli in diagnostic material. The distribution of the two pathogens is visualized using fluorescence in situ hybridization (FISH). Two histological patterns were observed in the work presented in Article II; one was very hemorrhagic with few bacteria while...... is seasonal with outbreaks almost exclusively occurring from September to November in Denmark. In human medicine, P. aeruginosa is regarded as a pathogen for immune compromised individuals but no underlying disease or immune defect has been identified in mink dying of hemorrhagic pneumonia. In fact, little...

  17. Effects of ambroxol on alginate of mature Pseudomonas aeruginosa biofilms.

    Science.gov (United States)

    Li, Fang; Yu, Jialin; Yang, Hua; Wan, Zhenyan; Bai, Dan

    2008-07-01

    Biofilm-forming bacteria Pseudomonas aeruginosa is a common pathogen in mechanically ventilated newborns, which can cause life-threatening infections. Alginate of mucoid Pseudomonas aeruginosa biofilms is considered an important virulence factor which contributes to the resistance to antibiotics. Traditionally, ambroxol is widely used in newborns with lung problems as a mucolytic agent and antioxidant agent as well. And there are few studies that demonstrated the anti-biofilm activity of ambroxol. In this study, we found that ambroxol can affect the structure of mucoid Pseudomonas aeruginosa biofilms. Further, we found that ambroxol reduces the production of alginate, the expression of the important genes and the activity of key enzyme guanosine diphospho-D-mannose dehydrogenase (GDP-mannose dehydrogenase; GMD) which were involved in alginate biosynthesis.

  18. Outbreak of Pseudomonas aeruginosa bacteraemia in a haematology department

    DEFF Research Database (Denmark)

    Rasmussen, Benjamin Schnack; Christensen, Nikolas; Sørensen, Jan

    2015-01-01

    that tested positive for P. aeruginosa were collected from the laboratory information system (MADS, Skejby Hospital, Aarhus, Denmark). Environmental samples were obtained from shower heads in the department. The genotype was established by pulse field gel electrophoresis (PFGE). An audit was conducted during......INTRODUCTION: Infection by Pseudomonas aeruginosa represents a major cause of morbidity and mortality among immunocompromised patients. In Denmark, an increase in P. aeruginosa isolates from blood cultures from a haematology department prompted a hygienic audit in 2007. METHODS: Blood cultures...... the outbreak and 12 months later. The audits were conducted by the method of direct observation. RESULTS: Several PFGE types were involved with no clear association to isolates from environmental samples. The audit revealed poor hygiene related to the handling of central venous catheters. After optimising...

  19. Role of quorum sensing by Pseudomonas aeruginosa in microbial keratitis and cystic fibrosis

    DEFF Research Database (Denmark)

    Willcox, M.D.P.; Zhu, H.; Conibear, T.C.R.

    2008-01-01

    Pseudomonas aeruginosa is a ubiquitous bacterium that causes opportunistic infections in a range of host tissues and organs. Infections by P. aeruginosa are difficult to treat and hence there is interest in the development of effective therapeutics. One of the key mechanisms that P. aeruginosa us...

  20. The evolution and adaptation of clinical Pseudomonas aeruginosa isolates from early cystic fibrosis infections

    DEFF Research Database (Denmark)

    Lindegaard, Mikkel

    Pseudomonas aeruginosa is a major cause of morbidity and mortality in cystic fibrosis (CF) patients. P. aeruginosa infects the CF airways and establishes chronic infections that can last for a lifetime during which P.aeruginosa evolves in order to adapt to the environment.In this PhD thesis, we...

  1. A case of failed eradication of cystic fibrosis-related sinus colonisation by Pseudomonas aeruginosa.

    LENUS (Irish Health Repository)

    Linnane, Barry

    2015-10-01

    Pseudomonas aeruginosa is a pathogen associated with cystic fibrosis that has potential to decrease lung function and cause respiratory failure. Paranasal sinuses are increasingly recognised as potential reservoirs for intermittent colonisation by P. aeruginosa. This case documents investigation and outcome of P. aeruginosa recurrence in a male paediatric patient over an eight year period.

  2. Force microscopic and thermodynamic analysis of the adhesion between Pseudomonas aeruginosa and Candida albicans

    NARCIS (Netherlands)

    Ovchinnikova, Ekaterina S.; Krom, Bastiaan P.; van der Mei, Henny C.; Busscher, Henk J.

    2012-01-01

    Pseudomonas aeruginosa expresses a plethora of virulence factors and many species have developed warning systems to detect and evade P. aeruginosa. Candida albicans detects P. aeruginosa by sensing the secreted bacterial quorum sensing molecule 3OC(12)HSL and responds by reverting to the yeast morph

  3. Simultaneous Removal of Phenol and Dissolved Solids from Wastewater Using Multichambered Microbial Desalination Cell.

    Science.gov (United States)

    Pradhan, Harapriya; Jain, Sumat Chand; Ghangrekar, Makarand M

    2015-12-01

    Microbial desalination cell (MDC) has great potential toward direct electricity generation from wastewater and concurrent desalination through potential difference developed due to microbial activity. Degradation of phenol by isolate Pseudomonas aeruginosa in anodic chamber and simultaneous desalination of water in middle desalination chamber of multichamber MDC is demonstrated in this study. Performance of the MDCs with different anodic inoculum conditions, namely pure culture of P. aeruginosa (MDC-1), 50 % v/v mixture of P. aeruginosa and anaerobic mixed consortia (MDC-2) and anaerobic mixed consortia (MDC-3), was evaluated to compare the phenol degradation in anodic chamber, bioelectricity generation, and simultaneous total dissolved solids (TDS) removal from saline water in desalination chamber. Synergistic effect between P. aeruginosa and mixed anaerobic consortia as inoculum was evident in MDC-2 demonstrating phenol degradation of 90 %, TDS removal of 75 % in 72 h of reaction time along with higher power generation of 27.5 mW/m(2) as compared to MDC-1 (95 %, 64 %, 12.8 mW/m(2), respectively) and MDC-3 (58 %, 52 %, 4.8 mW/m(2), respectively). The results illustrate that the multichamber MDC-2 is effective for simultaneous removal of phenol and dissolved solids contained in industrial wastewaters.

  4. [Antibiotic activity of P. aeruginosa against MRSA and Candida albicans].

    Science.gov (United States)

    Kondo, Shigemi; Sato, Naotake; Yamada, Toshihiko; Miyazaki, Sakiko; Oguri, Toyoko; Igari, Jun

    2002-04-01

    The antibiotic activity demonstrated by P. aeruginosa (Bacillus pyocyaneus) has been reported more than one hundred years ago by Emmerich et al (1899). Studies on such bacterial interference between P. aeruginosa and other pathogenic bacteria or fungi have not been extensively reported in recent years. In this paper, we report on the anti MRSA activity and anti Candida activity demonstrated by clinical isolates of P. aeruginosa (34 strains). The antibiotic activity was tested by reversed agar plate method, as previously reported, and the degree of the activity was expressed as the diameter of the zone of growth inhibition. The stability of both anti MRSA activity and anti Candida activity was evaluated at the time after 24 and 48-hr incubation. Also the effect of agar plate with or without 5% sheep blood on antibiotic activity was evaluated. Strong anti MRSA activity and anti Candida activity was shown at the time after 24-hr incubation. At the time after 48-hr incubation, anti MRSA activities were significantly suppressed but anti Candida activities were persisted. The inhibitory activity was correlated with dye production of P. aeruginosa. Some strains having non or weak dye production, showed the inhibitory activity by 48-hr incubation. Result from these strains without suppression of anti Candida activity by additional blood may suggest that the existence of a new factor produced by P. aeruginosa. Because of frequent isolation of MRSA or Candida from clinical materials, we must consider bacterial flora and bacterial interference against pathogenic bacteria at the time of the antibiotic choice for the patients infected or colonized with P. aeruginosa.

  5. Prevalence and analysis of Pseudomonas aeruginosa in chinchillas

    Directory of Open Access Journals (Sweden)

    Aoyama Naoki

    2010-11-01

    Full Text Available Abstract Background Chinchillas (Chinchilla laniger are popular as pets and are often used as laboratory animals for various studies. Pseudomonas aeruginosa is a major infectious agent that causes otitis media, pneumonia, septicaemia enteritis, and sudden death in chinchillas. This bacterium is also a leading cause of nosocomial infections in humans. To prevent propagation of P. aeruginosa infection among humans and animals, detailed characteristics of the isolates, including antibiotic susceptibility and genetic features, are needed. In this study, we surveyed P. aeruginosa distribution in chinchillas bred as pets or laboratory animals. We also characterized the isolates from these chinchillas by testing for antibiotic susceptibility and by gene analysis. Results P. aeruginosa was isolated from 41.8% of the 67 chinchillas included in the study. Slide agglutination and pulsed-field gel electrophoresis discriminated 5 serotypes and 7 unique patterns, respectively. For the antibiotic susceptibility test, 40.9% of isolates were susceptible to gentamicin, 77.3% to ciprofloxacin, 77.3% to imipenem, and 72.7% to ceftazidime. DNA analyses confirmed that none of the isolates contained the gene encoding extended-spectrum β-lactamases; however, 2 of the total 23 isolates were found to have a gene similar to the pilL gene that has been identified in the pathogenicity island of a clinical isolate of P. aeruginosa. Conclusions P. aeruginosa is widely spread in chinchillas, including strains with reduced susceptibility to the antibiotics and highly virulent strains. The periodic monitoring should be performed to help prevent the propagation of this pathogen and reduce the risk of infection from chinchillas to humans.

  6. Bioleaching of copper oxide ore by P seudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    MA Shabani; M Irannajad; AR Azadmehr; M Meshkini

    2013-01-01

    Bioleaching is an environmentally friendly method for extraction of metal from ores. In this study, bioleaching of copper oxide ore by Pseudomonas aeruginosa was investigated. Pseudomonas aeruginosa is a heterotrophic bacterium that can produce various organic acids in an appropriate culture medium, and these acids can operate as leaching agents. The parameters, such as particle size, glucose percentage in the culture medium, bioleaching time, and solid/liquid ratio were optimized. Optimum bioleaching conditions were found as follows: particle size of 150-177 μm, glucose percentage of 6%, bioleaching time of 8 d, and solid/liquid ratio of 1:80. Under these conditions, 53%of copper was extracted.

  7. Anionic fluoroquinolones as antibacterials against biofilm-producing Pseudomonas aeruginosa.

    Science.gov (United States)

    Long, Timothy E; Keding, Lexie C; Lewis, Demetria D; Anstead, Michael I; Withers, T Ryan; Yu, Hongwei D

    2016-02-15

    Pseudomonas aeruginosa is a common biofilm-forming bacterial pathogen implicated in diseases of the lungs. The extracellular polymeric substances (EPS) of respiratory Pseudomonas biofilms are largely comprised of anionic molecules such as rhamnolipids and alginate that promote a mucoid phenotype. In this Letter, we examine the ability of negatively-charged fluoroquinolones to transverse the EPS and inhibit the growth of mucoid P. aeruginosa. Anionic fluoroquinolones were further compared with standard antibiotics via a novel microdiffusion assay to evaluate drug penetration through pseudomonal alginate and respiratory mucus from a patient with cystic fibrosis.

  8. Characterization of temporal protein production in Pseudomonas aeruginosa biofilms.

    Science.gov (United States)

    Southey-Pillig, Christopher J; Davies, David G; Sauer, Karin

    2005-12-01

    Phenotypic and genetic evidence supporting the notion of biofilm formation as a developmental process is growing. In the present work, we provide additional support for this hypothesis by identifying the onset of accumulation of biofilm-stage specific proteins during Pseudomonas aeruginosa biofilm maturation and by tracking the abundance of these proteins in planktonic and three biofilm developmental stages. The onset of protein production was found to correlate with the progression of biofilms in developmental stages. Protein identification revealed that proteins with similar function grouped within similar protein abundance patterns. Metabolic and housekeeping proteins were found to group within a pattern separate from virulence, antibiotic resistance, and quorum-sensing-related proteins. The latter were produced in a progressive manner, indicating that attendant features that are characteristic of biofilms such as antibiotic resistance and virulence may be part of the biofilm developmental process. Mutations in genes for selected proteins from several protein production patterns were made, and the impact of these mutations on biofilm development was evaluated. The proteins cytochrome c oxidase, a probable chemotaxis transducer, a two-component response regulator, and MexH were produced only in mature and late-stage biofilms. Mutations in the genes encoding these proteins did not confer defects in growth, initial attachment, early biofilm formation, or twitching motility but were observed to arrest biofilm development at the stage of cell cluster formation we call the maturation-1 stage. The results indicated that expression of theses genes was required for the progression of biofilms into three-dimensional structures on abiotic surfaces and the completion of the biofilm developmental cycle. Reverse transcription-PCR analysis confirmed the detectable change in expression of the respective genes ccoO, PA4101, and PA4208. We propose a possible mechanism for the

  9. UV-induced endonuclease III-sensitive sites at the mating type loci in Saccharomyces cerevisiae are repaired by nucleotide excision repair: RAD7 and RAD16 are not required for their removal from HML alpha.

    Science.gov (United States)

    Reed, S H; Boiteux, S; Waters, R

    1996-03-01

    Ultraviolet irradiation of DNA induces cyclobutane pyrimidine dimers (CPDs) 6-4'-(pyrimidine 2'-one) pyrimidines and pyrimidine hydrates. The dimer is the major photoproduct, and is specifically recognized by endonuclease V of phage T4. Pyrimidine hydrates represent a small fraction of the total photoproducts, and are substrates for endonuclease III of Escherichia coli. We used these enzymes to follow the fate of their substrates in the mating type loci of Saccharomyces cerevisiae. In a RAD strain, CPSs in the transcriptionally active MAT alpha locus are preferentially repaired relative to the inactive HML alpha locus, whilst repair of endonuclease III-sensitive sites is not preferential. The rad1, 2, 3 and 4 mutants, which lack factors that are essential for the incision step of nucleotide excision repair (NER), repair neither CPDs nor endonuclease III-sensitive sites, clearly showing that these lesions are repaired by by NER pathway. Previously it had been shown that the products of the RAD7 and RAD16 genes are required for the NER of CPDs from the HML alpha locus. We show that, in the same locus, these gene products are not needed for removal of endonuclease III-sensitive sites by the same mechanism. This indicates that the components required for NER differ depending on either the type of lesion encountered or on the specific location of the lesion within the genome.

  10. Eradication of Pseudomonas aeruginosa cells by cathodic electrochemical currents delivered with graphite electrodes.

    Science.gov (United States)

    Niepa, Tagbo H R; Wang, Hao; Gilbert, Jeremy L; Ren, Dacheng

    2017-03-01

    Antibiotic resistance is a major challenge to the treatment of bacterial infections associated with medical devices and biomaterials. One important intrinsic mechanism of such resistance is the formation of persister cells that are phenotypic variants of microorganisms and highly tolerant to antibiotics. Recently, we reported a new approach to eradicating persister cells of Pseudomonas aeruginosa using low-level direct electrochemical current (DC) and synergy with the antibiotic tobramycin. To further understand the underlying mechanism and develop this technology toward possible medical applications, we investigated the electricidal activities of non-metallic biomaterial on persister and biofilm cells of P. aeruginosa using graphite-based TGON™ 805 electrodes. We employed both single and dual chamber systems to compare electrochemical factors of TGON and stainless steel 304 electrodes. The results revealed that TGON-based treatments were highly effective against P. aeruginosa persister cells. In the single chamber system, complete eradication of planktonic persister cells (corresponding to a 7-log killing) was achieved with 70μA/cm(2) DC using TGON electrodes within 40min of treatment, while the cell viability in biofilms was reduced by 2 logs within 1h. The killing effects were dose and time dependent with higher current densities requiring less time. Moreover, reduction reactions were found more effective than oxidation reactions, confirming that metal cations are not indispensable, although they may facilitate cell killing. The findings of this study can help develop electrochemical technologies to eradicate persister and biofilm cells for more effective treatment of medical device and biomaterial associated infections.

  11. Gene expression in Pseudomonas aeruginosa swarming motility

    Directory of Open Access Journals (Sweden)

    Déziel Eric

    2010-10-01

    Full Text Available Abstract Background The bacterium Pseudomonas aeruginosa is capable of three types of motilities: swimming, twitching and swarming. The latter is characterized by a fast and coordinated group movement over a semi-solid surface resulting from intercellular interactions and morphological differentiation. A striking feature of swarming motility is the complex fractal-like patterns displayed by migrating bacteria while they move away from their inoculation point. This type of group behaviour is still poorly understood and its characterization provides important information on bacterial structured communities such as biofilms. Using GeneChip® Affymetrix microarrays, we obtained the transcriptomic profiles of both bacterial populations located at the tip of migrating tendrils and swarm center of swarming colonies and compared these profiles to that of a bacterial control population grown on the same media but solidified to not allow swarming motility. Results Microarray raw data were corrected for background noise with the RMA algorithm and quantile normalized. Differentially expressed genes between the three conditions were selected using a threshold of 1.5 log2-fold, which gave a total of 378 selected genes (6.3% of the predicted open reading frames of strain PA14. Major shifts in gene expression patterns are observed in each growth conditions, highlighting the presence of distinct bacterial subpopulations within a swarming colony (tendril tips vs. swarm center. Unexpectedly, microarrays expression data reveal that a minority of genes are up-regulated in tendril tip populations. Among them, we found energy metabolism, ribosomal protein and transport of small molecules related genes. On the other hand, many well-known virulence factors genes were globally repressed in tendril tip cells. Swarm center cells are distinct and appear to be under oxidative and copper stress responses. Conclusions Results reported in this study show that, as opposed to

  12. Pseudomonas aeruginosa en agua y leche cruda: informe preliminar Pseudomonas aeruginosa in water and raw milk: preliminary report

    Directory of Open Access Journals (Sweden)

    M. S Iramain

    2005-12-01

    Full Text Available El objetivo del trabajo fue determinar la presencia de Pseudomonas aeruginosa en el agua utilizada en las tareas relacionadas al ordeño y en leche de tanque, para establecer una posible vinculación entre la contaminación del agua y de la leche cruda con esta bacteria, en tambos de la provincia de Buenos Aires. Se muestrearon y analizaron 122 tambos, obteniéndose muestras de 111 perforaciones, 92 tanques de almacenamiento de agua y 122 de leche de tanque según normas de referencia. En todos los casos se determinó la presencia de P. aeruginosa, hallándose en el 27% de las muestras de perforaciones y en el 34% de los tanques de almacenamiento. Solamente 4 establecimientos presentaron P.aeruginosa en leche de tanque, pudiéndose constatar que en tres de ellos se realizaban prácticas operativas que ponían en contacto la leche con el agua contaminada. Una vez eliminadas éstas prácticas no fue posible hallar P. aeruginosa en la leche de los tanques.The aim of this study was to determine the presence of Pseudomonas aeruginosa in the water used in milking practices and in bulk tank milk, to establish a possible relationship between water and raw bulk tank milk contamination with this bacteria, in dairy farms of Buenos Aires province. Samples from 122 dairy farms were analyzed for P. aeruginosa according to reference methods, getting 111 underground water samples and 92 water storage tank samples and 122 bulk tank milk samples. Twenty seven per cent of underground water samples were positive for P. aeruginosa as well as 34 % of storage tank samples. The bacteria was present in only 4 dairy farms bulk tanks. It was determined that in 3 of them milking management practices allowed the milk to get in contact with contaminated water. Once these practices were eliminated, no P. aeruginosa was found in bulk tank milk samples

  13. Detection of boron removal capacities of different microorganisms in wastewater and effective removal process.

    Science.gov (United States)

    Laçin, Bengü; Ertit Taştan, Burcu; Dönmez, Gönül

    2015-01-01

    In this study boron removal capacities of different microorganisms were tested. Candida tropicalis, Rhodotorula mucilaginosa, Micrococcus luteus, Bacillus thuringiensis, Bacillus cereus, Bacillus megaterium, Bacillus pumilus, Pseudomonas aeruginosa and Aspergillus versicolor were examined for their boron bioaccumulation capacities in simulated municipal wastewater. A. versicolor and B. cereus were found as the most boron-tolerant microorganisms in the experiments. Also boron bioaccumulation yield of A. versicolor was 49.25% at 15 mg/L boron concentration. On the other hand biosorption experiments revealed that A. versicolor was more capable of boron removal in inactive form at the highest boron concentrations. In this paper maximum boron bioaccumulation yield was detected as 39.08% at 24.17 mg/L and the maximum boron biosorption yield was detected as 41.36% at 24.01 mg/L boron concentrations.

  14. Secretory IgA as a diagnostic tool for Pseudomonas aeruginosa respiratory colonization

    DEFF Research Database (Denmark)

    Aanaes, Kasper; Johansen, Helle Krogh; Poulsen, Steen Seier;

    2012-01-01

    BACKGROUND: Pseudomonas aeruginosa sinusitis may be the focus for intermittent lung colonization in patients with cystic fibrosis (CF). The sinusitis may induce elevated IgA levels in nasal secretion and saliva against P. aeruginosa. METHODS: 120 CF patients chronically infected, intermittently...... secretions and saliva than those without P. aeruginosa in the lungs, indicating that P. aeruginosa sinusitis may precede intermittent colonization and chronic infection of the lungs. CONCLUSIONS: Specific IgA against P. aeruginosa in nasal secretions and saliva can contribute to differentiation between...

  15. Polysaccharides serve as scaffold of biofilms formed by mucoid Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Yang, Liang; Hengzhuang, Wang; Wu, Hong

    2012-01-01

    Chronic lung infection by mucoid Pseudomonas aeruginosa is one of the major pathologic features in patients with cystic fibrosis. Mucoid P. aeruginosa is notorious for its biofilm forming capability and resistance to immune attacks. In this study, the roles of extracellular polymeric substances...... from biofilms formed by mucoid P. aeruginosa were investigated. Alginate is not an essential structure component for mucoid P. aeruginosa biofilms. Genetic studies revealed that Pel and Psl polysaccharides serve as essential scaffold and mediate macrocolony formation in mucoid P. aeruginosa biofilms...

  16. Accelerated corrosion of 2205 duplex stainless steel caused by marine aerobic Pseudomonas aeruginosa biofilm.

    Science.gov (United States)

    Xu, Dake; Xia, Jin; Zhou, Enze; Zhang, Dawei; Li, Huabing; Yang, Chunguang; Li, Qi; Lin, Hai; Li, Xiaogang; Yang, Ke

    2017-02-01

    Microbiologically influenced corrosion (MIC) of 2205 duplex stainless steel (DSS) in the presence of Pseudomonas aeruginosa was investigated through electrochemical and surface analyses. The electrochemical results showed that P. aeruginosa significantly reduced the corrosion resistance of 2205 DSS. Confocal laser scanning microscopy (CLSM) images showed that the depths of the largest pits on 2205 DSS with and without P. aeruginosa were 14.0 and 4.9μm, respectively, indicating that the pitting corrosion was accelerated by P. aeruginosa. X-ray photoelectron spectroscopy (XPS) results revealed that CrO3 and CrN formed on the 2205 DSS surface in the presence of P. aeruginosa.

  17. BOX-BEHNKEN EXPERIMENTAL DESIGN MEDIATED OPTIMIZATION OF AQUEOUS METHYLPARATHION BIODEGRADATION BY Pseudomonas aeruginosa mpd STRAIN

    Directory of Open Access Journals (Sweden)

    Krishnaswamy Usharani

    2016-06-01

    Full Text Available Biotreatment of methylparathion was studied in aqueous mineral salts medium containing bacterial culture to demonstrate the potential of the novel strain of Pseudomonas aeruginosa mpd. A statistical Box–Behnken Design (BBD of experiments was performed to evaluate the effects of individual operating variables and their interactions on the methylparathion removal with initial concentration of 1000 mg l-1 as fixed input parameter. The temperature (X1, pH (X2, reaction time (X3 and agitation (X4 were used as design factors. The result was shown that experimental data fitted with the polynomial model. Analysis of variance showed a high coefficient of determination value 0.9. The optimum biodegradation of MP in terms MP removal (Y1, COD removal (Y2 and TOC removal (Y3 were found to be 95.2 %, 82 % and 61.2 % respectively. The maximum growth (Y4 was 2.18 optical density (OD. The optimum biodegradation correspond to the factors combination of middle level of X1 (33 oC, X2 (7.0, X4 (150 rpm and the highest level of X3 (96h. MP removal and its residues were detected using spectral analysis. The study demonstrates the optimum MP biodegradation potential of this strain could use MP as the sole Carbon/Phosphate source. BBD confirmed to be dependable in developing the model, optimizing factors and analyzing interaction effects. Data from this study will be helpful in the design of small-scale field experiments and subsequently an in situ methylparathion biotreatment system for field application.

  18. Antibodies against Pseudomonas aeruginosa chromosomal beta-lactamase inpatients with cystic fibrosis are markers of the development of resistance of P. aeruginosa to beta-lactams

    DEFF Research Database (Denmark)

    Ciofu, O; Giwercman, B; Walter-Rasmussen, J

    1995-01-01

    Chromosomal beta-lactamase production is considered to be the most important resistance mechanism of Pseudomonas aeruginosa against beta-lactams. Recently we have detected serum and sputum antibodies against P. aeruginosa chromosomal beta-lactamase (a beta ab), using immunoblotting techniques...... infection and was significantly higher (P beta-lactam courses. A 14 fold increase in a beta ab...... levels occurred during the 14 year period covered by the longitudinal study. The results of this study show that a beta ab to P. aeruginosa is a specific marker for resistance development of P. aeruginosa to beta-lactams....

  19. Pseudomonas aeruginosa facilitates Campylobacter jejuni growth in biofilms under oxic flow conditions.

    Science.gov (United States)

    Culotti, Alessandro; Packman, Aaron I

    2015-12-01

    We investigated the growth of Campylobacter jejuni in biofilms with Pseudomonas aeruginosa under oxic flow conditions. We observed the growth of C. jejuni in mono-culture, deposited on pre-established P. aeruginosa biofilms, and co-inoculated with P. aeruginosa. In mono-culture, C. jejuni was unable to form biofilms. However, deposited C. jejuni continuously grew on pre-established P. aeruginosa biofilms for a period of 3 days. The growth of scattered C. jejuni clusters was strictly limited to the P. aeruginosa biofilm surface, and no intergrowth was observed. Co-culturing of C. jejuni and P. aeruginosa also enabled the growth of both organisms in biofilms, with C. jejuni clusters developing on the surface of the P. aeruginosa biofilm. Dissolved oxygen (DO) measurements in the medium showed that P. aeruginosa biofilms depleted the effluent DO from 9.0 to 0.5 mg L(-1) 24 hours after inoculation. The localized microaerophilic environment generated by P. aeruginosa promoted the persistence and growth of C. jejuni. Our findings show that P. aeruginosa not only prolongs the survival of C. jejuni under oxic conditions, but also enables the growth of C. jejuni on the surface of P. aeruginosa biofilms.

  20. Pseudomonas aeruginosa promotes Escherichia coli biofilm formation in nutrient-limited medium.

    Directory of Open Access Journals (Sweden)

    Alessandro Culotti

    Full Text Available Biofilms have been implicated as an important reservoir for pathogens and commensal enteric bacteria such as Escherichia coli in natural and engineered water systems. However, the processes that regulate the survival of E. coli in aquatic biofilms have not been thoroughly studied. We examined the effects of hydrodynamic shear and nutrient concentrations on E. coli colonization of pre-established Pseudomonas aeruginosa biofilms, co-inoculation of E. coli and P. aeruginosa biofilms, and P. aeruginosa colonization of pre-established E. coli biofilms. In nutritionally-limited R2A medium, E. coli dominated biofilms when co-inoculated with P. aeruginosa, and successfully colonized and overgrew pre-established P. aeruginosa biofilms. In more enriched media, P. aeruginosa formed larger clusters, but E. coli still extensively overgrew and colonized the interior of P. aeruginosa clusters. In mono-culture, E. coli formed sparse and discontinuous biofilms. After P. aeruginosa was introduced to these biofilms, E. coli growth increased substantially, resulting in patterns of biofilm colonization similar to those observed under other sequences of organism introduction, i.e., E. coli overgrew P. aeruginosa and colonized the interior of P. aeruginosa clusters. These results demonstrate that E. coli not only persists in aquatic biofilms under depleted nutritional conditions, but interactions with P. aeruginosa can greatly increase E. coli growth in biofilms under these experimental conditions.

  1. Interactions between the antimicrobial agent triclosan and the bloom-forming cyanobacteria Microcystis aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Xiaolong [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Wuhan Zhongke Hydrobiological Environment Engineering Co., Ltd, Wuhan 430071 (China); Tu, Yenan; Song, Chaofeng; Li, Tiancui; Lin, Juan [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Graduate University of the Chinese Academy of Sciences, Beijing 100049 (China); Wu, Yonghong [State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Sciences, Chinese Academy of Sciences, Nanjing 210008 (China); Liu, Jiantong [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Wu, Chenxi, E-mail: chenxi.wu@ihb.ac.cn [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China)

    2016-03-15

    Highlights: • Triclosan inhibit the growth and photosynthesis of M. aeruginosa at environmental relevant level. • TEM imaging showed destruction of M. aeruginosa cell ultrastructure during triclosan exposure. • Triclosan can be biotransformed by M. aeruginosa with methylation as a major pathway. • Presence of M. aeruginosa enhanced the photodegradation of triclosan. - Abstract: Cyanobacteria can co-exist in eutrophic waters with chemicals or other substances derived from personal care products discharged in wastewater. In this work, we investigate the interactions between the antimicrobial agent triclosan (TCS) and the bloom-forming cyanobacteria Microcystis aeruginosa. M. aeruginosa was very sensitive to TCS with the 96 h lowest observed effect concentration of 1.0 and 10 μg/L for inhibition of growth and photosynthetic activity, respectively. Exposure to TCS at environmentally relevant levels (0.1–2.0 μg/L) also affected the activities of superoxide dismutase (SOD) and the generation of reduced glutathione (GSH), while microcystin production was not affected. Transmission electron microscope (TEM) examination showed the destruction of M. aeruginosa cell ultrastructure during TCS exposure. TCS however, can be biotransformed by M. aeruginosa with methylation as a major biotransformation pathway. Furthermore, the presence of M. aeruginosa in solution promoted the photodegradation of TCS. Overall, our results demonstrate that M. aeruginosa plays an important role in the dissipation of TCS in aquatic environments but high residual TCS can exert toxic effects on M. aeruginosa.

  2. Zingerone silences quorum sensing and attenuates virulence of Pseudomonas aeruginosa.

    Science.gov (United States)

    Kumar, Lokender; Chhibber, Sanjay; Kumar, Rajnish; Kumar, Manoj; Harjai, Kusum

    2015-04-01

    Quorum sensing in Pseudomonas aeruginosa plays an imperative role in virulence factor, biofilm formation and antimicrobial resistance. Blocking quorum sensing pathways are viewed as viable anti-virulent therapy in association with traditional antimicrobial therapy. Anti-quorum sensing dietary phytochemicals with may prove to be a safe and viable choice as anti-virulent drug candidates. Previously, our lab proved zingerone as potent anti-biofilm agent hence; further its anti-virulent and anti-quorum activities were evaluated. Zingerone, besides decreasing swimming, swarming and twitching phenotypes of P. aeruginosa PAO1, reduced biofilm forming capacity and production of virulence factors including rhamnolipid, elastase, protease, pyocyanin, cell free and cell bound hemolysin (pquorum sensing signal molecules by clinical isolates of P. aeruginosa but also showed significant interference with the activation of QS reporter strains. To study the mechanism of blocking quorum sensing cascade, in silico analysis was carried out. Anti-QS activity was attributed to interference with the ligand receptor interaction of zingerone with QS receptors (TraR, LasR, RhlR and PqsR). Zingerone showed a good comparative docking score to respective autoinducer molecules which was even higher than that of vanillin, a proven anti-quorum sensing phytochemical. The results of the present study revealed the anti-quorum sensing activity of zingerone targeting ligand-receptor interaction, hence proposing zingerone as a suitable anti-virulent drug candidate against P. aeruginosa infections.

  3. Ciprofloxacin susceptibility of Pseudomonas aeruginosa isolates from keratitis

    DEFF Research Database (Denmark)

    Lomholt, JA; Kilian, Mogens

    2003-01-01

    AIM: To examine the ciprofloxacin susceptibility of 106 Pseudomonas aeruginosa eye isolates from the United Kingdom, Denmark, India, the United States, and Australia, and to determine the molecular mechanisms of resistance. METHODS: Ciprofloxacin susceptibility was tested by an agar dilution meth...

  4. AMINOGLYCOSIDE RESISTANCE GENES IN Pseudomonas aeruginosa ISOLATES FROM CUMANA, VENEZUELA

    Science.gov (United States)

    TEIXEIRA, Bertinellys; RODULFO, Hectorina; CARREÑO, Numirin; GUZMÁN, Militza; SALAZAR, Elsa; DONATO, Marcos DE

    2016-01-01

    The enzymatic modification of aminoglycosides by aminoglycoside-acetyltransferases (AAC), aminoglycoside-adenyltransferases (AAD), and aminoglycoside-phosphotransferases (APH), is the most common resistance mechanism in P. aeruginosa and these enzymes can be coded on mobile genetic elements that contribute to their dispersion. One hundred and thirty seven P. aeruginosa isolates from the University Hospital, Cumana, Venezuela (HUAPA) were evaluated. Antimicrobial susceptibility was determined by the disk diffusion method and theaac, aadB and aph genes were detected by PCR. Most of the P. aeruginosa isolates (33/137) were identified from the Intensive Care Unit (ICU), mainly from discharges (96/137). The frequency of resistant P. aeruginosaisolates was found to be higher for the aminoglycosides tobramycin and amikacin (30.7 and 29.9%, respectively). Phenotype VI, resistant to these antibiotics, was the most frequent (14/49), followed by phenotype I, resistant to all the aminoglycosides tested (12/49). The aac(6´)-Ib,aphA1 and aadB genes were the most frequently detected, and the simultaneous presence of several resistance genes in the same isolate was demonstrated. Aminoglycoside resistance in isolates ofP. aeruginosa at the HUAPA is partly due to the presence of the aac(6´)-Ib, aphA1 andaadB genes, but the high rates of antimicrobial resistance suggest the existence of several mechanisms acting together. This is the first report of aminoglycoside resistance genes in Venezuela and one of the few in Latin America. PMID:27007556

  5. Reduction of PCN biosynthesis by NO in Pseudomonas aeruginosa.

    Science.gov (United States)

    Gao, Lei; Zhang, Yuying; Wang, Yan; Qiao, Xinhua; Zi, Jing; Chen, Chang; Wan, Yi

    2016-08-01

    Pyocyanin (PCN), a virulence factor synthesized by Pseudomonas aeruginosa, plays an important role during clinical infections. There is no study of the effect of nitric oxide (NO) on PCN biosynthesis. Here, the effect of NO on PCN levels in Pseudomonas aeruginosa strain PAO1, a common reference strain, was tested. The results showed that the NO donor sodium nitroprusside (SNP) can significantly reduce PCN levels (82.5% reduction at 60μM SNP). Furthermore, the effect of endogenous NO on PCN was tested by constructing PAO1 nor (NO reductase gene) knockout mutants. Compared to the wild-type strain, the Δnor strain had a lower PCN (86% reduction in Δnor). To examine whether the results were universal with other P. aeruginosa strains, we collected 4 clinical strains from a hospital, tested their PCN levels after SNP treatment, and obtained similar results, i.e., PCN biosynthesis was inhibited by NO. These results suggest that NO treatment may be a new strategy to inhibit PCN biosynthesis and could provide novel insights into eliminating P. aeruginosa virulence as a clinical goal.

  6. Reduction of PCN biosynthesis by NO in Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Lei Gao

    2016-08-01

    Full Text Available Pyocyanin (PCN, a virulence factor synthesized by Pseudomonas aeruginosa, plays an important role during clinical infections. There is no study of the effect of nitric oxide (NO on PCN biosynthesis. Here, the effect of NO on PCN levels in Pseudomonas aeruginosa strain PAO1, a common reference strain, was tested. The results showed that the NO donor sodium nitroprusside (SNP can significantly reduce PCN levels (82.5% reduction at 60 μM SNP. Furthermore, the effect of endogenous NO on PCN was tested by constructing PAO1 nor (NO reductase gene knockout mutants. Compared to the wild-type strain, the Δnor strain had a lower PCN (86% reduction in Δnor. To examine whether the results were universal with other P. aeruginosa strains, we collected 4 clinical strains from a hospital, tested their PCN levels after SNP treatment, and obtained similar results, i.e., PCN biosynthesis was inhibited by NO. These results suggest that NO treatment may be a new strategy to inhibit PCN biosynthesis and could provide novel insights into eliminating P. aeruginosa virulence as a clinical goal.

  7. Induction of beta-lactamase production in Pseudomonas aeruginosa biofilm

    DEFF Research Database (Denmark)

    Giwercman, B; Jensen, E T; Høiby, N;

    1991-01-01

    Imipenem induced high levels of beta-lactamase production in Pseudomonas aeruginosa biofilms. Piperacillin also induced beta-lactamase production in these biofilms but to a lesser degree. The combination of beta-lactamase production with other protective properties of the biofilm mode of growth...

  8. Optimized electroporation-induced transformation in Microcystis aeruginosa PCC7806

    Directory of Open Access Journals (Sweden)

    El Semary, A.

    2010-01-01

    Full Text Available Gene disruption in cyanobacteria is difficult and comprises an obstacle for genetic manipulation. Very few reports tackled this problem but the methods used are usually obscure and hardly reproducible. Here we describe an optimized electroporation-induced transformation in Microcystis aeruginosa PCC7806 where conditions for successful electroporation and transformation are investigated.

  9. AMINOGLYCOSIDE RESISTANCE GENES IN Pseudomonas aeruginosa ISOLATES FROM CUMANA, VENEZUELA.

    Science.gov (United States)

    Teixeira, Bertinellys; Rodulfo, Hectorina; Carreño, Numirin; Guzmán, Militza; Salazar, Elsa; De Donato, Marcos

    2016-01-01

    The enzymatic modification of aminoglycosides by aminoglycoside-acetyltransferases (AAC), aminoglycoside-adenyltransferases (AAD), and aminoglycoside-phosphotransferases (APH), is the most common resistance mechanism in P. aeruginosa and these enzymes can be coded on mobile genetic elements that contribute to their dispersion. One hundred and thirty seven P. aeruginosa isolates from the University Hospital, Cumana, Venezuela (HUAPA) were evaluated. Antimicrobial susceptibility was determined by the disk diffusion method and theaac, aadB and aph genes were detected by PCR. Most of the P. aeruginosa isolates (33/137) were identified from the Intensive Care Unit (ICU), mainly from discharges (96/137). The frequency of resistant P. aeruginosaisolates was found to be higher for the aminoglycosides tobramycin and amikacin (30.7 and 29.9%, respectively). Phenotype VI, resistant to these antibiotics, was the most frequent (14/49), followed by phenotype I, resistant to all the aminoglycosides tested (12/49). The aac(6´)-Ib,aphA1 and aadB genes were the most frequently detected, and the simultaneous presence of several resistance genes in the same isolate was demonstrated. Aminoglycoside resistance in isolates ofP. aeruginosa at the HUAPA is partly due to the presence of the aac(6´)-Ib, aphA1 andaadB genes, but the high rates of antimicrobial resistance suggest the existence of several mechanisms acting together. This is the first report of aminoglycoside resistance genes in Venezuela and one of the few in Latin America.

  10. MexXY multidrug efflux system of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Yuji eMorita

    2012-11-01

    Full Text Available Anti-pseudomonas aminoglycosides, such as amikacin and tobramycin, are used in the treatment of Pseudomonas aeruginosa infections. However, their use is linked to the development of resistance. During the last decade, the MexXY multidrug efflux system has been comprehensively studied, and numerous reports of laboratory and clinical isolates have been published. This system has been increasingly recognized as one of the primary determinants of aminoglycoside resistance in P. aeruginosa. In P. aeruginosa cystic fibrosis isolates, upregulation of the pump is considered the most common mechanism of aminoglycoside resistance. Non-fermentative Gram-negative pathogens possessing very close MexXY orthologues such as Achromobacter xylosoxidans and various Burkholderia species [e.g., B. pseudomallei and B. cepacia complexes], but not B. gladioli, are intrinsically resistant to aminoglycosides. Here, we summarize the properties (e.g., discovery, mechanism, gene expression, clinical significance of the P. aeruginosa MexXY pump and other aminoglycoside efflux pumps such as AcrD of Escherichia coli, AmrAB-OprA of B. pseudomallei, and AdeABC of Acinetobacter baumannii. MexXY inducibility of the PA5471 gene product, which is dependent on ribosome inhibition or oxidative stress, is noteworthy. Moreover, the discovery of the cognate outer membrane component (OprA of MexXY in the multidrug-resistant clinical isolate PA7, serotype O12 deserves special attention.

  11. Inhibition of Pseudomonas aeruginosa biofilm formation on wound dressings.

    Science.gov (United States)

    Brandenburg, Kenneth S; Calderon, Diego F; Kierski, Patricia R; Brown, Amanda L; Shah, Nihar M; Abbott, Nicholas L; Schurr, Michael J; Murphy, Christopher J; McAnulty, Jonathan F; Czuprynski, Charles J

    2015-01-01

    Chronic nonhealing skin wounds often contain bacterial biofilms that prevent normal wound healing and closure and present challenges to the use of conventional wound dressings. We investigated inhibition of Pseudomonas aeruginosa biofilm formation, a common pathogen of chronic skin wounds, on a commercially available biological wound dressing. Building on prior reports, we examined whether the amino acid tryptophan would inhibit P. aeruginosa biofilm formation on the three-dimensional surface of the biological dressing. Bacterial biomass and biofilm polysaccharides were quantified using crystal violet staining or an enzyme linked lectin, respectively. Bacterial cells and biofilm matrix adherent to the wound dressing were visualized through scanning electron microscopy. D-/L-tryptophan inhibited P. aeruginosa biofilm formation on the wound dressing in a dose dependent manner and was not directly cytotoxic to immortalized human keratinocytes although there was some reduction in cellular metabolism or enzymatic activity. More importantly, D-/L-tryptophan did not impair wound healing in a splinted skin wound murine model. Furthermore, wound closure was improved when D-/L-tryptophan treated wound dressing with P. aeruginosa biofilms were compared with untreated dressings. These findings indicate that tryptophan may prove useful for integration into wound dressings to inhibit biofilm formation and promote wound healing.

  12. Typing of Pseudomonas aeruginosa strains in Norwegian cystic fibrosis patients

    DEFF Research Database (Denmark)

    Fluge, G; Ojeniyi, B; Høiby, N;

    2001-01-01

    OBJECTIVES: Typing of Pseudomonas aeruginosa isolates from Norwegian cystic fibrosis (CF) patients with chronic Pseudomonas lung infection in order to see whether cross-infection might have occurred. METHODS: Isolates from 60 patients were collected during the years 1994-98, and typed by pulsed...

  13. Pseudomonas aeruginosa diversity in distinct paediatric patient groups

    DEFF Research Database (Denmark)

    Tramper-Stranders, G.A.; Ent, C.K. van der; Wolfs, T.F.;

    2008-01-01

    and further typed by pulsed-field gel electrophoresis. Simpson's diversity index was calculated for the five groups. CF-chronic patients carried the highest number of distinct P. aeruginosa phenotypes and genotypes per culture. Isolates from the CF-chronic group were significantly less diverse than those from...

  14. Maturation of Pseudomonas aeruginosa elastase - Formation of the disulfide bonds

    NARCIS (Netherlands)

    Braun, P; Ockhuijsen, C; Eppens, E; Koster, M; Bitter, W; Tommassen, J

    2001-01-01

    Elastase of Pseudomonas aeruginosa is synthesized as a preproenzyme. After propeptide-mediated folding in the periplasm, the proenzyme is autoproteolytically processed, prior to translocation of both the mature enzyme and the propeptide across the outer membrane. The formation of the two disulfide b

  15. Pyoverdine, the Major Siderophore in Pseudomonas aeruginosa, Evades NGAL Recognition

    Directory of Open Access Journals (Sweden)

    Mary E. Peek

    2012-01-01

    Full Text Available Pseudomonas aeruginosa is the most common pathogen that persists in the cystic fibrosis lungs. Bacteria such as P. aeruginosa secrete siderophores (iron-chelating molecules and the host limits bacterial growth by producing neutrophil-gelatinase-associated lipocalin (NGAL that specifically scavenges bacterial siderophores, therefore preventing bacteria from establishing infection. P. aeruginosa produces a major siderophore known as pyoverdine, found to be important for bacterial virulence and biofilm development. We report that pyoverdine did not bind to NGAL, as measured by tryptophan fluorescence quenching, while enterobactin bound to NGAL effectively causing a strong response. The experimental data indicate that pyoverdine evades NGAL recognition. We then employed a molecular modeling approach to simulate the binding of pyoverdine to human NGAL using NGAL’s published crystal structures. The docking of pyoverdine to NGAL predicted nine different docking positions; however, neither apo- nor ferric forms of pyoverdine docked into the ligand-binding site in the calyx of NGAL where siderophores are known to bind. The molecular modeling results offer structural support that pyoverdine does not bind to NGAL, confirming the results obtained in the tryptophan quenching assay. The data suggest that pyoverdine is a stealth siderophore that evades NGAL recognition allowing P. aeruginosa to establish chronic infections in CF lungs.

  16. Role of mutation in Pseudomonas aeruginosa biofilm development.

    Directory of Open Access Journals (Sweden)

    Tim C R Conibear

    Full Text Available The survival of bacteria in nature is greatly enhanced by their ability to grow within surface-associated communities called biofilms. Commonly, biofilms generate proliferations of bacterial cells, called microcolonies, which are highly recalcitrant, 3-dimensional foci of bacterial growth. Microcolony growth is initiated by only a subpopulation of bacteria within biofilms, but processes responsible for this differentiation remain poorly understood. Under conditions of crowding and intense competition between bacteria within biofilms, microevolutionary processes such as mutation selection may be important for growth; however their influence on microcolony-based biofilm growth and architecture have not previously been explored. To study mutation in-situ within biofilms, we transformed Pseudomonas aeruginosa cells with a green fluorescent protein gene containing a +1 frameshift mutation. Transformed P. aeruginosa cells were non-fluorescent until a mutation causing reversion to the wildtype sequence occurs. Fluorescence-inducing mutations were observed in microcolony structures, but not in other biofilm cells, or in planktonic cultures of P. aeruginosa cells. Thus microcolonies may represent important foci for mutation and evolution within biofilms. We calculated that microcolony-specific increases in mutation frequency were at least 100-fold compared with planktonically grown cultures. We also observed that mutator phenotypes can enhance microcolony-based growth of P. aeruginosa cells. For P. aeruginosa strains defective in DNA fidelity and error repair, we found that microcolony initiation and growth was enhanced with increased mutation frequency of the organism. We suggest that microcolony-based growth can involve mutation and subsequent selection of mutants better adapted to grow on surfaces within crowded-cell environments. This model for biofilm growth is analogous to mutation selection that occurs during neoplastic progression and tumor

  17. Imipenem-resistant Pseudomonas aeruginosa: risk factors for nosocomial infections.

    Science.gov (United States)

    Onguru, Pinar; Erbay, Ayse; Bodur, Hurrem; Baran, Gulseren; Akinci, Esragul; Balaban, Neriman; Cevik, Mustafa Aydin

    2008-12-01

    The aim of this study was to determine the risk factors for nosocomial infections of imipenem-resistant Pseudomonas aeruginosa (IRPA). A prospective case-control study was performed at a tertiary care hospital in Ankara from January to December 2004. The patients with nosocomial P. aeruginosa infection were included in the study. The features of the patients with IRPA infections were compared to those with imipenem-sensitive P. aeruginosa (ISPA) infections. Only the first isolation of P. aeruginosa was considered. Nosocomial infections were defined according to Center for Disease Control (CDC) criteria. IRPA was isolated from 75 (44.1%) patients, and ISPA was isolated from 95 (55.9%) patients during the study period. IRPA were most frequently isolated from endotracheal aspirate (19%) cultures (p=0.048), whereas ISPA were most frequently isolated from urine (28%) cultures (p=0.023). In multivariate analysis, a longer duration of hospital stay until P. aeruginosa isolation (odds ratio [OR], 1.027; 95% confidence interval [CI], 1.002-1.054, p=0.034), arterial catheter administration (OR, 2.508; 95% CI, 1.062-5.920, p=0.036), vancomycin (OR, 2.882; 95% CI, 1.130-7.349, p=0.027), piperacillin-tazobactam (OR, 6.425; 95% CI, 2.187-18.875, p=0.001), and imipenem (OR, 3.580; 95% CI, 1.252-10.245, p=0.017) treatment within the 14 days before isolation of IRPA were independently associated with imipenem resistance. It was concluded that treatment with imipenem, vancomycin and piperacillin-tazobactam were major risk factors for IRPA infections in hospitalized patients. The nosocomial occurrence of IRPA was also strongly related to the duration of hospital stay, arterial catheter administration.

  18. Periplasmic response upon disruption of transmembrane Cu transport in Pseudomonas aeruginosa.

    Science.gov (United States)

    Raimunda, Daniel; Padilla-Benavides, Teresita; Vogt, Stefan; Boutigny, Sylvain; Tomkinson, Kaleigh N; Finney, Lydia A; Argüello, José M

    2013-02-01

    Pseudomonas aeruginosa, an opportunistic pathogen, has two transmembrane Cu(+) transport ATPases, CopA1 and CopA2. Both proteins export cytoplasmic Cu(+) into the periplasm and mutation of either gene leads to attenuation of virulence. CopA1 is required for maintaining cytoplasmic copper levels, while CopA2 provides copper for cytochrome c oxidase assembly. We hypothesized that transported Cu(+) ions would be directed to their destination via specific periplasmic partners and disruption of transport should affect the periplasmic copper homeostasis. Supporting this, mutation of either ATPase gene led to large increments in periplasmic cuproprotein levels. Toward identifying the proteins participating in this cellular response the periplasmic metalloproteome was resolved in non-denaturing bidimensional gel electrophoresis, followed by X-ray fluorescence visualization and identification by mass-spectrometry. A single spot containing the electron shuttle protein azurin was responsible for the observed increments in cuproprotein contents. In agreement, lack of either Cu(+)-ATPase induced an increase in azu transcription. This is associated with an increase in the expression of anr and rpoS oxidative stress response regulators, rather than cueR, a copper sensing regulator. We propose that azurin overexpression and accumulation in the periplasm is part of the cellular response to cytoplasmic oxidative stress in P. aeruginosa.

  19. Bdellovibrio bacteriovorus directly attacks Pseudomonas aeruginosa and Staphylococcus aureus Cystic fibrosis isolates

    Directory of Open Access Journals (Sweden)

    Valerio eIebba

    2014-06-01

    Full Text Available Bdellovibrio bacteriovorus is a predator bacterial species found in the environment and within the human gut, able to attack Gram-negative prey. Cystic fibrosis (CF is a genetic disease which usually presents lung colonization by Pseudomonas aeruginosa or Staphylococcus aureus biofilms. Here, we investigated the predatory behaviour of B. bacteriovorus against these two pathogenic species with: i broth culture; ii ‘static’ biofilms; iii field emission scanning electron microscope (FESEM; iv ‘flow’ biofilms; v zymographic technique. We had the first evidence of B. bacteriovorus survival with a Gram-positive prey, revealing a direct cell-to-cell contact with S. aureus and a new ‘epibiotic’ foraging strategy imaged with FESEM. Mean attaching time of HD100 to S. aureus cells was 185s, while ‘static’ and ‘flow’ S. aureus biofilms were reduced by 74% (at 24h and 46% (at 20h, respectively. Furthermore, zymograms showed a differential bacteriolytic activity exerted by the B. bacteriovorus lysates on P. aeruginosa and S. aureus. The dual foraging system against Gram-negative (periplasmic and Gram-positive (epibiotic prey could suggest the use of B. bacteriovorus as a ‘living antibiotic’ in CF, even if further studies are required to simulate its in vivo predatory behaviour.

  20. An orphan chemotaxis sensor regulates virulence and antibiotic tolerance in the human pathogen Pseudomonas aeruginosa.

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    Heather Pearl McLaughlin

    Full Text Available The synthesis of virulence factors by pathogenic bacteria is highly regulated and occurs in response to diverse environmental cues. An array of two component systems (TCSs serves to link perception of different cues to specific changes in gene expression and/or bacterial behaviour. Those TCSs that regulate functions associated with virulence represent attractive targets for interference in anti-infective strategies for disease control. We have previously identified PA2572 as a putative response regulator required for full virulence of Pseudomonas aeruginosa, the opportunistic human pathogen, to Galleria mellonella (Wax moth larvae. Here we have investigated the involvement of candidate sensors for signal transduction involving PA2572. Mutation of PA2573, encoding a probable methyl-accepting chemotaxis protein, gave rise to alterations in motility, virulence, and antibiotic resistance, functions which are also controlled by PA2572. Comparative transcriptome profiling of mutants revealed that PA2572 and PA2573 regulate expression of a common set of 49 genes that are involved in a range of biological functions including virulence and antibiotic resistance. Bacterial two-hybrid analysis indicated a REC-dependent interaction between PA2572 and PA2573 proteins. Finally expression of PA2572 in the PA2573 mutant background restored virulence to G. mellonella towards wild-type levels. The findings indicate a role for the orphan chemotaxis sensor PA2573 in the regulation of virulence and antibiotic tolerance in P. aeruginosa and indicate that these effects are exerted in part through signal transduction involving PA2572.

  1. Orf1/SpcS Chaperones ExoS for Type Three Secretion by Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    DA-KANG SHEN; LAURIANE QUENEE; MARIETTE BONNET; LAURIANE KUHN; MADIHA DEROUAZI; DANIELE LAMOTTE; BERTRAND TOUSSAINT; BENOIT POLACK

    2008-01-01

    Objective Pseudomonas aeruginosa is a ubiquitous and opportunistic pathogen that uses the type III secretion system (TTSS)to inject effector proteins directly into the cytosol of target cells to subvert the host cell's functions.Specialized bacterial chaperones are required for effective secretion of some effectors.To identify the chaperone of ExoS,the representative effector secreted by the TTSS of P. aeruginosa,we analyzed the role of a postulated chaperone termed Orfl.Methods By allelic exchange,we constructed the mutant with the deletion of gene Orfl.Analysis of secreted and cell-associated fractions was performed by SDS-PAGE and Western blotting.Using strain expressing in trans Orfl,tagged by V5 polypeptide and histidine,protein-protein interaction Was determined by affinity resin pull-down assay in combination with MALDI-TOF. The role of Orfl in the expression of eroS was evaluated by genel reporter analysis.Results Pull-down assay showed that Orfl binds to ExoS and ExoT.Secretion profile analysis showedthat Orfl was necessary for the optimal secretion of ExoS and ExoT.However,Orfl had no effect on the expression of eroS.Conclusion Orfl is important for the secretion of ExoS probablY by mamtauung ExoS in a secretion-competent conformation.We propose to name Orfl as SpcS for "specific Pseudomonas chaperone for ExoS".

  2. Inactivation of Pseudomonas aeruginosa in electrochemical advanced oxidation process with diamond electrodes.

    Science.gov (United States)

    Griessler, M; Knetsch, S; Schimpf, E; Schmidhuber, A; Schrammel, B; Wesner, W; Sommer, R; Kirschner, A K T

    2011-01-01

    The electrochemical advanced oxidation process (EAOP) with diamond electrodes may serve as an additional technology to the currently approved methods for water disinfection. Only few data exist on the microbicidal effect of the EAOP. The aim of our study was to investigate the microbicidal effect of a flow-through oxidation cell with diamond electrodes, using Pseudomonas aeruginosa as the test organism. Without electrical current the EAOP had no measurable effect on investigated microbiological and chemical parameters. For direct electrical current a stronger impact was observed at low flow rate than at higher flow rate. Depending on the contact time of the oxidants and the type of quenching reagent added, inactivation of P. aeruginosa was in the range log 1.6-3.6 at the higher flow rate and log 2.4-4.4 at the lower rate. Direct electrical current showed a stronger microbicidal effect than alternating current (maximum reduction log 4.0 and log 2.9, respectively). The microbiological results of experiments with this EAOP prototype revealed higher standard deviations than expected, based on our experience with standard water disinfection methods. Safe use of an EAOP system requires operating parameters to be defined and used accurately, and thus specific monitoring tests must be developed.

  3. Chlorine disinfection of Pseudomonas aeruginosa, total coliforms, Escherichia coli and Enterococcus faecalis: revisiting reclaimed water regulations.

    Science.gov (United States)

    Coronel-Olivares, Claudia; Reyes-Gómez, Lidia María; Hernández-Muñoz, Aurelio; Martínez-Falcón, Ana Paola; Vázquez-Rodríguez, Gabriela A; Iturbe, Ulises

    2011-01-01

    Pathogenic organisms can be transmitted orally through drinking water or through skin and mucosae by both direct and indirect contact, and their presence in water thus has a negative impact on public health. In wastewater treatment plants (WWTP), water is disinfected to inactivate pathogens. The quantification of several microbial indicators in aquatic systems is required to estimate the biological quality of such systems. So far, coliform bacteria have been used as traditional indicators world-wide. This study has assessed the resistance of total coliforms, Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis to three dosages of sodium hypochlorite (NaClO) at two exposure times. The bacteria were isolated from secondary effluents of a WWTP located in Hidalgo, Mexico. The results show that the number of colony-forming units of all studied bacterial types decreased when both the NaClO concentration and exposure times increased. However, they were not eliminated. The inclusion of the species Pseudomonas aeruginosa in regulations for treated wastewater quality as a new indicator is highly recommended due to its importance as an opportunistic pathogen. The detection of this species along with the traditional organisms could be particulary significant for reclaimed water to be used with direct human contact.

  4. Kinetic modeling of rhamnolipid production by Pseudomonas aeruginosa PAO1 including cell density-dependent regulation.

    Science.gov (United States)

    Henkel, Marius; Schmidberger, Anke; Vogelbacher, Markus; Kühnert, Christian; Beuker, Janina; Bernard, Thomas; Schwartz, Thomas; Syldatk, Christoph; Hausmann, Rudolf

    2014-08-01

    The production of rhamnolipid biosurfactants by Pseudomonas aeruginosa is under complex control of a quorum sensing-dependent regulatory network. Due to a lack of understanding of the kinetics applicable to the process and relevant interrelations of variables, current processes for rhamnolipid production are based on heuristic approaches. To systematically establish a knowledge-based process for rhamnolipid production, a deeper understanding of the time-course and coupling of process variables is required. By combining reaction kinetics, stoichiometry, and experimental data, a process model for rhamnolipid production with P. aeruginosa PAO1 on sunflower oil was developed as a system of coupled ordinary differential equations (ODEs). In addition, cell density-based quorum sensing dynamics were included in the model. The model comprises a total of 36 parameters, 14 of which are yield coefficients and 7 of which are substrate affinity and inhibition constants. Of all 36 parameters, 30 were derived from dedicated experimental results, literature, and databases and 6 of them were used as fitting parameters. The model is able to describe data on biomass growth, substrates, and products obtained from a reference batch process and other validation scenarios. The model presented describes the time-course and interrelation of biomass, relevant substrates, and products on a process level while including a kinetic representation of cell density-dependent regulatory mechanisms.

  5. Kinetics of cell lysis for Microcystis aeruginosa and Nitzschia palea in the exposure to {beta}-cyclocitral

    Energy Technology Data Exchange (ETDEWEB)

    Chang, De-Wei; Hsieh, Meng-Ling [Department of Environmental Engineering, National Cheng Kung University, Tainan City 70101, Taiwan (China); Chen, Yan-Min [Sustainable Environment Research Center, National Cheng Kung University, Tainan City 70101, Taiwan (China); Lin, Tsair-Fuh, E-mail: tflin@mail.ncku.edu.tw [Department of Environmental Engineering, National Cheng Kung University, Tainan City 70101, Taiwan (China); Chang, Jo-Shu [Department of Chemical Engineering, National Cheng Kung University, Tainan City 70101, Taiwan (China)

    2011-01-30

    The effect of an algal metabolite, {beta}-cyclocitral, on the cell integrity of two cyanobacteria and one diatom was investigated. The cyanobacteria, Microcystis aeruginosa PCC 7005 and PCC 7820, and the diatom, Nitzschia palea, were exposed to various concentrations of {beta}-cyclocitral. Scanning electron microscope (SEM) results indicate that the cells of tested species were greatly altered after being exposed to {beta}-cyclocitral. A flow cytometer coupled with the SYTOX stain and chlorophyll-a auto-fluorescence was used to quantify the effect of {beta}-cyclocitral on cell integrity for the tested cyanobacteria and diatom. Kinetic experiments show that about 5-10 mg L{sup -1} of {beta}-cyclocitral for the two M. aeruginosa strains and a much lower concentration, 0.1-0.5 mg L{sup -1}, for N. palea were needed to cause 15-20% of cells to rupture. When the {beta}-cyclocitral concentration was increased to 200-1000 mg L{sup -1} for M. aeruginosa and 5-10 mg L{sup -1} for N. palea, almost all the cells ruptured between 8 and 24 h. A first-order kinetic model is able to describe the data of cell integrity over time. The extracted rate constant values well correlate with the applied {beta}-cyclocitral dosages. The obtained kinetic parameters may be used to estimate {beta}-cyclocitral dosage and contact time required for the control of cyanobacteria and diatoms in water bodies.

  6. Pseudomonas aeruginosa suppresses interferon response to rhinovirus infection in cystic fibrosis but not in normal bronchial epithelial cells.

    Science.gov (United States)

    Chattoraj, Sangbrita S; Ganesan, Shyamala; Faris, Andrea; Comstock, Adam; Lee, Wai-Ming; Sajjan, Umadevi S

    2011-10-01

    Despite increased morbidity associated with secondary respiratory viral infections in cystic fibrosis (CF) patients with chronic Pseudomonas aeruginosa infection, the underlying mechanisms are not well understood. Here, we investigated the effect of P. aeruginosa infection on the innate immune responses of bronchial epithelial cells to rhinovirus (RV) infection. CF cells sequentially infected with mucoid P. aeruginosa (MPA) and RV showed lower levels of interferons (IFNs) and higher viral loads than those of RV-infected cells. Unlike results for CF cells, normal bronchial epithelial cells coinfected with MPA/RV showed higher IFN expression than RV-infected cells. In both CF and normal cells, the RV-stimulated IFN response requires phosphorylation of Akt and interferon response factor 3 (IRF3). Preinfection with MPA inhibited RV-stimulated Akt phosphorylation and decreased IRF3 phosphorylation in CF cells but not in normal cells. Compared to normal, unstimulated CF cells or normal cells treated with CFTR inhibitor showed increased reactive oxygen species (ROS) production. Treatment of CF cells with antioxidants prior to MPA infection partially reversed the suppressive effect of MPA on the RV-stimulated IFN response. Together, these results suggest that MPA preinfection inhibits viral clearance by suppressing the antiviral response particularly in CF cells but not in normal cells. Further, increased oxidative stress in CF cells appears to modulate the innate immune responses to coinfection.

  7. Effect of Cinnamon Oil on Quorum Sensing-Controlled Virulence Factors and Biofilm Formation in Pseudomonas aeruginosa.

    Science.gov (United States)

    Kalia, Manmohit; Yadav, Vivek Kumar; Singh, Pradeep Kumar; Sharma, Deepmala; Pandey, Himanshu; Narvi, Shahid Suhail; Agarwal, Vishnu

    2015-01-01

    Quorum sensing (QS) is a system of stimuli and responses in bacterial cells governed by their population density, through which they regulate genes that control virulence factors and biofilm formation. Despite considerable research on QS and the discovery of new antibiotics, QS-controlled biofilm formation by microorganisms in clinical settings has remained a problem because of nascent drug resistance, which requires screening of diverse compounds for anti-QS activities. Cinnamon is a dietary phytochemical that is traditionally used to remedy digestive problems and assorted contagions, which suggests that cinnamon might contain chemicals that can hinder the QS process. To test this hypothesis, the anti-QS activity of cinnamon oil against P. aeruginosa was tested, measured by the inhibition of biofilm formation and other QS-associated phenomena, including virulence factors such as pyocyanin, rhamnolipid, protease, alginate production, and swarming activity. To this end, multiple microscopy analyses, including light, scanning electron and confocal microscopy, revealed the ability of cinnamon oil to inhibit P. aeruginosa PAO1 biofilms and their accompanying extracellular polymeric substances. This work is the first to demonstrate that cinnamon oil can influence various QS-based phenomena in P. aeruginosa PAO1, including biofilm formation.

  8. Pseudomonas aeruginosa Transmigrates at Epithelial Cell-Cell Junctions, Exploiting Sites of Cell Division and Senescent Cell Extrusion.

    Directory of Open Access Journals (Sweden)

    Guillaume Golovkine

    2016-01-01

    Full Text Available To achieve systemic infection, bacterial pathogens must overcome the critical and challenging step of transmigration across epithelial barriers. This is particularly true for opportunistic pathogens such as Pseudomonas aeruginosa, an agent which causes nosocomial infections. Despite extensive study, details on the mechanisms used by this bacterium to transmigrate across epithelial tissues, as well as the entry sites it uses, remain speculative. Here, using real-time microscopy and a model epithelial barrier, we show that P. aeruginosa employs a paracellular transmigration route, taking advantage of altered cell-cell junctions at sites of cell division or when senescent cells are expelled from the cell layer. Once a bacterium transmigrates, it is followed by a cohort of bacteria using the same entry point. The basal compartment is then invaded radially from the initial penetration site. Effective transmigration and propagation require type 4 pili, the type 3 secretion system (T3SS and a flagellum, although flagellum-deficient bacteria can occasionally invade the basal compartment from wounded areas. In the basal compartment, the bacteria inject the T3SS toxins into host cells, disrupting the cytoskeleton and focal contacts to allow their progression under the cells. Thus, P. aeruginosa exploits intrinsic host cell processes to breach the epithelium and invade the subcellular compartment.

  9. A three-phase in-vitro system for studying Pseudomonas aeruginosa adhesion and biofilm formation upon hydrogel contact lenses

    Directory of Open Access Journals (Sweden)

    Kohlmann Thomas

    2010-11-01

    Full Text Available Abstract Background Pseudomonas aeruginosa is commonly associated with contact lens (CL -related eye infections, for which bacterial adhesion and biofilm formation upon hydrogel CLs is a specific risk factor. Whilst P. aeruginosa has been widely used as a model organism for initial biofilm formation on CLs, in-vitro models that closely reproduce in-vivo conditions have rarely been presented. Results In the current investigation, a novel in-vitro biofilm model for studying the adherence of P. aeruginosa to hydrogel CLs was established. Nutritional and interfacial conditions similar to those in the eye of a CL wearer were created through the involvement of a solid:liquid and a solid:air interface, shear forces and a complex artificial tear fluid. Bioburdens varied depending on the CL material and biofilm maturation occurred after 72 h incubation. Whilst a range of biofilm morphologies were visualised including dispersed and adherent bacterial cells, aggregates and colonies embedded in extracellular polymer substances (EPS, EPS fibres, mushroom-like formations, and crystalline structures, a compact and heterogeneous biofilm morphology predominated on all CL materials. Conclusions In order to better understand the process of biofilm formation on CLs and to test the efficacy of CL care solutions, representative in-vitro biofilm models are required. Here, we present a three-phase biofilm model that simulates the environment in the eye of a CL wearer and thus generates biofilms which resemble those commonly observed in-situ.

  10. Plasma-mediated inactivation of Pseudomonas aeruginosa biofilms grown on borosilicate surfaces under continuous culture system.

    Science.gov (United States)

    Vandervoort, Kurt G; Brelles-Mariño, Graciela

    2014-01-01

    Biofilms are microbial communities attached to a surface and embedded in a matrix composed of exopolysaccharides and excreted nucleic acids. Bacterial biofilms are responsible for undesirable effects such as disease, prostheses colonization, biofouling, equipment damage, and pipe plugging. Biofilms are also more resilient than free-living cells to regular sterilization methods and therefore it is indispensable to develop better ways to control and remove them. The use of gas discharge plasmas is a good alternative since plasmas contain a mixture of reactive agents well-known for their decontamination potential against free microorganisms. We have previously reported that Pseudomonas aeruginosa biofilms were inactivated after a 1-min plasma exposure. We determined that the adhesiveness and the thickness of Pseudomonas biofilms grown on borosilicate were reduced. We also reported sequential morphological changes and loss of viability upon plasma treatment. However, the studies were carried out in batch cultures. The use of a continuous culture results in a more homogenous environment ensuring reproducible biofilm growth. The aim of this work was to study plasma-mediated inactivation of P. aeruginosa biofilms grown on borosilicate in a continuous culture system. In this paper we show that biofilms grown on glass under continuous culture can be inactivated by using gas discharge plasma. Both biofilm architecture and cell culturability are impacted by the plasma treatment. The inactivation kinetics is similar to previously described ones and cells go through sequential changes ranging from minimal modification without loss of viability at short plasma exposure times, to major structure and viability loss at longer exposure times. We report that changes in biofilm structure leading to the loss of culturability and viability are related to a decrease of the biofilm matrix adhesiveness. To our knowledge, there has been no attempt to evaluate the inactivation

  11. Plasma-mediated inactivation of Pseudomonas aeruginosa biofilms grown on borosilicate surfaces under continuous culture system.

    Directory of Open Access Journals (Sweden)

    Kurt G Vandervoort

    Full Text Available Biofilms are microbial communities attached to a surface and embedded in a matrix composed of exopolysaccharides and excreted nucleic acids. Bacterial biofilms are responsible for undesirable effects such as disease, prostheses colonization, biofouling, equipment damage, and pipe plugging. Biofilms are also more resilient than free-living cells to regular sterilization methods and therefore it is indispensable to develop better ways to control and remove them. The use of gas discharge plasmas is a good alternative since plasmas contain a mixture of reactive agents well-known for their decontamination potential against free microorganisms. We have previously reported that Pseudomonas aeruginosa biofilms were inactivated after a 1-min plasma exposure. We determined that the adhesiveness and the thickness of Pseudomonas biofilms grown on borosilicate were reduced. We also reported sequential morphological changes and loss of viability upon plasma treatment. However, the studies were carried out in batch cultures. The use of a continuous culture results in a more homogenous environment ensuring reproducible biofilm growth. The aim of this work was to study plasma-mediated inactivation of P. aeruginosa biofilms grown on borosilicate in a continuous culture system. In this paper we show that biofilms grown on glass under continuous culture can be inactivated by using gas discharge plasma. Both biofilm architecture and cell culturability are impacted by the plasma treatment. The inactivation kinetics is similar to previously described ones and cells go through sequential changes ranging from minimal modification without loss of viability at short plasma exposure times, to major structure and viability loss at longer exposure times. We report that changes in biofilm structure leading to the loss of culturability and viability are related to a decrease of the biofilm matrix adhesiveness. To our knowledge, there has been no attempt to evaluate the

  12. Biological processes for environmental control of effluent streams in the nuclear fuel cycle. [Denitrification; removal of heavy metals

    Energy Technology Data Exchange (ETDEWEB)

    Shumate, II, S E; Hancher, C W; Strandberg, G W; Scott, C D

    1978-01-01

    Nitrates and radioactive heavy metals need to be removed from aqueous effluent streams in the fuel cycle. Biological methods are being developed for reducing nitrate or nitrite to N/sub 2/ gas and for decreasing dissolved metal concentration to less than 1 g/m/sup 3/. Fluidized-bed denitrification bioreactors are being tested. Removal of uranium from solution by Saccharomyces cerevisiae and Pseudomonas aeruginosa was studied. (DLC)

  13. Toxicogenomic response of Pseudomonas aeruginosa to ortho-phenylphenol

    Directory of Open Access Journals (Sweden)

    Toghrol Freshteh

    2008-10-01

    Full Text Available Abstract Background Pseudomonas aeruginosa (P. aeruginosa is the most common opportunistic pathogen implicated in nosocomial infections and in chronic lung infections in cystic fibrosis patients. Ortho-phenylphenol (OPP is an antimicrobial agent used as an active ingredient in several EPA registered disinfectants. Despite its widespread use, there is a paucity of information on its target molecular pathways and the cellular responses that it elucidates in bacteria in general and in P. aeruginosa in particular. An understanding of the OPP-driven gene regulation and cellular response it elicits will facilitate more effective utilization of this antimicrobial and possibly lead to the development of more effective disinfectant treatments. Results Herein, we performed a genome-wide transcriptome analysis of the cellular responses of P. aeruginosa exposed to 0.82 mM OPP for 20 and 60 minutes. Our data indicated that OPP upregulated the transcription of genes encoding ribosomal, virulence and membrane transport proteins after both treatment times. After 20 minutes of exposure to 0.82 mM OPP, genes involved in the exhibition of swarming motility and anaerobic respiration were upregulated. After 60 minutes of OPP treatment, the transcription of genes involved in amino acid and lipopolysaccharide biosynthesis were upregulated. Further, the transcription of the ribosome modulation factor (rmf and an alternative sigma factor (rpoS of RNA polymerase were downregulated after both treatment times. Conclusion Results from this study indicate that after 20 minutes of exposure to OPP, genes that have been linked to the exhibition of anaerobic respiration and swarming motility were upregulated. This study also suggests that the downregulation of the rmf and rpoS genes may be indicative of the mechanism by which OPP causes decreases in cell viability in P. aeruginosa. Consequently, a protective response involving the upregulation of translation leading to the

  14. High-resolution time series of Pseudomonas aeruginosa gene expression and rhamnolipid secretion through growth curve synchronization

    Directory of Open Access Journals (Sweden)

    Xavier João B

    2011-06-01

    Full Text Available Abstract Background Online spectrophotometric measurements allow monitoring dynamic biological processes with high-time resolution. Contrastingly, numerous other methods require laborious treatment of samples and can only be carried out offline. Integrating both types of measurement would allow analyzing biological processes more comprehensively. A typical example of this problem is acquiring quantitative data on rhamnolipid secretion by the opportunistic pathogen Pseudomonas aeruginosa. P. aeruginosa cell growth can be measured by optical density (OD600 and gene expression can be measured using reporter fusions with a fluorescent protein, allowing high time resolution monitoring. However, measuring the secreted rhamnolipid biosurfactants requires laborious sample processing, which makes this an offline measurement. Results Here, we propose a method to integrate growth curve data with endpoint measurements of secreted metabolites that is inspired by a model of exponential cell growth. If serial diluting an inoculum gives reproducible time series shifted in time, then time series of endpoint measurements can be reconstructed using calculated time shifts between dilutions. We illustrate the method using measured rhamnolipid secretion by P. aeruginosa as endpoint measurements and we integrate these measurements with high-resolution growth curves measured by OD600 and expression of rhamnolipid synthesis genes monitored using a reporter fusion. Two-fold serial dilution allowed integrating rhamnolipid measurements at a ~0.4 h-1 frequency with high-time resolved data measured at a 6 h-1 frequency. We show how this simple method can be used in combination with mutants lacking specific genes in the rhamnolipid synthesis or quorum sensing regulation to acquire rich dynamic data on P. aeruginosa virulence regulation. Additionally, the linear relation between the ratio of inocula and the time-shift between curves produces high-precision measurements of

  15. NET formation induced by Pseudomonas aeruginosa cystic fibrosis isolates measured as release of myeloperoxidase-DNA and neutrophil elastase-DNA complexes.

    Science.gov (United States)

    Yoo, Dae-goon; Floyd, Madison; Winn, Matthew; Moskowitz, Samuel M; Rada, Balázs

    2014-08-01

    Cystic fibrosis (CF) airway disease is characterized by Pseudomonas aeruginosa infection and recruitment of neutrophil granulocytes. Neutrophil granule components (myeloperoxidase (MPO), human neutrophil elastase (HNE)), extracellular DNA and P. aeruginosa can all be found in the CF respiratory tract and have all been associated with worsening CF lung function. Pseudomonas-induced formation of neutrophil extracellular traps (NETs) offers a likely mechanism for release of MPO, HNE and DNA from neutrophils. NETs are composed of a DNA backbone decorated with granule proteins like MPO and HNE. Here we sought to examine whether CF clinical isolates of Pseudomonas are capable of inducing NET release from human neutrophil granulocytes. We used two methods to quantify NETs. We modified a previously employed ELISA that detects MPO-DNA complexes and established a new HNE-DNA ELISA. We show that these methods reliably quantify MPO-DNA and HNE-DNA complexes, measures of NET formation. We have found that CF isolates of P. aeruginosa stimulate robust respiratory burst and NET release in human neutrophils. By comparing paired "early" and "late" bacterial isolates obtained from the same CF patient we have found that early isolates induced significantly more NET release than late isolates. Our data support that Pseudomonas-induced NET release represents an important mechanism for release of neutrophil-derived CF inflammatory mediators, and confirm that decreased induction of NET formation is required for long-term adaptation of P. aeruginosa to CF airways.

  16. Removal of root filling materials.

    LENUS (Irish Health Repository)

    Duncan, H.F. Chong, B.S.

    2011-05-01

    Safe, successful and effective removal of root filling materials is an integral component of non-surgical root canal re-treatment. Access to the root canal system must be achieved in order to negotiate to the canal terminus so that deficiencies in the original treatment can be rectified. Since a range of materials have been advocated for filling root canals, different techniques are required for their removal. The management of commonly encountered root filling materials during non-surgical re-treatment, including the clinical procedures necessary for removal and the associated risks, are reviewed. As gutta-percha is the most widely used and accepted root filling material, there is a greater emphasis on its removal in this review.

  17. Parathyroid gland removal

    Science.gov (United States)

    Removal of parathyroid gland; Parathyroidectomy; Hyperparathyroidism - parathyroidectomy; PTH - parathyroidectomy ... and pain-free) for this surgery. Usually the parathyroid glands are removed using a 2- to 4- ...

  18. Pseudomonas aeruginosa host-adaptation in cystic fibrosis patients

    DEFF Research Database (Denmark)

    Rau, Martin Holm

    Pseudomonas aeruginosa is an opportunistic pathogen capable of transition from an environmental lifestyle to a host-associated lifestyle, as exemplified in the life-long airway infection of cystic fibrosis (CF) patients. Long-term infection is associated with extensive genetic adaptation of P....... aeruginosa towards the CF airway environment generating variants with markedly altered phenotypes. Gaining insight into this adaptation process has great clinical relevance but simultaneously has the potential to increase our understanding of bacterial adaptation to a host environment. This has been...... to unravel the early adaptive processes possibly securing bacterial persistence. In this early stage, clinical isolates displayed few adaptive events however these included phenotypes often observed in late chronic infection isolates including the conversion to a mucoid phenotype and increased antibiotic...

  19. Production and characterization of the slime polysaccharide of Pseudomonas aeruginosa.

    Science.gov (United States)

    Evans, L R; Linker, A

    1973-11-01

    The slime polysaccharides produced by Pseudomonas aeruginosa isolated from a variety of human infections were investigated. Slime production in culture seemed optimal when adequate amounts of carbohydrate were present and under conditions of either high osmotic pressure or inadequate protein supply. The polysaccharides produced by the organisms were similar to each other, to the slime of Azotobacter vinelandii, and to seaweed alginic acids. They were composed of beta-1,4-linked d-mannuronic acid residues and variable amounts of its 5-epimer l-guluronic acid. All bacterial polymers contained o-acetyl groups which are absent in the alginates. The polysaccharides differed considerably in the ratio of mannuronic to guluronic acid content and in the number of o-acetyl groups. The particular composition of the slime was not found to be characteristic for the disease process from which the mucoid variants of P. aeruginosa were obtained.

  20. Microbial Transformation of Clarias gariepinus Oil by Psuedomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Muhammad Nor Omar

    2014-09-01

    Full Text Available Bio transformation of fatty acid from Malaysian catfish, Clarias gariepinus oil was carried out using Pseudomonas aeruginosa. The lipid from freeze-dried catfish flesh was extracted using a modified Folch method with chloroform-methanol mixture as an extracting solvent. The crude lipid substrate was added to P. aeruginosa culture and incubated for 4 days. After conversion, the products were analyzed by using GC-MS instrument. The result showed that 7,10-dihydroxy-8(E-octadecenoic acid (DHOD were abundantly found in the product. The hydroxyl derivative increased while fatty acid contents decreased after bio transformation process. It can be concluded that the bacterial cells had transformed the fatty acids to yield hydroxyl metabolite which can be utilized as

  1. Flagellation of Pseudomonas aeruginosa in newly divided cells

    Science.gov (United States)

    Zhao, Kun; Lee, Calvin; Anda, Jaime; Wong, Gerard

    2015-03-01

    For monotrichous bacteria, Pseudomonas aeruginosa, after cell division, one daughter cell inherits the old flagellum from its mother cell, and the other grows a new flagellum during or after cell division. It had been shown that the new flagellum grows at the distal pole of the dividing cell when the two daughter cells haven't completely separated. However, for those daughter cells who grow new flagella after division, it still remains unknown at which pole the new flagellum will grow. Here, by combining our newly developed bacteria family tree tracking techniques with genetic manipulation method, we showed that for the daughter cell who did not inherit the old flagellum, a new flagellum has about 90% chances to grow at the newly formed pole. We proposed a model for flagellation of P. aeruginosa.

  2. Pseudomonas aeruginosa dose response and bathing water infection.

    Science.gov (United States)

    Roser, D J; van den Akker, B; Boase, S; Haas, C N; Ashbolt, N J; Rice, S A

    2014-03-01

    Pseudomonas aeruginosa is the opportunistic pathogen mostly implicated in folliculitis and acute otitis externa in pools and hot tubs. Nevertheless, infection risks remain poorly quantified. This paper reviews disease aetiologies and bacterial skin colonization science to advance dose-response theory development. Three model forms are identified for predicting disease likelihood from pathogen density. Two are based on Furumoto & Mickey's exponential 'single-hit' model and predict infection likelihood and severity (lesions/m2), respectively. 'Third-generation', mechanistic, dose-response algorithm development is additionally scoped. The proposed formulation integrates dispersion, epidermal interaction, and follicle invasion. The review also details uncertainties needing consideration which pertain to water quality, outbreaks, exposure time, infection sites, biofilms, cerumen, environmental factors (e.g. skin saturation, hydrodynamics), and whether P. aeruginosa is endogenous or exogenous. The review's findings are used to propose a conceptual infection model and identify research priorities including pool dose-response modelling, epidermis ecology and infection likelihood-based hygiene management.

  3. The implication of Pseudomonas aeruginosa biofilms in infections

    DEFF Research Database (Denmark)

    Rybtke, Morten T; Jensen, Peter Østrup; Høiby, Niels

    2011-01-01

    of infection in the lungs of cystic fibrosis patients and in chronic wounds. In this review we address the molecular basis of biofilm development by P. aeruginosa as well as the mechanisms employed by this bacterium in the increased tolerance displayed against antimicrobials. The complex build......-up of the extracellular matrix encasing the biofilm-associated bacteria as well as the elaborate signaling mechanisms employed by the bacterium enables it to withstand the continuous stresses imposed by the immune defense and administered antibiotics resulting in a state of chronic inflammation that damages the host....... The immune response leading to this chronic inflammation is described. Finally, novel treatment strategies against P. aeruginosa are described including, quorum-sensing inhibition and induced biofilm-dispersion. The tolerance towards currently available antimicrobials calls for development of alternative...

  4. [Water used for hemodialysis equipment: where is Pseudomonas aeruginosa?].

    Science.gov (United States)

    Ducki, Sébastien; Francini, Nicolas; Blech, Marie-Françoise

    2005-05-01

    The water used in dilution of the dialysis solutions constitutes an essential element of the efficiency and the safety of this therapeutics. Water must be specifically treated, and some technical rules must be respected, such as disinfection of the equipment for water treatment, to guarantee a satisfying level for whole the installation. This article reports the investigations, which were led to find the spring of Pseudomonas aeruginosa which contamined in a recurring way the water feeding dialysis equipment. The observation of samples'chronology and an analysis of the sanitary pad suggested a contamination during disinfection. Sample of residual water from the pump used for the injection of Dialox identified this reservoir as origin of the contamination. To stop this contamination by P. aeruginosa, a pump maintenance revision and purges of the system were used.

  5. An update on Pseudomonas aeruginosa biofilm formation, tolerance, and dispersal

    DEFF Research Database (Denmark)

    Harmsen, Morten; Yang, Liang; Pamp, Sünje Johanna

    2010-01-01

    . aeruginosa biofilms. The second messenger, c-di-GMP, is established as an important regulator of the synthesis of polysaccharide and protein components of the biofilm matrix. Extracellular DNA is shown to be an essential component of the biofilm matrix. It has become apparent that biofilm formation involves......We review the recent advances in the understanding of the Pseudomonas aeruginosa biofilm lifestyle from studies using in vitro laboratory setups such as flow chambers and microtiter trays. Recent work sheds light on the role of nutrients, motility, and quorum sensing in structure formation in P...... interactions between different subpopulations. The molecular mechanisms underlying the tolerance of biofilm bacteria to antimicrobial agents are beginning to be unraveled, and new knowledge has been obtained regarding the environmental cues and regulatory mechanisms involved in biofilm dispersal....

  6. High Diversity and Novel Species of Pseudomonas aeruginosa Bacteriophages

    OpenAIRE

    Sepúlveda-Robles, Omar; Kameyama, Luis; Guarneros, Gabriel

    2012-01-01

    The diversity of Pseudomonas aeruginosa bacteriophages was investigated using a collection of 68 phages isolated from Central Mexico. Most of the phages carried double-stranded DNA (dsDNA) genomes and were classified into 12 species. Comparison of the genomes of selected archetypal phages with extant sequences in GenBank resulted in the identification of six novel species. This finding increased the group diversity by ∼30%. The great diversity of phage species could be related to the ubiquito...

  7. The action of Pseudomonas aeruginosa biofilms in intrinsic drug resistance

    Institute of Scientific and Technical Information of China (English)

    XIE Yi; JIA Wen-xiang; ZENG Wei; YANG Wei-qing; CHENG Xi; LI Xue-ru; WANG Lan-lan; KANG Mei; ZHANG Zai-rong

    2005-01-01

    Background There is a growing interest in studying the relationship between intrinsic resistance and biofilms resistance to drugs. However, the relationship still remains unclear in the macroscopic bacterial growth. Our study is to illuminate the change of bacterial drug resistance of gyrA mutant and active efflux pump during the development of Pseudomonas aeruginosa (P. aeruginosa) biofilms. Methods The strains of type Ⅱ topoisomerase gene mutant (gyrA mutant) and multidrug resistance (MDR) efflux pump were clinical isolates and detected by polymerase chain reaction (PCR). The process of bacterial biofilms development was observed by scanning electron microscope. Triparental mating experiments were performed to transfer report gene of green fluorescent protein (GFP) into P. aeruginosa biofilms strains and followed by analysis of bacterial survival rate between intrinsic resistance and biofilms resistance.Results The fluorescent strains with pGFPuv could develop mature biofilms on Teflon surface. Before a period of 72 hours, the survival rate of biofilms bacteria and intrinsic resistance strains in ciprofloxacin solution was significantly different (P0.05). The carbonyl cyanide m-chlorophenylhydrazone and azithromycin could significantly reduce the drug resistance of biofilm strains and efflux pump strains.Conclusions In the development of P. aeruginosa biofilms, the strains of gyrA mutation and MDR efflux could be conferred with new level of drug resistance. When co-cultured mutated strains with biofilm strains, biofilms may play a major role in bacterial resistance. But after 72 hours incubation (a mature biofilms had been developed), there was no clearly difference between the number of mutant strains and biofilm strains.

  8. Functionalized polyanilines disrupt Pseudomonas aeruginosa and Staphylococcus aureus biofilms.

    Science.gov (United States)

    Gizdavic-Nikolaidis, Marija R; Pagnon, Joanne C; Ali, Naseem; Sum, Reuben; Davies, Noel; Roddam, Louise F; Ambrose, Mark

    2015-12-01

    The purpose of the present study was to investigate the antimicrobial effects of functionalized polyanilines (fPANIs) against stationary phase cells and biofilms of Pseudomonas aeruginosa and Staphylococcus aureus using homopolymer of sulfanilic acid (poly-SO3H) as a model. The chemically synthesized poly-SO3H was characterized using Fourier Transform Infra-Red (FTIR) and Ultraviolet-Visible (UV-Vis) spectroscopies. The molecular weight (Mw) and elemental analysis of homopolymer poly-SO3H were also examined. We found that poly-SO3H was bactericidal against stationary phase cells of P. aeruginosa and S. aureus at a concentration of 20 mgml(-1). Surprisingly, we discovered that the same concentration (20 mgml(-1)) of poly-SO3H significantly disrupted and killed bacterial cells present in pre-established forty-eight hour static biofilms of these organisms, as shown by crystal violet and bacterial live/dead fluorescence staining assays. In support of these data, poly-SO3H extensively diminished the expression of bacterial genes related to biofilm formation in stationary phase cells of P. aeruginosa, and seemed to greatly reduce the amount of the quorum sensing molecule N-(3-oxododecanoyl)-l-homoserine lactone (3OC12-HSL) able to be recovered from biofilms of this organism. Furthermore, we found that poly-SO3H was able to effectively penetrate and kill cells in biofilms formed by the P. aeruginosa (AESIII) isolate derived from the sputum of a cystic fibrosis patient. Taken together, the results of the present study emphasise the broad antimicrobial activities of fPANI, and suggest that they could be developed further and used in some novel ways to construct medical devices and/or industrial equipment that are refractory to colonization by biofilm-forming bacteria.

  9. Production and characterization of rhamnolipids from Pseudomonas aeruginosa san ai

    OpenAIRE

    Rikalovic Milena G.; Gojgic-Cvijovic Gordana; Vrvic Miroslav M.; Karadzic Ivanka

    2012-01-01

    Production and characterization of rhamnolipid biosurfactant obtained by strain Pseudomonas aeruginosa san ai was investigated. With regard to carbon and nitrogen source several media were tested to enhance production of rhamnolipids. Phosphate-limited proteose peptone-ammonium salt (PPAS) medium supplemented with sun flower oil as a source of carbon and mineral ammonium chloride and peptone as a nitrogen source greatly improved rhamnolipid production, from 0.15 on basic PPAS (C/N ratio...

  10. Phage selection restores antibiotic sensitivity in MDR Pseudomonas aeruginosa

    OpenAIRE

    Chan, Benjamin K.; Mark Sistrom; Wertz, John E.; Kaitlyn E. Kortright; Deepak Narayan; Turner, Paul E.

    2016-01-01

    Increasing prevalence and severity of multi-drug-resistant (MDR) bacterial infections has necessitated novel antibacterial strategies. Ideally, new approaches would target bacterial pathogens while exerting selection for reduced pathogenesis when these bacteria inevitably evolve resistance to therapeutic intervention. As an example of such a management strategy, we isolated a lytic bacteriophage, OMKO1, (family Myoviridae) of Pseudomonas aeruginosa that utilizes the outer membrane porin M (Op...

  11. Emergence of colistin resistant Pseudomonas aeruginosa at Tabriz hospitals, Iran

    Directory of Open Access Journals (Sweden)

    Hamid Reza Goli

    2016-03-01

    Full Text Available Background and Objectives: The prevalence of multidrug resistant Pseudomonas aeruginosa is the main reason of new drugs resurgence such as colistin. The main objectives of this study were to determine the antibiotic resistance pattern and the rate of colistin resistance along with its correlation with overexpression of MexAB-OprM and MexXY-OprM efflux pumps among P. aeruginosa isolates.Materials and Methods: Hundred clinical isolates were collected from 100 patients during 6 months in 2014. Susceptibility to the eight antibiotics was investigated using Kirby-Bauer and agar dilution methods. The Quantitative Real-time PCR was used to determine the expression levels of efflux genes.Results: Resistance rates to various antibiotics were as follows: ticarcillin (73%, ciprofloxacin (65%, aztreonam (60%, ceftazidime (55%, gentamicin (55%, imipenem (49%, piperacillin/tazobactam (34% and colistin (2%. In disk diffusion method, only two isolates were non susceptible to colistin, however in agar dilution method the two isolates were confirmed as resistant and two others were intermediate resistant. Sixty eight (68% isolates were multi-drug resistant and 10 isolates were susceptible to all tested antibiotics. Both colistin resistant isolates showed overexpression of both efflux pumps, but two intermediate resistant isolates exhibited reduction of efflux genes expression.Conclusions: Emergence of colistin resistance is increasing in P. aeruginosa indicating great challenge in the treatment of infections caused by MDR strains of this organism in Iran. ParRS may promote either induced or constitutive resistance to colistin through the activation of distinct mechanisms such as MDR efflux pumps, and LPS modification. Keywords: Pseudomonas aeruginosa, Multi drug resistant, Colistin, MexAB-OprM, MexXY-OprM

  12. AMINOGLYCOSIDE RESISTANCE GENES IN Pseudomonas aeruginosa ISOLATES FROM CUMANA, VENEZUELA

    Directory of Open Access Journals (Sweden)

    Bertinellys TEIXEIRA

    2016-01-01

    Full Text Available The enzymatic modification of aminoglycosides by aminoglycoside-acetyltransferases (AAC, aminoglycoside-adenyltransferases (AAD, and aminoglycoside-phosphotransferases (APH, is the most common resistance mechanism in P. aeruginosa and these enzymes can be coded on mobile genetic elements that contribute to their dispersion. One hundred and thirty seven P. aeruginosa isolates from the University Hospital, Cumana, Venezuela (HUAPA were evaluated. Antimicrobial susceptibility was determined by the disk diffusion method and theaac, aadB and aph genes were detected by PCR. Most of the P. aeruginosa isolates (33/137 were identified from the Intensive Care Unit (ICU, mainly from discharges (96/137. The frequency of resistant P. aeruginosaisolates was found to be higher for the aminoglycosides tobramycin and amikacin (30.7 and 29.9%, respectively. Phenotype VI, resistant to these antibiotics, was the most frequent (14/49, followed by phenotype I, resistant to all the aminoglycosides tested (12/49. The aac(6´-Ib,aphA1 and aadB genes were the most frequently detected, and the simultaneous presence of several resistance genes in the same isolate was demonstrated. Aminoglycoside resistance in isolates ofP. aeruginosa at the HUAPA is partly due to the presence of the aac(6´-Ib, aphA1 andaadB genes, but the high rates of antimicrobial resistance suggest the existence of several mechanisms acting together. This is the first report of aminoglycoside resistance genes in Venezuela and one of the few in Latin America.

  13. Pseudomonas aeruginosa extracellular products inhibit staphylococcal growth, and disrupt established biofilms produced by Staphylococcus epidermidis

    DEFF Research Database (Denmark)

    Qin, Zhiqiang; Yang, Liang; Qu, Di

    2009-01-01

    Multiple bacterial species often coexist as communities, and compete for environmental resources. Here, we describe how an opportunistic pathogen, Pseudomonas aeruginosa, uses extracellular products to interact with the nosocomial pathogen Staphylococcus epidermidis. S. epidermidis biofilms...... and planktonic cultures were challenged with P. aeruginosa supernatant cultures overnight. Results indicated that quorum-sensing-controlled factors from P. aeruginosa supernatant inhibited S. epidermidis growth in planktonic cultures. We also found that P. aeruginosa extracellular products, mainly...... in overnight cultures had no effect on established P. aeruginosa biofilms and planktonic growth. These findings reveal that P. aeruginosa extracellular products are important microbial competition factors that overcome competition with S. epidermidis, and the results may provide clues for the development...

  14. Effect of Tyrosol and Farnesol on Virulence and Antibiotic Resistance of Clinical Isolates of Pseudomonas aeruginosa.

    Science.gov (United States)

    Abdel-Rhman, Shaymaa Hassan; El-Mahdy, Areej Mostafa; El-Mowafy, Mohammed

    2015-01-01

    Mixed-species biofilms could create a protected environment that allows for survival to external antimicrobials and allows different bacterial-fungal interactions. Pseudomonas aeruginosa-Candida albicans coexistence is an example for such mixed-species community. Numerous reports demonstrated how P. aeruginosa or its metabolites could influence the growth, morphogenesis, and virulence of C. albicans. In this study, we investigated how the C. albicans quorum sensing compounds, tyrosol and farnesol, might affect Egyptian clinical isolates of P. aeruginosa regarding growth, antibiotic sensitivity, and virulence. We could demonstrate that tyrosol possesses an antibacterial activity against P. aeruginosa (10 µM inhibited more than 50% of growth after 16 h cultivation). Moreover, we could show for the first time that tyrosol strongly inhibits the production of the virulence factors hemolysin and protease in P. aeruginosa, whereas farnesol inhibits, to lower extent, hemolysin production in this bacterial pathogen. Cumulatively, tyrosol is expected to strongly affect P. aeruginosa in mixed microbial biofilm.

  15. Quorum sensing and policing of Pseudomonas aeruginosa social cheaters.

    Science.gov (United States)

    Wang, Meizhen; Schaefer, Amy L; Dandekar, Ajai A; Greenberg, E Peter

    2015-02-17

    The bacterium Pseudomonas aeruginosa is an opportunistic human pathogen that uses a quorum sensing signal cascade to activate expression of dozens of genes when sufficient population densities have been reached. Quorum sensing controls production of several key virulence factors, including secreted proteases such as elastase. Cooperating groups of bacteria growing on protein are susceptible to social cheating by quorum-sensing defective mutants. A possible way to restrict cheater emergence is by policing where cooperators produce costly goods to sanction or punish cheats. The P. aeruginosa LasR-LasI quorum sensing system controls genes including those encoding proteases and also those encoding a second quorum-sensing system, the RhlR-RhlI system, which controls numerous genes including those for cyanide production. By using RhlR quorum sensing mutants and cyanide synthesis mutants, we show that cyanide production is costly and cyanide-producing cooperators use cyanide to punish LasR-null social cheaters. Cooperators are less susceptible to cyanide than are LasR mutants. These experiments demonstrate policing in P. aeruginosa, provide a mechanistic understanding of policing, and show policing involves the cascade organization of the two quorum sensing systems in this bacterium.

  16. Proteolytic regulation of alginate overproduction in Pseudomonas aeruginosa.

    Science.gov (United States)

    Damron, F Heath; Goldberg, Joanna B

    2012-05-01

    Pseudomonas aeruginosa, a Gram-negative bacterium, is a significant opportunistic pathogen associated with skin and soft tissue infections, nosocomial pneumonia and sepsis. In addition, it can chronically colonize the lungs of cystic fibrosis (CF) patients. Overproduction of the exopolysaccharide called alginate provides P. aeruginosa with a selective advantage and facilitates survival in the CF lung. The in vitro phenotype of alginate overproduction observed on solid culture media is referred to as mucoid. Expression of the alginate machinery and biosynthetic enzymes are controlled by the extracytoplasmic sigma factor, σ(22) (AlgU/T). The key negative regulator of both σ(22) activity and the mucoid phenotype is the cognate anti-sigma factor MucA. MucA sequesters σ(22) to the inner membrane inhibiting the sigma factor's transcriptional activity. The well-studied mechanism for transition to the mucoid phenotype is mutation of mucA, leading to loss of MucA function and therefore activation of σ(22) . Recently, regulated intramembrane proteolysis (RIP) has been recognized as a mechanism whereby proteolysis of the anti-sigma factor MucA leads to active σ(22) allowing P. aeruginosa to respond to environmental stress conditions by overproduction of alginate. The goal of this review is to illuminate the pathways leading to RIP that have been identified and proposed.

  17. Mechanism of azithromycin inhibition of HSL synthesis in Pseudomonas aeruginosa.

    Science.gov (United States)

    Zeng, Jianming; Zhang, Ni; Huang, Bin; Cai, Renxin; Wu, Binning; E, Shunmei; Fang, Chengcai; Chen, Cha

    2016-04-14

    Pseudomonas aeruginosa is an opportunistic pathogen and a leading cause of nosocomial infections. Unfortunately, P. aeruginosa has low antibiotic susceptibility due to several chromosomally encoded antibiotic resistance genes. Hence, we carried out mechanistic studies to determine how azithromycin affects quorum sensing and virulence in P. aeruginosa. lasI and rhlI single and double mutants were constructed. We then undertook a quantitative approach to determine the optimal concentration of azithromycin and culture time that can affect the expression of HSLs. Furthermore, based on the above results, the effect on quorum sensing was analyzed at a transcriptional level. It was found that 2 μg/mL azithromycin caused a 79% decrease in 3-oxo-C12-HSL secretion during cultivation, while C4-HSL secretion was strongly repressed in the early stages. Azithromycin acts on ribosomes; to determine whether this can elicit alternative modes of gene expression, transcriptional regulation of representative virulence genes was analyzed. We propose a new relationship for lasI and rhlI: lasI acts as a cell density sensor, and rhlI functions as a fine-tuning mechanism for coordination between different quorum sensing systems.

  18. Functional study of elafin cleaved by Pseudomonas aeruginosa metalloproteinases.

    LENUS (Irish Health Repository)

    Guyot, Nicolas

    2010-06-01

    Elafin is a 6-kDa innate immune protein present at several epithelial surfaces including the pulmonary epithelium. It is a canonical protease inhibitor of two neutrophil serine proteases [neutrophil elastase (NE) and proteinase 3] with the capacity to covalently bind extracellular matrix proteins by transglutamination. In addition to these properties, elafin also possesses antimicrobial and immunomodulatory activities. The aim of the present study was to investigate the effect of Pseudomonas aeruginosa proteases on elafin function. We found that P. aeruginosa PAO1-conditioned medium and two purified Pseudomonas metalloproteases, pseudolysin (elastase) and aeruginolysin (alkaline protease), are able to cleave recombinant elafin. Pseudolysin was shown to inactivate the anti-NE activity of elafin by cleaving its protease-binding loop. Interestingly, antibacterial properties of elafin against PAO1 were found to be unaffected after pseudolysin treatment. In contrast to pseudolysin, aeruginolysin failed to inactivate the inhibitory properties of elafin against NE. Aeruginolysin cleaves elafin at the amino-terminal Lys6-Gly7 peptide bond, resulting in a decreased ability to covalently bind purified fibronectin following transglutaminase activity. In conclusion, this study provides evidence that elafin is susceptible to proteolytic cleavage at alternative sites by P. aeruginosa metalloproteinases, which can affect different biological functions of elafin.

  19. Strategies for improved rhamnolipid production by Pseudomonas aeruginosa PA1.

    Science.gov (United States)

    Soares Dos Santos, Alexandre; Pereira, Nei; Freire, Denise M G

    2016-01-01

    Rhamnolipids are biosurfactants with potential for diversified industrial and environmental uses. The present study evaluated three strategies for increasing the production of rhamnolipid-type biosurfactants produced by Pseudomonas aeruginosa strain PA1. The influence of pH, the addition of P. aeruginosa spent culture medium and the use of a fed-batch process were examined. The culture medium adjusted to pH 7.0 was the most productive. Furthermore, the pH of the culture medium had a measurable effect on the ratio of synthesized mono- and dirhamnolipids. At pH values below 7.3, the proportion of monorhamnolipids decreased from 45 to 24%. The recycling of 20% of the spent culture medium in where P. aeruginosa was grown up to the later stationary phase was responsible for a 100% increase in rhamnolipid volumetric productivity in the new culture medium. Finally, the use of fed-batch operation under conditions of limited nitrogen resulted in a 3.8-fold increase in the amount of rhamnolipids produced (2.9 g L(-1)-10.9 g L(-1)). These results offer promising pathways for the optimization of processes for the production of rhamnolipids.

  20. Evolutionary genomics of epidemic and nonepidemic strains of Pseudomonas aeruginosa.

    Science.gov (United States)

    Dettman, Jeremy R; Rodrigue, Nicolas; Aaron, Shawn D; Kassen, Rees

    2013-12-24

    Pseudomonas aeruginosa is an opportunistic pathogen of humans and is a major cause of morbidity and mortality in patients with cystic fibrosis (CF). Prolonged infection of the respiratory tract can lead to adaptation of the pathogen to the CF lung environment. To examine the general patterns of adaptation associated with chronic infection, we obtained genome sequences from a collection of P. aeruginosa isolated from airways of patients with CF. Our analyses support a nonclonal epidemic population structure, with a background of unique, recombining genotypes, and the rare occurrence of successful epidemic clones. We present unique genome sequence evidence for the intercontinental spread of an epidemic strain shared between CF clinics in the United Kingdom and North America. Analyses of core and accessory genomes identified candidate genes and important functional pathways associated with adaptive evolution. Many genes of interest were involved in biological functions with obvious roles in this pathosystem, such as biofilm formation, antibiotic metabolism, pathogenesis, transport, reduction/oxidation, and secretion. Key factors driving the adaptive evolution of this pathogen within the host appear to be the presence of oxidative stressors and antibiotics. Regions of the accessory genome unique to the epidemic strain were enriched for genes in transporter families that efflux heavy metals and antibiotics. The epidemic strain was significantly more resistant than nonepidemic strains to three different antibiotics. Multiple lines of evidence suggest that selection imposed by the CF lung environment has a major influence on genomic evolution and the genetic characteristics of P. aeruginosa isolates causing contemporary infection.

  1. Arsenic efflux from Microcystis aeruginosa under different phosphate regimes.

    Directory of Open Access Journals (Sweden)

    Changzhou Yan

    Full Text Available Phytoplankton plays an important role in arsenic speciation, distribution, and cycling in freshwater environments. Little information, however, is available on arsenic efflux from the cyanobacteria Microcystis aeruginosa under different phosphate regimes. This study investigated M. aeruginosa arsenic efflux and speciation by pre-exposing it to 10 µM arsenate or arsenite for 24 h during limited (12 h and extended (13 d depuration periods under phosphate enriched (+P and phosphate depleted (-P treatments. Arsenate was the predominant species detected in algal cells throughout the depuration period while arsenite only accounted for no greater than 45% of intracellular arsenic. During the limited depuration period, arsenic efflux occurred rapidly and only arsenate was detected in solutions. During the extended depuration period, however, arsenate and dimethylarsinic acid (DMA were found to be the two predominant arsenic species detected in solutions under -P treatments, but arsenate was the only species detected under +P treatments. Experimental results also suggest that phosphorus has a significant effect in accelerating arsenic efflux and promoting arsenite bio-oxidation in M. aeruginosa. Furthermore, phosphorus depletion can reduce arsenic efflux from algal cells as well as accelerate arsenic reduction and methylation. These findings can contribute to our understanding of arsenic biogeochemistry in aquatic environments and its potential environmental risks under different phosphorus levels.

  2. Experimental study on Cr(Ⅵ) reduction by Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    LIU Yun-guo; XU Wei-hua; ZENG Guang-ming; TANG Chun-fang; LI Cheng-feng

    2004-01-01

    Investigation on Cr(Ⅵ) reduction was conducted using Pseudomonas aeruginosa. The study demonstrated that the Cr(Ⅵ) can be effectively reduced to Cr(Ⅲ) by Pseudomonas aeruginosa. The effects of the factors affecting Cr(Ⅵ) reduction rate including carbon source type, pH, initial Cr(Ⅵ) concentration and amount of cells inoculum were thoroughly studied. Malate was found to yield maximum biotransformation, followed by succinate and glucose, with the reduction rate of 60.86%, 43.76% and 28.86% respectively. The optimum pH for Cr(Ⅵ) reduction was 7.0, with reduction efficiency of 61.71% being achieved. With the increase of initial Cr(Ⅵ) concentration, the rate of Cr(Ⅵ) reduction decreased. The reduction was inhibited strongly when the initial Cr(Ⅵ) concentration increased to 157 mg/L. As the amount of cells inoculum increased, the rate of Cr(Ⅵ) reduction also increased. The mechanism of Cr(Ⅵ) reduction and final products were also analysed. The results suggested that the soluble enzymes appear to be responsible for Cr(Ⅵ) reduction by Pseudomonas aeruginosa, and the reduced Cr(Ⅲ) was not precipitated in the form of Cr(OH)3.

  3. Aminoglycoside-Resistant Mutation of Pseudomonas aeruginosa Defective in Cytochrome c552 and Nitrate Reductase

    OpenAIRE

    Bryan, L E; Nicas, Thalia; Holloway, B W; Crowther, Carol

    1980-01-01

    A gentamicin-resistant mutant of Pseudomonas aeruginosa PAO503 was selected after ethyl methane sulfonate mutagenesis. The strain, P. aeruginosa PAO2401 had increased resistance to all aminoglycosides tested but exhibited no change for other antibiotics. The mutation designated aglA (aminoglycoside resistance) was 50% cotransducible with the 8-min ilvB,C marker on the P. aeruginosa chromosome. It showed a marked reduction in cytochrome c552 and nitrate reductase (Nar) and a change in terminal...

  4. Regulation and Function of Versatile Aerobic and Anaerobic Respiratory Metabolism in Pseudomonas aeruginosa

    OpenAIRE

    Hiroyuki eArai

    2011-01-01

    Pseudomonas aeruginosa is a ubiquitously distributed opportunistic pathogen that inhabits soil and water as well as animal-, human-, and plant-host-associated environments. The ubiquity would be attributed to its very versatile energy metabolism. P. aeruginosa has a highly branched respiratory chain terminated by multiple terminal oxidases and denitrification enzymes. Five terminal oxidases for aerobic respiration have been identified in the P. aeruginosa cells. Three of them, the cbb 3-1 oxi...

  5. Draft Genome Sequences of Pseudomonas aeruginosa Isolates from Wounded Military Personnel.

    Science.gov (United States)

    Arivett, Brock A; Ream, Dave C; Fiester, Steven E; Kidane, Destaalem; Actis, Luis A

    2016-08-11

    Pseudomonas aeruginosa, a Gram-negative bacterium that causes severe hospital-acquired infections, is grouped as an ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogen because of its extensive drug resistance phenotypes and effects on human health worldwide. Five multidrug resistant P. aeruginosa strains isolated from wounded military personnel were sequenced and annotated in this work.

  6. The Pseudomonas aeruginosa autoinducer dodecanoyl-homoserine lactone inhibits the putrescine synthesis in human cells

    DEFF Research Database (Denmark)

    Kristiansen, S.; Bjarnsholt, Thomas; Adeltoft, D.;

    2008-01-01

    . Polyamines are required for mitotic cell division and peak during this phase. The polyamine putrescine is synthesized by ornithine decarboxylase (ODC) as a rate-limiting step. The ODC enzyme concentration also peaks during the mitotic phase. This peak is mediated by translation of ODC mRNA by the ITAF45....... Finally, C-12-HSL-treated cells also had a time-course-dependent higher concentration of ODC mRNA. Based on these mitotic markers, more human cells were apparently trapped in the mitotic phase when treated with C-12-HSL. This should normally imply higher levels of putrescine. However, C-12-HSL......-treated human cells had a significantly lower concentration of putrescine and displayed a lower cell proliferation rate. In conclusion, the P. aeruginosa autoinducer C-12-oxo-HSL apparently arrests human cells in the mitotic phase by lowering the concentration of putrescine....

  7. UVC fluencies for preventative treatment of Pseudomonas aeruginosa contaminated polymer tubes

    DEFF Research Database (Denmark)

    Bak, Jimmy; Ladefoged, Søren D; Begovic, Tanja;

    2010-01-01

    .40-1.50) normally used for tubing in catheter production. Determining whether or not UVC light exposure can disinfect and maintain the intra-luminal number of colony forming units (CFUs) at an exceedingly low level and thus avoid the growth and establishment of biofilm is of interest. The use of UVC diodes......Exposing Pseudomonas aeruginosa biofilm grown on the inner surface of Teflon and silicone tubes to UVC light (265 nm) from light emitting diodes (LED) has previously been shown to substantially reduce biofilm growth. Smaller UVC fluencies were required to disinfect Teflon tubes compared to silicone...... tubes. Light propagation enhancement in tubes can be obtained if the refractive index of the intra-luminal saline solution is higher than that of the polymer. This condition is achieved by using Teflon tubes with a low refractive index (1.34) instead of the polymers with a high refractive index (1...

  8. Selection of phages and conditions for the safe phage therapy against Pseudomonas aeruginosa infections

    Institute of Scientific and Technical Information of China (English)

    Victor; Krylov; Olga; Shaburova; Elena; Pleteneva; Sergey; Krylov; Alla; Kaplan; Maria; Burkaltseva; Olga; Polygach; Elena; Chesnokova

    2015-01-01

    The emergence of multidrug-resistant bacterial pathogens forced us to consider the phage therapy as one of the possible alternative approaches to treatment. The purpose of this paper is to consider the conditions for the safe, long-term use of phage therapy against various infections caused by Pseudomonas aeruginosa. We describe the selection of the most suitable phages, their most effective combinations and some approaches for the rapid recognition of phages unsuitable for use in therapy. The benefi ts and disadvantages of the various different approaches to the preparation of phage mixtures are considered, together with the specifi c conditions that are required for the safe application of phage therapy in general hospitals and the possibilities for the development of personalized phage therapy.

  9. Selection of phages and conditions for the safe phage therapy against Pseudomonas aeruginosa infections.

    Science.gov (United States)

    Krylov, Victor; Shaburova, Olga; Pleteneva, Elena; Krylov, Sergey; Kaplan, Alla; Burkaltseva, Maria; Polygach, Olga; Chesnokova, Elena

    2015-02-01

    The emergence of multidrug-resistant bacterial pathogens forced us to consider the phage therapy as one of the possible alternative approaches to treatment. The purpose of this paper is to consider the conditions for the safe, long-term use of phage therapy against various infections caused by Pseudomonas aeruginosa. We describe the selection of the most suitable phages, their most effective combinations and some approaches for the rapid recognition of phages unsuitable for use in therapy. The benefits and disadvantages of the various different approaches to the preparation of phage mixtures are considered, together with the specific conditions that are required for the safe application of phage therapy in general hospitals and the possibilities for the development of personalized phage therapy.

  10. Pseudomonas aeruginosa multirresistente: um problema endêmico no Brasil Multidrug-resistant Pseudomonas aeruginosa: an endemic problem in Brazil

    Directory of Open Access Journals (Sweden)

    Patrícia R. Neves

    2011-08-01

    Full Text Available Relatos mundiais têm documentado a problemática da endemicidade de isolados clínicos de Pseudomonas aeruginosa multirresistente (MDR aliada a elevados índices de morbidade/mortalidade. No Brasil, surtos de infecção ocasionados por P. aeruginosa têm sido relacionados com uma disseminação clonal da espécie. Atualmente, as opções terapêuticas para o tratamento das infecções causadas por esse microrganismo são limitadas, muitas vezes restringindo-se ao uso de carbapenêmicos (p. ex., imipenem [IPM]. Assim, a resistência ao IPM é uma questão de saúde pública, uma vez que esse antibiótico é empregado como último recurso no tratamento de infecções de origem hospitalar, causadas por bactérias Gram-negativas multirresistentes. No Brasil, os principais mecanismos relacionados com fenótipos multirresistentes de P. aeruginosa são produção de metalobetalactamase (MBL do tipo SPM-1, presença de metilase 16S rRNA RmtD, perda de porina OprD e superexpressão de bombas de efluxo, o que pode explicar os altos índices de resistência a carbapenêmicos e aminoglicosídeos. A emergência de cepas com essas características é preocupante, tendo em vista a escassez de terapias efetivas no tratamento de infecções por esse patógeno. Finalmente, com base em relatos nacionais, publicados por diferentes grupos de pesquisa, podemos deduzir que a convergência de múltiplos mecanismos de resistência em P. aeruginosa tem sido um evento favorável para a seleção de diferentes clones endêmicos multirresistentes disseminados no Brasil.Global reports have documented the endemicity of multidrug-resistant (MDR Pseudomonas aeruginosa associated with high levels of morbidity/mortality. In Brazil, outbreaks of MDR P. aeruginosa have been related to clonal dissemination. Currently, therapeutic options for the treatment of these infections are restricted to carbapenemic antibiotics (i.e., imipenem [IPM]. Thus, carbapenem resistance is a public

  11. Pseudomonas aeruginosa forms Biofilms in Acute InfectionIndependent of Cell-to-Cell Signaling

    Energy Technology Data Exchange (ETDEWEB)

    Schaber, J. Andy; Triffo, W.J.; Suh, Sang J.; Oliver, Jeffrey W.; Hastert, Mary C.; Griswold, John A.; Auer, Manfred; Hamood, Abdul N.; Rumbaugh, Kendra P.

    2006-09-20

    Biofilms are bacterial communities residing within a polysaccharide matrix that are associated with persistence and antibiotic resistance in chronic infections. We show that the opportunistic pathogen Pseudomonas aeruginosa forms biofilms within 8 hours of infection in thermally-injured mice, demonstrating that biofilms contribute to bacterial colonization in acute infections. P. aeruginosa biofilms were visualized within burned tissue surrounding blood vessels and adipose cells. Although quorum sensing (QS), a bacterial signaling mechanism, coordinates differentiation of biofilms in vitro, wild type and QS-deficient P. aeruginosa formed similar biofilms in vivo. Our findings demonstrate that P. aeruginosa forms biofilms on specific host tissues independent of QS.

  12. Continued transmission of Pseudomonas aeruginosa from a wash hand basin tap in a critical care unit.

    Science.gov (United States)

    Garvey, M I; Bradley, C W; Tracey, J; Oppenheim, B

    2016-09-01

    Pseudomonas aeruginosa is an important nosocomial pathogen, colonizing hospital water supplies including taps and sinks. We report a cluster of P. aeruginosa acquisitions during a period of five months from tap water to patients occupying the same burns single room in a critical care unit. Pseudomonas aeruginosa cultured from clinical isolates from four different patients was indistinguishable from water strains by pulsed-field gel electrophoresis. Water outlets in critical care may be a source of P. aeruginosa despite following the national guidance, and updated guidance and improved control measures are needed to reduce the risks of transmission to patients.

  13. Pseudomonas aeruginosa Biofilm Response and Resistance to Cold Atmospheric Pressure Plasma Is Linked to the Redox-Active Molecule Phenazine.

    Science.gov (United States)

    Mai-Prochnow, Anne; Bradbury, Mark; Ostrikov, Kostya; Murphy, Anthony B

    2015-01-01

    Pseudomonas aeruginosa is an important opportunistic pathogen displaying high antibiotic resistance. Its resistance is in part due to its outstanding ability to form biofilms on a range of biotic and abiotic surfaces leading to difficult-to-treat, often long-term infections. Cold atmospheric plasma (CAP) is a new, promising antibacterial treatment to combat antibiotic-resistant bacteria. Plasma is ionized gas that has antibacterial properties through the generation of a mix of reactive oxygen and nitrogen species (RONS), excited molecules, charged particles and UV photons. Our results show the efficient removal of P. aeruginosa biofilms using a plasma jet (kINPen med), with no viable cells detected after 5 min treatment and no attached biofilm cells visible with confocal microscopy after 10 min plasma treatment. Because of its multi-factorial action, it is widely presumed that the development of bacterial resistance to plasma is unlikely. However, our results indicate that a short plasma treatment (3 min) may lead to the emergence of a small number of surviving cells exhibiting enhanced resistance to subsequent plasma exposure. Interestingly, these cells also exhibited a higher degree of resistance to hydrogen peroxide. Whole genome comparison between surviving cells and control cells revealed 10 distinct polymorphic regions, including four belonging to the redox active, antibiotic pigment phenazine. Subsequently, the interaction between phenazine production and CAP resistance was demonstrated in biofilms of transposon mutants disrupted in different phenazine pathway genes which exhibited significantly altered sensitivity to CAP.

  14. Pseudomonas aeruginosa Biofilm Response and Resistance to Cold Atmospheric Pressure Plasma Is Linked to the Redox-Active Molecule Phenazine.

    Directory of Open Access Journals (Sweden)

    Anne Mai-Prochnow

    Full Text Available Pseudomonas aeruginosa is an important opportunistic pathogen displaying high antibiotic resistance. Its resistance is in part due to its outstanding ability to form biofilms on a range of biotic and abiotic surfaces leading to difficult-to-treat, often long-term infections. Cold atmospheric plasma (CAP is a new, promising antibacterial treatment to combat antibiotic-resistant bacteria. Plasma is ionized gas that has antibacterial properties through the generation of a mix of reactive oxygen and nitrogen species (RONS, excited molecules, charged particles and UV photons. Our results show the efficient removal of P. aeruginosa biofilms using a plasma jet (kINPen med, with no viable cells detected after 5 min treatment and no attached biofilm cells visible with confocal microscopy after 10 min plasma treatment. Because of its multi-factorial action, it is widely presumed that the development of bacterial resistance to plasma is unlikely. However, our results indicate that a short plasma treatment (3 min may lead to the emergence of a small number of surviving cells exhibiting enhanced resistance to subsequent plasma exposure. Interestingly, these cells also exhibited a higher degree of resistance to hydrogen peroxide. Whole genome comparison between surviving cells and control cells revealed 10 distinct polymorphic regions, including four belonging to the redox active, antibiotic pigment phenazine. Subsequently, the interaction between phenazine production and CAP resistance was demonstrated in biofilms of transposon mutants disrupted in different phenazine pathway genes which exhibited significantly altered sensitivity to CAP.

  15. Overview of paint removal methods

    Science.gov (United States)

    Foster, Terry

    1995-04-01

    With the introduction of strict environmental regulations governing the use and disposal of methylene chloride and phenols, major components of chemical paint strippers, there have been many new environmentally safe and effective methods of paint removal developed. The new methods developed for removing coatings from aircraft and aircraft components include: mechanical methods using abrasive media such as plastic, wheat starch, walnut shells, ice and dry ice, environmentally safe chemical strippers and paint softeners, and optical methods such as lasers and flash lamps. Each method has its advantages and disadvantages, and some have unique applications. For example, mechanical and abrasive methods can damage sensitive surfaces such as composite materials and strict control of blast parameters and conditions are required. Optical methods can be slow, leaving paint residues, and chemical methods may not remove all of the coating or require special coating formulations to be effective. As an introduction to environmentally safe and effective methods of paint removal, this paper is an overview of the various methods available. The purpose of this overview is to introduce the various paint removal methods available.

  16. Induced fit on heme binding to the Pseudomonas aeruginosa cytoplasmic protein (PhuS) drives interaction with heme oxygenase (HemO)

    OpenAIRE

    2012-01-01

    Iron, an essential nutrient with limited bioavailability, requires specialized cellular mechanisms for uptake. Although iron uptake into the cytoplasm in the form of heme has been well characterized in many bacteria, the subsequent trafficking is poorly understood. The cytoplasmic heme-binding proteins belong to a structurally related family thought to have evolved as “induced fit” ligand-binding macromolecules. One member, Pseudomonas aeruginosa cytoplasmic protein (PhuS), has previously bee...

  17. Oligoribonuclease is a central feature of cyclic diguanylate signaling in Pseudomonas aeruginosa

    Science.gov (United States)

    Cohen, Dorit; Mechold, Undine; Nevenzal, Hadas; Yarmiyhu, Yafit; Randall, Trevor E.; Bay, Denice C.; Rich, Jacquelyn D.; Parsek, Matthew R.; Kaever, Volkhard; Harrison, Joe J.; Banin, Ehud

    2015-01-01

    The second messenger cyclic diguanylate (c-di-GMP) controls diverse cellular processes among bacteria. Diguanylate cyclases synthesize c-di-GMP, whereas it is degraded by c-di-GMP–specific phosphodiesterases (PDEs). Nearly 80% of these PDEs are predicted to depend on the catalytic function of glutamate-alanine-leucine (EAL) domains, which hydrolyze a single phosphodiester group in c-di-GMP to produce 5ʹ-phosphoguanylyl-(3ʹ,5ʹ)-guanosine (pGpG). However, to degrade pGpG and prevent its accumulation, bacterial cells require an additional nuclease, the identity of which remains unknown. Here we identify oligoribonuclease (Orn)—a 3ʹ→5ʹ exonuclease highly conserved among Actinobacteria, Beta-, Delta- and Gammaproteobacteria—as the primary enzyme responsible for pGpG degradation in Pseudomonas aeruginosa cells. We found that a P. aeruginosa Δorn mutant had high intracellular c-di-GMP levels, causing this strain to overexpress extracellular polymers and overproduce biofilm. Although recombinant Orn degraded small RNAs in vitro, this enzyme had a proclivity for degrading RNA oligomers comprised of two to five nucleotides (nanoRNAs), including pGpG. Corresponding with this activity, Δorn cells possessed highly elevated pGpG levels. We found that pGpG reduced the rate of c-di-GMP degradation in cell lysates and inhibited the activity of EAL-dependent PDEs (PA2133, PvrR, and purified recombinant RocR) from P. aeruginosa. This pGpG-dependent inhibition was alleviated by the addition of Orn. These data suggest that elevated levels of pGpG exert product inhibition on EAL-dependent PDEs, thereby increasing intracellular c-di-GMP in Δorn cells. Thus, we propose that Orn provides homeostatic control of intracellular pGpG under native physiological conditions and that this activity is fundamental to c-di-GMP signal transduction. PMID:26305928

  18. Expression of PPARγ and paraoxonase 2 correlated with Pseudomonas aeruginosa infection in cystic fibrosis.

    Directory of Open Access Journals (Sweden)

    Phoebe E Griffin

    Full Text Available The Pseudomonas aeruginosa quorum sensing signal molecule N-3-oxododecanoyl-l-homoserine lactone (3OC(12HSL can inhibit function of the mammalian anti-inflammatory transcription factor peroxisome proliferator activated receptor (PPARγ, and can be degraded by human paraoxonase (PON2. Because 3OC(12HSL is detected in lungs of cystic fibrosis (CF patients infected with P. aeruginosa, we investigated the relationship between P. aeruginosa infection and gene expression of PPARγ and PON2 in bronchoalveolar lavage fluid (BALF of children with CF. Total RNA was extracted from cell pellets of BALF from 43 children aged 6 months-5 years and analyzed by reverse transcription-quantitative real time PCR for gene expression of PPARγ, PON2, and P. aeruginosa lasI, the 3OC(12HSL synthase. Patients with culture-confirmed P. aeruginosa infection had significantly lower gene expression of PPARγ and PON2 than patients without P. aeruginosa infection. All samples that were culture-positive for P. aeruginosa were also positive for lasI expression. There was no significant difference in PPARγ or PON2 expression between patients without culture-detectable infection and those with non-Pseudomonal bacterial infection, so reduced expression was specifically associated with P. aeruginosa infection. Expression of both PPARγ and PON2 was inversely correlated with neutrophil counts in BALF, but showed no correlation with other variables evaluated. Thus, lower PPARγ and PON2 gene expression in the BALF of children with CF is associated specifically with P. aeruginosa infection and neutrophilia. We cannot differentiate whether this is a cause or the effect of P. aeruginosa infection, but propose that the level of expression of these genes may be a marker for susceptibility to early acquisition of P. aeruginosa in children with CF.

  19. Candida albicans Inhibits Pseudomonas aeruginosa Virulence through Suppression of Pyochelin and Pyoverdine Biosynthesis.

    Directory of Open Access Journals (Sweden)

    Eduardo Lopez-Medina

    2015-08-01

    Full Text Available Bacterial-fungal interactions have important physiologic and medical ramifications, but the mechanisms of these interactions are poorly understood. The gut is host to trillions of microorganisms, and bacterial-fungal interactions are likely to be important. Using a neutropenic mouse model of microbial gastrointestinal colonization and dissemination, we show that the fungus Candida albicans inhibits the virulence of the bacterium Pseudomonas aeruginosa by inhibiting P. aeruginosa pyochelin and pyoverdine gene expression, which plays a critical role in iron acquisition and virulence. Accordingly, deletion of both P. aeruginosa pyochelin and pyoverdine genes attenuates P. aeruginosa virulence. Heat-killed C. albicans has no effect on P. aeruginosa, whereas C. albicans secreted proteins directly suppress P. aeruginosa pyoverdine and pyochelin expression and inhibit P. aeruginosa virulence in mice. Interestingly, suppression or deletion of pyochelin and pyoverdine genes has no effect on P. aeruginosa's ability to colonize the GI tract but does decrease P. aeruginosa's cytotoxic effect on cultured colonocytes. Finally, oral iron supplementation restores P. aeruginosa virulence in P. aeruginosa and C. albicans colonized mice. Together, our findings provide insight into how a bacterial-fungal interaction can modulate bacterial virulence in the intestine. Previously described bacterial-fungal antagonistic interactions have focused on growth inhibition or colonization inhibition/modulation, yet here we describe a novel observation of fungal-inhibition of bacterial effectors critical for virulence but not important for colonization. These findings validate the use of a mammalian model system to explore the complexities of polymicrobial, polykingdom infections in order to identify new therapeutic targets for preventing microbial disease.

  20. Characterization of a heme-regulated non-coding RNA encoded by the prrF locus of Pseudomonas aeruginosa.

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    Amanda G Oglesby-Sherrouse

    Full Text Available Pseudomonas aeruginosa, an opportunistic pathogen, requires iron for virulence and can obtain this nutrient via the acquisition of heme, an abundant source of iron in the human body. A surplus of either iron or heme can lead to oxidative stress; thus, the Fur (ferric uptake regulator protein blocks expression of genes required for iron and heme uptake in iron-replete environments. Fur also represses expression of two nearly identical genes encoding the 116- and 114-nucleotide (nt long PrrF1 and PrrF2 RNAs, respectively. While other Pseudomonads encode for the two PrrF RNAs at separate genomic loci, PrrF1 and PrrF2 are encoded in tandem in all sequenced strains of P. aeruginosa. In this report we characterize a third longer transcript encoded by the prrF locus, PrrH, which is repressed by heme as well as iron. We mapped the PrrH RNA in PA01 using 5' rapid amplification of cDNA ends (RACE and northern analysis, demonstrating the PrrH RNA is 325 nt in length. Accordingly, transcription of PrrH initiates at the 5' end of prrF1, proceeds through the prrF1 terminator and prrF1-prrF2 intergenic sequence (95 nt, and terminates at the 3' end of the prrF2 gene. We also present evidence that repression of PrrH by heme causes increased expression of previously identified PrrF-regulated genes, as well as newly identified iron- and heme-activated genes. Thus, the PrrH RNA appears to impart a novel heme regulatory mechanism to P. aeruginosa.

  1. Elucidation of the 3-O-deacylase gene, pagL, required for the removal of primary β-hydroxy fatty acid from the lipid A in the nitrogen-fixing endosymbiont Rhizobium etli CE3.

    Science.gov (United States)

    Brown, Dusty B; Muszynski, Artur; Salas, Omar; Speed, Kacie; Carlson, Russell W

    2013-04-26

    Until now, the gene responsible for the 3-O-deacylation of lipid A among nitrogen-fixing endosymbionts has not been characterized. Several Gram-negative animal pathogens such as Salmonella enterica, Pseudomonas aeruginosa, and Bordetella bronchiseptica contain an outer membrane 3-O-deacylase (PagL) that has been implicated in host immune evasion. The role of 3-O-deacylated lipid A among nitrogen-fixing endosymbionts, plant endophytes, and plant pathogens has not been studied. However, D'Haeze et al. (D'Haeze, W., Leoff, C., Freshour, G., Noel, K. D., and Carlson, R. W. (2007) J. Biol. Chem. 282, 17101-17113) reported that the lipopolysaccharide from Rhizobium etli CE3 bacteroids isolated from host bean root nodules contained exclusively tetraacylated lipid A that lacked a lipid A β-hydroxymyristyl residue, an observation that is consistent with the possibility of PagL activity being important in symbiosis. A putative pagL gene was identified in the R. etli genome sequence. With this information, we created a pagL(-) mutant strain derived from R. etli CE3. Using mass spectrometry, we demonstrated that the mutant lacks 3-O-deacylated lipid A. The parent and mutant LPS were very similar as determined by gel electrophoresis and glycosyl composition analysis using gas chromatography/mass spectrometry. However, fatty acid analysis showed that the mutant lipid A contained larger amounts of β-hydroxypentadecanoic acid than that of the parent. Furthermore, the mutant was adversely affected in establishing symbiosis with its host, Phaseolus vulgaris.

  2. Phenotypic characterization and PCR-Ribotypic profile of Pseudomonas aeruginosa isolated from cystic fibrosis patients in Iran

    Directory of Open Access Journals (Sweden)

    Hossein Fazeli

    2013-01-01

    Conclusion: The isolates of P. aeruginosa showed meaningful difference between drug resistance to antibiotics. The majority of P. aeruginosa isolated from CF patients showed pattern1 of PCR-Ribotyping.

  3. Identification of outer membrane Porin D as a vitronectin-binding factor in cystic fibrosis clinical isolates of Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Paulsson, Magnus; Singh, Birendra; Al-Jubair, Tamim;

    2015-01-01

    BACKGROUND: Pseudomonas aeruginosa is a pathogen that frequently colonizes patients with cystic fibrosis (CF) or chronic obstructive pulmonary disease (COPD). Several pathogens are known to bind vitronectin to increase their virulence. Vitronectin has been shown to enhance P. aeruginosa adhesion ...

  4. 8 CFR 216.2 - Notification requirements.

    Science.gov (United States)

    2010-01-01

    ... requirement that the alien and the petitioning spouse or alien entrepreneur must file a petition to remove the... petitioning spouse, or alien entrepreneur of the requirement to file a petition to remove conditions...

  5. Pseudomonas aeruginosa KUCd1, a possible candidate for cadmium bioremediation Pseudomonas aeruginosa KUCd1, um possível candidato para biorremediação de cádmio

    Directory of Open Access Journals (Sweden)

    Sangram Sinha

    2009-09-01

    Full Text Available A cadmium (8 mM resistant Pseudomonas aeruginosa strain KUCd1 exhibiting high Cd accumulation under in vitro aerobic condition has been reported. The isolate showed a significant ability to remove more than 75% and 89% of the soluble cadmium during the active growth phase from the growth medium and from Cd-amended industrial wastewater under growth supportive condition. Transmission electron microscopy (TEM and energy dispersive X-ray spectroscopy (EDXS suggest the presence of Cd in the cells from mid-stationary phase. The cell fractionation study revealed membrane and periplasm to be the major accumulating site in this strain. The chemical nature of the accumulated Cd was studied by X-ray powder diffraction analysis.Descreve-se a cepa Pseudomonas aeruginosa KUCd1 resistente a cádmio (8mM que apresenta elevado acumulo de cádmio em condições de aerobiose in vitro. Durante a fase ativa de multiplicação, a cepa apresentou capacidade de remover mais de 75% e 89% de cádmio solúvel do meio de cultura e da água residual industrial contendo Cd. A microscopia eletrônica de transmissão (TEM e espectroscopia de raio X energia dispersiva indicaram a presença de Cd nas células na fase estacionária intermediária. O fracionamento celular indicou serem a membrana e o periplasma os pontos de maior acúmulo. A natureza química do Cd acumulado foi avaliada através de análise de difração de Raios X.

  6. Oral biofilm models for mechanical plaque removal

    NARCIS (Netherlands)

    Verkaik, Martinus J.; Busscher, Henk J.; Rustema-Abbing, Minie; Slomp, Anje M.; Abbas, Frank; van der Mei, Henny C.

    2010-01-01

    In vitro plaque removal studies require biofilm models that resemble in vivo dental plaque. Here, we compare contact and non-contact removal of single and dual-species biofilms as well as of biofilms grown from human whole saliva in vitro using different biofilm models. Bacteria were adhered to a sa

  7. Antibiotic susceptibility patterns of Pseudomonas aeruginosa at a tertiary care hospital in Gujarat, India

    Directory of Open Access Journals (Sweden)

    Javiya Viren

    2008-01-01

    Full Text Available Objectives: The present study was undertaken to assess the antibiotic susceptibility patterns of Pseudomonas aeruginosa at a tertiary care hospital in Gujarat, India. Due to significant changes in microbial genetic ecology, as a result of indiscriminate use of anti-microbials, the spread of anti-microbial resistance is now a global problem. Materials and Methods: Out of 276 culture positive samples, 56 samples of Pseudomonas aeruginosa were examined and 10 different types of specimen were collected. Microbial sensitivity testing was done using disk diffusion test with Pseudomonas species NCTC 10662, as per CLSI guidelines. Results: The highest number of Pseudomonas infections was found in urine, followed by pus and sputum. Pseudomonas species demonstrated marked resistance against monotherapy of penicillins, cephalosporins, fluoroquinolones, tetracyclines and macrolides. Only combination drugs like Ticarcillin + Clavulanic acid, Piperacillin + Tazobactum, Cefoperazone + Sulbactum, Cefotaxime + Sulbactum, Ceftriaxome + Sulbactum and monotherapy of amikacin showed higher sensitivity to Pseudomonas infections; however, the maximum sensitivity was shown by the Carbapenems. Conclusion: From the present study, we conclude that urinary tract infection was the most common hospital acquired infection. Also, co-administration of β -lactamase inhibitors markedly expanded the anti-microbial sensitivity of semi-synthetic penicillins and cephalosporins. The aminoglycoside group of antibiotics - amikacin - demonstrated maximum sensitivity against pseudomonas species. Therefore, use of amikacin should be restricted to severe nosocomial infections, in order to avoid rapid emergence of resistant strains. Periodic susceptibility testing should be carried out over a period of two to three years, to detect the resistance trends. Also, a rational strategy on the limited and prudent use of anti-Pseudomonal agents is urgently required.

  8. FimL regulates cAMP synthesis in Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Yuki F Inclan

    Full Text Available Pseudomonas aeruginosa, a ubiquitous bacteria found in diverse ecological niches, is an important cause of acute infections in immunocompromised individuals and chronic infections in patients with Cystic Fibrosis. One signaling molecule required for the coordinate regulation of virulence factors associated with acute infections is 3', 5'-cyclic adenosine monophosphate, (cAMP, which binds to and activates a catabolite repressor homolog, Vfr. Vfr controls the transcription of many virulence factors, including those associated with Type IV pili (TFP, the Type III secretion system (T3SS, the Type II secretion system, flagellar-mediated motility, and quorum sensing systems. We previously identified FimL, a protein with histidine phosphotransfer-like domains, as a regulator of Vfr-dependent processes, including TFP-dependent motility and T3SS function. In this study, we carried out genetic and physiologic studies to further define the mechanism of action of FimL. Through a genetic screen designed to identify suppressors of FimL, we found a putative cAMP-specific phosphodiesterase (CpdA, suggesting that FimL regulates cAMP levels. Inactivation of CpdA increases cAMP levels and restores TFP-dependent motility and T3SS function to fimL mutants, consistent with in vivo phosphodiesterase activity. By constructing combinations of double and triple mutants in the two adenylate cyclase genes (cyaA and cyaB, fimL, and cpdA, we show that ΔfimL mutants resemble ΔcyaB mutants in TM defects, decreased T3SS transcription, and decreased cAMP levels. Similar to some of the virulence factors that they regulate, we demonstrate that CyaB and FimL are polarly localized. These results reveal new complexities in the regulation of diverse virulence pathways associated with acute P. aeruginosa infections.

  9. A conformational landscape for alginate secretion across the outer membrane of Pseudomonas aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Tan, Jingquan [Trinity College, Dublin (Ireland); Rouse, Sarah L. [University of Oxford, South Parks Road, Oxford (United Kingdom); Li, Dianfan; Pye, Valerie E.; Vogeley, Lutz; Brinth, Alette R.; El Arnaout, Toufic [Trinity College, Dublin (Ireland); Whitney, John C.; Howell, P. Lynne [The Hospital for Sick Children, Toronto, Ontario (Canada); University of Toronto, Toronto, Ontario (Canada); Sansom, Mark S. P. [University of Oxford, South Parks Road, Oxford (United Kingdom); Caffrey, Martin, E-mail: martin.caffrey@tcd.ie [Trinity College, Dublin (Ireland)

    2014-08-01

    Crystal structures of the β-barrel porin AlgE reveal a mechanism whereby alginate is exported from P. aeruginosa for biofilm formation. The exopolysaccharide alginate is an important component of biofilms produced by Pseudomonas aeruginosa, a major pathogen that contributes to the demise of cystic fibrosis patients. Alginate exits the cell via the outer membrane porin AlgE. X-ray structures of several AlgE crystal forms are reported here. Whilst all share a common β-barrel constitution, they differ in the degree to which loops L2 and T8 are ordered. L2 and T8 have been identified as an extracellular gate (E-gate) and a periplasmic gate (P-gate), respectively, that reside on either side of an alginate-selectivity pore located midway through AlgE. Passage of alginate across the membrane is proposed to be regulated by the sequential opening and closing of the two gates. In one crystal form, the selectivity pore contains a bound citrate. Because citrate mimics the uronate monomers of alginate, its location is taken to highlight a route through AlgE taken by alginate as it crosses the pore. Docking and molecular-dynamics simulations support and extend the proposed transport mechanism. Specifically, the P-gate and E-gate are flexible and move between open and closed states. Citrate can leave the selectivity pore bidirectionally. Alginate docks stably in a linear conformation through the open pore. To translate across the pore, a force is required that presumably is provided by the alginate-synthesis machinery. Accessing the open pore is facilitated by complex formation between AlgE and the periplasmic protein AlgK. Alginate can thread through a continuous pore in the complex, suggesting that AlgK pre-orients newly synthesized exopolysaccharide for delivery to AlgE.

  10. Eradication of Pseudomonas aeruginosa biofilms by atmospheric pressure non-thermal plasma.

    Directory of Open Access Journals (Sweden)

    Mahmoud Y Alkawareek

    Full Text Available Bacteria exist, in most environments, as complex, organised communities of sessile cells embedded within a matrix of self-produced, hydrated extracellular polymeric substances known as biofilms. Bacterial biofilms represent a ubiquitous and predominant cause of both chronic infections and infections associated with the use of indwelling medical devices such as catheters and prostheses. Such infections typically exhibit significantly enhanced tolerance to antimicrobial, biocidal and immunological challenge. This renders them difficult, sometimes impossible, to treat using conventional chemotherapeutic agents. Effective alternative approaches for prevention and eradication of biofilm associated chronic and device-associated infections are therefore urgently required. Atmospheric pressure non-thermal plasmas are gaining increasing attention as a potential approach for the eradication and control of bacterial infection and contamination. To date, however, the majority of studies have been conducted with reference to planktonic bacteria and rather less attention has been directed towards bacteria in the biofilm mode of growth. In this study, the activity of a kilohertz-driven atmospheric pressure non-thermal plasma jet, operated in a helium oxygen mixture, against Pseudomonas aeruginosa in vitro biofilms was evaluated. Pseudomonas aeruginosa biofilms exhibit marked susceptibility to exposure of the plasma jet effluent, following even relatively short (≈ 10's s exposure times. Manipulation of plasma operating conditions, for example, plasma operating frequency, had a significant effect on the bacterial inactivation rate. Survival curves exhibit a rapid decline in the number of surviving cells in the first 60 seconds followed by slower rate of cell number reduction. Excellent anti-biofilm activity of the plasma jet was also demonstrated by both confocal scanning laser microscopy and metabolism of the tetrazolium salt, XTT, a measure of bactericidal

  11. Role of ppGpp in Pseudomonas aeruginosa acute pulmonary infection and virulence regulation.

    Science.gov (United States)

    Xu, Xiaohui; Yu, Hua; Zhang, Di; Xiong, Junzhi; Qiu, Jing; Xin, Rong; He, Xiaomei; Sheng, Halei; Cai, Wenqiang; Jiang, Lu; Zhang, Kebin; Hu, Xiaomei

    2016-11-01

    During infection, bacteria might generate adaptive responses to facilitate their survival and colonization in the host environment. The alarmone guanosine 5'-triphosphate-3'-diphosphate (ppGpp), the levels of which are regulated by the RelA and SpoT enzymes, plays a critical role in mediating bacterial adaptive responses and virulence. However, the mechanism by which ppGpp regulates virulence-associated traits in Pseudomonas aeruginosa is poorly understood. To investigate the regulatory role of ppGpp, the ppGpp-deficient strain ΔRS (relA and spoT gene double mutant) and the complemented strain ΔRS(++) (complemented with relA and spoT genes) were constructed. Herein, we reported that the ΔRS strain showed decreased cytotoxicity towards A549 human alveolar adenocarcinoma cell lines and led to reduced mortality, lung edema and inflammatory cell infiltration in a mouse model of acute pneumonia compared to wild-type PAO1 and the complemented strain ΔRS(++). Subsequent analyses demonstrated that the ΔRS strain displayed reduced T3SS expression, decreased levels of elastase activity, pyocyanin, pyoverdin and alginate, and inhibited swarming and biofilm formation compared to PAO1 and the complemented strain ΔRS(++). In addition, the results demonstrate that ppGpp-mediated regulation of T3SS, virulence factor production, and swarming occurs in a quinolone quorum-sensing system-dependent manner. Taken together, these results suggest that ppGpp is required for virulence regulation in P. aeruginosa, providing new clues for the development of interference strategies against bacterial infection.

  12. Antibiofilm activities of certain biocides in Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    S Gharavi

    2009-12-01

    Full Text Available Background and objectives: Pseudomonas aeruginosa is an opportunistic pathogen that can produce biofilm. Biofilm is a complex, three dimensional structure in which microorganisms are attached to a surface and embedded in a matrix made of extracellular polymers. Due to high resistance to antimicrobial agents, biofilms create difficulties in various situations in healthcare. In this study, antibiofilm activities of some biocides in P. aeruginosa were studied."nMaterials and methods: The biofilm production ability of P. aeruginosa strain 214 (a clinical isolate was determined in the presence of six biocides including of ethylene diamine tetra acetic acid (EDTA, silver nitrate (AgNO3, bismuth ethanedithiol (BisEDT, bismuth dimercaprol (BisBAL, bismuth-2-mercaptoethanol (BisMEO and bismuth propanedithiol (BisPDT using the modified microtiter plate method. Bactericidal activity of the biocides against biofilm and planktonic cells was investigated. In this study, permeation of biocides through alginate layer was evaluated with a sandwich cup method."nResults: The results demonstrated that in the presence of bismuth thiols, biofilm production in MIC and sub MIC concentrations was considerably inhibited. Bismuththiols had lower antibiofilm bactericidal activity than EDTA and silver nitrate. One possible mechanism of biofilm resistance is exopolysaccharide production which prevents the access of antimicrobial agents to cells inside the biofilm. Bismuth thiols could not penetrate, while EDTA and silver nitrate had high penetration rate."nConclusions: Due to the frequent use of silver nitrate and EDTA in various applications, low efficacy in the inhibition of biofilm production, unstudied toxicity of BTs for humans and high efficacy in the inhibition of biofilm production, it is suggested that combinatory effect of BTs with silver nitrate or EDTA on biofilms and biofilm production be investigated.

  13. Assembly and development of the Pseudomonas aeruginosa biofilm matrix.

    Directory of Open Access Journals (Sweden)

    Luyan Ma

    2009-03-01

    Full Text Available Virtually all cells living in multicellular structures such as tissues and organs are encased in an extracellular matrix. One of the most important features of a biofilm is the extracellular polymeric substance that functions as a matrix, holding bacterial cells together. Yet very little is known about how the matrix forms or how matrix components encase bacteria during biofilm development. Pseudomonas aeruginosa forms environmentally and clinically relevant biofilms and is a paradigm organism for the study of biofilms. The extracellular polymeric substance of P. aeruginosa biofilms is an ill-defined mix of polysaccharides, nucleic acids, and proteins. Here, we directly visualize the product of the polysaccharide synthesis locus (Psl exopolysaccharide at different stages of biofilm development. During attachment, Psl is anchored on the cell surface in a helical pattern. This promotes cell-cell interactions and assembly of a matrix, which holds bacteria in the biofilm and on the surface. Chemical dissociation of Psl from the bacterial surface disrupted the Psl matrix as well as the biofilm structure. During biofilm maturation, Psl accumulates on the periphery of 3-D-structured microcolonies, resulting in a Psl matrix-free cavity in the microcolony center. At the dispersion stage, swimming cells appear in this matrix cavity. Dead cells and extracellular DNA (eDNA are also concentrated in the Psl matrix-free area. Deletion of genes that control cell death and autolysis affects the formation of the matrix cavity and microcolony dispersion. These data provide a mechanism for how P. aeruginosa builds a matrix and subsequently a cavity to free a portion of cells for seeding dispersal. Direct visualization reveals that Psl is a key scaffolding matrix component and opens up avenues for therapeutics of biofilm-related complications.

  14. Pseudomonas aeruginosa mutations in lasl and rhll quorum sensing systems result in milder chronic lung infection

    DEFF Research Database (Denmark)

    Wu, H.; Song, Z.J.; Givskov, Michael Christian;

    2001-01-01

    To understand the importance of quorum sensing in chronic Pseudomonas aeruginosa lung infection, the in vivo pathogenic effects of the wild-type P aeruginosa PAO1 and its double mutant, PAO1 lasI rhlI, in which the signal-generating parts of the quorum sensing systems are defective were compared....

  15. Network-assisted investigation of virulence and antibiotic-resistance systems in Pseudomonas aeruginosa

    Science.gov (United States)

    Hwang, Sohyun; Kim, Chan Yeong; Ji, Sun-Gou; Go, Junhyeok; Kim, Hanhae; Yang, Sunmo; Kim, Hye Jin; Cho, Ara; Yoon, Sang Sun; Lee, Insuk

    2016-05-01

    Pseudomonas aeruginosa is a Gram-negative bacterium of clinical significance. Although the genome of PAO1, a prototype strain of P. aeruginosa, has been extensively studied, approximately one-third of the functional genome remains unknown. With the emergence of antibiotic-resistant strains of P. aeruginosa, there is an urgent need to develop novel antibiotic and anti-virulence strategies, which may be facilitated by an approach that explores P. aeruginosa gene function in systems-level models. Here, we present a genome-wide functional network of P. aeruginosa genes, PseudomonasNet, which covers 98% of the coding genome, and a companion web server to generate functional hypotheses using various network-search algorithms. We demonstrate that PseudomonasNet-assisted predictions can effectively identify novel genes involved in virulence and antibiotic resistance. Moreover, an antibiotic-resistance network based on PseudomonasNet reveals that P. aeruginosa has common modular genetic organisations that confer increased or decreased resistance to diverse antibiotics, which accounts for the pervasiveness of cross-resistance across multiple drugs. The same network also suggests that P. aeruginosa has developed mechanism of trade-off in resistance across drugs by altering genetic interactions. Taken together, these results clearly demonstrate the usefulness of a genome-scale functional network to investigate pathogenic systems in P. aeruginosa.

  16. Draft Genome Sequence of Beneficial Rice Rhizosphere Isolate Pseudomonas aeruginosa PUPa3

    OpenAIRE

    Uzelac, Gordana; Bertani, Iris; Kojic, Milan; Konrad H. Paszkiewicz; Studholme, David J.; Passos da Silva, Daniel; Venturi, Vittorio

    2014-01-01

    Pseudomonas aeruginosa PUPa3 is a rhizosphere-colonizing and plant growth-promoting strain isolated from the rhizosphere of rice. This strain has, however, been shown to be pathogenic in two nonmammalian infection models. Here we report the draft genome sequence of P. aeruginosa PUPa3.

  17. Virulence Genes Profile of Multidrug Resistant Pseudomonas aeruginosa Isolated from Iranian Children with UTIs

    Directory of Open Access Journals (Sweden)

    Zohreh Heidary

    2016-04-01

    Full Text Available Virulent and resistant strains Pseudomonas aeruginosa (P. aeruginosa is one of the most important cause of UTIs in pediatrics. The present study was carried to investigate the frequency of virulence factors in the multi-drug resistant strains of P. aeruginosa isolated from pediatrics hospitalized due to the UTIs. One - hundred and forty three urine samples were collected from pediatric patients suffered from UTIs. Samples were cultured and those that were P. aeruginosa positive were analyzed for the presence of putative virulence genes. Seventy one out of 143 samples (49.65% were positive for P. aeruginosa. Monthly, sex and age-dependent prevalence were seen for P. aeruginosa. Bacterial strains had the highest levels of resistance against ampicillin (95.77%, gentamicin (92.95% and ciprofloxacin (81.69%. Of 71 P. aeruginosa isolates, 12 strains were resistant to more than 9 antibiotics (16.90%. The most commonly detected virulence factors in the cases of urethral infections were exoU and plcH while those of pyelonephritis and cystitis were were exoS and lasB. Our findings should raise awareness about antibiotic resistance in hospitalized pediatrics with UTIs in Iran. Clinicians should exercise caution in prescribing antibiotics, especially in cases of UTIs. Such information can help in identifying these virulence genes as useful diagnostic markers for clinical P. aeruginosa strains isolated from UTIs.

  18. Clinical and Morphological Studies on Spontaneous Cases of Pseudomonas aeruginosa Infections in Birds

    Directory of Open Access Journals (Sweden)

    I Dinev1, S Denev2* and G Beev2

    2013-07-01

    Full Text Available Clinical, pathoanatomical, histological, and bacteriological studies were performed on broiler chickens, growing broiler parents, and growing egg layers, in three different poultry farms, after an outbreak of Pseudomonas aeruginosa infections. The method of contamination of the birds was established. Several local and systemic clinico-morphological forms of spontaneous P. aeruginosa infections in various categories of stock birds were described: cases of P. aeruginosa infection resulting from injection of contaminated vaccines; case of P. aeruginosa infections through contaminated aerosol vaccine and cases of pododermatitis, periarthritis and arthritis in broiler chickens associated with P. aeruginosa infection. In different cases mortality range between 0.5 and 50%. The results showed that apart from embryonic mortality in hatcheries, and septicemic infections in newly hatched chickens, the pathogenicity of P. aeruginosa was associated with localized and systemic lesions in this category, as well as in young and growing birds. On one hand, these results have a theoretical significance, contributing for the confirmation and expansion of the wide array of clinico-morphological forms of P. aeruginosa infections in birds. On the other hand, the knowledge on these forms has a purely practical significance in the diagnostics of P. aeruginosa infections by poultry pathologists and veterinary practitioners.

  19. Effects of sulfate on microcystin production, photosynthesis, and oxidative stress in Microcystis aeruginosa.

    Science.gov (United States)

    Chen, Lei; Gin, Karina Y H; He, Yiliang

    2016-02-01

    Increasing sulfate in freshwater systems, caused by human activities and climate change, may have negative effects on aquatic organisms. Microcystis aeruginosa (M. aeruginosa) is both a major primary producer and a common toxic cyanobacterium, playing an important role in the aquatic environment. This study first investigated the effects of sulfate on M. aeruginosa. The experiment presented here aims at analyzing the effects of sulfate on physiological indices, molecular levels, and its influencing mechanism. The results of our experiment showed that sulfate (at 40, 80, and 300 mg L(-1)) inhibited M. aeruginosa growth, increased both intracellular and extracellular toxin contents, and enhanced the mcyD transcript level. Sulfate inhibited the photosynthesis of M. aeruginosa, based on the decrease in pigment content and the down-regulation of photosynthesis-related genes after sulfate exposure. Furthermore, sulfate decreased the maximum electron transport rate, causing the cell to accumulate surplus electrons and form reactive oxygen species (ROS). Sulfate also increased the malondialdehyde (MDA) content, which showed that sulfate damaged the cytomembrane. This damage contributed to the release of intracellular toxin to the culture medium. Although sulfate increased superoxide dismutase (SOD) activities, expression of sod, and total antioxidant capacity in M. aeruginosa, it still overwhelmed the antioxidant system since the ROS level simultaneously increased, and finally caused oxidative stress. Our results indicate that sulfate has direct effects on M. aeruginosa, inhibits photosynthesis, causes oxidative stress, increases toxin production, and affects the related genes expression in M. aeruginosa.

  20. Pseudomonas aeruginosa biofilm infections in cystic fibrosis: insights into pathogenic processes and treatment strategies

    DEFF Research Database (Denmark)

    Hassett, Daniel J; Korfhagen, Thomas R; Irvin, Randall T;

    2010-01-01

    CF airway mucus can be infected by opportunistic microorganisms, notably Pseudomonas aeruginosa. Once organisms are established as biofilms, even the most potent antibiotics have little effect on their viability, especially during late-stage chronic infections. Better understanding of the mechani...... of the mechanisms used by P. aeruginosa to circumvent host defenses and therapeutic intervention strategies is critical for advancing novel treatment strategies....

  1. Pseudomonas aeruginosa septic arthritis of knee after intra-articular ozone injection.

    Science.gov (United States)

    Seyman, Derya; Ozen, Nevgun Sepin; Inan, Dilara; Ongut, Gozde; Ogunc, Dilara

    2012-07-01

    We describe a case of septic arthritis caused by Pseudomonas aeruginosa in an immunocompetent patient following intra-articular ozone injection into the knee. To the best of our knowledge, and after considering the current literature,we believe this case is unique as no other reports of septic arthritis caused by P. aeruginosa following intra-articular ozone injection has been made.

  2. Within-host microevolution of Pseudomonas aeruginosa in Italian cystic fibrosis patients

    DEFF Research Database (Denmark)

    Marvig, Rasmus Lykke; Dolce, Daniela; Madsen Sommer, Lea Mette;

    2015-01-01

    Chronic infection with Pseudomonas aeruginosa is a major cause of morbidity and mortality in cystic fibrosis (CF) patients, and a more complete understanding of P. aeruginosa within-host genomic evolution, transmission, and population genomics may provide a basis for improving intervention...

  3. Heterogeneity of biofilms formed by nonmucoid Pseudomonas aeruginosa isolates from patients with cystic fibrosis

    DEFF Research Database (Denmark)

    Lee, Baoleri; Haagensen, Janus Anders Juul; Ciofu, O.;

    2005-01-01

    Biofilms are thought to play a key role in the occurrence of lung infections by Pseudomonas aeruginosa in patients with cystic fibrosis (CF). In this study, 20 nonmucoid P. aeruginosa isolates collected during different periods of chronic infection from eight CF patients were assessed with respect...

  4. Garlic blocks quorum sensing and promotes rapid clearing of pulmonary Pseudomonas aeruginosa infections

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Jensen, Peter Østrup; Rasmussen, Thomas B;

    2005-01-01

    The opportunistic human pathogen Pseudomonas aeruginosa is the predominant micro-organism of chronic lung infections in cystic fibrosis patients. P. aeruginosa colonizes the lungs by forming biofilm microcolonies throughout the lung. Quorum sensing (QS) renders the biofilm bacteria highly tolerant...

  5. Pseudomonas aeruginosa tolerance to tobramycin, hydrogen peroxide and polymorphonuclear leukocytes is quorum-sensing dependent

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Jensen, Peter Østrup; Burmølle, Mette;

    2005-01-01

    The opportunistic human pathogen Pseudomonas aeruginosa is the predominant micro-organism of chronic lung infections in cystic fibrosis (CF) patients. P. aeruginosa colonizes the CF lungs by forming biofilm structures in the alveoli. In the biofilm mode of growth the bacteria are highly tolerant...

  6. Multiple roles of biosurfactants in structural biofilm development by Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Pamp, Sünje Johanna; Tolker-Nielsen, Tim

    2007-01-01

    . aeruginosa rhl4 mutants were defective in migration-dependent development of mushroom-shaped multicellular structures in the later phase of biofilm formation. Experiments involving three-color-coded mixed-strain P. aeruginosa biofilms demonstrated that the wild-type and rhl4 and pil4 mutant strains formed...

  7. Evaluation of a FRET-peptide substrate to predict virulence in Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Wendy E Kaman

    Full Text Available Pseudomonas aeruginosa produces a number of proteases that are associated with virulence and disease progression. A substrate able to detect P. aeruginosa-specific proteolytic activity could help to rapidly alert clinicians to the virulence potential of individual P. aeruginosa strains. For this purpose we designed a set of P. aeruginosa-specific fluorogenic substrates, comprising fluorescence resonance energy transfer (FRET-labeled peptides, and evaluated their applicability to P. aeruginosa virulence in a range of clinical isolates. A FRET-peptide comprising three glycines (3xGly was found to be specific for the detection of P. aeruginosa proteases. Further screening of 97 P. aeruginosa clinical isolates showed a wide variation in 3xGly cleavage activity. The absence of 3xGly degradation by a lasI knock out strain indicated that 3xGly cleavage by P. aeruginosa could be quorum sensing (QS-related, a hypothesis strengthened by the observation of a strong correlation between 3xGly cleavage, LasA staphylolytic activity and pyocyanin production. Additionally, isolates able to cleave 3xGly were more susceptible to the QS inhibiting antibiotic azithromycin (AZM. In conclusion, we designed and evaluated a 3xGly substrate possibly useful as a simple tool to predict virulence and AZM susceptibility.

  8. Multiple roles of Pseudomonas aeruginosa TBCF10839 PilY1 in motility, transport and infection

    DEFF Research Database (Denmark)

    Bohn, Yu-Sing Tammy; Brandes, Gudrun; Rakhimova, Elza

    2009-01-01

    Polymorphonuclear neutrophils are the most important mammalian host defence cells against infections with Pseudomonas aeruginosa. Screening of a signature tagged mutagenesis library of the non-piliated P. aeruginosa strain TBCF10839 uncovered that transposon inactivation of its pilY1 gene rendere...

  9. Chronic Pseudomonas aeruginosa infection definition: EuroCareCF Working Group report

    DEFF Research Database (Denmark)

    Pressler, T; Bohmova, C; Conway, S;

    2011-01-01

    Chronic pulmonary infection with P. aeruginosa develops in most patients with cystic fibrosis (CF); by adulthood 80% of patients are infected and chronic P. aeruginosa infection is the primary cause of increased morbidity and mortality in CF. Chronic infection is preceded by an intermittent stage...

  10. Pseudomonas aeruginosa biofilms in the respiratory tract of cystic fibrosis patients

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Jensen, Peter Østrup; Fiandaca, Mark J;

    2009-01-01

    The present study was undertaken to investigate the appearance and location of Pseudomonas aeruginosa in the cystic fibrosis (CF) lung and in sputum. Samples include preserved tissues of CF patients who died due to chronic P. aeruginosa lung infection prior to the advent of intensive antibiotic...

  11. Epistatic Mutations And Unpredictable Phenotypes In Pseudomonas Aeruginosa

    DEFF Research Database (Denmark)

    Andresen, Eva Kammer; Abou Hachem, Maher; Jelsbak, Lars

    2015-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen, able to adapt to stressful environments such as the cystic fibrosis (CF) airways. Adaptation of P. aeruginosa to the CF environment is associated with phenotypic changes, such as switch in mucoidy, antibiotic resistance and loss of virulence fa...

  12. [Use od ozone for disinfection of ships' system of water supply contaminated by Pseudomonas aeruginosa].

    Science.gov (United States)

    Rakhmanin, Iu A; Strikalenko, T V; Mokienko, A V; Stoianova, N V; Gutsel', Iu I

    1990-11-01

    Experimental substantiation is given of the use of ozone in doses, recommended for disinfection of water and ship water supply systems infected by Pseudomonas aeruginosa. The positive effect of ozonation of water supply systems infected by Pseudomonas aeruginosa was confirmed by results of field testing on ships of the Black sea marine steam-navigation.

  13. Glutathione exhibits antibacterial activity and increases tetracycline efficacy against Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    ZHANG YaNi; DUAN KangMin

    2009-01-01

    Glutathione (GSH) plays important roles in pulmonary diseases, and inhaled GSH therapy has been used to treat cystic fibrosis (CF) patients in clinical trials. The results in this report revealed that GSH altered the sensitivity of Pseudomonas aeruginosa to different antibiotics through pathways unrelated to the oxidative stress as generally perceived. In addition, GSH and its oxidized form inhibited the growth of P. Aeruginosa.

  14. Predicting the growth situation of Pseudomonas aeruginosa on agar plates and meat stuffs using gas sensors

    Science.gov (United States)

    Gu, Xinzhe; Sun, Ye; Tu, Kang; Dong, Qingli; Pan, Leiqing

    2016-12-01

    A rapid method of predicting the growing situation of Pseudomonas aeruginosa is presented. Gas sensors were used to acquire volatile compounds generated by P. aeruginosa on agar plates and meat stuffs. Then, optimal sensors were selected to simulate P. aeruginosa growth using modified Logistic and Gompertz equations by odor changes. The results showed that the responses of S8 or S10 yielded high coefficients of determination (R2) of 0.89–0.99 and low root mean square errors (RMSE) of 0.06–0.17 for P. aeruginosa growth, fitting the models on the agar plate. The responses of S9, S4 and the first principal component of 10 sensors fit well with the growth of P. aeruginosa inoculated in meat stored at 4 °C and 20 °C, with R2 of 0.73–0.96 and RMSE of 0.25–1.38. The correlation coefficients between the fitting models, as measured by electronic nose responses, and the colony counts of P. aeruginosa were high, ranging from 0.882 to 0.996 for both plate and meat samples. Also, gas chromatography–mass spectrometry results indicated the presence of specific volatiles of P. aeruginosa on agar plates. This work demonstrated an acceptable feasibility of using gas sensors—a rapid, easy and nondestructive method for predicting P. aeruginosa growth.

  15. Pseudomonas aeruginosa and the in vitro and in vivo biofilm mode of growth

    DEFF Research Database (Denmark)

    Høiby, N; Krogh Johansen, H; Moser, C

    2001-01-01

    The biofilm mode of growth is the survival strategy of environmental bacteria like Pseudomonas aeruginosa. Such P. aeruginosa biofilms also occur in the lungs of chronically infected cystic fibrosis patients, where they protect the bacteria against antibiotics and the immune response. The lung...

  16. [Removing biofilm from a endoscopic: evaluation of disinfection methods currently used].

    Science.gov (United States)

    Balsamo, Ana Cristina; Graziano, Kazuko Uchikawa; Schneider, René Peter; Antunes Junior, Manoel; Lacerda, Rúbia Aparecida

    2012-10-01

    Laboratory experimental study that compared the effectiveness of five methods of disinfection for the removal of biofilm in gastrointestinal endoscopes. New transparent tubes of polytetrafluoroethylene (Teflon®) were used as specimens to simulate the channels of flexible endoscopes. After pre-cleaning the tubes were intentionally contaminated with Pseudomonas aeruginosa and subjected to disinfection methods. As a result, none removed 100% of these biofilms. What else physically removed biofilm was 2% glutaraldehyde in an automatic processor, probably justified by the double clean, since the equipment has this phase at the beginning of your cycle. The method less effective for removing plaque and other debris was the acidic electrolytic water. These results suggest that the cleaning is most striking in the removal of biofilms that disinfection of consecutive since glutaraldehyde disinfectant by machine is more efficient, it is a fastener organic waste.

  17. The propeptide of Pseudomonas aeruginosa elastase acts an elastase inhibitor.

    Science.gov (United States)

    Kessler, E; Safrin, M

    1994-09-09

    Elastase, an extracellular protease of Pseudomonas aeruginosa, is synthesized as a preproenzyme containing a large amino-terminal propeptide. The propeptide is cleaved within the periplasm to form a noncovalent complex with the elastase moiety. The propeptide-elastase complex was purified from the cell extract of P. aeruginosa by affinity chromatography on Gly3-D-Phe-Sepharose. The purified fraction was proteolytically inactive and contained the propeptide-elastase complex as the major protein component. Activation by limited proteolysis with trypsin was associated with the disappearance of the propeptide. To correlate individual proteins in the preparation with proteolytic activity, the purified fraction was subjected to polyacrylamide gel electrophoresis under nondenaturing conditions and subsequent incubation of the separation gel over a skim milk-agarose-indicator gel. Clearing zones due to proteolysis were produced either by mature elastase (control) or the free processed periplasmic enzyme, a low level of which was present in the purified propeptide-elastase complex preparation. No clearing was evident with the propeptide-elastase complex, indicating inhibition by the bound propeptide. Proteolytic activity of mature elastase was inhibited by various Pseudomonas cell fractions. This inhibition was abolished by antipropeptide antibodies, and, as evident from immunoblotting analysis, was consistent with propeptide presence in the effective fraction, whole cell extract, cytosol, and one of the two periplasmic fractions obtained upon conversion of P. aeruginosa cells to spheroplasts. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electro-blotting of the various cell fractions onto nitrocellulose membranes followed by incubation of the membranes with elastase and subsequent probing with antielastase antibodies revealed elastase propeptide binding. This binding of mature elastase to the propeptide was prevented by antibodies to the propeptide but not

  18. The Approach to Pseudomonas aeruginosa in Cystic Fibrosis.

    Science.gov (United States)

    Talwalkar, Jaideep S; Murray, Thomas S

    2016-03-01

    There is a high prevalence of Pseudomonas aeruginosa in patients with cystic fibrosis and clear epidemiologic links between chronic infection and morbidity and mortality exist. Prevention and early identification of infection are critical, and stand to improve with the advent of new vaccines and laboratory methods. Once the organism is identified, a variety of treatment options are available. Aggressive use of antipseudomonal antibiotics is the standard of care for acute pulmonary exacerbations in cystic fibrosis, and providers must take into account specific patient characteristics when making treatment decisions related to antibiotic selection, route and duration of administration, and site of care.

  19. [Surviving Forms in Antibiotic-Treated Pseudomonas aeruginosa].

    Science.gov (United States)

    Mulyukin, A L; Kozlova, A N; Sorokin, V V; Suzina, N E; Cherdyntseva, T A; Kotova, I B; Gaponov, A M; Tutel'yan, A V; El'-Registan, G I

    2015-01-01

    Survival of bacterial populations treated with lethal doses of antibiotics is ensured by the presence of very small numbers of persister cells. Unlike antibiotic-resistant cells, antibiotic tolerance of persisters is not inheritable and reversible. The present work provides evidence supporting the hypothesis of transformation (maturation) of persisters of an opportunistic pathogen Pseudomonas aeruginosa revealed by ciprofloxacin (CF) treatment (25-100 μg/mL) into dormant cystlike cells (CLC) and non-culturable cells (NC), as was described previously for a number. of non-spore-forming bacteria. Subpopulations of type 1 and type 2 persisters, which survived antibiotic treatment and developed into dormant forms, were heterogeneous in their capacity to form colonies or microcolonies upon germination, in resistance to heating at 70 degrees C, and in cell morphology Type 1 persisters, which were formed after 1-month incubation in the stationary-phase cultures in the medium with decreased C and N concentrations, developed in several types of surviving cells, including those similar to CLC in cell morphology. In the course of 1-month incubation of type 2 persisters, which were formed in exponentially growing cultures, other types of surviving cells developed: immature CLC and L-forms. Unlike P. aeruginosa CLC formed in the control post-stationary phase cultures without antibiotic treatment, most of 1-month persisters, especially type 2 ones, were characterized by the loss of colony-forming capacity, probably due to transition into an uncultured state with relatively high numbers of live intact cells (Live/Dead test). Another survival strategy of P. aeruginosa populations was ensured by a minor subpopulation of CF-tolerant and CF-resistant cells able to grow in the form of microcolonies or regular colonies of decreased size in the presence of the antibiotic. The described P. aeruginosa dormant forms may be responsible for persistent forms in bacteria carriers and latent

  20. The Role of Exoenzyme S in Pseudomonas aeruginosa Infections

    Science.gov (United States)

    1988-12-20

    I1T-To FrIE MNP AD______ I. 10 00 THE ROLE OF EXOENZYME S IN PSEUDOMONAS AERUGINOSA INFECTIONS Ln 0 CNFINAL REPORT I BARBARA H. IGLEWSKI DARA W...well to the enzyme neutralization titers. Second, healthy individuals or patients infected with species other than P. aeruglnosa had either no/or low...fusions, a total of 8 stable clones were isolated and recloned. All 8 monoclonal antibodies reacted with S in Western blots, and 5 neutralized S enzyme

  1. Actividad antimicrobiana del OLEOZON® sobre Staphylococcus aureus y Pseudomonas aeruginosa

    OpenAIRE

    V. Curtiellas; M Gómez; O. Ledea; Fernández, I.; Sánchez, E.

    2005-01-01

    La actividad antimicrobiana de los aceites vegetales ozonizados suele atribuirse a la acción de los compuestos peroxídicos presentes en los mismos sobre las biomoléculas más sensibles al ataque oxidante, como son los lípidos insaturados y las proteínas que presentan grupos sulfidrilos (SH). Con el objetivo de caracterizar la actividad in vitro del aceite de girasol ozonizado, OLEOZON®, se realizó un estudio empleando las cepas S. aureus ATCC 25923 y P. aeruginosa ATCC 27853. Se determinaron l...

  2. Transformasi α-Pinena dengan Bakteri Pseudomonas aeruginosa ATCC 25923

    Directory of Open Access Journals (Sweden)

    Nanik Wijayati

    2014-03-01

    Full Text Available Indonesia adalah Negara utama yang memproduksi minyak atsiri di dunia. Minyak terpentin adalah minyak atsiri yang dihasilkan dari destilasi getah pinus Pinus merkusi J ungh. Et. De. Vr. Tujuan penelitian ini adalah untuk meningkatkan nilai minyak terpentin dengan mengubah kandungan utamanya, α-pinena menjadi senyawa baru menggunakan P. Aeruginosa dalam metode mikrobiologi. Minyak terpentin diambil dari Perhutani Laboratorium Jawa Tengah, dibuat dengan seri konsentrasi 0,5%, 1%, 2%, dan 4%. Minyak terpentin diinokulasi dalam suspensi P. areuginosa selama 48 jam pada suhu kamar (25-28oC. Hasilnya diekstraksi menggunakan dietil eter. Filtrat Terpentin dianalisis menggunakan GCdan IR. Hasil analisis GC menunjukkan puncak baru di konsentrasi 0,5%, 1%, dan 2%, tetapi dalam konsentrasi 4% tidak menunjukkan puncak baru. Hasil IR menunjukkan hidroksil (OH- dan C-O alkohol. Berdasarkan penelitian ini, dapat disimpulkan bahwa minyak terpentin dapat ditransformasi untuk menjadi senyawa yang mengandung gugus-OH melalui metode mikrobiologi dengan menggunakan bakteri P. aeruginosa. Indonesia is the main producer of essential oil in the world. Turpentine oil is an essential oil which is obtained from pine resin distillation of Pinus merkusi Jungh. et. De.Vr. The aim of this experiment was to increase the value of turpentine oil by changing its main content, i.e. α-pinene, into a new compound using P. aeruginosa in microbiological method. Turpentine oil was collected from Perhutani Central Java Laboratory, and was made into 0.5%; 1%; 2%; and 4% concentrations and it was inoculated in P. areuginosa suspension for 48 hours in room temperature (25°C-280C. The result was extracted using diethylether. The filtrate of turpentine was analyzed using GC and IR. The GC analysis result showed a new peak in 0.5%; 1%; and 2% concentrations, but in the 4% concentration didn’t show a new peak. The IR result showed alcohol with hydroxyl (-OH and –C–O groups. This

  3. Cleanups In My Community (CIMC) - Removals/Responses, National Layer

    Data.gov (United States)

    U.S. Environmental Protection Agency — This data layer provides access to Removal/Response sites as part of the CIMC web service. Removals are hazardous substance releases that require immediate or...

  4. Epidemiology of Pseudomonas aeruginosa in cystic fibrosis and the possible role of contamination by dental equipment

    DEFF Research Database (Denmark)

    Jensen, E T; Giwercman, B; Ojeniyi, B;

    1997-01-01

    Cystic fibrosis (CF) patients often suffer from Pseudomonas aeruginosa lung infection yet the source of this organism is not known. In order to determine whether CF patients might be contaminated with P. aeruginosa from dental equipment, a total of 103 water samples from 25 dental sessions...... in Frederiksberg Municipal Oral Health Care Service were examined. Three samples (2.9%) were positive for P. aeruginosa. Three hundred and twenty-seven water samples from 82 dental sessions from various other Municipal Oral Health Services in Denmark, attended by CF patients, were also examined. Eighteen of 327...... samples (5.5%) from nine sessions (11%) were positive for P. aeruginosa. In one case, genotypically identical (RFLP, pulsed-field gel electrophoresis) P. aeruginosa strains were found both in water from the dental equipment and in the CF patients sputum. This indicates a small risk for acquiring P...

  5. Evaluation of Enoyl-Acyl Carrier Protein Reductase Inhibitors as Pseudomonas aeruginosa Quorum-Quenching Reagents

    Directory of Open Access Journals (Sweden)

    Søren Molin

    2010-02-01

    Full Text Available Pseudomonas aeruginosa is an opportunistic pathogen which is responsible for a wide range of infections. Production of virulence factors and biofilm formation by P. aeruginosa are partly regulated by cell-to-cell communication quorum-sensing systems. Identification of quorum-quenching reagents which block the quorum-sensing process can facilitate development of novel treatment strategies for P. aeruginosa infections. We have used molecular dynamics simulation and experimental studies to elucidate the efficiencies of two potential quorum-quenching reagents, triclosan and green tea epigallocatechin gallate (EGCG, which both function as inhibitors of the enoyl-acyl carrier protein (ACP reductase (ENR from the bacterial type II fatty acid synthesis pathway. Our studies suggest that EGCG has a higher binding affinity towards ENR of P. aeruginosa and is an efficient quorum-quenching reagent. EGCG treatment was further shown to be able to attenuate the production of virulence factors and biofilm formation of P. aeruginosa.

  6. Effects of Iron on DNA Release and Biofilm Development by Pseudomonas Aeruginosa

    DEFF Research Database (Denmark)

    Yang, Liang; Barken, Kim Bundvig; Skindersø, Mette Elena;

    2007-01-01

    Extracellular DNA is one of the major matrix components in Pseudomonas aeruginosa biofilms. It functions as an intercellular connector and plays a role in stabilization of the biofilms. Evidence that DNA release in P. aeruginosa PAO1 biofilms is controlled by the las-rhl and pqs quorum......-sensing systems has been previously presented. This paper provides evidence that DNA release in P. aeruginosa PAO1 biofilms is also under iron regulation. Experiments involving cultivation of P. aeruginosa in microtitre trays suggested that pqs expression, DNA release and biofilm formation were favoured in media...... with low iron concentrations (5 mu M FeCIA and decreased with increasing iron concentrations. Experiments involving cultivation of P. aeruginosa in a flow-chamber system suggested that a high level of iron (1100 mu M FeCl3) in the medium suppressed DNA release, structural biofilm development...

  7. Bacteriophages of Pseudomonas aeruginosa: long-term prospects for use in phage therapy.

    Science.gov (United States)

    Krylov, Victor N

    2014-01-01

    Bacteria Pseudomonas aeruginosa, being opportunistic pathogens, are the major cause of nosocomial infections and, in some cases, the primary cause of death. They are virtually untreatable with currently known antibiotics. Phage therapy is considered as one of the possible approaches to the treatment of P. aeruginosa infections. Difficulties in the implementation of phage therapy in medical practice are related, for example, to the insufficient number and diversity of virulent phages that are active against P. aeruginosa. Results of interaction of therapeutic phages with bacteria in different conditions and environments are studied insufficiently. A little is known about possible interactions of therapeutic phages with resident prophages and plasmids in clinical strains in the foci of infections. This chapter highlights the different approaches to solving these problems and possible ways to expand the diversity of therapeutic P. aeruginosa phages and organizational arrangements (as banks of phages) to ensure long-term use of phages in the treatment of P. aeruginosa infections.

  8. Insights into the respiratory tract microbiota of patients with cystic fibrosis during early Pseudomonas aeruginosa colonization

    Energy Technology Data Exchange (ETDEWEB)

    Keravec, Marlene; Mounier, Jerome; Prestat , Emmanuel; Vallet, Sophie; Jansson, Janet K.; Bergaud , Gaetaqn; Rosec, Silvain; Gourious, Stephanie; Rault, Gilles; Coton, Emmanuel; Barbier, George; Hery-Arnaud, Geneveieve

    2015-08-09

    Abstract Pseudomonas aeruginosa plays a major role in cystic fibrosis (CF) progression. Therefore, it is important to understand the initial steps of P. aeruginosa infection. The structure and dynamics of CF respiratory tract microbial communities during the early stages of P. aeruginosa colonization were characterized by pyrosequencing and cloning-sequencing. The respiratory microbiota showed high diversity, related to the young age of the CF cohort (mean age 10 years). Wide inter- and intra-individual variations were revealed. A common core microbiota of 5 phyla and 13 predominant genera was found, the majority of which were obligate anaerobes. A few genera were significantly more prevalent in patients never infected by P. aeruginosa. Persistence of an anaerobic core microbiota regardless of P. aeruginosa status suggests a major role of certain anaerobes in the pathophysiology of lung infections in CF. Some genera may be potential biomarkers of pulmonary infection state.

  9. Dynamics and spatial distribution of beta-lactamase expression in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Bagge, N.; Hentzer, Morten; Andersen, Jens Bo

    2004-01-01

    The development of resistance to beta-lactam antibiotics is a problem in the treatment of chronic Pseudomonas aeruginosa infection in the lungs of patients with cystic fibrosis. The main resistance mechanism is high-level expression of the chromosomally encoded AmpC beta-lactamase of P. aeruginosa...... cells growing in biofilms. Several genes have been shown to influence the level of ampC expression, but little is known about the regulation of ampC expression in P. aeruginosa biofilms. To study the expression of ampC in P. aeruginosa biofilms, we constructed a reporter that consisted of the fusion...... of the ampC promoter to gfp(ASV) encoding an unstable version of the green fluorescent protein. In vitro biofilms of P. aeruginosa were exposed to the beta-lactam antibiotics imipenem and ceftazidime. Sub-MICs of imipenem significantly induced the monitor system of the biofilm bacteria in the peripheries...

  10. Phenol removal pretreatment process

    Science.gov (United States)

    Hames, Bonnie R.

    2004-04-13

    A process for removing phenols from an aqueous solution is provided, which comprises the steps of contacting a mixture comprising the solution and a metal oxide, forming a phenol metal oxide complex, and removing the complex from the mixture.

  11. On-Demand Removal of Bacterial Biofilms via Shape Memory Activation.

    Science.gov (United States)

    Gu, Huan; Lee, Sang Won; Buffington, Shelby Lois; Henderson, James H; Ren, Dacheng

    2016-08-24

    Bacterial biofilms are a major cause of chronic infections and biofouling; however, effective removal of established biofilms remains challenging. Here we report a new strategy for biofilm control using biocompatible shape memory polymers with defined surface topography. These surfaces can both prevent bacterial adhesion and remove established biofilms upon rapid shape change with moderate increase of temperature, thereby offering more prolonged antifouling properties. We demonstrate that this strategy can achieve a total reduction of Pseudomonas aeruginosa biofilms by 99.9% compared to the static flat control. It was also found effective against biofilms of Staphylococcus aureus and an uropathogenic strain of Escherichia coli.

  12. Impact of alginate-producing Pseudomonas aeruginosa on alveolar macrophage apoptotic cell clearance.

    Science.gov (United States)

    McCaslin, Charles A; Petrusca, Daniela N; Poirier, Christophe; Serban, Karina A; Anderson, Gregory G; Petrache, Irina

    2015-01-01

    Pseudomonas aeruginosa infection is a hallmark of lung disease in cystic fibrosis. Acute infection with P. aeruginosa profoundly inhibits alveolar macrophage clearance of apoptotic cells (efferocytosis) via direct effect of virulence factors. During chronic infection, P. aeruginosa evades host defense by decreased virulence, which includes the production or, in the case of mucoidy, overproduction of alginate. The impact of alginate on innate immunity, in particular on macrophage clearance of apoptotic cells is not known. We hypothesized that P. aeruginosa strains that exhibit reduced virulence impair macrophage clearance of apoptotic cells and we investigated if the polysaccharide alginate produced by mucoid P. aeruginosa is sufficient to inhibit alveolar macrophage efferocytosis. Rat alveolar or human peripheral blood monocyte (THP-1)-derived macrophage cell lines were exposed in vitro to exogenous alginate or to wild type or alginate-overproducing mucoid P. aeruginosa prior to challenge with apoptotic human Jurkat T-lymphocytes. The importance of LPS contamination and that of structural integrity of alginate polymers was tested using alginate of different purities and alginate lyase, respectively. Alginate inhibited alveolar macrophage efferocytosis in a dose- and time-dependent manner. This effect was augmented but not exclusively attributed to lipopolysaccharide (LPS) present in alginates. Alginate-producing P. aeruginosa inhibited macrophage efferocytosis by more than 50%. A mannuronic-specific alginate lyase did not restore efferocytosis inhibited by exogenous guluronic-rich marine alginate, but had a marked beneficial effect on efferocytosis of alveolar macrophages exposed to mucoid P. aeruginosa. Despite decreased virulence, mucoid P. aeruginosa may contribute to chronic airway inflammation through significant inhibition of alveolar clearance of apoptotic cells and debris. The mechanism by which mucoid bacteria inhibit efferocytosis may involve alginate

  13. PROGRAMMING OFFICE REMOVALS

    CERN Multimedia

    Groupe ST-HM

    2000-01-01

    The Removals Service recommends you to plan your removals well in advance, taking into account the fact that the Transport and Handling Group’s main priority remains the dismantling of LEP and the installation of the LHC. The requests can be made by: http://st.web.cern.ch/st/hm/removal/DEMEE.HTM Thank you for your cooperation.

  14. MECANISMOS DE RESISTENCIA EN PSEUDOMONAS AERUGINOSA: ENTENDIENDO A UN PELIGROSO ENEMIGO Resistance mechanisms in Pseudomonas aeruginosa: understanding a dangerous enemy

    Directory of Open Access Journals (Sweden)

    Carlos Andrés Gómez Álvarez

    2005-01-01

    Full Text Available Pseudomonas aeruginosa es un bacilo Gram negativo no fermentador, ampliamente relacionado con la infección nosocomial. Este tipo de infecciones se presentan en pacientes severamente comprometidos, hospitalizados especialmente en unidades de cuidado intensivo, donde existe una alta presión de selección de resistencia por parte de los antibióticos. Estas infecciones nosocomiales tienen implicaciones en el pronóstico del paciente, los costos del tratamiento, la estancia hospitalaria, la morbilidad y la mortalidad. Es importante que en cada institución hospitalaria se mantenga una estrecha vigilancia de los perfiles de resistencia de esta bacteria, con el fin de reconocer sus mecanismos de resistencia, su evolución y la forma de transferencia. En este sentido, un concepto como "la lectura interpretativa del antibiograma" se impone y ayuda al clínico a inferir los posibles mecanismos de resistencia que exhibe la bacteria para de esta manera orientar el uso de la terapia antibiótica y avanzar en el gran desafío que implica enfrentar las consecuencias de la infección por P. aeruginosa.Pseudomonas aeruginosa is a Gram-negative fermentative bacilli related with nosocomial infections. This kind of infections is more frequent in critical ill patients, specially in intensive care units, where a high pressure selection is ejerxed. Nosocomial infections are associated with poor prognosis, increased treatment cost, cubed length, morbidity and mortality. Each health care institution might establish antimicrobial resistance surveillance in order to recognize antimicrobial resistance mechanisms, and transference of resistance of this pathogen. In the other hand, concepts as "interpretative reading" help the clinician to infer the possible mechanisms involved and in this way guide the antimicrobial therapy in order to boarding the challenge of this kind of infections.

  15. Identification of Chemical Compounds That Inhibit the Function of Glutamyl-tRNA Synthetase from Pseudomonas aeruginosa.

    Science.gov (United States)

    Hu, Yanmei; Guerrero, Edgar; Keniry, Megan; Manrrique, Joel; Bullard, James M

    2015-10-01

    Pseudomonas aeruginosa glutamyl-tRNA synthetase (GluRS) was overexpressed in Escherichia coli. Sequence analysis indicated that P. aeruginosa GluRS is a discriminating GluRS and, similar to other GluRS proteins, requires the presence of tRNA(Glu) to produce a glutamyl-AMP intermediate. Kinetic parameters for interaction with tRNA were determined and the k(cat) and KM were 0.8 s(-1) and 0.68 µM, respectively, resulting in a k(cat)/KM of 1.18 s(-1) µM(-1). A robust aminoacylation-based scintillation proximity assay (SPA) assay was developed and 800 natural products and 890 synthetic compounds were screened for inhibitory activity against P. aeruginosa GluRS. Fourteen compounds with inhibitory activity were identified. IC50s were in the low micromolar range. The minimum inhibitory concentration (MIC) was determined for each of the compounds against a panel of pathogenic bacteria. Two compounds, BT_03F04 and BT_04B09, inhibited GluRS with IC50s of 21.9 and 24.9 µM, respectively, and both exhibited promising MICs against Gram-positive bacteria. Time-kill studies indicated that one compound was bactericidal and one was bacteriostatic against Gram-positive bacteria. BT_03F04 was found to be noncompetitive with both ATP and glutamic acid, and BT_04B09 was competitive with glutamic acid but noncompetitive with ATP. The compounds were not observed to be toxic to mammalian cells in MTT assays.

  16. Electrocatalytic Study of Pseudomonas Aeruginosa BTE-1 Strain%绿脓杆菌Pseudomonas aeruginosa BTE-1直接电催化特征

    Institute of Scientific and Technical Information of China (English)

    周萍; 张恩仁; 周立; 刁国旺; 牛俊乐

    2012-01-01

    研究产电绿脓杆菌P.aeruginosa BTE-1的电化学催化特征.结果表明,在厌氧条件下,P.aeruginosa BTE-1菌株不能分泌可充当电子介体的绿脓菌素,但可依靠在电极表面形成生物膜而呈现直接电催化性能.P.aeruginosaBTE-1在电极表面形成生物膜与其在特定电极电位下向电极传递电子的过程直接相关,适宜的电位为0.2 V(vs.SCE),电位过高可能会损害P.aeruginosa BTE-1细胞.室温范围内升高温度可增强P.aeruginosa BTE-1生物膜的电催化活性,但过高的温度(〉60℃)会抑制生物膜电催化活性.循环伏安曲线显示,在厌氧条件下形成的P.aerugi-nosa BTE-1生物膜,具有与典型产电菌株G.sulfurreducens相近的氧化还原电位(-0.4 V~-0.2 V,vs.SCE).P.aeruginosa BTE-1生物膜可电催化酵母抽取物和葡萄糖,但不能电催化醋酸盐.%The aim of the present study is to investigate the electrocatalytic activity of electricity-producing Pseudomonas aeruginosa BTE-1 strain under anaerobic conditions. Pseudomonas aeruginosa BTE-1 was inoculated into anaerobic three-electrode electrochemical cells, and the electrocatalyfic activity was measured at poised poten- tials. HPLC and cyclic voltammetry were used to detect potential electron mediators in solutions. Experimental resuits showed that no detectable pyocyanine was excreted by P. aeruginosa BTE-1 strain in the anaerobic electro- chemcial cells, and P. aeruginosa BTE-1 exhibited direct electrocatalytic activity through the formation of biofilm on the electrode surface which was induced by the electron transfer from the cells of P. aeruginosa BTE-1 to the electrode at poised potentials. Suitable potential for biofitm formation was found to be 0.2 V ( vs. SCE), and more positive potentials would lead to a potential harm to P. aeruginosa BTE-1 ceils. At room temperature, the electrocatalytic activity of the P. aeruginosa BTE-1 biofilm could be enhanced by increasing temperature, however

  17. Extracellular DNA-induced antimicrobial peptide resistance mechanisms in Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Shawn eLewenza

    2013-02-01

    Full Text Available Extracellular DNA (eDNA is in the environment, bodily fluids, in the matrix of biofilms, and accumulates at infection sites. Extracellular DNA can function as a nutrient source, a universal biofilm matrix component and an innate immune effector in extracellular DNA traps. In biofilms, eDNA is required for attachment, aggregation and stabilization of microcolonies. We have recently shown that eDNA can sequester divalent metal cations, which has interesting implications on antibiotic resistance. Extracellular DNA binds metal cations and thus activates the Mg2+-responsive PhoPQ and PmrAB two-component systems. In Pseudomonas aeruginosa and many other Gram-negative bacteria, the PhoPQ/PmrAB systems control various genes required for virulence and resisting killing by antimicrobial peptides, including the pmr genes (PA3552-PA3559 that are responsible for the addition of aminoarabinose to lipid A. The PA4773-PA4775 genes are a second DNA-induced cluster and are required for the production of spermidine on the outer surface, which protects the outer membrane from antimicrobial peptide treatment. Both modifications mask the negative surface charges and limit membrane damage by antimicrobial peptides. DNA-enriched biofilms or planktonic cultures have increased antibiotic resistance phenotypes to antimicrobial peptides and aminoglycosides. These dual antibiotic resistance and immune evasion strategies may be expressed in DNA-rich environments and contribute to long-term survival.

  18. Boolean network model of the Pseudomonas aeruginosa quorum sensing circuits.

    Science.gov (United States)

    Dallidis, Stylianos E; Karafyllidis, Ioannis G

    2014-09-01

    To coordinate their behavior and virulence and to synchronize attacks against their hosts, bacteria communicate by continuously producing signaling molecules (called autoinducers) and continuously monitoring the concentration of these molecules. This communication is controlled by biological circuits called quorum sensing (QS) circuits. Recently QS circuits and have been recognized as an alternative target for controlling bacterial virulence and infections without the use of antibiotics. Pseudomonas aeruginosa is a Gram-negative bacterium that infects insects, plants, animals and humans and can cause acute infections. This bacterium has three interconnected QS circuits that form a very complex and versatile QS system, the operation of which is still under investigation. Here we use Boolean networks to model the complete QS system of Pseudomonas aeruginosa and we simulate and analyze its operation in both synchronous and asynchronous modes. The state space of the QS system is constructed and it turned out to be very large, hierarchical, modular and scale-free. Furthermore, we developed a simulation tool that can simulate gene knock-outs and study their effect on the regulons controlled by the three QS circuits. The model and tools we developed will give to life scientists a deeper insight to this complex QS system.

  19. PARTIAL PURIFICATION AND CHARACTERIZATION OF ALKALOPHILIC PROTEASE FROM PSEUDOMONAS AERUGINOSA

    Directory of Open Access Journals (Sweden)

    R. Satheeskumar

    2013-10-01

    Full Text Available Partial purification and characterization of alkalophilic protease production from Pseudomonas aeruginosa was isolated from the gut of marine and coastal waters shrimp Penaeus monodon. The protease production was assayed in submerged fermentation to produce maximum protease activity (423 ± 0.09 U/ml. The enzyme was precipitated with ammonium sulphate and partially purified by ion exchange chromatography through DEAE Sephadex A-50 column. In 10th fraction showed maximum protease activity (734 ± 0.18 U/ml with increase in purification fold. The molecular weight of protease from Pseudomonas aeruginosa was recorded as 60 kDa. The stability of protease was tested at various pH and temperature; it showed maximum protease activity at pH-9 and temperature 50ºC. Among the various surfactants tested for enzyme stability, maximum activity was retained in poly ethylene glycol. The compatibility of protease enzyme with various commercial detergents; the enzyme retained maximum protease activity in tide. The results are indicated that all these properties make the bacterial proteases are most suitable for wide industrial applications.

  20. Fructooligosacharides reduce Pseudomonas aeruginosa PAO1 pathogenicity through distinct mechanisms.

    Science.gov (United States)

    Ortega-González, Mercedes; Sánchez de Medina, Fermín; Molina-Santiago, Carlos; López-Posadas, Rocío; Pacheco, Daniel; Krell, Tino; Martínez-Augustin, Olga; Abdelali, Daddaoua

    2014-01-01

    Pseudomonas aeruginosa is ubiquitously present in the environment and acts as an opportunistic pathogen on humans, animals and plants. We report here the effects of the prebiotic polysaccharide inulin and its hydrolysed form FOS on this bacterium. FOS was found to inhibit bacterial growth of strain PAO1, while inulin did not affect growth rate or yield in a significant manner. Inulin stimulated biofilm formation, whereas a dramatic reduction of the biofilm formation was observed in the presence of FOS. Similar opposing effects were observed for bacterial motility, where FOS inhibited the swarming and twitching behaviour whereas inulin caused its stimulation. In co-cultures with eukaryotic cells (macrophages) FOS and, to a lesser extent, inulin reduced the secretion of the inflammatory cytokines IL-6, IL-10 and TNF-α. Western blot experiments indicated that the effects mediated by FOS in macrophages are associated with a decreased activation of the NF-κB pathway. Since FOS and inulin stimulate pathway activation in the absence of bacteria, the FOS mediated effect is likely to be of indirect nature, such as via a reduction of bacterial virulence. Further, this modulatory effect is observed also with the highly virulent ptxS mutated strain. Co-culture experiments of P. aeruginosa with IEC18 eukaryotic cells showed that FOS reduces the concentration of the major virulence factor, exotoxin A, suggesting that this is a possible mechanism for the reduction of pathogenicity. The potential of these compounds as components of antibacterial and anti-inflammatory cocktails is discussed.

  1. Bacteriophages for the treatment of Pseudomonas aeruginosa infections.

    Science.gov (United States)

    Harper, D R; Enright, M C

    2011-07-01

    Bacteriophages were first identified in 1915 and were used as antimicrobial agents from 1919 onwards. Despite apparent successes and widespread application, early users did not understand the nature of these agents and their efficacy remained controversial. As a result, they were replaced in the west by chemical antibiotics once these became available. However, bacteriophages remained a common therapeutic approach in parts of Eastern Europe where they are still in use. Increasing levels of antibiotic-resistant bacterial infections are now driving demand for novel therapeutic approaches. In cases where antibiotic options are limited or nonexistent, the pressure for new agents is greatest. One of the most prominent areas of concern is multidrug-resistant Gram-negative bacteria. Pseudomonas aeruginosa is a prominent member of this class and is the cause of damaging infections that can be resistant to successful treatment with conventional antibiotics. At the same time, it exhibits a number of properties that make it a suitable target for bacteriophage-based approaches, including growth in biofilms that can hydrolyse following phage infection. Pseudomonas aeruginosa provides a striking example of an infection where clinical need and the availability of a practical therapy coincide.

  2. [Pseudomonas aeruginosa bacteriaemia: new clinical and therapeutic aspects ].

    Science.gov (United States)

    Janbon, F; Despaux, E; Lepeu, G; Jonquet, O; Santoni, A; Balmayer, B; Bertrand, A

    1982-06-01

    Fifty one cases of Pseudomonas aeruginosa bacteriaemia observed during the last 12 years are reported. Thirty five patients were over fifty years old; 92 p. cent were admitted for several days and about 50 p. cent were in post-operative period. A previous antibiotherapy and an impaired status are promotive factors. The respiratory or peritoneal origins are the most frequent. All patients were feverish; 24 have had an infectious shock which was inaugural in 12 cases. Seven pneumonitis, 3 endocarditis, one pericarditis and 2 osteitis were observed. An ecthyma gangrenosum was noted in three patients. Mortality was 70 p. cent. Comparison between recovered and died patients improved bad prognosis of old age, post operative period, neoplasic, previous organica weakness and pulmonary or peritoneal origins. Used alone, colimycin has seemed to be more effective than aminosid antibiotics; but their association with betalactamins was better. An in vitro study of the susceptibility of 100 Pseudomonas aeruginosa strains has proved the interest of piperacillin and cefsulodin; azlocillin, cefoperazone and ceftriaxone are just less effective.

  3. Identification of Pseudomonas aeruginosa genes associated with antibiotic susceptibility

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Pseudomonas aeruginosa causes acute and chronic infections in humans and these infections are difficult to treat due to the bacteria’s high-level of intrinsic and acquired resistance to antibiotics. To address this problem, it is crucial to investigate the molecular mechanisms of antibiotic resistance in this organism. In this study, a P. aeruginosa transposon insertion library of 17000 clones was constructed and screened for altered susceptibility to seven antibiotics. Colonies grown on agar plates con- taining antibiotics at minimum inhibitory concentrations (MICs) and those unable to grow at ? MIC were collected. The transposon-disrupted genes in 43 confirmed mutants that showed at least a three-fold increase or a two-fold decrease in suscep- tibility to at least one antibiotic were determined by semi-random PCR and subsequent sequencing analysis. In addition to nine genes known to be associated with antibiotic resistance, including mexI, mexB and mexR, 24 new antibiotic resis- tance-associated genes were identified, including a fimbrial biogenesis gene pilY1 whose disruption resulted in a 128-fold in- crease in the MIC of carbenicillin. Twelve of the 43 genes identified were of unknown function. These genes could serve as targets to control or reverse antibiotic resistance in this important human pathogen.

  4. [Allelopathy effects of ferulic acid and coumarin on Microcystis aeruginosa].

    Science.gov (United States)

    Guo, Ya-Li; Fu, Hai-Yan; Huang, Guo-He; Gao, Pan-Feng; Chai, Tian; Yan, Bin; Liao, Huan

    2013-04-01

    The inhibitory effects and allelopathy mechanism of ferulic acid and coumarin on Microcystis aeruginosa were investigated by measuring the D680 value, the content of chlorophyll-a, the electrical conductivity (EC) and superoxide anion radical O*- value. Ferulic acid and coumarin had allelopathic effects on the growth of M. aeruginosa and promoted the physiological metabolism at low concentrations while inhibited the metabolism at high concentrations. Obvious inhibitory effects were observed when the concentration of ferulic acid or coumarin was over 100 mg x L(-1). The average inhibitory rates reached 80.3% and 58.0% after six days when the concentration of ferulic acid or coumarin was 200 mg x L(-1). The content of chlorophyll-a was decreased while the EC value and O2*- concentration were promoted by higher concentrations of ferulic acid or coumarin, suggesting that the growth of algae was inhibited probably by the damage of cell membrane, increase in the content of O2*- and decrease in the content of chlorophyll-a. In addition, seed germination test elucidated that Ferulic acid was safer than Coumarin.

  5. Effect of methylglyoxal on multidrug-resistant Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Katsuhiko eHayashi

    2014-04-01

    Full Text Available Honey has a complex chemistry, and its broad-spectrum antimicrobial activity varies with floral source, climate, and harvesting conditions. Methylglyoxal was identified as the dominant antibacterial component of manuka honey. Although it has been known that methylglyoxal has antibacterial activity against gram-positive bacteria, including methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus, there is not much information describing its activity against gram-negative bacteria. In this study, we report the effect of methylglyoxal against multidrug-resistant Pseudomonas aeruginosa (MDRP using 53 clinically isolated strains. We also assessed the effect of deleting the five multidrug efflux systems in P. aeruginosa, as well as the efflux systems in Escherichia coli and Salmonella enterica serovar Typhimurium, on MICs of methylglyoxal. Our results indicate that methylglyoxal inhibits the growth of MDRP at concentrations of 128–512 µg/ml (1.7–7.1 mM and is not recognized by drug efflux systems.

  6. Cyanide production by Pseudomonas fluorescens and Pseudomonas aeruginosa.

    Science.gov (United States)

    Askeland, R A; Morrison, S M

    1983-06-01

    Of 200 water isolates screened, five strains of Pseudomonas fluorescens and one strain of Pseudomonas aeruginosa were cyanogenic. Maximum cyanogenesis by two strains of P. fluorescens in a defined growth medium occurred at 25 to 30 degrees C over a pH range of 6.6 to 8.9. Cyanide production per cell was optimum at 300 mM phosphate. A linear relationship was observed between cyanogenesis and the log of iron concentration over a range of 3 to 300 microM. The maximum rate of cyanide production occurred during the transition from exponential to stationary growth phase. Radioactive tracer experiments with [1-14C]glycine and [2-14C]glycine demonstrated that the cyanide carbon originates from the number 2 carbon of glycine for both P. fluorescens and P. aeruginosa. Cyanide production was not observed in raw industrial wastewater or in sterile wastewater inoculated with pure cultures of cyanogenic Pseudomonas strains. Cyanide was produced when wastewater was amended by the addition of components of the defined growth medium.

  7. Inactivation of Pseudomonas aeruginosa biofilm by dense phase carbon dioxide.

    Science.gov (United States)

    Mun, Sungmin; Jeong, Jin-Seong; Kim, Jaeeun; Lee, Youn-Woo; Yoon, Jeyong

    2009-01-01

    Dense phase carbon dioxide (DPCD) is one of the most promising techniques available to control microorganisms as a non-thermal disinfection method. However, no study on the efficiency of biofilm disinfection using DPCD has been reported. The efficiency of DPCD in inactivating Pseudomonas aeruginosa biofilm, which is known to have high antimicrobial resistance, was thus investigated. P. aeruginosa biofilm, which was not immersed in water but was completely wet, was found to be more effectively inactivated by DPCD treatment, achieving a 6-log reduction within 7 min. The inactivation efficiency increased modestly with increasing pressure and temperature. This study also reports that the water-unimmersed condition is one of the most important operating parameters in achieving efficient biofilm control by DPCD treatment. In addition, observations by confocal laser scanning microscopy revealed that DPCD treatment not only inactivated biofilm cells on the glass coupons but also caused detachment of the biofilm following weakening of its structure as a result of the DPCD treatment; this is an added benefit of DPCD treatment.

  8. Hot Spot Removal System: System description

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-09-01

    Hazardous wastes contaminated with radionuclides, chemicals, and explosives exist across the Department of Energy complex and need to be remediated due to environmental concerns. Currently, an opportunity is being developed to dramatically reduce remediation costs and to assist in the acceleration of schedules associated with these wastes by deploying a Hot Spot Removal System. Removing the hot spot from the waste site will remove risk driver(s) and enable another, more cost effective process/option/remedial alternative (i.e., capping) to be applied to the remainder of the site. The Hot Spot Removal System consists of a suite of technologies that will be utilized to locate and remove source terms. Components of the system can also be used in a variety of other cleanup activities. This Hot Spot Removal System Description document presents technologies that were considered for possible inclusion in the Hot Spot Removal System, technologies made available to the Hot Spot Removal System, industrial interest in the Hot Spot Removal System`s subsystems, the schedule required for the Hot Spot Removal System, the evaluation of the relevant technologies, and the recommendations for equipment and technologies as stated in the Plan section.

  9. A comparative study of different strategies for removal of endotoxins from bacteriophage preparations.

    Science.gov (United States)

    Van Belleghem, Jonas D; Merabishvili, Maya; Vergauwen, Bjorn; Lavigne, Rob; Vaneechoutte, Mario

    2017-01-01

    Bacterial endotoxins have high immunogenicity. Phage biology studies as well as therapeutic phage applications necessitate highly purified phage particles. In this study, we compared combinations of seven different endotoxin removal strategies and validated their endotoxin removal efficacy for five different phages (i.e. four Pseudomonas aeruginosa phages and one Staphylococcus aureus phage). These purification strategies included Endotrap HD column purification and/or CsCl density centrifugation in combination with Endotrap purification, followed by organic solvent (1-octanol), detergent (Triton X-100), enzymatic inactivation of the endotoxin using alkaline phosphatase and CIM monolytic anion exchange chromatography. We show that CsCl density purification of the P. aeruginosa phages, at an initial concentration of 10(12)-10(13)pfu/ml, led to the strongest reduction of endotoxins, with an endotoxin removal efficacy of up to 99%, whereas additional purification methods did not result in a complete removal of endotoxins from the phage preparations and only yielded an additional endotoxin removal efficacy of 23 to 99%, sometimes accompanied with strong losses in phage titer.

  10. Nosocomial outbreak of a non-cefepime-susceptible ceftazidime-susceptible Pseudomonas aeruginosa strain overexpressing MexXY-OprM and producing an integron-borne PSE-1 betta-lactamase.

    Science.gov (United States)

    Peña, C; Suarez, C; Tubau, F; Juan, C; Moya, B; Dominguez, M A; Oliver, A; Pujol, M; Ariza, J

    2009-08-01

    Cefepime (FEP) and ceftazidime (CAZ) are broad-spectrum cephalosporins that display similar MICs for wild-type Pseudomonas aeruginosa strains. Recently, P. aeruginosa isolates showing a discordance in susceptibility to CAZ and FEP have been noted at the Hospital de Bellvitge in Barcelona, Spain, and a clustering was suspected. During the study period (March to December 2007), 51 patients, particularly those in an intensive care units (ICUs) (n = 29 [57%]), infected or colonized with at least one P. aeruginosa non-FEP-susceptible and CAZ-susceptible (Fep(ns) Caz(s)) phenotype strain were detected. Twenty-three (45%) patients were infected, and the respiratory tract was the most frequent site of infection. Changes in the consumption of antimicrobials in the ICUs were observed over time: a progressive reduction in the levels of consumption of carbapenems (247 defined daily doses [DDD]/1,000 patient days to 66 DDD/1,000 patient days; P = 0.008), after restriction of its use in 2006, and an expected increase in the rate of piperacillin-tazobactam use (42 DDD/1,000 patient days in 2004 to 200 DDD/1,000 patient days in 2007; P < 0.001). Throughout the whole study period, only a single clone of a P. aeruginosa Fep(ns) Caz(s) phenotype strain was identified by pulsed-field gel electrophoresis analysis to be associated with the hyperexpression of MexXY-OprM and the production of an integron-borne PSE-1 ss-lactamase. In conclusion, we identified an epidemic P. aeruginosa clone of an Fep(ns) Caz(s) phenotype strain involving 51 patients, in particular, ICU patients. The combination of the overexpression of an efflux pump and PSE-1 ss-lactamase production is associated with the multidrug-resistant phenotype. The dominant use of a single class of antibiotics could have provided the selective pressure required for the emergence and spread of this P. aeruginosa strain.

  11. Facile biofunctionalization of silver nanoparticles for enhanced antibacterial properties, endotoxin removal, and biofilm control.

    Science.gov (United States)

    Lambadi, Paramesh Ramulu; Sharma, Tarun Kumar; Kumar, Piyush; Vasnani, Priyanka; Thalluri, Sitaramanjaneya Mouli; Bisht, Neha; Pathania, Ranjana; Navani, Naveen Kumar

    2015-01-01

    Infectious diseases cause a huge burden on healthcare systems worldwide. Pathogenic bacteria establish infection by developing antibiotic resistance and modulating the host's immune system, whereas opportunistic pathogens like Pseudomonas aeruginosa adapt to adverse conditions owing to their ability to form biofilms. In the present study, silver nanoparticles were biofunctionalized with polymyxin B, an antibacterial peptide using a facile method. The biofunctionalized nanoparticles (polymyxin B-capped silver nanoparticles, PBSNPs) were assessed for antibacterial activity against multiple drug-resistant clinical strain Vibrio fluvialis and nosocomial pathogen P. aeruginosa. The results of antibacterial assay revealed that PBSNPs had an approximately 3-fold higher effect than the citrate-capped nanoparticles (CSNPs). Morphological damage to the cell membrane was followed by scanning electron microscopy, testifying PBSNPs to be more potent in controlling the bacterial growth as compared with CSNPs. The bactericidal effect of PBSNPs was further confirmed by Live/Dead staining assays. Apart from the antibacterial activity, the biofunctionalized nanoparticles were found to resist biofilm formation. Electroplating of PBSNPs onto stainless steel surgical blades retained the antibacterial activity against P. aeruginosa. Further, the affinity of polymyxin for endotoxin was exploited for its removal using PBSNPs. It was found that the prepared nanoparticles removed 97% of the endotoxin from the solution. Such multifarious uses of metal nanoparticles are an attractive means of enhancing the potency of antimicrobial agents to control infections.

  12. SENSITIVITY TO ANTIBIOTICS, ANTISEPTICAL NOSOCOMIAL PSEUDOMONAS AERUGINOSA, ISOLATED IN UROLOGICAL PATIENTS

    Directory of Open Access Journals (Sweden)

    Rymsha E.V.

    2015-05-01

    Full Text Available Introduction. Given the active introduction into clinical practice of new groups of antibiotics and antiseptics, the problem of treatment of purulent-inflammatory complications after prostatectomy and today is relevant. Of particular concern belated cases of diagnosis and treatment of postoperative complications in urological practice patients receiving antibiotic therapy The use of traditional antibiotics is not prevents the development of infection, because the problem of resistance of microorganisms to antibiotics and antiseptics remains relevant. The solution to the problem of development of infectious complications and prevent the formation of resistant clinical strains largely depends on the isolated pathogen, susceptibility to antimicrobial agents based on its bioavailability , ability to spread and penetrate into cells and tissues, selection of dose, interval, and route of administration to maintain minimum bactericidal concentration Material and methods. The study involved 145 patients who were treated in the urology Department of the Vinnytsia regional clinical hospital named of M. I. Pirogov. Patients underwent the surgical treatment of benign hypertrophic prostate. Material for bacteriological studies of purulent-inflammatory diseases were urine, pieces of the prostate, remote operationally, urinary catheters, through which conducted irrigation of the bladder. Specimen collection, transportation was carried out in accordance with modern requirements. Identification was done by morphological, cultural and biochemical properties. The definition of antibiotic resistance were performed according to "guidelines for the definition of sensitivity of microorganisms to antibiotics by the method of diffusion in agar using discs" (No. 2675-83, Kiev, 2007 12 .]. Evaluation of the results of determining the sensitivity of microorganisms to antibiotics was carried out on the basis of the determination of the zone of growth (mm of the studied

  13. Cystic Fibrosis Transmembrane Conductance Regulator is an Epithelial Cell Receptor for Clearance of Pseudomonas aeruginosa from the Lung

    Science.gov (United States)

    Pier, Gerald B.; Grout, Martha; Zaidi, Tanweer S.

    1997-10-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride ion channel, but its relationship to the primary clinical manifestation of CF, chronic Pseudomonas aeruginosa pulmonary infection, is unclear. We report that CFTR is a cellular receptor for binding, endocytosing, and clearing P. aeruginosa from the normal lung. Murine cells expressing recombinant human wild-type CFTR ingested 30-100 times as many P. aeruginosa as cells lacking CFTR or expressing mutant Δ F508 CFTR protein. Purified CFTR inhibited ingestion of P. aeruginosa by human airway epithelial cells. The first extracellular domain of CFTR specifically bound to P. aeruginosa and a synthetic peptide of this region inhibited P. aeruginosa internalization in vivo, leading to increased bacterial lung burdens. CFTR clears P. aeruginosa from the lung, indicating a direct connection between mutations in CFTR and the clinical consequences of CF.

  14. Annona glabra Flavonoids Act As Antimicrobials by Binding to Pseudomonas aeruginosa Cell Walls

    Science.gov (United States)

    Galvão, Stanley de S. L.; Monteiro, Andrea de S.; Siqueira, Ezequias P.; Bomfim, Maria Rosa Q.; Dias-Souza, Marcus Vinícius; Ferreira, Gabriella F.; Denadai, Angelo Márcio L.; Santos, Áquila R. C.; Lúcia dos Santos, Vera; de Souza-Fagundes, Elaine M.; Fernandes, Elizabeth S.; Monteiro-Neto, Valério

    2016-01-01

    Pseudomonas aeruginosa is an important pathogen in opportunistic infections in humans. The increased incidence of antimicrobial-resistant P. aeruginosa isolates has highlighted the need for novel and more potent therapies against this microorganism. Annona glabra is known for presenting different compounds with diverse biological activities, such as anti-tumor and immunomodulatory activities. Although other species of the family display antimicrobial actions, this has not yet been reported for A. glabra. Here, we investigated the antimicrobial activity of the ethyl acetate fraction (EAF) obtained from the leaf hydroalcoholic extract of A. glabra. EAF was bactericidal against different strains of P. aeruginosa. EAF also presented with a time- and concentration-dependent effect on P. aeruginosa viability. Testing of different EAF sub-fractions showed that the sub-fraction 32-33 (SF32-33) was the most effective against P. aeruginosa. Analysis of the chemical constituents of SF32-33 demonstrated a high content of flavonoids. Incubation of this active sub-fraction with P. aeruginosa ATCC 27983 triggered an endothermic reaction, which was accompanied by an increased electric charge, suggesting a high binding of SF32-33 compounds to bacterial cell walls. Collectively, our results suggest that A. glabra-derived compounds, especially flavonoids, may be useful for treating infections caused by P. aeruginosa. PMID:28066374

  15. RNAi screen reveals an Abl kinase-dependent host cell pathway involved in Pseudomonas aeruginosa internalization.

    Directory of Open Access Journals (Sweden)

    Julia F Pielage

    2008-03-01

    Full Text Available Internalization of the pathogenic bacterium Pseudomonas aeruginosa by non-phagocytic cells is promoted by rearrangements of the actin cytoskeleton, but the host pathways usurped by this bacterium are not clearly understood. We used RNAi-mediated gene inactivation of approximately 80 genes known to regulate the actin cytoskeleton in Drosophila S2 cells to identify host molecules essential for entry of P. aeruginosa. This work revealed Abl tyrosine kinase, the adaptor protein Crk, the small GTPases Rac1 and Cdc42, and p21-activated kinase as components of a host signaling pathway that leads to internalization of P. aeruginosa. Using a variety of complementary approaches, we validated the role of this pathway in mammalian cells. Remarkably, ExoS and ExoT, type III secreted toxins of P. aeruginosa, target this pathway by interfering with GTPase function and, in the case of ExoT, by abrogating P. aeruginosa-induced Abl-dependent Crk phosphorylation. Altogether, this work reveals that P. aeruginosa utilizes the Abl pathway for entering host cells and reveals unexpected complexity by which the P. aeruginosa type III secretion system modulates this internalization pathway. Our results furthermore demonstrate the applicability of using RNAi screens to identify host signaling cascades usurped by microbial pathogens that may be potential targets for novel therapies directed against treatment of antibiotic-resistant infections.

  16. Distribution and Inhibition of Liposomes on Staphylococcus aureus and Pseudomonas aeruginosa Biofilm.

    Directory of Open Access Journals (Sweden)

    Dong Dong

    Full Text Available Staphylococcus aureus and Pseudomonas aeruginosa are major pathogens in chronic rhinosinusitis (CRS and their biofilms have been associated with poorer postsurgical outcomes. This study investigated the distribution and anti-biofilm effect of cationic (+ and anionic (- phospholipid liposomes with different sizes (unilamellar and multilamellar vesicle, ULV and MLV respectively on S. aureus and P. aeruginosa biofilms.Specific biofilm models for S. aureus ATCC 25923 and P. aeruginosa ATCC 15692 were established. Liposomal distribution was determined by observing SYTO9 stained biofilm exposed to DiI labeled liposomes using confocal scanning laser microscopy, followed by quantitative image analysis. The anti-biofilm efficacy study was carried out by using the alamarBlue assay to test the relative viability of biofilm treated with various liposomes for 24 hours and five minutes.The smaller ULVs penetrated better than larger MLVs in both S. aureus and P. aeruginosa biofilm. Except that +ULV and -ULV displayed similar distribution in S. aureus biofilm, the cationic liposomes adhered better than their anionic counterparts. Biofilm growth was inhibited at 24-hour and five-minute exposure time, although the decrease of viability for P. aeruginosa biofilm after liposomal treatment did not reach statistical significance.The distribution and anti-biofilm effects of cationic and anionic liposomes of different sizes differed in S. aureus and P. aeruginosa biofilms. Reducing the liposome size and formulating liposomes as positively charged enhanced the penetration and inhibition of S. aureus and P. aeruginosa biofilms.

  17. Behaviors of Microcystis aeruginosa cells during floc storage in drinking water treatment process

    Science.gov (United States)

    Xu, Hangzhou; Pei, Haiyan; Xiao, Hongdi; Jin, Yan; Li, Xiuqing; Hu, Wenrong; Ma, Chunxia; Sun, Jiongming; Li, Hongmin

    2016-01-01

    This is the first study to systematically investigate the different behaviors of Microcystis aeruginosa in the sludges formed by AlCl3, FeCl3, and polymeric aluminium ferric chloride (PAFC) coagulants during storage. Results show that the viability of Microcystis aeruginosa in PAFC sludge was stronger than that of cells in either AlCl3 or FeCl3 sludge after the same storage time, while the cells’ viability in the latter two systems stayed at almost the same level. In AlCl3 and FeCl3 sludges high concentrations of Al and Fe were toxic to Microcystis aeruginosa, whereas in PAFC sludge low levels of Al showed little toxic effect on Microcystis aeruginosa growth and moderate amounts of Fe were beneficial to growth. The lysis of Microcystis aeruginosa in AlCl3 sludge was more serious than that in PAFC sludge, for the same storage time. Although the cell viability in FeCl3 sludge was low (similar to AlCl3 sludge), the Microcystis aeruginosa cells remained basically intact after 10 d storage (similar to PAFC sludge). The maintenance of cellular integrity in FeCl3 sludge might be due to the large floc size and high density, which had a protective effect for Microcystis aeruginosa. PMID:27713525

  18. CHANGES IN THE MORPHOLOGY AND POLYSACCHARIDE CONTENT OF MICROCYSTIS AERUGINOSA (CYANOBACTERIA) DURING FLAGELLATE GRAZING(1).

    Science.gov (United States)

    Yang, Zhou; Kong, Fanxiang; Shi, Xiaoli; Zhang, Min; Xing, Peng; Cao, Huansheng

    2008-06-01

    To investigate the changes in the morphology and polysaccharide content of Microcystis aeruginosa (Kütz.) Kütz. during flagellate grazing, cultures of M. aeruginosa were exposed to grazing Ochromonas sp. for a period of 9 d under controlled laboratory conditions. M. aeruginosa responded actively to flagellate grazing and formed colonies, most of which were made up of several or dozens of cells, suggesting that flagellate grazing may be one of the biotic factors responsible for colony formation in M. aeruginosa. When colonies were formed, the cell surface ultrastructure changed, and the polysaccharide layer on the surface of the cell wall became thicker. This change indicated that synthesis and secretion of extracellular polysaccharide (EPS) of M. aeruginosa cells increased under flagellate grazing pressure. The contents of soluble extracellular polysaccharide (sEPS), bound extracellular polysaccharide (bEPS), and total polysaccharide (TPS) in colonial cells of M. aeruginosa increased significantly compared with those in single cells. This finding suggested that the increased amount of EPS on the cell surface may play a role in keeping M. aeruginosa cells together to form colonies.

  19. BIIL 284 reduces neutrophils numbers but increases P. aeruginosa bacteraemia and inflammation in mouse lungs

    Science.gov (United States)

    Döring, Gerd; Bragonzi, Alessandra; Paroni, Moira; Aktürk, Firdevs-Fatma; Cigana, Cristina; Schmidt, Annika; Gilpin, Deirdre; Heyder, Susanne; Born, Torsten; Smaczny, Christina; Kohlhäufl, Martin; Wagner, Thomas O. F.; Loebinger, Michael R.; Bilton, Diana; Tunney, Michael M.; Elborn, J. Stuart; Pier, Gerald B.; Konstan, Michael W.; Ulrich, Martina

    2014-01-01

    Background A clinical study to investigate the leukotriene B4 (LTB4)-receptor antagonist BIIL 284 in cystic fibrosis (CF) patients was prematurely terminated due to a significantly increased risk of adverse pulmonary events. We aimed to establish the effect of BIIL284 in models of Pseudomonas aeruginosa lung infection, thereby contributing to a better understanding of what could have led to adverse pulmonary events in CF patients. Methods P. aeruginosa DNA in the blood of CF patients during and after acute pulmonary exacerbations and in stable patients with non-CF bronchiectasis (NCFB) and healthy individuals was assessed by PCR. The effect of BIIL 284 treatment was tested in an agar beads murine model of Pseudomonas aeruginosa lung infection. Bacterial count and inflammation were evaluated in lung and other organs. Result Most CF patients (98%) and all patients with NCFB and healthy individuals had negative P. aeruginosa DNA in their blood. Similarly, the P. aeruginosa-infected mice showed bacterial counts in the lung but not blood or spleen. BIIL 284 treatment decreased pulmonary neutrophils and increased P. aeruginosa numbers in mouse lungs leading to significantly higher bacteremia rates and lung inflammation compared to placebo treated animals. Conclusions Decreased airway neutrophils induced lung proliferation and severe bacteraemia in a murine model of P. aeruginosa lung infection. These data suggest that caution should be taken when administering anti-inflammatory compounds to patients with bacterial infections. PMID:24183915

  20. Pseudomonas aeruginosa adapts its iron uptake strategies in function of the type of infections

    Directory of Open Access Journals (Sweden)

    Pierre eCornelis

    2013-11-01

    Full Text Available Pseudomonas aeruginosa is a Gram-negative -Proteobacterium which is known for its capacity to colonize various niches, including some invertebrate and vertebrate hosts, making it one of the most frequent bacteria causing opportunistic infections. P. aeruginosa is able to cause acute as well as chronic infections and it uses different colonization and virulence factors to do so. Infections range from septicemia, urinary infections, burn wound colonization, and chronic colonization of the lungs of cystic fibrosis patients. Like the vast majority of organisms, P. aeruginosa needs iron to sustain growth. P. aeruginosa utilizes different strategies to take up iron, depending on the type of infection it causes. Two siderophores are produced by this bacterium, pyoverdine and pyochelin, characterized by high and low affinities for iron respectively. P. aeruginosa is also able to utilize different siderophores from other microorganisms (siderophore piracy. It can also take up heme from hemoproteins via two different systems. Under microaerobic or anaerobic conditions, P. aeruginosa is also able to take up ferrous iron via its Feo system using redox-cycling phenazines. Depending on the type of infection, P. aeruginosa can therefore adapt by switching from one iron uptake system to another as we will describe in this short review.

  1. Solar disinfection of Pseudomonas aeruginosa in harvested rainwater: a step towards potability of rainwater.

    Directory of Open Access Journals (Sweden)

    Muhammad T Amin

    Full Text Available Efficiency of solar based disinfection of Pseudomonas aeruginosa (P. aeruginosa in rooftop harvested rainwater was evaluated aiming the potability of rainwater. The rainwater samples were exposed to direct sunlight for about 8-9 hours and the effects of water temperature (°C, sunlight irradiance (W/m2, different rear surfaces of polyethylene terephthalate bottles, variable microbial concentrations, pH and turbidity were observed on P. aeruginosa inactivation at different weathers. In simple solar disinfection (SODIS, the complete inactivation of P. aeruginosa was obtained only under sunny weather conditions (>50°C and >700 W/m2 with absorptive rear surface. Solar collector disinfection (SOCODIS system, used to improve the efficiency of simple SODIS under mild and weak weather, completely inactivated the P. aeruginosa by enhancing the disinfection efficiency of about 20% only at mild weather. Both SODIS and SOCODIS systems, however, were found inefficient at weak weather. Different initial concentrations of P. aeruginosa and/or Escherichia coli had little effects on the disinfection efficiency except for the SODIS with highest initial concentrations. The inactivation of P. aeruginosa increased by about 10-15% by lowering the initial pH values from 10 to 3. A high initial turbidity, adjusted by adding kaolin, adversely affected the efficiency of both systems and a decrease, about 15-25%; in inactivation of P. aeruginosa was observed. The kinetics of this study was investigated by Geeraerd Model for highlighting the best disinfection system based on reaction rate constant. The unique detailed investigation of P. aeruginosa disinfection with sunlight based disinfection systems under different weather conditions and variable parameters will help researchers to understand and further improve the newly invented SOCODIS system.

  2. Solar disinfection of Pseudomonas aeruginosa in harvested rainwater: a step towards potability of rainwater.

    Science.gov (United States)

    Amin, Muhammad T; Nawaz, Mohsin; Amin, Muhammad N; Han, Mooyoung

    2014-01-01

    Efficiency of solar based disinfection of Pseudomonas aeruginosa (P. aeruginosa) in rooftop harvested rainwater was evaluated aiming the potability of rainwater. The rainwater samples were exposed to direct sunlight for about 8-9 hours and the effects of water temperature (°C), sunlight irradiance (W/m2), different rear surfaces of polyethylene terephthalate bottles, variable microbial concentrations, pH and turbidity were observed on P. aeruginosa inactivation at different weathers. In simple solar disinfection (SODIS), the complete inactivation of P. aeruginosa was obtained only under sunny weather conditions (>50°C and >700 W/m2) with absorptive rear surface. Solar collector disinfection (SOCODIS) system, used to improve the efficiency of simple SODIS under mild and weak weather, completely inactivated the P. aeruginosa by enhancing the disinfection efficiency of about 20% only at mild weather. Both SODIS and SOCODIS systems, however, were found inefficient at weak weather. Different initial concentrations of P. aeruginosa and/or Escherichia coli had little effects on the disinfection efficiency except for the SODIS with highest initial concentrations. The inactivation of P. aeruginosa increased by about 10-15% by lowering the initial pH values from 10 to 3. A high initial turbidity, adjusted by adding kaolin, adversely affected the efficiency of both systems and a decrease, about 15-25%; in inactivation of P. aeruginosa was observed. The kinetics of this study was investigated by Geeraerd Model for highlighting the best disinfection system based on reaction rate constant. The unique detailed investigation of P. aeruginosa disinfection with sunlight based disinfection systems under different weather conditions and variable parameters will help researchers to understand and further improve the newly invented SOCODIS system.

  3. Insights into mechanisms and proteomic characterisation of Pseudomonas aeruginosa adaptation to a novel antimicrobial substance.

    Directory of Open Access Journals (Sweden)

    Peter Cierniak

    Full Text Available Antibiotic resistance has been reported since the introduction of synthetic antibiotics. Bacteria, such as one of the most common nosocomial pathogens P. aeruginosa, adapt quickly to changing environmental conditions, due to their short generation time. Thus microevolutional changes can be monitored in situ. In this study, the microevolutional process of Pseudomonas aeruginosa PAO1 resistance against a recently developed novel antibacterial zinc Schiff-base (ZSB was investigated at the proteome level. After extended exposure to ZSB the passaged strain differed in tolerance against ZSB, with the adapted P. aeruginosa PAO1 exhibiting 1.6 times higher minimal inhibitory concentration. Using Two-dimensional Difference Gel Electrophoresis, the changes in the proteome of ZSB adapted P. aeruginosa PAO1 were examined by comparison with the non-adapted P. aeruginosa PAO1. The proteome of the adapted P. aeruginosa PAO1 strain differed significantly from the non-adapted in the abundance of two proteins when both strains were grown under stressing conditions. One protein could be identified as the outer membrane protein D that plays a role in uptake of basic amino acids as well as in carbapeneme resistance. The second protein has been identified as alkyl peroxide reductase subunit F. Our data indicated a slight increase in abundance of alkyl peroxide reductase F (AhpF in the case of ZSB passaged P. aeruginosa PAO1. Higher abundance of Ahp has been discussed in the literature as a promoter of accelerated detoxification of benzene derivatives. The observed up-regulated AhpF thus appears to be connected to an increased tolerance against ZSB. Changes in the abundance of proteins connected to oxidative stress were also found after short-time exposure of P. aeruginosa PAO1 to the ZSB. Furthermore, adapted P. aeruginosa PAO1 showed increased tolerance against hydrogen peroxide and, in addition, showed accelerated degradation of ZSB, as determined by HPLC

  4. Hair removal in adolescence

    Directory of Open Access Journals (Sweden)

    Sandra Pereira

    2015-06-01

    Full Text Available Introduction: Due to hormonal stimulation during puberty, changes occur in hair type and distribution. In both sexes, body and facial unwanted hair may have a negative psychological impact on the teenager. There are several available methods of hair removal, but the choice of the most suitable one for each individual can raise doubts. Objective: To review the main methods of hair removal and clarify their indications, advantages and disadvantages. Development: There are several removal methods currently available. Shaving and depilation with chemicals products are temporary methods, that need frequent repetition, because hair removal is next to the cutaneous surface. The epilating methods in which there is full hair extraction include: epilation with wax, thread, tweezers, epilating machines, laser, intense pulsed light, and electrolysis. Conclusions: The age of beginning hair removal and the method choice must be individualized and take into consideration the skin and hair type, location, dermatological and endocrine problems, removal frequency, cost and personal preferences.

  5. Anti-Pseudomonas aeruginosa IgY antibodies augment bacterial clearance in a murine pneumonia model

    DEFF Research Database (Denmark)

    Thomsen, K.; Christophersen, L.; Bjarnsholt, T.

    2016-01-01

    -P. aeruginosa IgY antibodies on bacterial eradication in a murine pneumonia model. Methods: P. aeruginosa pneumonia was established in Balb/c mice and the effects of prophylactic IgY administration on lung bacteriology, clinical parameters and subsequent inflammation were compared to controls. Results......Background: Oral prophylactic therapy by gargling with pathogen-specific egg yolk immunoglobulins (IgY) may reduce the initial airway colonization with Pseudomonas aeruginosa in cystic fibrosis (CF) patients. IgY antibodies impart passive immunization and we investigated the effects of anti...

  6. Interference of Pseudomonas aeruginosa signalling and biofilm formation for infection control

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Tolker-Nielsen, Tim; Høiby, Niels;

    2010-01-01

    Pseudomonas aeruginosa is the best described bacterium with regards to quorum sensing (QS), in vitro biofilm formation and the development of antibiotic tolerance. Biofilms composed of P. aeruginosa are thought to be the underlying cause of many chronic infections, including those in wounds...... and in the lungs of patients with cystic fibrosis. In this review, we provide an overview of the molecular mechanisms involved in QS, QS-enabled virulence, biofilm formation and biofilm-enabled antibiotic tolerance. We now have substantial knowledge of the multicellular behaviour of P. aeruginosa in vitro. A major...

  7. Colistin-Tobramycin Combinations Are Superior to Monotherapy Concerning the Killing of Biofilm Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Herrmann, G.; Yang, Liang; Wu, H.

    2010-01-01

    biofilms. Methods. P. aeruginosa biofilms were generated in vitro and in rat lungs. In a pilot study, 5 patients with cystic fibrosis inhaled colistin and then tobramycin for 4 weeks. The changes in P. aeruginosa counts and lung function were assessed before and after therapy. Results. Antibiotic...... significantly lower after 7 days in animals receiving antibiotic combination than in animals receiving single antibiotics. In patients with cystic fibrosis, inhaled colistin-tobramycin was well tolerated and resulted in a mean decrease of 2.52 +/- 2.5 cfu of P. aeruginosa per milliliter of sputum (P = .027...

  8. Facultative control of matrix production optimizes competitive fitness in Pseudomonas aeruginosa PA14 biofilm models.

    Science.gov (United States)

    Madsen, Jonas S; Lin, Yu-Cheng; Squyres, Georgia R; Price-Whelan, Alexa; de Santiago Torio, Ana; Song, Angela; Cornell, William C; Sørensen, Søren J; Xavier, Joao B; Dietrich, Lars E P

    2015-12-01

    As biofilms grow, resident cells inevitably face the challenge of resource limitation. In the opportunistic pathogen Pseudomonas aeruginosa PA14, electron acceptor availability affects matrix production and, as a result, biofilm morphogenesis. The secreted matrix polysaccharide Pel is required for pellicle formation and for colony wrinkling, two activities that promote access to O2. We examined the exploitability and evolvability of Pel production at the air-liquid interface (during pellicle formation) and on solid surfaces (during colony formation). Although Pel contributes to the developmental response to electron acceptor limitation in both biofilm formation regimes, we found variation in the exploitability of its production and necessity for competitive fitness between the two systems. The wild type showed a competitive advantage against a non-Pel-producing mutant in pellicles but no advantage in colonies. Adaptation to the pellicle environment selected for mutants with a competitive advantage against the wild type in pellicles but also caused a severe disadvantage in colonies, even in wrinkled colony centers. Evolution in the colony center produced divergent phenotypes, while adaptation to the colony edge produced mutants with clear competitive advantages against the wild type in this O2-replete niche. In general, the structurally heterogeneous colony environment promoted more diversification than the more homogeneous pellicle. These results suggest that the role of Pel in community structure formation in response to electron acceptor limitation is unique to specific biofilm models and that the facultative control of Pel production is required for PA14 to maintain optimum benefit in different types of communities.

  9. Genetic Evidence for O-Specific Antigen as Receptor of Pseudomonas aeruginosa Phage K8 and Its Genomic Analysis

    Directory of Open Access Journals (Sweden)

    Xuewei ePan

    2016-03-01

    Full Text Available Phage therapy requires the comprehensive understanding of the mechanisms underlying the host-phage interactions. In this work, to identify the genes related to Pseudomonas aeruginosa phage K8 receptor synthesis, 16 phage-resistant mutants were selected from a Tn5G transposon mutant library of strain PAK. The disrupted genetic loci were identified and they were related to O-specific antigen (OSA synthesis, including gene wbpR, ssg, wbpV, wbpO, and Y880_RS05480, which encoded a putative O-antigen polymerase Wzy. The LPS profile of the Y880_RS05480 mutant was analyzed and shown to lack the O-antigen. Therefore, the data from characterization of Y880_RS05480 by TMHMM and SDS-PAGE silver staining analysis suggest that this locus might encode Wzy. The complete phage K8 genome was characterized as 93879 bp in length and contained identical 1188-bp terminal direct repeats. Comparative genomic analysis showed that phage K8 was highly homologous to members of the genus PaP1-like phages. On the basis of our genetic findings, OSA of P. aeruginosa PAK is proven to be the receptor of phage K8. The highly conserved structural proteins among the genetic closely related phages suggest that they may recognize the same receptor.

  10. Chronic lung infection by Pseudomonas aeruginosa biofilm is cured by L-Methionine in combination with antibiotic therapy.

    Science.gov (United States)

    Gnanadhas, Divya Prakash; Elango, Monalisha; Datey, Akshay; Chakravortty, Dipshikha

    2015-11-02

    Bacterial biofilms are associated with 80-90% of infections. Within the biofilm, bacteria are refractile to antibiotics, requiring concentrations >1,000 times the minimum inhibitory concentration. Proteins, carbohydrates and DNA are the major components of biofilm matrix. Pseudomonas aeruginosa (PA) biofilms, which are majorly associated with chronic lung infection, contain extracellular DNA (eDNA) as a major component. Herein, we report for the first time that L-Methionine (L-Met) at 0.5 μM inhibits Pseudomonas aeruginosa (PA) biofilm formation and disassembles established PA biofilm by inducing DNase expression. Four DNase genes (sbcB, endA, eddB and recJ) were highly up-regulated upon L-Met treatment along with increased DNase activity in the culture supernatant. Since eDNA plays a major role in establishing and maintaining the PA biofilm, DNase activity is effective in disrupting the biofilm. Upon treatment with L-Met, the otherwise recalcitrant PA biofilm now shows susceptibility to ciprofloxacin. This was reflected in vivo, in the murine chronic PA lung infection model. Mice treated with L-Met responded better to antibiotic treatment, leading to enhanced survival as compared to mice treated with ciprofloxacin alone. These results clearly demonstrate that L-Met can be used along with antibiotic as an effective therapeutic against chronic PA biofilm infection.

  11. ChIP-seq reveals the global regulator AlgR mediating cyclic di-GMP synthesis in Pseudomonas aeruginosa.

    Science.gov (United States)

    Kong, Weina; Zhao, Jingru; Kang, Huaping; Zhu, Miao; Zhou, Tianhong; Deng, Xin; Liang, Haihua

    2015-09-30

    AlgR is a key transcriptional regulator required for the expression of multiple virulence factors, including type IV pili and alginate in Pseudomonas aeruginosa. However, the regulon and molecular regulatory mechanism of AlgR have yet to be fully elucidated. Here, among 157 loci that were identified by a ChIP-seq assay, we characterized a gene, mucR, which encodes an enzyme that synthesizes the intracellular second messenger cyclic diguanylate (c-di-GMP). A ΔalgR strain produced lesser biofilm than did the wild-type strain, which is consistent with a phenotype controlled by c-di-GMP. AlgR positively regulates mucR via direct binding to its promoter. A ΔalgRΔmucR double mutant produced lesser biofilm than did the single ΔalgR mutant, demonstrating that c-di-GMP is a positive regulator of biofilm formation. AlgR controls the levels of c-di-GMP synthesis via direct regulation of mucR. In addition, the cognate sensor of AlgR, FimS/AlgZ, also plays an important role in P. aeruginosa virulence. Taken together, this study provides new insights into the AlgR regulon and reveals the involvement of c-di-GMP in the mechanism underlying AlgR regulation.

  12. Tobramycin Inhalation Powder™: a novel drug delivery system for treating chronic Pseudomonas aeruginosa infection in cystic fibrosis.

    Science.gov (United States)

    Parkins, Michael D; Elborn, J Stuart

    2011-10-01

    Lung disease in cystic fibrosis (CF) is typified by the development of chronic airways infection culminating in bronchiectasis and progression to end-stage respiratory disease. Pseudomonas aeruginosa, a ubiquitous gram-negative bacteria, is the archetypical CF pathogen and is associated with an accelerated clinical decline. The development and widespread use of chronic suppressive aerosolized antibacterial therapies, in particular Tobramycin Inhalation Solution (TIS), in CF has contributed to reduced lung function decline and improved survival. However, the requirement for the aerosolization of these agents through nebulizers has been associated with increased treatment burden, reduced quality of life and remain a barrier to broader uptake. Tobramycin Inhalation Powder (TIP™) has been developed by Novartis with the express purpose of delivering the same benefits as TIS in a time-effective manner. Administered via the T-326™ (Novartis) Inhaler in four individual 28-mg capsules, TIP can be administered in a quarter of the time of traditional nebulizers and is inherently portable. In clinical studies, TIP has been shown to be safe, result in equivalent or superior reductions in P. aeruginosa sputum density and produce similar improvements in pulmonary function. TIP offers significant advantages in time saving, portability and convenience over traditional nebulized TIS with comparable clinical outcomes for individuals with CF.

  13. Assessment of Pseudomonas aeruginosa N5,N10-methylenetetrahydrofolate dehydrogenase-cyclohydrolase as a potential antibacterial drug target.

    Directory of Open Access Journals (Sweden)

    Thomas C Eadsforth

    Full Text Available The bifunctional enzyme methylenetetrahydrofolate dehydrogenase - cyclohydrolase (FolD is identified as a potential drug target in Gram-negative bacteria, in particular the troublesome Pseudomonas aeruginosa. In order to provide a comprehensive and realistic assessment of the potential of this target for drug discovery we generated a highly efficient recombinant protein production system and purification protocol, characterized the enzyme, carried out screening of two commercial compound libraries by differential scanning fluorimetry, developed a high-throughput enzyme assay and prosecuted a screening campaign against almost 80,000 compounds. The crystal structure of P. aeruginosa FolD was determined at 2.2 Å resolution and provided a template for an assessment of druggability and for modelling of ligand complexes as well as for comparisons with the human enzyme. New FolD inhibitors were identified and characterized but the weak levels of enzyme inhibition suggest that these compounds are not optimal starting points for future development. Furthermore, the close similarity of the bacterial and human enzyme structures suggest that selective inhibition might be difficult to attain. In conclusion, although the preliminary biological data indicates that FolD represents a valuable target for the development of new antibacterial drugs, indeed spurred us to investigate it, our screening results and structural data suggest that this would be a difficult enzyme to target with respect to developing the appropriate lead molecules required to underpin a serious drug discovery effort.

  14. Effect of pH on biologic degradation of Microcystis aeruginosa by alga-lysing bacteria in sequencing batch biofilm reactors

    Institute of Scientific and Technical Information of China (English)

    Hongjing LI; Mengli HAO; Jingxian LIU; Chen CHEN1; Zhengqiu FAN; Xiangrong WANG

    2012-01-01

    In this paper, the effect of pH on biological degradation of Microcystis aeruginosa by alga-lysing bacteria in laboratory-scale sequencing batch biofilm reactors (SBBRs) was investigated. After 10 d filming with waste activated sludge, the biological film could be formed, and the bioreactors in which laid polyolefin resin filler were used to treat algal culture. By comparing the removal efficiency of chlorophyll a at different aerobic time, the optimum time was determined as 5 h. Under pH 6.5, 7.5, and 8.5 conditions, the removal rates of Microcystis aeruginosa were respectively 75.9%, 83.6%, and 78.3% (in term of chlorophyll a), and that of Chemical Oxygen Demand (CODMn) were 30.6%, 35.8%, and 33.5%. While the removal efficiencies of ammonia nitrogen (NH+ -N) were all 100%. It was observed that the sequence of the removal efficiencies of algae, NH+ -N and organic matter were pH 7.5 〉 pH 8.5 〉 pH 6.5. The results showed that the dominant alga-lysing bacteria in the SBBRs was strain HM-01, which was identified as Bacillus sp. by Polymerase Chain Reaction (PCR) amplification of the 16S rRNA gene, Basic Local Alignment Search Tool (BLAST) analysis, and compar- ison with sequences in the GenBank nucleotide database. The algicidal activated substance which HM-01 strain excreted could withstand high temperature and pressure, also had better hydrophily and stronger polarity.

  15. Particle adhesion and removal

    CERN Document Server

    Mittal, K L

    2015-01-01

    The book provides a comprehensive and easily accessible reference source covering all important aspects of particle adhesion and removal.  The core objective is to cover both fundamental and applied aspects of particle adhesion and removal with emphasis on recent developments.  Among the topics to be covered include: 1. Fundamentals of surface forces in particle adhesion and removal.2. Mechanisms of particle adhesion and removal.3. Experimental methods (e.g. AFM, SFA,SFM,IFM, etc.) to understand  particle-particle and particle-substrate interactions.4. Mechanics of adhesion of micro- and  n

  16. Region 9 Removal Sites

    Data.gov (United States)

    U.S. Environmental Protection Agency — Point geospatial dataset representing locations of CERCLA (Superfund) Removal sites. CERCLA (Comprehensive Environmental Response, Compensation, and Liability Act)...

  17. Adhesion of Pseudomonas aeruginosa strains to untreated and oxygen-plasma treated poly(vinyl chloride) (PVC) from endotracheal intubation devices.

    Science.gov (United States)

    Triandafillu, K; Balazs, D J; Aronsson, B-O; Descouts, P; Tu Quoc, P; van Delden, C; Mathieu, H J; Harms, H

    2003-04-01

    Pseudomonas aeruginosa pneumonia is a life threatening complication in mechanically ventilated patients that requires the ability of the bacteria to adhere to, and colonize the endotracheal intubation device. New strategies to prevent or reduce these nosocomial infections are greatly needed. We report here the study of a set of P. aeruginosa clinical isolates, together with specific mutants, regarding their adhesion on native and chemically modified poly(vinyl chloride) (PVC) surfaces from endotracheal intubation devices. The adhesion of the different strains to untreated PVC varied widely, correlating with several physico-chemical characteristics known to influence the attachment of bacteria to inert surfaces. The adhesion patterns were compared to the calculations obtained with the DLVO theory of colloidal stability. These results illustrate the importance of testing different clinical isolates when investigating bacterial adhesion. Oxygen plasma treatment of the PVC pieces yielded a hydrophilic surface and reduced the number of adhering bacteria by as much as 70%. This reduction is however unlikely to be sufficient to prevent P. aeruginosa colonization of endotracheal intubation devices.

  18. Co-Carriage of blaKPC-2 and blaNDM-1 in Clinical Isolates of Pseudomonas aeruginosa Associated with Hospital Infections from India.

    Directory of Open Access Journals (Sweden)

    Deepjyoti Paul

    Full Text Available Global spread of KPC poses to be a serious threat complicating treatment options in hospital settings. The present study investigates the genetic environment of blaKPC-2 among clinical isolates of Pseudomonas aeruginosa from a tertiary referral hospital of India. The study isolates were collected from different wards and clinics of Silchar Medical College and Hospital, India, from 2012-2013. The presence of blaKPC was confirmed by genotypic characterization followed by sequencing. Cloning of the blaKPC-2 gene was performed and the genetic environment of this gene was characterized as well. Transferability of the resistance gene was determined by transformation assay and Southern hybridization. Additionally, restriction mapping was also carried out. Two isolates of P. aeruginosa were found to harbor blaKPC-2, were resistant towards aminoglycosides, quinolone and β-lactam-β-lactamase inhibitor combination. In both the isolates, the resistance determinant was associated with class 1 integron and horizontally transferable. Both the isolates were co-harboring blaNDM-1. The first detection of this integron mediated blaKPC-2 coexisting with blaNDM-1 in P. aeruginosa from India is worrisome, and further investigation is required to track the gene cassette mediated blaKPC-2 in terms of infection control and to prevent the spread of this gene in hospitals as well as in the community.

  19. A novel Pseudomonas aeruginosa bacteriophage, Ab31, a chimera formed from temperate phage PAJU2 and P. putida lytic phage AF: characteristics and mechanism of bacterial resistance.

    Directory of Open Access Journals (Sweden)

    Libera Latino

    Full Text Available A novel temperate bacteriophage of Pseudomonas aeruginosa, phage vB_PaeP_Tr60_Ab31 (alias Ab31 is described. Its genome is composed of structural genes related to those of lytic P. putida phage AF, and regulatory genes similar to those of temperate phage PAJU2. The virion structure resembles that of phage AF and other lytic Podoviridae (S. enterica Epsilon 15 and E. coli phiv10 with similar tail spikes. Ab31 was able to infect P. aeruginosa strain PA14 and two genetically related strains called Tr60 and Tr162, out of 35 diverse strains from cystic fibrosis patients. Analysis of resistant host variants revealed different phenotypes, including induction of pigment and alginate overproduction. Whole genome sequencing of resistant variants highlighted the existence of a large deletion of 234 kbp in two strains, encompassing a cluster of genes required for the production of CupA fimbriae. Stable lysogens formed by Ab31 in strain Tr60, permitted the identification of the insertion site. During colonization of the lung in cystic fibrosis patients, P. aeruginosa adapts by modifying its genome. We suggest that bacteriophages such as Ab31 may play an important role in this adaptation by selecting for bacterial characteristics that favor persistence of bacteria in the lung.

  20. Early activation of quorum sensing in Pseudomonas aeruginosa reveals the architecture of a complex regulon

    Directory of Open Access Journals (Sweden)

    Schuster Martin

    2007-08-01

    Full Text Available Abstract Background Quorum-sensing regulation of gene expression in Pseudomonas aeruginosa is complex. Two interconnected acyl-homoserine lactone (acyl-HSL signal-receptor pairs, 3-oxo-dodecanoyl-HSL-LasR and butanoyl-HSL-RhlR, regulate more than 300 genes. The induction of most of the genes is delayed during growth of P. aeruginosa in complex medium, cannot be advanced by addition of exogenous signal, and requires additional regulatory components. Many of these late genes can be induced by addition of signals early by using specific media conditions. While several factors super-regulate the quorum receptors, others may co-regulate target promoters or may affect expression posttranscriptionally. Results To better understand the contributions of super-regulation and co-regulation to quorum-sensing gene expression, and to better understand the general structure of the quorum sensing network, we ectopically expressed the two receptors (in the presence of their cognate signals and another component that affects quorum sensing, the stationary phase sigma factor RpoS, early in growth. We determined the effect on target gene expression by microarray and real-time PCR analysis. Our results show that many target genes (e.g. lasB and hcnABC are directly responsive to receptor protein levels. Most genes (e.g. lasA, lecA, and phnAB, however, are not significantly affected, although at least some of these genes are directly regulated by quorum sensing. The majority of promoters advanced by RhlR appeared to be regulated directly, which allowed us to build a RhlR consensus sequence. Conclusion The direct responsiveness of many quorum sensing target genes to receptor protein levels early in growth confirms the role of super-regulation in quorum sensing gene expression. The observation that the induction of most target genes is not affected by signal or receptor protein levels indicates that either target promoters are co-regulated by other transcription factors

  1. Production of biopolymers by Pseudomonas aeruginosa isolated from marine source

    Directory of Open Access Journals (Sweden)

    Nazia Jamil

    2008-06-01

    Full Text Available Two bacterial strains, Pseudomonas aeruginosa CMG607w and CMG1421 produce commercially important biopolymers. CMG607w isolated from the sediments of Lyari outfall to Arabian Sea synthesize the mcl-polyhydroxyalkanoates from various carbon sources. The production of PHAs was directly proportional to the incubation periods. Other strain CMG1421, a dry soil isolate, produced high viscous water absorbing extracellular acidic polysaccharide when it was grown aerobically in the minimal medium containing glucose or fructose or sucrose as sole source of carbon. The biopolymer had the ability to absorb water 400 times more than its dry weight. This property was superior to that of currently used non-degradable synthetic water absorbents. It acted as salt filter and had rheological and stabilizing activity as well.

  2. Attenuation of Pseudomonas aeruginosa virulence by quorum sensing inhibitors

    DEFF Research Database (Denmark)

    Hentzer, Morten; Wu, H.; Andersen, Jens Bo;

    2003-01-01

    Traditional treatment of infectious diseases is based on compounds that kill or inhibit growth of bacteria. A major concern with this approach is the frequent development of resistance to antibiotics. The discovery of communication systems (quorum sensing systems) regulating bacterial virulence has...... of natural furanone compounds can act as a potent antagonist of bacterial quorum sensing. We employed GeneChip((R)) microarray technology to identify furanone target genes and to map the quorum sensing regulon. The transcriptome analysis showed that the furanone drug specifically targeted quorum sensing...... systems and inhibited virulence factor expression. Application of the drug to P.aeruginosa biofilms increased bacterial susceptibility to tobramycin and SDS. In a mouse pulmonary infection model, the drug inhibited quorum sensing of the infecting bacteria and promoted their clearance by the mouse immune...

  3. Antibacterial Coating for Elimination of Pseudomonas aeruginosa and Escherichia coli

    Directory of Open Access Journals (Sweden)

    Zainal Abidin Ali

    2014-01-01

    Full Text Available A polymer antibacterial surface has been successfully developed. The coating system used silane as binder and Ag particles as antibacterial agent. The silver was synthesized using precipitation method. X-ray diffraction (XRD, field emission scanning electron microscopy (FESEM, Brunauer-Emmett-Teller (BET tests, energy-dispersive X-ray spectroscopy (EDX, and X-ray photoelectron spectroscopy (XPS were carried out to evaluate the silver particles. Antibacterial properties of the coating system were tested against gram-negative bacteria, namely, Pseudomonas aeruginosa and Escherichia coli. Different amounts of Ag were used in the coating to optimize its usage. The Japanese International Standard, JISZ2801, was used for bacteria test and the surface developed complies with the standard being antibacterial.

  4. Decrease of Pseudomonas aeruginosa biofilm formation by food waste materials.

    Science.gov (United States)

    Maderova, Zdenka; Horska, Katerina; Kim, Sang-Ryoung; Lee, Chung-Hak; Pospiskova, Kristyna; Safarikova, Mirka; Safarik, Ivo

    2016-01-01

    The formation of bacterial biofilm on various surfaces has significant negative economic effects. The aim of this study was to find a simple procedure to decrease the Pseudomonas aeruginosa biofilm formation in a water environment by using different food waste biological materials as signal molecule adsorbents. The selected biomaterials did not reduce the cell growth but affected biofilm formation. Promising biomaterials were magnetically modified in order to simplify manipulation and facilitate their magnetic separation. The best biocomposite, magnetically modified spent grain, exhibited substantial adsorption of signal molecules and decreased the biofilm formation. These results suggest that selected food waste materials and their magnetically responsive derivatives could be applied to solve biofilm problems in water environment.

  5. Novel multiscale modeling tool applied to Pseudomonas aeruginosa biofilm formation.

    Science.gov (United States)

    Biggs, Matthew B; Papin, Jason A

    2013-01-01

    Multiscale modeling is used to represent biological systems with increasing frequency and success. Multiscale models are often hybrids of different modeling frameworks and programming languages. We present the MATLAB-NetLogo extension (MatNet) as a novel tool for multiscale modeling. We demonstrate the utility of the tool with a multiscale model of Pseudomonas aeruginosa biofilm formation that incorporates both an agent-based model (ABM) and constraint-based metabolic modeling. The hybrid model correctly recapitulates oxygen-limited biofilm metabolic activity and predicts increased growth rate via anaerobic respiration with the addition of nitrate to the growth media. In addition, a genome-wide survey of metabolic mutants and biofilm formation exemplifies the powerful analyses that are enabled by this computational modeling tool.

  6. Novel multiscale modeling tool applied to Pseudomonas aeruginosa biofilm formation.

    Directory of Open Access Journals (Sweden)

    Matthew B Biggs

    Full Text Available Multiscale modeling is used to represent biological systems with increasing frequency and success. Multiscale models are often hybrids of different modeling frameworks and programming languages. We present the MATLAB-NetLogo extension (MatNet as a novel tool for multiscale modeling. We demonstrate the utility of the tool with a multiscale model of Pseudomonas aeruginosa biofilm formation that incorporates both an agent-based model (ABM and constraint-based metabolic modeling. The hybrid model correctly recapitulates oxygen-limited biofilm metabolic activity and predicts increased growth rate via anaerobic respiration with the addition of nitrate to the growth media. In addition, a genome-wide survey of metabolic mutants and biofilm formation exemplifies the powerful analyses that are enabled by this computational modeling tool.

  7. Identification of microcystins from three collection strains of Microcystis aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Campo, Francisca F. del [Departamento de Biologia, Universidad Autonoma de Madrid, Cantoblanco, 28049-Madrid (Spain); Ouahid, Youness, E-mail: ouahidyouness@gmail.co [Departamento de Biologia, Universidad Autonoma de Madrid, Cantoblanco, 28049-Madrid (Spain)

    2010-09-15

    Microcystins (MCs) are toxic cyclic heptapeptides produced by various cyanobacteria genera, especially Microcystis. We identified 10 out of 12 MCs produced by three Microcystis aeruginosa strains from cyanobacteria collections, UTEX 2666, UTEX 2670 and UAM 1303, by using two analytical methods: Matrix-assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF/MS) and HPLC Photodiode Array Detector coupled to a hybrid Quadrupole Time of Flight Mass Spectrometry (HPLC-PDA-QTOF/MS). MALDI-TOF/MS failed to detect non-polar MCs, such as MC-LY and MC-LW. HPLC-QTOF/MS permitted the accurate identification of most MCs present in methanolic extracts. Besides, three new MCs, namely: [D-Glu(OCH{sub 3}){sup 6}, D-Asp{sup 3}] MC-LAba, MC-YL and MC-YM were detected by HPLC-QTOF/MS. - Three new microcystin variants identified by HPLC-QTOF/MS.

  8. Evolution and Pathoadaptation of Pseudomonas aeruginosa in Cystic Fibrosis Patients

    DEFF Research Database (Denmark)

    Marvig, Rasmus Lykke

    . While this phenotype may accelerate evolution, we also showed that hypermutators display differential mutagenesis of certain genes which enable them to follow alternative evolutionary pathways. Overall, our study identifies genes important for bacterial adaptation to a human host environment......Molecular and mechanistic understanding of evolution is essential for our ability to comprehend the development of life on Earth. Life appeared around 4 billion years ago, and has ever since adapted and diversified through the process of evolution. The focus of this thesis has been to increase our...... understanding of how bacteria evolve and genetically adapt in a natural environment. In particular we sought to identify the genes that are targeted by mutation to optimize fitness in a given environment, and to understand the evolutionary mechanisms that govern the genetic change. Pseudomonas aeruginosa...

  9. Magnetic fields suppress Pseudomonas aeruginosa biofilms and enhance ciprofloxacin activity.

    Science.gov (United States)

    Bandara, H M H N; Nguyen, D; Mogarala, S; Osiñski, M; Smyth, H D C

    2015-01-01

    Due to the refractory nature of pathogenic microbial biofilms, innovative biofilm eradication strategies are constantly being sought. Thus, this study addresses a novel approach to eradicate Pseudomonas aeruginosa biofilms. Magnetic nanoparticles (MNP), ciprofloxacin (Cipro), and magnetic fields were systematically evaluated in vitro for their relative anti-biofilm contributions. Twenty-four-hour biofilms exposed to aerosolized MNPs, Cipro, or a combination of both, were assessed in the presence or absence of magnetic fields (Static one-sided, Static switched, Oscillating, Static + oscillating) using changes in bacterial metabolism, biofilm biomass, and biofilm imaging. The biofilms exposed to magnetic fields alone exhibited significant metabolic and biomass reductions (p biofilms were treated with a MNP/Cipro combination, the most significant metabolic and biomass reductions were observed when exposed to static switched magnetic fields (p biofilms to a static switched magnetic field alone, or co-administration with MNP/Cipro/MNP + Cipro appears to be a promising approach to eradicate biofilms of this bacterium.

  10. The implication of Pseudomonas aeruginosa biofilms in infections

    DEFF Research Database (Denmark)

    Rybtke, Morten Theil; Jensen, Peter Ø; Høiby, Niels

    2011-01-01

    of infection in the lungs of cystic fibrosis patients and in chronic wounds. In this review we address the molecular basis of biofilm development by P. aeruginosa as well as the mechanisms employed by this bacterium in the increased tolerance displayed against antimicrobials. The complex build......-up of the extracellular matrix encasing the biofilm-associated bacteria as well as the elaborate signaling mechanisms employed by the bacterium enables it to withstand the continuous stresses imposed by the immune defense and administered antibiotics resulting in a state of chronic inflammation that damages the host...... treatment strategies where the underlying targets are less prone for resistance development as bacteria, in retrospect, have a unique ability to evade the actions of classic antibiotics....

  11. Experimental Pseudomonas aeruginosa mediated rhino sinusitis in mink

    DEFF Research Database (Denmark)

    Kirkeby, S.; Hammer, A. S.; Høiby, N.

    2017-01-01

    The nasal and sinus cavities in children may serve as reservoirs for microorganisms that cause recurrent and chronic lung infections. This study evaluates whether the mink can be used as an animal model for studying Pseudomonas aeruginosa mediated rhino-sinusitis since there is no suitable...... traditional animal model for this disease. Nasal tissue samples from infected and control mink were fixed in formalin, demineralized, and embedded in paraffin. A histological examination of sections from the infected animals revealed disintegration of the respiratory epithelium lining the nasal turbinates...... and swelling and edema of the submucosa. The expression of mucins and sialylated glycans was examined using immunohistochemistry. MUC1, MUC2 and MUC5AC were upregulated in the inoculated animals as a much stronger staining was present in the respiratory epithelium in the infected animals compared...

  12. Bioengineered lysozyme in combination therapies for Pseudomonas aeruginosa lung infections

    Science.gov (United States)

    Griswold, Karl E; Bement, Jenna L; Teneback, Charlotte C; Scanlon, Thomas C; Wargo, Matthew J; Leclair, Laurie W

    2014-01-01

    There is increasing urgency in the battle against drug-resistant bacterial pathogens, and this public health crisis has created a desperate need for novel antimicrobial agents. Recombinant human lysozyme represents one interesting candidate for treating pulmonary infections, but the wild type enzyme is subject to electrostatic mediated inhibition by anionic biopolymers that accumulate in the infected lung. We have redesigned lysozyme’s electrostatic potential field, creating a genetically engineered variant that is less susceptible to polyanion inhibition, yet retains potent bactericidal activity. A recent publication demonstrated that the engineered enzyme outperforms wild type lysozyme in a murine model of Pseudomonas aeruginosa lung infection. Here, we expand upon our initial studies and consider dual therapies that combine lysozymes with an antimicrobial peptide. Consistent with our earlier results, the charge modified lysozyme combination outperformed its wild type counterpart, yielding more than an order-of-magnitude reduction in bacterial burden following treatment with a single dose. PMID:24637705

  13. Medium factors on anaerobic production of rhamnolipids by Pseudomonas aeruginosa SG and a simplifying medium for in situ microbial enhanced oil recovery applications.

    Science.gov (United States)

    Zhao, Feng; Zhou, Jidong; Han, Siqin; Ma, Fang; Zhang, Ying; Zhang, Jie

    2016-04-01

    Aerobic production of rhamnolipid by Pseudomonas aeruginosa was extensively studied. But effect of medium composition on anaerobic production of rhamnolipid by P. aeruginosa was unknown. A simplifying medium facilitating anaerobic production of rhamnolipid is urgently needed for in situ microbial enhanced oil recovery (MEOR). Medium factors affecting anaerobic production of rhamnolipid were investigated using P. aeruginosa SG (Genbank accession number KJ995745). Medium composition for anaerobic production of rhamnolipid by P. aeruginosa is different from that for aerobic production of rhamnolipid. Both hydrophobic substrate and organic nitrogen inhibited rhamnolipid production under anaerobic conditions. Glycerol and nitrate were the best carbon and nitrogen source. The commonly used N limitation under aerobic conditions was not conducive to rhamnolipid production under anaerobic conditions because the initial cell growth demanded enough nitrate for anaerobic respiration. But rhamnolipid was also fast accumulated under nitrogen starvation conditions. Sufficient phosphate was needed for anaerobic production of rhamnolipid. SO4(2-) and Mg(2+) are required for anaerobic production of rhamnolipid. Results will contribute to isolation bacteria strains which can anaerobically produce rhamnolipid and medium optimization for anaerobic production of rhamnolipid. Based on medium optimization by response surface methodology and ions composition of reservoir formation water, a simplifying medium containing 70.3 g/l glycerol, 5.25 g/l NaNO3, 5.49 g/l KH2PO4, 6.9 g/l K2HPO4·3H2O and 0.40 g/l MgSO4 was designed. Using the simplifying medium, 630 mg/l of rhamnolipid was produced by SG, and the anaerobic culture emulsified crude oil to EI24 = 82.5 %. The simplifying medium was promising for in situ MEOR applications.

  14. Degradation characteristics of two Bacillus strains on the Microcystis aeruginosa

    Institute of Scientific and Technical Information of China (English)

    PEI Hai-yan; HU Wen-rong; QU Yin-bo; MU Rui-min; LI Xiao-cai

    2005-01-01

    The degradation kinetics of strains P05 and P07 and the degradation effects of mixed strain on Microcystis aeruginosa were studied. The results showed that: ( 1 ) The degradation processes of strains P05 and P07 on Microcystis aeruginosa accorded with the first-order reaction model when the range of Chl- a concentration was from 0 to 1500 μg/L. (2) The initial bacterium densities had a strong influence on the degradation velocity. The greater the initial bacterium density was, the faster the degradation was. The degradation velocity constants of P05 were 0.1913, 0.2175 and 0.3092 respectively, when bacterium densities were 4.8×105 , 4.8 × 106, 2.4 × 107 cells/ml. For strain P07, they were 0.1509, 0.1647 and 0.2708. The degradation velocity constant of strain P05 was higher than that of P07 when the bacterium density was under 4.8 × 105 cells/ml, but the constant increasing of P07 was quicker than that of P05. (3) The degradation effects of P05 and P07 strains did not antagonize. When the concentration of Chl-a was high, the degradation effects of mixed strain excelled that of any single strains. But with the decrease of the Chl-a concentration, this advantage was not clear. When the concentration was less than 180 μg/L, the degradation effects of mixed were consistent with that of strain P07.

  15. Fructooligosacharides reduce Pseudomonas aeruginosa PAO1 pathogenicity through distinct mechanisms.

    Directory of Open Access Journals (Sweden)

    Mercedes Ortega-González

    Full Text Available Pseudomonas aeruginosa is ubiquitously present in the environment and acts as an opportunistic pathogen on humans, animals and plants. We report here the effects of the prebiotic polysaccharide inulin and its hydrolysed form FOS on this bacterium. FOS was found to inhibit bacterial growth of strain PAO1, while inulin did not affect growth rate or yield in a significant manner. Inulin stimulated biofilm formation, whereas a dramatic reduction of the biofilm formation was observed in the presence of FOS. Similar opposing effects were observed for bacterial motility, where FOS inhibited the swarming and twitching behaviour whereas inulin caused its stimulation. In co-cultures with eukaryotic cells (macrophages FOS and, to a lesser extent, inulin reduced the secretion of the inflammatory cytokines IL-6, IL-10 and TNF-α. Western blot experiments indicated that the effects mediated by FOS in macrophages are associated with a decreased activation of the NF-κB pathway. Since FOS and inulin stimulate pathway activation in the absence of bacteria, the FOS mediated effect is likely to be of indirect nature, such as via a reduction of bacterial virulence. Further, this modulatory effect is observed also with the highly virulent ptxS mutated strain. Co-culture experiments of P. aeruginosa with IEC18 eukaryotic cells showed that FOS reduces the concentration of the major virulence factor, exotoxin A, suggesting that this is a possible mechanism for the reduction of pathogenicity. The potential of these compounds as components of antibacterial and anti-inflammatory cocktails is discussed.

  16. Phosphorylcholine Phosphatase: A Peculiar Enzyme of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Domenech

    2011-01-01

    Full Text Available Pseudomonas aeruginosa synthesizes phosphorylcholine phosphatase (PchP when grown on choline, betaine, dimethylglycine or carnitine. In the presence of Mg2+ or Zn2+, PchP catalyzes the hydrolysis of p-nitrophenylphosphate (p-NPP or phosphorylcholine (Pcho. The regulation of pchP gene expression is under the control of GbdR and NtrC; dimethylglycine is likely the metabolite directly involved in the induction of PchP. Therefore, the regulation of choline metabolism and consequently PchP synthesis may reflect an adaptive response of P. aeruginosa to environmental conditions. Bioinformatic and biochemistry studies shown that PchP contains two sites for alkylammonium compounds (AACs: one in the catalytic site near the metal ion-phosphoester pocket, and another in an inhibitory site responsible for the binding of the alkylammonium moiety. Both sites could be close to each other and interact through the residues 42E, 43E and 82YYY84. Zn2+ is better activator than Mg2+ at pH 5.0 and it is more effective at alleviating the inhibition produced by the entry of Pcho or different AACs in the inhibitory site. We postulate that Zn2+ induces at pH 5.0 a conformational change in the active center that is communicated to the inhibitory site, producing a compact or closed structure. However, at pH 7.4, this effect is not observed because to the hydrolysis of the [Zn2+L2−1L20(H2O2] complex, which causes a change from octahedral to tetrahedral in the metal coordination geometry. This enzyme is also present in P. fluorescens, P. putida, P. syringae, and other organisms. We have recently crystallized PchP and solved its structure.

  17. Structural Characterization of Novel Pseudomonas aeruginosa Type IV Pilins

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Y.; Jackson, S; Aidoo, F; Junop, M; Burrows, L

    2010-01-01

    Pseudomonas aeruginosa type IV pili, composed of PilA subunits, are used for attachment and twitching motility on surfaces. P. aeruginosa strains express one of five phylogenetically distinct PilA proteins, four of which are associated with accessory proteins that are involved either in pilin posttranslational modification or in modulation of pilus retraction dynamics. Full understanding of pilin diversity is crucial for the development of a broadly protective pilus-based vaccine. Here, we report the 1.6-{angstrom} X-ray crystal structure of an N-terminally truncated form of the novel PilA from strain Pa110594 (group V), which represents the first non-group II pilin structure solved. Although it maintains the typical T4a pilin fold, with a long N-terminal {alpha}-helix and four-stranded antiparallel {beta}-sheet connected to the C-terminus by a disulfide-bonded loop, the presence of an extra helix in the {alpha}{beta}-loop and a disulfide-bonded loop with helical character gives the structure T4b pilin characteristics. Despite the presence of T4b features, the structure of PilA from strain Pa110594 is most similar to the Neisseria gonorrhoeae pilin and is also predicted to assemble into a fiber similar to the GC pilus, based on our comparative pilus modeling. Interactions between surface-exposed areas of the pilin are suggested to contribute to pilus fiber stability. The non-synonymous sequence changes between group III and V pilins are clustered in the same surface-exposed areas, possibly having an effect on accessory protein interactions. However, based on our high-confidence model of group III PilA{sub PA14}, compensatory changes allow for maintenance of a similar shape.

  18. Lichen secondary metabolite evernic acid as potential quorum sensing inhibitor against Pseudomonas aeruginosa.

    Science.gov (United States)

    Gökalsın, Barış; Sesal, Nüzhet Cenk

    2016-09-01

    Cystic Fibrosis is a genetic disease and it affects the respiratory and digestive systems. Pseudomonas aeruginosa infections in Cystic Fibrosis are presented as the main cause for high mortality and morbidity rates. Pseudomonas aeruginosa populations can regulate their virulence gene expressions via the bacterial communication system: quorum sensing. Inhibition of quorum sensing by employing quorum sensing inhibitors can leave the bacteria vulnerable. Therefore, determining natural sources to obtain potential quorum sensing inhibitors is essential. Lichens have ethnobotanical value for their medicinal properties and it is possible that their secondary metabolites have quorum sensing inhibitor properties. This study aims to investigate an alternative treatment approach by utilizing lichen secondary metabolite evernic acid to reduce the expressions of Pseudomonas aeruginosa virulence factors by inhibiting quorum sensing. For this purpose, fluorescent monitor strains were utilized for quorum sensing inhibitor screens and quantitative reverse-transcriptase PCR analyses were conducted for comparison. Results indicate that evernic acid is capable of inhibiting Pseudomonas aeruginosa quorum sensing systems.

  19. Flavonoids from Rhizophora conjugata fruit extract blocks virulence factors of Pseudomonas aeruginosa

    Digital Repository Service at National Institute of Oceanography (India)

    Naik, D.; Tilvi, S.; DeSouza, L.

    : chloroform (1:1) extracts of 7 mangrove plants on P. aeruginosa ATCC 27853 motilities (swarming, swimming and twitching). Amongst the 22 extracts tested, methanolic extract of Rhizophora conjugata fruit showed maximum inhibition. The butanol (RcBu) fraction...

  20. Respiratory syncytial virus infection facilitates acute colonization of Pseudomonas aeruginosa in mice

    DEFF Research Database (Denmark)

    de Vrankrijker, Angélica M M; Wolfs, Tom F W; Ciofu, Oana;

    2009-01-01

    Pseudomonas aeruginosa causes opportunistic infections in immunocompromised individuals and patients ventilated mechanically and is the major pathogen in patients with cystic fibrosis, in which it causes chronic infections. Epidemiological, in vitro and animal data suggest a role for respiratory ...

  1. Activation of human pro-urokinase by unrelated proteases secreted by Pseudomonas aeruginosa.

    NARCIS (Netherlands)

    Beaufort, N.; Seweryn, P.; Bentzmann, S. de; Tang, A.; Kellermann, J.; Grebenchtchikov, N.J.; Schmitt, M.; Sommerhoff, C.P.; Pidard, D.; Magdolen, V.

    2010-01-01

    Pathogenic bacteria, including Pseudomonas aeruginosa, interact with and engage the host plasminogen (Plg) activation system, which encompasses the urokinase (uPA)-type Plg activator, and is involved in extracellular proteolysis, including matrilysis and fibrinolysis. We hypothesized that secreted b

  2. Pseudo-outbreak of pseudomonas aeruginosa in HIV-infected patients undergoing fiberoptic bronchoscopy

    DEFF Research Database (Denmark)

    Kolmos, H J; Lerche, A; Kristoffersen, Kirsten Lydia;

    1994-01-01

    Pseudomonas aeruginosa was isolated from bronchoalveolar lavage fluid from 8 consecutive patients undergoing bronchoscopy at an infectious diseases unit. None of the patients developed signs of respiratory tract infection that could be ascribed to the organism. The source of contamination...

  3. Phage-antibiotic synergism: a possible approach to combatting Pseudomonas aeruginosa.

    Science.gov (United States)

    Knezevic, Petar; Curcin, Sanja; Aleksic, Verica; Petrusic, Milivoje; Vlaski, Ljiljana

    2013-01-01

    Pseudomonas aeruginosa is a highly resistant opportunistic pathogen and an important etiological agent of various types of infections. During the last decade, P. aeruginosa phages have been extensively examined as alternative antimicrobial agents. The aim of the study was to determine antimicrobial effectiveness of combining subinhibitory concentrations of gentamicin, ceftriaxone, ciprofloxacin or polymyxin B with P. aeruginosa-specific bacteriophages belonging to families Podoviridae and Siphoviridae. The time-kill curve method showed that a combination of bacteriophages and subinhibitory concentrations of ceftriaxone generally reduced bacterial growth, and synergism was proven for a Siphoviridae phage σ-1 after 300 min of incubation. The detected alteration in morphology after ceftriaxone application, resulting in cell elongation, along with its specific mode of action, seemed to be a necessary but was not a sufficient reason for phage-antibiotic synergism. The phenomenon offers an opportunity for future development of treatment strategies for potentially lethal infections caused by P. aeruginosa.

  4. Within-host evolution of Pseudomonas aeruginosa reveals adaptation toward iron acquisition from hemoglobin

    DEFF Research Database (Denmark)

    Marvig, Rasmus Lykke; Pedersen, Søren Damkiær; Khademi, Seyed Mohammad Hossein;

    2014-01-01

    Pseudomonas aeruginosa airway infections are a major cause of mortality and morbidity of cystic fibrosis (CF) patients. In order to persist, P. aeruginosa depends on acquiring iron from its host, and multiple different iron acquisition systems may be active during infection. This includes...... the pyoverdine siderophore and the Pseudomonas heme utilization (phu) system. While the regulation and mechanisms of several iron-scavenging systems are well described, it is not clear whether such systems are targets for selection during adaptation of P. aeruginosa to the host environment. Here we investigated...... the within-host evolution of the transmissible P. aeruginosa DK2 lineage. We found positive selection for promoter mutations leading to increased expression of the phu system. By mimicking conditions of the CF airways in vitro, we experimentally demonstrate that increased expression of phuR confers a growth...

  5. Impact of new water systems on healthcare-associated colonization or infection with Pseudomonas aeruginosa

    Science.gov (United States)

    Lefebvre, Annick; Quantin, Catherine; Vanhems, Philippe; Lucet, Jean-Christophe; Bertrand, Xavier; Astruc, Karine; Chavanet, Pascal; Aho-Glélé, Ludwig S.

    2016-01-01

    Aim: We aimed to study the impact of new water systems, which were less contaminated with P. aeruginosa, on the incidence of healthcare-associated P. aeruginosa cases (colonizations or infections) in care units that moved to a different building between 2005 and 2014. Methods: Generalized Estimated Equations were used to compare the incidence of P. aeruginosa healthcare-associated cases according to the building. Results: Twenty-nine units moved during the study period and 2,759 cases occurred in these units. No difference was observed when the new building was compared with older buildings overall. Conclusion: Our results did not support our hypothesis of a positive association between water system contamination and the incidence of healthcare-associated P. aeruginosa cases. These results must be confirmed by linking results of water samples and patients’ data. PMID:27274443

  6. Investigating the Antibacterial Effects of Plant Extracts on Pseudomonas aeruginosa and Escherichia coli

    Directory of Open Access Journals (Sweden)

    Jahani

    2016-04-01

    Full Text Available Background Scientists are seeking an appropriate alternative method for curing infections caused by resistant bacteria, since drug resistance is continually increasing. Objectives This research aims to discover the function of some medicine plants on pestiferous Pseudomonas aeruginosa and Escherichia coli in humans. Materials and Methods Bacterial strains were obtained from a standard laboratory. The strains of Pseudomonas aeruginosa ATCC27853 and E.coli ATCC25922 bacteria were used for antimicrobial testing of the extractions. Results Our results showed that Teucrium polium extracts have the minimum density of inhibitory for Escherichia coli, 25 ppm, whereas the maximum of this is for Peganum harmala and Prangos ferulaceae with 100 ppm. The lowest minimum concentration inhibitory value of extracts P. harmala, T. polium, T. pratensis and Rumex was found in 25 ppm against P.aeruginosa. Conclusions The results of our study showed that plant extracts have good antibacterial properties against Pseudomonas aeruginosa and Escherichia coli.

  7. Microbial interactions with the cyanobacterium Microcystis aeruginosa and their dependence on temperature

    DEFF Research Database (Denmark)

    Dziallas, Claudia; Grossart, Hans-Peter

    2012-01-01

    cultures changed in a temperature-dependent manner, its quality greatly varied under the same environmental conditions, but with different associated bacterial communities. Furthermore, temperature affected quantity and quality of cell-bound microcystins, whereby interactions between M. aeruginosa...

  8. Characterization of Imipenem Unsusceptible Pseudomonas Aeruginosa Isolates from Inpatients without Carbapenem Treatment

    Institute of Scientific and Technical Information of China (English)

    Yi-hai Gu; Xiao Zhu; Jing-yun Li; Jun Zhang; Qing-yuan Zhou; Yue Ma; Chang-qin Hu; Shao-hong Jin; and Sheng-hui Cui

    2013-01-01

    Objective To identify the risk factors for imipenem resistance development and transmission of clinical Pseudomonas aeruginosa isolates. Methods Thirty-seven imipenem unsusceptible Pseudomonas aeruginosa isolates collected from patients in absence of carbapenem treatment were characterized by antimicrobial susceptibility test, pulsed field gel electrophoresis (PFGE) and carbapenem resistant mechanism analysis. Results Before the collection of imipenem unsusceptible Pseudomonas aeruginosa isolates, the average time of patients treated with more than one antimicrobial (20.0 ± 9.5 days, n=16) was signiifcantly longer than those treated with only one antimicrobial (12.6 ± 4.4 days, n=21;t-test, Welch, t=-2.9004, P Conclusions Our data demonstrated that exposure to non-carbapenem drug classes, especially lfuoroquinolones andβ-lactams, may be important risk factors for the spread of carbapenem resistant Pseudomonas aeruginosa.

  9. Phenotypic shift in Pseudomonas aeruginosa populations from cystic fibrosis lungs after 2-week antipseudomonal treatment

    DEFF Research Database (Denmark)

    Fernández-Barat, Laia; Ciofu, Oana; Kragh, Kasper N

    2017-01-01

    BACKGROUND: The influence of suppressive therapy on the different P. aeruginosa phenotypes harbored in the lungs of cystic fibrosis (CF) patients remains unclear. Our aim was to investigate the phenotypic changes (mucoidy, hypermutability, antibiotic resistance, transcriptomic profiles and biofil...

  10. Phenotypic and genotypic diversity of Pseudomonas aeruginosa strains isolated from hospitals in siedlce (Poland

    Directory of Open Access Journals (Sweden)

    Katarzyna Wolska

    2012-03-01

    Full Text Available A total of 62 Pseudomonas aeruginosa strains isolated from two hospitals in Siedlce (Poland were studied by repetitive element based PCR (rep-PCR using BOX primer. BOX-PCR results revealed the presence of 7 numerous genotypes and 31 unique patterns among isolates. Generally, the strains of P. aeruginosa were characterized by resistance to many antibiotics tested and by differences in serogroups and types of growth on cetrimide agar medium. However, the P. aeruginosa strains isolated from faeces showed much lower phenotypic and genotypic variations in comparison with strains obtained from other clinical specimens. It was observed that genetic techniques supported by phenotypic tests have enabled to conduct a detailed characterization of P. aeruginosa strains isolated from a particular environment at a particular time.

  11. A Novel Antimicrobial Endolysin, LysPA26, against Pseudomonas aeruginosa

    Science.gov (United States)

    Guo, Mingquan; Feng, Chunyan; Ren, Jie; Zhuang, Xuran; Zhang, Yan; Zhu, Yongzhang; Dong, Ke; He, Ping; Guo, Xiaokui; Qin, Jinhong

    2017-01-01

    The global increase in multidrug resistant (MDR) bacteria has led to phage therapy being refocused upon. A novel endolysin, LysPA26, containing a lysozyme-like domain, was screened against Pseudomonas aeruginosa in this study. It had activity against MDR P. aeruginosa without pretreatment with an outer-membrane permeabilizer. LysPA26 could kill up to 4 log units P. aeruginosa in 30 min. In addition, temperature and pH effect assays revealed that LysPA26 had good stability over a broad range of pH and temperatures. Moreover, LysPA26 could kill other Gram-negative bacteria, such as Klebsiella pneumonia, Acinetobacter baumannii and Escherichia coli, but not Gram-positive bacteria. Furthermore, LysPA26 could eliminate P. aeruginosa in biofilm formation. Our current results show that LysPA26 is a new and promising antimicrobial agent for the combat of Gram-negative pathogens. PMID:28289407

  12. Pseudomonas aeruginosa Diversification during Infection Development in Cystic Fibrosis Lungs—A Review

    Science.gov (United States)

    Sousa, Ana Margarida; Pereira, Maria Olívia

    2014-01-01

    Pseudomonas aeruginosa is the most prevalent pathogen of cystic fibrosis (CF) lung disease. Its long persistence in CF airways is associated with sophisticated mechanisms of adaptation, including biofilm formation, resistance to antibiotics, hypermutability and customized pathogenicity in which virulence factors are expressed according the infection stage. CF adaptation is triggered by high selective pressure of inflamed CF lungs and by antibiotic treatments. Bacteria undergo genetic, phenotypic, and physiological variations that are fastened by the repeating interplay of mutation and selection. During CF infection development, P. aeruginosa gradually shifts from an acute virulent pathogen of early infection to a host-adapted pathogen of chronic infection. This paper reviews the most common changes undergone by P. aeruginosa at each stage of infection development in CF lungs. The comprehensive understanding of the adaptation process of P. aeruginosa may help to design more effective antimicrobial treatments and to identify new targets for future drugs to prevent the progression of infection to chronic stages. PMID:25438018

  13. Draft Genome Sequence of Microcystis aeruginosa CACIAM 03, a Cyanobacterium Isolated from an Amazonian Freshwater Environment

    Science.gov (United States)

    Castro, Wendel Oliveira; Lima, Alex Ranieri Jerônimo; Moraes, Pablo Henrique Gonçalves; Siqueira, Andrei Santos; Aguiar, Délia Cristina Figueira; Baraúna, Anna Rafaella Ferreira; Martins, Luisa Carício; Fuzii, Hellen Thais; de Lima, Clayton Pereira Silva; Vianez-Júnior, João Lídio Silva Gonçalves; Nunes, Márcio Roberto Teixeira; Dall'Agnol, Leonardo Teixeira

    2016-01-01

    Given its toxigenic potential, Microcystis aeruginosa is an important bloom-forming cyanobacterium. Here, we present a draft genome and annotation of the strain CACIAM 03, which was isolated from an Amazonian freshwater environment. PMID:27856592

  14. Epidemiology and Ecology of Opportunistic Premise Plumbing Pathogens: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa

    Science.gov (United States)

    BACKGROUND: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa are opportunistic premise plumbing pathogens (OPPPs) that persist and grow in household plumbing, habitats they share with humans. Infections caused by these OPPPs involve individuals with preexis...

  15. Pseudomonas aeruginosa airway infection recruits and modulates neutrophilic myeloid-derived suppressor cells

    Directory of Open Access Journals (Sweden)

    Hasan Halit Öz

    2016-11-01

    Full Text Available Pseudomonas aeruginosa is an opportunistic pathogen that causes infections mainly in patients with cystic fibrosis (CF lung disease. Despite innate and adaptive immune responses upon infection, P. aeruginosa is capable of efficiently escaping host defenses, but the underlying immune mechanisms remain poorly understood. Myeloid-derived suppressor cells (MDSCs are innate immune cells that are functionally characterized by their potential to suppress T- and natural killer (NK-cell responses. Here we demonstrate, using an airway in vivo infection model, that P. aeruginosa recruits and activates neutrophilic MDSCs, which functionally suppress T-cell responses. We further show that the CF gene defect (cystic fibrosis transmembrane conductance regulator, CFTR modulates the functionality, but not the recruitment or generation of neutrophilic MDSCs. Collectively, we define a mechanism by which P. aeruginosa airway infection undermines host immunity by modulating neutrophilic MDSCs in vivo.

  16. Comparison of Antiseptics’ Efficacy on Pseudomonas Aeruginosa, StaphylococcusEpidermidis and Enterobacter Aeruginosa in Hospital of Imam Khomeini (Urmia

    Directory of Open Access Journals (Sweden)

    Fahim Amini

    2012-04-01

    Full Text Available Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 Background and Objectives: Nosocomial infection is the cause of deaths, morbidity, higher costs and increased length of stay in hospitals. Correct and appropriate use of antiseptic and disinfectants play an important role in reducing infections. In this study the efficacy of antiseptics on bacteria causing hospital infections has been studied.Materials and Methods: This study was conducted in the laboratory of Imam Khomeini Hospital of Uremia. In this study the Antimicrobial activity of Descocid, Korsolex basic, Mikrobac forte and persidin 1% was studied against bacteria causing hospital infections such as Enterobacter aeruginosa 1221 (NCTC 10006, Staphylococcus epidermidis (PTCC: 1435 (Cip81.55 and Pseudomonas aeruginosa Strain PAO1. Sensitivities of bacteria were determined by Minimum inhibitory Concentration (MIC and Minimum bactericidal Concentration (MBC antiseptics. In the second stage, the concentration of antiseptics was prepared according to the manufacturer's suggested protocol and the effect of antimicrobial agents were studied at the certain concentration and contact time.Result: All disinfectants (Descocid, Korsolex basic, Mikrobac forte concentration and contact time, Accordance with the manufacturer's brochure, had inhibitory effect on all bacteria. That this is consistent with the manufacturer's brochure. Persidin one percent in concentration of from 2 and 4 V/V % and exposure time 5 minutes could not inhibit the growth of bacterial. But at concentrations of 10 and 20% respectively 15 and 30 minutes exposure time, all three types of bacteria can be inhibited, which is consistent with the manufacturer's claims.Conclusion: In this study, the efficacy of antiseptics was determined with the Micro-dilution method recommended by the NCCLS. Korsolex basic, weakest antiseptics (the highest MIC for the inhibition of three bacteria was determined

  17. Growth of genetically engineered Pseudomonas aeruginosa and Pseudomonas putida in soil and rhizosphere.

    OpenAIRE

    Yeung, K H; Schell, M A; Hartel, P G

    1989-01-01

    The effect of the addition of a recombinant plasmid containing the pglA gene encoding an alpha-1,4-endopolygalacturonase from Pseudomonas solanacearum on the growth of Pseudomonas aeruginosa and Pseudomonas putida in soil and rhizosphere was determined. Despite a high level of polygalacturonase production by genetically engineered P. putida and P. aeruginosa, the results suggest that polygalacturonase production had little effect on the growth of these strains in soil or rhizosphere.

  18. The Role of Newly Discovered Exotoxin (S Toxin) in Pseudomonas aeruginosa Infections

    Science.gov (United States)

    1981-08-01

    Pseudomonas aeruginosa and Enterobacter aerogenes and bilirubin and SGOT of 280 units. On the third day after his initial procedure he was begun on...under halothane anesthesia. A blood culture drawn for a temperature of 104°F was reported as positive for Pseudomonas aeruginosa on the next day (Sth... growth media, culture methods and specific S antisera have all bee described in detail in previous annual reports., S antigen was quantitated by

  19. Phagocytosis of Pseudomonas aeruginosa by polymorphonuclear leukocytes and monocytes: effect of cystic fibrosis serum.

    OpenAIRE

    Thomassen, M J; Demko, C A; Wood, R.E.; Sherman, J. M.

    1982-01-01

    It has been shown previously that serum from chronically infected patients with cystic fibrosis inhibits the phagocytosis of Pseudomonas aeruginosa by both normal and cystic fibrosis alveolar macrophages. In the present study, the ability of peripheral monocytes and polymorphonuclear leukocytes from normal volunteers and cystic fibrosis patients to phagocytize P. aeruginosa was shown not to be inhibited in the presence of serum from cystic fibrosis patients.

  20. Hepatotoxicity of Microcystis aeruginosa Strains Growing as Blooms in Certain Eutrophic Ponds

    OpenAIRE

    Jha, Prabhat N.; Kumar, Anil; Kumar, Ashok; Singh, Dhananjay P.; Sinha, Rajeshwar P.; Tyagi, Madhu B.

    2006-01-01

    Critical assessment of five eutrophicated ponds of Varanasi city (India) revealed the presence of heavy blooms of cyanobacteria consisting mainly of Microcystis aeruginosa. Crude aqueous extracts of blooms as well as laboratory grown M. aeruginosa isolated from three ponds, namely Lakshmikund, Durgakund and Adityanagar showed toxicity in mouse bioassay test. Crude aqueous extracts from these samples caused death of test mice within 1h of administration (i.p.) with a LD50 of 60 mg/kg body weig...

  1. Comparison of Pseudomonas aeruginosa isolates from mink by serotyping and pulsed-field gel electrophoresis

    DEFF Research Database (Denmark)

    Hammer, Anne Sofie; Pedersen, Karl; Andersen, Thomas Holmen

    2003-01-01

    aeruginosa of specific PFGE types were found to cause clusters of outbreaks on several farms within a few weeks of each other. However, PFGE types of strains causing clusters of farm outbreaks changed from year to year. These results suggest that some outbreaks of haemorrhagic pneumonia are caused...... by pathogenic strains of R aeruginosa spread between farms and animals either mechanically, or through feed or water from a common source, rather than by random nosocomial infections with strains from the farm environment....

  2. A diagnostic PCR assay for the detection of an Australian epidemic strain of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Murphy Anna

    2010-07-01

    Full Text Available Abstract Background Chronic lung infection with the bacterium Pseudomonas aeruginosa is one of the hallmarks of cystic fibrosis (CF and is associated with worsening lung function, increased hospitalisation and reduced life expectancy. A virulent clonal strain of P. aeruginosa (Australian epidemic strain I; AES-I has been found to be widespread in CF patients in eastern Australia. Methods Suppression subtractive hybridization (SSH was employed to identify genetic sequences that are present in the AES-I strain but absent from the sequenced reference strain PAO1. We used PCR to evaluate the distribution of several of the AES-I loci amongst a collection of 188 P. aeruginosa isolates which was comprised of 35 AES-I isolates (as determined by PFGE, 78 non-AES-I CF isolates including other epidemic CF strains as well as 69 P. aeruginosa isolates from other clinical and environmental sources. Results We have identified a unique AES-I genetic locus that is present in all 35 AES-I isolates tested and not present in any of the other 153 P. aeruginosa strains examined. We have used this unique AES-I locus to develop a diagnostic PCR and a real-time PCR assay to detect the presence of P. aeruginosa and AES-I in patient sputum samples. Conclusions We have developed diagnostic PCR assays that are 100% sensitive and 100% specific for the P. aeruginosa strain AES-I. We have also shown that Whatman FTA® Elute cards may be used with PCR-based assays to rapidly detect the presence of P. aeruginosa strains in CF sputum.

  3. Class 1 integron and Imipenem Resistance in Clinical Isolates of Pseudomonas aeruginosa: Prevalence and Antibiotic Susceptibility

    Directory of Open Access Journals (Sweden)

    M Milani

    2010-12-01

    Full Text Available Background and Objectives: Pseudomonas aeruginosa is one of the most important causative agents of nosocomial infections especially in ICU and burn units. P. aeruginosa infections are normally difficult to eradicate due to acquired resistance to many antibiotics. Recent appearance of carbapenem resistant P. aeruginosa isolates is considered a major healthcare problem. The present study was conducted to detect class 1 integron and antibiotic susceptibility profiles of imipenem-sensitive and resistant clinical isolates of P. aeruginosa."nMaterials and Methods: Antibiotic susceptibility profiles and minimum inhibitory concentration against imipenem was studied in 160 clinical isolates of P. aeruginosa by disk agar diffusion method and Etest, respectively. Detection of class 1 integron was performed by the PCR method. Demographic and microbiological data were compared between imipenem susceptible and non-susceptible isolates by the SPSS software."nResults: PCR results showed that 90 (56.3% of P. aeruginosa isolates carried class 1 integron. Antibiotic susceptibility results revealed that 93 (58.1% were susceptible and 67 (41.9% were non-susceptible to imipenem. Comparison of antibiotic susceptibility patterns showed high level of drug resistance among imipenem non-susceptible isolates. We found that MDR phenotype, presence of class 1 integron and hospitalization in ICU and burn units were significantly associated with imipenem non-susceptible isolates."nConclusion: The high frequency of imipenem resistance was seen among our P. aeruginosa isolates. Since carbapenems are considered as the last drugs used for treatment of P. aeruginosa infections, it is crucial to screen imipenem non-susceptible isolates in infection control and optimal therapy.

  4. The effect of manuka honey on the structure of Pseudomonas aeruginosa

    OpenAIRE

    2010-01-01

    Abstract The purpose of this study was to investigate the effects of manuka honey on the structural integrity of Pseudomonas aeruginosa ATCC 27853. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of manuka honey for P. aeruginosa were determined by a microtitre plate method, and the survival of bacteria exposed to a bactericidal concentration of manuka honey was monitored. The effect of manuka honey on the structure of the bacteria was in...

  5. Maintenance of Paraoxonase 2 Activity as a Strategy to Attenuate P. Aeruginosa Virulence

    Science.gov (United States)

    2015-12-01

    SUPPLEMENTARY NOTES 14. ABSTRACT The P. aeruginosa signaling/virulence molecule 3OC12 mediates inactivation of the lactonase paraoxonase 2 (PON2) and induces... mediated inactivation at 3OC12 concentrations expected to be present near P. aeruginosa colonies during infection. In mouse primary cell types PON2 was not...as sensitive to 3OC12- mediated inactivation as in human cells. We also discovered that 3OC12 is rapidly hydrolyzed intracellularly by PON2 to 3OC12

  6. Some bacterial parameters influencing the neutrophil oxidative burst response to Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Jensen, E T; Kharazmi, A; Høiby, N;

    1992-01-01

    or biofilms is an important protective mechanism of the microorganisms. We examined the human PMN oxidative burst response to P. aeruginosa in biofilm and in planktonic form. The PMN chemiluminescence response to P. aeruginosa in biofilms was reduced to 30.5-47.5% (p less than 0.04) and the superoxide...... conclude that biofilm bacteria, although able to stimulate the PMN, result in a reduced, suboptimal response leading to lack of efficient eradication of the bacteria in the chronic infection....

  7. Evolutionary insight from whole-genome sequencing of Pseudomonas aeruginosa from cystic fibrosis patients

    DEFF Research Database (Denmark)

    Marvig, Rasmus Lykke; Madsen Sommer, Lea Mette; Jelsbak, Lars;

    2015-01-01

    is suggested to be due to the large genetic repertoire of P. aeruginosa and its ability to genetically adapt to the host environment. Here, we review the recent work that has applied whole-genome sequencing to understand P. aeruginosa population genomics, within-host microevolution and diversity, mutational...... mechanisms, genetic adaptation and transmission events. Finally, we summarize the advances in relation to medical applications and laboratory evolution experiments....

  8. Glutathione exhibits antibacterial activity and increases tetracycline efficacy against Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Glutathione(GSH) plays important roles in pulmonary diseases,and inhaled GSH therapy has been used to treat cystic fibrosis(CF) patients in clinical trials.The results in this report revealed that GSH altered the sensitivity of Pseudomonas aeruginosa to different antibiotics through pathways unrelated to the oxidative stress as generally perceived.In addition,GSH and its oxidized form inhibited the growth of P.aeruginosa.

  9. Removal of bacterial contaminants and antibiotic resistance genes by conventional wastewater treatment processes in Saudi Arabia: Is the treated wastewater safe to reuse for agricultural irrigation?

    KAUST Repository

    Aljassim, Nada I.

    2015-04-01

    This study aims to assess the removal efficiency of microbial contaminants in a local wastewater treatment plant over the duration of one year, and to assess the microbial risk associated with reusing treated wastewater in agricultural irrigation. The treatment process achieved 3.5 logs removal of heterotrophic bacteria and up to 3.5 logs removal of fecal coliforms. The final chlorinated effluent had 1.8×102 MPN/100mL of fecal coliforms and fulfils the required quality for restricted irrigation. 16S rRNA gene-based high-throughput sequencing showed that several genera associated with opportunistic pathogens (e.g. Acinetobacter, Aeromonas, Arcobacter, Legionella, Mycobacterium, Neisseria, Pseudomonas and Streptococcus) were detected at relative abundance ranging from 0.014 to 21 % of the total microbial community in the influent. Among them, Pseudomonas spp. had the highest approximated cell number in the influent but decreased to less than 30 cells/100mL in both types of effluent. A culture-based approach further revealed that Pseudomonas aeruginosa was mainly found in the influent and non-chlorinated effluent but was replaced by other Pseudomonas spp. in the chlorinated effluent. Aeromonas hydrophila could still be recovered in the chlorinated effluent. Quantitative microbial risk assessment (QMRA) determined that only chlorinated effluent should be permitted for use in agricultural irrigation as it achieved an acceptable annual microbial risk lower than 10-4 arising from both P. aeruginosa and A. hydrophila. However, the proportion of bacterial isolates resistant to 6 types of antibiotics increased from 3.8% in the influent to 6.9% in the chlorinated effluent. Examples of these antibiotic-resistant isolates in the chlorinated effluent include Enterococcus and Enterobacter spp. Besides the presence of antibiotic-resistant bacterial isolates, tetracycline resistance genes tetO, tetQ, tetW, tetH, tetZ were also present at an average 2.5×102, 1.6×102, 4.4×102, 1

  10. Starch removal from potato tuber sections.

    Science.gov (United States)

    Fronda, A; Jona, R

    1991-01-01

    Heating plant sections at 90 C with 0.5% aqueous ammonium oxalate is required to remove pectins. When applied to tissues rich in starch such as potato, this step produces heavy dextrinization of the starch which hinders subsequent evaluation of the extinction values of the cell walls. To overcome this a method has been devised to brush away the starch granules from the sections with a thin paint brush, just after paraffin removal by xylene. The slide is then processed as usual: pectins are removed by heat treatment, cell walls are stained with PAS and the stain intensity can be evaluated by photometry.

  11. Carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii in the nosocomial setting in Latin America.

    Science.gov (United States)

    Labarca, Jaime A; Salles, Mauro José Costa; Seas, Carlos; Guzmán-Blanco, Manuel

    2016-01-01

    Increasing prevalence of carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter baumannii strains in the nosocomial setting in Latin America represents an emerging challenge to public health, as the range of therapeutic agents active against these pathogens becomes increasingly constrained. We review published reports from 2002 to 2013, compiling data from throughout the region on prevalence, mechanisms of resistance and molecular epidemiology of carbapenem-resistant strains of P. aeruginosa and A. baumannii. We find rates of carbapenem resistance up to 66% for P. aeruginosa and as high as 90% for A. baumannii isolates across the different countries of Latin America, with the resistance rate of A. baumannii isolates greater than 50% in many countries. An outbreak of the SPM-1 carbapenemase is a chief cause of resistance in P. aeruginosa strains in Brazil. Elsewhere in Latin America, members of the VIM family are the most important carbapenemases among P. aeruginosa strains. Carbapenem resistance in A. baumannii in Latin America is predominantly due to the oxacillinases OXA-23, OXA-58 and (in Brazil) OXA-143. Susceptibility of P. aeruginosa and A. baumannii to colistin remains high, however, development of resistance has already been detected in some countries. Better epidemiological data are needed to design effective infection control interventions.

  12. A universal primer multiplex PCR method for typing of toxinogenic Pseudomonas aeruginosa.

    Science.gov (United States)

    Shi, Hui; Trinh, Quoclinh; Xu, Wentao; Zhai, Baiqiang; Luo, Yunbo; Huang, Kunlun

    2012-09-01

    Pseudomonas aeruginosa is a well-known opportunistic pathogen that can cause acute nosocomial necrotizing pneumonia and genetic disorder cystic fibrosis of lung patients. Pathogenic interactions between P. aeruginosa and hosts are often guided by the secreted virulence determinants that interact with specific host targets. Exotoxin A, pyocyanin, elastase, and type III secretion system are the most significant virulence determinants and cause great concern. However, P. aeruginosa in various environments has high genotypic diversity, leading to deficiency of exotoxin genes for some P. aeruginosa strains. In current study, a universal primer-multiplex PCR method (UP-MPCR) was employed for the detection of five significant enterotoxin genes (toxA, phzM, lasB, ExoU, and ExoS) and one internal control gene ecfX in P. aeruginosa. Owing to the application of universal primer (UP), different targeted products have identical amplified efficiency and the sensitivity of multiplex PCR is improved. In addition, the complexity of multiplex PCR system is reduced and the compatibility of primers in a reaction is greatly increased. This UP-MPCR method can detect the presence of five P. aeruginosa enterotoxin genes in a single assay more rapidly and sensitively than conventional methods. In 214 drinking water and environmental isolates, the ExoU, ExoS, phzM, toxA, and lasB genes were detected in 20 (9 %), 180 (84 %), 179 (84 %), 196 (92 %), and 171 (80 %) isolates, respectively.

  13. Pseudomonas aeruginosa in Swimming Pool Water: Evidences and Perspectives for a New Control Strategy

    Directory of Open Access Journals (Sweden)

    Marco Guida

    2016-09-01

    Full Text Available Pseudomonas aeruginosa is frequently isolated in swimming pool settings. Nine recreational and rehabilitative swimming pools were monitored according to the local legislation. The presence of P. aeruginosa was correlated to chlorine concentration. The ability of the isolates to form a biofilm on plastic materials was also investigated. In 59.5% of the samples, microbial contamination exceeded the threshold values. P. aeruginosa was isolated in 50.8% of these samples. The presence of P. aeruginosa was not correlated with free or total chlorine amount (R2 < 0.1. All the isolates were moderate- to strong-forming biofilm (Optical Density O.D.570 range 0.7–1.2. To control biofilm formation and P. aeruginosa colonization, Quantum FreeBioEnergy© (QFBE, FreeBioEnergy, Brisighella, Italy, has been applied with encouraging preliminary results. It is a new, promising control strategy based on the change of an electromagnetic field which is responsible for the proliferation of some microorganisms involved in biofilm formation, such as P. aeruginosa.

  14. Evolution and adaptation in Pseudomonas aeruginosa biofilms driven by mismatch repair system-deficient mutators.

    Science.gov (United States)

    Luján, Adela M; Maciá, María D; Yang, Liang; Molin, Søren; Oliver, Antonio; Smania, Andrea M

    2011-01-01

    Pseudomonas aeruginosa is an important opportunistic pathogen causing chronic airway infections, especially in cystic fibrosis (CF) patients. The majority of the CF patients acquire P. aeruginosa during early childhood, and most of them develop chronic infections resulting in severe lung disease, which are rarely eradicated despite intensive antibiotic therapy. Current knowledge indicates that three major adaptive strategies, biofilm development, phenotypic diversification, and mutator phenotypes [driven by a defective mismatch repair system (MRS)], play important roles in P. aeruginosa chronic infections, but the relationship between these strategies is still poorly understood. We have used the flow-cell biofilm model system to investigate the impact of the mutS associated mutator phenotype on development, dynamics, diversification and adaptation of P. aeruginosa biofilms. Through competition experiments we demonstrate for the first time that P. aeruginosa MRS-deficient mutators had enhanced adaptability over wild-type strains when grown in structured biofilms but not as planktonic cells. This advantage was associated with enhanced micro-colony development and increased rates of phenotypic diversification, evidenced by biofilm architecture features and by a wider range and proportion of morphotypic colony variants, respectively. Additionally, morphotypic variants generated in mutator biofilms showed increased competitiveness, providing further evidence for mutator-driven adaptive evolution in the biofilm mode of growth. This work helps to understand the basis for the specific high proportion and role of mutators in chronic infections, where P. aeruginosa develops in biofilm communities.

  15. Pseudomonas aeruginosa and Heterotrophic Bacteria Count in Bottled Waters in Iran

    Directory of Open Access Journals (Sweden)

    Matin MOHAMMADI KOUCHESFAHANI

    2015-11-01

    Full Text Available Background: Nowadays, due to increased public awareness about water pollution and water borne diseases as well as water network deficiencies, bottled water consumers have increased dramatically worldwide, including Iran. Pseudomonas aeruginosa is an opportunistic human pathogen capable of causing widespread infections in burn and immune-compromised patients. The aim of this study was to investigate, P. aeruginosa in bottled waters selling in Iranian markets.Methods: One hundred and twenty samples of five unknown (not famous domestic bottled water brands were purchased from Tehran retailers during 2013. The samples were evaluated for the presence of P. aeruginosa. In addition, heterotrophic plate counts were determined by incubation at 37 °C for 24 h.Results: P. aeruginosa was detected in 36.7% (44 samples of all samples examined. In addition, heterotrophic bacteria in 32.5% (39 samples of the samples were higher than 100 CFU/mL, while in 7.5% (9 samples of the samples HPC relied between 20 and 100 CFU/ml.Conclusion: In contrast to public believe, bottled waters are not free of microorganisms, and it is suggested that authorities should provide stricter monitoring and control plan for water resources and plants. Concerning HPC and P. aeruginosa brands B and D were not suitable for drinking. Keywords: Heterotrophic plate count, Pseudomonas aeruginosa, Bottled water

  16. Phenolic compounds affect production of pyocyanin, swarming motility and biofilm formation of Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    Aylin Ugurlu; Aysegul Karahasan Yagci; Seyhan Ulusoy; Burak Aksu; Gulgun Bosgelmez-Tinaz

    2016-01-01

    Objective: To investigate the effects of plant-derived phenolic compounds (i.e. caffeic acid, cinnamic acid, ferulic acid and vanillic acid) on the production of quorum sensing regulated virulence factors such as pyocyanin, biofilm formation and swarming motility of Pseudomonas aeruginosa (P. aeruginosa) isolates. Methods: Fourteen clinical P. aeruginosa isolates obtained from urine samples and P. aeruginosa PA01 strain were included in the study. The antibacterial effects of phenolic compounds were screened by well diffusion assay. Pyocyanin and biofilm ac-tivity were measured from culture supernatants and the absorbance values were measured using a spectrophotometer. Swarming plates supplemented with phenolic acids were point inoculated with P. aeruginosa strains and the ability to swarm was determined by measuring the distance of swarming from the central inoculation site. Results: Tested phenolic compounds reduced the production of pyocyanin and biofilm formation without affecting growth compared to untreated cultures. Moreover, these compounds blocked about 50% of biofilm production and swarming motility in P. aeruginosa isolates. Conclusions: We may suggest that if swarming and consecutive biofilm formation could be inhibited by the natural products as shown in our study, the bacteria could not attach to the surfaces and produce chronic infections. Antimicrobials and natural products could be combined and the dosage of antimicrobials could be reduced to overcome antimicrobial resistance and drug side effects.

  17. Phenolic compounds affect production of pyocyanin, swarming motility and biofilm formation of Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    Aylin Ugurlu; Aysegul Karahasan Yagci; Seyhan Ulusoy; Burak Aksu; Gulgun Bosgelmez-Tinaz

    2016-01-01

    Objective: To investigate the effects of plant-derived phenolic compounds(i.e. caffeic acid, cinnamic acid, ferulic acid and vanillic acid) on the production of quorum sensing regulated virulence factors such as pyocyanin, biofilm formation and swarming motility of Pseudomonas aeruginosa(P. aeruginosa) isolates.Methods: Fourteen clinical P. aeruginosa isolates obtained from urine samples and P. aeruginosa PA01 strain were included in the study. The antibacterial effects of phenolic compounds were screened by well diffusion assay. Pyocyanin and biofilm activity were measured from culture supernatants and the absorbance values were measured using a spectrophotometer. Swarming plates supplemented with phenolic acids were point inoculated with P. aeruginosa strains and the ability to swarm was determined by measuring the distance of swarming from the central inoculation site.Results: Tested phenolic compounds reduced the production of pyocyanin and biofilm formation without affecting growth compared to untreated cultures. Moreover, these compounds blocked about 50% of biofilm production and swarming motility in P. aeruginosa isolates.Conclusions: We may suggest that if swarming and consecutive biofilm formation could be inhibited by the natural products as shown in our study, the bacteria could not attach to the surfaces and produce chronic infections. Antimicrobials and natural products could be combined and the dosage of antimicrobials could be reduced to overcome antimicrobial resistance and drug side effects.

  18. Photodynamic antimicrobial therapy to inhibit pseudomonas aeruginosa of corneal isolates (Conference Presentation)

    Science.gov (United States)

    Durkee, Heather A.; Relhan, Nidhi; Arboleda, Alejandro; Halili, Francisco; De Freitas, Carolina; Alawa, Karam; Aguilar, Mariela C.; Amescua, Guillermo; Miller, Darlene; Parel, Jean-Marie

    2016-03-01

    Keratitis associated with Pseudomonas aeruginosa is difficult to manage. Treatment includes antibiotic eye drops, however, some strains of Pseudomonas aeruginosa are resistant. Current research efforts are focused on finding alternative and adjunct therapies to treat multi-drug resistant bacteria. One promising alternate technique is photodynamic therapy (PDT). The purpose of this study was to evaluate the effect of riboflavin- and rose bengal-mediated PDT on Pseudomonas aeruginosa keratitis isolates in vitro. Two isolates (S+U- and S-U+) of Pseudomonas aeruginosa were derived from keratitis patients and exposed to five experimental groups: (1) Control (dark, UV-A irradiation, 525nm irradiation); (2) 0.1% riboflavin (dark, UV-A irradiation); and (3) 0.1% rose bengal, (4) 0.05% rose bengal and (5) 0.01% rose bengal (dark, 525nm irradiation). Three days after treatment, in dark conditions of all concentration of riboflavin and rose bengal showed no inhibition in both S+U- and S-U+ strains of Pseudomonas aeruginosa. In 0.1% and 0.05% rose bengal irradiated groups, for both S+U- and S-U+ strains, there was complete inhibition of bacterial growth in the central 50mm zone corresponding to the diameter of the green light source. These in vitro results suggest that rose bengal photodynamic therapy may be an effective adjunct treatment for Pseudomonas aeruginosa keratitis.

  19. ANTIBACTERIAL ACTIVITY OF TRADITIONAL HERBS AND STANDARD ANTIBIOTICS AGAINST POULTRY ASSOCIATED PSEUDOMONAS AERUGINOSA

    Directory of Open Access Journals (Sweden)

    Affia Rafique

    2012-10-01

    Full Text Available Present study aims to access the antibacterial activity of medicinal plants and antibiotics against poultry associated Pseudomonas aeruginosa. P. aeruginosa is the most widespread avian pathogen and it produces a range of toxins and enzymes that may contribute to pathogenicity. P. aeruginosa was isolated from the chicken liver and identified through biochemical methods. The antibacterial activity of extracts of medicinal herbs and various antibiotics were analyzed against P. aeruginosa through agar disc diffusion method. P. aeruginosa was susceptible against Norfloxacin, Chloramphenicol, Streptomycin, Gentamicin, Tobramycin, and Ciprofloxacin. Whereas, moderately susceptible in case of Oxytetracycline, Neomycin, Lincomycin, and Sulfomethoxyzol. It was also analyzed that Ampicillin, Tetracycline, Penicillin G and Trimethoprim had no effect. Among the plants tested C. zylanicum, C. cyminum, T. ammi, S. aromaticum and green part of M. charantia were most active. The maximum antibacterial activity was calculated by the extracts of isoamylalcohol of C. zylanicum, C. cyminum, T. ammi, S. aromaticum, and ethanolic and methanol extract of green part of M. charantia against P. aeruginosa. This study indicated that these medicinal plants could be the potential source for antimicrobial agents. Hence, these medicinal plants can be further subjected to isolation of the therapeutic antimicrobials and further pharmacological evaluation.

  20. Evolution and adaptation in Pseudomonas aeruginosa biofilms driven by mismatch repair system-deficient mutators.

    Directory of Open Access Journals (Sweden)

    Adela M Luján

    Full Text Available Pseudomonas aeruginosa is an important opportunistic pathogen causing chronic airway infections, especially in cystic fibrosis (CF patients. The majority of the CF patients acquire P. aeruginosa during early childhood, and most of them develop chronic infections resulting in severe lung disease, which are rarely eradicated despite intensive antibiotic therapy. Current knowledge indicates that three major adaptive strategies, biofilm development, phenotypic diversification, and mutator phenotypes [driven by a defective mismatch repair system (MRS], play important roles in P. aeruginosa chronic infections, but the relationship between these strategies is still poorly understood. We have used the flow-cell biofilm model system to investigate the impact of the mutS associated mutator phenotype on development, dynamics, diversification and adaptation of P. aeruginosa biofilms. Through competition experiments we demonstrate for the first time that P. aeruginosa MRS-deficient mutators had enhanced adaptability over wild-type strains when grown in structured biofilms but not as planktonic cells. This advantage was associated with enhanced micro-colony development and increased rates of phenotypic diversification, evidenced by biofilm architecture features and by a wider range and proportion of morphotypic colony variants, respectively. Additionally, morphotypic variants generated in mutator biofilms showed increased competitiveness, providing further evidence for mutator-driven adaptive evolution in the biofilm mode of growth. This work helps to understand the basis for the specific high proportion and role of mutators in chronic infections, where P. aeruginosa develops in biofilm communities.

  1. Pooled human immunoglobulins reduce adhesion of Pseudomonas aeruginosa in a parallel plate flow chamber.

    Science.gov (United States)

    Poelstra, K A; van der Mei, H C; Gottenbos, B; Grainger, D W; van Horn, J R; Busscher, H J

    2000-08-01

    The influence of pooled polyclonal immunoglobulin (IgG) interactions with both bacteria and model substrates in altering Pseudomonas aeruginosa surface adhesion is reported. Opsonization of this pathogen by polyclonal human IgG and preadsorption of IgG to glass surfaces both effectively reduce initial deposition rates and surface growth of P. aeruginosa IFO3455 from dilute nutrient broth in a parallel plate flow chamber. Polyclonal IgG depleted of P. aeruginosa-specific antibodies reduces the initial deposition rate or surface growth to levels intermediate between exposed and nonexposed IgG conditions. Bacterial surface properties are changed in the presence of opsonizing IgG. Plateau contact angle analysis via sessile drop technique shows a drop in P. aeruginosa surface hydrophobicity after IgG exposure consistent with a more hydrophilic IgG surface coat. Zeta potential values for opsonized versus nonopsonized bacteria exhibit little change. X-ray photoelectron spectroscopy measurements provide surface compositional evidence for IgG attachment to bacterial surfaces. Surface elemental ratios attributed to IgG protein signals versus those attributed primarily to bacterial polysaccharide surface or lipid membrane change with IgG opsonization. Direct evidence for antibody-modified P. aeruginosa surface properties correlates both with reduction of bacterial adhesion to glass surfaces under flow in nutrient medium reported and previous reports of IgG efficacy against P. aeruginosa motility in vitro and infection in vivo.

  2. Immunological evaluation of an alginate-based conjugate as a vaccine candidate against Pseudomonas aeruginosa.

    Science.gov (United States)

    Farjah, Ali; Owlia, Parviz; Siadat, Seyed Davar; Mousavi, Seyed Fazlollah; Ardestani, Mehdi Shafiee; Mohammadpour, Hashem Khorsand

    2015-02-01

    Pseudomonas aeruginosa is an opportunistic pathogen that causes serious infections, is usually resistant to antimicrobial agents, and is the leading cause of morbidity and premature mortality in patients with cystic fibrosis (CF). Mucoid strains of P. aeruginosa produce a virulence factor known as alginate. Developing a strategy to raise opsonic antibodies against alginate could be promising for the treatment of P. aeruginosa infection in CF patients. Conjugation of alginate to a carrier protein is a good method for increasing the immunogenicity of alginate. We conjugated alginate to the outer membrane vesicle (OMV) of Neisseria meningitidis serogroup B, which is a safe carrier protein, and evaluated its efficacy in mice. To evaluate the immune response, total IgG, IgG1, IgG2a, and IgG2b titers were analyzed. Immunization of mice with the alginate-OMV conjugate raised the levels of opsonic antibodies, and the vaccinated mice were protected when challenged intranasally with P. aeruginosa. Further studies showed that the conjugated vaccine could eliminate P. aeruginosa from the lungs of infected mice. This study supports the proposal that immunization of mice with an alginate-OMV conjugate vaccine could be safe and protective against P. aeruginosa infection.

  3. Effect of Tyrosol and Farnesol on Virulence and Antibiotic Resistance of Clinical Isolates of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Shaymaa Hassan Abdel-Rhman

    2015-01-01

    Full Text Available Mixed-species biofilms could create a protected environment that allows for survival to external antimicrobials and allows different bacterial-fungal interactions. Pseudomonas aeruginosa-Candida albicans coexistence is an example for such mixed-species community. Numerous reports demonstrated how P. aeruginosa or its metabolites could influence the growth, morphogenesis, and virulence of C. albicans. In this study, we investigated how the C. albicans quorum sensing compounds, tyrosol and farnesol, might affect Egyptian clinical isolates of P. aeruginosa regarding growth, antibiotic sensitivity, and virulence. We could demonstrate that tyrosol possesses an antibacterial activity against P. aeruginosa (10 µM inhibited more than 50% of growth after 16 h cultivation. Moreover, we could show for the first time that tyrosol strongly inhibits the production of the virulence factors hemolysin and protease in P. aeruginosa, whereas farnesol inhibits, to lower extent, hemolysin production in this bacterial pathogen. Cumulatively, tyrosol is expected to strongly affect P. aeruginosa in mixed microbial biofilm.

  4. Structure of the Pseudomonas aeruginosa Type IVa Pilus Secretin at 7.4 Å.

    Science.gov (United States)

    Koo, Jason; Lamers, Ryan P; Rubinstein, John L; Burrows, Lori L; Howell, P Lynne

    2016-10-04

    Type IVa pili (T4aP) function as bacterial virulence factors. T4aP pass through the outer membranes of Gram-negative bacteria via homo-oligomeric secretins. We present a 7.4 Å cryoelectron microscopy structure of the Pseudomonas aeruginosa PilQ secretin. Peripheral and internal features show that the secretin is composed of 14 subunits with C7 symmetry. The channel is a ribbed cylinder with central peripheral spokes and a central gate closed on the periplasmic side. The structure suggests that during pilus extrusion, the central gate is displaced to the interior walls and that no additional conformational changes are required, as the internal diameter can accommodate the pilus. The N1 domain was resolved, while the N0 and the N-terminal β-domains proposed to bind peptidoglycan were absent in class average images and the final 3D map, indicating a high flexibility. These data provide the highest-resolution structure to date of a T4aP secretin.

  5. Antibiotic resistance in Pseudomonas aeruginosa biofilms: towards the development of novel anti-biofilm therapies.

    Science.gov (United States)

    Taylor, Patrick K; Yeung, Amy T Y; Hancock, Robert E W

    2014-12-10

    The growth of bacteria as structured aggregates termed biofilms leads to their protection from harsh environmental conditions such as physical and chemical stresses, shearing forces, and limited nutrient availability. Because of this highly adapted ability to survive adverse environmental conditions, bacterial biofilms are recalcitrant to antibiotic therapies and immune clearance. This is particularly problematic in hospital settings where biofilms are a frequent cause of chronic and device-related infections and constitute a significant burden on the health-care system. The major therapeutic strategy against infections is the use of antibiotics, which, due to adaptive resistance, are often insufficient to clear biofilm infections. Thus, novel biofilm-specific therapies are required. Specific features of biofilm development, such as surface adherence, extracellular matrix formation, quorum sensing, and highly regulated biofilm maturation and dispersal are currently being studied as targets to be exploited in the development of novel biofilm-specific treatments. Using Pseudomonas aeruginosa for illustrative purposes, this review highlights the antibiotic resistance mechanisms of biofilms, and discusses current research into novel biofilm-specific therapies.

  6. Clusters of genetically similar isolates of Pseudomonas aeruginosa from multiple hospitals in the UK.

    Science.gov (United States)

    Martin, Kate; Baddal, Buket; Mustafa, Nazim; Perry, Claire; Underwood, Anthony; Constantidou, Chrystala; Loman, Nick; Kenna, Dervla T; Turton, Jane F

    2013-07-01

    Variable number tandem repeat (VNTR) analysis at nine loci of isolates of Pseudomonas aeruginosa submitted to the national reference laboratory from UK hospitals, from over 2000 patients, between June 2010 and June 2012 revealed four widely found types that collectively were received from approximately a fifth of patients, including from those with cystic fibrosis. These types were also prevalent among related submissions from the clinical environment and were received from up to 54 (out of 143) hospitals. Multi-locus sequence typing and blaOXA-50-like sequencing confirmed the clonal relationship within each cluster, and representatives from multiple centres clustered within about 70 % by pulsed-field gel electrophoresis. Illumina sequencing of 12 isolates of cluster A of VNTR profile 8, 3, 4, 5, 2, 3, 5, 2, x (where the repeat number at the last, most discriminatory locus is variable) revealed a large number of variably present targets in the accessory genome and seven of these were sought by PCR among a larger set of isolates. Representatives from patients within a single centre mostly had distinct accessory gene profiles, suggesting that these patients acquired the strain independently, while those with clear epidemiological links shared the same profile. Profiles also varied between representatives from different centres. Epidemiological investigations of widely found types such as these require the use of finer-typing methods, which increasingly will be informed by next generation sequencing.

  7. Effect of Silver Nanoparticles Against the Formation of Biofilm by Pseudomonas aeruginosa an In silico Approach.

    Science.gov (United States)

    Vyshnava, Satyanarayana Swamy; Kanderi, Dileep Kumar; Panjala, Shiva Prasad; Pandian, Kamesh; Bontha, Rajasekhar Reddy; Goukanapalle, Praveen Kumar Reddy; Banaganapalli, Babajan

    2016-10-01

    Studies were undertaken to examine the mechanism of mediation of silver nanoparticles in inhibiting biofilm formation by Pseudomonas aeruginosa through LuxI/LuxR system of signal transduction. This study includes the basic signaling transduction mechanism LasR, QscR, RhlR, and Vfr signaling model systems. The arbitrary homology models built with the I-TASSER server were evaluated and validated with the Qmean web server. Based on the Z-score and the relative square mean distance (RMSD) values, the structures were validated. The interaction results of the nanoparticle with the rigid docking proved the requirement of minimal energy for the inhibition of the protein active site by the silver nanoparticle. This principle docking experiment suggests that the biofilm formation in Gram-negative bacteria can be inhibited by the silver nanoparticles at the signal transduction level. Graphical abstract Systematic outline of present study; Stage one provides the data sampling and generation of pdb systems to conform the structure of bacterial signal sytems like LasR/LasI; RhlR/RhrI; QscR/QscI; VfrR/VfrI. Stage two involves docking of silver nanoparticles with Bacterial signal protein strucutres which are listed in Stage one. The Final Stage involves in understanding the development of appropriate mechanism behind the biofilm inhibition by silver nanoparticles.

  8. Rational Design of Potent and Selective Inhibitors of an Epoxide Hydrolase Virulence Factor from Pseudomonas aeruginosa.

    Science.gov (United States)

    Kitamura, Seiya; Hvorecny, Kelli L; Niu, Jun; Hammock, Bruce D; Madden, Dean R; Morisseau, Christophe

    2016-05-26

    The virulence factor cystic fibrosis transmembrane conductance regulator (CFTR) inhibitory factor (Cif) is secreted by Pseudomonas aeruginosa and is the founding member of a distinct class of epoxide hydrolases (EHs) that triggers the catalysis-dependent degradation of the CFTR. We describe here the development of a series of potent and selective Cif inhibitors by structure-based drug design. Initial screening revealed 1a (KB2115), a thyroid hormone analog, as a lead compound with low micromolar potency. Structural requirements for potency were systematically probed, and interactions between Cif and 1a were characterized by X-ray crystallography. On the basis of these data, new compounds were designed to yield additional hydrogen bonding with residues of the Cif active site. From this effort, three compounds were identified that are 10-fold more potent toward Cif than our first-generation inhibitors and have no detectable thyroid hormone-like activity. These inhibitors will be useful tools to study the pathological role of Cif and have the potential for clinical application.

  9. Staphylococcus aureus Alters Growth Activity, Autolysis, and Antibiotic Tolerance in a Human Host-Adapted Pseudomonas aeruginosa Lineage

    DEFF Research Database (Denmark)

    Frydenlund Michelsen, Charlotte; Christensen, Anne-Mette; Bojer, Martin Saxtorph

    2014-01-01

    Interactions among members of polymicrobial infections or between pathogens and the commensal flora may determine disease outcomes. Pseudomonas aeruginosa and Staphylococcus aureus are important opportunistic human pathogens and are both part of the polymicrobial infection communities in human....... aeruginosa DK2 strains outcompeted S. aureus during coculture on agar plates, we found that later P. aeruginosa DK2 strains showed a commensal-like interaction, where S. aureus was not inhibited by P. aeruginosa and the growth activity of P. aeruginosa was enhanced in the presence of S. aureus. This effect...... is mediated by one or more extracellular S. aureus proteins greater than 10 kDa, which also suppressed P. aeruginosa autolysis and prevented killing by clinically relevant antibiotics through promoting small-colony variant (SCV) formation. The commensal interaction was abolished with S. aureus strains mutated...

  10. Surto de abscesso mandibular por Pseudomonas aeruginosa em ovinos

    Directory of Open Access Journals (Sweden)

    Rogério Martins Amorim

    2011-09-01

    Full Text Available Descreve-se um surto de abscesso mandibular em ovelhas da raça Bergamácia no município de Botucatu, estado de São Paulo. Do rebanho de 120 animais, 35 apresentaram aumento de volume mandibular com a presença de nódulos únicos, de consistência pétrea, de diferentes tamanhos, fistulados ou não e sem indicativos de inflamação dos tecidos moles adjacentes. Os animais eram criados em pasto de Panicum maximum cv. Tanzânia com água e sal mineral ad libitum e everminados, via oral, com pistolas dosificadoras. O material para diagnóstico microbiológico e antibiograma foi coletado de cinco animais acometidos, por punção e aspiração dos nódulos. Dos 35 animais acometidos, 19 foram submetidos ao exame radiográfico, um ao exame tomográfico e outro à biópsia óssea da região submandibular. O único ovino que morreu, encontrava-se em estado de caquexia provavelmente devido à localização do aumento de volume que afetou a implantação dos dentes molares daquela região impedindo a apreensão e mastigação adequadas levando a perda da condição corporal e morte. Ao exame necroscópico, observaram-se áreas de necrose caseosa na mandíbula direita de onde isolou-se Pseudomonas aeruginosa. O tratamento utilizado foi baseado na aplicação de iodeto de sódio a 10% por via intramuscular e antibioticoterapia segundo antibiograma com enrofloxacina por via intramuscular, porém com pouca eficácia. Diante do quadro clínico, dos dados de anamnese, da localização das lesões no tecido ósseo mandibular, do resultado do cultivo microbiológico, das alterações radiográficas e tomográficas foi feito o diagnóstico de abscesso mandibular causado por Pseudomonas aeruginosa.

  11. Pseudomonas aeruginosa Quorum-Sensing Systems May Control Virulence Factor Expression in the Lungs of Patients with Cystic Fibrosis

    OpenAIRE

    Erickson, David L.; Endersby, Ryan; Kirkham, Amanda; Stuber, Kent; Vollman, Dolina D.; Rabin, Harvey R; Mitchell, Ian; Storey, Douglas G.

    2002-01-01

    Individuals with cystic fibrosis (CF) are commonly colonized with Pseudomonas aeruginosa. The chronic infections caused by P. aeruginosa are punctuated by acute exacerbations of the lung disease, which lead to significant morbidity and mortality. As regulators of virulence determinants, P. aeruginosa quorum-sensing systems may be active in the chronic lung infections associated with CF. We have examined the levels of autoinducer molecules and transcript accumulation from the bacterial populat...

  12. Outbreak of gut colonization by Pseudomonas aeruginosa in immunocompromised children undergoing total digestive decontamination: analysis by pulsed-field electrophoresis.

    OpenAIRE

    J. Boukadida; De Montalembert, M.; Gaillard, J L; Gobin, J; Grimont, F.; Girault, D; Véron, M; Berche, P

    1991-01-01

    We analyzed an outbreak of gut colonization by Pseudomonas aeruginosa occurring in an intensive care hematology unit by using conventional typing methods and pulsed-field electrophoresis. In October and November 1989, the feces of four immunocompromised children undergoing total digestive decontamination were colonized by P. aeruginosa. Ten isolates were obtained from the gut flora in pure culture. Retrospective investigations found that one P. aeruginosa isolate from stools of one of the pat...

  13. Detection of extended spectrum beta lactamases, ampc beta lactamases and metallobetalactamases in clinical isolates of ceftazidime resistant Pseudomonas Aeruginosa

    Directory of Open Access Journals (Sweden)

    Sivaraman Umadevi

    2011-12-01

    Full Text Available We studied the prevalence of ceftazidime resistance in Pseudomonas aeruginosa and the rates of extended-spectrum β-lactamase (ESBL, AmpC β-lactamase (AmpC and metallo-β-lactamase (MBL production among the ceftazidime resistant Pseudomonas aeruginosa. A very high rate of MBL production was observed, which suggested it to be an important contributing factor for ceftazidime resistance among Pseudomonas aeruginosa.

  14. Synergistic Activities of an Efflux Pump Inhibitor and Iron Chelators against Pseudomonas aeruginosa Growth and Biofilm Formation

    DEFF Research Database (Denmark)

    Liu, Yang; Yang, Liang; Molin, Søren

    2010-01-01

    The efflux pump inhibitor phenyl-arginine-beta-naphthylamide (PA beta N) was paired with iron chelators 2,2'-dipyridyl, acetohydroxamic acid, and EDTA to assess synergistic activities against Pseudomonas aeruginosa growth and biofilm formation. All of the tested iron chelators synergistically...... inhibited P. aeruginosa growth and biofilm formation with PA beta N. PA beta N-EDTA showed the most promising activity against P. aeruginosa growth and biofilm formation....

  15. Reactor for removing ammonia

    Science.gov (United States)

    Luo, Weifang; Stewart, Kenneth D.

    2009-11-17

    Disclosed is a device for removing trace amounts of ammonia from a stream of gas, particularly hydrogen gas, prepared by a reformation apparatus. The apparatus is used to prevent PEM "poisoning" in a fuel cell receiving the incoming hydrogen stream.

  16. Nephrectomy (Kidney Removal)

    Science.gov (United States)

    ... if your entire kidney needs to be removed. Robot-assisted laparoscopic surgery. In a variation of laparoscopic ... living kidney donor. Rochester, Minn.: Mayo Foundation for Medical Education and Research; 2014. AskMayoExpert. Partial nephrectomy. Rochester, ...

  17. Asbestos Removal Case History.

    Science.gov (United States)

    Haney, Stanley J.

    1986-01-01

    The engineer for a California school district describes the asbestos removal from the ceilings of El Camino High School. Discusses forming a design team, use of consultants, specifications, relations with contractors, and staff notification. (MLF)

  18. Breast lump removal

    Science.gov (United States)

    ... cannot feel it when examining you, a wire localization will be done before the surgery. A radiologist ... send the lump to a laboratory for more testing. Why the Procedure is Performed Surgery to remove ...

  19. Research of possibilities for the use of multienzyme complex of algae Mycrocystis aeruginosa for treatment of drinkable and process water

    Directory of Open Access Journals (Sweden)

    V. I. Karpenko

    2010-01-01

    Full Text Available The methods and a laboratory bench have been developed for research of the multienzyme systems of cyanobacteriae Mycrocystis aeruginosa. The bench allows regulating the luminous intensity and suspension agitation, studying the cells reproduction and conducting the chemical analysis of water. It has been established that algae can be used in a biofilter for reducing low concentrations of total iron in water to the levels which meet state standard (GOST 2874-82 requirements. Algae cells reproduction during water treatment from iron in a biofilter indicates that the number of enzyme systems and their activity increase. The advantage of the algae multienzyme systems use for the water treatment is the fact that they maintain their activity for a long time and do not need any carbonaceous energy substrata.

  20. Prolonged outbreak of clonal MDR Pseudomonas aeruginosa on an intensive care unit: contaminated sinks and contamination of ultra-filtrate bags as possible route of transmission?

    Directory of Open Access Journals (Sweden)

    Florian Salm

    2016-12-01

    -up period, the outbreak strain was detected only once, which indicated that the outbreak had been ceased (incidence 0.75% vs. 0.04%, p < 0.001 Furthermore, the incidence of Pseudonomas aeruginosa overall was significantly decreased (2.5% vs. 1.5%, p < 0.001. Conclusion In ICUs, limiting work processes involving sinks results in reduced multidrug-resistant Pseudomonas aeruginosa rates. ICUs with high rates of Pseudomonas aeruginosa should consider eliminating work processes that involve sinks and potentially splash water in close proximity to patients. Trial registration All data were surveillance based data which were obtained within the German Law on Protection against Infection (“Infektionsschutzgesetz”. Therefore a trial registration was not required.