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Sample records for biofilm voltammetry direct

  1. A new approach for in situ cyclic voltammetry of a microbial fuel cell biofilm without using a potentiostat.

    Science.gov (United States)

    Cheng, Ka Yu; Cord-Ruwisch, Ralf; Ho, Goen

    2009-02-01

    Electrochemically active bacteria in a microbial fuel cell (MFC) usually exist as a biofilm attached to an electrode surface. Conventional cyclic voltammetry using potentiostat is considered as a powerful and reliable method to study electrochemical behavior of MFC biofilm. In this paper, we introduce a new approach to evaluate redox behavior of an electro-active MFC biofilm without using a potentiostat. Analogous to a conventional cyclic voltammetry study, we controlled the biofilm-electrode potential by computer-feedback controlling the external resistance of an operating MFC. In this way, the MFC can still operate as a "fuel cell" without being "interrupted" by an external device (i.e. potentiostat) that normally does not belong to the system. Relationship between current and biofilm-electrode potential was obtained and showed agreement with a potentiostat-controlled method under similar experimental conditions. The method could be added to our technical repertoire for analysis of bacterial mediator involved in the exocellular electron transfer of a MFC-biofilm, and it could potentially serve as a practical process monitoring method for MFC operation. The application of computer-control components should be further explored to facilitate control, diagnosis as well as optimization of MFC processes.

  2. Direct Electrical Current Reduces Bacterial and Yeast Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Maria Ruiz-Ruigomez

    2016-01-01

    Full Text Available New strategies are needed for prevention of biofilm formation. We have previously shown that 24 hr of 2,000 µA of direct current (DC reduces Staphylococcus epidermidis biofilm formation in vitro. Herein, we examined the effect of a lower amount of DC exposure on S. epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Propionibacterium acnes, and Candida albicans biofilm formation. 12 hr of 500 µA DC decreased S. epidermidis, S. aureus, E. coli, and P. aeruginosa biofilm formation on Teflon discs by 2, 1, 1, and 2 log10 cfu/cm2, respectively (p<0.05. Reductions in S. epidermidis, S. aureus, and E. coli biofilm formation were observed with as few as 12 hr of 200 µA DC (2, 2 and 0.4 log10 cfu/cm2, resp.; a 1 log10 cfu/cm2 reduction in P. aeruginosa biofilm formation was observed at 36 hr. 24 hr of 500 µA DC decreased C. albicans biofilm formation on Teflon discs by 2 log10 cfu/cm2. No reduction in P. acnes biofilm formation was observed. 1 and 2 log10 cfu/cm2 reductions in E. coli and S. epidermidis biofilm formation on titanium discs, respectively, were observed with 12 hr of exposure to 500 µA. Electrical current is a potential strategy to reduce biofilm formation on medical biomaterials.

  3. Myoglobin immobilization on electrodeposited nanometer-scale nickel oxide particles and direct voltammetry.

    Science.gov (United States)

    Moghaddam, Abdolmajid Bayandori; Ganjali, Mohammad Reza; Dinarvand, Rassoul; Ahadi, Sara; Saboury, Ali Akbar

    2008-04-01

    Prosperity of information on the reactions of redox-active sites in proteins can be attained by voltammetric studies in which the protein sample is located on a suitable surface. This work reports the presentation of myoglobin/nickel oxide nanoparticles/glassy carbon (Mb/NiO NPs/GC) electrode, ready by electrochemical deposition of the NiO NPs on glassy carbon electrode and myoglobin immobilization on their surfaces by the potential cycling method. Images of electrodeposited NiO NPs on the surface of glassy carbon electrode were obtained by scanning electron microscopy (SEM) and atomic force microscopy (AFM). A pair of well-defined redox peaks for Mb(Fe(III)-Fe(II)) was obtained at the prepared electrode by direct electron transfer between the protein and nanoparticles. Electrochemical parameters of immobilized myoglobin such as formal potential (E(0')), charge transfer coefficient (alpha) and apparent heterogeneous electron transfer rate constant (k(s)) were estimated by cyclic voltammetry and nonlinear regression analysis. Biocatalytic activity was exemplified at the prepared electrode for reduction of hydrogen peroxide.

  4. Femtomolar Detection of Silver Nanoparticles by Flow-Enhanced Direct-Impact Voltammetry at a Microelectrode Array.

    Science.gov (United States)

    Sokolov, Stanislav V; Bartlett, Thomas R; Fair, Peter; Fletcher, Stephen; Compton, Richard G

    2016-09-06

    We report the femtomolar detection of silver (Ag) nanoparticles by direct-impact voltammetry. This is achieved through the use of a random array of microelectrodes (RAM) integrated into a purpose-built flow cell, allowing combined diffusion and convection to the electrode surface. A coupled RAM-flow cell system is implemented and is shown to give reproducible wall-jet type flow characteristics, using potassium ferrocyanide as a molecular redox species. The calibrated flow system is then used to detect and quantitatively size Ag nanoparticles at femtomolar concentrations. Under flow conditions, it is found the nanoparticle impact frequency increases linearly with the volumetric flow rate. The resulting limit of detection is more than 2 orders of magnitude smaller than the previous detection limit for direct-impact voltammetry (900 fM) [J. Ellison et al. Sens. Actuators, B 2014, 200, 47], and is more than 30 times smaller than the previous detection limit for mediated-impact voltammetry (83 fM) [T. M. Alligrant et al. Langmuir 2014, 30, 13462].

  5. Direct Electrochemical Evidence of the Dissociation and Adsorption Behavior of Acetonitrile at Gold Electrodes by Ultrafast Voltammetry

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Ultrafast cyclic voltammetry was used to study the redox behavior of a gold electrode in acetonitrile. The direct electrochemical evidence of the dissociation and adsorption behavior of acetonitrile at gold electrodes was found. It could be stated that two consecutive redox paths are involved, each with a special adsorption state acting as the reaction intermediate. The mean value,obtained of the electron-transfer rate constant of the second path, was 1.3 × 105 s-1 with a standard deviation of 0.24 × 105 s-1.

  6. Pulse Voltammetry.

    Science.gov (United States)

    Osteryoung, Janet

    1983-01-01

    Discusses the nature of pulse voltammetry, indicating that its widespread use arises from good sensitivity and detection limits and from ease of application and low cost. Provides analytical and mechanistic applications of the procedure. (JN)

  7. Direct analysis of palladium in active pharmaceutical ingredients by anodic stripping voltammetry.

    Science.gov (United States)

    Rosolina, Samuel M; Chambers, James Q; Xue, Zi-Ling

    2016-03-31

    Anodic stripping voltammetry, a classical electroanalytical method has been optimized to analyze trace Pd(II) in active pharmaceutical ingredient matrices. The electroanalytical approach with an unmodified glassy carbon electrode was performed in both aqueous and 95% DMSO/5% water (95/5 DMSO/H2O) solutions, without pretreatment such as acid digestion or dry ashing to remove the organics. Limits of detection (LODs) in the presence of caffeine and ketoprofen were determined to be 11 and 9.6 μg g(-1), with a relative standard deviation (RSD) of 5.7% and 2.3%, respectively. This method is simple, highly reproducible, sensitive, and robust. The instrumentation has the potential to be portable and the obviation of sample pretreatment makes it an ideal approach for determining lost catalytic metals in pharmaceutical-related industries. Furthermore, the simultaneous detection of Pd(II) with Cd(II) and Pb(II) in the low μg L(-1) range indicates that this system is capable of simultaneous multi-analyte analysis in a variety of matrices.

  8. Cyclic Voltammetry.

    Science.gov (United States)

    Evans, Dennis H.; And Others

    1983-01-01

    Cyclic voltammetry is a simple experiment that has become popular in chemical research because it can provide useful information about redox reactions in a form which is easily obtained and interpreted. Discusses principles of the method and illustrates its use in the study of four electrode reactions. (Author/JN)

  9. Direct electrodeposition of gold nanotube arrays of rough and porous wall by cyclic voltammetry and its applications of simultaneous determination of ascorbic acid and uric acid.

    Science.gov (United States)

    Yang, Guangming; Li, Ling; Jiang, Jinhe; Yang, Yunhui

    2012-08-01

    Gold nanotube arrays of rough and porous wall has been synthesized by direct electrodeposition with cyclic voltammetry utilizing anodic aluminum oxide template (AAO) and polycarbonate membrane (PC) during short time (only 3 min and 2 min, respectively). The mechanism of the direct electrodeposition of gold nanotube arrays by cyclic voltammetry (CV) has been discussed. The morphological characterizations of the gold nanotube arrays have been investigated by scanning electron microscopy (SEM). A simultaneous determination of ascorbic acid (AA) and uric acid (UA) by differential pulse voltammetry (DPV) was constructed by attaching gold nanotube arrays (using AAO) onto the surface of a glassy carbon electrode (GCE). The electrochemical behavior of AA and UA at this modified electrode has been studied by CV and differential pulse voltammetry (DPV). The sensor offers an excellent response for AA and UA and the linear response range for AA and UA were 1.02×10(-7)-5.23×10(-4) mol L(-1) and 1.43×10(-7)-4.64×10(-4) mol L(-1), the detection limits were 1.12×10(-8) mol L(-1) and 2.24×10(-8) mol L(-1), respectively. This sensor shows good regeneration, stability and selectivity and has been used for the determination of AA and UA in real human urine and serum samples with satisfied results.

  10. Direct measurement and characterization of active photosynthesis zones inside biofuel producing and wastewater remediating microalgal biofilms

    Energy Technology Data Exchange (ETDEWEB)

    Bernstein, Hans C.; Kesaano, Maureen; Moll, Karen; Smith, Terence; Gerlach, Robin; Carlson, Ross; Miller, Charles D.; Peyton, Brent; Cooksey, Keith; Gardner, Robert D.; Sims, Ronald C.

    2014-03-01

    Abstract: Microalgal biofilm based technologies are of keen interest due to their high biomass concentrations and ability to utilize renewable resources, such as light and CO2. While photoautotrophic biofilms have long been used for wastewater remediation applications, biofuel production represents a relatively new and under-represented focus area. However, the direct measurement and characterization of fundamental parameters required for physiological analyses are challenging due to biofilm heterogeneity. This study evaluated oxygenic photosynthesis and biofuel precursor molecule production using a novel rotating algal biofilm reactor (RABR) operated at field- and laboratory-scales for wastewater remediation and biofuel production, respectively. Clear differences in oxygenic-photosynthesis, respiration and biofuel-precursor capacities were observed between the two systems and different conditions based on light and nitrogen availability. Nitrogen depletion was not found to have the same effect on lipid accumulation compared to prior planktonic studies. Physiological characterizations of these microalgal biofilms identify potential areas for future process optimization.

  11. Direct in situ measurement of dissolved zinc in the presence of zinc oxide nanoparticles using anodic stripping voltammetry.

    Science.gov (United States)

    Jiang, Chuanjia; Hsu-Kim, Heileen

    2014-11-01

    The wide use of metal-based nanomaterials such as zinc oxide (ZnO) nanoparticles (NPs) has generated concerns regarding their environmental and health risks. For ZnO NPs, their toxicity in aquatic systems often depends on the release of dissolved zinc species, and the rate of dissolution is influenced by water chemistry, including the presence of zinc-chelating ligands. A challenge, however, remains in quantifying the dissolution of ZnO NPs, particularly for time scales that are short enough to determine rates. This paper reports the application of anodic stripping voltammetry (ASV) with a hanging mercury drop electrode to directly measure the concentration of dissolved zinc in ZnO NP suspensions, without separation of the ZnO NPs from the aqueous phase. The effects of the deposition time and the electrochemical potential scan rate on the ASV measurement were consistent with expectations for dissolved phase measurements. The dissolved zinc concentration measured by ASV ([Zn]ASV) was compared with that measured by inductively coupled plasma mass spectrometry (ICP-MS) after ultracentrifugation ([Zn]ICP-MS), for four types of ZnO NPs with different coatings and primary particle diameters. For small ZnO NPs (4-5 nm), [Zn]ASV was 20% higher than [Zn]ICP-MS, suggesting that these small NPs contributed to the voltammetric measurement. For larger ZnO NPs (approximately 20 nm), [Zn]ASV was (79 ± 19)% of [Zn]ICP-MS, despite the high concentrations of ZnO NPs in suspension. Using ASV, the dissolution of ZnO NPs was studied, with or without Suwannee River Fulvic Acid (SRFA). Although SRFA diminished the ASV stripping current, dissolution of 20 nm ZnO NPs was significantly promoted at high fulvic acid to ZnO NP ratios. The ASV method described in this paper provides a useful tool for studying the dissolution kinetics of ZnO NPs in complex environmental matrices.

  12. Microbial fuel cell based on Klebsiella pneumoniae biofilm

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Lixia [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Guangdong Institute of Eco-Environmental and Soil Sciences, Guangzhou 510650 (China); Zhou, Shungui; Zhuang, Li; Zhang, Jintao; Lu, Na; Deng, Lifang [Guangdong Institute of Eco-Environmental and Soil Sciences, Guangzhou 510650 (China); Li, Weishan [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Key Laboratory of Electrochemical Technology on Energy Storage and Power Generation in Guangdong Universities, Guangzhou 510006 (China)

    2008-10-15

    In this paper we reported a novel microbial fuel cell (MFC) based on Klebsiella pneumoniae (K. pneumoniae) strain L17 biofilm, which can utilize directly starch and glucose to generate electricity. The electrochemical activity of K. pneumoniae and the performance of the MFC were evaluated by cyclic voltammetry, scanning electron microscope (SEM) and polarization curve measurement. The results indicated that an established K. pneumoniae biofilm cells were responsible for the direct electron transfer from fuels to electrode during electricity production. The SEM observation proved the ability of K. pneumoniae to colonize on the electrode surface. This MFC generated power from the direct electrocatalysis by the K. pneumoniae strain L17 biofilm. (author)

  13. Vibrio cholerae utilizes direct sRNA regulation in expression of a biofilm matrix protein.

    Directory of Open Access Journals (Sweden)

    Tianyan Song

    Full Text Available Vibrio cholerae biofilms contain exopolysaccharide and three matrix proteins RbmA, RbmC and Bap1. While much is known about exopolysaccharide regulation, little is known about the mechanisms by which the matrix protein components of biofilms are regulated. VrrA is a conserved, 140-nt sRNA of V. cholerae, whose expression is controlled by sigma factor σE. In this study, we demonstrate that VrrA negatively regulates rbmC translation by pairing to the 5' untranslated region of the rbmC transcript and that this regulation is not stringently dependent on the RNA chaperone protein Hfq. These results point to VrrA as a molecular link between the σE-regulon and biofilm formation in V. cholerae. In addition, VrrA represents the first example of direct regulation of sRNA on biofilm matrix component, by-passing global master regulators.

  14. Development of Enterobacter aerogenes fuel cells: from in situ biohydrogen oxidization to direct electroactive biofilm.

    Science.gov (United States)

    Zhuang, Li; Zhou, Shungui; Yuan, Yong; Liu, Tinglin; Wu, Zhifeng; Cheng, Jiong

    2011-01-01

    This study described an Enterobacter aerogenes-catalyzed microbial fuel cell (MFC) with a carbon-based anode that exhibited a maximum power density of 2.51 W/m(3) in the absence of artificial electron mediators. The MFC was started up rapidly, within hours, and the current generation in the early stage was demonstrated to result from in situ oxidation of biohydrogen produced by E. aerogenes during glucose fermentation. Over periodic replacement of substrate, both planktonic biomass in the culture liquid and hydrogen productivity decreased, while increased power density and coulombic efficiency and decreased internal resistance were unexpectedly observed. Using scanning electron microscopy and cyclic voltammetry, it was found that the enhanced MFC performance was associated with the development of electroactive biofilm on the anodic surface, proposed to involve an acclimation and selection process of E. aerogenes cells under electrochemical tension. The significant advantage of rapid start-up and the ability to develop an electroactive biofilm identifies E. aerogenes as a suitable biocatalyst for MFC applications.

  15. Direct determination of cadmium and lead in pharmaceutical ingredients using anodic stripping voltammetry in aqueous and DMSO/water solutions.

    Science.gov (United States)

    Rosolina, Samuel M; Chambers, James Q; Lee, Carlos W; Xue, Zi-Ling

    2015-09-17

    A new electrochemical method has been developed to detect and quantify the elemental impurities, cadmium(II) (Cd(2+)) and lead(II) (Pb(2+)), either simultaneously or individually in pharmaceutical matrices. The electro-analytical approach, involving the use of anodic stripping voltammetry (ASV) on an unmodified glassy carbon electrode, was performed in both aqueous and in a 95/5 dimethyl sulfoxide (DMSO)/water solutions, without acid digestion or dry ashing to remove organic matrices. Limits of detection (LODs) in the μg L(-1) [or parts per billion (ppb), mass/volume] range were obtained for both heavy metals - in the presence and absence of representative pharmaceutical components. To the best of our knowledge, the work demonstrates the first analysis of heavy metals in DMSO/water solutions through ASV. The strong reproducibility and stability of the sensing platform, as well as obviation of sample pretreatment show the promise of utilizing ASV as a sensitive, robust, and inexpensive alternative to inductively-coupled-plasma (ICP)-based approaches for the analysis of elemental impurities in, e.g., pharmaceutical-related matrices.

  16. Electron transfer mechanism in Shewanella loihica PV-4 biofilms formed at graphite electrode.

    Science.gov (United States)

    Jain, Anand; Zhang, Xiaoming; Pastorella, Gabriele; Connolly, Jack O; Barry, Niamh; Woolley, Robert; Krishnamurthy, Satheesh; Marsili, Enrico

    2012-10-01

    Electron transfer mechanisms in Shewanella loihica PV-4 viable biofilms formed at graphite electrodes were investigated in potentiostat-controlled electrochemical cells poised at oxidative potentials (0.2V vs. Ag/AgCl). Chronoamperometry (CA) showed a repeatable biofilm growth of S. loihica PV-4 on graphite electrode. CA, cyclic voltammetry (CV) and its first derivative shows that both direct electron transfer (DET) mediated electron transfer (MET) mechanism contributes to the overall anodic (oxidation) current. The maximum anodic current density recorded on graphite was 90 μA cm(-2). Fluorescence emission spectra shows increased concentration of quinone derivatives and riboflavin in the cell-free supernatant as the biofilm grows. Differential pulse voltammetry (DPV) show accumulation of riboflavin at the graphite interface, with the increase in incubation period. This is the first study to observe a gradual shift from DET to MET mechanism in viable S. loihica PV-4 biofilms.

  17. Wound biofilms: lessons learned from oral biofilms.

    Science.gov (United States)

    Mancl, Kimberly A; Kirsner, Robert S; Ajdic, Dragana

    2013-01-01

    Biofilms play an important role in the development and pathogenesis of many chronic infections. Oral biofilms, more commonly known as dental plaque, are a primary cause of oral diseases including caries, gingivitis, and periodontitis. Oral biofilms are commonly studied as model biofilm systems as they are easily accessible; thus, biofilm research in oral diseases is advanced with details of biofilm formation and bacterial interactions being well elucidated. In contrast, wound research has relatively recently directed attention to the role biofilms have in chronic wounds. This review discusses the biofilms in periodontal disease and chronic wounds with comparisons focusing on biofilm detection, biofilm formation, the immune response to biofilms, bacterial interaction, and quorum sensing. Current treatment modalities used by both fields and future therapies are also discussed.

  18. Affordable Cyclic Voltammetry

    Science.gov (United States)

    Stewart, Greg; Kuntzleman, Thomas S.; Amend, John R.; Collins, Michael J.

    2009-01-01

    Cyclic voltammetry is an important component of the undergraduate chemical curriculum. Unfortunately, undergraduate students rarely have the opportunity to conduct experiments in cyclic voltammetry owing to the high cost of potentiostats, which are required to control these experiments. By using MicroLab data acquisition interfaces in conjunction…

  19. Direct Comparison of Physical Properties of Bacillus subtilis NCIB 3610 and B-1 Biofilms.

    Science.gov (United States)

    Kesel, Sara; Grumbein, Stefan; Gümperlein, Ina; Tallawi, Marwa; Marel, Anna-Kristina; Lieleg, Oliver; Opitz, Madeleine

    2016-04-01

    Many bacteria form surface-attached communities known as biofilms. Due to the extreme resistance of these bacterial biofilms to antibiotics and mechanical stresses, biofilms are of growing interest not only in microbiology but also in medicine and industry. Previous studies have determined the extracellular polymeric substances present in the matrix of biofilms formed by Bacillus subtilis NCIB 3610. However, studies on the physical properties of biofilms formed by this strain are just emerging. In particular, quantitative data on the contributions of biofilm matrix biopolymers to these physical properties are lacking. Here, we quantitatively investigated three physical properties of B. subtilis NCIB 3610 biofilms: the surface roughness and stiffness and the bulk viscoelasticity of these biofilms. We show how specific biomolecules constituting the biofilm matrix formed by this strain contribute to those biofilm properties. In particular, we demonstrate that the surface roughness and surface elasticity of 1-day-old NCIB 3610 biofilms are strongly affected by the surface layer protein BslA. For a second strain,B. subtilis B-1, which forms biofilms containing mainly γ-polyglutamate, we found significantly different physical biofilm properties that are also differently affected by the commonly used antibacterial agent ethanol. We show that B-1 biofilms are protected from ethanol-induced changes in the biofilm's stiffness and that this protective effect can be transferred to NCIB 3610 biofilms by the sole addition of γ-polyglutamate to growing NCIB 3610 biofilms. Together, our results demonstrate the importance of specific biofilm matrix components for the distinct physical properties of B. subtilis biofilms.

  20. Population dynamics and spatial distribution of microbial species in multispecies biofilms under the action of direct electric current

    Institute of Scientific and Technical Information of China (English)

    CAO Hongbin; LI Xingang; WU Jinchuan; ZHONG Fangli; ZHANG Yi

    2003-01-01

    The metabolism, population dynamics and spatial distribution of nitrifying bacteria and heterotrophs in biofilms under the action of direct electric current were investigated by using the micro-slicing technique. The nitrification rate of nitrifying bacteria was severely inhibited by a current over 10 Am-2 at lower C/N ratios. Compared to heterotrophs, the nitrifying bacteria in the surface biofilms were severely inhibited, resulting in a significant decrease in bacterial density. An increase in current density narrowed the less current-sensitive inner biofilm region, and in addition the density of NO2-oxidizers decreased more significantly than that of NH4-oxidizers in the surface biofilms probably due to electrochemical reactions at the anode. However, the effect of current on both the population dynamics and the spatial distribution of the microbial species was less significant at larger C/N ratios.

  1. Direct Determination of Zinc, Cadmium, Lead, Copper Metal in Tap Water of Delhi (India by Anodic Stripping Voltammetry Technique

    Directory of Open Access Journals (Sweden)

    Raj J.

    2013-04-01

    Full Text Available Salts of Zinc, Cadmium, Lead and Copper are taken incidentally or accidently and has become of great toxicological importance having toxic effect. In the present study direct determination of Zn, Cd, Pb and Cu metal was carried out from tap water of Delhi (India using differential pulse anodic stripping Voltammeter (DPASV at Hanging mercury dropping electrode (HMDE.Determination of Zn, Cd, Pb, Cu was done using Ammonium acetate buffer (pH 4.6 with a sweep rate (scan rate of 59.5 mV/s and pulse amplitude 50mV by HMDE by standard addition method. The solution was stirred during pre-electrolysis at -1150mV (vs. Ag/AgCl for 90 seconds and the potential was scanned from -1150V to +100V (vs..Ag/AgCl. As a result the minimum level of Zn, Cd, Pb, Cu was Zero and the concentration observed in the tap water sample of Delhi (India was determined as 0.174 mg/L-1, 0.001 mg/L-1, 0.002 mg/L-1, 0.011 mg/L-1 respectively.

  2. Voltammetry Method Evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Hoyt, N. [Argonne National Lab. (ANL), Argonne, IL (United States); Pereira, C. [Argonne National Lab. (ANL), Argonne, IL (United States); Willit, J. [Argonne National Lab. (ANL), Argonne, IL (United States); Williamson, M. [Argonne National Lab. (ANL), Argonne, IL (United States)

    2016-07-29

    The purpose of the ANL MPACT Voltammetry project is to evaluate the suitability of previously developed cyclic voltammetry techniques to provide electroanalytical measurements of actinide concentrations in realistic used fuel processing scenarios. The molten salts in these scenarios are very challenging as they include high concentrations of multiple electrochemically active species, thereby creating a variety of complications. Some of the problems that arise therein include issues related to uncompensated resistance, cylindrical diffusion, and alloying of the electrodeposited metals. Improvements to the existing voltammetry technique to account for these issues have been implemented, resulting in good measurements of actinide concentrations across a wide range of adverse conditions.

  3. Recent Advances in Voltammetry.

    Science.gov (United States)

    Batchelor-McAuley, Christopher; Kätelhön, Enno; Barnes, Edward O; Compton, Richard G; Laborda, Eduardo; Molina, Angela

    2015-06-01

    Recent progress in the theory and practice of voltammetry is surveyed and evaluated. The transformation over the last decade of the level of modelling and simulation of experiments has realised major advances such that electrochemical techniques can be fully developed and applied to real chemical problems of distinct complexity. This review focuses on the topic areas of: multistep electrochemical processes, voltammetry in ionic liquids, the development and interpretation of theories of electron transfer (Butler-Volmer and Marcus-Hush), advances in voltammetric pulse techniques, stochastic random walk models of diffusion, the influence of migration under conditions of low support, voltammetry at rough and porous electrodes, and nanoparticle electrochemistry. The review of the latter field encompasses both the study of nanoparticle-modified electrodes, including stripping voltammetry and the new technique of 'nano-impacts'.

  4. Recent Advances in Voltammetry

    Science.gov (United States)

    Batchelor-McAuley, Christopher; Kätelhön, Enno; Barnes, Edward O; Compton, Richard G; Laborda, Eduardo; Molina, Angela

    2015-01-01

    Recent progress in the theory and practice of voltammetry is surveyed and evaluated. The transformation over the last decade of the level of modelling and simulation of experiments has realised major advances such that electrochemical techniques can be fully developed and applied to real chemical problems of distinct complexity. This review focuses on the topic areas of: multistep electrochemical processes, voltammetry in ionic liquids, the development and interpretation of theories of electron transfer (Butler–Volmer and Marcus–Hush), advances in voltammetric pulse techniques, stochastic random walk models of diffusion, the influence of migration under conditions of low support, voltammetry at rough and porous electrodes, and nanoparticle electrochemistry. The review of the latter field encompasses both the study of nanoparticle-modified electrodes, including stripping voltammetry and the new technique of ‘nano-impacts’. PMID:26246984

  5. Attributes of direct current aperiodic and alternating current harmonic components derived from large amplitude Fourier transformed voltammetry under microfluidic control in a channel electrode.

    Science.gov (United States)

    Matthews, Sinéad M; Shiddiky, Muhammad J A; Yunus, Kamran; Elton, Darrell M; Duffy, Noel W; Gu, Yunfeng; Fisher, Adrian C; Bond, Alan M

    2012-08-07

    The flow rate dependencies of the aperiodic direct current (dc) and fundamental to eighth alternating current (ac) harmonic components derived from large-amplitude Fourier transformed ac (FT-ac) voltammetry have been evaluated in a microfluidic flow cell containing a 25 μm gold microband electrode. For the oxidation of ferrocenemethanol ([FcMeOH]/[FcMeOH](+) process) in aqueous 0.1 M KNO(3) electrolyte, standard "Levich-like" dc behavior is observed for the aperiodic dc component, which enables the diffusion coefficient for FcMeOH to be obtained. In experimental studies, the first and second ac harmonic components contain contributions from the double layer capacitance current, thereby allowing details of the non-Faradaic current to be established. In contrast, the higher order harmonics and dc aperiodic component are essentially devoid of double layer capacitance contributions allowing the faradaic current dependence on flow rate to be studied. Significantly, flow rate independent data conforming to linear diffusion controlled theory are found in the sixth and higher ac harmonics at a frequency of 15 Hz and for all ac harmonics at a frequency of ≥ 90 Hz. Analysis of FT-ac voltammograms by theory based on stationary microband or planar electrode configurations confirms that stationary microband and planar electrode configurations and experimental data all converge for the higher order harmonics and establishes that the electrode kinetics are very fast (≥1 cms(-1)). The ability to locate, from a single experiment, a dc Faradaic component displaying Levich behavior, fundamental and second harmonics that contain details of the double layer capacitance, and Faradaic ac higher order harmonic currents that are devoid of capacitance, independent of the volume flow rate and also conform closely to mass transport by planar diffusion, provides enhanced flexibility in mass transport and electrode kinetic analysis and in understanding the performance of hydrodynamic

  6. Direct determination of tellurium and its redox speciation at the low nanogram level in natural waters by catalytic cathodic stripping voltammetry.

    Science.gov (United States)

    Biver, Marc; Quentel, François; Filella, Montserrat

    2015-11-01

    Tellurium is one of the elements recently identified as technologically critical and is becoming a new emergent contaminant. No reliable method exists for its determination in environmental samples such as natural waters. This gap is filled by the method described here; it allows the rapid detection of trace concentrations of Te(IV) and Te(VI) in surface waters by differential pulse cathodic stripping voltammetry. It is based on the proton reduction catalysed by the absorption of Te(IV) on the mercury electrode. Under our conditions (0.1 mol L(-1) HCl) a detection limit of about 5 ng L(-1) for a deposition time of 300 s is achieved. Organic matter does not represent a problem at low concentrations; higher concentrations are eliminated by adsorptive purification. Tellurium occurs primarily as Te(IV) and Te(VI) in natural waters. Thus, determining total Te requires the reduction of Te(VI) that it is not electroactive. A number of reduction procedures have been carefully evaluated and a method based on the addition of TiCl3 to the acidified samples has been proven to reduce Te(VI) at the trace level to Te(IV) reliably and quantitatively. Therefore, the procedure described allows the direct determination of total Te and its redox speciation. It is flexible, reliable and cost effective compared to any possible alternative method based on the common preconcentration-ICPMS approach. It is readily implementable as a routine method and can be deployed in the field with relative ease.

  7. Bad to the Bone: On In Vitro and Ex Vivo Microbial Biofilm Ability to Directly Destroy Colonized Bone Surfaces without Participation of Host Immunity or Osteoclastogenesis

    Science.gov (United States)

    Junka, Adam; Szymczyk, Patrycja; Ziółkowski, Grzegorz; Karuga-Kuzniewska, Ewa; Smutnicka, Danuta; Bil-Lula, Iwona; Bartoszewicz, Marzenna; Mahabady, Susan; Sedghizadeh, Parish Paymon

    2017-01-01

    Bone infections are a significant public health burden associated with morbidity and mortality in patients. Microbial biofilm pathogens are the causative agents in chronic osteomyelitis. Research on the pathogenesis of osteomyelitis has focused on indirect bone destruction by host immune cells and cytokines secondary to microbial insult. Direct bone resorption by biofilm pathogens has not yet been seriously considered. In this study, common osteomyelitis pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Streptococcus mutans) were grown as biofilms in multiple in vitro and ex vivo experiments to analyze quantitative and qualitative aspects of bone destruction during infection. Pathogens were grown as single or mixed species biofilms on the following substrates: hydroxyapatite, rat jawbone, or polystyrene wells, and in various media. Biofilm growth was evaluated by scanning electron microscopy and pH levels were monitored over time. Histomorphologic and quantitative effects of biofilms on tested substrates were analyzed by microcomputed tomography and quantitative cultures. All tested biofilms demonstrated significant damage to bone. Scanning electron microscopy indicated that all strains formed mature biofilms within 7 days on all substrate surfaces regardless of media. Experimental conditions impacted pH levels, although this had no impact on biofilm growth or bone destruction. Presence of biofilm led to bone dissolution with a decrease of total volume by 20.17±2.93% upon microcomputed tomography analysis, which was statistically significant as compared to controls (p <0.05, ANOVA). Quantitative cultures indicated that media and substrate did not impact biofilm formation (Kruskall-Wallis test, post-hoc Dunne’s test; p <0.05). Overall, these results indicate that biofilms associated with osteomyelitis have the ability to directly resorb bone. These findings should lead to a more complete understanding of the etiopathogenesis of

  8. Direct electric current treatment under physiologic saline conditions kills Staphylococcus epidermidis biofilms via electrolytic generation of hypochlorous acid.

    Directory of Open Access Journals (Sweden)

    Elizabeth L Sandvik

    Full Text Available The purpose of this study was to investigate the mechanism by which a direct electrical current reduced the viability of Staphylococcus epidermidis biofilms in conjunction with ciprofloxacin at physiologic saline conditions meant to approximate those in an infected artificial joint. Biofilms grown in CDC biofilm reactors were exposed to current for 24 hours in 1/10(th strength tryptic soy broth containing 9 g/L total NaCl. Dose-dependent log reductions up to 6.7 log(10 CFU/cm(2 were observed with the application of direct current at all four levels (0.7 to 1.8 mA/cm(2 both in the presence and absence of ciprofloxacin. There were no significant differences in log reductions for wells with ciprofloxacin compared to those without at the same current levels. When current exposures were repeated without biofilm or organics in the medium, significant generation of free chlorine was measured. Free chlorine doses equivalent to the 24 hour endpoint concentration for each current level were shown to mimic killing achieved by current application. Current exposure (1.8 mA/cm(2 in medium lacking chloride and amended with sulfate, nitrate, or phosphate as alternative electrolytes produced diminished kills of 3, 2, and 0 log reduction, respectively. Direct current also killed Pseudomonas aeruginosa biofilms when NaCl was present. Together these results indicate that electrolysis reactions generating hypochlorous acid from chloride are likely a main contributor to the efficacy of direct current application. A physiologically relevant NaCl concentration is thus a critical parameter in experimental design if direct current is to be investigated for in vivo medical applications.

  9. Direct electric current treatment under physiologic saline conditions kills Staphylococcus epidermidis biofilms via electrolytic generation of hypochlorous acid.

    Science.gov (United States)

    Sandvik, Elizabeth L; McLeod, Bruce R; Parker, Albert E; Stewart, Philip S

    2013-01-01

    The purpose of this study was to investigate the mechanism by which a direct electrical current reduced the viability of Staphylococcus epidermidis biofilms in conjunction with ciprofloxacin at physiologic saline conditions meant to approximate those in an infected artificial joint. Biofilms grown in CDC biofilm reactors were exposed to current for 24 hours in 1/10(th) strength tryptic soy broth containing 9 g/L total NaCl. Dose-dependent log reductions up to 6.7 log(10) CFU/cm(2) were observed with the application of direct current at all four levels (0.7 to 1.8 mA/cm(2)) both in the presence and absence of ciprofloxacin. There were no significant differences in log reductions for wells with ciprofloxacin compared to those without at the same current levels. When current exposures were repeated without biofilm or organics in the medium, significant generation of free chlorine was measured. Free chlorine doses equivalent to the 24 hour endpoint concentration for each current level were shown to mimic killing achieved by current application. Current exposure (1.8 mA/cm(2)) in medium lacking chloride and amended with sulfate, nitrate, or phosphate as alternative electrolytes produced diminished kills of 3, 2, and 0 log reduction, respectively. Direct current also killed Pseudomonas aeruginosa biofilms when NaCl was present. Together these results indicate that electrolysis reactions generating hypochlorous acid from chloride are likely a main contributor to the efficacy of direct current application. A physiologically relevant NaCl concentration is thus a critical parameter in experimental design if direct current is to be investigated for in vivo medical applications.

  10. Cyclic Voltammetry Experiment.

    Science.gov (United States)

    Van Benschoten, James J.; And Others

    1983-01-01

    Describes a three-part experiment designed to introduce cyclic voltammetry to graduate/undergraduate students. Part 1 demonstrates formal reduction potential, redox electron transfer, diffusion coefficient, and electrochemical reversibility. Part 2 investigates electrochemical behavior of acetaminophen. Part 3 examines such experimental variables…

  11. Voltammetry of Medical Biomaterials

    OpenAIRE

    Gulaboski, Rubin; Markovski, Velo

    2015-01-01

    The use of biomaterials in the medicine, dentistry and pharmacy represents probably a major breakthrough in tackling many diseases or disabilities in the last 50 years. We refer to varios techniques that are used for the characterization of the structure and the composition of the biomaterials. Voltammetry is an electrochemical technique that helps mainly in understanding the redox properties of various biomaterials containing some suitable redox centers in their structure. We give in this le...

  12. Rapid direct methods for enumeration of specific, active bacteria in water and biofilms

    Science.gov (United States)

    McFeters, G. A.; Pyle, B. H.; Lisle, J. T.; Broadaway, S. C.

    1999-01-01

    Conventional methods for detecting indicator and pathogenic bacteria in water may underestimate the actual population due to sublethal environmental injury, inability of the target bacteria to take up nutrients and other physiological factors which reduce bacterial culturability. Rapid and direct methods are needed to more accurately detect and enumerate active bacteria. Such a methodological advance would provide greater sensitivity in assessing the microbiological safety of water and food. The principle goal of this presentation is to describe novel approaches we have formulated for the rapid and simultaneous detection of bacteria plus the determination of their physiological activity in water and other environmental samples. The present version of our method involves the concentration of organisms by membrane filtration or immunomagnetic separation and combines an intracellular fluorochrome (CTC) for assessment of respiratory activity plus fluorescent-labelled antibody detection of specific bacteria. This approach has also been successfully used to demonstrate spatial and temporal heterogeneities of physiological activities in biofilms when coupled with cryosectioning. Candidate physiological stains include those capable of determining respiratory activity, membrane potential, membrane integrity, growth rate and cellular enzymatic activities. Results obtained thus far indicate that immunomagnetic separation can provide a high degree of sensitivity in the recovery of seeded target bacteria (Escherichia coli O157:H7) in water and hamburger. The captured and stained target bacteria are then enumerated by either conventional fluorescence microscopy or ChemScan(R), a new instrument that is very sensitive and rapid. The ChemScan(R) laser scanning instrument (Chemunex, Paris, France) provides the detection of individual fluorescently labelled bacterial cells using three emission channels in less than 5 min. A high degree of correlation has been demonstrated between

  13. Pulse Voltammetry in Single Cells Using Platinum Microelectrodes

    Science.gov (United States)

    1991-11-22

    ring electrodes [18) in a solution of 1.OxlO’ M H2PtCl6 and 0.5 M H2SO4 and reducing platinum at 0.0 V vs SSCE for a desired deposition time. Cyclic ...E. and the range for Ed in multiple pulse voltammetry can be chosen from examination of voltammograms obtained by cyclic voltammetry or lin-ir sweep... voltametry [3,13]. As pointed out by Sinru et al. [14) the potential and time of each pulse has a direct effect on the nature of the voltammetry

  14. Biofilm Risks

    DEFF Research Database (Denmark)

    Wirtanen, Gun Linnea; Salo, Satu

    2016-01-01

    This chapter on biofilm risks deals with biofilm formation of pathogenic microbes, sampling and detection methods, biofilm removal, and prevention of biofilm formation. Several common pathogens produce sticky and/or slimy structures in which the cells are embedded, that is, biofilms, on various s...

  15. Layer by layer assembly of catalase and amine-terminated ionic liquid onto titanium nitride nanoparticles modified glassy carbon electrode: Study of direct voltammetry and bioelectrocatalytic activity

    Energy Technology Data Exchange (ETDEWEB)

    Saadati, Shagayegh [Department of Chemistry, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of); Salimi, Abdollah, E-mail: absalimi@uok.ac.ir [Department of Chemistry, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of); Research Center for Nanotechnology, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of); Hallaj, Rahman; Rostami, Amin [Department of Chemistry, University of Kurdistan, P.O. Box 416, Sanandaj (Iran, Islamic Republic of)

    2012-11-13

    Highlights: Black-Right-Pointing-Pointer Catalase and amine-terminated ionic liquid were immobilized to GC/TiNnp with LBL assembly method. Black-Right-Pointing-Pointer First a thin layer of NH{sub 2}-IL is covalently attached to GC/TiNnp electrode using electro-oxidation. Black-Right-Pointing-Pointer With alternative assemble of IL and catalase with positive and negative charged, multilayer was formed. Black-Right-Pointing-Pointer Immobilized catalase shows excellent electrocatalytic activity toward H{sub 2}O{sub 2} reduction. Black-Right-Pointing-Pointer Biosensor response is directly correlated to the number of bilayers. - Abstract: A novel, simple and facile layer by layer (LBL) approach is used for modification of glassy carbon (GC) electrode with multilayer of catalase and nanocomposite containing 1-(3-Aminopropyl)-3-methylimidazolium bromide (amine terminated ionic liquid (NH{sub 2}-IL)) and titanium nitride nanoparticles (TiNnp). First a thin layer of NH{sub 2}-IL is covalently attached to GC/TiNnp electrode using electro-oxidation method. Then, with alternative self assemble positively charged NH{sub 2}-IL and negatively charged catalase a sensitive H{sub 2}O{sub 2} biosensor is constructed, whose response is directly correlated to the number of bilayers. The surface coverage of active catalase per bilayer, heterogeneous electron transfer rate constant (k{sub s}) and Michaelis-Menten constant (K{sub M}) of immobilized catalase were 3.32 Multiplication-Sign 10{sup -12} mol cm{sup -2}, 5.28 s{sup -1} and 1.1 mM, respectively. The biosensor shows good stability, high reproducibility, long life-time, and fast amperometric response with the high sensitivity of 380 {mu}A mM{sup -1} cm{sup -2} and low detection limit of 100 nM at concentration range up to 2.1 mM.

  16. Aptasensors Based on Stripping Voltammetry

    Directory of Open Access Journals (Sweden)

    Wenjing Qi

    2016-07-01

    Full Text Available Aptasensors based on stripping voltammetry exhibit several advantages, such as high sensitivity and multi-target detection from stripping voltammetric technology, and high selectivity from the specific binding of apamers with targets. This review comprehensively discusses the recent accomplishments in signal amplification strategies based on nanomaterials, such as metal nanoparticles, semiconductor nanoparticles, and nanocomposite materials, which are detected by stripping voltammetry after suitable dissolution. Focus will be put in discussing multiple amplification strategies that are widely applied in aptasensors for small biomolecules, proteins, disease markers, and cancer cells.

  17. Combating biofilms

    DEFF Research Database (Denmark)

    Yang, Liang; Liu, Yang; Wu, Hong;

    2012-01-01

    Biofilms are complex microbial communities consisting of microcolonies embedded in a matrix of self-produced polymer substances. Biofilm cells show much greater resistance to environmental challenges including antimicrobial agents than their free-living counterparts. The biofilm mode of life...... is believed to significantly contribute to successful microbial survival in hostile environments. Conventional treatment, disinfection and cleaning strategies do not proficiently deal with biofilm-related problems, such as persistent infections and contamination of food production facilities. In this review......, strategies to control biofilms are discussed, including those of inhibition of microbial attachment, interference of biofilm structure development and differentiation, killing of biofilm cells and induction of biofilm dispersion....

  18. Cyclic voltammetry of supported BLMs

    Science.gov (United States)

    Murgasova, Renata; Sabo, Jan; Ottova, Angelica L.; Tien, H. T.

    1996-06-01

    The transfer of an electron across a bilayer lipid membrane (BLM) is one of the BLMs most exciting processes. A number of well known electron mediators have been investigated using the method of cyclic voltammetry on a Teflon coated platinum wire, the tip of which has been modified by a self-assembled bilayer lipid membrane (s-BLM). The electrical capacitance of the s-BLM system was measured as a function of frequency. The results are discussed in terms of electron transfer and redox reactions.

  19. Manipulation of Biofilm Microbial Ecology

    Energy Technology Data Exchange (ETDEWEB)

    White, D.C.; Palmer, R.J., Jr.; Zinn, M.; Smith, C.A.; Burkhalter, R.; Macnaughton, S.J.; Whitaker, K.W.; Kirkegaard, R.D.

    1998-08-15

    The biofilm mode of growth provides such significant advantages to the members of the consortium that most organisms in important habitats are found in biofilms. The study of factors that allow manipulation of biofilm microbes in the biofilm growth state requires that reproducible biofilms be generated. The most effective monitoring of biofilm formation, succession and desaturation is with on-line monitoring of microbial biofilms with flowcell for direct observation. The biofilm growth state incorporates a second important factor, the heterogeneity in distribution in time and space of the component members of the biofilm consortium. This heterogeneity is reflected not only in the cellular distribution but in the metabolic activity within a population of cells. Activity and cellular distribution can be mapped in four dimensions with confocal microscopy, and function can be ascertained by genetically manipulated reporter functions for specific genes or by vital stains. The methodology for understanding the microbial ecology of biofilms is now much more readily available and the capacity to manipulate biofilms is becoming an important feature of biotechnology.

  20. Permeabilizing biofilms

    Science.gov (United States)

    Soukos, Nikolaos S.; Lee, Shun; Doukas,; Apostolos G.

    2008-02-19

    Methods for permeabilizing biofilms using stress waves are described. The methods involve applying one or more stress waves to a biofilm, e.g., on a surface of a device or food item, or on a tissue surface in a patient, and then inducing stress waves to create transient increases in the permeability of the biofilm. The increased permeability facilitates delivery of compounds, such as antimicrobial or therapeutic agents into and through the biofilm.

  1. Inactivation of Candida Strains in Planktonic and Biofilm Forms Using a Direct Current, Atmospheric-Pressure Cold Plasma Micro-Jet

    Science.gov (United States)

    Zhu, Wei-Dong; Sun, Peng; Sun, Yi; Yu, Shuang; Wu, Haiyan; Liu, Wei; Zhang, Jue; Fang, Jing

    A direct-current, atmospheric-pressure, He/O2 (2%) cold plasma ­microjet is applied to Candida species (C. glabrata, C. albicansand C. krusei). Effective inactivation is achieved both in air and in water within 5 min of plasma treatment. Same plasma treatment also successfully inactivated candida biofilms on Petri dish. The inactivation was verified by cell viability test (XTT assay). Severe deformation of Candida biofilms after the plasma treatment was observed through scanning electron microscope (SEM). Optical emission spectroscopy shows strong atomic oxygen emission at 777 nm. Hydroxyl radical (•OH), superoxide anion radical (•O2-) and singlet molecular oxygen (1O2) are detected by electron spin resonance (ESR) spectroscopy. The sessile minimal inhibitory concentrations (SMICs) of fluconazole, amphotericin B, and caspofungin against the Candida spp. biofilms were decreased to 2-6 fold dilutions in plasma microjet treated group in comparison with the controls. This novel approach may become a new tool for the treatment of clinical dermatosis

  2. Beneficial biofilms

    Directory of Open Access Journals (Sweden)

    Sara R Robertson

    2015-10-01

    Full Text Available Surface-adherent biofilm growth is a common trait of bacteria and other microorganisms in nature. Within biofilms, organisms are present in high density and are enmeshed in an organic matrix containing polysaccharides and other molecules. The close proximity of organisms within biofilms facilitates microbial interactions and signaling, including many metabolic processes in which consortia rather than individual organisms participate. Biofilm growth also enables microorganisms to withstand chemical and biological stresses. Here, we review some current literature and document representative beneficial aspects of biofilms using examples from wastewater treatment, microbial fuel cells, biological repair (biocementation of stonework, and biofilm protection against Candida albicans infections. Finally, we address a chemical ecology strategy whereby desired microbial succession and beneficial biofilm formation can be encouraged via manipulation of culture conditions and bacterial signaling.

  3. Biofilm roughness determines Cryptosporidium parvum retention in environmental biofilms.

    Science.gov (United States)

    DiCesare, E A Wolyniak; Hargreaves, B R; Jellison, K L

    2012-06-01

    The genus Cryptosporidium is a group of waterborne protozoan parasites that have been implicated in significant outbreaks of gastrointestinal infections throughout the world. Biofilms trap these pathogens and can contaminate water supplies through subsequent release. Biofilm microbial assemblages were collected seasonally from three streams in eastern Pennsylvania and used to grow biofilms in laboratory microcosms. Daily oocyst counts in the influx and efflux flow allowed the calculation of daily oocyst retention in the biofilm. Following the removal of oocysts from the influx water, oocyst attachment to the biofilm declined to an equilibrium state within 5 days that was sustained for at least 25 days. Varying the oocyst loading rate for the system showed that biofilm retention could be saturated, suggesting that discrete binding sites determined the maximum number of oocysts retained. Oocyst retention varied seasonally but was consistent across all three sites; however, seasonal oocyst retention was not consistent across years at the same site. No correlation between oocyst attachment and any measured water quality parameter was found. However, oocyst retention was strongly correlated with biofilm surface roughness and roughness varied among seasons and across years. We hypothesize that biofilm roughness and oocyst retention are dependent on environmentally driven changes in the biofilm community rather than directly on water quality conditions. It is important to understand oocyst transport dynamics to reduce risks of human infection. Better understanding of factors controlling biofilm retention of oocysts should improve our understanding of oocyst transport at different scales.

  4. Understanding Biofilms in Chronic Sinusitis.

    Science.gov (United States)

    Tajudeen, Bobby A; Schwartz, Joseph S; Palmer, James N

    2016-02-01

    Chronic sinusitis is a burdensome disease that has substantial individual and societal impact. Although great advances in medical and surgical therapies have been made, some patients continue to have recalcitrant infections. Microbial biofilms have been implicated as a cause of recalcitrant chronic sinusitis, and recent studies have tried to better understand the pathogenesis of chronic sinusitis as it relates to microbial biofilms. Here, we provide an overview of biofilms in chronic sinusitis with emphasis on pathogenesis, treatment, and future directions. In addition, recent evidence is presented, elucidating the role of bitter taste receptors as a possible key factor leading to biofilm formation.

  5. Electrochemical biofilm control: a review.

    Science.gov (United States)

    Sultana, Sujala T; Babauta, Jerome T; Beyenal, Haluk

    2015-01-01

    One of the methods of controlling biofilms that has widely been discussed in the literature is to apply a potential or electrical current to a metal surface on which the biofilm is growing. Although electrochemical biofilm control has been studied for decades, the literature is often conflicting, as is detailed in this review. The goals of this review are: (1) to present the current status of knowledge regarding electrochemical biofilm control; (2) to establish a basis for a fundamental definition of electrochemical biofilm control and requirements for studying it; (3) to discuss current proposed mechanisms; and (4) to introduce future directions in the field. It is expected that the review will provide researchers with guidelines on comparing datasets across the literature and generating comparable datasets. The authors believe that, with the correct design, electrochemical biofilm control has great potential for industrial use.

  6. Current understanding of multi-species biofilms

    DEFF Research Database (Denmark)

    Yang, Liang; Liu, Yang; Wu, Hong

    2011-01-01

    Direct observation of a wide range of natural microorganisms has revealed the fact that the majority of microbes persist as surface-attached communities surrounded by matrix materials, called biofilms. Biofilms can be formed by a single bacterial strain. However, most natural biofilms are actuall...

  7. Biofilm Infections

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Jensen, Peter Østrup; Moser, Claus Ernst

    A still increasing interest and emphasis on the sessile bacterial lifestyle biofilms has been seen since it was realized that the vast majority of the total microbial biomass exists as biofilms. Aggregation of bacteria was first described by Leeuwenhoek in 1677, but only recently recognized...... as being important in chronic infection. In 1993 the American Society for Microbiology (ASM) recognized that the biofilm mode of growth was relevant to microbiology. This book covers both the evidence for biofilms in many chronic bacterial infections as well as the problems facing these infections...... such as diagnostics, pathogenesis, treatment regimes and in vitro and in vivo models for studying biofilms. This is the first scientific book on biofilm infections, chapters written by the world leading scientist and clinicians. The intended audience of this book is scientists, teachers at university level as well...

  8. Evaluation and Selection of Bacillus Species Based on Enzyme Production, Antimicrobial Activity, and Biofilm Synthesis as Direct-Fed Microbial Candidates for Poultry

    Science.gov (United States)

    Latorre, Juan D.; Hernandez-Velasco, Xochitl; Wolfenden, Ross E.; Vicente, Jose L.; Wolfenden, Amanda D.; Menconi, Anita; Bielke, Lisa R.; Hargis, Billy M.; Tellez, Guillermo

    2016-01-01

    Social concern about misuse of antibiotics as growth promoters (AGP) and generation of multidrug-resistant bacteria have restricted the dietary inclusion of antibiotics in livestock feed in several countries. Direct-fed microbials (DFM) are one of the multiple alternatives commonly evaluated as substitutes of AGP. Sporeformer bacteria from the genus Bacillus have been extensively investigated because of their extraordinary properties to form highly resistant endospores, produce antimicrobial compounds, and synthesize different exogenous enzymes. The purpose of the present study was to evaluate and select Bacillus spp. from environmental and poultry sources as DFM candidates, considering their enzyme production profile, biofilm synthesis capacity, and pathogen-inhibition activity. Thirty-one Bacillus isolates were screened for in vitro relative enzyme activity of amylase, protease, lipase, and phytase using a selective media for each enzyme, with 3/31 strains selected as superior enzyme producers. These three isolates were identified as Bacillus subtilis (1/3), and Bacillus amyloliquefaciens (2/3), based on biochemical tests and 16S rRNA sequence analysis. For evaluation of biofilm synthesis, the generation of an adherent crystal violet-stained ring was determined in polypropylene tubes, resulting in 11/31 strains showing a strong biofilm formation. Moreover, all Bacillus strains were evaluated for growth inhibition activity against Salmonella enterica serovar Enteritidis (26/31), Escherichia coli (28/31), and Clostridioides difficile (29/31). Additionally, in previous in vitro and in vivo studies, these selected Bacillus strains have shown to be resistant to different biochemical conditions of the gastrointestinal tract of poultry. Results of the present study suggest that the selection and consumption of Bacillus-DFM, producing a variable set of enzymes and antimicrobial compounds, may contribute to enhanced performance through improving nutrient digestibility

  9. Evaluation and selection of Bacillus species based on enzyme production, antimicrobial activity and biofilm synthesis as direct-fed microbials candidates for poultry

    Directory of Open Access Journals (Sweden)

    Juan D Latorre

    2016-10-01

    Full Text Available Social concern about misuse of antibiotics as growth promoters (AGP and generation of multidrug-resistant bacteria have restricted the dietary inclusion of antibiotics in livestock feed in several countries. Direct-fed microbials (DFM are one of the multiple alternatives commonly evaluated as substitutes of AGP. Sporeformer bacteria from the genus Bacillus have been extensively investigated because of their extraordinary properties to form highly-resistant endospores, production of antimicrobial compounds and synthesize different exogenous enzymes. The purpose of the present study was to evaluate and select Bacillus spp. from environmental and poultry sources as DFM candidates, considering their enzyme production profile, biofilm synthesis capacity and pathogen-inhibition activity. Thirty one Bacillus isolates were screened for in vitro relative enzyme activity of amylase, protease, lipase and phytase using a selective media for each enzyme, with 3/31 strains selected as superior enzyme producers. These three isolates were identified as B. subtilis (1/3, and B. amyloliquefaciens (2/3 based on biochemical tests and 16S rRNA sequence analysis. For evaluation of biofilm synthesis, the generation of an adherent crystal violet-stained ring was determined in polypropylene tubes, resulting in 11/31 strains showing a strong biofilm formation. Moreover, all Bacillus strains were evaluated for growth inhibition activity against Salmonella enterica serovar Enteritidis (26/31, Escherichia coli (28/31 and Clostridioides difficile (29/31. Additionally, in previous in vitro and in vivo studies, these selected Bacillus strains have shown to be resistant to different biochemical conditions of the gastrointestinal tract of poultry. Results of the present study suggest that the selection and consumption of Bacillus-DFM, producing a variable set of enzymes and antimicrobial compounds may contribute to enhanced performance through improving nutrient digestibility

  10. Salmonella biofilms

    NARCIS (Netherlands)

    Castelijn, G.A.A.

    2013-01-01

    Biofilm formation by Salmonellaspp. is a problem in the food industry, since biofilms may act as a persistent source of product contamination. Therefore the aim of this study was to obtain more insight in the processes involved and the factors contributing to Salmonellabiofilm formation. A collectio

  11. Vaccines directed against microorganisms or their products present during biofilm lifestyle: can we make a translation as a broad biological model to tuberculosis?

    Directory of Open Access Journals (Sweden)

    Mario Alberto eFlores-Valdez

    2016-01-01

    Full Text Available Tuberculosis (TB remains as a global public health problem. In recent years, experimental evidence suggesting the relevance of in vitro pellicle (a type of biofilm formed at the air-liquid interface production as a phenotype mimicking aspects found by M. tuberculosis-complex bacteria during in vivo infection has started to accumulate. There are still opportunities for better diagnostic tools, therapeutic molecules as well as new vaccine candidates to assist in TB control programs worldwide and particularly in less developed nations. Regarding vaccines, despite the availability of a live, attenuated strain (M. bovis BCG since almost a century ago, its variable efficacy and lack of protection against pulmonary and latent disease has prompted basic and applied research leading to preclinical and clinical evaluation of up to 15 new candidates. In this work, I present examples of vaccines based on whole cells grown as biofilms, or specific proteins expressed under such condition, and the effect they have shown in relevant animal models or directly in the natural host. I also discuss why it might be worthwhile to explore these approaches, for constructing and developing new vaccine candidates for testing their efficacy against TB.

  12. Biofilm development

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2015-01-01

    During the past decade we have gained much knowledge about the molecular mechanisms that are involved in initiation and termination of biofilm formation. In many bacteria, these processes appear to occur in response to specific environmental cues and result in, respectively, induction or terminat......During the past decade we have gained much knowledge about the molecular mechanisms that are involved in initiation and termination of biofilm formation. In many bacteria, these processes appear to occur in response to specific environmental cues and result in, respectively, induction...... or termination of biofilm matrix production via the second messenger molecule c-di-GMP. In between initiation and termination of biofilm formation we have defined specific biofilm stages, but the currently available evidence suggests that these transitions are mainly governed by adaptive responses......, and not by specific genetic programs. It appears that biofilm formation can occur through multiple pathways and that the spatial structure of the biofilms is species dependent as well as dependent on environmental conditions. Bacterial subpopulations, e.g., motile and nonmotile subpopulations, can develop...

  13. MATHEMATICAL MODELING OF EXTRACELLULAR ELECTRON TRANSFER IN BIOFILMS

    Energy Technology Data Exchange (ETDEWEB)

    Renslow, Ryan S.; Babauta, Jerome T.; Kuprat, Andrew P.; Schenk, Jim; Ivory, Cornelius; Fredrickson, Jim K.; Beyenal, Haluk

    2015-09-12

    Electrochemically active biofilms have a unique form of respiration in which they utilize solid external materials as terminal electron acceptors for their metabolism. Currently, two primary mechanisms have been identified for long-range extracellular electron transfer (EET): a diffusion- and a conduction-based mechanism. Evidence in the literature suggests that some biofilms, particularly Shewanella oneidensis, produce the requisite components for both mechanisms. In this study, a generic model is presented that incorporates the diffusion- and the conduction-based mechanisms and allows electrochemically active biofilms to utilize both simultaneously. The model was applied to S. oneidensis and Geobacter sulfurreducens biofilms using experimentally generated data found in the literature. Our simulation results show that 1) biofilms having both mechanisms available, especially if they can interact, may have a metabolic advantage over biofilms that can use only a single mechanism; 2) the thickness of G. sulfurreducens biofilms is likely not limited by conductivity; 3) accurate intrabiofilm diffusion coefficient values are critical for current generation predictions; and 4) the local biofilm potential and redox potential are two distinct parameters and cannot be assumed to have identical values. Finally, we determined that simulated cyclic and squarewave voltammetry based on our model are currently not capable of determining the specific percentages of extracellular electron transfer mechanisms in a biofilm. The developed model will be a critical tool for designing experiments to explain EET mechanisms.

  14. Ultrafast cyclic voltammetry with asymmetrical potential scan

    Institute of Scientific and Technical Information of China (English)

    Zhi Yong Guo; Xiang Qin Lin

    2008-01-01

    Based on the perfect ohmic drop compensation by online electronic positive feedback, ultrafast cyclic voltammetry withasymmetrical potential scan is achieved for the first time, with the reduction of anthracene acting as the test system. Compared withthe traditional cyclic voltammetry utilizing symmetrical triangular waveform as the excitation one, the new method allows a simplerapproach to mechanistic analysis of ultrafast chemical reactions coupled with a charge transfer. And perhaps more important, it alsoprovides a way to eliminate the interference of the adsorbed product in dynamic monitoring.

  15. Biofilm-specific extracellular matrix proteins of nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Wu, Siva; Baum, Marc M; Kerwin, James; Guerrero, Debbie; Webster, Simon; Schaudinn, Christoph; VanderVelde, David; Webster, Paul

    2014-12-01

    Nontypeable Haemophilus influenzae (NTHi), a human respiratory tract pathogen, can form colony biofilms in vitro. Bacterial cells and the amorphous extracellular matrix (ECM) constituting the biofilm can be separated using sonication. The ECM from 24- and 96-h NTHi biofilms contained polysaccharides and proteinaceous components as detected by nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FTIR) spectroscopy. More conventional chemical assays on the biofilm ECM confirmed the presence of these components and also DNA. Proteomics revealed eighteen proteins present in biofilm ECM that were not detected in planktonic bacteria. One ECM protein was unique to 24-h biofilms, two were found only in 96-h biofilms, and fifteen were present in the ECM of both 24- and 96-h NTHi biofilms. All proteins identified were either associated with bacterial membranes or cytoplasmic proteins. Immunocytochemistry showed two of the identified proteins, a DNA-directed RNA polymerase and the outer membrane protein OMP P2, associated with bacteria and biofilm ECM. Identification of biofilm-specific proteins present in immature biofilms is an important step in understanding the in vitro process of NTHi biofilm formation. The presence of a cytoplasmic protein and a membrane protein in the biofilm ECM of immature NTHi biofilms suggests that bacterial cell lysis may be a feature of early biofilm formation.

  16. Anodic Stripping Voltammetry: An Instrumental Analysis Experiment.

    Science.gov (United States)

    Wang, Joseph

    1983-01-01

    Describes an experiment designed to acquaint students with the theory and applications of anodic stripping voltammetry (ASV) as well as such ASV problems as contamination associated with trace analysis. The experimental procedure, instrumentation, and materials discussed are designed to minimize cost and keep procedures as simple as possible. (JM)

  17. Linear Sweep Voltammetry of Adsorbed Neutral Red.

    Science.gov (United States)

    1982-05-01

    E. Creager, G. T. Marks, D. A. Aikens and H. H. Richtol Prepared for Publication in Journal of Electroanalytical Chemistry Rensselaer Polytechnic... Electroanalytical Chemistry It. KEY WORDS (Continue oun reverse side It necessary mid Ideneliy by block ntaibor) Neutral Red, cyclic voltammetry, adsorbed dye 20

  18. Steady state oxygen reduction and cyclic voltammetry

    DEFF Research Database (Denmark)

    Rossmeisl, Jan; Karlberg, Gustav; Jaramillo, Thomas;

    2008-01-01

    The catalytic activity of Pt and Pt3Ni for the oxygen reduction reaction is investigated by applying a Sabatier model based on density functional calculations. We investigate the role of adsorbed OH on the activity, by comparing cyclic voltammetry obtained from theory with previously published...

  19. Voltammetry at porous electrodes: A theoretical study

    CERN Document Server

    Barnes, Edward O; Li, Peilin; Compton, Richard G

    2014-01-01

    Theory is presented to simulate both chronoamperometry and cyclic voltammetry at porous electrodes fabricated by means of electro-deposition around spherical templates. A theoretical method to extract heterogeneous rate constants for quasireversible and irreversible systems is proposed by the approximation of decoupling of the diffusion within the porous electrode and of bulk diffusion to the electrode surface.

  20. Differential pulse voltammetry and additive differential pulse voltammetry with solvent polymeric membrane ion sensors.

    Science.gov (United States)

    Ortuño, J A; Serna, C; Molina, A; Gil, A

    2006-12-01

    The ion transfer across the water-solvent polymeric membrane interface is investigated by using a new device based on a modification of a commercial ion-selective electrode body that permits the accommodation of a platinum counter electrode inside the inner filling solution compartment and, therefore, use of a four-electrode potentiostat with ohmic drop compensation. This device is used here to apply two different double potential pulse techniques--differential pulse voltammetry and additive differential pulse voltammetry--which are more advantageous than other voltammetric techniques, such as normal pulse voltammetry or cyclic voltammetry, for the determination of the characteristic electrochemical parameters of the system. This is due to the concurrence of two factors in these double potential pulse techniques, the peak-shaped response together with a considerable reduction of undesirable current contributions.

  1. Cyclic Voltammetry And Linear Sweep Voltammetry Study Of Cyclic Tertiary Amines

    Institute of Scientific and Technical Information of China (English)

    ZHAO Ping; TIAN JinPing; YIN YingWu

    2001-01-01

    @@ Cyclic six membered a-aminonitrile have proved to be very versatile synthetic intermediates and have been widely used in the construction of a large number of indole alkaloids. In order to obtain some information about the mechanisn of electrochemical synthesis of aaminonitrile. Electrochemistry behaviors that include cyclic voltammetry and linear sweep voltammetry of cyclic tertiary amines which including N-benzylpiperidine (NBP), 1-(l-Methoxycarbonyl ethyl) piperidine (MCEP), N-methylcarbonylppiperidine (NMCP), Nethylpiperidine(NEP) was studied.

  2. Cyclic Voltammetry And Linear Sweep Voltammetry Study Of Cyclic Tertiary Amines

    Institute of Scientific and Technical Information of China (English)

    ZHAO; Ping

    2001-01-01

    Cyclic six membered a-aminonitrile have proved to be very versatile synthetic intermediates and have been widely used in the construction of a large number of indole alkaloids. In order to obtain some information about the mechanisn of electrochemical synthesis of aaminonitrile. Electrochemistry behaviors that include cyclic voltammetry and linear sweep voltammetry of cyclic tertiary amines which including N-benzylpiperidine (NBP), 1-(l-Methoxycarbonyl ethyl) piperidine (MCEP), N-methylcarbonylppiperidine (NMCP), Nethylpiperidine(NEP) was studied.……

  3. Differential linear scan voltammetry: analytical performance in comparison with pulsed voltammetry techniques.

    Science.gov (United States)

    Sheth, Disha B; Gratzl, Miklós

    2013-06-01

    We report here on differential linear scan voltammetry, DLSV, that combines the working principles of linear scan voltammetry, LSV, and the numerous existing pulsed voltammetry techniques. DLSV preserves the information from continuous interrogation in voltage and high accuracy that LSV provides about electrochemical processes, and the much better sensitivity of differential pulsed techniques. DLSV also minimizes the background current compared to both LSV and pulsed voltammetry. An early version of DLSV, derivative stationary electrode polarography, DSEP, had been proposed in the 1960s but soon abandoned in favor of the emerging differential pulsed techniques. Relative to DSEP, DLSV takes advantage of the flexibility of discrete smoothing differentiation that was not available to early investigators. Also, DSEP had been explored in pure solutions and with reversible electrochemical reactions. DLSV is tested in this work in more challenging experimental contexts: the measurement of oxygen with a carbon fiber microelectrode in buffer, and with a gold microdisc electrode exposed to a live biological preparation. This work compares the analytical performance of DLSV and square wave voltammetry, the most popular pulsed voltammetry technique.

  4. Voltammetry as a Model for Teaching Chemical Instrumentation.

    Science.gov (United States)

    Gunasingham, H.; Ang, K. P.

    1985-01-01

    Voltammetry is used as a model for teaching chemical instrumentation to chemistry undergraduates at the National University of Singapore. Lists six criteria used to select a successful teaching model and shows how voltammetry satisfies each criterion. (JN)

  5. Growing and Analyzing Biofilms in Flow Chambers

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim; Sternberg, Claus

    2011-01-01

    This unit describes the setup of flow chamber systems for the study of microbial biofilms, and methods for the analysis of structural biofilm formation. Use of flow chambers allows direct microscopic investigation of biofilm formation. The biofilms in flow chambers develop under hydrodynamic...... conditions, and the environment can be carefully controlled and easily changed. The protocols in this unit include construction of the flow chamber and the bubble trap, assembly and sterilization of the flow chamber system, inoculation of the flow chambers, running of the system, image capture and analysis...

  6. Growing and analyzing biofilms in flow chambers

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim; Sternberg, Claus

    2011-01-01

    This unit describes the setup of flow chamber systems for the study of microbial biofilms, and methods for the analysis of structural biofilm formation. Use of flow chambers allows direct microscopic investigation of biofilm formation. The biofilms in flow chambers develop under hydrodynamic...... conditions, and the environment can be carefully controlled and easily changed. The protocols in this unit include construction of the flow chamber and the bubble trap, assembly and sterilization of the flow chamber system, inoculation of the flow chambers, running of the system, image capture and analysis...

  7. Voltammetry at the Thin-Film Mercury Electrode (TFME).

    Science.gov (United States)

    Pomeroy, R. S.; And Others

    1989-01-01

    Reviewed is the use of the Thin-Film Mercury Electrode for anodic stripping voltammetry, simple voltammetry of solution cations and cathodic stripping voltammetry for the determination of an environmentally important molecule, thiourea. The construction of a simple potentiostat and applications for student laboratory courses are included. (CW)

  8. Reliability of Haemophilus influenzae biofilm measurement via static method, and determinants of in vitro biofilm production.

    Science.gov (United States)

    Obaid, Najla A; Tristram, Stephen; Narkowicz, Christian K; Jacobson, Glenn A

    2016-12-01

    Information is lacking regarding the precision of microtitre plate (MTP) assays used to measure biofilm. This study investigated the precision of an MTP assay to measure biofilm production by nontypeable Haemophilus influenzae (NTHi) and the effects of frozen storage and inoculation technique on biofilm production. The density of bacterial final growth was determined by absorbance after 18-20 h incubation, and biofilm production was then measured by absorbance after crystal violet staining. Biofilm formation was categorised as high and low for each strain. For the high biofilm producing strains of NTHi, interday reproducibility of NTHi biofilm formation measured using the MTP assay was excellent and met the acceptance criteria, but higher variability was observed in low biofilm producers. Method of inoculum preparation was a determinant of biofilm formation with inoculum prepared directly from solid media showing increased biofilm production for at least one of the high producing strains. In general, storage of NTHi cultures at -80 °C for up to 48 weeks did not have any major effect on their ability to produce biofilm.

  9. Evaluation of homogeneous electrocatalysts by cyclic voltammetry.

    Science.gov (United States)

    Rountree, Eric S; McCarthy, Brian D; Eisenhart, Thomas T; Dempsey, Jillian L

    2014-10-06

    The pursuit of solar fuels has motivated extensive research on molecular electrocatalysts capable of evolving hydrogen from protic solutions, reducing CO2, and oxidizing water. Determining accurate figures of merit for these catalysts requires the careful and appropriate application of electroanalytical techniques. This Viewpoint first briefly presents the fundamentals of cyclic voltammetry and highlights practical experimental considerations before focusing on the application of cyclic voltammetry for the characterization of electrocatalysts. Key metrics for comparing catalysts, including the overpotential (η), potential for catalysis (E(cat)), observed rate constant (k(obs)), and potential-dependent turnover frequency, are discussed. The cyclic voltammetric responses for a general electrocatalytic one-electron reduction of a substrate are presented along with methods to extract figures of merit from these data. The extension of this analysis to more complex electrocatalytic schemes, such as those responsible for H2 evolution and CO2 reduction, is then discussed.

  10. Applications of convolution voltammetry in electroanalytical chemistry.

    Science.gov (United States)

    Bentley, Cameron L; Bond, Alan M; Hollenkamp, Anthony F; Mahon, Peter J; Zhang, Jie

    2014-02-18

    The robustness of convolution voltammetry for determining accurate values of the diffusivity (D), bulk concentration (C(b)), and stoichiometric number of electrons (n) has been demonstrated by applying the technique to a series of electrode reactions in molecular solvents and room temperature ionic liquids (RTILs). In acetonitrile, the relatively minor contribution of nonfaradaic current facilitates analysis with macrodisk electrodes, thus moderate scan rates can be used without the need to perform background subtraction to quantify the diffusivity of iodide [D = 1.75 (±0.02) × 10(-5) cm(2) s(-1)] in this solvent. In the RTIL 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, background subtraction is necessary at a macrodisk electrode but can be avoided at a microdisk electrode, thereby simplifying the analytical procedure and allowing the diffusivity of iodide [D = 2.70 (±0.03) × 10(-7) cm(2) s(-1)] to be quantified. Use of a convolutive procedure which simultaneously allows D and nC(b) values to be determined is also demonstrated. Three conditions under which a technique of this kind may be applied are explored and are related to electroactive species which display slow dissolution kinetics, undergo a single multielectron transfer step, or contain multiple noninteracting redox centers using ferrocene in an RTIL, 1,4-dinitro-2,3,5,6-tetramethylbenzene, and an alkynylruthenium trimer, respectively, as examples. The results highlight the advantages of convolution voltammetry over steady-state techniques such as rotating disk electrode voltammetry and microdisk electrode voltammetry, as it is not restricted by the mode of diffusion (planar or radial), hence removing limitations on solvent viscosity, electrode geometry, and voltammetric scan rate.

  11. Voltammetry: mathematical modelling and Inverse Problem

    CERN Document Server

    Koshev, N A; Kuzina, V V

    2016-01-01

    We propose the fast semi-analytical method of modelling the polarization curves in the voltammetric experiment. The method is based on usage of the special func- tions and shows a big calculation speed and a high accuracy and stability. Low computational needs of the proposed algorithm allow us to state the set of Inverse Problems of voltammetry for the reconstruction of metal ions concentrations or the other parameters of the electrolyte under investigation.

  12. Fusion protein-based biofilm fabrication composed of recombinant azurin–myoglobin for dual-level biomemory application

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Taek [Research Institute for Basic Science, Sogang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Chung, Yong-Ho; Yoon, Jinho [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of); Min, Junhong [School of Integrative Engineering, Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Choi, Jeong-Woo, E-mail: jwchoi@sogang.ac.kr [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of)

    2014-11-30

    Graphical abstract: - Highlights: • We developed the fusion protein-based biofilm on the inorganic surface. • For making the fusion protein, the recombinant azurin and the myoglobin was conjugated by the native chemical ligation method. • The developed fusion protein shows unique electrochemical property. • The proposed fusion protein biofilm appears to be a good method for dual-level biomemory device. - Abstract: In the present study, a fusion protein-based biofilm composed of a recombinant azurin–myoglobin (Azu-Myo) has been developed and confirmed its original electrochemical property for dual-level biomemory device application. For this purpose, the azurin was modified with cysteine residues for direct immobilization and conjugation. Then, the recombinant azurin was conjugated with the myoglobin via a sulfo-SMCC bifunctional linker using the chemical ligation method (CLM). The SDS-PAGE and UV–vis spectroscopy were performed to examine the fusion protein conjugates. The prepared Azu-Myo fusion protein was self-assembled onto Au substrate for the biofilm fabrication. Then, the atomic force microscopy (AFM) was used to confirm the immobilization and the surface-enhanced Raman spectroscopy (SERS) was carried out to the surface analysis. Also, the cyclic voltammetry (CV) was carried out to observe an electrochemical property of fabricated biofilm. As a result, the two pair of redox potential values was obtained for dual-level biomemory device application. Then, the dual-level biomemory function was verified by the multi-potential chronoamperometry (MPCA). The results indicate a new fabrication method and material combination for advances in bioelectronic device development.

  13. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci.

    Science.gov (United States)

    Stepanović, Srdjan; Vuković, Dragana; Hola, Veronika; Di Bonaventura, Giovanni; Djukić, Slobodanka; Cirković, Ivana; Ruzicka, Filip

    2007-08-01

    The details of all steps involved in the quantification of biofilm formation in microtiter plates are described. The presented protocol incorporates information on assessment of biofilm production by staphylococci, gained both by direct experience as well as by analysis of methods for assaying biofilm production. The obtained results should simplify quantification of biofilm formation in microtiter plates, and make it more reliable and comparable among different laboratories.

  14. Antimicrobial susceptibility testing in biofilm-growing bacteria.

    Science.gov (United States)

    Macià, M D; Rojo-Molinero, E; Oliver, A

    2014-10-01

    Biofilms are organized bacterial communities embedded in an extracellular polymeric matrix attached to living or abiotic surfaces. The development of biofilms is currently recognized as one of the most relevant drivers of persistent infections. Among them, chronic respiratory infection by Pseudomonas aeruginosa in cystic fibrosis patients is probably the most intensively studied. The lack of correlation between conventional susceptibility test results and therapeutic success in chronic infections is probably a consequence of the use of planktonically growing instead of biofilm-growing bacteria. Therefore, several in vitro models to evaluate antimicrobial activity on biofilms have been implemented over the last decade. Microtitre plate-based assays, the Calgary device, substratum suspending reactors and the flow cell system are some of the most used in vitro biofilm models for susceptibility studies. Likewise, new pharmacodynamic parameters, including minimal biofilm inhibitory concentration, minimal biofilm-eradication concentration, biofilm bactericidal concentration, and biofilm-prevention concentration, have been defined in recent years to quantify antibiotic activity in biofilms. Using these parameters, several studies have shown very significant quantitative and qualitative differences for the effects of most antibiotics when acting on planktonic or biofilm bacteria. Nevertheless, standardization of the procedures, parameters and breakpoints, by official agencies, is needed before they are implemented in clinical microbiology laboratories for routine susceptibility testing. Research efforts should also be directed to obtaining a deeper understanding of biofilm resistance mechanisms, the evaluation of optimal pharmacokinetic/pharmacodynamic models for biofilm growth, and correlation with clinical outcome.

  15. The in vivo biofilm

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Alhede, Maria; Alhede, Morten

    2013-01-01

    Bacteria can grow and proliferate either as single, independent cells or organized in aggregates commonly referred to as biofilms. When bacteria succeed in forming a biofilm within the human host, the infection often becomes very resistant to treatment and can develop into a chronic state. Biofilms...... have been studied for decades using various in vitro models, but it remains debatable whether such in vitro biofilms actually resemble in vivo biofilms in chronic infections. In vivo biofilms share several structural characteristics that differ from most in vitro biofilms. Additionally, the in vivo...... experimental time span and presence of host defenses differ from chronic infections and the chemical microenvironment of both in vivo and in vitro biofilms is seldom taken into account. In this review, we discuss why the current in vitro models of biofilms might be limited for describing infectious biofilms...

  16. Electrochemical Studies of 1,4-Bis[2-(2-pyridyl)-vinyl] Benzene and 1,4-Bis[2-(4-pyridyl) vinyl] Benzene Laser Dyes via Cyclic Voltammetry, Convolutive Voltammetry and Digital Simulation Methods

    Institute of Scientific and Technical Information of China (English)

    EL-DALY, Samy A; EL-HALLAG,Ibrahirn S; EBEED, Ezeiny M; GHONEIM, Mohamed M

    2009-01-01

    Electrochemical properties of two diolefinic laser dyes namely 1,4-bis[2-(2-pyddyl)-vinyl] benzene (2PVB) and 1,4-bis[2-(4-pyridyl) vinyl] benzene (4PVB) have been investigated using cyclic voltammetry and convolutive voltammetry combined with digital simulation at a platinum electrode in 0.1 mol/L tetrabutyl ammonium perchlo-rate (TBAP) in the two different solvents acetonitrile (CH3CN) and dimethylformamide (DMF). The species were reduced via consumption of two sequential electrons to form radical anion and dianion. In switching the potential to positive direction, the two compounds were oxidized by loss of one electron, which was followed by a fast isomeri-sation process. The electrode reaction pathway and the electrochemical parameters of the investigated compounds were determined using cyclic voltammetry. The extracted electrochemical parameters were verified and confirmed via digital simulation and convolutive voltammetry methods.

  17. Comparative transcriptional profiling reveals differential expression of pathways directly and indirectly influencing biofilm formation in Escherichia coli O157:H7

    Science.gov (United States)

    Introduction: Escherichia coli O157:H7 (O157) is a frequent cause of foodborne disease outbreaks. O157 encodes virulence factors for colonizing and survival in reservoir animals and the environment. For example, genetic factors promoting biofilm formation are linked to survival of O157 in and outsid...

  18. Mesoscale Elucidation of Biofilm Shear Behavior

    CERN Document Server

    Barai, Pallab; Mukherjee, Partha P

    2015-01-01

    Formation of bacterial colonies as biofilm on the surface/interface of various objects has the potential to impact not only human health and disease but also energy and environmental considerations. Biofilms can be regarded as soft materials, and comprehension of their shear response to external forces is a key element to the fundamental understanding. A mesoscale model has been presented in this article based on digitization of a biofilm microstructure. Its response under externally applied shear load is analyzed. Strain stiffening type behavior is readily observed under high strain loads due to the unfolding of chains within soft polymeric substrate. Sustained shear loading of the biofilm network results in strain localization along the diagonal direction. Rupture of the soft polymeric matrix can potentially reduce the intercellular interaction between the bacterial cells. Evolution of stiffness within the biofilm network under shear reveals two regions: a) initial increase in stiffness due to strain stiffe...

  19. Pseudomonas biofilms: possibilities of their control.

    Science.gov (United States)

    Masák, Jan; Čejková, Alena; Schreiberová, Olga; Rezanka, Tomáš

    2014-07-01

    Genus Pseudomonas includes a large number of species that can be encountered in biotechnological processes as well as in the role of serious human or plant pathogens. Pseudomonads easily form biofilms on various types of surfaces. The biofilm phenotype is characterized by an increased resistance to environmental influences including resistance to antibiotics and other disinfectants, causing a number of problems in health care, food industry, and other areas. Considerable attention is therefore paid to the possibilities of eradication/destruction of pseudomonads biofilms both in terms of understanding the mechanisms of biofilm formation and at the level of finding suitable antibiofilm tools applicable in practice. The first part of this review is devoted to an overview of the regulatory mechanisms that are directly or indirectly involved in the formation of biofilm. The most effective approaches to suppressing the formation of biofilm that do not cause the development of resistance are based on the application of substances that interfere with the regulatory molecules or block the appropriate regulatory mechanisms involved in biofilm development by the cells. Pseudomonads biofilm formation is, similar to other microorganisms, a sophisticated process with many regulatory elements. The suppression of this process therefore also requires multiple antibiofilm tools.

  20. Biofilm Fixed Film Systems

    Directory of Open Access Journals (Sweden)

    Dipesh Das

    2011-09-01

    Full Text Available The work reviewed here was published between 2008 and 2010 and describes research that involved aerobic and anoxic biofilm treatment of water pollutants. Biofilm denitrification systems are covered when appropriate. References catalogued here are divided on the basis of fundamental research area or reactor types. Fundamental research into biofilms is presented in two sections, Biofilm Measurement and Characterization and Growth and Modeling. The reactor types covered are: trickling filters, rotating biological contactors, fluidized bed bioreactors, submerged bed biofilm reactors, biological granular activated carbon, membrane bioreactors, and immobilized cell reactors. Innovative reactors, not easily classified, are then presented, followed by a section on biofilms on sand, soil and sediment.

  1. Corrosive electrochemistry of jamesonite by cyclic voltammetry

    Institute of Scientific and Technical Information of China (English)

    余润兰; 胡岳华; 邱冠周; 覃文庆

    2004-01-01

    The corrosive electrochemistry of jamesonite was studied by cyclic voltammetry. Every peak in voltammograms was identified through thermodynamic calculation. The results show an irreversible electrode process by the strong adsorption of oxidation elemental sulfur on jamesonite. A deficient-metal and sulfur-rich compound is formed under the potential of 80 mV at pH 6.86. The passive action by elemental sulfur occurs from 80 to 470 mV and S2O23- , SO24- are produced at potential over 470 mV. The anodic peak producing SO24- is inhibited due to the deposition of PbSO4 at higher potential in Na2SO4 solution. The corrosive action of jamesonite becomes strong and the redox characterization similar to PbS, FeS and Sb2 S3 appears at pH 9.18.

  2. Asymmetric Marcus-Hush theory for voltammetry.

    Science.gov (United States)

    Laborda, Eduardo; Henstridge, Martin C; Batchelor-McAuley, Christopher; Compton, Richard G

    2013-06-21

    The current state-of-the-art in modeling the rate of electron transfer between an electroactive species and an electrode is reviewed. Experimental studies show that neither the ubiquitous Butler-Volmer model nor the more modern symmetric Marcus-Hush model are able to satisfactorily reproduce the experimental voltammetry for both solution-phase and surface-bound redox couples. These experimental deviations indicate the need for revision of the simplifying approximations used in the above models. Within this context, models encompassing asymmetry are considered which include different vibrational and solvation force constants for the electroactive species. The assumption of non-adiabatic electron transfer is also examined. These refinements have provided more satisfactory models of the electron transfer process and they enable us to gain more information about the microscopic characteristics of the system by means of simple electrochemical measurements.

  3. Prediction of optimal biofilm thickness for membrane-attached biofilms growing in an extractive membrane bioreactor.

    Science.gov (United States)

    Pavasant, P; Dos Santos, L M; Pistikopoulos, E N; Livingston, A G

    1996-11-05

    This article presents a mathematical model of membrane-attached biofilm (MAB) behavior in a single-tube extractive membrane bioreactor (STEMB). MABs can be used for treatment of wastewaters containing VOCs, treatment of saline wastewaters, and nitrification processes. Extractive membrane bioreactors (EMBs) are employed to prevent the direct contact between a toxic volatile pollutant and the aerated gas by allowing counterdiffusion of substrates; i.e., pollutant diffuses from the tube side into the biofilm, whereas oxygen diffuses from the shell side into the biofilm. This reduces the air stripping problems usually found in conventional bioreactors. In this study, the biodegradation of a toxic VOC (1,2-dichloroethane, DCE) present in a synthetic wastewater has been investigated. An unstructured model is used to describe cell growth and cell decay in the MAB. The model has been verified by comparing model predicted trends with experimental data collected over 5 to 20-day periods, and has subsequently been used to model steady states in biofilm behavior over longer time scales. The model is capable of predicting the correct trends in system variables such as biofilm thickness, DCE flux across the membrane, carbon dioxide evolution, and suspended biomass. Steady states (constant biofilm thickness and DCE flux) are predicted, and factors that affect these steady states, i.e., cell endogeneous decay rate, and biofilm attrition, are investigated. Biofilm attrition does not have a great influence on biofilm behavior at low values of detachment coefficient close to those typically reported in the literature. Steady-state biofilm thickness is found to be an important variable; a thin biofilm results in a high DCE flux across the membrane, but with the penalty of a high loss of DCE via air stripping. The optimal biofilm thickness at steady state can be determined by trading off the decrease in air stripping (desirable) and the decrease in DCE flux (undesirable) which occur

  4. A novel assay of biofilm antifungal activity reveals that amphotericin B and caspofungin lyse Candida albicans cells in biofilms.

    Science.gov (United States)

    DiDone, Louis; Oga, Duana; Krysan, Damian J

    2011-08-01

    The ability of Candida albicans to form drug-resistant biofilms is an important factor in its contribution to human disease. Assays to identify and characterize molecules with activity against fungal biofilms are crucial for the development of drugs with improved anti-biofilm activity. Here we report the application of an adenylate kinase (AK)-based cytotoxicity assay of fungal cell lysis to the characterization of agents active against C. albicans biofilms. We have developed three protocols for the AK assay. The first measures AK activity in the supernatants of biofilms treated with antifungal drugs and can be performed in parallel with a standard 2,3-bis-(2-methoxy-4-nitro-5-sulphophenyl)-2H-tetrazolium-5-caboxanilide-based biofilm susceptibility assay; a second, more sensitive protocol measures the AK activity present within the biofilm matrix; and a third procedure allows the direct visualization of lytic activity toward biofilms formed on catheter material. Amphotericin B and caspofungin, the two most effective anti-biofilm drugs currently used to treat fungal infections, both directly lyse planktonic C. albicans cells in vitro, leading to the release of AK into the culture medium. These studies serve to validate the AK-based lysis assay as a useful addition to the methods for the characterization of antifungal agents active toward biofilms and provide insights into the mode of action of amphotericin B and caspofungin against C. albicans biofilms.

  5. Pseudomonas aeruginosa Biofilm Infections

    DEFF Research Database (Denmark)

    Rybtke, Morten; Hultqvist, Louise Dahl; Givskov, Michael

    2015-01-01

    Studies of biopsies from infectious sites, explanted tissue and medical devises have provided evidence that biofilms are the underlying cause of a variety of tissue-associated and implant-associated recalcitrant human infections. With a need for novel anti-biofilm treatment strategies, research...... in biofilm infection microbiology, biofilm formation mechanisms and biofilm-associated antimicrobial tolerance has become an important area in microbiology. Substantial knowledge about biofilm formation mechanisms, biofilm-associated antimicrobial tolerance and immune evasion mechanisms has been obtained...... through work with biofilms grown in in vitro experimental setups, and the relevance of this information in the context of chronic infections is being investigated by the use of animal models of infection. Because our current in vitro experimental setups and animal models have limitations, new advanced...

  6. Biofilms: A microbial home

    Science.gov (United States)

    Chandki, Rita; Banthia, Priyank; Banthia, Ruchi

    2011-01-01

    Microbial biofilms are mainly implicated in etiopathogenesis of caries and periodontal disease. Owing to its properties, these pose great challenges. Continuous and regular disruption of these biofilms is imperative for prevention and management of oral diseases. This essay provides a detailed insight into properties, mechanisms of etiopathogenesis, detection and removal of these microbial biofilms. PMID:21976832

  7. Biofilms: A microbial home

    OpenAIRE

    Chandki, Rita; Banthia, Priyank; Banthia, Ruchi

    2011-01-01

    Microbial biofilms are mainly implicated in etiopathogenesis of caries and periodontal disease. Owing to its properties, these pose great challenges. Continuous and regular disruption of these biofilms is imperative for prevention and management of oral diseases. This essay provides a detailed insight into properties, mechanisms of etiopathogenesis, detection and removal of these microbial biofilms.

  8. The Biofilm Challenge

    DEFF Research Database (Denmark)

    Alhede, Maria; Alhede, Morten

    2014-01-01

    The concept of biofilms has emerged in the clinical setting during the last decade. Infections involving biofilms have been documented in all parts of the human body, and it is currently believed that the presence of biofilm-forming bacteria is equivalent to chronic infection. A quick Pubmed search...

  9. Cyclic voltammetry of fast conducting electrocatalytic films.

    Science.gov (United States)

    Costentin, Cyrille; Savéant, Jean-Michel

    2015-07-15

    In the framework of contemporary energy challenges, cyclic voltammetry is a particularly useful tool for deciphering the kinetics of catalytic films. The case of fast conducting films is analyzed, whether conduction is of the ohmic type or proceeds through rapid electron hopping. The rate-limiting factors are then the diffusion of the substrate in solution and through the film as well as the catalytic reaction itself. The dimensionless combination of the characteristics of these factors allows reducing the number of actual parameters to a maximum of two. The kinetics of the system may then be fully analyzed with the help of a kinetic zone diagram. Observing the variations of the current-potential responses with operational parameters such as film thickness, the potential scan rate and substrate concentration allows a precise assessment of the interplay between these factors and of the values of the rate controlling factors. A series of thought experiments is described in order to render the kinetic analysis more palpable.

  10. Anodic stripping voltammetry enhancement by redox magnetohydrodynamics.

    Science.gov (United States)

    Clark, Emily A; Fritsch, Ingrid

    2004-04-15

    The effect of an external magnetic field on linear scan anodic stripping voltammetry (ASV) in solutions of 10(-6)-10(-7) M concentrations of lead, cadmium, and copper at mercury films on glassy carbon electrodes has been investigated. A high concentration of Hg(2+) was added to the analyte solution to induce a large cathodic current during the deposition step. Therefore, a large Lorentz force from the net flux of charge through the magnetic field resulted in convection due to magnetohydrodynamics. The faster delivery of analytes to the mercury film electrode during deposition caused an increase in the anodic stripping peaks. The effect of varying Hg(2+) concentrations (0-60 mM) and magnetic field strengths (0-1.77 T) on the enhancement of the stripping peaks was investigated. Enhancements as large as 129% for peak currents and 167% for peak areas were observed. An enhancement of approximately 100% was observed when 60 mM Fe(3+) replaced high concentrations of Hg(2+). This method of convection exhibits promise for small-volume ASV analysis with possible improved limits of detection and decreased preconcentration times.

  11. Fast Selective Detection of Pyocyanin Using Cyclic Voltammetry.

    Science.gov (United States)

    Alatraktchi, Fatima AlZahra'a; Andersen, Sandra Breum; Johansen, Helle Krogh; Molin, Søren; Svendsen, Winnie E

    2016-03-19

    Pyocyanin is a virulence factor uniquely produced by the pathogen Pseudomonas aeruginosa. The fast and selective detection of pyocyanin in clinical samples can reveal important information about the presence of this microorganism in patients. Electrochemical sensing of the redox-active pyocyanin is a route to directly quantify pyocyanin in real time and in situ in hospitals and clinics. The selective quantification of pyocyanin is, however, limited by other redox-active compounds existing in human fluids and by other metabolites produced by pathogenic bacteria. Here we present a direct selective method to detect pyocyanin in a complex electroactive environment using commercially available electrodes. It is shown that cyclic voltammetry measurements between -1.0 V to 1.0 V reveal a potential detection window of pyocyanin of 0.58-0.82 V that is unaffected by other redox-active interferents. The linear quantification of pyocyanin has an R² value of 0.991 across the clinically relevant concentration range of 2-100 µM. The proposed method was tested on human saliva showing a standard deviation of 2.5% ± 1% (n = 5) from the known added pyocyanin concentration to the samples. This inexpensive procedure is suggested for clinical use in monitoring the presence and state of P. aeruginosa infection in patients.

  12. Cyclic voltammetry of apple fruits: Memristors in vivo.

    Science.gov (United States)

    Volkov, Alexander G; Nyasani, Eunice K; Tuckett, Clayton; Blockmon, Avery L; Reedus, Jada; Volkova, Maya I

    2016-12-01

    A memristor is a resistor with memory that exhibits a pinched hysteretic relationship in cyclic voltammetry. Recently, we have found memristors in the electrical circuitry of plants and seeds. There are no publications in literature about the possible existence of memristors and electrical differentiators in fruits. Here we found that the electrostimulation of Golden Delicious or Arkansas Black apple fruits by bipolar periodic waves induces hysteresis loops with pinched points in cyclic voltammograms at low frequencies between 0.1MHz and 1MHz. At high frequencies of 1kHz, the pinched hysteresis loop transforms to a non-pinched hysteresis loop instead of a single line I=V/R for ideal memristors because the amplitude of electrical current depends on capacitance of a fruit's tissue and electrodes, frequency and direction of scanning. Electrostimulation of electrical circuits in apple fruits by periodic voltage waves also induces electrotonic potential propagation due to cell-to-cell electrical coupling with electrical differentiators. A differentiator is an electrical circuit in which the output of the circuit is approximately directly proportional to the rate of change of the input. The information gained from electrostimulation can be used to elucidate and to observe electrochemical and electrophysiological properties of electrical circuits in fruits.

  13. Voltammetry of redox analytes at trace concentrations with nanoelectrode ensembles.

    Science.gov (United States)

    Moretto, Ligia Maria; Pepe, Niki; Ugo, Paolo

    2004-04-19

    Gold nanoelectrodes ensembles (NEEs) have been prepared by electroless plating of Au nanoelectrode elements within the pores of a microporous polycarbonate template membrane. Cyclic voltammograms recorded in (ferrocenylmethyl) trimethylammonium hexafluorophosphate (FA(+) PF(6)(-)) solutions showed that these NEEs operate in the "total-overlap" response regime, giving well resolved peak shaped voltammograms. Experimental results show that the faradaic/background currents ratios at the NEE are independent on the total geometric area of the ensemble, so that NEE can be enlarged or miniaturized at pleasure without influencing the very favorable signal/noise ratio. Differential pulse voltammetry (DPV) at the NEE is optimized for direct determinations at trace levels. DPV at NEE allowed the determination (with no preconcentration) of trace amounts of FA(+), with a detection limit of 0.02muM. The use of NEE and DPV in cytochrome c (cyt c) solutions showed the possibility to observe the direct electrochemistry of submicromolar concentration of the protein, even without the need of adding any promoter or mediator.

  14. Development and calibration of a microfluidic biofilm growth cell with flow-templating and multi-modal characterization.

    Science.gov (United States)

    Paquet-Mercier, Francois; Karas, Adnane; Safdar, Muhammad; Aznaveh, Nahid Babaei; Zarabadi, Mirpouyan; Greener, Jesse

    2014-01-01

    We report the development of a microfluidic flow-templating platform with multi-modal characterization for studies of biofilms and their precursor materials. A key feature is a special three inlet flow-template compartment, which confines and controls the location of biofilm growth against a template wall. Characterization compartments include Raman imaging to study the localization of the nutrient solutions, optical microscopy to quantify biofilm biomass and localization, and cyclic voltammetry for flow velocity measurements. Each compartment is tested and then utilized to make preliminary measurements.

  15. Porosity Study of Hybrid Silica Mesostructure in Aluminium Oxide Membrane Columnar by Cyclic Voltammetry Method

    Directory of Open Access Journals (Sweden)

    M.N. Jalil

    2011-12-01

    Full Text Available Silica mesostructure has been grown within with a porous aluminium oxide membrane columnar material (hybrid-AOM. This was prepared using a sol-gel technique with Pluronic P123 triblock copolymer as the structure-directing agent and tetraethyl orthosilicate as the inorganic source. The porosity of the hybrid-AOM after ethanol extraction was calculated from the cyclic voltammetry response of a neutral probe (FcMeOH, using Randles-Sevčik equation.

  16. Stoichiometry and Formation Constant Determination by Linear Sweep Voltammetry.

    Science.gov (United States)

    Schultz, Franklin A.

    1979-01-01

    In this paper an experiment is described in which the equilibrium constants necessary for determining the composition and distribution of lead (II)-oxalate species may be measured by linear sweep voltammetry. (Author/BB)

  17. Optimizing future treatment of enterococcal infections: attacking the biofilm?

    Science.gov (United States)

    Paganelli, Fernanda L; Willems, Rob J; Leavis, Helen L

    2012-01-01

    Enterococcus faecalis and Enterococcus faecium are among the leading causative agents of nosocomial infections and are infamous for their resistance to many antibiotics. They cause difficult-to-treat infections, often originating from biofilm-mediated infections associated with implanted medical devices or endocarditis. Biofilms protect bacteria against antibiotics and phagocytosis, and physical removal of devices or infected tissue is often needed but is frequently not possible. Currently there are no clinically available compounds that disassemble biofilms. In this review we discuss all known structural and regulatory genes involved in enterococcal biofilm formation, the compounds directed against biofilm formation that have been studied, and potentially useful targets for future drugs to treat enterococcal biofilm-associated infections.

  18. An electrochemical impedance model for integrated bacterial biofilms

    Energy Technology Data Exchange (ETDEWEB)

    Ben-Yoav, Hadar, E-mail: benyoav@post.tau.ac.il [Department of Physical Electronics, School of Electrical Engineering, Faculty of Engineering, Tel Aviv University (Israel); Freeman, Amihay [Department of Molecular Microbiology and Biotechnology, Faculty of Life Sciences, Tel Aviv University (Israel); Sternheim, Marek [The Center for Nanoscience and Nanotechnology, Tel Aviv University (Israel); Shacham-Diamand, Yosi [Department of Physical Electronics, School of Electrical Engineering, Faculty of Engineering, Tel Aviv University (Israel)

    2011-09-30

    Bacterial cells attachment onto solid surfaces and the following growth into mature microbial biofilms may result in highly antibiotic resistant biofilms. Such biofilms may be incidentally formed on tissues or implanted devices, or intentionally formed by directed deposition of microbial sensors on whole-cell bio-chip surface. A new method for electrical characterization of the later on-chip microbial biofilm buildup is presented in this paper. Measurement of impedance vs. frequency in the range of 100 mHz to 400 kHz of Escherichia coli cells attachment to indium-tin-oxide-coated electrodes was carried out while using optical microscopy estimating the electrode area coverage. We show that impedance spectroscopy measurements can be interpreted by a simple electrical equivalent model characterizing both attachment and growth of the biofilm. The correlation of extracted equivalent electrical lumped components with the visual biofilm parameters and their dependence on the attachment and growth phases is confirmed.

  19. Integrated circuit-based electrochemical sensor for spatially resolved detection of redox-active metabolites in biofilms.

    Science.gov (United States)

    Bellin, Daniel L; Sakhtah, Hassan; Rosenstein, Jacob K; Levine, Peter M; Thimot, Jordan; Emmett, Kevin; Dietrich, Lars E P; Shepard, Kenneth L

    2014-01-01

    Despite advances in monitoring spatiotemporal expression patterns of genes and proteins with fluorescent probes, direct detection of metabolites and small molecules remains challenging. A technique for spatially resolved detection of small molecules would benefit the study of redox-active metabolites that are produced by microbial biofilms and can affect their development. Here we present an integrated circuit-based electrochemical sensing platform featuring an array of working electrodes and parallel potentiostat channels. 'Images' over a 3.25 × 0.9 mm(2) area can be captured with a diffusion-limited spatial resolution of 750 μm. We demonstrate that square wave voltammetry can be used to detect, identify and quantify (for concentrations as low as 2.6 μM) four distinct redox-active metabolites called phenazines. We characterize phenazine production in both wild-type and mutant Pseudomonas aeruginosa PA14 colony biofilms, and find correlations with fluorescent reporter imaging of phenazine biosynthetic gene expression.

  20. Integrated circuit-based electrochemical sensor for spatially resolved detection of redox-active metabolites in biofilms

    Science.gov (United States)

    Bellin, Daniel L.; Sakhtah, Hassan; Rosenstein, Jacob K.; Levine, Peter M.; Thimot, Jordan; Emmett, Kevin; Dietrich, Lars E. P.; Shepard, Kenneth L.

    2014-01-01

    Despite advances in monitoring spatiotemporal expression patterns of genes and proteins with fluorescent probes, direct detection of metabolites and small molecules remains challenging. A technique for spatially resolved detection of small molecules would benefit the study of redox-active metabolites produced by microbial biofilms, which can drastically affect colony development. Here we present an integrated circuit-based electrochemical sensing platform featuring an array of working electrodes and parallel potentiostat channels. “Images” over a 3.25 × 0.9 mm area can be captured with a diffusion-limited spatial resolution of 750 μm. We demonstrate that square wave voltammetry can be used to detect, identify, and quantify (for concentrations as low as 2.6 μM) four distinct redox-active metabolites called phenazines. We characterize phenazine production in both wild-type and mutant Pseudomonas aeruginosa PA14 colony biofilms, and find correlations with fluorescent reporter imaging of phenazine biosynthetic gene expression. PMID:24510163

  1. Phenol-degrading anode biofilm with high coulombic efficiency in graphite electrodes microbial fuel cell.

    Science.gov (United States)

    Zhang, Dongdong; Li, Zhiling; Zhang, Chunfang; Zhou, Xue; Xiao, Zhixing; Awata, Takanori; Katayama, Arata

    2017-03-01

    A microbial fuel cell (MFC), with graphite electrodes as both the anode and cathode, was operated with a soil-free anaerobic consortium for phenol degradation. This phenol-degrading MFC showed high efficiency with a current density of 120 mA/m(2) and a coulombic efficiency of 22.7%, despite the lack of a platinum catalyst cathode and inoculation of sediment/soil. Removal of planktonic bacteria by renewing the anaerobic medium did not decrease the performance, suggesting that the phenol-degrading MFC was not maintained by the planktonic bacteria but by the microorganisms in the anode biofilm. Cyclic voltammetry analysis of the anode biofilm showed distinct oxidation and reduction peaks. Analysis of the microbial community structure of the anode biofilm and the planktonic bacteria based on 16S rRNA gene sequences suggested that Geobacter sp. was the phenol degrader in the anode biofilm and was responsible for current generation.

  2. A quantitative method to measure biofilm removal efficiency from complex biomaterial surfaces using SEM and image analysis

    Science.gov (United States)

    Vyas, N.; Sammons, R. L.; Addison, O.; Dehghani, H.; Walmsley, A. D.

    2016-09-01

    Biofilm accumulation on biomaterial surfaces is a major health concern and significant research efforts are directed towards producing biofilm resistant surfaces and developing biofilm removal techniques. To accurately evaluate biofilm growth and disruption on surfaces, accurate methods which give quantitative information on biofilm area are needed, as current methods are indirect and inaccurate. We demonstrate the use of machine learning algorithms to segment biofilm from scanning electron microscopy images. A case study showing disruption of biofilm from rough dental implant surfaces using cavitation bubbles from an ultrasonic scaler is used to validate the imaging and analysis protocol developed. Streptococcus mutans biofilm was disrupted from sandblasted, acid etched (SLA) Ti discs and polished Ti discs. Significant biofilm removal occurred due to cavitation from ultrasonic scaling (p dental implants. This imaging and analysis method will be of value to other researchers and manufacturers wishing to study biofilm growth and removal.

  3. 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...

  4. Degradation in the Dentin-Composite Interface Subjected to Multi-Species Biofilm Challenges

    OpenAIRE

    Li, Yuping; Carrera, Carola; Chen, Ruoqiong; Li, Jianying; Patricia, Lenton; Rudney, Joel D.; Jones, Robert S.; Aparicio, Conrado; Fok, Alex

    2013-01-01

    Oral biofilms can degrade the components in dental resin-based composite restorations, thus compromising marginal integrity and leading to secondary caries. In this study, we investigated the mechanical integrity of the dentin-composite interface challenged with multi-species oral biofilms. While most studies used single-species biofilms, we used a more realistic, diverse biofilm model produced directly from plaques collected from donors with a history of early childhood caries. Dentin–compos...

  5. AFM Structural Characterization of Drinking Water Biofilm under Physiological Conditions

    Science.gov (United States)

    Due to the complexity of mixed culture drinking water biofilm, direct visual observation under in situ conditions has been challenging. In this study, atomic force microscopy (AFM) revealed the three dimensional morphology and arrangement of drinking water relevant biofilm in air...

  6. Studying bacterial multispecies biofilms

    DEFF Research Database (Denmark)

    Røder, Henriette Lyng; Sørensen, Søren Johannes; Burmølle, Mette

    2016-01-01

    and drawbacks of varying the degree of complexity. This review aims to facilitate multispecies biofilm research in order to expand the current limited knowledge on interspecies interactions. Recent technological advances have enabled total diversity analysis of highly complex and diverse microbial communities...... at the microscale of complex communities, including biofilms.Studies of multispecies biofilms and the interactions shaping these are conducted in traditional approaches used for single-species biofilms with some adjustments; but a crucial point for consideration is which strains to combine and where these should...

  7. Study on the Curcumin dynamics and distribution through living biofilms

    Science.gov (United States)

    Carvalho, Mariana T.; Dovigo, Lívia N.; Rastelli, Alessandra N. S.; Bagnato, Vanderlei S.

    2013-03-01

    Human oral cavity is colonized by a wide range of microorganisms, often organized in biofilms. These biofilms are responsible for the pathogenesis of caries and most periodontal diseases. A possible alternative to reduce biofilms is the photodynamic inactivation (PDI). The success of the PDI depends on different factors. The time required by the PS to remain in contact with the target cells prior to illumination is determinant for the technique's efficacy. This study aimed to assess the interaction between the PS and the biofilm prior to the PDI. We used confocal microscopy and FLIM to evaluate the interaction between the PS and the biofilm's microorganism during the pre-irradiation time (PIT). The study of this dynamics can lead to the understanding of why only some PSs are effective and why is necessary a long PIT for some microorganisms. Our results showed that are differences for each PIT. These differences can be the determinate for the efficacy of the PDI. We observed that the microorganism needs time to concentrate and/or transport the PS within the biofilm. We presented preliminary results for biofilms of Candida albicans and Streptococcus mutans in the presence of Curcumin and compared it with the literature. We observed that the effectiveness of the PDI might be directly correlated to the position of the PS with the biofilm. Further analyses will be conducted in order to confirm the potential of FLIM to assess the PS dynamics within the biofilms.

  8. Biofilm detachment mechanisms in a liquid-fluidized bed.

    Science.gov (United States)

    Chang, H T; Rittmann, B E; Amar, D; Heim, R; Ehlinger, O; Lesty, Y

    1991-08-20

    Bed fluidization offers the possibility of gaining the advantages of fixed-film biological processes without the disadvantage of pore clogging. However, the biofilm detachment rate, due to hydrodynamics and particle-to-particle attrition, is very poorly understood for fluidized-bed biofilm processes. In this work, a two-phase fluidized-bed biofilm was operated under a constant surface loading (0.09 mg total organic carbon/cm(2) day) and with a range of bed height (H), fluid velocities (U), and support-particle concentrations (C(p)). Direct measurements were made for the specific biofilm loss rate coefficient (b(s))and the total biofilm accumulation (X(f)L(f)). A hydrodynamic model allowed independent determination of the biofilm density (X(f)), biofilm thickness (L(f)), liquid shear stress (tau), and Reynolds number (Re). Multiple regression analysis of the results showed that increased particle-to-particle attrition, proportional to C(p) and increased turbulence, described by Re, caused the biofilms to be denser and thinner. The specific detachment rate coefficient (b(s)) increased as C(p) and Re increased. Almost all of the 6, values were larger than predicted by a previous model derived for smooth biofilms on a nonfluidized surface. Therefore, the turbulence and attrition of bed fluidization appear to be dominant detachment mechanisms.

  9. Hydraulic resistance of biofilms

    KAUST Repository

    Dreszer, C.

    2013-02-01

    Biofilms may interfere with membrane performance in at least three ways: (i) increase of the transmembrane pressure drop, (ii) increase of feed channel (feed-concentrate) pressure drop, and (iii) increase of transmembrane passage. Given the relevance of biofouling, it is surprising how few data exist about the hydraulic resistance of biofilms that may affect the transmembrane pressure drop and membrane passage. In this study, biofilms were generated in a lab scale cross flow microfiltration system at two fluxes (20 and 100Lm-2h-1) and constant cross flow (0.1ms-1). As a nutrient source, acetate was added (1.0mgL-1 acetate C) besides a control without nutrient supply. A microfiltration (MF) membrane was chosen because the MF membrane resistance is very low compared to the expected biofilm resistance and, thus, biofilm resistance can be determined accurately. Transmembrane pressure drop was monitored. As biofilm parameters, thickness, total cell number, TOC, and extracellular polymeric substances (EPS) were determined, it was demonstrated that no internal membrane fouling occurred and that the fouling layer actually consisted of a grown biofilm and was not a filter cake of accumulated bacterial cells. At 20Lm-2h-1 flux with a nutrient dosage of 1mgL-1 acetate C, the resistance after 4 days reached a value of 6×1012m-1. At 100Lm-2h-1 flux under the same conditions, the resistance was 5×1013m-1. No correlation of biofilm resistance to biofilm thickness was found; Biofilms with similar thickness could have different resistance depending on the applied flux. The cell number in biofilms was between 4×107 and 5×108 cellscm-2. At this number, bacterial cells make up less than a half percent of the overall biofilm volume and therefore did not hamper the water flow through the biofilm significantly. A flux of 100Lm-2h-1 with nutrient supply caused higher cell numbers, more biomass, and higher biofilm resistance than a flux of 20Lm-2h-1. However, the biofilm thickness

  10. Meningococcal biofilm formation

    DEFF Research Database (Denmark)

    Lappann, M.; Haagensen, Janus Anders Juul; Claus, H.

    2006-01-01

    We show that in a standardized in vitro flow system unencapsulated variants of genetically diverse lineages of Neisseria meningitidis formed biofilms, that could be maintained for more than 96 h. Biofilm cells were resistant to penicillin, but not to rifampin or ciprofloxacin. For some strains......, microcolony formation within biofilms was observed. Microcolony formation in strain MC58 depended on a functional copy of the pilE gene encoding the pilus subunit pilin, and was associated with twitching of cells. Nevertheless, unpiliated pilE mutants formed biofilms showing that attachment and accumulation......X alleles was identified among genetically diverse meningococcal strains. PilX alleles differed in their propensity to support autoaggregation of cells in suspension, but not in their ability to support microcolony formation within biofilms in the continuous flow system....

  11. Analytical possibilities of microelectrode use for stripping voltammetry

    Energy Technology Data Exchange (ETDEWEB)

    Matysik, F.M. (Leipzig Univ. (Germany). Dept. of Chemistry); Glaeser, P. (Leipzig Univ. (Germany). Dept. of Chemistry); Werner, G. (Leipzig Univ. (Germany). Dept. of Chemistry)

    1994-07-01

    The analytical utility of microelectrodes for stripping voltammetry is discussed from several points of view. The application of microelectrodes for microanalysis is demonstrated using a novel capillary flow injection system. Heavy metals at [mu]g l[sup -1] concentrations have been determined in [mu]l-samples. The influence of electrode size and convection during the deposition period of anodic stripping voltammetry on the reproducibility of trace metal determination was studied for various types of electrodes. In the case of mercury film microelectrodes, the precision can be improved if the accumulation of the analyte is performed under quiescent conditions. Practical examples of stripping voltammetry with microelectrodes such as copper determination in whisky and trace metal measurements in drinking water are given. (orig.)

  12. Hitchhiker's Guide to Voltammetry: Acute and Chronic Electrodes for in Vivo Fast-Scan Cyclic Voltammetry.

    Science.gov (United States)

    Rodeberg, Nathan T; Sandberg, Stefan G; Johnson, Justin A; Phillips, Paul E M; Wightman, R Mark

    2017-02-15

    Fast-scan cyclic voltammetry (FSCV) has been used for over 20 years to study rapid neurotransmission in awake and behaving animals. These experiments were first carried out with carbon-fiber microelectrodes (CFMs) encased in borosilicate glass, which can be inserted into the brain through micromanipulators and guide cannulas. More recently, chronically implantable CFMs constructed with small diameter fused-silica have been introduced. These electrodes can be affixed in the brain with minimal tissue response, which permits longitudinal measurements of neurotransmission in single recording locations during behavior. Both electrode designs have been used to make novel discoveries in the fields of neurobiology, behavioral neuroscience, and psychopharmacology. The purpose of this Review is to address important considerations for the use of FSCV to study neurotransmitters in awake and behaving animals, with a focus on measurements of striatal dopamine. Common issues concerning experimental design, data collection, and calibration are addressed. When necessary, differences between the two methodologies (acute vs chronic recordings) are discussed. The topics raised in this Review are particularly important as the field moves beyond dopamine toward new neurochemicals and brain regions.

  13. Biofilm Cohesive Strength as a Basis for Biofilm Recalcitrance: Are Bacterial Biofilms Overdesigned?

    Science.gov (United States)

    Aggarwal, Srijan; Stewart, Philip S; Hozalski, Raymond M

    2015-01-01

    Bacterial biofilms are highly resistant to common antibacterial treatments, and several physiological explanations have been offered to explain the recalcitrant nature of bacterial biofilms. Herein, a biophysical aspect of biofilm recalcitrance is being reported on. While engineering structures are often overdesigned with a factor of safety (FOS) usually under 10, experimental measurements of biofilm cohesive strength suggest that the FOS is on the order of thousands. In other words, bacterial biofilms appear to be designed to withstand extreme forces rather than typical or average loads. In scenarios requiring the removal or control of unwanted biofilms, this emphasizes the importance of considering strategies for structurally weakening the biofilms in conjunction with bacterial inactivation.

  14. Involvement of NADH Oxidase in Biofilm Formation in Streptococcus sanguinis.

    Directory of Open Access Journals (Sweden)

    Xiuchun Ge

    Full Text Available Biofilms play important roles in microbial communities and are related to infectious diseases. Here, we report direct evidence that a bacterial nox gene encoding NADH oxidase is involved in biofilm formation. A dramatic reduction in biofilm formation was observed in a Streptococcus sanguinis nox mutant under anaerobic conditions without any decrease in growth. The membrane fluidity of the mutant bacterial cells was found to be decreased and the fatty acid composition altered, with increased palmitic acid and decreased stearic acid and vaccenic acid. Extracellular DNA of the mutant was reduced in abundance and bacterial competence was suppressed. Gene expression analysis in the mutant identified two genes with altered expression, gtfP and Idh, which were found to be related to biofilm formation through examination of their deletion mutants. NADH oxidase-related metabolic pathways were analyzed, further clarifying the function of this enzyme in biofilm formation.

  15. A Cyclic Voltammetry Experiment for the Instrumental Analysis Laboratory.

    Science.gov (United States)

    Baldwin, Richard P.; And Others

    1984-01-01

    Background information and procedures are provided for experiments that illustrate the nature of cyclic voltammetry and its application in the characterization of organic electrode processes. The experiments also demonstrate the concepts of electrochemical reversibility and diffusion-controlled mass transfer. (JN)

  16. Square wave voltammetry at the dropping mercury electrode: Theory

    Science.gov (United States)

    Christie, J.H.; Turner, J.A.; Osteryoung, R.A.

    1977-01-01

    The theoretical aspects of square wave voltammetry at the dropping mercury electrode are presented. The technique involves scanning the entire potential range of interest on a single drop of a DME. Asymmetries in the waveform as well as variations in current measurement parameters are discussed. Indications are that previous uses of the waveform may not have utilized all its capabilities.

  17. Fast Selective Detection of Pyocyanin Using Cyclic Voltammetry

    DEFF Research Database (Denmark)

    Alatraktchi, Fatima AlZahra'a; Andersen, Sandra Breum; Johansen, Helle Krogh;

    2016-01-01

    method to detect pyocyanin in a complex electroactive environment using commercially available electrodes. It is shown that cyclic voltammetry measurements between -1.0 V to 1.0 V reveal a potential detection window of pyocyanin of 0.58-0.82 V that is unaffected by other redox-active interferents...

  18. Fast Selective Detection of Pyocyanin Using Cyclic Voltammetry

    DEFF Research Database (Denmark)

    Al Atraktchi, Fatima Al-Zahraa; Breum Andersen, Sandra; Johansen, Helle Krogh;

    2016-01-01

    method to detect pyocyanin in a complex electroactive environment using commercially available electrodes. It is shown that cyclic voltammetry measurements between –1.0 V to 1.0 V reveal a potential detection window of pyocyanin of 0.58–0.82 V that is unaffected by other redox-active interferents...

  19. Differential cyclic voltammetry for selective and amplified detection

    NARCIS (Netherlands)

    Megen, M.J.J; Odijk, M.; Wiedemair, J.; Olthuis, W.; Berg, van den A.

    2012-01-01

    We propose to combine two existing electrochemical techniques, cyclic voltammetry (CV) and redox cycling (RC), in order to obtain amplified and selective detection of redox active species. This combination is achieved by applying CV waveforms to two electrodes spaced 1.20 mu m apart, with one of the

  20. Chemically Specific Cellular Imaging of Biofilm Formation

    Energy Technology Data Exchange (ETDEWEB)

    Herberg, J L; Schaldach, C; Horn, J; Gjersing, E; Maxwell, R

    2006-02-09

    This document and the accompanying manuscripts summarize the technical accomplishments for our one-year LDRD-ER effort. Biofilm forming microbes have existed on this planet for billions of years and make up 60% of the biological mass on earth. Such microbes exhibit unique biochemical pathways during biofilm formation and play important roles in human health and the environment. Microbial biofilms have been directly implicated in, for example, product contamination, energy losses, and medical infection that cost the loss of human lives and billions of dollars. In no small part due to the lack of detailed understanding, biofilms unfortunately are resistant to control, inhibition, and destruction, either through treatment with antimicrobials or immunological defense mechanisms of the body. Current biofilm research has concentrated on the study of biofilms in the bulk. This is primarily due to the lack of analytical and physical tools to study biofilms non-destructively, in three dimensions, and on the micron or sub-micron scale. This has hindered the development of a clear understanding of either the early stage mechanisms of biofilm growth or the interactions of biofilms with their environment. Enzymatic studies have deduced a biochemical reaction that results in the oxidation of reduced sulfur species with the concomitant reduction of nitrate, a common groundwater pollutant, to dinitrogen gas by the bacterium, Thiobacillus denitrificans (TD). Because of its unique involvement in biologically relevant environmental pathways, TD is scheduled for genome sequencing in the near future by the DOE's Joint Genome Institute and is of interest to DOE's Genomes to Life Program. As our ecosystem is exposed to more and more nitrate contamination large scale livestock and agricultural practices, a further understanding of biofilm formation by organisms that could alleviate these problems is necessary in order to protect out biosphere. However, in order to study this

  1. Interpretation of Cyclic Voltammetry Measurements of Thin Semiconductor Films for Solar Fuel Applications.

    Science.gov (United States)

    Bertoluzzi, Luca; Badia-Bou, Laura; Fabregat-Santiago, Francisco; Gimenez, Sixto; Bisquert, Juan

    2013-04-18

    A simple model is proposed that allows interpretation of the cyclic voltammetry diagrams obtained experimentally for photoactive semiconductors with surface states or catalysts used for fuel production from sunlight. When the system is limited by charge transfer from the traps/catalyst layer and by detrapping, it is shown that only one capacitive peak is observable and is not recoverable in the return voltage scan. If the system is limited only by charge transfer and not by detrapping, two symmetric capacitive peaks can be observed in the cathodic and anodic directions. The model appears as a useful tool for the swift analysis of the electronic processes that limit fuel production.

  2. Sampling phasic dopamine signaling with fast-scan cyclic voltammetry in awake, behaving rats.

    Science.gov (United States)

    Fortin, S M; Cone, J J; Ng-Evans, S; McCutcheon, J E; Roitman, M F

    2015-01-05

    Fast-scan cyclic voltammetry (FSCV) is an electrochemical technique that permits the in vivo measurement of extracellular fluctuations in multiple chemical species. The technique is frequently utilized to sample sub-second (phasic) concentration changes of the neurotransmitter dopamine in awake and behaving rats. Phasic dopamine signaling is implicated in reinforcement, goal-directed behavior, and locomotion, and FSCV has been used to investigate how rapid changes in striatal dopamine concentration contribute to these and other behaviors. This unit describes the instrumentation and construction, implantation, and use of components required to sample and analyze dopamine concentration changes in awake rats with FSCV.

  3. 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.

  4. SinR controls enterotoxin expression in Bacillus thuringiensis biofilms.

    Directory of Open Access Journals (Sweden)

    Annette Fagerlund

    Full Text Available The entomopathogen Bacillus thuringiensis produces dense biofilms under various conditions. Here, we report that the transition phase regulators Spo0A, AbrB and SinR control biofilm formation and swimming motility in B. thuringiensis, just as they control biofilm formation and swarming motility in the closely related saprophyte species B. subtilis. However, microarray analysis indicated that in B. thuringiensis, in contrast to B. subtilis, SinR does not control an eps operon involved in exopolysaccharides production, but regulates genes involved in the biosynthesis of the lipopeptide kurstakin. This lipopeptide is required for biofilm formation and was previously shown to be important for survival in the host cadaver (necrotrophism. Microarray analysis also revealed that the SinR regulon contains genes coding for the Hbl enterotoxin. Transcriptional fusion assays, Western blots and hemolysis assays confirmed that SinR controls Hbl expression, together with PlcR, the main virulence regulator in B. thuringiensis. We show that Hbl is expressed in a sustained way in a small subpopulation of the biofilm, whereas almost all the planktonic population transiently expresses Hbl. The gene coding for SinI, an antagonist of SinR, is expressed in the same biofilm subpopulation as hbl, suggesting that hbl transcription heterogeneity is SinI-dependent. B. thuringiensis and B. cereus are enteric bacteria which possibly form biofilms lining the host intestinal epithelium. Toxins produced in biofilms could therefore be delivered directly to the target tissue.

  5. 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.

  6. Investigation of natural biofilms formed during the production of drinking water from surface water embankment filtration.

    Science.gov (United States)

    Emtiazi, Farahnaz; Schwartz, Thomas; Marten, Silke Mareike; Krolla-Sidenstein, Peter; Obst, Ursula

    2004-03-01

    Populations of bacteria in biofilms from different sites of a drinking water production system were analysed. Polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) analyses revealed changing DNA band patterns, suggesting a population shift during bank filtration and processing at the waterworks. In addition, common DNA bands that were attributed to ubiquitous bacteria were found. Biofilms even developed directly after UV disinfection (1-2m distance). Their DNA band patterns only partly agreed with those of the biofilms from the downstream distribution system. Opportunistic pathogenic bacteria in biofilms were analysed using PCR and Southern blot hybridisation (SBH). Surface water appeared to have a direct influence on the composition of biofilms in the drinking water distribution system. In spite of preceding filtration and UV disinfection, opportunistic pathogens such as atypical mycobacteria and Legionella spp. were found in biofilms of drinking water, and Pseudomonas aeruginosa was detected sporadically. Enterococci were not found in any biofilm. Bacterial cell counts in the biofilms from surface water to drinking water dropped significantly, and esterase and alanine-aminopeptidase activity decreased. beta-glucosidase activity was not found in the biofilms. Contrary to the results for planktonic bacteria, inhibitory effects were not observed in biofilms. This suggested an increased tolerance of biofilm bacteria against toxic compounds.

  7. Biofilm in wound care.

    Science.gov (United States)

    Rajpaul, Kumal

    2015-03-01

    A biofilm can be described as a microbial colony encased in a polysaccharide matrix which can become attached to a wound surface. This can affect the healing potential of chronic wounds due to the production of destructive enzymes and toxins which can promote a chronic inflammatory state within the wound. Biofilms can be polymicrobial and can result in delayed wound healing and chronic wound infection resistant to antibiotics, leading to prolonged hospitalisation for some patients. There appears to be a correlation between biofilms and non-healing in chronic wounds. It is suggested that biofilms are a major player in the chronicity of wounds. They are a complex concept to diagnose and management needs to be multifactorial.

  8. Compaction and relaxation of biofilms

    KAUST Repository

    Valladares Linares, R.

    2015-06-18

    Operation of membrane systems for water treatment can be seriously hampered by biofouling. A better characterization of biofilms in membrane systems and their impact on membrane performance may help to develop effective biofouling control strategies. The objective of this study was to determine the occurrence, extent and timescale of biofilm compaction and relaxation (decompaction), caused by permeate flux variations. The impact of permeate flux changes on biofilm thickness, structure and stiffness was investigated in situ and non-destructively with optical coherence tomography using membrane fouling monitors operated at a constant crossflow velocity of 0.1 m s−1 with permeate production. The permeate flux was varied sequentially from 20 to 60 and back to 20 L m−2 h−1. The study showed that the average biofilm thickness on the membrane decreased after elevating the permeate flux from 20 to 60 L m−2 h−1 while the biofilm thickness increased again after restoring the original flux of 20 L m−2 h−1, indicating the occurrence of biofilm compaction and relaxation. Within a few seconds after the flux change, the biofilm thickness was changed and stabilized, biofilm compaction occurred faster than the relaxation after restoring the original permeate flux. The initial biofilm parameters were not fully reinstated: the biofilm thickness was reduced by 21%, biofilm stiffness had increased and the hydraulic biofilm resistance was elevated by 16%. Biofilm thickness was related to the hydraulic biofilm resistance. Membrane performance losses are related to the biofilm thickness, density and morphology, which are influenced by (variations in) hydraulic conditions. A (temporarily) permeate flux increase caused biofilm compaction, together with membrane performance losses. The impact of biofilms on membrane performance can be influenced (increased and reduced) by operational parameters. The article shows that a (temporary) pressure increase leads to more

  9. Interactions in multispecies biofilms

    DEFF Research Database (Denmark)

    Burmølle, Mette; Ren, Dawei; Bjarnsholt, Thomas;

    2014-01-01

    The recent focus on complex bacterial communities has led to the recognition of interactions across species boundaries. This is particularly pronounced in multispecies biofilms, where synergistic interactions impact the bacterial distribution and overall biomass produced. Importantly, in a number...... of settings, the interactions in a multispecies biofilm affect its overall function, physiology, or surroundings, by resulting in enhanced resistance, virulence, or degradation of pollutants, which is of significant importance to human health and activities. The underlying mechanisms causing these synergistic...

  10. Biofilms in chronic wounds.

    Science.gov (United States)

    James, Garth A; Swogger, Ellen; Wolcott, Randall; Pulcini, Elinor deLancey; Secor, Patrick; Sestrich, Jennifer; Costerton, John W; Stewart, Philip S

    2008-01-01

    Chronic wounds including diabetic foot ulcers, pressure ulcers, and venous leg ulcers are a worldwide health problem. It has been speculated that bacteria colonizing chronic wounds exist as highly persistent biofilm communities. This research examined chronic and acute wounds for biofilms and characterized microorganisms inhabiting these wounds. Chronic wound specimens were obtained from 77 subjects and acute wound specimens were obtained from 16 subjects. Culture data were collected using standard clinical techniques. Light and scanning electron microscopy techniques were used to analyze 50 of the chronic wound specimens and the 16 acute wound specimens. Molecular analyses were performed on the remaining 27 chronic wound specimens using denaturing gradient gel electrophoresis and sequence analysis. Of the 50 chronic wound specimens evaluated by microscopy, 30 were characterized as containing biofilm (60%), whereas only one of the 16 acute wound specimens was characterized as containing biofilm (6%). This was a statistically significant difference (p<0.001). Molecular analyses of chronic wound specimens revealed diverse polymicrobial communities and the presence of bacteria, including strictly anaerobic bacteria, not revealed by culture. Bacterial biofilm prevalence in specimens from chronic wounds relative to acute wounds observed in this study provides evidence that biofilms may be abundant in chronic wounds.

  11. Bacteriophages and Biofilms

    Directory of Open Access Journals (Sweden)

    David R. Harper

    2014-06-01

    Full Text Available Biofilms are an extremely common adaptation, allowing bacteria to colonize hostile environments. They present unique problems for antibiotics and biocides, both due to the nature of the extracellular matrix and to the presence within the biofilm of metabolically inactive persister cells. Such chemicals can be highly effective against planktonic bacterial cells, while being essentially ineffective against biofilms. By contrast, bacteriophages seem to have a greater ability to target this common form of bacterial growth. The high numbers of bacteria present within biofilms actually facilitate the action of bacteriophages by allowing rapid and efficient infection of the host and consequent amplification of the bacteriophage. Bacteriophages also have a number of properties that make biofilms susceptible to their action. They are known to produce (or to be able to induce enzymes that degrade the extracellular matrix. They are also able to infect persister cells, remaining dormant within them, but re-activating when they become metabolically active. Some cultured biofilms also seem better able to support the replication of bacteriophages than comparable planktonic systems. It is perhaps unsurprising that bacteriophages, as the natural predators of bacteria, have the ability to target this common form of bacterial life.

  12. Virulence and pathogenicity of Candida albicans is enhanced in biofilms containing oral bacteria.

    Science.gov (United States)

    Cavalcanti, Yuri Wanderley; Morse, Daniel James; da Silva, Wander José; Del-Bel-Cury, Altair Antoninha; Wei, Xiaoqing; Wilson, Melanie; Milward, Paul; Lewis, Michael; Bradshaw, David; Williams, David Wynne

    2015-01-01

    This study examined the influence of bacteria on the virulence and pathogenicity of candidal biofilms. Mature biofilms (Candida albicans-only, bacteria-only, C. albicans with bacteria) were generated on acrylic and either analysed directly, or used to infect a reconstituted human oral epithelium (RHOE). Analyses included Candida hyphae enumeration and assessment of Candida virulence gene expression. Lactate dehydrogenase (LDH) activity and Candida tissue invasion following biofilm infection of the RHOE were also measured. Candida hyphae were more prevalent (p biofilms also containing bacteria, with genes encoding secreted aspartyl-proteinases (SAP4/SAP6) and hyphal-wall protein (HWP1) up-regulated (p biofilm infections of RHOE. Multi-species infections exhibited higher hyphal proportions (p biofilms promoted Candida virulence, consideration should be given to the bacterial component when managing denture biofilm associated candidoses.

  13. Biofilms and the food industry

    Directory of Open Access Journals (Sweden)

    Nathanon Trachoo

    2003-11-01

    Full Text Available In the past, interest in biofilms was limited to research related to water distribution systems, waste water treatment and dental plaques. Biofilm has become a more popular research topic in many other areas in recent years including food safety. Biofilm formation can compromise the sanitation of food surfaces and environmental surfaces by spreading detached organisms to other areas of processing plants. Unfortunately, these detached organisms are not similar to normal microorganisms suspended in an aquatic environment but are more resistant to several stresses or microbial inactivation including some food preservation methods. Microstructures of biofilms as revealed by different types of microscopic techniques showed that biofilms are highly complex and consist of many symbiotic organisms, some of which are human pathogens. This article reviewed the process of biofilm formation, the significance of biofilms on food or food contact surfaces, their ability to protect foodborne pathogens from environmental stresses and recent methods for the study of biofilms on food contact surfaces.

  14. Thin-film voltammetry and its analytical applications: a review.

    Science.gov (United States)

    Tian, Huihui; Li, Yunchao; Shao, Huibo; Yu, Hua-Zhong

    2015-01-15

    Electrochemical reactions at the interfaces of immiscible electrolyte solutions (ITIES) are of fundamental importance in the fields of chemical, biological and pharmaceutical sciences. Four-electrode cell setup, scanning electrochemical microscopy (SECM) and thin-film voltammetry are the three most frequently used methods for studying the electrochemical processes at these interfaces. The principle, experimental design, advantages and challenges of the three methods are described and compared. The thin-film voltammetry is highlighted for its simplicity in experimental operation and kinetic data analysis. Its versatile analytical applications are discussed in detail, including the study of redox properties of hydrophobic compounds, evaluation of interfacial electron transfer kinetics, synthesis of nanoparticles/nanostructures, and illustration of cross-membrane ion transport phenomena.

  15. Determination of ziram in vegetable samples by square wave voltammetry

    Institute of Scientific and Technical Information of China (English)

    Ping Qiu; Yong Nian Ni

    2008-01-01

    The electrochemical behaviour and electrode reaction mechanism of ziram (zinc-dimethyl dithiocarbamate) on a hanging mercury drop electrode were investigated in Britton-Robinson (B-R) buffer by using cyclic and square wave voltammetry (SWV).Based on these studies a voltammetric method for determination of ziram was developed and applied to determine the ziram in synthetic and spiked vegetable samples,satisfactory results were obtained in both eases.

  16. Differential Pulse Anodic Stripping Voltammetry for Mercury Determination

    OpenAIRE

    2015-01-01

    In the present work voltammetric investigations have been performed on HgCl2 aqueous solutions prepared from a Cz 9024 reagent. Carbon paste electrode (CPE), eriochrome black T modified carbon paste electrode (MCPE/EBT) and KCl 1M as background electrolyte, were involved within the experimental procedures. Cyclic voltammetry (CV) has been performed in order to compare the behaviour of the two electrodes in both K3[Fe(CN)6] and mercury calibration aqueous solution. Differential pulse anodic st...

  17. New Cyclic Voltammetry Method for Examining Phase Transitions: Simulated Results

    OpenAIRE

    2006-01-01

    We propose a new experimental technique for cyclic voltammetry, based on the first-order reversal curve (FORC) method for analysis of systems undergoing hysteresis. The advantages of this electrochemical FORC (EC-FORC) technique are demonstrated by applying it to dynamical models of electrochemical adsorption. The method can not only differentiate between discontinuous and continuous phase transitions, but can also quite accurately recover equilibrium behavior from dynamic analysis of systems...

  18. Surface Catalytic Mechanism in Square-Wave Voltammetry

    OpenAIRE

    Mirceski, Valentin; Gulaboski, Rubin

    2001-01-01

    A pseudo-first-order catalytic mechanism in which both reactant and product of the redox reaction are strongly immobilized on the electrode surface is theoretically analyzed under conditions of square-wave voltammetry (SWV). A mathematical procedure is developed for diffusionless conditions. The relationships between the properties of the voltammetric response and both the kinetic parameters of the redox reaction and the parameters of the excitement signal are studied. The phenomenon...

  19. Oxidation management of white wines using cyclic voltammetry and multivariate process monitoring.

    Science.gov (United States)

    Martins, Rui C; Oliveira, Raquel; Bento, Fatima; Geraldo, Dulce; Lopes, Vitor V; Guedes de Pinho, Paula; Oliveira, Carla M; Silva Ferreira, Antonio C

    2008-12-24

    The development of a fingerprinting strategy capable to evaluate the "oxidation status" of white wines based on cyclic voltammetry is proposed here. It is known that the levels of specific antioxidants and redox mechanisms may be evaluated by cyclic voltammetry. This electrochemical technique was applied on two sets of samples. One group was composed of normal aged white wines and a second group obtained from a white wine forced aging protocol with different oxygen, SO(2), pH, and temperature regimens. A study of antioxidant additions, namely ascorbic acid, was also made in order to establish a statistical link between voltammogram fingerprints and chemical antioxidant substances. It was observed that the oxidation curve presented typical features, which enables sample discrimination according to age, oxygen consumption, and antioxidant additions. In fact, it was possible to place the results into four significant orthogonal directions, compressing 99.8% of nonrandom features. Attempts were made to make voltammogram fingerprinting a tool for monitoring oxidation management. For this purpose, a supervised multivariate control chart was developed using a control sample as reference. When white wines are plotted onto the chart, it is possible to monitor the oxidation status and to diagnose the effects of oxygen regimes and antioxidant activity. Finally, quantification of substances implicated in the oxidation process as reagents (antioxidants) and products (off-flavors) was tried using a supervised algorithmic the partial least square regression analysis. Good correlations (r > 0.93) were observed for ascorbic acid, Folin-Ciocalteu index, total SO(2), methional, and phenylacetaldehyde. These results show that cyclic voltammetry fingerprinting can be used to monitor and diagnose the effects of wine oxidation.

  20. Solution voltammetry of 4 nm magnetite iron oxide nanoparticles.

    Science.gov (United States)

    Roberts, Joseph J P; Westgard, John A; Cooper, Laura M; Murray, Royce W

    2014-07-30

    The voltammetry of solution-dispersed magnetite iron oxide Fe3O4 nanoparticles is described. Their currents are controlled by nanoparticle transport rates, as shown with potential step chronoamperometry and rotated disk voltammetry. In pH 2 citrate buffer with added NaClO4 electrolyte, solution cyclic voltammetry of these nanoparticles (average diameter 4.4 ± 0.9 nm, each containing ca. 30 Fe sites) displays an electrochemically irreversible oxidation with E(PEAK) at ca. +0.52 V and an irreversible reduction with E(PEAK) at ca. +0.2 V vs Ag/AgCl reference electrode. These processes are presumed to correspond to the formal potentials for one-electron oxidation of Fe(II) and reduction of Fe(III) at their different sites in the magnetite nanoparticle structure. The heterogeneous electrode reaction rates of the nanoparticles are very slow, in the 10(-5) cm/s range. The nanoparticles are additionally characterized by a variety of tools, e.g., TEM, UV/vis, and XPS spectroscopies.

  1. Biofilm formation on abiotic surfaces

    DEFF Research Database (Denmark)

    Tang, Lone

    2011-01-01

    Bacteria can attach to any surface in contact with water and proliferate into complex communities enclosed in an adhesive matrix, these communities are called biofilms. The matrix makes the biofilm difficult to remove by physical means, and bacteria in biofilm can survive treatment with many...... to changing the surface hydrophobicity. The influence of surface topography in the biomolecule of great importance for bacterial adhesion...

  2. In-situ biofilm characterization in membrane systems using Optical Coherence Tomography: formation, structure, detachment and impact of flux change.

    Science.gov (United States)

    Dreszer, C; Wexler, A D; Drusová, S; Overdijk, T; Zwijnenburg, A; Flemming, H-C; Kruithof, J C; Vrouwenvelder, J S

    2014-12-15

    Biofouling causes performance loss in spiral wound nanofiltration (NF) and reverse osmosis (RO) membrane operation for process and drinking water production. The development of biofilm formation, structure and detachment was studied in-situ, non-destructively with Optical Coherence Tomography (OCT) in direct relation with the hydraulic biofilm resistance and membrane performance parameters: transmembrane pressure drop (TMP) and feed-channel pressure drop (FCP). The objective was to evaluate the suitability of OCT for biofouling studies, applying a membrane biofouling test cell operated at constant crossflow velocity (0.1 m s(-1)) and permeate flux (20 L m(-2)h(-1)). In time, the biofilm thickness on the membrane increased continuously causing a decline in membrane performance. Local biofilm detachment was observed at the biofilm-membrane interface. A mature biofilm was subjected to permeate flux variation (20 to 60 to 20 L m(-2)h(-1)). An increase in permeate flux caused a decrease in biofilm thickness and an increase in biofilm resistance, indicating biofilm compaction. Restoring the original permeate flux did not completely restore the original biofilm parameters: After elevated flux operation the biofilm thickness was reduced to 75% and the hydraulic resistance increased to 116% of the original values. Therefore, after a temporarily permeate flux increase the impact of the biofilm on membrane performance was stronger. OCT imaging of the biofilm with increased permeate flux revealed that the biofilm became compacted, lost internal voids, and became more dense. Therefore, membrane performance losses were not only related to biofilm thickness but also to the internal biofilm structure, e.g. caused by changes in pressure. Optical Coherence Tomography proved to be a suitable tool for quantitative in-situ biofilm thickness and morphology studies which can be carried out non-destructively and in real-time in transparent membrane biofouling monitors.

  3. In-situ biofilm characterization in membrane systems using Optical Coherence Tomography: Formation, structure, detachment and impact of flux change

    KAUST Repository

    Dreszer, C.

    2014-12-01

    Biofouling causes performance loss in spiral wound nanofiltration (NF) and reverse osmosis (RO) membrane operation for process and drinking water production. The development of biofilm formation, structure and detachment was studied in-situ, non-destructively with Optical Coherence Tomography (OCT) in direct relation with the hydraulic biofilm resistance and membrane performance parameters: transmembrane pressure drop (TMP) and feed-channel pressure drop (FCP). The objective was to evaluate the suitability of OCT for biofouling studies, applying a membrane biofouling test cell operated at constant crossflow velocity (0.1 m s-1) and permeate flux (20 L m-2h-1).In time, the biofilm thickness on the membrane increased continuously causing a decline in membrane performance. Local biofilm detachment was observed at the biofilm-membrane interface. A mature biofilm was subjected to permeate flux variation (20 to 60 to 20 L m-2h-1). An increase in permeate flux caused a decrease in biofilm thickness and an increase in biofilm resistance, indicating biofilm compaction. Restoring the original permeate flux did not completely restore the original biofilm parameters: After elevated flux operation the biofilm thickness was reduced to 75% and the hydraulic resistance increased to 116% of the original values. Therefore, after a temporarily permeate flux increase the impact of the biofilm on membrane performance was stronger. OCT imaging of the biofilm with increased permeate flux revealed that the biofilm became compacted, lost internal voids, and became more dense. Therefore, membrane performance losses were not only related to biofilm thickness but also to the internal biofilm structure, e.g. caused by changes in pressure.Optical Coherence Tomography proved to be a suitable tool for quantitative in-situ biofilm thickness and morphology studies which can be carried out non-destructively and in real-time in transparent membrane biofouling monitors.

  4. In situ molecular imaging of hydrated biofilm in a microfluidic reactor by ToF-SIMS

    Energy Technology Data Exchange (ETDEWEB)

    Hua, Xin; Yu, Xiao-Ying; Wang, Zhaoying; Yang, Li; Liu, Bingwen; Zhu, Zihua; Tucker, Abigail E.; Chrisler, William B.; Hill, Eric A.; Thevuthasan, Suntharampillai; Lin, Yuehe; Liu, Songqin; Marshall, Matthew J.

    2014-02-26

    The first results of using a novel single channel microfluidic reactor to enable Shewanella biofilm growth and in situ characterization using time-of-flight secondary ion mass spectrometry (ToF-SIMS) in the hydrated environment are presented. The new microfluidic interface allows direct probing of the liquid surface using ToF-SIMS, a vacuum surface technique. The detection window is an aperture of 2 m in diameter on a thin silicon nitride (SiN) membrane and it allows direct detection of the liquid surface. Surface tension of the liquid flowing inside the microchannel holds the liquid within the aperture. ToF-SIMS depth profiling was used to drill through the SiN membrane and the biofilm grown on the substrate. In situ 2D imaging of the biofilm in hydrated state was acquired, providing spatial distribution of the chemical compounds in the biofilm system. This data was compared with a medium filled microfluidic reactor devoid of biofilm and dried biofilm samples deposited on clean silicon wafers. Principle Component Analysis (PCA) was used to investigate these observations. Our results show that imaging biofilms in the hydrated environment using ToF-SIMS is possible using the unique microfluidic reactor. Moreover, characteristic biofilm fatty acids fragments were observed in the hydrated biofilm grown in the microfluidic channel, illustrating the advantage of imaging biofilm in its native environment.

  5. Methods for studying biofilm formation: flow cells and confocal laser scanning microscopy

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim; Sternberg, Claus

    2014-01-01

    In this chapter methods for growing and analyzing biofilms under hydrodynamic conditions in flow cells are described. Use of flow cells allows for direct microscopic investigation of biofilm formation. The flow in these chambers is essentially laminar, which means that the biofilms can be grown u......, inoculation of the flow cells, running of the system, confocal laser scanning microscopy and image analysis, and disassembly and cleaning of the system.......In this chapter methods for growing and analyzing biofilms under hydrodynamic conditions in flow cells are described. Use of flow cells allows for direct microscopic investigation of biofilm formation. The flow in these chambers is essentially laminar, which means that the biofilms can be grown...

  6. Molecule Targeting Glucosyltransferase Inhibits Streptococcus mutans Biofilm Formation and Virulence.

    Science.gov (United States)

    Ren, Zhi; Cui, Tao; Zeng, Jumei; Chen, Lulu; Zhang, Wenling; Xu, Xin; Cheng, Lei; Li, Mingyun; Li, Jiyao; Zhou, Xuedong; Li, Yuqing

    2015-10-19

    Dental plaque biofilms are responsible for numerous chronic oral infections and cause a severe health burden. Many of these infections cannot be eliminated, as the bacteria in the biofilms are resistant to the host's immune defenses and antibiotics. There is a critical need to develop new strategies to control biofilm-based infections. Biofilm formation in Streptococcus mutans is promoted by major virulence factors known as glucosyltransferases (Gtfs), which synthesize adhesive extracellular polysaccharides (EPS). The current study was designed to identify novel molecules that target Gtfs, thereby inhibiting S. mutans biofilm formation and having the potential to prevent dental caries. Structure-based virtual screening of approximately 150,000 commercially available compounds against the crystal structure of the glucosyltransferase domain of the GtfC protein from S. mutans resulted in the identification of a quinoxaline derivative, 2-(4-methoxyphenyl)-N-(3-{[2-(4-methoxyphenyl)ethyl]imino}-1,4-dihydro-2-quinoxalinylidene)ethanamine, as a potential Gtf inhibitor. In vitro assays showed that the compound was capable of inhibiting EPS synthesis and biofilm formation in S. mutans by selectively antagonizing Gtfs instead of by killing the bacteria directly. Moreover, the in vivo anti-caries efficacy of the compound was evaluated in a rat model. We found that the compound significantly reduced the incidence and severity of smooth and sulcal-surface caries in vivo with a concomitant reduction in the percentage of S. mutans in the animals' dental plaque (P biofilm formation and the cariogenicity of S. mutans.

  7. Molecular Determinants of Staphylococcal Biofilm Dispersal and Structuring

    Directory of Open Access Journals (Sweden)

    Katherine Y Le

    2014-11-01

    Full Text Available Staphylococci are frequently implicated in human infections, and continue to pose a therapeutic dilemma due to their ability to form deeply seated microbial communities, known as biofilms, on the surfaces of implanted medical devices and host tissues. Biofilm development has been proposed to occur in three stages: 1 attachment, 2 proliferation/structuring, and 3 detachment/dispersal. Although research within the last several decades has implicated multiple molecules in the roles as effectors of staphylococcal biofilm proliferation/structuring and detachment/dispersal, to date, only phenol soluble modulins (PSMs have been consistently demonstrated to serve in this role under both in-vitro and in-vivo settings. PSMs are regulated directly through a density-dependent manner by the accessory gene regulator (Agr system. They disrupt the non-covalent forces holding the biofilm extracellular matrix together, which is necessary for the formation of channels, a process essential for the delivery of nutrients to deeper biofilm layers, and for dispersal/dissemination of clusters of biofilm to distal organs in acute infection. Given their relevance in both acute and chronic biofilm-associated infections, the Agr system and the psm genes hold promise as potential therapeutic targets.

  8. Mannheimia haemolytica biofilm formation on bovine respiratory epithelial cells.

    Science.gov (United States)

    Boukahil, Ismail; Czuprynski, Charles J

    2016-12-25

    Mannheimia haemolytica is the most important bacterial agent associated with the bovine respiratory disease complex (BRDC), which causes worldwide economic losses to the cattle industry. M. haemolytica cells initially colonize the tonsillar crypts in the upper respiratory tract of cattle, from where they can subsequently descend into the lungs to cause disease. Many bacteria exist as biofilms inside their hosts. We hypothesize that M. haemolytica colonization of cattle during its commensal state may include biofilm formation. To begin to assess this possibility, we developed an in vitro system to study biofilm formation directly on bovine respiratory epithelial cells. Using fixed primary bovine bronchial epithelial cells, we observed M. haemolytica biofilm formation after a 48h incubation period at 37°C. Addition of mucin, the main component of mucus present in the upper respiratory tract, decreased M. haemolytica biofilm formation on bovine epithelial cells. We investigated the effects of prior viral infection of the epithelial cells on subsequent biofilm formation by M. haemolytica and found negligible effects. Utilization of this model system will provide new insights into the potential role of biofilm formation by M. haemolytica in the pathogenesis of BRDC.

  9. In vitro characterization of Trichophyton rubrum and T. mentagrophytes biofilms.

    Science.gov (United States)

    Costa-Orlandi, C B; Sardi, J C O; Santos, C T; Fusco-Almeida, A M; Mendes-Giannini, M J S

    2014-01-01

    Dermatophytes are fungi responsible for a disease known as dermatophytosis. Biofilms are sessile microbial communities surrounded by extracellular polymeric substances (EPS) with increased resistance to antimicrobial agents and host defenses. This paper describes, for the first time, the characteristics of Trichophyton rubrum and T. mentagrophytes biofilms. Biofilm formation was analyzed by light microscopy, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) as well as by staining with crystal violet and safranin. Metabolic activity was determined using the XTT reduction assay. Both species were able to form mature biofilms in 72 h. T. rubrum biofilm produced more biomass and EPS and was denser than T. mentagrophytes biofilm. The SEM results demonstrated a coordinated network of hyphae in all directions, embedded within EPS in some areas. Research and characterization of biofilms formed by dermatophytes may contribute to the search of new drugs for the treatment of these mycoses and might inform future revisions with respect to the dose and duration of treatment of currently available antifungals.

  10. Effects of operating conditions on the adhesive strength of Pseudomonas fluorescens biofilms in tubes.

    Science.gov (United States)

    Chen, M J; Zhang, Z; Bott, T R

    2005-06-25

    Understanding the mechanical properties of biofilms, especially the force required to disrupt them and remove them from substrata is very important to development of antibiofouling strategies. In this work, a novel micromanipulation technique with a specially designed T-shaped probe has been developed to serve as an experimental means to measure directly the adhesive strength of biofouling deposits on the surface of a glass test stud. The basic principle of this novel technique is to pull away a whole biofilm accumulated on the surface of a glass test stud with T-shaped probe, and to measure simultaneously the force imposed on the biofilm. The adhesive strength between the biofilms and the surface to which they are attached, is defined as the work per unit area required to remove the biofilms from the surface. The biofouling experiments were performed on an elaborate design of a simulated heat exchanger system. A monoculture of Pseudomonas fluorescens was chosen as the fouling microorganism for the laboratory studies. Results indicate that the adhesive strength of the biofilm was affected by the conditions of operation, such as biofilm age, nutrient concentration, suspended cell concentration, pH, surface roughness of the substratum and fluid velocity. As noted, the effect of fluid velocity on the biofilm adhesive strength seemed to overwhelm other factors. At the same operating conditions, the biofilm adhesive strength increased as the fluid velocity increased within the range of 0.6-1.6m/s. In addition, the flow-related biofilm structures were observed that biofilms generally grew as a more compact pattern at the higher fluid velocity. Apparently, the fluid velocity can affect the biofilm structure, which in turn determines the biofilm adhesive strength. The knowledge of the biofilm adhesive strength with associated influences of the operating conditions may be used to define better cleaning procedures.

  11. Pseudomonas aeruginosa Biofilms

    DEFF Research Database (Denmark)

    Alhede, Maria; Bjarnsholt, Thomas; Givskov, Michael Christian;

    2014-01-01

    biofilms, which protect the aggregated, biopolymer-embedded bacteria from the detrimental actions of antibiotic treatments and host immunity. A key component in the protection against innate immunity is rhamnolipid, which is a quorum sensing (QS)-regulated virulence factor. QS is a cell-to-cell signaling...... mechanism used to coordinate expression of virulence and protection of aggregated biofilm cells. Rhamnolipids are known for their ability to cause hemolysis and have been shown to cause lysis of several cellular components of the human immune system, for example, macrophages and polymorphonuclear leukocytes...

  12. Biofilm Formation by Bacillus cereus Is Influenced by PlcR, a Pleiotropic Regulator

    Science.gov (United States)

    Hsueh, Yi-Huang; Somers, Eileen B.; Lereclus, Didier; Wong, Amy C. Lee

    2006-01-01

    The ΔplcR mutant of Bacillus cereus strain ATCC 14579 developed significantly more biofilm than the wild type and produced increased amounts of biosurfactant. Biosurfactant production is required for biofilm formation and may be directly or indirectly repressed by PlcR, a pleiotropic regulator. Coating polystyrene plates with surfactin, a biosurfactant from Bacillus subtilis, rescued the deficiency in biofilm formation by the wild type. PMID:16820512

  13. Determination of trace amounts of morphine in human plasma by anodic adsorptive stripping differential pulse voltammetry

    Institute of Scientific and Technical Information of China (English)

    Ali Niazi; Ateesa Yazdanipour

    2008-01-01

    New adsorptive anodic differential pulse stripping voltammetry method for the direct determination of morphine at trace levels in human plasma of addicts is proposed.The procedure involves an adsorptive accumulation of morphine on a HMDE,followed by oxidation of adsorbed morphine by voltammetry scan using differential pulse modulation.The optimum conditions for the analysis of morphine are pH 10.5,Eacc of - 100 mV (vs.Ag/AgCl),and tacc of 120 s.The peak current is proportional to the concentration of morphine,and a linear calibration graph is obtained at 0.01-3.10 μg mL-1.A relative standard deviation of 1.06% (n=5) was obtained,and the limit of detection was 3 ng mL- 1.The capability of the method for the analysis of real samples was evaluated by the determination of morphine in spiked human plasma and addicts human plasma with satisfactory results.

  14. Combined characterization of bovine polyhemoglobin microcapsules by UV-Vis absorption spectroscopy and cyclic voltammetry.

    Science.gov (United States)

    Knirsch, Marcos Camargo; Dell'Anno, Filippo; Salerno, Marco; Larosa, Claudio; Polakiewicz, Bronislaw; Eggenhöffner, Roberto; Converti, Attilio

    2017-03-01

    Polyhemoglobin produced from pure bovine hemoglobin by reaction with PEG bis(N-succynimidil succinate) as a cross-linking agent was encapsulated in gelatin and dehydrated by freeze-drying. Free carboxyhemoglobin and polyhemoglobin microcapsules were characterized by UV-Vis spectroscopy in the absorption range 450-650 nm and cyclic voltammetry in the voltage range from -0.8 to 0.6 mV to evaluate the ability to break the bond with carbon monoxide and to study the carrier's affinity for oxygen, respectively. SEM used to observe the shape of cross-linked gelatin-polyhemoglobin microparticles showed a regular distribution of globular shapes, with mean size of ~750 nm, which was ascribed to gelatin. Atomic absorption spectroscopy was also performed to detect iron presence in microparticles. Cyclic voltammetry using an Ag-AgCl electrode highlighted characteristic peaks at around -0.6 mV that were attributed to reversible oxygen bonding with iron in oxy-polyhemoglobin structure. These results suggest this technique as a powerful, direct and alternative method to evaluate the extent of hemoglobin oxygenation.

  15. Assessing principal component regression prediction of neurochemicals detected with fast-scan cyclic voltammetry.

    Science.gov (United States)

    Keithley, Richard B; Wightman, R Mark

    2011-06-07

    Principal component regression is a multivariate data analysis approach routinely used to predict neurochemical concentrations from in vivo fast-scan cyclic voltammetry measurements. This mathematical procedure can rapidly be employed with present day computer programming languages. Here, we evaluate several methods that can be used to evaluate and improve multivariate concentration determination. The cyclic voltammetric representation of the calculated regression vector is shown to be a valuable tool in determining whether the calculated multivariate model is chemically appropriate. The use of Cook's distance successfully identified outliers contained within in vivo fast-scan cyclic voltammetry training sets. This work also presents the first direct interpretation of a residual color plot and demonstrated the effect of peak shifts on predicted dopamine concentrations. Finally, separate analyses of smaller increments of a single continuous measurement could not be concatenated without substantial error in the predicted neurochemical concentrations due to electrode drift. Taken together, these tools allow for the construction of more robust multivariate calibration models and provide the first approach to assess the predictive ability of a procedure that is inherently impossible to validate because of the lack of in vivo standards.

  16. Critical review on biofilm methods.

    Science.gov (United States)

    Azeredo, Joana; Azevedo, Nuno F; Briandet, Romain; Cerca, Nuno; Coenye, Tom; Costa, Ana Rita; Desvaux, Mickaël; Di Bonaventura, Giovanni; Hébraud, Michel; Jaglic, Zoran; Kačániová, Miroslava; Knøchel, Susanne; Lourenço, Anália; Mergulhão, Filipe; Meyer, Rikke Louise; Nychas, George; Simões, Manuel; Tresse, Odile; Sternberg, Claus

    2017-05-01

    Biofilms are widespread in nature and constitute an important strategy implemented by microorganisms to survive in sometimes harsh environmental conditions. They can be beneficial or have a negative impact particularly when formed in industrial settings or on medical devices. As such, research into the formation and elimination of biofilms is important for many disciplines. Several new methodologies have been recently developed for, or adapted to, biofilm studies that have contributed to deeper knowledge on biofilm physiology, structure and composition. In this review, traditional and cutting-edge methods to study biofilm biomass, viability, structure, composition and physiology are addressed. Moreover, as there is a lack of consensus among the diversity of techniques used to grow and study biofilms. This review intends to remedy this, by giving a critical perspective, highlighting the advantages and limitations of several methods. Accordingly, this review aims at helping scientists in finding the most appropriate and up-to-date methods to study their biofilms.

  17. Microbial Biofilms and Chronic Wounds

    Science.gov (United States)

    Omar, Amin; Wright, J. Barry; Schultz, Gregory; Burrell, Robert; Nadworny, Patricia

    2017-01-01

    Background is provided on biofilms, including their formation, tolerance mechanisms, structure, and morphology within the context of chronic wounds. The features of biofilms in chronic wounds are discussed in detail, as is the impact of biofilm on wound chronicity. Difficulties associated with the use of standard susceptibility tests (minimum inhibitory concentrations or MICs) to determine appropriate treatment regimens for, or develop new treatments for use in, chronic wounds are discussed, with alternate test methods specific to biofilms being recommended. Animal models appropriate for evaluating biofilm treatments are also described. Current and potential future therapies for treatment of biofilm-containing chronic wounds, including probiotic therapy, virulence attenuation, biofilm phenotype expression attenuation, immune response suppression, and aggressive debridement combined with antimicrobial dressings, are described. PMID:28272369

  18. Cell-based chip for the detection of anticancer effect on HeLa cells using cyclic voltammetry.

    Science.gov (United States)

    El-Said, Waleed Ahmed; Yea, Cheol-Heon; Kim, Hyunhee; Oh, Byung-Keun; Choi, Jeong-Woo

    2009-01-01

    HeLa cells directly immobilized on gold-patterned silicon substrate were used to assess the biological toxicity of anticancer drugs (hydroxyurea and cyclophosphamide). Immobilization of HeLa cells was confirmed by optical microscopy, and cell growth, viability and drug-related toxicity were examined by cyclic voltammetry and potentiometric stripping analysis. The voltammetric behaviors of HeLa cells displayed a quasi-reversible pattern with the peak current exhibiting a linear relationship with cell number. The attached living cells were exposed to different concentrations of hydroxyurea and cyclophosphamide as anticancer drugs, which induced the change of cyclic voltammetry current peak. As the exposed concentration of anticancer drugs was increased, the change of current peak was increased, which indicates the decrease of cell viability. Trypan Blue dyeing was performed to confirm the results of the effect of anticancer drugs on the cell viability which was obtained from cyclic voltammetry assay. The proposed direct cell immobilization method technique can be applied to the fabrication of cell chip for diagnosis, drug detection, and on-site monitoring.

  19. Esoteric communiqué amid microbes in an oral biofilm

    Directory of Open Access Journals (Sweden)

    Harpreet Singh Grover

    2013-01-01

    Full Text Available Dental biofilms are complex and multispecies ecosystems, and its formation requires coordinated chemical signaling between different micro-organisms present in the oral cavity. During the initial stages of its formation, planktonic bacterial cells directly attach to surfaces of the oral cavity or indirectly bind to other bacterial cells. This binding occurs through co-aggregation, which is critical for the temporary retention of bacteria on dental surfaces as well as bacterial colonization. It is during this colonization that the micro-organisms are able to interact with each other. In general, interspecies interactions involve communication, typically via quorum sensing, and metabolic cooperation or competition. Interactions among species within a biofilm can be antagonistic, such as competition over nutrients and growth inhibition, or synergistic. In this review, we discuss these important interactions among oral bacteria within the dental biofilm communities and novel therapies that could inhibit pathogenic micro-organisms and disrupt biofilm.

  20. The Root Canal Biofilm

    NARCIS (Netherlands)

    Sluis, van der L.W.M.; Boutsioukis, C.; Jiang, L.M.; Macedo, R.; Verhaagen, B.; Versluis, M.; Chávez de Paz, E.; Sedgley, C.M.; Kishen, A.

    2015-01-01

    The aims of root canal irrigation are the chemical dissolution or disruption and the mechanical detachment of pulp tissue, dentin debris and smear layer (instrumentation products), microorganisms (planktonic or biofilm), and their products from the root canal wall, their removal out of the root cana

  1. Vibrio cholerae biofilm growth program and architecture revealed by single-cell live imaging.

    Science.gov (United States)

    Yan, Jing; Sharo, Andrew G; Stone, Howard A; Wingreen, Ned S; Bassler, Bonnie L

    2016-09-01

    Biofilms are surface-associated bacterial communities that are crucial in nature and during infection. Despite extensive work to identify biofilm components and to discover how they are regulated, little is known about biofilm structure at the level of individual cells. Here, we use state-of-the-art microscopy techniques to enable live single-cell resolution imaging of a Vibrio cholerae biofilm as it develops from one single founder cell to a mature biofilm of 10,000 cells, and to discover the forces underpinning the architectural evolution. Mutagenesis, matrix labeling, and simulations demonstrate that surface adhesion-mediated compression causes V. cholerae biofilms to transition from a 2D branched morphology to a dense, ordered 3D cluster. We discover that directional proliferation of rod-shaped bacteria plays a dominant role in shaping the biofilm architecture in V. cholerae biofilms, and this growth pattern is controlled by a single gene, rbmA Competition analyses reveal that the dense growth mode has the advantage of providing the biofilm with superior mechanical properties. Our single-cell technology can broadly link genes to biofilm fine structure and provides a route to assessing cell-to-cell heterogeneity in response to external stimuli.

  2. In situ generation and detection of methyl radical by voltammetry

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    In persulfate-acetate, dimethyl sulfoxide or tert-butyl alcohol systems, in situ generation and detection of methyl free radical are realized with voltammetry. It includes the following successive processes. The persulfate S2O82- is polarographically reduced via one-electron addition to sulfate radical SO4-., the SO4-. Initiates chain reaction with acetate, dimethyl sulfoxide, or tert-butyl alcohol on the electrode surface to produce a methyl radical, and one-electron reduction of the methyl radical yields its polarographic reduction wave. In comparison with the known techniques such as ultraviolet radiolysis coupling with electron spin resonance, etc., the proposed method is simple, sensitive and selective.

  3. The effects of silver nanoparticles on intact wastewater biofilms

    Directory of Open Access Journals (Sweden)

    Zhiya eSheng

    2015-07-01

    Full Text Available Silver nanoparticles (Ag-NPs have strong antibacterial properties, which may adversely affect biological wastewater treatment processes. To determine the overall effect, intact biofilm samples were collected from the rotating biological contactor (RBC at the local wastewater treatment plant and treated with 200 mg Ag/L Ag-NPs for 24 h. The biofilm uptake of Ag-NPs was monitored with transmission electron microscopy (TEM. Forty-five min after Ag-NP application, Ag-NPs were seen in the biofilm extracellular polymeric substances (EPS. After 24 h, Ag-NPs had entered certain microbial cells, while other cells contained no observable Ag-NPs. Some cells were dying after the uptake of Ag-NPs. However, there was no significant reduction in cultivable bacteria in the biofilms, based on heterotrophic plate counts (HPC. While this may indicate that wastewater biofilms are highly resistant to Ag-NPs, the HPC represents only a small portion of the total microbial population. To further investigate the effects of Ag-NPs, a GeoChip microarray was used to directly detect changes in the functional gene structure of the microbial community in the biofilm. A clear decrease (34.6% decrease in gene number in gene diversity was evident in the GeoChip analysis. However, the complete loss of any specific gene was rare. Some gene families present in both treated and untreated biofilms. However, this doesn’t necessarily mean that there was no change in these families. Signal intensity decreased in certain variants in each family while other variants increased to compensate the effects of Ag-NPs. The results indicate that Ag-NP treatment decreased microbial community diversity but did not significantly affect the microbial community function. This provides direct evidence for the functional redundancy of microbial community in engineered ecosystems such as wastewater biofilms.

  4. Biofilm-associated infection: the hidden face of cerebrospinal fluid shunt malfunction.

    Science.gov (United States)

    Mounier, Roman; Kapandji, Natacha; Birnbaum, Ron; Cook, Fabrice; Rodriguez, Cristophe; Nebbad, Bibba; Lobo, David; Dhonneur, Gilles

    2016-12-01

    Diagnosis of cerebrospinal fluid (CSF) shunt infection is difficult. Growing evidence links this pattern to biofilm-associated infections (BAI). Biofilm may explain the indolent development of the infection, and the poor efficiency of traditional microbiologic methods. We report the case of a patient admitted for hydrocephalus associated to CSF shunt malfunction. None of the clinical, serum, or CSF laboratory findings were in favor of an infectious process. Only scanning electron microscopy (SEM) revealed the presence of biofilm. Hence, despite a broad CSF shunt infection definition, some infections could remain undiagnosed by the traditional approach. This study is the first to provide some direct evidence for bacterial biofilm-associated CSF shunt infection.

  5. A sticky situation: untangling the transcriptional network controlling biofilm development in Candida albicans.

    Science.gov (United States)

    Fox, Emily P; Nobile, Clarissa J

    2012-01-01

    Candida albicans is a commensal microorganism of the human microbiome; it is also the most prevalent fungal pathogen of humans. Many infections caused by C. albicans are a direct consequence of its proclivity to form biofilms--resilient, surface-associated communities of cells where individual cells acquire specialized properties that are distinct from those observed in suspension cultures. We recently identified the transcriptional network that orchestrates the formation of biofilms in C. albicans. These results set the stage for understanding how biofilms are formed and, once formed, how the specialized properties of biofilms are elaborated. This information will provide new insight for understanding biofilms in more detail and may lead to improvements in preventing and treating biofilm-based infections in the future.

  6. Dissolved inorganic carbon enhanced growth, nutrient uptake, and lipid accumulation in wastewater grown microalgal biofilms.

    Science.gov (United States)

    Kesaano, Maureen; Gardner, Robert D; Moll, Karen; Lauchnor, Ellen; Gerlach, Robin; Peyton, Brent M; Sims, Ronald C

    2015-03-01

    Microalgal biofilms grown to evaluate potential nutrient removal options for wastewaters and feedstock for biofuels production were studied to determine the influence of bicarbonate amendment on their growth, nutrient uptake capacity, and lipid accumulation after nitrogen starvation. No significant differences in growth rates, nutrient removal, or lipid accumulation were observed in the algal biofilms with or without bicarbonate amendment. The biofilms possibly did not experience carbon-limited conditions because of the large reservoir of dissolved inorganic carbon in the medium. However, an increase in photosynthetic rates was observed in algal biofilms amended with bicarbonate. The influence of bicarbonate on photosynthetic and respiration rates was especially noticeable in biofilms that experienced nitrogen stress. Medium nitrogen depletion was not a suitable stimulant for lipid production in the algal biofilms and as such, focus should be directed toward optimizing growth and biomass productivities to compensate for the low lipid yields and increase nutrient uptake.

  7. Study on Hydro-Alcoholic Extract Effect of Pomegranate Peel on Pseudomonas aeruginosa Biofilm Formation

    Directory of Open Access Journals (Sweden)

    R. Habibipour

    2015-10-01

    Full Text Available Introduction & Objective: Microorganisms form biomass as biofilm in response to many factors, in order to adapt to hostile extracellular environments and biocides. Using different herbal compounds are of those strategies to deal with biofilm. It has been proved that plants extracts such as pomegranate, raspberry and chamomile essential oils have anti-biofilm effects. This study aimed to evaluate the effect of different concentrations of black peel pomegranate ex-tract on Pseudomonas aeruginosa biofilm formation. Materials & Methods: In this experimental research the anti-biofilm effect, reducing the amount of biofilm formation and growth kinetics of Pseudomonas aeruginosa in different treatments was measured by microtiter and plate colorimetric crystal violet method. Biofilm formation was also examined using a microscope. Statistical analysis of data obtained from the reading of the ELISA was performed using SPSS software, P value 0.05. Results: Findings of this study showed that bacteria cannot form any biofilm in first 6 hours of incubation, in all treatments. The amount of biofilm formation after 12 hours in 0.01 and 0.05 g/ mL treatments were medium. Among treatments, after 18 and 24 hours of incubation 0.001 g/ mL concentration of pomegranate peel extract had medium and strong inhibitory effect on biofilm formation, respectively. Conclusion: Results of this study showed that biofilm formation and biofilm reduction percent-age is directly related to the duration of exposure of bacteria that could be due to the different phases of growth. Growth kinetics study also revealed that in the majority of treatments the growth was incremental up to about 15 hours and decrement afterwards due to the effective-ness of different treatments. After 18 hours, treatments have greatest influence on biofilm formation. The foregoing has been fully confirmed by the results of microscopic slides. (Sci J Hamadan Univ Med Sci 2015; 22 (3: 195-202

  8. Voltage biasing, cyclic voltammetry, & electrical impedance spectroscopy for neural interfaces.

    Science.gov (United States)

    Wilks, Seth J; Richner, Tom J; Brodnick, Sarah K; Kipke, Daryl R; Williams, Justin C; Otto, Kevin J

    2012-02-24

    Electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV) measure properties of the electrode-tissue interface without additional invasive procedures, and can be used to monitor electrode performance over the long term. EIS measures electrical impedance at multiple frequencies, and increases in impedance indicate increased glial scar formation around the device, while cyclic voltammetry measures the charge carrying capacity of the electrode, and indicates how charge is transferred at different voltage levels. As implanted electrodes age, EIS and CV data change, and electrode sites that previously recorded spiking neurons often exhibit significantly lower efficacy for neural recording. The application of a brief voltage pulse to implanted electrode arrays, known as rejuvenation, can bring back spiking activity on otherwise silent electrode sites for a period of time. Rejuvenation alters EIS and CV, and can be monitored by these complementary methods. Typically, EIS is measured daily as an indication of the tissue response at the electrode site. If spikes are absent in a channel that previously had spikes, then CV is used to determine the charge carrying capacity of the electrode site, and rejuvenation can be applied to improve the interface efficacy. CV and EIS are then repeated to check the changes at the electrode-tissue interface, and neural recordings are collected. The overall goal of rejuvenation is to extend the functional lifetime of implanted arrays.

  9. Thin film voltammetry of metabolic enzymes in rat liver microsomes

    Science.gov (United States)

    Krishnan, Sadagopan; Rusling, James F.

    2007-01-01

    We report herein thin film voltammetry and kinetics of electron transfer for redox proteins in rat liver microsomes for the first time. Films were made layer-by-layer from liver microsomes and polycations on pyrolytic graphite electrodes. Cyclic voltammograms were chemically reversible with a midpoint potential of −0.48 V vs SCE at 0.1 V s−1 in pH 7.0 phosphate buffer. Reduction peak potentials shifted negative at higher scan rates, and oxidation-reduction peak current ratios were ∼1 consistent with non-ideal quasireversible thin film voltammetry. Analysis of oxidation-reduction peak separations gave an average apparent surface electron transfer rate constant of 30 s−1. Absence of significant electrocatalytic reduction of O2 or H2O2 and lack of shift in midpoint potential when CO is added that indicates lack of an iron heme cofactor suggest that peaks can be attributed to oxidoreductases present in the microsomes rather than cytochrome P450 enzymes. PMID:18037986

  10. Strategies for low detection limit measurements with cyclic voltammetry.

    Science.gov (United States)

    Wiedemann, D J; Kawagoe, K T; Kennedy, R T; Ciolkowski, E L; Wightman, R M

    1991-12-15

    Cyclic voltammetry of Nafion-coated, carbon-fiber electrodes is used to detect trace concentrations of dopamine, both in a flow injection apparatus and in the brain of an anaesthetized rat. To improve signal-to-noise ratios, the sources of noise during cyclic voltammetry have been determined and strategies have been developed to decrease the noise. With the potentiostat employed, the measured noise is comparable to that expected for Johnson noise from the feedback resistor of the current transducer. Additional noise arises from the waveform generator employed and, in some cases, line noise. Line noise is discriminated against by starting each cyclic voltammogram either in phase or 180 degrees out of phase with the line frequency. When used in vivo, additional noise also arises from the physiological activity of the animal. Detection limits are found to closely correspond to those predicted on the basis of simulation of the voltammetric shape and the measured noise. Detection limits are improved by the use of appropriate analog and digital filtering, ensemble averaging, and appropriate timing of repetitive cyclic voltammograms. The combined use of these techniques enables the in vivo detection of approximately 100 nM of dopamine with a signal-to-noise ratio of 25.

  11. Differential Pulse Anodic Stripping Voltammetry for Mercury Determination

    Directory of Open Access Journals (Sweden)

    Vereștiuc Paul C.

    2015-07-01

    Full Text Available In the present work voltammetric investigations have been performed on HgCl2 aqueous solutions prepared from a Cz 9024 reagent. Carbon paste electrode (CPE, eriochrome black T modified carbon paste electrode (MCPE/EBT and KCl 1M as background electrolyte, were involved within the experimental procedures. Cyclic voltammetry (CV has been performed in order to compare the behaviour of the two electrodes in both K3[Fe(CN6] and mercury calibration aqueous solution. Differential pulse anodic stripping voltammetry (DPASV was used to determine the most suitable parameters for mercury determination. All experiments were performed at 25 ± 1 ℃, using an electrochemical cell with three-electrodes connected to an Autolab PG STAT 302N (Metrohm-Autolab potentiostat that is equipped with Nova 1.11 software. The measured potential values were generated by using the silver chloride electrode (AgClE as reference and a platinum wire electrode as auxiliary. A series of time depending equations for the pre-concentration and concentration steps were established, with the observation that a higher sensitivity can be obtained while increasing the pre-concentration time. DPASV were drawn using the CPE in 11.16 % coriander, as mercury complex, the voltamograms signals indicating mercury oxidation, with signal intensity increasing in time.

  12. Biofilm susceptibility to metal toxicity.

    Science.gov (United States)

    Harrison, Joe J; Ceri, Howard; Stremick, Carol A; Turner, Raymond J

    2004-12-01

    This study compared bacterial biofilm and planktonic cell susceptibility to metal toxicity by evaluating the minimum inhibitory concentration (MIC), the planktonic minimum bactericidal concentration (MBC), and minimum biofilm eradication concentration (MBEC) using the MBEC device. In total, 17 metal cations and oxyanions, chosen to represent groups VIB to VIA of the periodic table, were each tested on biofilm and planktonic cultures of Escherichia coli JM109, Staphylococcus aureus ATCC 29213, and Pseudomonas aeruginosa ATCC 27853. In contrast to control antibiotic assays, where biofilm cultures were 2 to 64 times less susceptible to killing than logarithmically growing planktonic bacteria, metal compounds killed planktonic and biofilm cultures at the same concentration in the vast majority of combinations. Our data indicate that, under the conditions reported, growth in a biofilm does not provide resistance to bacteria against killing by metal cations or oxyanions.

  13. Critical review on biofilm methods

    DEFF Research Database (Denmark)

    Azeredo, Joana; F. Azevedo, Nuno; Briandet, Romain;

    2017-01-01

    Biofilms are widespread in nature and constitute an important strategy implemented by microorganisms to survive in sometimes harsh environmental conditions. They can be beneficial or have a negative impact particularly when formed in industrial settings or on medical devices. As such, research...... into the formation and elimination of biofilms is important for many disciplines. Several new methodologies have been recently developed for, or adapted to, biofilm studies that have contributed to deeper knowledge on biofilm physiology, structure and composition. In this review, traditional and cutting-edge methods...... to study biofilm biomass, viability, structure, composition and physiology are addressed. Moreover, as there is a lack of consensus among the diversity of techniques used to grow and study biofilms. This review intends to remedy this, by giving a critical perspective, highlighting the advantages...

  14. Critical review on biofilm methods

    DEFF Research Database (Denmark)

    Azeredo, Joana; F. Azevedo, Nuno; Briandet, Romain;

    2017-01-01

    into the formation and elimination of biofilms is important for many disciplines. Several new methodologies have been recently developed for, or adapted to, biofilm studies that have contributed to deeper knowledge on biofilm physiology, structure and composition. In this review, traditional and cutting-edge methods...... to study biofilm biomass, viability, structure, composition and physiology are addressed. Moreover, as there is a lack of consensus among the diversity of techniques used to grow and study biofilms. This review intends to remedy this, by giving a critical perspective, highlighting the advantages...... and limitations of several methods. Accordingly, this review aims at helping scientists in finding the most appropriate and up-to-date methods to study their biofilms....

  15. Biofilms: a developing microscopic community

    Directory of Open Access Journals (Sweden)

    Rivera Sandra Patricia

    2004-09-01

    Full Text Available Biofilms are microbial communities composed by different microbiota embebbed in a special adaptive environment. These communities show different characteristics such as heterogeneity, diversity in microenvironments, capacity to resist antimicrobial therapy and ability to allow bacterial communication. These characteristics convert them in complex organizations that are difficult to eradicate in their own environment. In the man, biofilms are associated to a great number of slow-development infectious processes which greatly difficulties their eradication. In the industry and environment, biofilms are centered in processes known as biofouling and bioremediation. The former is the contamination of a system due to the microbial activity of a biofilm. The latter uses biofilms to improve the conditions of a contaminated system. The study of biofilms is a new and exciting field which is constantly evolving and whose implications in medicine and industry would have important repercussions for the humankind.

  16. Lead migration from toys by anodic stripping voltammetry using a bismuth film electrode.

    Science.gov (United States)

    Leal, M Fernanda C; Catarino, Rita I L; Pimenta, Adriana M; Souto, M Renata S; Afonso, Christelle S; Fernandes, Ana F Q

    2016-09-02

    Metals may be released from toys via saliva during mouthing, via sweat during dermal contact, or via gastric and intestinal fluids after partial or whole ingestion. In this study, we determined the lead migration from toys bought on the Portuguese market for children below 3 years of age. The lead migration was performed according to the European Committee for Standardization EN 71-3, which proposes a 2-hour migration test that simulates human gastric conditions. The voltammetric determination of migrated lead was performed by anodic stripping voltammetry (ASV) at a bismuth film electrode (BiFE). For all the analyzed toys, the values of migrated lead did not exceed the limits imposed by the European Committee for Standardization EN 71-3 (90 mg kg(-1)) and by the EU Directive 2009/48/EC (13.5 mg kg(-1)) on the safety of toys.

  17. Evaluation of Salmonella biofilm cell transfer from contact surfaces to beef products

    Science.gov (United States)

    Introduction: Meat contamination by Salmonella enterica is a serious food safety concern. One common transmission route that leads to cross contamination in meat plants is bacteria transfer from biofilms on contact surfaces to meat products via direct contact. Many factors could affect biofilm tra...

  18. Investigation of Streptococcus mutans biofilm growth on modified Au(111)-surfaces using AFM and electrochemistry

    DEFF Research Database (Denmark)

    Hu, Yifan; Zhang, Jingdong; Ulstrup, Jens

    2011-01-01

    Biofilms of the bacterium Streptococcus mutans constitute perhaps the most important direct cause of human dental caries formation. We have studied S. mutans biofilm formation and properties on Au(111)-surfaces modified by self-assembled molecular monolayers (SAMs) of different thiol-based molecu...

  19. Determination of Platinum Metals in Carbonaceous Mineral Raw Materials by Stripping Voltammetry

    OpenAIRE

    Kolpakova, Nina Alexandrovna; Oskina, Yuliya Aleksandrovna; Dyachenko, Elena Nikolaevna; Pshenichkin, Anatoliy Yakovlevich

    2015-01-01

    The paper considers the possibility of determining platinum metals in mineral raw materials by stripping voltammetry on a graphite electrode modified by metals. Stripping voltammetry method is characterized by low determination limit, wide intervals of determined content and high sensitivity. As a result of the research the conditions for the determination of gold, platinum and palladium by stripping voltammetry have been selected. The comparison of the results of gold, palladium and platinum...

  20. Boron doped diamond sensor for sensitive determination of metronidazole: Mechanistic and analytical study by cyclic voltammetry and square wave voltammetry.

    Science.gov (United States)

    Ammar, Hafedh Belhadj; Brahim, Mabrouk Ben; Abdelhédi, Ridha; Samet, Youssef

    2016-02-01

    The performance of boron-doped diamond (BDD) electrode for the detection of metronidazole (MTZ) as the most important drug of the group of 5-nitroimidazole was proven using cyclic voltammetry (CV) and square wave voltammetry (SWV) techniques. A comparison study between BDD, glassy carbon and silver electrodes on the electrochemical response was carried out. The process is pH-dependent. In neutral and alkaline media, one irreversible reduction peak related to the hydroxylamine derivative formation was registered, involving a total of four electrons. In acidic medium, a prepeak appears probably related to the adsorption affinity of hydroxylamine at the electrode surface. The BDD electrode showed higher sensitivity and reproducibility analytical response, compared with the other electrodes. The higher reduction peak current was registered at pH11. Under optimal conditions, a linear analytical curve was obtained for the MTZ concentration in the range of 0.2-4.2μmolL(-1), with a detection limit of 0.065μmolL(-1).

  1. Boron doped diamond sensor for sensitive determination of metronidazole: Mechanistic and analytical study by cyclic voltammetry and square wave voltammetry

    Energy Technology Data Exchange (ETDEWEB)

    Ammar, Hafedh Belhadj, E-mail: hbelhadjammar@yahoo.fr; Brahim, Mabrouk Ben; Abdelhédi, Ridha; Samet, Youssef

    2016-02-01

    The performance of boron-doped diamond (BDD) electrode for the detection of metronidazole (MTZ) as the most important drug of the group of 5-nitroimidazole was proven using cyclic voltammetry (CV) and square wave voltammetry (SWV) techniques. A comparison study between BDD, glassy carbon and silver electrodes on the electrochemical response was carried out. The process is pH-dependent. In neutral and alkaline media, one irreversible reduction peak related to the hydroxylamine derivative formation was registered, involving a total of four electrons. In acidic medium, a prepeak appears probably related to the adsorption affinity of hydroxylamine at the electrode surface. The BDD electrode showed higher sensitivity and reproducibility analytical response, compared with the other electrodes. The higher reduction peak current was registered at pH 11. Under optimal conditions, a linear analytical curve was obtained for the MTZ concentration in the range of 0.2–4.2 μmol L{sup −1}, with a detection limit of 0.065 μmol L{sup −1}. - Highlights: • SWV for the determination of MTZ • Boron-doped diamond as a new electrochemical sensor • Simple and rapid detection of MTZ • Efficiency of BDD for sensitive determination of MTZ.

  2. Biofilm and Dental Biomaterials

    Directory of Open Access Journals (Sweden)

    Marit Øilo

    2015-05-01

    Full Text Available All treatment involving the use of biomaterials in the body can affect the host in positive or negative ways. The microbiological environment in the oral cavity is affected by the composition and shape of the biomaterials used for oral restorations. This may impair the patients’ oral health and sometimes their general health as well. Many factors determine the composition of the microbiota and the formation of biofilm in relation to biomaterials such as, surface roughness, surface energy and chemical composition, This paper aims to give an overview of the scientific literature regarding the association between the chemical, mechanical and physical properties of dental biomaterials and oral biofilm formation, with emphasis on current research and future perspectives.

  3. Effects of warming on stream biofilm organic matter use capabilities.

    Science.gov (United States)

    Ylla, Irene; Canhoto, Cristina; Romaní, Anna M

    2014-07-01

    The understanding of ecosystem responses to changing environmental conditions is becoming increasingly relevant in the context of global warming. Microbial biofilm communities in streams play a key role in organic matter cycling which might be modulated by shifts in flowing water temperature. In this study, we performed an experiment at the Candal stream (Portugal) longitudinally divided into two reaches: a control half and an experimental half where water temperature was 3 °C above that of the basal stream water. Biofilm colonization was monitored during 42 days in the two stream halves. Changes in biofilm function (extracellular enzyme activities and carbon substrate utilization profiles) as well as chlorophyll a and prokaryote densities were analyzed. The biofilm in the experimental half showed a higher capacity to decompose cellulose, hemicellulose, lignin, and peptidic compounds. Total leucine-aminopeptidase, cellobiohydrolase and β-xylosidase showed a respective 93, 66, and 61% increase in activity over the control; much higher than would be predicted by only the direct temperature physical effect. In contrast, phosphatase and lipase activity showed the lowest sensitivity to temperature. The biofilms from the experimental half also showed a distinct functional fingerprint and higher carbon usage diversity and richness, especially due to a wider use of polymers and carbohydrates. The changes in the biofilm functional capabilities might be indirectly affected by the higher prokaryote and chlorophyll density measured in the biofilm of the experimental half. The present study provides evidence that a realistic stream temperature increase by 3 °C changes the biofilm metabolism to a greater decomposition of polymeric complex compounds and peptides but lower decomposition of lipids. This might affect stream organic matter cycling and the transfer of carbon to higher trophic levels.

  4. Examination of Rapid Dopamine Dynamics with Fast Scan Cyclic Voltammetry During Intra-oral Tastant Administration in Awake Rats.

    Science.gov (United States)

    Wickham, Robert J; Park, Jinwoo; Nunes, Eric J; Addy, Nii A

    2015-08-12

    Rapid, phasic dopamine (DA) release in the mammalian brain plays a critical role in reward processing, reinforcement learning, and motivational control. Fast scan cyclic voltammetry (FSCV) is an electrochemical technique with high spatial and temporal (sub-second) resolution that has been utilized to examine phasic DA release in several types of preparations. In vitro experiments in single-cells and brain slices and in vivo experiments in anesthetized rodents have been used to identify mechanisms that mediate dopamine release and uptake under normal conditions and in disease models. Over the last 20 years, in vivo FSCV experiments in awake, freely moving rodents have also provided insight of dopaminergic mechanisms in reward processing and reward learning. One major advantage of the awake, freely moving preparation is the ability to examine rapid DA fluctuations that are time-locked to specific behavioral events or to reward or cue presentation. However, one limitation of combined behavior and voltammetry experiments is the difficulty of dissociating DA effects that are specific to primary rewarding or aversive stimuli from co-occurring DA fluctuations that mediate reward-directed or other motor behaviors. Here, we describe a combined method using in vivo FSCV and intra-oral infusion in an awake rat to directly investigate DA responses to oral tastants. In these experiments, oral tastants are infused directly to the palate of the rat--bypassing reward-directed behavior and voluntary drinking behavior--allowing for direct examination of DA responses to tastant stimuli.

  5. Exoelectrogenic bacterium phylogenetically related to Citrobacter freundii, isolated from anodic biofilm of a microbial fuel cell.

    Science.gov (United States)

    Huang, Jianjian; Zhu, Nengwu; Cao, Yanlan; Peng, Yue; Wu, Pingxiao; Dong, Wenhao

    2015-02-01

    An electrogenic bacterium, named Citrobacter freundii Z7, was isolated from the anodic biofilm of microbial fuel cell (MFC) inoculated with aerobic sewage sludge. Cyclic voltammetry (CV) analysis exhibited that the strain Z7 had relatively high electrochemical activity. When the strain Z7 was inoculated into MFC, the maximum power density can reach 204.5 mW/m(2) using citrate as electron donor. Series of substrates including glucose, glycerol, lactose, sucrose, and rhammose could be utilized to generate power. CV tests and the addition of anode solution as well as AQDS experiments indicated that the strain Z7 might transfer electrons indirectly via secreted mediators.

  6. Biofilm architecture in a novel pressurized biofilm reactor.

    Science.gov (United States)

    Jiang, Wei; Xia, Siqing; Duan, Liang; Hermanowicz, Slawomir W

    2015-01-01

    A novel pure-oxygen pressurized biofilm reactor was operated at different organic loading, mechanical shear and hydrodynamic conditions to understand the relationships between biofilm architecture and its operation. The ultimate goal was to improve the performance of the biofilm reactor. The biofilm was labeled with seven stains and observed with confocal laser scanning microscopy. Unusual biofilm architecture of a ribbon embedded between two surfaces with very few points of attachment was observed. As organic loading increased, the biofilm morphology changed from a moderately rough layer into a locally smoother biomass with significant bulging protuberances, although the chemical oxygen demand (COD) removal efficiency remained unchanged at about 75%. At higher organic loadings, biofilms contained a larger fraction of active cells distributed uniformly within a proteinaceous matrix with decreasing polysaccharide content. Higher hydrodynamic shear in combination with high organic loading resulted in the collapse of biofilm structure and a substantial decrease in reactor performance (a COD removal of 16%). Moreover, the important role of proteins for the spatial distribution of active cells was demonstrated quantitatively.

  7. Protein film voltammetry and co-factor electron transfer dynamics in spinach photosystem II core complex.

    Science.gov (United States)

    Zhang, Yun; Magdaong, Nikki; Frank, Harry A; Rusling, James F

    2014-05-01

    Direct protein film voltammetry (PFV) was used to investigate the redox properties of the photosystem II (PSII) core complex from spinach. The complex was isolated using an improved protocol not used previously for PFV. The PSII core complex had high oxygen-evolving capacity and was incorporated into thin lipid and polyion films. Three well-defined reversible pairs of reduction and oxidation voltammetry peaks were observed at 4 °C in the dark. Results were similar in both types of films, indicating that the environment of the PSII-bound cofactors was not influenced by film type. Based on comparison with various control samples including Mn-depleted PSII, peaks were assigned to chlorophyll a (Chl a) (Em = -0.47 V, all vs. NHE, at pH 6), quinones (-0.12 V), and the manganese (Mn) cluster (Em = 0.18 V). PFV of purified iron heme protein cytochrome b-559 (Cyt b-559), a component of PSII, gave a partly reversible peak pair at 0.004 V that did not have a potential similar to any peaks observed from the intact PSII core complex. The closest peak in PSII to 0.004 V is the 0.18 V peak that was found to be associated with a two-electron process, and thus is inconsistent with iron heme protein voltammetry. The -0.47 V peak had a peak potential and peak potential-pH dependence similar to that found for purified Chl a incorporated into DMPC films. The midpoint potentials reported here may differ to various extents from previously reported redox titration data due to the influence of electrode double-layer effects. Heterogeneous electron transfer (hET) rate constants were estimated by theoretical fitting and digital simulations for the -0.47 and 0.18 V peaks. Data for the Chl a peaks were best fit to a one-electron model, while the peak assigned to the Mn cluster was best fit by a two-electron/one-proton model.

  8. Determination of Pesticide Ethion by Linear Sweep Stripping Voltammetry

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The presence of the pesticide ethion was determined using an electroanalytical method, i. e. , linear sweep stripping voltammetry at a hanging mercury drop electrode in the Britton-Robbinson buffer with pH = 8. 36, based on its reduction peak observed at - 690 mV following its alkaline hydrolysis. The experimental parameters, namely, the preconcentration potential, the preconcentration time, and the scan rate were investigated and -300 mV (vs. Ag/AgCl), 270 s, and 100 mV/s were selected as the optimal values, respectively. The linear relationship between the peak current and the concentration was found to be in the range of 0. 02-0. 16 mg/L, with a detection limit of 0.0087 mg/L. The proposed method was applied to the determination of ethion in spiked vegetable and fruit samples via a preextraction with anhydrous ethanol.

  9. Nanoalloy electrocatalysis: simulating cyclic voltammetry from configurational thermodynamics with adsorbates.

    Science.gov (United States)

    Wang, Lin-Lin; Tan, Teck L; Johnson, Duane D

    2015-11-14

    We simulate the adsorption isotherms for alloyed nanoparticles (nanoalloys) with adsorbates to determine cyclic voltammetry (CV) during electrocatalysis. The effect of alloying on nanoparticle adsorption isotherms is provided by a hybrid-ensemble Monte Carlo simulation that uses the cluster expansion method extended to non-exchangeable coupled lattices for nanoalloys with adsorbates. Exemplified here for the hydrogen evolution reaction, a 2-dimensional CV is mapped for Pd-Pt nanoalloys as a function of both electrochemical potential and the global Pt composition, and shows a highly non-linear alloying effect on CV. Detailed features in CV arise from the interplay among the H-adsorption in multiple sites that is closely correlated with alloy configurations, which are in turn affected by the H-coverage. The origins of specific features in CV curves are assigned. The method provides a more complete means to design nanoalloys for electrocatalysis.

  10. Underpotential deposition and anodic stripping voltammetry at mesoporous microelectrodes.

    Science.gov (United States)

    Sanchez, Pablo Lozano; Elliott, Joanne M

    2005-05-01

    Using the technique of liquid crystal templating a series of high surface area mesoporous platinum microelectrodes was fabricated. The underpotential deposition of metal ions at such electrodes was found to be similar to that at conventional platinum electrodes. The phenomena of underpotential deposition, in combination with the intrinsic properties of mesoporous microelectrodes (i.e. a high surface area and efficient mass transport) was exploited for the purpose of anodic stripping voltammetry. In particular the underpotential deposition of Ag(+), Pb(2+) and Cu(2+) ions was investigated and it was found that mesoporous microelectrodes were able to quantify the concentration of ions in solution down to the ppb range. The overall behaviour of the mesoporous electrodes was found to be superior to that of conventional microelectrodes and the effects of interference by surfactants were minimal.

  11. Study of quinones reactions with wine nucleophiles by cyclic voltammetry.

    Science.gov (United States)

    Oliveira, Carla M; Barros, António S; Ferreira, António C S; Silva, Artur M S

    2016-11-15

    Quinones are electrophilic species which can react with various nucleophiles, like wine antioxidants, such as sulfur dioxide or ascorbic acid, thiols, amino acids, and numerous polyphenols. These reactions are very important in wine aging because they mediate oxygen reactions during both production and bottle aging phases. In this work, the major challenge was to determine the interaction between ortho-quinones and wine nucleophiles (amino acids, thiols, and the antioxidants SO2 and ascorbic acid), by cyclic voltammetry. Wine-model solutions with gallic acid, caffeic acid, or (+)-catechin and nucleophilic compounds were used. To understand the effect of nucleophilic addition in wine, a white wine with the same added nucleophiles was also analysed. Cyclic voltammograms were taken with glassy carbon electrode or screen-printed carbon electrodes, respectively, for wine-model and white wines solutions, in the absence and in the presence of nucleophiles. A nucleophilic order profile related to the cathodic current intensity decrease was observed.

  12. Computer-controlled instrumentation for fast voltammetry at ultramicroelectrodes

    Science.gov (United States)

    Pospíšil, L.; Fiedler, J.; Fanelli, N.

    2000-04-01

    In this article we describe an electrochemical experimental setup for cyclic voltammetry operating from low (1 V/s) to very high (200 kV/s) scan rates. The system is designed to achieve an acceptable cost and to be user friendly for nonexperts. The instrumentation is based on a commercial arbitrary wave form generator, a digital oscilloscope, a general purpose interface bus interface to a PC, and a laboratory-built potentiostat. There are no complicated manual operations, instead the software creates dialog with an operator to control commands for the instruments. Requirements for the wave form synthesis and the data acquisition are given. Various data-smoothing methods for treating 8-bit fast scan data are evaluated. The performance was tested on the oxidation of ferrocene in acetonitrile. The estimated value of the standard heterogeneous rate constant, k0=0.8±0.1 cms-1, is in reasonable agreement with the most recent reports from other laboratories.

  13. Comparison of the in vitro activity of five antimicrobial drugs on Staphylococcus pseudintermedius and Staphylococcus aureus biofilms

    Directory of Open Access Journals (Sweden)

    Aude A Ferran

    2016-08-01

    Full Text Available Resistance in canine pathogenic staphylococci is necessitating re-evaluation of the current antimicrobial treatments especially for biofilm-associated infections. Long, repeated treatments are often required to control such infections due to the tolerance of bacteria within the biofilm. To comply with the goal of better antibiotic stewardship in veterinary medicine, the efficacies of the available drugs need to be directly assessed on bacterial biofilms.We compared the activities of amoxicillin, cefalexin, clindamycin, doxycycline and marbofloxacin on in vitro biofilms of Staphylococcus pseudintermedius and Staphylococcus aureus. Exposure of biofilms for 15 hours to maximum concentrations of the antibiotics achievable in canine plasma only reduced biofilm bacteria by 0.5 to 2.0 log10 CFU, compared to the control, except for marbofloxacin which reduced S. aureus biofilms by 5.4 log10 CFU. Two-antibiotic combinations did not improve, and even decreased, bacterial killing. In comparison, 5 min-exposure to 2 % chlorhexidine reduced biofilms of the 2 tested strains by 4 log10 CFU. Our results showed that S. pseudintermedius biofilm, unlike S. aureus biofilm, was highly tolerant to all the drugs tested, consistent with the treatment failures observed in practice. Under our conditions, the use of topical chlorhexidine would probably be the best currently available strategy to reduce S. pseudintermedius biofilm.

  14. Cloud Point Extraction for Electroanalysis: Anodic Stripping Voltammetry of Cadmium.

    Science.gov (United States)

    Rusinek, Cory A; Bange, Adam; Papautsky, Ian; Heineman, William R

    2015-06-16

    Cloud point extraction (CPE) is a well-established technique for the preconcentration of hydrophobic species from water without the use of organic solvents. Subsequent analysis is then typically performed via atomic absorption spectroscopy (AAS), UV-vis spectroscopy, or high performance liquid chromatography (HPLC). However, the suitability of CPE for electroanalytical methods such as stripping voltammetry has not been reported. We demonstrate the use of CPE for electroanalysis using the determination of cadmium (Cd(2+)) by anodic stripping voltammetry (ASV). Rather than using the chelating agents which are commonly used in CPE to form a hydrophobic, extractable metal complex, we used iodide and sulfuric acid to neutralize the charge on Cd(2+) to form an extractable ion pair. This offers good selectivity for Cd(2+) as no interferences were observed from other heavy metal ions. Triton X-114 was chosen as the surfactant for the extraction because its cloud point temperature is near room temperature (22-25 °C). Bare glassy carbon (GC), bismuth-coated glassy carbon (Bi-GC), and mercury-coated glassy carbon (Hg-GC) electrodes were compared for the CPE-ASV. A detection limit for Cd(2+) of 1.7 nM (0.2 ppb) was obtained with the Hg-GC electrode. ASV with CPE gave a 20x decrease (4.0 ppb) in the detection limit compared to ASV without CPE. The suitability of this procedure for the analysis of tap and river water samples was demonstrated. This simple, versatile, environmentally friendly, and cost-effective extraction method is potentially applicable to a wide variety of transition metals and organic compounds that are amenable to detection by electroanalytical methods.

  15. Staphylococcus aureus biofilms: recent developments in biofilm dispersal.

    Science.gov (United States)

    Lister, Jessica L; Horswill, Alexander R

    2014-01-01

    Staphylococcus aureus is a major cause of nosocomial and community-acquired infections and represents a significant burden on the healthcare system. S. aureus attachment to medical implants and host tissue, and the establishment of a mature biofilm, play an important role in the persistence of chronic infections. The formation of a biofilm, and encasement of cells in a polymer-based matrix, decreases the susceptibility to antimicrobials and immune defenses, making these infections difficult to eradicate. During infection, dispersal of cells from the biofilm can result in spread to secondary sites and worsening of the infection. In this review, we discuss the current understanding of the pathways behind biofilm dispersal in S. aureus, with a focus on enzymatic and newly described broad-spectrum dispersal mechanisms. Additionally, we explore potential applications of dispersal in the treatment of biofilm-mediated infections.

  16. Microbial composition and antibiotic resistance of biofilms recovered from endotracheal tubes of mechanically ventilated patients.

    Science.gov (United States)

    Vandecandelaere, Ilse; Coenye, Tom

    2015-01-01

    In critically ill patients, breathing is impaired and mechanical ventilation, using an endotracheal tube (ET) connected to a ventilator, is necessary. Although mechanical ventilation is a life-saving procedure, it is not without risk. Because of several reasons, a biofilm often forms at the distal end of the ET and this biofilm is a persistent source of bacteria which can infect the lungs, causing ventilator-associated pneumonia (VAP). There is a link between the microbial flora of ET biofilms and the microorganisms involved in the onset of VAP. Culture dependent and independent techniques were already used to identify the microbial flora of ET biofilms and also, the antibiotic resistance of microorganisms obtained from ET biofilms was determined. The ESKAPE pathogens play a dominant role in the onset of VAP and these organisms were frequently identified in ET biofilms. Also, antibiotic resistant microorganisms were frequently present in ET biofilms. Members of the normal oral flora were also identified in ET biofilms but it is thought that these organisms initiate ET biofilm formation and are not directly involved in the development of VAP.

  17. A three-step method for analysing bacterial biofilm formation under continuous medium flow.

    Science.gov (United States)

    Schmutzler, Karolin; Schmid, Andreas; Buehler, Katja

    2015-07-01

    For the investigation and comparison of microbial biofilms, a variety of analytical methods have been established, all focusing on different growth stages and application areas of biofilms. In this study, a novel quantitative assay for analysing biofilm maturation under the influence of continuous flow conditions was developed using the interesting biocatalyst Pseudomonas taiwanensis VLB120. In contrast to other tubular-based assay systems, this novel assay format delivers three readouts using a single setup in a total assay time of 40 h. It combines morphotype analysis of biofilm colonies with the direct quantification of biofilm biomass and pellicle formation on an air/liquid interphase. Applying the Tube-Assay, the impact of the second messenger cyclic diguanylate on biofilm formation of P. taiwanensis VLB120 was investigated. To this end, 41 deletions of genes encoding for protein homologues to diguanylate cyclase and phosphodiesterase were generated in the genome of P. taiwanensis VLB120. Subsequently, the biofilm formation of the resulting mutants was analysed using the Tube-Assay. In more than 60 % of the mutants, a significantly altered biofilm formation as compared to the parent strain was detected. Furthermore, the potential of the proposed Tube-Assay was validated by investigating the biofilms of several other bacterial species.

  18. A comparison of effects of broad-spectrum antibiotics and biosurfactants on established bacterial biofilms.

    Science.gov (United States)

    Quinn, Gerry A; Maloy, Aaron P; Banat, Malik M; Banat, Ibrahim M

    2013-11-01

    Current antibiofilm solutions based on planktonic bacterial physiology have limited efficacy in clinical and occasionally environmental settings. This has prompted a search for suitable alternatives to conventional therapies. This study compares the inhibitory properties of two biological surfactants (rhamnolipids and a plant-derived surfactant) against a selection of broad-spectrum antibiotics (ampicillin, chloramphenicol and kanamycin). Testing was carried out on a range of bacterial physiologies from planktonic and mixed bacterial biofilms. Rhamnolipids (Rhs) have been extensively characterised for their role in the development of biofilms and inhibition of planktonic bacteria. However, there are limited direct comparisons with antimicrobial substances on established biofilms comprising single or mixed bacterial strains. Baseline measurements of inhibitory activity using planktonic bacterial assays established that broad-spectrum antibiotics were 500 times more effective at inhibiting bacterial growth than either Rhs or plant surfactants. Conversely, Rhs and plant biosurfactants reduced biofilm biomass of established single bacterial biofilms by 74-88 and 74-98 %, respectively. Only kanamycin showed activity against biofilms of Bacillus subtilis and Staphylococcus aureus. Broad-spectrum antibiotics were also ineffective against a complex biofilm of marine bacteria; however, Rhs and plant biosurfactants reduced biofilm biomass by 69 and 42 %, respectively. These data suggest that Rhs and plant-derived surfactants may have an important role in the inhibition of complex biofilms.

  19. Imaging of bacterial multicellular behaviour in biofilms in liquid by atmospheric scanning electron microscopy

    Science.gov (United States)

    Sugimoto, Shinya; Okuda, Ken-ichi; Miyakawa, Reina; Sato, Mari; Arita-Morioka, Ken-ichi; Chiba, Akio; Yamanaka, Kunitoshi; Ogura, Teru; Mizunoe, Yoshimitsu; Sato, Chikara

    2016-01-01

    Biofilms are complex communities of microbes that attach to biotic or abiotic surfaces causing chronic infectious diseases. Within a biofilm, microbes are embedded in a self-produced soft extracellular matrix (ECM), which protects them from the host immune system and antibiotics. The nanoscale visualisation of delicate biofilms in liquid is challenging. Here, we develop atmospheric scanning electron microscopy (ASEM) to visualise Gram-positive and -negative bacterial biofilms immersed in aqueous solution. Biofilms cultured on electron-transparent film were directly imaged from below using the inverted SEM, allowing the formation of the region near the substrate to be studied at high resolution. We visualised intercellular nanostructures and the exocytosis of membrane vesicles, and linked the latter to the trafficking of cargos, including cytoplasmic proteins and the toxins hemolysin and coagulase. A thick dendritic nanotube network was observed between microbes, suggesting multicellular communication in biofilms. A universal immuno-labelling system was developed for biofilms and tested on various examples, including S. aureus biofilms. In the ECM, fine DNA and protein networks were visualised and the precise distribution of protein complexes was determined (e.g., straight curli, flagella, and excreted cytoplasmic molecular chaperones). Our observations provide structural insights into bacteria-substratum interactions, biofilm development and the internal microbe community. PMID:27180609

  20. DNA-microarrays identification of Streptococcus mutans genes associated with biofilm thickness

    Directory of Open Access Journals (Sweden)

    Feldman Mark

    2008-12-01

    Full Text Available Abstract Background A biofilm is a complex community of microorganisms that develop on surfaces in diverse environments. The thickness of the biofilm plays a crucial role in the physiology of the immobilized bacteria. The most cariogenic bacteria, mutans streptococci, are common inhabitants of a dental biofilm community. In this study, DNA-microarray analysis was used to identify differentially expressed genes associated with the thickness of S. mutans biofilms. Results Comparative transcriptome analyses indicated that expression of 29 genes was differentially altered in 400- vs. 100-microns depth and 39 genes in 200- vs. 100-microns biofilms. Only 10 S. mutans genes showed differential expression in both 400- vs. 100-microns and 200- vs. 100-microns biofilms. All of these genes were upregulated. As sucrose is a predominant factor in oral biofilm development, its influence was evaluated on selected genes expression in the various depths of biofilms. The presence of sucrose did not noticeably change the regulation of these genes in 400- vs. 100-microns and/or 200- vs. 100-microns biofilms tested by real-time RT-PCR. Furthermore, we analyzed the expression profile of selected biofilm thickness associated genes in the luxS- mutant strain. The expression of those genes was not radically changed in the mutant strain compared to wild-type bacteria in planktonic condition. Only slight downregulation was recorded in SMU.2146c, SMU.574, SMU.609, and SMU.987 genes expression in luxS- bacteria in biofilm vs. planktonic environments. Conclusion These findings reveal genes associated with the thickness of biofilms of S. mutans. Expression of these genes is apparently not regulated directly by luxS and is not necessarily influenced by the presence of sucrose in the growth media.

  1. Effects of patterned topography on biofilm formation

    Science.gov (United States)

    Vasudevan, Ravikumar

    2011-12-01

    was observed between the patterned topography tested. Another potential strategy for biofilm control through patterned topography is based on the design of robust non-wetting surfaces with undercut feature geometries, characterized by 1) breakthrough pressure and 2) triple phase contact line model. It was found that height and presence of undercut had statistically significant effects, directly proportional to breakthrough pressures, whereas extent of undercut did not. A predictive triple phase contact line model was also developed. (Full text of this dissertation may be available via the University of Florida Libraries web site. Please check http://www.uflib.ufl.edu/etd.html)

  2. Experimental evolution in biofilm populations.

    Science.gov (United States)

    Steenackers, Hans P; Parijs, Ilse; Foster, Kevin R; Vanderleyden, Jozef

    2016-05-01

    Biofilms are a major form of microbial life in which cells form dense surface associated communities that can persist for many generations. The long-life of biofilm communities means that they can be strongly shaped by evolutionary processes. Here, we review the experimental study of evolution in biofilm communities. We first provide an overview of the different experimental models used to study biofilm evolution and their associated advantages and disadvantages. We then illustrate the vast amount of diversification observed during biofilm evolution, and we discuss (i) potential ecological and evolutionary processes behind the observed diversification, (ii) recent insights into the genetics of adaptive diversification, (iii) the striking degree of parallelism between evolution experiments and real-life biofilms and (iv) potential consequences of diversification. In the second part, we discuss the insights provided by evolution experiments in how biofilm growth and structure can promote cooperative phenotypes. Overall, our analysis points to an important role of biofilm diversification and cooperation in bacterial survival and productivity. Deeper understanding of both processes is of key importance to design improved antimicrobial strategies and diagnostic techniques.

  3. Interaction of Nanoparticles with Biofilms

    Science.gov (United States)

    In this work we have studied the interaction and adsorption of engineered nanoparticles such as TiO2, ZnO, CeO2 , and carbon nanotubes with biofilms. Biofilm is an extracellular polymeric substance coating comprised of living material and it is an aggregation of bacteria, algae, ...

  4. Experimental evolution in biofilm populations

    Science.gov (United States)

    Steenackers, Hans P.; Parijs, Ilse; Foster, Kevin R.; Vanderleyden, Jozef

    2016-01-01

    Biofilms are a major form of microbial life in which cells form dense surface associated communities that can persist for many generations. The long-life of biofilm communities means that they can be strongly shaped by evolutionary processes. Here, we review the experimental study of evolution in biofilm communities. We first provide an overview of the different experimental models used to study biofilm evolution and their associated advantages and disadvantages. We then illustrate the vast amount of diversification observed during biofilm evolution, and we discuss (i) potential ecological and evolutionary processes behind the observed diversification, (ii) recent insights into the genetics of adaptive diversification, (iii) the striking degree of parallelism between evolution experiments and real-life biofilms and (iv) potential consequences of diversification. In the second part, we discuss the insights provided by evolution experiments in how biofilm growth and structure can promote cooperative phenotypes. Overall, our analysis points to an important role of biofilm diversification and cooperation in bacterial survival and productivity. Deeper understanding of both processes is of key importance to design improved antimicrobial strategies and diagnostic techniques. PMID:26895713

  5. Microalgal biofilms for wastewater treatment

    NARCIS (Netherlands)

    Boelee, N.C.

    2013-01-01

    The objective of this thesis was to explore the possibilities of using microalgal biofilms for the treatment of municipal wastewater, with a focus on the post-treatment of municipal wastewater effluent. The potential of microalgal biofilms for wastewater treatment was first investigated using a scen

  6. Microbial ecology of phototrophic biofilms

    NARCIS (Netherlands)

    Roeselers, G.

    2007-01-01

    Biofilms are layered structures of microbial cells and an extracellular matrix of polymeric substances, associated with surfaces and interfaces. Biofilms trap nutrients for growth of the enclosed microbial community and help prevent detachment of cells from surfaces in flowing systems. Phototrophic

  7. Microbial biofilms are able to destroy hydroxyapatite in the absence of host immunity in vitro

    Science.gov (United States)

    Junka, Adam Feliks; Szymczyk, Patrycja; Smutnicka, Danuta; Kos, Marcin; Smolina, Iryna; Bartoszewicz, Marzenna; Chlebus, Edward; Turniak, Michal; Sedghizadeh, Parish P.

    2014-01-01

    Introduction It is widely thought that inflammation and osteoclastogenesis result in hydroxyapatite (HA) resorption and sequestra formation during osseous infections, and microbial biofilm pathogens induce the inflammatory destruction of HA. We hypothesized that biofilms associated with infectious bone disease can directly resorb HA in the absence of host inflammation or osteoclastogenesis. Therefore, we developed an in vitro model to test this hypothesis. Materials and Methods Customized HA discs were manufactured as a substrate for growing clinically relevant biofilm pathogens. Single-species biofilms of S.mutans, S.aureus, P.aeruginosa and C.albicans, and mixed-species biofilms of C.albicans + S.mutans were incubated on HA discs for 72 hours to grow mature biofilms. Three different non-biofilm control groups were also established for testing. HA discs were then evaluated by means of scanning electron microscopy, micro-CT metrotomography, x-ray spectroscopy and confocal microscopy with planimetric analysis. Additionally, quantitative cultures and pH assessment were performed. ANOVA was used to test for significance between treatment and control groups. Results All investigated biofilms were able to cause significant (P<0.05) and morphologically characteristic alterations in HA structure as compared to controls. The highest number of alterations observed was caused by mixed biofilms of C.albicans + S.mutans. S. mutans biofilm incubated in medium with additional sucrose content was the most detrimental to HA surfaces among single-species biofilms. Conclusion These findings suggest that direct microbial resorption of bone is possible in addition to immune-mediated destruction, which has important translational implications for the pathogenesis of chronic bone infections and for targeted antimicrobial therapeutics. PMID:25544303

  8. Antibiotic tolerance and microbial biofilms

    DEFF Research Database (Denmark)

    Folkesson, Anders

    Increased tolerance to antimicrobial agents is thought to be an important feature of microbes growing in biofilms. We study the dynamics of antibiotic action within hydrodynamic flow chamber biofilms of Escherichia coli and Pseudomonas aeruginosa using isogenic mutants and fluorescent gene...... expression reporters and we address the question of how biofilm organization affects antibiotic susceptibility. The dynamics of microbial killing is monitored by viable count determination, and confocal laser microscopy. Our work shows that the apparent increased antibiotic tolerance is due to the formation...... of antibiotic tolerant subpopulations within the biofilm. The formation of these subpopulations is highly variable and dependent on the antibiotic used, the biofilm structural organization and the induction of specific tolerance mechanisms....

  9. 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...

  10. Bacterial interactions in dental biofilm.

    Science.gov (United States)

    Huang, Ruijie; Li, Mingyun; Gregory, Richard L

    2011-01-01

    Biofilms are masses of microorganisms that bind to and multiply on a solid surface, typically with a fluid bathing the microbes. The microorganisms that are not attached but are free floating in an aqueous environment are termed planktonic cells. Traditionally, microbiology research has addressed results from planktonic bacterial cells. However, many recent studies have indicated that biofilms are the preferred form of growth of most microbes and particularly those of a pathogenic nature. Biofilms on animal hosts have significantly increased resistance to various antimicrobials compared to planktonic cells. These microbial communities form microcolonies that interact with each other using very sophisticated communication methods (i.e., quorum-sensing). The development of unique microbiological tools to detect and assess the various biofilms around us is a tremendously important focus of research in many laboratories. In the present review, we discuss the major biofilm mechanisms and the interactions among oral bacteria.

  11. Voltammetry and Electrocatalysis of Achrornobacter Xylosoxidans Copper Nitrite Reductase on Functionalized Au(111)-Electrode Surfaces

    DEFF Research Database (Denmark)

    Welinder, Anna C.; Zhang, Jingdong; Hansen, Allan G.;

    2007-01-01

    A long-standing issue in protein film voltammetry (PFV), particularly electrocatalytic voltammetry of redox enzyme monolayers, is the variability of protein adsorption modes, reflected in distributions of catalytic activity of the adsorbed protein/enzyme molecules. Use of well-defined, atomically...

  12. 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

  13. Bacterial biofilms: prokaryotic adventures in multicellularity

    DEFF Research Database (Denmark)

    Webb, J.S.; Givskov, Michael Christian; Kjelleberg, S.

    2003-01-01

    The development of bacterial biofilms includes both the initial social behavior of undifferentiated cells, as well as cell death and differentiation in the mature biofilm, and displays several striking similarities with higher organisms. Recent advances in the field provide new insight...... into differentiation and cell death events in bacterial biofilm development and propose that biofilms have an unexpected level of multicellularity....

  14. Impact of Hydrodynamics on Oral Biofilm Strength

    NARCIS (Netherlands)

    Paramonova, E.; Kalmykowa, O. J.; van der Mei, H. C.; Busscher, H. J.; Sharma, P. K.

    2009-01-01

    Mechanical removal of oral biofilms is ubiquitously accepted as the best way to prevent caries and periodontal diseases. Removal effectiveness strongly depends on biofilm strength. To investigate the influence of hydrodynamics on oral biofilm strength, we grew single- and multi-species biofilms of S

  15. Oral Biofilm Architecture on Natural Teeth

    NARCIS (Netherlands)

    Zijnge, Vincent; van Leeuwen, M. Barbara M.; Degener, John E.; Abbas, Frank; Thurnheer, Thomas; Gmuer, Rudolf; Harmsen, Hermie J. M.

    2010-01-01

    Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and acces

  16. Chemically Specific Cellular Imaging of Biofilm Formation

    Energy Technology Data Exchange (ETDEWEB)

    Herberg, J L; Schaldach, C; Horn, J; Gjersing, E; Maxwell, R

    2006-02-09

    This document and the accompanying manuscripts summarize the technical accomplishments for our one-year LDRD-ER effort. Biofilm forming microbes have existed on this planet for billions of years and make up 60% of the biological mass on earth. Such microbes exhibit unique biochemical pathways during biofilm formation and play important roles in human health and the environment. Microbial biofilms have been directly implicated in, for example, product contamination, energy losses, and medical infection that cost the loss of human lives and billions of dollars. In no small part due to the lack of detailed understanding, biofilms unfortunately are resistant to control, inhibition, and destruction, either through treatment with antimicrobials or immunological defense mechanisms of the body. Current biofilm research has concentrated on the study of biofilms in the bulk. This is primarily due to the lack of analytical and physical tools to study biofilms non-destructively, in three dimensions, and on the micron or sub-micron scale. This has hindered the development of a clear understanding of either the early stage mechanisms of biofilm growth or the interactions of biofilms with their environment. Enzymatic studies have deduced a biochemical reaction that results in the oxidation of reduced sulfur species with the concomitant reduction of nitrate, a common groundwater pollutant, to dinitrogen gas by the bacterium, Thiobacillus denitrificans (TD). Because of its unique involvement in biologically relevant environmental pathways, TD is scheduled for genome sequencing in the near future by the DOE's Joint Genome Institute and is of interest to DOE's Genomes to Life Program. As our ecosystem is exposed to more and more nitrate contamination large scale livestock and agricultural practices, a further understanding of biofilm formation by organisms that could alleviate these problems is necessary in order to protect out biosphere. However, in order to study this

  17. Real-time measurements of the redox states of c-type cytochromes in electroactive biofilms: a confocal resonance Raman Microscopy study.

    Directory of Open Access Journals (Sweden)

    Bernardino Virdis

    Full Text Available Confocal Resonance Raman Microscopy (CRRM was used to probe variations of redox state of c-type cytochromes embedded in living mixed-culture electroactive biofilms exposed to different electrode polarizations, under potentiostatic and potentiodynamic conditions. In the absence of the metabolic substrate acetate, the redox state of cytochromes followed the application of reducing and oxidizing electrode potentials. Real-time monitoring of the redox state of cytochromes during cyclic voltammetry (CV in a potential window where cytochromes reduction occurs, evidenced a measurable time delay between the oxidation of redox cofactors probed by CV at the electrode interface, and oxidation of distal cytochromes probed by CRRM. This delay was used to tentatively estimate the diffusivity of electrons through the biofilm. In the presence of acetate, the resonance Raman spectra of young (10 days, j = 208 ± 49 µA cm(-2 and mature (57 days, j = 267 ± 73 µA cm(-2 biofilms show that cytochromes remained oxidized homogeneously even at layers as far as 70 µm from the electrode, implying the existence of slow metabolic kinetics that do not result in the formation of a redox gradient inside the biofilm during anode respiration. However, old biofilms (80 days, j = 190 ± 37 µA cm(-2 with thickness above 100 µm were characterized by reduced catalytic activity compared to the previous developing stages. The cytochromes in these biofilm were mainly in the reduced redox state, showing that only aged mixed-culture biofilms accumulate electrons during anode respiration. These results differ substantially from recent observations in pure Geobacter sulfurreducens electroactive biofilms, in which accumulation of reduced cytochromes is already observed in thinner biofilms, thus suggesting different bottlenecks in current production for mixed-culture and G. sulfurreducens biofilms.

  18. A brief review: Ultrafast electron diffractive voltammetry: General formalism and applications

    CERN Document Server

    Chang, Kiseok; Tao, Zhensheng; Han, Tzong-Ru T; Ruan, Chong-Yu

    2013-01-01

    We present a general formalism of ultrafast diffractive voltammetry approach as a contact-free tool to investigate the ultrafast surface charge dynamics in nanostructured interfaces. As case studies, the photoinduced surface charging processes in oxidized silicon surface and the hot electron dynamics in nanoparticle-decorated interface are examined based on the diffractive voltammetry framework. We identify that the charge redistribution processes appear on the surface, sub-surface, and vacuum levels when driven by intense femtosecond laser pulses. To elucidate the voltammetry contribution from different sources, we perform controlled experiments using shadow imaging techniques and N-particle simulations to aid the investigation of the photovoltage dynamics in the presence of pho- toemission. We show that voltammetry contribution associated with photoemission has a long decay tail and plays a more visible role in the nanosecond timescale, whereas the ultrafast voltammetry are dominated by local charge transfe...

  19. An ecological perspective of Listeria monocytogenes biofilms in food processing facilities.

    Science.gov (United States)

    Valderrama, Wladir B; Cutter, Catherine N

    2013-01-01

    Listeria monocytogenes can enter the food chain at virtually any point. However, food processing environments seem to be of particular importance. From an ecological point of view, food processing facilities are microbial habitats that are constantly disturbed by cleaning and sanitizing procedures. Although L. monocytogenes is considered ubiquitous in nature, it is important to recognize that not all L. monocytogenes strains appear to be equally distributed; the distribution of the organism seems to be related to certain habitats. Currently, no direct evidence exists that L. monocytogenes-associated biofilms have played a role in food contamination or foodborne outbreaks, likely because biofilm isolation and identification are not part of an outbreak investigation, or the definition of biofilm is unclear. Because L. monocytogenes is known to colonize surfaces, we suggest that contamination patterns may be studied in the context of how biofilm formation is influenced by the environment within food processing facilities. In this review, direct and indirect epidemiological and phenotypic evidence of lineage-related biofilm formation capacity to specific ecological niches will be discussed. A critical view on the development of the biofilm concept, focused on the practical implications, strengths, and weaknesses of the current definitions also is discussed. The idea that biofilm formation may be an alternative surrogate for microbial fitness is proposed. Furthermore, current research on the influence of environmental factors on biofilm formation is discussed.

  20. Environmental factors that shape biofilm formation.

    Science.gov (United States)

    Toyofuku, Masanori; Inaba, Tomohiro; Kiyokawa, Tatsunori; Obana, Nozomu; Yawata, Yutaka; Nomura, Nobuhiko

    2015-01-01

    Cells respond to the environment and alter gene expression. Recent studies have revealed the social aspects of bacterial life, such as biofilm formation. Biofilm formation is largely affected by the environment, and the mechanisms by which the gene expression of individual cells affects biofilm development have attracted interest. Environmental factors determine the cell's decision to form or leave a biofilm. In addition, the biofilm structure largely depends on the environment, implying that biofilms are shaped to adapt to local conditions. Second messengers such as cAMP and c-di-GMP are key factors that link environmental factors with gene regulation. Cell-to-cell communication is also an important factor in shaping the biofilm. In this short review, we will introduce the basics of biofilm formation and further discuss environmental factors that shape biofilm formation. Finally, the state-of-the-art tools that allow us investigate biofilms under various conditions are discussed.

  1. Holographic microrheology of biofilms

    Science.gov (United States)

    Chiong Cheong, Fook; Duarte, Simone; Grier, David

    2008-03-01

    We present microrheological measurements of polymeric matrices, including the extra-cellular polysaccharide gel synthesized by the dental pathogen S. mutans. As part of this study, we introduce the use of precision three-dimensional particle tracking based on video holographic microscopy. This technique offers nanometer-scale resolution at video rates, thereby providing detailed information on the gels' complex viscoelastic moduli, including insights into their heterogeneity. The particular application to dental biofilms complements previous studies based on macroscopic rheology, and demonstrates the utility of holographic microrheology for soft-matter physics and biomedical research.

  2. Experimental study of cadmium interaction with periphytic biofilms

    Energy Technology Data Exchange (ETDEWEB)

    Pokrovsky, O.S., E-mail: oleg@lmtg.obs-mip.fr [Geochimie et Biogeochimie Experimentale, UMR 5563, CNRS-OMP-Universite Toulouse, 14 Avenue Edouard Belin, 31400 Toulouse (France); Feurtet-Mazel, A. [Universite de Bordeaux 1, CNRS, UMR 5805 EPOC, Place du Dr Peyneau, 33120 Arcachon (France); Martinez, R.E. [Center for Applied Geosciences, Universitat Tuebingen, Sigwartstrasse 10, Tuebingen 72076 (Germany); Morin, S. [Unite de Recherche Reseaux, Epuration et Qualite des Eaux REQE, Cemagref, 50 Avenue de Verdun, F-33612 Cestas Cedex (France); Baudrimont, M. [Universite de Bordeaux 1, CNRS, UMR 5805 EPOC, Place du Dr Peyneau, 33120 Arcachon (France); Duong, T. [Universite de Bordeaux 1, CNRS, UMR 5805 EPOC, Place du Dr Peyneau, 33120 Arcachon (France)] [Institute of Environmental Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Cau Giay, Hanoi (Viet Nam); Coste, M. [Unite de Recherche Reseaux, Epuration et Qualite des Eaux REQE, Cemagref, 50 Avenue de Verdun, F-33612 Cestas Cedex (France)

    2010-03-15

    This study addresses the interaction of Cd with natural biofilms of periphytic diatoms grown during different seasons in metal-contaminated and metal-non-contaminated streams, along a tributary of the Lot River, France. Specifically, it aims to test whether the biofilms from contaminated sites have developed a protective mechanism due to high Cd exposure. Towards this goal, reversible adsorption experiments on untreated biofilms were performed in 0.01 M NaNO{sub 3} with a pH ranging from 2 to 8, Cd concentration from 0.5 to 10,000 {mu}g/L and exposure time from 1 to 24 h. Two types of experiments, pH-dependent adsorption edge and constant-pH 'Langmuirian'-type isotherms were conducted. Results were adequately modeled using a Linear Programming Model. It was found that the adsorption capacities of natural biofilm consortia with respect to Cd do not depend on season and are not directly linked to the growth environment. The biofilms grown in non-contaminated (4.6 ppb Cd in solid) and contaminated (570 ppb Cd in solid) settings exhibit similar adsorption capacities in the Cd concentration range in solution of 100-10,000 {mu}g/L but quite different capacities at low Cd concentration (0.5-100 {mu}g/L); unexpectedly, the non-contaminated biofilm adsorbs approximately 10 times more Cd than the contaminated one. It is therefore possible that the strong low-abundant ligands (for example, phosphoryl or sulfhydryls) are already metal-saturated on surfaces of biofilm grown in the contaminated site whereas these sites are still available for metal adsorption in samples grown in non-contaminated sites.

  3. Significance of biofilms in dentistry.

    Science.gov (United States)

    Wróblewska, Marta; Strużycka, Izabela; Mierzwińska-Nastalska, Elżbieta

    2015-01-01

    In the past decades significant scientific progress has taken place in the knowledge about biofilms. They constitute multilayer conglomerates of bacteria and fungi, surrounded by carbohydrates which they produce, as well as substances derived from saliva and gingival fluid. Modern techniques showed significant diversity of the biofilm environment and a system of microbial communication (quorum sensing), enhancing their survival. At present it is believed that the majority of infections, particularly chronic with exacerbations, are a result of biofilm formation, particularly in the presence of biomaterials. It should be emphasised that penetration of antibiotics and other antimicrobial agents into deeper layers of a biofilm is poor, causing therapeutic problems and necessitating sometimes removal of the implant or prosthesis. Biofilms play an increasing role in dentistry as a result of more and more broad use in dental practice of plastic and implantable materials. Biofilms are produced on the surfaces of teeth as dental plaque, in the para-nasal sinuses, on prostheses, dental implants, as well as in waterlines of a dental unit, constituting a particular risk for severely immunocompromised patients. New methods of therapy and prevention of infections linked to biofilms are under development.

  4. The MerR-like regulator BrlR confers biofilm tolerance by activating multidrug efflux pumps in Pseudomonas aeruginosa biofilms.

    Science.gov (United States)

    Liao, Julie; Schurr, Michael J; Sauer, Karin

    2013-08-01

    A defining characteristic of biofilms is antibiotic tolerance that can be up to 1,000-fold greater than that of planktonic cells. In Pseudomonas aeruginosa, biofilm tolerance to antimicrobial agents requires the biofilm-specific MerR-type transcriptional regulator BrlR. However, the mechanism by which BrlR mediates biofilm tolerance has not been elucidated. Genome-wide transcriptional profiling indicated that brlR was required for maximal expression of genes associated with antibiotic resistance, in particular those encoding the multidrug efflux pumps MexAB-OprM and MexEF-OprN. Chromatin immunoprecipitation (ChIP) analysis revealed a direct regulation of these genes by BrlR, with DNA binding assays confirming BrlR binding to the promoter regions of the mexAB-oprM and mexEF-oprN operons. Quantitative reverse transcriptase PCR (qRT-PCR) analysis further indicated BrlR to be an activator of mexAB-oprM and mexEF-oprN gene expression. Moreover, immunoblot analysis confirmed increased MexA abundance in cells overexpressing brlR. Inactivation of both efflux pumps rendered biofilms significantly more susceptible to five different classes of antibiotics by affecting MIC but not the recalcitrance of biofilms to killing by bactericidal agents. Overexpression of either efflux pump in a ΔbrlR strain partly restored tolerance of ΔbrlR biofilms to antibiotics. Expression of brlR in mutant biofilms lacking both efflux pumps partly restored antimicrobial tolerance of biofilms to wild-type levels. Our results indicate that BrlR acts as an activator of multidrug efflux pumps to confer tolerance to P. aeruginosa biofilms and to resist the action of antimicrobial agents.

  5. Two-dimensional and three-dimensional dynamic imaging of live biofilms in a microchannel by time-of-flight secondary ion mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Hua, Xin; Marshall, Matthew J.; Xiong, Yijia; Ma, Xiang; Zhou, Yufan; Tucker, Abigail E.; Zhu, Zihua; Liu, Songqin; Yu, Xiao-Ying

    2015-05-01

    A vacuum compatible microfluidic reactor, SALVI (System for Analysis at the Liquid Vacuum Interface) was employed for in situ chemical imaging of live biofilms using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Depth profiling by sputtering materials in sequential layers resulted in live biofilm spatial chemical mapping. 2D images were reconstructed to report the first 3D images of hydrated biofilm elucidating spatial and chemical heterogeneity. 2D image principal component analysis (PCA) was conducted among biofilms at different locations in the microchannel. Our approach directly visualized spatial and chemical heterogeneity within the living biofilm by dynamic liquid ToF-SIMS.

  6. Two-dimensional and three-dimensional dynamic imaging of live biofilms in a microchannel by time-of-flight secondary ion mass spectrometry.

    Science.gov (United States)

    Hua, Xin; Marshall, Matthew J; Xiong, Yijia; Ma, Xiang; Zhou, Yufan; Tucker, Abigail E; Zhu, Zihua; Liu, Songqin; Yu, Xiao-Ying

    2015-05-01

    A vacuum compatible microfluidic reactor, SALVI (System for Analysis at the Liquid Vacuum Interface), was employed for in situ chemical imaging of live biofilms using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Depth profiling by sputtering materials in sequential layers resulted in live biofilm spatial chemical mapping. Two-dimensional (2D) images were reconstructed to report the first three-dimensional images of hydrated biofilm elucidating spatial and chemical heterogeneity. 2D image principal component analysis was conducted among biofilms at different locations in the microchannel. Our approach directly visualized spatial and chemical heterogeneity within the living biofilm by dynamic liquid ToF-SIMS.

  7. Visualization of extracellular matrix components within sectioned Salmonella biofilms on the surface of human gallstones.

    Directory of Open Access Journals (Sweden)

    Joanna M Marshall

    Full Text Available Chronic carriage of Salmonella Typhi is mediated primarily through the formation of bacterial biofilms on the surface of cholesterol gallstones. Biofilms, by definition, involve the formation of a bacterial community encased within a protective macromolecular matrix. Previous work has demonstrated the composition of the biofilm matrix to be complex and highly variable in response to altered environmental conditions. Although known to play an important role in bacterial persistence in a variety of contexts, the Salmonella biofilm matrix remains largely uncharacterized under physiological conditions. Initial attempts to study matrix components and architecture of the biofilm matrix on gallstone surfaces were hindered by the auto-fluorescence of cholesterol. In this work we describe a method for sectioning and direct visualization of extracellular matrix components of the Salmonella biofilm on the surface of human cholesterol gallstones and provide a description of the major matrix components observed therein. Confocal micrographs revealed robust biofilm formation, characterized by abundant but highly heterogeneous expression of polysaccharides such as LPS, Vi and O-antigen capsule. CsgA was not observed in the biofilm matrix and flagellar expression was tightly restricted to the biofilm-cholesterol interface. Images also revealed the presence of preexisting Enterobacteriaceae encased within the structure of the gallstone. These results demonstrate the use and feasibility of this method while highlighting the importance of studying the native architecture of the gallstone biofilm. A better understanding of the contribution of individual matrix components to the overall biofilm structure will facilitate the development of more effective and specific methods to disrupt these bacterial communities.

  8. Novel microfluidic system for online monitoring of biofilm dynamics by electrical impedance spectroscopy and amperometry

    Science.gov (United States)

    Bruchmann, Julia; Sachsenheimer, Kai; Schwartz, Thomas; Rapp, Bastian E.

    2016-03-01

    Biofilm formation is ubiquitous in nature where microorganisms attach to surfaces and form highly adapted and protected communities. In technical and industrial systems like drinking water supply, food production or shipping industry biofilms are a major cause of product contamination, biofouling, and biocorrosion. Therefore, understanding of biofilm formation and means of preventing biofilm formation is important to develop novel biofilm treatment strategies. A system allowing directly online detection and monitoring biofilm formation is necessary. However, until today, there are little to none technical systems featuring a non-destructive real-time characterization of biofilm formation in a highthroughput manner. This paper presents such a microfluidic system based on electrical impedance spectroscopy (EIS) and amperomertic current measurement. The sensor consists of four modules, each housing 24 independent electrodes within 12 microfluidic channels. Attached biomass on the electrodes is monitored as increased inhibition in charge transfer by EIS and a change in metabolic activity is measured as change in produced electric current by amperometry. This modular sensor system is highly adaptable and suitable for a broad range of microbiological applications. Among others, biofilm formation processes can be characterized online, biofilm manipulation like inactivation or destabilization can be monitored in real-time and gene expression can be analyzed in parallel. The use of different electrode designs allows effective biofilm studies during all biofilm phases. The whole system was recently extended by an integrated pneumatic microfluidic pump which enables easy handling procedures. Further developments of this pumping module will allow a fully- automated computer-controlled valving and pumping.

  9. Fully printed flexible and disposable wireless cyclic voltammetry tag.

    Science.gov (United States)

    Jung, Younsu; Park, Hyejin; Park, Jin-Ah; Noh, Jinsoo; Choi, Yunchang; Jung, Minhoon; Jung, Kyunghwan; Pyo, Myungho; Chen, Kevin; Javey, Ali; Cho, Gyoujin

    2015-01-29

    A disposable cyclic voltammetry (CV) tag is printed on a plastic film by integrating wireless power transmitter, polarized triangle wave generator, electrochemical cell and signage through a scalable gravure printing method. By proximity of 13.56 MHz RF reader, the printed CV tag generates 320 mHz of triangular sweep wave from +500 mV to -500 mV which enable to scan a printed electrochemical cell in the CV tag. By simply dropping any specimen solution on the electrochemical cell in the CV tag, the presence of solutes in the solution can be detected and shown on the signage of the CV tag in five sec. 10 mM of N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD) was used as a standard solute to prove the working concept of fully printed disposable wireless CV tag. Within five seconds, we can wirelessly diagnose the presence of TMPD in the solution using the CV tag in the proximity of the 13.56 MHz RF reader. This fully printed and wirelessly operated flexible CV tag is the first of its kind and marks the path for the utilization of inexpensive and disposable wireless electrochemical sensor systems for initial diagnose hazardous chemicals and biological molecules to improve public hygiene and health.

  10. In vivo histamine voltammetry in the mouse premammillary nucleus.

    Science.gov (United States)

    Samaranayake, Srimal; Abdalla, Aya; Robke, Rhiannon; Wood, Kevin M; Zeqja, Anisa; Hashemi, Parastoo

    2015-06-07

    Histamine plays a major role in the mediation of allergic reactions such as peripheral inflammation. This classical monoamine is also a neurotransmitter involved in the central nervous system but its role in this context is poorly understood. Studying histamine neurotransmission is important due to its implications in many neurological disorders. The sensitivity, selectivity and high temporal resolution of fast scan cyclic voltammetry (FSCV) offer many advantages for studying electroactive neurotransmitters. Histamine has previously been studied with FSCV; however, the lack of a robust Faradaic electrochemical signal makes it difficult to selectively identify histamine in complex media, as found in vivo. In this work, we optimize an electrochemical waveform that provides a stimulation-locked and unique electrochemical signal towards histamine. We describe in vitro waveform optimization and a novel in vivo physiological model for stimulating histamine release in the mouse premammillary nucleus via stimulation of the medial forebrain bundle. We demonstrate that a robust signal can be used to effectively identify histamine and characterize its in vivo kinetics.

  11. Cyclic voltammetry and RBS study of paint components

    Energy Technology Data Exchange (ETDEWEB)

    Bowman, Lynn [Center for Irradiation of Materials, Alabama A and M University, 4900 Meridian Street, P.O. Box 1447, Normal, AL 35762 (United States); Spencer, Dirk [Plant and Soil Science Department, Alabama A and M University, Normal, AL 35762-1447 (United States); Muntele, Claudiu [Center for Irradiation of Materials, Alabama A and M University, 4900 Meridian Street, P.O. Box 1447, Normal, AL 35762 (United States)]. E-mail: claudiu@cim.aamu.edu; Muntele, Iulia [Center for Irradiation of Materials, Alabama A and M University, 4900 Meridian Street, P.O. Box 1447, Normal, AL 35762 (United States); Ila, D. [Center for Irradiation of Materials, Alabama A and M University, 4900 Meridian Street, P.O. Box 1447, Normal, AL 35762 (United States)

    2007-08-15

    Heavy metals and metalloid ions are found in environmental matrices. The most toxic are lead, cadmium and mercury. These three heavy metals have no biological function and are toxic at all concentrations. Lead is one of the most insidious heavy metals and is introduced into the environment by many different means. It persists in both urban and rural settings, being found in paint chips, pottery, crystal and pharmaceutical and nutritional products. The analysis of heavy elements such as lead in soil is of particular importance [W.T. Sturges, R.M. Harrison, Sci. Total Environ. 44 (3) (1985) 225; M.L. Lepow, L. Bruckman, M. Gillette, S. Markowitz, R. Robino, J. Kapish, Environ. Res. 10 (3) (1975) 415; A.E. Daniels, J.R. Kominsky, P.J. Clark, J. Hazard. Mater. B 87 (2001) 117; G. Hutter, D. Moshman, J. Hazard. Mater. 40 (1995) 1]. In preparing the methods for lead detection in paint, we have used Rutherford backscattering spectrometry (RBS) in order to study the type and amount of heavy metal content in paint samples collected at various sites in the historic campus at A and M University (AAMU). We will show the results of our study with emphasis on comparison of what we learned about presence of lead in paints using our ion beam methods compared with the analysis of lead in paints using cyclic voltammetry.

  12. Biofilm models for the practitioner

    DEFF Research Database (Denmark)

    Morgenroth, Eberhard Friedrich; van Loosdrecht, M. C. M.; Wanner, O.

    2000-01-01

    Even though mathematical biofilm models are extensively used in biofilm research, there has been very little application of these models in the engineering practice so far. However, practitioners would be interested in models that can be used as tools to control plant operation under dynamic...... conditions or to help them handle complex interactions between particle removal, carbon oxidation, nitrification, denitrification and biological phosphorus removal. But even though there is a whole range of biofilm models available, it is difficult for the practitioner to select the appropriate modeling...

  13. Antibiotic resistance of bacterial biofilms

    DEFF Research Database (Denmark)

    Hoiby, N.; Bjarnsholt, T.; Givskov, M.

    2010-01-01

    and other components of the body's defence system. The persistence of, for example, staphylococcal infections related to foreign bodies is due to biofilm formation. Likewise, chronic Pseudomonas aeruginosa lung infection in cystic fibrosis patients is caused by biofilm-growing mucoid strains...... to antibiotics. Biofilm growth is associated with an increased level of mutations as well as with quorum-sensing-regulated mechanisms. Conventional resistance mechanisms such as chromosomal beta-lactamase, upregulated efflux pumps and mutations in antibiotic target molecules in bacteria also contribute...

  14. CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein, CsrA.

    Science.gov (United States)

    Willias, Stephan P; Chauhan, Sadhana; Lo, Chien-Chi; Chain, Patrick S G; Motin, Vladimir L

    2015-01-01

    The natural transmission of Yersinia pestis is reliant upon biofilm blockage of the flea vector. However, the environmentally-responsive adaptive regulators which facilitate Y. pestis biofilm production in accordance with the flea midgut milieu are not well understood. We seek to establish the impact of available carbon source metabolism and storage upon Y. pestis biofilm production. Our findings demonstrate that Y. pestis biofilm production is subject to carbon catabolite regulation in which the presence of glucose impairs biofilm production; whereas, the sole metabolism of alternate carbon sources promotes robust biofilm formation. This observation is facilitated by the cAMP receptor protein, CRP. In accordance with a stark growth defect, deletion of crp in both CO92 and KIM6+ Y. pestis strains significantly impaired biofilm production when solely utilizing alternate carbon sources. Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production. Furthermore, CRP did not alter mRNA abundance of previously-characterized hms biofilm synthesis and regulation factors. Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile. Additionally, we assessed the impact of the carbon storage regulator protein, CsrA, upon Y. pestis biofilm production. Contrary to what has been described for E. coli, Y. pestis biofilm formation was found to be enhanced by CsrA. Regardless of media composition and available carbon source, deletion of csrA significantly impaired Y. pestis biofilm production. CsrA was found to promote Y. pestis biofilm production independent of glycogen regulation. Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance. However, deletion of hmsP in the csrA-deficient mutant enabled excessive biofilm production, suggesting Csr

  15. Anti-Biofilm Performance of Three Natural Products against Initial Bacterial Attachment

    Directory of Open Access Journals (Sweden)

    Keith R. Stokes

    2013-11-01

    Full Text Available Marine bacteria contribute significantly towards the fouling consortium, both directly (modern foul release coatings fail to prevent “slime” attachment and indirectly (biofilms often excrete chemical cues that attract macrofouling settlement. This study assessed the natural product anti-biofilm performance of an extract of the seaweed, Chondrus crispus, and two isolated compounds from terrestrial sources, (+-usnic acid and juglone, against two marine biofilm forming bacteria, Cobetia marina and Marinobacter hydrocarbonoclasticus. Bioassays were developed using quantitative imaging and fluorescent labelling to test the natural products over a range of concentrations against initial bacterial attachment. All natural products affected bacterial attachment; however, juglone demonstrated the best anti-biofilm performance against both bacterial species at a concentration range between 5–20 ppm. In addition, for the first time, a dose-dependent inhibition (hormetic response was observed for natural products against marine biofilm forming bacteria.

  16. Morphological evidence of biofilm formation in Greenlanders with chronic suppurative otitis media

    DEFF Research Database (Denmark)

    Homøe, Preben; Bjarnsholt, Thomas; Wessman, Marcus;

    2009-01-01

    Biofilm may explain the recurrences and recalcitrant episodes of otorrhea in chronic suppurative otitis media (CSOM). This study investigates bacterial biofilm in Greenlanders with CSOM and chronic otitis media with effusion (COME). The study is partly blinded, prospective and retrospective. Six...... were analyzed with microscopy and peptide nucleic acid fluorescence in situ hybridization (PNA-FISH). Biofilm was confirmed in 83% of CSOM smears but in none of the COME smears. Mucosal biofilm was confirmed in 80% of the biopsies from adults with CSOM. This study provides direct morphological evidence...... of biofilm in samples from human CSOM. This may help to explain the microbiological mechanisms of the disease and alter the treatment strategy in the future....

  17. Comparison of the In vitro Activity of Five Antimicrobial Drugs against Staphylococcus pseudintermedius and Staphylococcus aureus Biofilms

    Science.gov (United States)

    Ferran, Aude A.; Liu, JingJing; Toutain, Pierre-Louis; Bousquet-Mélou, Alain

    2016-01-01

    Resistance in canine pathogenic staphylococci is necessitating re-evaluation of the current antimicrobial treatments especially for biofilm-associated infections. Long, repeated treatments are often required to control such infections due to the tolerance of bacteria within the biofilm. To comply with the goal of better antibiotic stewardship in veterinary medicine, the efficacies of the available drugs need to be directly assessed on bacterial biofilms. We compared the activities of amoxicillin, cefalexin, clindamycin, doxycycline, and marbofloxacin on in vitro biofilms of Staphylococcus pseudintermedius and Staphylococcus aureus. Exposure of biofilms for 15 h to maximum concentrations of the antibiotics achievable in canine plasma only reduced biofilm bacteria by 0.5–2.0 log10 CFU, compared to the control, except for marbofloxacin which reduced S. aureus biofilms by 5.4 log10 CFU. Two-antibiotic combinations did not improve, and even decreased, bacterial killing. In comparison, 5 min-exposure to 2% chlorhexidine reduced biofilms of the two tested strains by 4 log10 CFU. Our results showed that S. pseudintermedius and S. aureus biofilms were highly tolerant to all the drugs tested, consistent with the treatment failures observed in practice. Under our in vitro conditions, the use of chlorhexidine was more efficacious than antimicrobials to reduce S. pseudintermedius biofilm. PMID:27531995

  18. A quantitative method to measure biofilm removal efficiency from complex biomaterial surfaces using SEM and image analysis

    Science.gov (United States)

    Vyas, N.; Sammons, R. L.; Addison, O.; Dehghani, H.; Walmsley, A. D.

    2016-01-01

    Biofilm accumulation on biomaterial surfaces is a major health concern and significant research efforts are directed towards producing biofilm resistant surfaces and developing biofilm removal techniques. To accurately evaluate biofilm growth and disruption on surfaces, accurate methods which give quantitative information on biofilm area are needed, as current methods are indirect and inaccurate. We demonstrate the use of machine learning algorithms to segment biofilm from scanning electron microscopy images. A case study showing disruption of biofilm from rough dental implant surfaces using cavitation bubbles from an ultrasonic scaler is used to validate the imaging and analysis protocol developed. Streptococcus mutans biofilm was disrupted from sandblasted, acid etched (SLA) Ti discs and polished Ti discs. Significant biofilm removal occurred due to cavitation from ultrasonic scaling (p SLA surface images were 0.80 ± 0.18 and 0.62 ± 0.20 respectively and 0.74 ± 0.13 and 0.86 ± 0.09 respectively for polished surfaces. Cavitation has potential to be used as a novel way to clean dental implants. This imaging and analysis method will be of value to other researchers and manufacturers wishing to study biofilm growth and removal. PMID:27601281

  19. Comparison of the In vitro Activity of Five Antimicrobial Drugs against Staphylococcus pseudintermedius and Staphylococcus aureus Biofilms.

    Science.gov (United States)

    Ferran, Aude A; Liu, JingJing; Toutain, Pierre-Louis; Bousquet-Mélou, Alain

    2016-01-01

    Resistance in canine pathogenic staphylococci is necessitating re-evaluation of the current antimicrobial treatments especially for biofilm-associated infections. Long, repeated treatments are often required to control such infections due to the tolerance of bacteria within the biofilm. To comply with the goal of better antibiotic stewardship in veterinary medicine, the efficacies of the available drugs need to be directly assessed on bacterial biofilms. We compared the activities of amoxicillin, cefalexin, clindamycin, doxycycline, and marbofloxacin on in vitro biofilms of Staphylococcus pseudintermedius and Staphylococcus aureus. Exposure of biofilms for 15 h to maximum concentrations of the antibiotics achievable in canine plasma only reduced biofilm bacteria by 0.5-2.0 log10 CFU, compared to the control, except for marbofloxacin which reduced S. aureus biofilms by 5.4 log10 CFU. Two-antibiotic combinations did not improve, and even decreased, bacterial killing. In comparison, 5 min-exposure to 2% chlorhexidine reduced biofilms of the two tested strains by 4 log10 CFU. Our results showed that S. pseudintermedius and S. aureus biofilms were highly tolerant to all the drugs tested, consistent with the treatment failures observed in practice. Under our in vitro conditions, the use of chlorhexidine was more efficacious than antimicrobials to reduce S. pseudintermedius biofilm.

  20. Biofilm control with natural and genetically-modified phages.

    Science.gov (United States)

    Motlagh, Amir Mohaghegh; Bhattacharjee, Ananda Shankar; Goel, Ramesh

    2016-04-01

    Bacteriophages, as the most dominant and diverse entities in the universe, have the potential to be one of the most promising therapeutic agents. The emergence of multidrug-resistant bacteria and the antibiotic crisis in the last few decades have resulted in a renewed interest in phage therapy. Furthermore, bacteriophages, with the capacity to rapidly infect and overcome bacterial resistance, have demonstrated a sustainable approach against bacterial pathogens-particularly in biofilm. Biofilm, as complex microbial communities located at interphases embedded in a matrix of bacterial extracellular polysaccharide substances (EPS), is involved in health issues such as infections associated with the use of biomaterials and chronic infections by multidrug resistant bacteria, as well as industrial issues such as biofilm formation on stainless steel surfaces in food industry and membrane biofouling in water and wastewater treatment processes. In this paper, the most recent studies on the potential of phage therapy using natural and genetically-modified lytic phages and their associated enzymes in fighting biofilm development in various fields including engineering, industry, and medical applications are reviewed. Phage-mediated prevention approaches as an indirect phage therapy strategy are also explored in this review. In addition, the limitations of these approaches and suggestions to overcome these constraints are discussed to enhance the efficiency of phage therapy process. Finally, future perspectives and directions for further research towards a better understanding of phage therapy to control biofilm are recommended.

  1. Regulation of biofilm formation in Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Simm, Roger; Ahmad, Irfan; Rhen, Mikael; Le Guyon, Soazig; Römling, Ute

    2014-01-01

    In animals, plants and the environment, Salmonella enterica serovar Typhimurium forms the red dry and rough (rdar) biofilm characterized by extracellular matrix components curli and cellulose. With complex expression control by at least ten transcription factors, the bistably expressed orphan response regulator CsgD directs rdar morphotype development. CsgD expression is an integral part of the Hfq regulon and the complex cyclic diguanosine monophosphate signaling network partially controlled by the global RNA-binding protein CsrA. Cell wall turnover and the periplasmic redox status regulate csgD expression on a post-transcriptional level by unknown mechanisms. Furthermore, phosphorylation of CsgD is a potential inactivation and degradation signal in biofilm dissolution. Including complex incoherent feed-forward loops, regulation of biofilm formation versus motility and virulence is of recognized complexity.

  2. Probing electric fields inside microfluidic channels during electroosmotic flow with fast-scan cyclic voltammetry.

    Science.gov (United States)

    Forry, Samuel P; Murray, Jacqueline R; Heien, Michael L A V; Locascio, Laurie E; Wightman, R Mark

    2004-09-01

    Fast-scan cyclic voltammetry (FSCV) at carbon-fiber microelectrodes was used in microfluidic channels. This method offers the advantage that it can resolve electroactive species not separated in the channel. In addition, this method provides a route to investigate the distribution of applied electrophoretic fields in microfluidic channels. To probe this, microelectrodes were inserted at various distances into channels and cyclic voltammograms recorded at 300 V/s were repeated at 0.1-s intervals. The use of a battery-powered laptop computer and potentiostat provided galvanic isolation between the applied electrophoretic field and the electrochemical measurements. In the absence of an external field, the peak potential for oxidation of the test solute, Ru(bpy)3(2+), was virtually unaltered by insertion of the microelectrode tip into the channel. When an electrophoretic field was applied, the peak potential for Ru(bpy)3(2+) oxidation shifted to more positive potentials in a manner that was directly proportional to the field in the channel. The shifts in peak potential observed with FSCV enabled direct compensation of the applied electrochemical potential. This approach was used to explore the electrophoretic field at the channel terminus. It was found to persist for more than 50 microm from the channel terminus. In addition, the degree of analyte dispersion was found to depend critically on the electrode position outside the channel.

  3. Metabolism links bacterial biofilms and colon carcinogenesis.

    Science.gov (United States)

    Johnson, Caroline H; Dejea, Christine M; Edler, David; Hoang, Linh T; Santidrian, Antonio F; Felding, Brunhilde H; Ivanisevic, Julijana; Cho, Kevin; Wick, Elizabeth C; Hechenbleikner, Elizabeth M; Uritboonthai, Winnie; Goetz, Laura; Casero, Robert A; Pardoll, Drew M; White, James R; Patti, Gary J; Sears, Cynthia L; Siuzdak, Gary

    2015-06-02

    Bacterial biofilms in the colon alter the host tissue microenvironment. A role for biofilms in colon cancer metabolism has been suggested but to date has not been evaluated. Using metabolomics, we investigated the metabolic influence that microbial biofilms have on colon tissues and the related occurrence of cancer. Patient-matched colon cancers and histologically normal tissues, with or without biofilms, were examined. We show the upregulation of polyamine metabolites in tissues from cancer hosts with significant enhancement of N(1), N(12)-diacetylspermine in both biofilm-positive cancer and normal tissues. Antibiotic treatment, which cleared biofilms, decreased N(1), N(12)-diacetylspermine levels to those seen in biofilm-negative tissues, indicating that host cancer and bacterial biofilm structures contribute to the polyamine metabolite pool. These results show that colonic mucosal biofilms alter the cancer metabolome to produce a regulator of cellular proliferation and colon cancer growth potentially affecting cancer development and progression.

  4. Metabolism links bacterial biofilms and colon carcinogenesis

    Science.gov (United States)

    Johnson, Caroline H.; Dejea, Christine M.; Edler, David; Hoang, Linh T.; Santidrian, Antonio F.; Felding, Brunhilde H.; Cho, Kevin; Wick, Elizabeth C.; Hechenbleikner, Elizabeth M.; Uritboonthai, Winnie; Goetz, Laura; Casero, Robert A.; Pardoll, Drew M.; White, James R.; Patti, Gary J.; Sears, Cynthia L.; Siuzdak, Gary

    2015-01-01

    SUMMARY Bacterial biofilms in the colon alter the host tissue microenvironment. A role for biofilms in colon cancer metabolism has been suggested but to date has not been evaluated. Using metabolomics, we investigated the metabolic influence that microbial biofilms have on colon tissues and the related occurrence of cancer. Patient-matched colon cancers and histologically normal tissues, with or without biofilms, were examined. We show the upregulation of polyamine metabolites in tissues from cancer hosts with significant enhancement of N1, N12-diacetylspermine in both biofilm positive cancer and normal tissues. Antibiotic treatment, which cleared biofilms, decreased N1, N12-diacetylspermine levels to those seen in biofilm negative tissues, indicating that host cancer and bacterial biofilm structures contribute to the polyamine metabolite pool. These results show that colonic mucosal biofilms alter the cancer metabolome, to produce a regulator of cellular proliferation and colon cancer growth potentially affecting cancer development and progression. PMID:25959674

  5. The ``Swiss cheese'' instability of bacterial biofilms

    Science.gov (United States)

    Jang, Hongchul; Rusconi, Roberto; Stocker, Roman

    2012-11-01

    Bacteria often adhere to surfaces, where they develop polymer-encased communities (biofilms) that display dramatic resistance to antibiotic treatment. A better understanding of cell detachment from biofilms may lead to novel strategies for biofilm disruption. Here we describe a new detachment mode, whereby a biofilm develops a nearly regular array of ~50-100 μm holes. Using surface-treated microfluidic devices, we create biofilms of controlled shape and size. After the passage of an air plug, the break-up of the residual thin liquid film scrapes and rearranges bacteria on the surface, such that a ``Swiss cheese'' pattern is left in the residual biofilm. Fluorescent staining of the polymeric matrix (EPS) reveals that resistance to cell dislodgement correlates with local biofilm age, early settlers having had more time to hunker down. Because few survivors suffice to regrow a biofilm, these results point at the importance of considering microscale heterogeneity in assessing the effectiveness of biofilm removal strategies.

  6. Effect of calcium on moving-bed biofilm reactor biofilms.

    Science.gov (United States)

    Goode, C; Allen, D G

    2011-03-01

    The effect of calcium concentration on the biofilm structure, microbiology, and treatment performance was evaluated in a moving-bed biofilm reactor. Three experiments were conducted in replicate laboratory-scale reactors to determine if wastewater calcium is an important variable for the design and optimization of these reactors. Biofilm structural properties, such as thickness, oxygen microprofiles, and the composition of extracellular polymeric substances (EPS) were affected by increasing calcium concentrations. Above a threshold concentration of calcium between 1 and 50 mg/L, biofilms became thicker and denser, with a shift toward increasingly proteinaceous EPS at higher calcium concentrations up to 200 mgCa2+/L. At 300 mgCa2+/L, biofilms were found to become primarily composed of inorganic calcium precipitates. Microbiology was assessed through microscopy, denaturing grade gel electrophoresis, and enumeration of higher organisms. Higher calcium concentrations were found to change the bacterial community and promote the abundant growth of filamentous organisms and various protazoa and metazoan populations. The chemical oxygen demand removal efficiency was improved for reactors at calcium concentrations of 50 mg/L and above. Reactor effluents for the lowest calcium concentration (1 mgCa2+/L) were found to be turbid (>50 NTU), as a result of the detachment of small and poorly settling planktonic biomass, whereas higher concentrations promoted settling of the suspended phase. In general, calcium was found to be an important variable causing significant changes in biofilm structure and reactor function.

  7. Biofilm removal technique using sands as a research tool for accessing microbial attachment on surface

    Directory of Open Access Journals (Sweden)

    Nathanon Trachoo

    2004-01-01

    Full Text Available Biofilms have profound impacts on improved survival of the constituent microorganisms in nature. Biofilms were believed to protect constituent microorganisms from sanitizer treatment, provide a more suitable habitat for microorganisms, and become a site for genetic material exchanges between microorganisms. As we realize more about the significance of biofilm, methods used for biofilm study should be consistently developed and evaluated. To determine microbial attachment on surfaces, usually biofilms are grown on substratum surfaces and removed by vortexing with glass beads or scraping. However, scraping is not as effective as vortexing with glass beads. Another approach is direct-agar overlaying which cannot be used with high density biofilm. In this experiment, we compared effectiveness of glass beads (298±28 μm in diameter and sands (width: 221±55 μm and length: 329±118 μm in removing biofilm of Pseudomonas aeruginosa by vortexing method. The results suggested that acid-washed sands, which are significantly less inexpensive than glass beads, were as effective as (P>0.05 analytical grade glass beads in Pseudomonas aeruginosa biofilm removal without inhibiting growth of the organism.

  8. Monoclonal antibodies against DNA-binding tips of DNABII proteins disrupt biofilms in vitro and induce bacterial clearance in vivo

    Directory of Open Access Journals (Sweden)

    Laura A. Novotny

    2016-08-01

    Full Text Available The vast majority of chronic and recurrent bacterial diseases are attributed to the presence of a recalcitrant biofilm that contributes significantly to pathogenesis. As such, these diseases will require an innovative therapeutic approach. We targeted DNABII proteins, an integral component of extracellular DNA (eDNA which is universally found as part of the pathogenic biofilm matrix to develop a biofilm disrupting therapeutic. We show that a cocktail of monoclonal antibodies directed against specific epitopes of a DNABII protein is highly effective to disrupt diverse biofilms in vitro as well as resolve experimental infection in vivo, in both a chinchilla and murine model. Combining this monoclonal antibody cocktail with a traditional antibiotic to kill bacteria newly released from the biofilm due to the action of the antibody cocktail was highly effective. Our results strongly support these monoclonal antibodies as attractive candidates for lead optimization as a therapeutic for resolution of bacterial biofilm diseases.

  9. Manganese Detection with a Metal Catalyst Free Carbon Nanotube Electrode: Anodic versus Cathodic Stripping Voltammetry.

    Science.gov (United States)

    Yue, Wei; Bange, Adam; Riehl, Bill L; Riehl, Bonnie D; Johnson, Jay M; Papautsky, Ian; Heineman, William R

    2012-10-01

    Anodic stripping voltammetry (ASV) and cathodic stripping voltammetry (CSV) were used to determine Mn concentration using metal catalyst free carbon nanotube (MCFCNT) electrodes and square wave stripping voltammetry (SWSV). The MCFCNTs are synthesized using a Carbo Thermal Carbide Conversion method which results in a material that does not contain residual transition metals. Detection limits of 120 nM and 93 nM were achieved for ASV and CSV, respectively, with a deposition time of 60 s. CSV was found to be better than ASV in Mn detection in many aspects, such as limit of detection and sensitivity. The CSV method was used in pond water matrix addition measurements.

  10. Manganese Detection with a Metal Catalyst Free Carbon Nanotube Electrode: Anodic versus Cathodic Stripping Voltammetry

    Science.gov (United States)

    Yue, Wei; Bange, Adam; Riehl, Bill L.; Riehl, Bonnie D.; Johnson, Jay M.; Papautsky, Ian; Heineman, William R.

    2013-01-01

    Anodic stripping voltammetry (ASV) and cathodic stripping voltammetry (CSV) were used to determine Mn concentration using metal catalyst free carbon nanotube (MCFCNT) electrodes and square wave stripping voltammetry (SWSV). The MCFCNTs are synthesized using a Carbo Thermal Carbide Conversion method which results in a material that does not contain residual transition metals. Detection limits of 120 nM and 93 nM were achieved for ASV and CSV, respectively, with a deposition time of 60 s. CSV was found to be better than ASV in Mn detection in many aspects, such as limit of detection and sensitivity. The CSV method was used in pond water matrix addition measurements. PMID:24235806

  11. Rapid depletion of dissolved oxygen in 96 well microtitre plate Staphylococcus epidermidis biofilm assays promotes biofilm development and is influenced by inoculum cell concentration

    OpenAIRE

    Cotter, John J.; O'Gara, James P.; Casey, Eoin

    2009-01-01

    Biofilm-related research using 96-well microtiter plates involves static incubation of plates indiscriminate of environmental conditions, making oxygen availability an important variable which has not been considered to date. By directly measuring dissolved oxygen concentration over time we report here that dissolved oxygen is rapidly consumed in Staphylococcus epidermidis biofilm cultures grown in 96-well plates irrespective of the oxygen concentration in the gaseous environment in which the...

  12. Differential growth of wrinkled biofilms

    CERN Document Server

    Espeso, D R; Einarsson, B

    2015-01-01

    Biofilms are antibiotic-resistant bacterial aggregates that grow on moist surfaces and can trigger hospital-acquired infections. They provide a classical example in biology where the dynamics of cellular communities may be observed and studied. Gene expression regulates cell division and differentiation, which affect the biofilm architecture. Mechanical and chemical processes shape the resulting structure. We gain insight into the interplay between cellular and mechanical processes during biofilm development on air-agar interfaces by means of a hybrid model. Cellular behavior is governed by stochastic rules informed by a cascade of concentration fields for nutrients, waste and autoinducers. Cellular differentiation and death alter the structure and the mechanical properties of the biofilm, which is deformed according to Foppl-Von Karman equations informed by cellular processes and the interaction with the substratum. Stiffness gradients due to growth and swelling produce wrinkle branching. We are able to repr...

  13. Nanotechnology: Role in dental biofilms

    Directory of Open Access Journals (Sweden)

    Bhardwaj Sonia

    2009-01-01

    Full Text Available Biofilms are surface- adherent populations of microorganisms consisting of cells, water and extracellular matrix material Nanotechnology is promising field of science which can guide our understanding of the role of interspecies interaction in the development of biofilm. Streptococcus mutans with other species of bacteria has been known to form dental biofilm. The correlation between genetically modified bacteria Streptococcus mutans and nanoscale morphology has been assessed using AFMi.e atomic force microscopy. Nanotechnology application includes 16 O/ 18 O reverse proteolytic labeling,use of quantum dots for labeling of bacterial cells, selective removal of cariogenic bacteria while preserving the normal oral flora and silver antimicrobial nanotechnology against pathogens associated with biofilms. The future comprises a mouthwash full of smart nanomachines which can allow the harmless flora of mouth to flourish in a healthy ecosystem

  14. Synergistic Interactions in Multispecies Biofilms

    DEFF Research Database (Denmark)

    Ren, Dawei

    between plasmid host range and composition of the recipient community was investigated in Manuscript 5 by comparing plasmid permissiveness in single populations and in a microbial community composed of 15 soil strains. By use of flow cytometry (FCM) and 16S rRNA gene sequencing, the IncP1 plasmid, pKJK10...... bacterial species, the study to elucidate the impact of interaction networks on the multispecies biofilms in natural ecosystems, especially in soil, is still at an early stage. The diverse patterns of interactions within the mixed communities as well as the predatorprey relationship between protozoa...... interactions in this four-species biofilm model community. Manuscript 2 presents the further application of this developed approach on evaluating the synergistic/antagonistic interactions in multispecies biofilms composed of seven soil isolates. 63% of the four-species biofilms were found to interact...

  15. Hydrodynamics of catheter biofilm formation

    CERN Document Server

    Sotolongo-Costa, Oscar; Rodriguez-Perez, Daniel; Martinez-Escobar, Sergio; Fernandez-Barbero, Antonio

    2009-01-01

    A hydrodynamic model is proposed to describe one of the most critical problems in intensive medical care units: the formation of biofilms inside central venous catheters. The incorporation of approximate solutions for the flow-limited diffusion equation leads to the conclusion that biofilms grow on the internal catheter wall due to the counter-stream diffusion of blood through a very thin layer close to the wall. This biological deposition is the first necessary step for the subsequent bacteria colonization.

  16. Critical review on biofilm methods

    DEFF Research Database (Denmark)

    Azeredo, Joana; F. Azevedo, Nuno; Briandet, Romain

    2017-01-01

    Biofilms are widespread in nature and constitute an important strategy implemented by microorganisms to survive in sometimes harsh environmental conditions. They can be beneficial or have a negative impact particularly when formed in industrial settings or on medical devices. As such, research in...... and limitations of several methods. Accordingly, this review aims at helping scientists in finding the most appropriate and up-to-date methods to study their biofilms....

  17. Biofilm-specific antibiotic tolerance and resistance.

    Science.gov (United States)

    Olsen, I

    2015-05-01

    Biofilms are heterogeneous structures composed of bacterial cells surrounded by a matrix and attached to solid surfaces. The bacteria here are 100 to 1,000 times more tolerant to antimicrobials than corresponding planktonic cells. Biofilms can be difficult to eradicate when they cause biofilm-related diseases, e.g., implant infections, cystic fibrosis, urinary tract infections, and periodontal diseases. A number of phenotypic features of the biofilm can be involved in biofilm-specific tolerance and resistance. Little is known about the molecular mechanisms involved. The current review deals with both phenotypic and molecular mechanisms of biofilm-specific antibiotic tolerance and resistance.

  18. Effect of chemical sanitizers with and without ultrasonication on Listeria monocytogenes as a biofilm within polyvinyl chloride drain pipes

    Science.gov (United States)

    As part of a biofilm in a floor drain, L. monocytogenes is exceedingly difficult to eradicate with standard sanitizing protocols. The objective of these studies was to test the use of ultra-sonication to break up biofilm architecture allowing chemical sanitizers to contact cells directly. L. monoc...

  19. Theory of linear sweep voltammetry with diffuse charge: unsupported electrolytes, thin films, and leaky membranes

    CERN Document Server

    Yan, David; Pugh, Mary C; Dawson, Francis P

    2016-01-01

    Linear sweep and cyclic voltammetry techniques are important tools for electrochemists and have a variety of applications in engineering. Voltammetry has classically been treated with the Randles-Sevcik equation, which assumes an electroneutral supported electrolyte. No general theory of linear-sweep voltammetry is available, however, for unsupported electrolytes and for other situations where diffuse charge effects play a role. In this paper, we provide a historical review of previous models and experiments and present a comprehensive mathematical theory of voltammetry in electrochemical cells with diffuse charge. We solve the time-dependent Poisson-Nernst-Planck (PNP) equations with generalized Frumkin-Butler-Volmer (FBV) boundary conditions, and show theoretical and simulated current-voltage curves for liquid and solid thin films, cells with blocking electrodes, and membranes with fixed background charge. The full range of dimensionless parameters is considered, including the dimensionless Debye screening ...

  20. Electrolyte Effects on the Cyclic Voltammetry of TCNQ (Tetracyanoquinodimethane) and TCNE (Tetracyanoethylene).

    Science.gov (United States)

    1986-07-30

    to form two radical anions. In this note we report a study by cyclic voltammetry of the reductions of TCNQ and TCNE to their radical anions and dianions at platinum and glassy carbon electrodes in acetonitrile.

  1. Strategies for combating bacterial biofilm infections

    Institute of Scientific and Technical Information of China (English)

    Hong Wu; Claus Moser; Heng-Zhuang Wang; Niels Hiby; Zhi-Jun Song

    2015-01-01

    Formation of biofilm is a survival strategy for bacteria and fungi to adapt to their living environment, especially in the hostile environment. Under the protection of biofilm, microbial cells in biofilm become tolerant and resistant to antibiotics and the immune responses, which increases the difficulties for the clinical treatment of biofilm infections. Clinical and laboratory investigations demonstrated a perspicuous correlation between biofilm infection and medical foreign bodies or indwelling devices. Clinical observations and experimental studies indicated clearly that antibiotic treatment alone is in most cases insufficient to eradicate biofilm infections. Therefore, to effectively treat biofilm infections with currently available antibiotics and evaluate the outcomes become important and urgent for clinicians. The review summarizes the latest progress in treatment of clinical biofilm infections and scientific investigations, discusses the diagnosis and treatment of different biofilm infections and introduces the promising laboratory progress, which may contribute to prevention or cure of biofilm infections. We conclude that, an efficient treatment of biofilm infections needs a well-established multidisciplinary collaboration, which includes removal of the infected foreign bodies, selection of biofilm-active, sensitive and well-penetrating antibiotics, systemic or topical antibiotic administration in high dosage and combinations, and administration of anti-quorum sensing or biofilm dispersal agents.

  2. The impact of shearing flows on electroactive biofilm formation, structure, and current generation

    Science.gov (United States)

    Jones, A.-Andrew; Buie, Cullen

    2016-11-01

    A special class of bacteria exist that directly produce electricity. First explored in 1911, these electroactive bacteria catalyze hydrocarbons and transport electrons directly to a metallic electron acceptor forming thicker biofilms than other species. Electroactive bacteria biofilms are thicker because they are not limited by transport of oxygen or other terminal electron acceptors. Electroactive bacteria can produce power in fuel cells. Power production is limited in fuel cells by the bacteria's inability to eliminate protons near the insoluble electron acceptor not utilized in the wild. To date, they have not been successfully evolved or engineered to overcome this limit. This limitation may be overcome by enhancing convective mass transport while maintaining substantial biomass within the biofilm. Increasing convective mass transport increases shear stress. A biofilm may respond to increased shear by changing biomass, matrix, or current production. In this study, a rotating disk electrode is used to separate nutrient from physical stress. This phenomenon is investigated using the model electroactive bacterium Geobacter sulfurreducens at nutrient loads comparable to flow-through microbial fuel cells. We determine biofilm structure experimentally by measuring the porosity and calculating the tortuosity from confocal microscope images. Biofilm adaptation for electron transport is quantified using electrical impedance spectroscopy. Our ultimate objective is a framework relating biofilm thickness, porosity, shear stress and current generation for the optimization of bioelectrochemical systems The Alfred P Sloan Foundation MPHD Program.

  3. Evaluation of microbial biofilm communities from an Alberta oil sands tailings pond.

    Science.gov (United States)

    Golby, Susanne; Ceri, Howard; Gieg, Lisa M; Chatterjee, Indranil; Marques, Lyriam L R; Turner, Raymond J

    2012-01-01

    Bitumen extraction from the oil sands of Alberta has resulted in millions of cubic meters of waste stored on-site in tailings ponds. Unique microbial ecology is expected in these ponds, which may be key to their bioremediation potential. We considered that direct culturing of microbes from a tailings sample as biofilms could lead to the recovery of microbial communities that provide good representation of the ecology of the tailings. Culturing of mixed species biofilms in vitro using the Calgary Biofilm Device (CBD) under aerobic, microaerobic, and anaerobic growth conditions was successful both with and without the addition of various growth nutrients. Denaturant gradient gel electrophoresis and 16S rRNA gene pyrotag sequencing revealed that unique mixed biofilm communities were recovered under each incubation condition, with the dominant species belonging to Pseudomonas, Thauera, Hydrogenophaga, Rhodoferax, and Acidovorax. This work used an approach that allowed organisms to grow as a biofilm directly from a sample collected of their environment, and the biofilms cultivated in vitro were representative of the endogenous environmental community. For the first time, representative environmental mixed species biofilms have been isolated and grown under laboratory conditions from an oil sands tailings pond environment and a description of their composition is provided.

  4. Anthranilate deteriorates the structure of Pseudomonas aeruginosa biofilms and antagonizes the biofilm-enhancing indole effect.

    Science.gov (United States)

    Kim, Soo-Kyoung; Park, Ha-Young; Lee, Joon-Hee

    2015-04-01

    Anthranilate and indole are alternative degradation products of tryptophan, depending on the bacterial species. While indole enhances the biofilm formation of Pseudomonas aeruginosa, we found that anthranilate, the tryptophan degradation product of P. aeruginosa, had an opposite effect on P. aeruginosa biofilm formation, in which anthranilate deteriorated the mushroom structure of biofilm. The anthranilate effect on biofilm formation was differentially exerted depending on the developmental stage and the presence of shear force. Anthranilate slightly accelerated the initial attachment of P. aeruginosa at the early stage of biofilm development and appeared to build more biofilm without shear force. But anthranilate weakened the biofilm structure in the late stage, deteriorating the mushroom structure of biofilms with shear force to make a flat biofilm. To investigate the interplay of anthranilate with indole in biofilm formation, biofilms were cotreated with anthranilate and indole, and the results showed that anthranilate antagonized the biofilm-enhancing effect of indole. Anthranilate was able to deteriorate the preformed biofilm. The effect of anthranilate and indole on biofilm formation was quorum sensing independent. AntR, a regulator of anthranilate-degrading metabolism was synergistically activated by cotreatment with anthranilate and indole, suggesting that indole might enhance biofilm formation by facilitating the degradation of anthranilate. Anthranilate slightly but significantly affected the cyclic diguaniylate (c-di-GMP) level and transcription of major extracellular polysaccharide (Psl, Pel, and alginate) operons. These results suggest that anthranilate may be a promising antibiofilm agent and antagonize the effect of indole on P. aeruginosa biofilm formation.

  5. Indole is an inter-species biofilm signal mediated by SdiA

    Directory of Open Access Journals (Sweden)

    Wood Thomas K

    2007-05-01

    Full Text Available Abstract Background As a stationary phase signal, indole is secreted in large quantities into rich medium by Escherichia coli and has been shown to control several genes (e.g., astD, tnaB, gabT, multi-drug exporters, and the pathogenicity island of E. coli; however, its impact on biofilm formation has not been well-studied. Results Through a series of global transcriptome analyses, confocal microscopy, isogenic mutants, and dual-species biofilms, we show here that indole is a non-toxic signal that controls E. coli biofilms by repressing motility, inducing the sensor of the quorum sensing signal autoinducer-1 (SdiA, and influencing acid resistance (e.g., hdeABD, gadABCEX. Isogenic mutants showed these associated proteins are directly related to biofilm formation (e.g., the sdiA mutation increased biofilm formation 50-fold, and SdiA-mediated transcription was shown to be influenced by indole. The reduction in motility due to indole addition results in the biofilm architecture changing from scattered towers to flat colonies. Additionally, there are 12-fold more E. coli cells in dual-species biofilms grown in the presence of Pseudomonas cells engineered to express toluene o-monooxygenase (TOM, which converts indole to an insoluble indigoid than in biofilms with pseudomonads that do not express TOM due to a 22-fold reduction in extracellular indole. Also, indole stimulates biofilm formation in pseudomonads. Further evidence that the indole effects are mediated by SdiA and homoserine lactone quorum sensing is that the addition of N-butyryl-, N-hexanoyl-, and N-octanoyl-L-homoserine lactones repress E. coli biofilm formation in the wild-type strain but not with the sdiA mutant. Conclusion Indole is an interspecies signal that decreases E. coli biofilms through SdiA and increases those of pseudomonads. Indole may be manipulated to control biofilm formation by oxygenases of bacteria that do not synthesize it in a dual-species biofilm. Furthermore, E

  6. pH landscapes in a novel five-species model of early dental biofilm.

    Directory of Open Access Journals (Sweden)

    Sebastian Schlafer

    Full Text Available BACKGROUND: Despite continued preventive efforts, dental caries remains the most common disease of man. Organic acids produced by microorganisms in dental plaque play a crucial role for the development of carious lesions. During early stages of the pathogenetic process, repeated pH drops induce changes in microbial composition and favour the establishment of an increasingly acidogenic and aciduric microflora. The complex structure of dental biofilms, allowing for a multitude of different ecological environments in close proximity, remains largely unexplored. In this study, we designed a laboratory biofilm model that mimics the bacterial community present during early acidogenic stages of the caries process. We then performed a time-resolved microscopic analysis of the extracellular pH landscape at the interface between bacterial biofilm and underlying substrate. METHODOLOGY/PRINCIPAL FINDINGS: Strains of Streptococcus oralis, Streptococcus sanguinis, Streptococcus mitis, Streptococcus downei and Actinomyces naeslundii were employed in the model. Biofilms were grown in flow channels that allowed for direct microscopic analysis of the biofilms in situ. The architecture and composition of the biofilms were analysed using fluorescence in situ hybridization and confocal laser scanning microscopy. Both biofilm structure and composition were highly reproducible and showed similarity to in-vivo-grown dental plaque. We employed the pH-sensitive ratiometric probe C-SNARF-4 to perform real-time microscopic analyses of the biofilm pH in response to salivary solutions containing glucose. Anaerobic glycolysis in the model biofilms created a mildly acidic environment. Decrease in pH in different areas of the biofilms varied, and distinct extracellular pH-microenvironments were conserved over several hours. CONCLUSIONS/SIGNIFICANCE: The designed biofilm model represents a promising tool to determine the effect of potential therapeutic agents on biofilm growth

  7. Biofilm-Forming Abilities of Listeria monocytogenes Serotypes Isolated from Different Sources.

    Directory of Open Access Journals (Sweden)

    Swapnil P Doijad

    Full Text Available A total of 98 previously characterized and serotyped L. monocytogenes strains, comprising 32 of 1/2a; 20 of 1/2b and 46 of 4b serotype, from clinical and food sources were studied for their capability to form a biofilm. The microtiter plate assay revealed 62 (63.26% strains as weak, 27 (27.55% strains as moderate, and 9 (9.18% strains as strong biofilm formers. Among the strong biofilm formers, 6 strains were of serotype 1/2a and 3 strains were of serotype 1/2b. None of the strain from 4b serotype exhibited strong biofilm formation. No firm correlation (p = 0.015 was noticed between any serotype and respective biofilm formation ability. Electron microscopic studies showed that strong biofilm forming isolates could synthesize a biofilm within 24 h on surfaces important in food industries such as stainless steel, ceramic tiles, high-density polyethylene plastics, polyvinyl chloride pipes, and glass. Cell enumeration of strong, moderate, and weak biofilm was performed to determine if the number of cells correlated with the biofilm-forming capabilities of the isolates. Strong, moderate, and weak biofilm showed 570±127× 103 cells/cm2, 33±26× 103 cells/cm2, 5±3× 103 cells/cm2, respectively, indicating that the number of cells was directly proportional to the strength of the biofilm. The hydrophobicity index (HI analysis revealed higher hydrophobicity with an increased biofilm formation. Fatty acid methyl esterase analysis revealed the amount of certain fatty acids such as iso-C15:0, anteiso-C15:0, and anteiso-C17:0 fatty acids correlated with the biofilm-forming capability of L. monocytogenes. This study showed that different strains of L. monocytogenes form biofilm of different intensities which did not completely correlate with their serotype; however, it correlated with the number of cells, hydrophobicity, and amount of certain fatty acids.

  8. New methods for analysis of spatial distribution and coaggregation of microbial populations in complex biofilms.

    Science.gov (United States)

    Almstrand, Robert; Daims, Holger; Persson, Frank; Sörensson, Fred; Hermansson, Malte

    2013-10-01

    In biofilms, microbial activities form gradients of substrates and electron acceptors, creating a complex landscape of microhabitats, often resulting in structured localization of the microbial populations present. To understand the dynamic interplay between and within these populations, quantitative measurements and statistical analysis of their localization patterns within the biofilms are necessary, and adequate automated tools for such analyses are needed. We have designed and applied new methods for fluorescence in situ hybridization (FISH) and digital image analysis of directionally dependent (anisotropic) multispecies biofilms. A sequential-FISH approach allowed multiple populations to be detected in a biofilm sample. This was combined with an automated tool for vertical-distribution analysis by generating in silico biofilm slices and the recently developed Inflate algorithm for coaggregation analysis of microbial populations in anisotropic biofilms. As a proof of principle, we show distinct stratification patterns of the ammonia oxidizers Nitrosomonas oligotropha subclusters I and II and the nitrite oxidizer Nitrospira sublineage I in three different types of wastewater biofilms, suggesting niche differentiation between the N. oligotropha subclusters, which could explain their coexistence in the same biofilms. Coaggregation analysis showed that N. oligotropha subcluster II aggregated closer to Nitrospira than did N. oligotropha subcluster I in a pilot plant nitrifying trickling filter (NTF) and a moving-bed biofilm reactor (MBBR), but not in a full-scale NTF, indicating important ecophysiological differences between these phylogenetically closely related subclusters. By using high-resolution quantitative methods applicable to any multispecies biofilm in general, the ecological interactions of these complex ecosystems can be understood in more detail.

  9. Failure of Standard Training Sets in the Analysis of Fast-Scan Cyclic Voltammetry Data.

    Science.gov (United States)

    Johnson, Justin A; Rodeberg, Nathan T; Wightman, R Mark

    2016-03-16

    The use of principal component regression, a multivariate calibration method, in the analysis of in vivo fast-scan cyclic voltammetry data allows for separation of overlapping signal contributions, permitting evaluation of the temporal dynamics of multiple neurotransmitters simultaneously. To accomplish this, the technique relies on information about current-concentration relationships across the scan-potential window gained from analysis of training sets. The ability of the constructed models to resolve analytes depends critically on the quality of these data. Recently, the use of standard training sets obtained under conditions other than those of the experimental data collection (e.g., with different electrodes, animals, or equipment) has been reported. This study evaluates the analyte resolution capabilities of models constructed using this approach from both a theoretical and experimental viewpoint. A detailed discussion of the theory of principal component regression is provided to inform this discussion. The findings demonstrate that the use of standard training sets leads to misassignment of the current-concentration relationships across the scan-potential window. This directly results in poor analyte resolution and, consequently, inaccurate quantitation, which may lead to erroneous conclusions being drawn from experimental data. Thus, it is strongly advocated that training sets be obtained under the experimental conditions to allow for accurate data analysis.

  10. Fast-scan Cyclic Voltammetry for the Characterization of Rapid Adenosine Release.

    Science.gov (United States)

    Nguyen, Michael D; Venton, B Jill

    2015-01-01

    Adenosine is a signaling molecule and downstream product of ATP that acts as a neuromodulator. Adenosine regulates physiological processes, such as neurotransmission and blood flow, on a time scale of minutes to hours. Recent developments in electrochemical techniques, including fast-scan cyclic voltammetry (FSCV), have allowed direct detection of adenosine with sub-second temporal resolution. FSCV studies have revealed a novel mode of rapid signaling that lasts only a few seconds. This rapid release of adenosine can be evoked by electrical or mechanical stimulations or it can be observed spontaneously without stimulation. Adenosine signaling on this time scale is activity dependent; however, the mode of release is not fully understood. Rapid adenosine release modulates oxygen levels and evoked dopamine release, indicating that adenosine may have a rapid modulatory role. In this review, we outline how FSCV can be used to detect adenosine release, compare FSCV with other techniques used to measure adenosine, and present an overview of adenosine signaling that has been characterized using FSCV. These studies point to a rapid mode of adenosine modulation, whose mechanism and function will continue to be characterized in the future.

  11. Strategies for combating bacterial biofilm infections

    DEFF Research Database (Denmark)

    Wu, Hong; Moser, Claus Ernst; Wang, Heng-Zhuang

    2015-01-01

    Formation of biofilm is a survival strategy for bacteria and fungi to adapt to their living environment, especially in the hostile environment. Under the protection of biofilm, microbial cells in biofilm become tolerant and resistant to antibiotics and the immune responses, which increases...... the difficulties for the clinical treatment of biofilm infections. Clinical and laboratory investigations demonstrated a perspicuous correlation between biofilm infection and medical foreign bodies or indwelling devices. Clinical observations and experimental studies indicated clearly that antibiotic treatment...... alone is in most cases insufficient to eradicate biofilm infections. Therefore, to effectively treat biofilm infections with currently available antibiotics and evaluate the outcomes become important and urgent for clinicians. The review summarizes the latest progress in treatment of clinical biofilm...

  12. Enzymatic removal and disinfection of bacterial biofilms

    DEFF Research Database (Denmark)

    Johansen, Charlotte; Falholt, Per; Gram, Lone

    1997-01-01

    Model biofilms of Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas fluorescens, and Pseudomonas aeruginosa were made on steel and polypropylene substrata. Plaque-resembling biofilms of Streptococcus mutans, Actinomyces, viscosus, and Fusobacterium nucleatum were made on saliva...

  13. Ciliates as engineers of phototrophic biofilms

    NARCIS (Netherlands)

    Weerman, Ellen J.; van der Geest, Harm G.; van der Meulen, Myra D.; Manders, Erik M. M.; van de Koppel, Johan; Herman, Peter M. J.; Admiraal, Wim

    2011-01-01

    1. Phototrophic biofilms consist of a matrix of phototrophs, non-photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the 'engineering' effect

  14. Candida Biofilms: Development, Architecture, and Resistance.

    Science.gov (United States)

    Chandra, Jyotsna; Mukherjee, Pranab K

    2015-08-01

    Intravascular device-related infections are often associated with biofilms (microbial communities encased within a polysaccharide-rich extracellular matrix) formed by pathogens on the surfaces of these devices. Candida species are the most common fungi isolated from catheter-, denture-, and voice prosthesis-associated infections and also are commonly isolated from contact lens-related infections (e.g., fungal keratitis). These biofilms exhibit decreased susceptibility to most antimicrobial agents, which contributes to the persistence of infection. Recent technological advances have facilitated the development of novel approaches to investigate the formation of biofilms and identify specific markers for biofilms. These studies have provided extensive knowledge of the effect of different variables, including growth time, nutrients, and physiological conditions, on biofilm formation, morphology, and architecture. In this article, we will focus on fungal biofilms (mainly Candida biofilms) and provide an update on the development, architecture, and resistance mechanisms of biofilms.

  15. Molecular Analysis of Shower Curtain Biofilm Microbes

    OpenAIRE

    Kelley, Scott T.; Theisen, Ulrike; Angenent, Largus T.; Amand, Allison St.; Pace, Norman R.

    2004-01-01

    Households provide environments that encourage the formation of microbial communities, often as biofilms. Such biofilms constitute potential reservoirs for pathogens, particularly for immune-compromised individuals. One household environment that potentially accumulates microbial biofilms is that provided by vinyl shower curtains. Over time, vinyl shower curtains accumulate films, commonly referred to as “soap scum,” which microscopy reveals are constituted of lush microbial biofilms. To dete...

  16. New Dimensions of Moving Bed Biofilm Carriers

    OpenAIRE

    Piculell, Maria

    2016-01-01

    The moving bed biofilm reactor (MBBR) is a biological wastewater treatment process in which microorganisms grow as biofilms on suspended carriers. Conventionally, MBBRs are mainly designed and optimized based on the carrier surface area, neglecting the dynamic relationship between carrier design, reactor operation and biofilm characteristics, such as biofilm thickness and the composition of the microbial community. The purpose of this research project was to learn more about the roles of the ...

  17. Comparison of the antibiotic activities of Daptomycin, Vancomycin, and the investigational Fluoroquinolone Delafloxacin against biofilms from Staphylococcus aureus clinical isolates.

    Science.gov (United States)

    Siala, Wafi; Mingeot-Leclercq, Marie-Paule; Tulkens, Paul M; Hallin, Marie; Denis, Olivier; Van Bambeke, Françoise

    2014-11-01

    Biofilm-related infections remain a scourge. In an in vitro model of biofilms using Staphylococcus aureus reference strains, delafloxacin and daptomycin were found to be the most active among the antibiotics from 8 different pharmacological classes (J. Bauer, W. Siala, P. M. Tulkens, and F. Van Bambeke, Antimicrob. Agents Chemother. 57:2726-2737, 2013, doi:10.1128/AAC.00181-13). In this study, we compared delafloxacin to daptomycin and vancomycin using biofilms produced by 7 clinical strains (S. aureus epidemic clones CC5 and CC8) in order to rationalize the differences observed between the antibiotics and strains. The effects of the antibiotics on bacterial viability (resazurin reduction assay) and biomass (crystal violet staining) were measured and correlated with the proportion of polysaccharides in the matrix, the local microenvironmental pH (micro-pH), and the antibiotic penetration in the biofilm. At clinically meaningful concentrations, delafloxacin, daptomycin, and vancomycin caused a ≥25% reduction in viability against the biofilms formed by 5, 4, and 3 strains, respectively. The antibiotic penetration within the biofilms ranged from 0.6 to 52% for delafloxacin, 0.2 to 10% for daptomycin, and 0.2 to 1% for vancomycin; for delafloxacin, this was inversely related to the polysaccharide proportion in the matrix. Six biofilms were acidic, explaining the high potency of delafloxacin (lower MICs at acidic pH). Norspermidine and norspermine (disassembling the biofilm matrix) drastically increased delafloxacin potency and efficacy (50% reduction in viability for 6 biofilms at clinically meaningful concentrations) in direct correlation with its increased penetration within the biofilm, while they only modestly improved daptomycin efficacy (50% reduction in viability for 2 biofilms) and penetration, and they showed marginal effects with vancomycin. Delafloxacin potency and efficacy against biofilms are benefited by its penetration into the matrix and the local

  18. Extracellular DNA in oral microbial biofilms.

    Science.gov (United States)

    Jakubovics, Nicholas S; Burgess, J Grant

    2015-07-01

    The extracellular matrix of microbial biofilms is critical for surface adhesion and nutrient homeostasis. Evidence is accumulating that extracellular DNA plays a number of important roles in biofilm integrity and formation on hard and soft tissues in the oral cavity. Here, we summarise recent developments in the field and consider the potential of targeting DNA for oral biofilm control.

  19. Targeting quorum sensing in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Jakobsen, Tim Holm; Bjarnsholt, Thomas; Jensen, Peter Østrup;

    2013-01-01

    Bacterial resistance to conventional antibiotics combined with an increasing acknowledgement of the role of biofilms in chronic infections has led to a growing interest in new antimicrobial strategies that target the biofilm mode of growth. In the aggregated biofilm mode, cell-to-cell communication...

  20. Biofilm Induced Tolerance Towards Antimicrobial Peptides

    DEFF Research Database (Denmark)

    Folkesson, Anders; Haagensen, Janus Anders Juul; Zampaloni, Claudia

    2008-01-01

    of microbial killing were monitored by viable count determination, and confocal laser microscopy. Strains forming structurally organized biofilms show an increased bacterial survival when challenged with colistin, compared to strains forming unstructured biofilms. The increased survival is due to genetically......, but the protection is conditional being dependent on the structural organization of the biofilm, and the induction of specific tolerance mechanisms....

  1. Confocal Microscopy Imaging of the Biofilm Matrix

    DEFF Research Database (Denmark)

    Schlafer, Sebastian; Meyer, Rikke Louise

    2016-01-01

    The extracellular matrix is an integral part of microbial biofilms and an important field of research. Confocal laser scanning microscopy is a valuable tool for the study of biofilms, and in particular of the biofilm matrix, as it allows real-time visualization of fully hydrated, living specimens...

  2. Microbiële biofilms in tandheelkunde

    NARCIS (Netherlands)

    Krom, B.P.

    2015-01-01

    Aangehechte gemeenschappen van micro-organismen, ook wel biofilms genoemd, zijn altijd en overal aanwezig. Hoewel biofilms een slechte naam hebben, zijn ze meestal natuurlijk, gezond en zelfs gewenst. In de mondzorgpraktijk komen zowel gezonde (orale biofilms) als ongezonde (bijv. in de waterleiding

  3. Microbiële biofilms in tandheelkunde

    NARCIS (Netherlands)

    Krom, B.P.

    2015-01-01

    Aangehechte gemeenschappen van micro-organismen, ook wel biofilms genoemd, zijn altijd en overal aanwezig. Hoewel biofilms een slechte naam hebben, zijn ze meestal natuurlijk, gezond en zelfs gewenst. In de tandartspraktijk komen zowel gezonde (orale biofilms) als ongezonde (bijv. in de waterleiding

  4. Magnesium limitation is an environmental trigger of the Pseudomonas aeruginosa biofilm lifestyle.

    Directory of Open Access Journals (Sweden)

    Heidi Mulcahy

    Full Text Available Biofilm formation is a conserved strategy for long-term bacterial survival in nature and during infections. Biofilms are multicellular aggregates of cells enmeshed in an extracellular matrix. The RetS, GacS and LadS sensors control the switch from a planktonic to a biofilm mode of growth in Pseudomonas aeruginosa. Here we detail our approach to identify environmental triggers of biofilm formation by investigating environmental conditions that repress expression of the biofilm repressor RetS. Mg(2+ limitation repressed the expression of retS leading to increased aggregation, exopolysaccharide (EPS production and biofilm formation. Repression of retS expression under Mg(2+ limitation corresponded with induced expression of the GacA-controlled small regulatory RNAs rsmZ and rsmY and the EPS biosynthesis operons pel and psl. We recently demonstrated that extracellular DNA sequesters Mg(2+ cations and activates the cation-sensing PhoPQ two-component system, which leads to increased antimicrobial peptide resistance in biofilms. Here we show that exogenous DNA and EDTA, through their ability to chelate Mg(2+, promoted biofilm formation. The repression of retS in low Mg(2+ was directly controlled by PhoPQ. PhoP also directly controlled expression of rsmZ but not rsmY suggesting that PhoPQ controls the equilibrium of the small regulatory RNAs and thus fine-tunes the expression of genes in the RetS pathway. In summary, Mg(2+ limitation is a biologically relevant environmental condition and the first bonafide environmental signal identified that results in transcriptional repression of retS and promotes P. aeruginosa biofilm formation.

  5. Presence of extracellular DNA in the Candida albicans biofilm matrix and its contribution to biofilms.

    Science.gov (United States)

    Martins, Margarida; Uppuluri, Priya; Thomas, Derek P; Cleary, Ian A; Henriques, Mariana; Lopez-Ribot, José L; Oliveira, Rosário

    2010-05-01

    DNA has been described as a structural component of the extracellular matrix (ECM) in bacterial biofilms. In Candida albicans, there is a scarce knowledge concerning the contribution of extracellular DNA (eDNA) to biofilm matrix and overall structure. This work examined the presence and quantified the amount of eDNA in C. albicans biofilm ECM and the effect of DNase treatment and the addition of exogenous DNA on C. albicans biofilm development as indicators of a role for eDNA in biofilm development. We were able to detect the accumulation of eDNA in biofilm ECM extracted from C. albicans biofilms formed under conditions of flow, although the quantity of eDNA detected differed according to growth conditions, in particular with regards to the medium used to grow the biofilms. Experiments with C. albicans biofilms formed statically using a microtiter plate model indicated that the addition of exogenous DNA (>160 ng/ml) increases biofilm biomass and, conversely, DNase treatment (>0.03 mg/ml) decreases biofilm biomass at later time points of biofilm development. We present evidence for the role of eDNA in C. albicans biofilm structure and formation, consistent with eDNA being a key element of the ECM in mature C. albicans biofilms and playing a predominant role in biofilm structural integrity and maintenance.

  6. Pyrosequencing reveals a core community of anodic bacterial biofilms in bioelectrochemical systems from China

    Directory of Open Access Journals (Sweden)

    Yong eXiao

    2015-12-01

    Full Text Available Bioelectrochemical systems (BESs are promising technologies for energy and product recovery coupled with wastewater treatment, and the core microbial community in electrochemically active biofilm in BESs remains controversy. In the present study, 7 anodic communities from 6 bioelectrochemical systems in 4 labs in southeast, north and south-central of China are explored by 454 pyrosequencing. A total of 251,225 effective sequences are obtained for 7 electrochemically active biofilm samples at 3% cutoff level. While Alpha-, Beta- and Gamma-proteobacteria are the most abundant classes (averaging 16.0-17.7%, Bacteroidia and Clostridia are the two sub-dominant and commonly shared classes. Six commonly shared genera i.e. Azospira, Azospirillum, Acinetobacter, Bacteroides, Geobacter, Pseudomonas and Rhodopseudomonas dominate the electrochemically active communities and are defined as core genera. A total of 25 OTUs with average relative abundance >0.5% were selected and designated as core OTUs, and some species relating to these OTUs have been reported electrochemically active. Furthermore, cyclic voltammetry and chronoamperometry tests show that two strains from Acinetobacter guillouiae and Stappia indica, bacteria relate to two core OTUs, are electrochemically active. Using randomly selected bioelectrochemical systems, the study presented extremely diverse bacterial communities in anodic biofilms, though, we still can suggest some potential microbes for investigating the electrochemical mechanisms in bioelectrochemical systems.

  7. Set potential regulation reveals additional oxidation peaks of Geobacter sulfurreducens anodic biofilms

    KAUST Repository

    Zhu, Xiuping

    2012-08-01

    Higher current densities produced in microbial fuel cells and other bioelectrochemical systems are associated with the presence of various Geobacter species. A number of electron transfer components are involved in extracellular electron transfer by the model exoelectrogen, Geobacter sulfurreducens. It has previously been shown that 5 main oxidation peaks can be identified in cyclic voltammetry scans. It is shown here that 7 separate oxidation peaks emerged over relatively long periods of time when a larger range of set potentials was used to acclimate electroactive biofilms. The potentials of oxidation peaks obtained with G. sulfurreducens biofilms acclimated at 0.60 V (vs. Ag/AgCl) were different from those that developed at - 0.46 V, and both of their peaks were different from those obtained for biofilms incubated at - 0.30 V, 0 V, and 0.30 V. These results expand the known range of potentials for which G. sulfurreducens produces identifiable oxidation peaks that could be important for extracellular electron transfer. © 2012 Elsevier B.V.

  8. From Nanowires to Biofilms: An Exploration of Novel Mechanisms of Uranium Transformation Mediated by Geobacter Bacteria

    Energy Technology Data Exchange (ETDEWEB)

    REGUERA, GEMMA [Michigan State University

    2014-01-16

    One promising strategy for the in situ bioremediation of radioactive groundwater contaminants that has been identified by the SBR Program is to stimulate the activity of dissimilatory metal-reducing microorganisms to reductively precipitate uranium and other soluble toxic metals. The reduction of U(VI) and other soluble contaminants by Geobacteraceae is directly dependent on the reduction of Fe(III) oxides, their natural electron acceptor, a process that requires the expression of Geobacter’s conductive pili (pilus nanowires). Expression of conductive pili by Geobacter cells leads to biofilm development on surfaces and to the formation of suspended biogranules, which may be physiological closer to biofilms than to planktonic cells. Biofilm development is often assumed in the subsurface, particularly at the matrix-well screen interface, but evidence of biofilms in the bulk aquifer matrix is scarce. Our preliminary results suggest, however, that biofilms develop in the subsurface and contribute to uranium transformations via sorption and reductive mechanisms. In this project we elucidated the mechanism(s) for uranium immobilization mediated by Geobacter biofilms and identified molecular markers to investigate if biofilm development is happening in the contaminated subsurface. The results provided novel insights needed in order to understand the metabolic potential and physiology of microorganisms with a known role in contaminant transformation in situ, thus having a significant positive impact in the SBR Program and providing novel concept to monitor, model, and predict biological behavior during in situ treatments.

  9. Bioreactor plant for the reproducible building of biofilms; Bioreaktoranlage zur reproduzierbaren Bildung von Biofilmen

    Energy Technology Data Exchange (ETDEWEB)

    Scheen, J. [Dortmund Univ. (Germany). Inst. fuer Umweltforschung

    1998-10-01

    Biofilms frequently appear, they are sometimes useful and sometimes disturbing. For the investigation of biofilms, it was necessary to reproduce them in specific bioreactors. Furthermore, useful media and analytical methods have to define the influence of biocides and other substances. The gravimetric analysis of microbe-formed polysaccharides in water samples and biofilm makes it possible to recognise successful treatments. These conditions are described here. Using these instruments it was possible to build up reproducible biofilms, to test and optimise them in lab examinations. In view of the expected EU Biocide Directive, this process offers possibilities for searching and verifying the suitability of biocides and other substances. (orig.) [Deutsch] Biofilme treten vielfach, sowohl nuetzlich - als auch stoerend, auf. Fuer die Untersuchungen an Biofilmen wurde eine Bioreaktoranlage, ein synthetisches Medium und eine aussagekraeftige Analytik entwickelt. Mit der gravimetrischen Messung von mikrobiell gebildeten Exopolysacchariden, aus der waessrigen Phase und direkt aus dem Biofilm heraus, konnte die Bildung von Biofilmen verfolgt werden. Mit diesem Instrumentarium war es moeglich, Biofilme reproduzierbar zu bilden und die nuetzlichen als auch schaedlichen Eigenschaften von Biofilmen im Labor zu testen und zu optimieren. Dadurch koennen Foerder- als auch Vermeidungsmethoden durch z.B. Biozide oder andere Wirkstoffe besser beurteilt werden. Im Hinblick auf die zu erwartende EU-Biozid-Richtlinie bieten sich guenstige Untersuchungsmoeglichkeiten von Bioziden auf ihre Praxistauglichkeit an. (orig.)

  10. Effects of biofilm on flow over and through a permeable bed

    Science.gov (United States)

    Kazemifar, Farzan; Blois, Gianluca; Aybar, Marcelo; Perez-Calleja, Patricia; Nerenberg, Robert; Sinha, Sumit; Hardy, Richard; Best, James; Sambrook-Smith, Gregory; Christensen, Kenneth

    2016-11-01

    Biofilms constitute an important form of bacterial life in aquatic environments and are present at the interface of fluids and solids, such as riverbeds. Biofilms are permeable, heterogeneous, and deformable structures that can influence the flow and mass/momentum transport, yet their interaction with flow is not fully understood in part due to technical obstacles impeding quantitative experimental investigations. The porosity of river beds results in the generation of a diverse mosaic of 'suction' and 'ejection' events that are far removed from typical assumptions of turbulent flow structure over an impermeable bed. In this work, the effect of biofilm on bed permeability is studied. Experiments are conducted in a closed water channel equipped with 4-cm-deep permeable bed models consisting of horizontal cylinders normal to the bulk flow direction, forming an idealized two-dimensional permeable bed. Prior to conducting flow experiments, the models are placed within an independent biofilm reactor to initiate and control the biofilm growth. Once a targeted biofilm growth stage is achieved, the models are transferred to the water channel and subjected to transitional and turbulent flows. Long-distance microscopic particle image velocimetry measurements are performed to quantify the effect of biofilm on the turbulence structure of the free flow as well as the freestream-subsurface flow interaction.

  11. Microscale geochemical gradients in Hanford 300 Area sediment biofilms and influence of uranium.

    Science.gov (United States)

    Nguyen, Hung Duc; Cao, Bin; Mishra, Bhoopesh; Boyanov, Maxim I; Kemner, Kenneth M; Fredrickson, Jim K; Beyenal, Haluk

    2012-01-01

    The presence and importance of microenvironments in the subsurface at contaminated sites were suggested by previous geochemical studies. However, no direct quantitative characterization of the geochemical microenvironments had been reported. We quantitatively characterized microscale geochemical gradients (dissolved oxygen (DO), H(2), pH, and redox potential) in Hanford 300A subsurface sediment biofilms. Our results revealed significant differences in geochemical parameters across the sediment biofilm/water interface in the presence and absence of U(VI) under oxic and anoxic conditions. While the pH was relatively constant within the sediment biofilm, the redox potential and the DO and H(2) concentrations were heterogeneous at the microscale (<500-1000 μm). We found microenvironments with high DO levels (DO hotspots) when the sediment biofilm was exposed to U(VI). On the other hand, we found hotspots (high concentrations) of H(2) under anoxic conditions both in the presence and in the absence of U(VI). The presence of anoxic microenvironments inside the sediment biofilms suggests that U(VI) reduction proceeds under bulk oxic conditions. To test this, we operated our biofilm reactor under air-saturated conditions in the presence of U(VI) and characterized U speciation in the sediment biofilm. U L(III)-edge X-ray absorption spectroscopy (XANES and EXAFS) showed that 80-85% of the U was in the U(IV) valence state.

  12. A Radio Frequency Electric Current Enhances Antibiotic Efficacy against Bacterial Biofilms

    Science.gov (United States)

    Caubet, R.; Pedarros-Caubet, F.; Chu, M.; Freye, E.; de Belém Rodrigues, M.; Moreau, J. M.; Ellison, W. J.

    2004-01-01

    Bacterial biofilms are notably resistant to antibiotic prophylaxis. The concentration of antibiotic necessary to significantly reduce the number of bacteria in the biofilm matrix can be several hundred times the MIC for the same bacteria in a planktonic phase. It has been observed that the addition of a weak continuous direct electric current to the liquid surrounding the biofilm can dramatically increase the efficacy of the antibiotic. This phenomenon, known as the bioelectric effect, has only been partially elucidated, and it is not certain that the electrical parameters are optimal. We confirm here the bioelectric effect for Escherichia coli biofilms treated with gentamicin and with oxytetracycline, and we report a new bioelectric effect with a radio frequency alternating electric current (10 MHz) instead of the usual direct current. None of the proposed explanations (transport of ions within the biofilm, production of additional biocides by electrolysis, etc.) of the direct current bioelectric effect are applicable to the radio frequency bioelectric effect. We suggest that this new phenomenon may be due to a specific action of the radio frequency electromagnetic field upon the polar parts of the molecules forming the biofilm matrix. PMID:15561841

  13. Resistance and recovery of river biofilms receiving short pulses of Triclosan and Diuron.

    Science.gov (United States)

    Proia, L; Morin, S; Peipoch, M; Romaní, A M; Sabater, S

    2011-08-01

    The effects of the herbicide Diuron (DIU) and the bactericide Triclosan (TCS) were assessed on laboratory-grown stream biofilms. Four week-old biofilms were exposed in mesocosms to 48-hours of short pulses of either DIU or TCS. The direct and indirect effects of each toxicant on the biofilms, and the subsequent recovery of the biofilms, were evaluated according to structural and functional biomarkers. These parameters were analyzed immediately before exposure, immediately after exposure, and 9 and 16days post-exposure. DIU caused an increase in diatom mortality (+79%), which persisted until the end of the experiment. TCS also affected diatom mortality (+41%), although the effect did not appear until 1week post-exposure. TCS caused an increase in bacterial mortality (+45%); however, this parameter returned to normal values 1week post-exposure. TCS compromised the cellular integrity of the green alga Spirogyra sp., whereas DIU did not. TCS also strongly inhibited phosphate uptake (-71%), which did not return to normal values until 2weeks post-exposure. DIU directly affected algae, but barely affected the heterotrophs, whereas TCS seriously impaired bacteria (direct effect) as well as autotrophs (indirect effect). However, the biofilms recovered their normal structure and function within only a few days to a few weeks. These findings demonstrate the capacity of biofilms to cope with periodic inputs of toxicants, but also the risks associated to repeated exposure or multi-contamination in aquatic ecosystems.

  14. Pattern formation in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Parsek, Matthew R.; Tolker-Nielsen, Tim

    2008-01-01

    Bacteria are capable of forming elaborate multicellular communities called biofilms. Pattern formation in biofilms depends on cell proliferation and cellular migration in response to the available nutrients and other external cues, as well as on self-generated intercellular signal molecules...... and the production of an extracellular matrix that serves as a structural 'scaffolding' for the biofilm cells. Pattern formation in biofilms allows cells to position themselves favorably within nutrient gradients and enables buildup and maintenance of physiologically distinct subpopulations, which facilitates...... survival of one or more subpopulations upon environmental insult, and therefore plays an important role in the innate tolerance displayed by biofilms toward adverse conditions....

  15. The roles of biofilm matrix polysaccharide Psl in mucoid Pseudomonas aeruginosa biofilms.

    Science.gov (United States)

    Ma, Luyan; Wang, Shiwei; Wang, Di; Parsek, Matthew R; Wozniak, Daniel J

    2012-07-01

    The opportunistic pathogen Pseudomonas aeruginosa causes life-threatening, persistent infections in patients with cystic fibrosis (CF). Persistence is attributed to the ability of these bacteria to form structured communities (biofilms). Biofilms rely on an extracellular polymeric substances matrix to maintain structure. Psl exopolysaccharide is a key matrix component of nonmucoid biofilms, yet the role of Psl in mucoid biofilms is unknown. In this report, using a variety of mutants in a mucoid P. aeruginosa background, we found that deletion of Psl-encoding genes dramatically decreased their biofilm formation ability, indicating that Psl is also a critical matrix component of mucoid biofilms. Our data also suggest that the overproduction of alginate leads to mucoid biofilms, which occupy more space, whereas Psl-dependent biofilms are densely packed. These data suggest that Psl polysaccharide may have significant contributions in biofilm persistence in patients with CF and may be helpful for designing therapies for P. aeruginosa CF infection.

  16. Combining Biofilm-Controlling Compounds and Antibiotics as a Promising New Way to Control Biofilm Infections

    Directory of Open Access Journals (Sweden)

    Andréia Bergamo Estrela

    2010-05-01

    Full Text Available Many bacteria grow on surfaces forming biofilms. In this structure, they are well protected and often high dosages of antibiotics cannot clear infectious biofilms. The formation and stabilization of biofilms are mediated by diffusible autoinducers (e.g. N-acyl homoserine lactones, small peptides, furanosyl borate diester. Metabolites interfering with this process have been identified in plants, animals and microbes, and synthetic analogues are known. Additionally, this seems to be not the only way to control biofilms. Enzymes capable of cleaving essential components of the biofilm matrix, e.g. polysaccharides or extracellular DNA, and thus weakening the biofilm architecture have been identified. Bacteria also have mechanisms to dissolve their biofilms and return to planktonic lifestyle. Only a few compounds responsible for the signalling of these processes are known, but they may open a completely novel line of biofilm control. All these approaches lead to the destruction of the biofilm but not the killing of the pathogens. Therefore, a combination of biofilm-destroying compounds and antibiotics to handle biofilm infections is proposed. In this article, different approaches to combine biofilm-controlling compounds and antibiotics to fight biofilm infections are discussed, as well as the balance between biofilm formation and virulence.

  17. Vibrio cholerae Biofilms and Cholera Pathogenesis.

    Directory of Open Access Journals (Sweden)

    Anisia J Silva

    2016-02-01

    Full Text Available Vibrio cholerae can switch between motile and biofilm lifestyles. The last decades have been marked by a remarkable increase in our knowledge of the structure, regulation, and function of biofilms formed under laboratory conditions. Evidence has grown suggesting that V. cholerae can form biofilm-like aggregates during infection that could play a critical role in pathogenesis and disease transmission. However, the structure and regulation of biofilms formed during infection, as well as their role in intestinal colonization and virulence, remains poorly understood. Here, we review (i the evidence for biofilm formation during infection, (ii the coordinate regulation of biofilm and virulence gene expression, and (iii the host signals that favor V. cholerae transitions between alternative lifestyles during intestinal colonization, and (iv we discuss a model for the role of V. cholerae biofilms in pathogenicity.

  18. Role of multicellular aggregates in biofilm formation

    DEFF Research Database (Denmark)

    Kragh, Kasper N.; Hutchison, Jaime B.; Melaugh, Gavin;

    2016-01-01

    response, may add to this ecological benefit. Our findings suggest that current models of biofilm formation should be reconsidered to incorporate the role of aggregates in biofilm initiation.IMPORTANCE During the past decades, there has been a consensus around the model of development of a biofilm......In traditional models of in vitro biofilm development, individual bacterial cells seed a surface, multiply, and mature into multicellular, three-dimensional structures. Much research has been devoted to elucidating the mechanisms governing the initial attachment of single cells to surfaces. However......, in natural environments and during infection, bacterial cells tend to clump as multicellular aggregates, and biofilms can also slough off aggregates as a part of the dispersal process. This makes it likely that biofilms are often seeded by aggregates and single cells, yet how these aggregates impact biofilm...

  19. Modelling the growth of a methanotrophic biofilm

    DEFF Research Database (Denmark)

    Arcangeli, J.-P.; Arvin, E.

    1999-01-01

    This article discusses the growth of methanotrophic biofilms. Several independent biofilm growths scenarios involving different inocula were examined. Biofilm growth, substrate removal and product formation were monitored throughout the experiments. Based on the oxygen consumption it was concluded...... that heterotrophs and nitrifiers co-existed with methanotrophs in the biofilm. Heterotrophic biomass grew on soluble polymers formed by the hydrolysis of dead biomass entrapped in the biofilm. Nitrifier populations developed because of the presence of ammonia in the mineral medium. Based on these experimental...... was performed on this model. It indicated that the most influential parameters were those related to the biofilm (i.e. density; solid-volume fraction; thickness). This suggests that in order to improve the model, further research regarding the biofilm structure and composition is needed....

  20. Molecular Basis for Saccharomyces cerevisiae Biofilm Development

    DEFF Research Database (Denmark)

    Andersen, Kaj Scherz

    of translation of FLO11. In conclusion, I have conducted the first global study of the genetic program for yeast biofilm formation on polystyrene. This work provide several target genes as good basis for further research of biofilm, that I believe can contribute to fields such as cell biology, genetics, system......In this study, I sought to identify genes regulating the global molecular program for development of sessile multicellular communities, also known as biofilm, of the eukaryotic microorganism, Saccharomyces cerevisiae (yeast). Yeast biofilm has a clinical interest, as biofilms can cause chronic......, but only a small subset is previously described as regulators of FLO11. These results reveal that the regulation of biofilm formation and FLO11 is even more complex than what has previously been described. I find that the molecular program for biofilm formation shares many essential components with two...

  1. 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....

  2. The interconnection between biofilm formation and horizontal gene transfer.

    Science.gov (United States)

    Madsen, Jonas Stenløkke; Burmølle, Mette; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

    2012-07-01

    Recent research has revealed that horizontal gene transfer and biofilm formation are connected processes. Although published research investigating this interconnectedness is still limited, we will review this subject in order to highlight the potential of these observations because of their believed importance in the understanding of the adaptation and subsequent evolution of social traits in bacteria. Here, we discuss current evidence for such interconnectedness centred on plasmids. Horizontal transfer rates are typically higher in biofilm communities compared with those in planktonic states. Biofilms, furthermore, promote plasmid stability and may enhance the host range of mobile genetic elements that are transferred horizontally. Plasmids, on the other hand, are very well suited to promote the evolution of social traits such as biofilm formation. This, essentially, transpires because plasmids are independent replicons that enhance their own success by promoting inter-bacterial interactions. They typically also carry genes that heighten their hosts' direct fitness. Furthermore, current research shows that the so-called mafia traits encoded on mobile genetic elements can enforce bacteria to maintain stable social interactions. It also indicates that horizontal gene transfer ultimately enhances the relatedness of bacteria carrying the mobile genetic elements of the same origin. The perspective of this review extends to an overall interconnectedness between horizontal gene transfer, mobile genetic elements and social evolution of bacteria.

  3. Biofilm monitoring using complex permittivity.

    Energy Technology Data Exchange (ETDEWEB)

    Altman, Susan Jeanne; McGrath, Lucas K.; Dolan, Patricia L.; Yelton, William Graham

    2008-10-01

    There is strong interest in the detection and monitoring of bio-fouling. Bio-fouling problems are common in numerous water treatments systems, medical and dental apparatus and food processing equipment. Current bio-fouling control protocols are time consuming and costly. New early detection techniques to monitor bio-forming contaminates are means to enhanced efficiency. Understanding the unique dielectric properties of biofilm development, colony forming bacteria and nutrient background will provide a basis to the effectiveness of controlling or preventing biofilm growth. Dielectric spectroscopy measurements provide values of complex permittivity, {var_epsilon}*, of biofilm formation by applying a weak alternating electric field at various frequencies. The dielectric characteristic of the biofilm, {var_epsilon}{prime}, is the real component of {var_epsilon}* and measures the biofilm's unique ability to store energy. Graphically observed dependencies of {var_epsilon}{prime} to frequency indicate dielectric relaxation or dielectric dispersion behaviors that mark the particular stage of progression during the development of biofilms. In contrast, any frequency dependency of the imaginary component, {var_epsilon}{double_prime} the loss factor, is expressed as dielectric losses from the biofilm due to dipole relaxation. The tangent angle of these two component vectors is the ratio of the imaginary component to the real component, {var_epsilon}{double_prime}/{var_epsilon}{prime} and is referred to as the loss angle tangent (tan {delta}) or dielectric loss. Changes in tan {delta} are characteristic of changes in dielectric losses during various developmental stages of the films. Permittivity scans in the above figure are of biofilm growth from P. Fluorescens (10e7 CFU's at the start). Three trends are apparent from these scans, the first being a small drop in the imaginary permittivity over a 7 hours period, best seen in the Cole-Cole plot (a). The second trend

  4. Functional Divergence of Hsp90 Genetic Interactions in Biofilm and Planktonic Cellular States.

    Directory of Open Access Journals (Sweden)

    Stephanie Diezmann

    Full Text Available Candida albicans is among the most prevalent opportunistic fungal pathogens. Its capacity to cause life-threatening bloodstream infections is associated with the ability to form biofilms, which are intrinsically drug resistant reservoirs for dispersal. A key regulator of biofilm drug resistance and dispersal is the molecular chaperone Hsp90, which stabilizes many signal transducers. We previously identified 226 C. albicans Hsp90 genetic interactors under planktonic conditions, of which 56 are involved in transcriptional regulation. Six of these transcriptional regulators have previously been implicated in biofilm formation, suggesting that Hsp90 genetic interactions identified in planktonic conditions may have functional significance in biofilms. Here, we explored the relationship between Hsp90 and five of these transcription factor genetic interactors: BCR1, MIG1, TEC1, TUP1, and UPC2. We deleted each transcription factor gene in an Hsp90 conditional expression strain, and assessed biofilm formation and morphogenesis. Strikingly, depletion of Hsp90 conferred no additional biofilm defect in the mutants. An interaction was observed in which deletion of BCR1 enhanced filamentation upon reduction of Hsp90 levels. Further, although Hsp90 modulates expression of TEC1, TUP1, and UPC2 in planktonic conditions, it has no impact in biofilms. Lastly, we probed for physical interactions between Hsp90 and Tup1, whose WD40 domain suggests that it might interact with Hsp90 directly. Hsp90 and Tup1 formed a stable complex, independent of temperature or developmental state. Our results illuminate a physical interaction between Hsp90 and a key transcriptional regulator of filamentation and biofilm formation, and suggest that Hsp90 has distinct genetic interactions in planktonic and biofilm cellular states.

  5. Degradation in the dentin-composite interface subjected to multi-species biofilm challenges.

    Science.gov (United States)

    Li, Y; Carrera, C; Chen, R; Li, J; Lenton, P; Rudney, J D; Jones, R S; Aparicio, C; Fok, A

    2014-01-01

    Oral biofilms can degrade the components in dental resin-based composite restorations, thus compromising marginal integrity and leading to secondary caries. This study investigates the mechanical integrity of the dentin-composite interface challenged with multi-species oral biofilms. While most studies used single-species biofilms, the present study used a more realistic, diverse biofilm model produced directly from plaques collected from donors with a history of early childhood caries. Dentin-composite disks were made using bovine incisor roots filled with Z100(TM) or Filtek(TM) LS (3M ESPE). The disks were incubated for 72 h in paired CDC biofilm reactors, using a previously published protocol. One reactor was pulsed with sucrose, and the other was not. A sterile saliva-only control group was run with sucrose pulsing. The disks were fractured under diametral compression to evaluate their interfacial bond strength. The surface deformation of the disks was mapped using digital image correlation to ascertain the fracture origin. Fracture surfaces were examined using scanning electron microscopy/energy-dispersive X-ray spectroscopy to assess demineralization and interfacial degradation. Dentin demineralization was greater under sucrose-pulsed biofilms, as the pH dropped Biofilm growth with sucrose pulsing also caused preferential degradation of the composite-dentin interface, depending on the composite/adhesive system used. Specifically, Z100 specimens showed greater bond strength reduction and more frequent cohesive failure in the adhesive layer. This was attributed to the inferior dentin coverage by Z100 adhesive, which possibly led to a higher level of chemical and enzymatic degradation. The results suggested that factors other than dentin demineralization were also responsible for interfacial degradation. A clinically relevant in vitro biofilm model was therefore developed, which would effectively allow assessment of the degradation of the dentin

  6. Biofilm feeding: Microbial colonization of food promotes the growth of a detritivorous arthropod

    Science.gov (United States)

    Horváthová, Terézia; Babik, Wiesław; Bauchinger, Ulf

    2016-01-01

    Abstract Feeding on plant material is common among animals, but how different animals overcome the dietary deficiencies imposed by this feeding strategy is not well understood. Microorganisms are generally considered to play a vital role in the nutritional ecology of plant feeding animals. Commonly microbes living inside animal bodies are considered more important, but recent studies suggest external microbes significantly shape plant-feeding strategies in invertebrates. Here we investigate how external microbes that typically form biofilm on primary plant material affect growth rates in a terrestrial isopod species Porcellio scaber. We experimentally manipulated the amount of biofilm on three different primary diet sources and quantified growth and survival of individuals that fed on food with either a small or large amount of biofilm. In addition, we tested how dietary manipulation shapes the composition of bacterial communities in the gut. The presence of visible biofilm significantly affected the growth of isopods: individuals that fed on the primary diet source with a large amount of biofilm gained more mass than individuals feeding on a diet with marginal biofilm. Diet also significantly affected the bacterial gut community. The primary diet source mainly determined the taxonomic composition of the bacterial community in the isopod gut, whereas the amount of biofilm affected the relative abundance of bacterial taxa. Our study suggests that terrestrial isopods may cope with low-quality plant matter by feeding on biofilm, with decomposition of plant material by organisms outside of the feeding organism (here a terrestrial isopod) probably playing a major role. Future investigations may be directed towards the primary diet source, plant matter, and the secondary diet source, biofilm, and should assess if both components are indeed uptaken in detritivorous species. PMID:27110187

  7. Study on Water—Soluble Organic Reducing Substances.I.Determination of Organic Reducing Substances by Differential Pulse Voltammetry

    Institute of Scientific and Technical Information of China (English)

    WUYOU-XIAN; DINGCHANG-PU

    1991-01-01

    A new method was proposed for study of organic reducing substances in soils.According to the theoretical relationship between the voltammetric behaviors and reduction-oxidation reaction of reducing substances,the working conditions of differential pulse voltammetry (d.p.v.)for determining the organic reducing substances produced during the processes of the anaerobic decomposition of plant materials were established with a glass carbon electrode as working electrode,1M Ag-AgCl electrode with large area as reference electrode,0.2M NH4Ac as supporting from -0.5 to +1.2 voltage(vs.M Ag-AgCl).The peak current proportional to the concentration of reducing substances,and the characteristic peak potential of each organic reducing substance were regarded as the quantitative and qualitative base,respectively.These results obtained under the conditions mentioned above directly reflect both the reducing intensity and capacity of the organic reducing system in soils.

  8. Rapid identification of bacterial biofilms and biofilm wound models using a multichannel nanosensor.

    Science.gov (United States)

    Li, Xiaoning; Kong, Hao; Mout, Rubul; Saha, Krishnendu; Moyano, Daniel F; Robinson, Sandra M; Rana, Subinoy; Zhang, Xinrong; Riley, Margaret A; Rotello, Vincent M

    2014-12-23

    Identification of infectious bacteria responsible for biofilm-associated infections is challenging due to the complex and heterogeneous biofilm matrix. To address this issue and minimize the impact of heterogeneity on biofilm identification, we developed a gold nanoparticle (AuNP)-based multichannel sensor to detect and identify biofilms based on their physicochemical properties. Our results showed that the sensor can discriminate six bacterial biofilms including two composed of uropathogenic bacteria. The capability of the sensor was further demonstrated through discrimination of biofilms in a mixed bacteria/mammalian cell in vitro wound model.

  9. Biofilms in chronic infections - a matter of opportunity - monospecies biofilms in multispecies infections

    DEFF Research Database (Denmark)

    Burmølle, Mette; Thomsen, Trine Rolighed; Fazli, Mustafa

    2010-01-01

    to permanent tissue fillers and chronic wounds) both as to distribution (such as where in the wound bed) and organization (monospecies or multispecies microcolonies). We correlate these biofilm observations to observations of commensal biofilms (dental and intestine) and biofilms in natural ecosystems (soil......). The observations of the chronic biofilm infections point toward a trend of low bacterial diversity and sovereign monospecies biofilm aggregates even though the infection in which they reside are multispecies. In contrast to this, commensal and natural biofilm aggregates contain multiple species that are believed...

  10. Important contribution of the novel locus comEB to extracellular DNA-dependent Staphylococcus lugdunensis biofilm formation.

    Science.gov (United States)

    Rajendran, Nithya Babu; Eikmeier, Julian; Becker, Karsten; Hussain, Muzaffar; Peters, Georg; Heilmann, Christine

    2015-12-01

    The coagulase-negative species Staphylococcus lugdunensis is an emerging cause of serious and potentially life-threatening infections, such as infective endocarditis. The pathogenesis of these infections is characterized by the ability of S. lugdunensis to form biofilms on either biotic or abiotic surfaces. To elucidate the genetic basis of biofilm formation in S. lugdunensis, we performed transposon (Tn917) mutagenesis. One mutant had a significantly reduced biofilm-forming capacity and carried a Tn917 insertion within the competence gene comEB. Site-directed mutagenesis and subsequent complementation with a functional copy of comEB verified the importance of comEB in biofilm formation. In several bacterial species, natural competence stimulates DNA release via lysis-dependent or -independent mechanisms. Extracellular DNA (eDNA) has been demonstrated to be an important structural component of many bacterial biofilms. Therefore, we quantified the eDNA in the biofilms and found diminished eDNA amounts in the comEB mutant biofilm. High-resolution images and three-dimensional data obtained via confocal laser scanning microscopy (CSLM) visualized the impact of the comEB mutation on biofilm integrity. The comEB mutant did not show reduced expression of autolysin genes, decreased autolytic activities, or increased cell viability, suggesting a cell lysis-independent mechanism of DNA release. Furthermore, reduced amounts of eDNA in the comEB mutant biofilms did not result from elevated levels or activity of the S. lugdunensis thermonuclease NucI. In conclusion, we defined here, for the first time, a role for the competence gene comEB in staphylococcal biofilm formation. Our findings indicate that comEB stimulates biofilm formation via a lysis-independent mechanism of DNA release.

  11. Diagnostic criteria for the characterization of quasireversible electron transfer reactions by cyclic square wave voltammetry.

    Science.gov (United States)

    Mann, Megan A; Helfrick, John C; Bottomley, Lawrence A

    2014-08-19

    Theory for cyclic square wave voltammetry of quasireversible electron transfer reactions is presented and experimentally verified. The impact of empirical parameters on the shape of the current-voltage curve is examined. From the trends, diagnostic criteria enabling the use of this waveform as a tool for mechanistic analysis of electrode reaction processes are presented. These criteria were experimentally confirmed using Eu(3+)/Eu(2+), a well-established quasireversible analyte. Using cyclic square wave voltammetry, both the electron transfer coefficient and rate were calculated for this analyte and found to be in excellent agreement with literature. When properly applied, these criteria will enable nonexperts in voltammetry to assign the electrode reaction mechanism and accurately measure electrode reaction kinetics.

  12. Fast voltammetry of metals at carbon-fiber microelectrodes: towards an online speciation sensor.

    Science.gov (United States)

    Pathirathna, Pavithra; Siriwardhane, Thushani; McElmurry, Shawn P; Morgan, Stephen L; Hashemi, Parastoo

    2016-11-14

    Speciation controls the chemical behavior of trace metals. Thus, there is great demand for rapid speciation analysis in a variety of fields. In this study, we describe the application of fast scan cyclic voltammetry (FSCV) and fast scan adsorption controlled voltammetry (FSCAV) to trace metal speciation analysis. We show that Cu(2+) can be detected using FSCAV in different matrices. We find that matrices with different Cu(2+) binding ability do not affect the equilibrium of Cu(2+) adsorption onto CFMs, and thus are an excellent predictor for free Cu(2+) ([Cu(2+)]free) in solution. We modelled a correlation between the FSCV response, [Cu(2+)]free and log Kf for 15 different Cu(2+) complexes. Using our model, we rapidly predicted, and verified [Cu(2+)]free and Kf of a real groundwater sample spiked with Cu(2+). We thus highlight the potential of fast voltammetry as a rapid trace metal speciation sensor.

  13. Linear surface roughness growth and flow smoothening in a three-dimensional biofilm model

    Science.gov (United States)

    Head, D. A.

    2013-09-01

    The sessile microbial communities known as biofilms exhibit varying architectures as environmental factors are varied, which for immersed biofilms includes the shear rate of the surrounding flow. Here we modify an established agent-based biofilm model to include affine flow and employ it to analyze the growth of surface roughness of single-species, three-dimensional biofilms. We find linear growth laws for surface geometry in both horizontal and vertical directions and measure the thickness of the active surface layer, which is shown to anticorrelate with roughness. Flow is shown to monotonically reduce surface roughness without affecting the thickness of the active layer. We argue that the rapid roughening is due to nonlocal surface interactions mediated by the nutrient field, which are curtailed when advection competes with diffusion. We further argue the need for simplified models to elucidate the underlying mechanisms coupling flow to growth.

  14. Electrochemical impedance spectroscopy of biofilms

    Science.gov (United States)

    Microbial activity that leads to the formation of biofilms on process equipment can accelerate corrosion, reduce heat transfer rates, and generally decrease process efficiencies. Additional concerns arise in the food and pharma industries where product quality and safety are a high priority. Pharmac...

  15. Exploiting social evolution in biofilms

    DEFF Research Database (Denmark)

    Boyle, Kerry E; Heilmann, Silja; van Ditmarsch, Dave

    2013-01-01

    and thus, regrettably, select for resistance against their own action. A possible solution lies in targeting the mechanisms by which bacteria interact with each other within biofilms. The emerging field of microbial social evolution combines molecular microbiology with evolutionary theory to dissect...

  16. Biogenesis of Enterococcis faecium biofilms

    NARCIS (Netherlands)

    Paganelli, F.L.

    2015-01-01

    Nosocomial infections caused by Enterococcus faecium have rapidly increased worldwide and treatment options become more limited. The presence of antibiotic resistance genes and virulence factors in pathogenic E. faecium contribute to difficult-to-treat infections, frequently biofilm mediated, such a

  17. Biofilm Formation and Morphotypes of Salmonella enterica subsp.arizonae Differs from Those of Other Salmonella enterica Subspecies in Isolates from Poultry Houses.

    Science.gov (United States)

    Lamas, A; Fernandez-No, I C; Miranda, J M; Vázquez, B; Cepeda, A; Franco, C M

    2016-07-01

    Salmonella serovars are responsible for foodborne diseases around the world. The ability to form biofilms allows microorganisms to survive in the environment. In this study, 73 Salmonella strains, belonging to four different subspecies, were isolated from poultry houses and foodstuffs and tested. Biofilm formation was measured at four different temperatures and two nutrient concentrations. Morphotypes and cellulose production were evaluated at three different temperatures. The presence of several genes related to biofilm production was also examined. All strains and subspecies of Salmonella had the ability to form biofilms, and 46.57% of strains produced biofilms under all conditions tested. Biofilm formation was strain dependent and varied according to the conditions. This is the first study to analyze biofilm formation in a wide number of Salmonella enterica subsp. arizonae strains, and no direct relationship between the high prevalence of Salmonella enterica subsp. arizonae strains and their ability to form biofilm was established. Morphotypes and cellulose production varied as the temperature changed, with 20°C being the optimum temperature for expression of the red, dry, and rough morphotype and cellulose. Salmonella enterica subsp. arizonae, whose morphotype is poorly studied, only showed a smooth and white morphotype and lacked the csgD and gcpA genes that are implicated in biofilm production. Thus, Salmonella biofilm formation under different environmental conditions is a public health problem because it can survive and advance through the food chain to reach the consumer.

  18. Anodic passivation of tin by alkanethiol self-assembled monolayers examined by cyclic voltammetry and coulometry.

    Science.gov (United States)

    Worley, Barrett C; Ricks, William A; Prendergast, Michael P; Gregory, Brian W; Collins, Ross; Cassimus, John J; Thompson, Raymond G

    2013-10-22

    The self-assembly of medium chain length alkanethiol monolayers on polycrystalline Sn electrodes has been investigated by cyclic voltammetry and coulometry. These studies have been performed in order to ascertain the conditions under which their oxidative deposition can be achieved directly on the oxide-free Sn surface, and the extent to which these electrochemically prepared self-assembled monolayers (SAMs) act as barriers to surface oxide growth. This work has shown that the potentials for their oxidative deposition are more cathodic (by 100-200 mV) than those for Sn surface oxidation and that the passivating abilities of these SAMs improve with increasing film thickness (or chain length). Oxidative desorption potentials for these films have been observed to shift more positively, and in a highly linear fashion, with increasing film thickness (~75 mV/CH2). Although reductive desorption potentials for the SAMs are in close proximity to those for reduction of the surface oxide (SnOx), little or no SnOx formation occurs unless the potential is made sufficiently anodic that the monolayers start to be removed oxidatively. Our coulometric data indicate that the charge involved with alkanethiol reductive desorption or oxidative deposition is consistent with the formation of a close-packed monolayer, given uncertainties attributable to surface roughness and heterogeneity phenomena. These experiments also reveal that the quantity of charge passed during oxidative desorption is significantly larger than what would be predicted for simple alkylsulfinate or alkylsulfonate formation, suggesting that oxidative removal involves a more complex oxidation mechanism. Analogous chronocoulometric experiments for short-chain alkanethiols on polycrystalline Au electrodes have evidenced similar oxidative charge densities. This implies that the mechanism for oxidative desorption on both surfaces may be very similar, despite the significant differences in the inherent dissolution

  19. Hydrodynamic dispersion within porous biofilms

    KAUST Repository

    Davit, Y.

    2013-01-23

    Many microorganisms live within surface-associated consortia, termed biofilms, that can form intricate porous structures interspersed with a network of fluid channels. In such systems, transport phenomena, including flow and advection, regulate various aspects of cell behavior by controlling nutrient supply, evacuation of waste products, and permeation of antimicrobial agents. This study presents multiscale analysis of solute transport in these porous biofilms. We start our analysis with a channel-scale description of mass transport and use the method of volume averaging to derive a set of homogenized equations at the biofilm-scale in the case where the width of the channels is significantly smaller than the thickness of the biofilm. We show that solute transport may be described via two coupled partial differential equations or telegrapher\\'s equations for the averaged concentrations. These models are particularly relevant for chemicals, such as some antimicrobial agents, that penetrate cell clusters very slowly. In most cases, especially for nutrients, solute penetration is faster, and transport can be described via an advection-dispersion equation. In this simpler case, the effective diffusion is characterized by a second-order tensor whose components depend on (1) the topology of the channels\\' network; (2) the solute\\'s diffusion coefficients in the fluid and the cell clusters; (3) hydrodynamic dispersion effects; and (4) an additional dispersion term intrinsic to the two-phase configuration. Although solute transport in biofilms is commonly thought to be diffusion dominated, this analysis shows that hydrodynamic dispersion effects may significantly contribute to transport. © 2013 American Physical Society.

  20. Reinterpretation of reduction potential measurements done by linear sweep voltammetry in silicate melts

    Science.gov (United States)

    Colson, R. O.; Haskin, L. A.; Keedy, C. R.

    1991-01-01

    The equilibrium concentrations of Ni between silicate melt and Pt were determined experimentally as a function of oxygen fugacity. The results demonstrate that metallic species derived in linear sweep voltammetry experiments in silicate melts are diffusing into Pt electrodes and not into the melt, as was concluded by previoius studies. This requires reinterpretation of previous linear sweep voltammetry results and recalculation and correction of reported reduction potentials. This paper reports these corrected reduction potentials. Also reported are the activity coefficients for Ni in synthetic basalt and diopsidic melts and for Co in diopsidic melt.

  1. Detection of gamma-irradiation effect on DNA and protein using magnetic sensor and cyclic voltammetry.

    Science.gov (United States)

    Park, Duck-Gun; Song, Hoon; Kishore, M B; Vértesy, G; Lee, Duk-Hyun

    2013-11-01

    In this study, a magnetic sensor utilizing Planar Hall Resistance (PHR) and cyclic Voltammetry (CV) for detecting the radiation effect was fabricated. Specifically, we applied in parallel a PHR sensor and CV device to monitor the irradiation effect on DNA and protein respectively. Through parallel measurements, we demonstrated that the PHR sensor and CV are sensitive enough to measure irradiation effect. The PHR voltage decreased by magnetic nanobead labeled DNA was slightly recovered after gamma ray irradiation. The behavior of cdk inhibitor protein p21 having a sandwich structure of Au/protein G/Ab/Ag/Ab was checked by monitoring the cyclic Voltammetry signal in analyzing the gamma ray irradiation effect.

  2. Chemical Biology Strategies for Biofilm Control.

    Science.gov (United States)

    Yang, Liang; Givskov, Michael

    2015-08-01

    Microbes live as densely populated multicellular surface-attached biofilm communities embedded in self-generated, extracellular polymeric substances (EPSs). EPSs serve as a scaffold for cross-linking biofilm cells and support development of biofilm architecture and functions. Biofilms can have a clear negative impact on humans, where biofilms are a common denominator in many chronic diseases in which they prime development of destructive inflammatory conditions and the failure of our immune system to efficiently cope with them. Our current assortment of antimicrobial agents cannot efficiently eradicate biofilms. For industrial applications, the removal of biofilms within production machinery in the paper and hygienic food packaging industry, cooling water circuits, and drinking water manufacturing systems can be critical for the safety and efficacy of those processes. Biofilm formation is a dynamic process that involves microbial cell migration, cell-to-cell signaling and interactions, EPS synthesis, and cell-EPS interactions. Recent progress of fundamental biofilm research has shed light on novel chemical biology strategies for biofilm control. In this article, chemical biology strategies targeting the bacterial intercellular and intracellular signaling pathways will be discussed.

  3. From Mouth to Model: Combining in vivo and in vitro Oral Biofilm Growth

    Science.gov (United States)

    Klug, Barbara; Santigli, Elisabeth; Westendorf, Christian; Tangl, Stefan; Wimmer, Gernot; Grube, Martin

    2016-01-01

    Background: Oral biofilm studies based on simplified experimental setups are difficult to interpret. Models are limited mostly by the number of bacterial species observed and the insufficiency of artificial media. Few studies have attempted to overcome these limitations and to cultivate native oral biofilm. Aims: This study aimed to grow oral biofilm in vivo before transfer to a biofilm reactor for ex situ incubation. The in vitro survival of this oral biofilm and the changes in bacterial composition over time were observed. Methods: Six human enamel-dentin slabs embedded buccally in dental splints were used as biofilm carriers. Fitted individually to the upper jaw of 25 non-smoking male volunteers, the splints were worn continuously for 48 h. During this time, tooth-brushing and alcohol-consumption were not permitted. The biofilm was then transferred on slabs into a biofilm reactor and incubated there for 48 h while being nourished in BHI medium. Live/dead staining and confocal laser scanning microscopy were used to observe bacterial survival over four points in time: directly after removal (T0) and after 1 (T1), 24 (T2), and 48 h (T3) of incubation. Bacterial diversity at T0 and T3 was compared with 454-pyrosequencing. Fluorescence in situ hybridization (FISH) was performed to show specific taxa. Survival curves were calculated with a specially designed MATLAB script. Acacia and QIIME 1.9.1 were used to process pyrosequencing data. SPSS 21.0 and R 3.3.1 were used for statistical analysis. Results: After initial fluctuations at T1, survival curves mostly showed approximation of the bacterial numbers to the initial level at T3. Pyrosequencing analysis resulted in 117 OTUs common to all samples. The genera Streptococcus and Veillonella (both Firmicutes) dominated at T0 and T3. They make up two thirds of the biofilm. Genera with lower relative abundance had grown significantly at T3. FISH analysis confirmed the pyrosequencing results, i.e., the predominant staining

  4. From Mouth to Model: Combining in vivo and in vitro Oral Biofilm Growth.

    Science.gov (United States)

    Klug, Barbara; Santigli, Elisabeth; Westendorf, Christian; Tangl, Stefan; Wimmer, Gernot; Grube, Martin

    2016-01-01

    Background: Oral biofilm studies based on simplified experimental setups are difficult to interpret. Models are limited mostly by the number of bacterial species observed and the insufficiency of artificial media. Few studies have attempted to overcome these limitations and to cultivate native oral biofilm. Aims: This study aimed to grow oral biofilm in vivo before transfer to a biofilm reactor for ex situ incubation. The in vitro survival of this oral biofilm and the changes in bacterial composition over time were observed. Methods: Six human enamel-dentin slabs embedded buccally in dental splints were used as biofilm carriers. Fitted individually to the upper jaw of 25 non-smoking male volunteers, the splints were worn continuously for 48 h. During this time, tooth-brushing and alcohol-consumption were not permitted. The biofilm was then transferred on slabs into a biofilm reactor and incubated there for 48 h while being nourished in BHI medium. Live/dead staining and confocal laser scanning microscopy were used to observe bacterial survival over four points in time: directly after removal (T0) and after 1 (T1), 24 (T2), and 48 h (T3) of incubation. Bacterial diversity at T0 and T3 was compared with 454-pyrosequencing. Fluorescence in situ hybridization (FISH) was performed to show specific taxa. Survival curves were calculated with a specially designed MATLAB script. Acacia and QIIME 1.9.1 were used to process pyrosequencing data. SPSS 21.0 and R 3.3.1 were used for statistical analysis. Results: After initial fluctuations at T1, survival curves mostly showed approximation of the bacterial numbers to the initial level at T3. Pyrosequencing analysis resulted in 117 OTUs common to all samples. The genera Streptococcus and Veillonella (both Firmicutes) dominated at T0 and T3. They make up two thirds of the biofilm. Genera with lower relative abundance had grown significantly at T3. FISH analysis confirmed the pyrosequencing results, i.e., the predominant staining

  5. Broad-spectrum anti-biofilm peptide that targets a cellular stress response.

    Directory of Open Access Journals (Sweden)

    César de la Fuente-Núñez

    2014-05-01

    Full Text Available Bacteria form multicellular communities known as biofilms that cause two thirds of all infections and demonstrate a 10 to 1000 fold increase in adaptive resistance to conventional antibiotics. Currently, there are no approved drugs that specifically target bacterial biofilms. Here we identified a potent anti-biofilm peptide 1018 that worked by blocking (pppGpp, an important signal in biofilm development. At concentrations that did not affect planktonic growth, peptide treatment completely prevented biofilm formation and led to the eradication of mature biofilms in representative strains of both Gram-negative and Gram-positive bacterial pathogens including Pseudomonas aeruginosa, Escherichia coli, Acinetobacter baumannii, Klebsiella pneumoniae, methicillin resistant Staphylococcus aureus, Salmonella Typhimurium and Burkholderia cenocepacia. Low levels of the peptide led to biofilm dispersal, while higher doses triggered biofilm cell death. We hypothesized that the peptide acted to inhibit a common stress response in target species, and that the stringent response, mediating (pppGpp synthesis through the enzymes RelA and SpoT, was targeted. Consistent with this, increasing (pppGpp synthesis by addition of serine hydroxamate or over-expression of relA led to reduced susceptibility to the peptide. Furthermore, relA and spoT mutations blocking production of (pppGpp replicated the effects of the peptide, leading to a reduction of biofilm formation in the four tested target species. Also, eliminating (pppGpp expression after two days of biofilm growth by removal of arabinose from a strain expressing relA behind an arabinose-inducible promoter, reciprocated the effect of peptide added at the same time, leading to loss of biofilm. NMR and chromatography studies showed that the peptide acted on cells to cause degradation of (pppGpp within 30 minutes, and in vitro directly interacted with ppGpp. We thus propose that 1018 targets (pppGpp and marks it for

  6. Dynamic interactions of neutrophils and biofilms

    Directory of Open Access Journals (Sweden)

    Josefine Hirschfeld

    2014-12-01

    Full Text Available Background: The majority of microbial infections in humans are biofilm-associated and difficult to treat, as biofilms are highly resistant to antimicrobial agents and protect themselves from external threats in various ways. Biofilms are tenaciously attached to surfaces and impede the ability of host defense molecules and cells to penetrate them. On the other hand, some biofilms are beneficial for the host and contain protective microorganisms. Microbes in biofilms express pathogen-associated molecular patterns and epitopes that can be recognized by innate immune cells and opsonins, leading to activation of neutrophils and other leukocytes. Neutrophils are part of the first line of defense and have multiple antimicrobial strategies allowing them to attack pathogenic biofilms. Objective/design: In this paper, interaction modes of neutrophils with biofilms are reviewed. Antimicrobial strategies of neutrophils and the counteractions of the biofilm communities, with special attention to oral biofilms, are presented. Moreover, possible adverse effects of neutrophil activity and their biofilm-promoting side effects are discussed. Results/conclusion: Biofilms are partially, but not entirely, protected against neutrophil assault, which include the processes of phagocytosis, degranulation, and formation of neutrophil extracellular traps. However, virulence factors of microorganisms, microbial composition, and properties of the extracellular matrix determine whether a biofilm and subsequent microbial spread can be controlled by neutrophils and other host defense factors. Besides, neutrophils may inadvertently contribute to the physical and ecological stability of biofilms by promoting selection of more resistant strains. Moreover, neutrophil enzymes can degrade collagen and other proteins and, as a result, cause harm to the host tissues. These parameters could be crucial factors in the onset of periodontal inflammation and the subsequent tissue breakdown.

  7. Drug resistance of bacterial dental biofilm and the potential use of natural compounds as alternative for prevention and treatment.

    Science.gov (United States)

    Kouidhi, Bochra; Al Qurashi, Yasir Mohammed A; Chaieb, Kamel

    2015-03-01

    Oral diseases, such as dental caries and periodontal disease are directly linked with the ability of bacteria to form biofilm. The development of dental caries involves acidogenic and aciduric Gram-positive bacteria colonizing the supragingival biofilm (Streptococcus, Lactobacillus and Actinomycetes). Periodontal diseases have been linked to anaerobic Gram-negative bacteria forming a subgingival plaque (Porphyromonas gingivalis, Actinobacillus, Prevotella and Fusobacterium). Cells embedded in biofilm are up to 1000-fold more resistant to antibiotics compared to their planctonic ones. Several mechanisms have been proposed to explain biofilms drug resistance. Given the increased bacterial resistance to antibiotics currently used in dentistry, a great importance is given to natural compounds for the prevention of oral bacterial growth, adhesion and colonization. Over the past decade, interest in drugs derived from medicinal plants has markedly increased. It has been well documented that medicinal plants and natural compounds confer considerable antibacterial activity against various microorganisms including cariogenic and periodontal pathogens. This paper provides a review of the literature focusing on the studies on (i) biofilm in the oral cavity, (ii) drug resistance of bacterial biofilm and (iii) the potential use of plant extracts, essential oils and natural compounds as biofilm preventive agents in dentistry, involving their origin and their mechanism of biofilm inhibition.

  8. Biofilm formation by multidrug resistant Escherichia coli ST131 is dependent on type 1 fimbriae and assay conditions.

    Science.gov (United States)

    Sarkar, Sohinee; Vagenas, Dimitrios; Schembri, Mark A; Totsika, Makrina

    2016-04-01

    Escherichia coli sequence type 131 (ST131) has emerged as a pandemic lineage of important multidrug resistant pathogens worldwide. Despite many studies examining the epidemiology of ST131, only a few studies to date have investigated the capacity of ST131 strains to form biofilms. Some of these studies have reported contrasting findings, with no specific ST131 biofilm-promoting factors identified. Here, we examined a diverse collection of ST131 isolates for in vitro biofilm formation in different media and assay conditions, including urine from healthy adult women. We found significant differences among strains and assay conditions, which offers an explanation for the contrasting findings reported by previous studies using a single condition. Importantly, we showed that expression of type 1 fimbriae is a critical determinant for biofilm formation by ST131 strains and that inhibition of the FimH adhesin significantly reduces biofilm formation. We also offer direct genetic evidence for the contribution of type 1 fimbriae in biofilm formation by the reference ST131 strain EC958, a representative of the clinically dominant H30-Rx ST131 subgroup. This is the first study of ST131 biofilm formation in biologically relevant conditions and paves the way for the application of FimH inhibitors in treating drug resistant ST131 biofilm infections.

  9. Roles of oral bacteria in cardiovascular diseases--from molecular mechanisms to clinical cases: Treatment of periodontal disease regarded as biofilm infection: systemic administration of azithromycin.

    Science.gov (United States)

    Wang, Pao-Li

    2010-01-01

    Periodontal disease as a biofilm infectious disease is considered. Periodontal disease-associated bacteria formed biofilm in periodontal pockets or on the surface of cementum. Planktonic bacteria from biofilm invade into periodontal tissues and lead to inflammation and destruction of tissues directly and indirectly by eliciting the host defense mechanism. Supragingival dental plaques (biofilm) are easily removed by professional mechanical tooth cleaning, while subgingival dental plaques and bacteria invading into periodontal tissues are difficult to remove. Therefore, the development of a method for periodontal disease based on the concept that regards periodontal disease as a biofilm infectious disease is needed. Hereby, I report the effect of antibiotics on an in vitro biofilm model of periodontal disease and the systemic administration of azithromycin for early-onset (aggressive) periodontitis like a treatment resistant periodontitis.

  10. Mycobacterium biofilms: factors involved in development, dispersal, and therapeutic strategies against biofilm-relevant pathogens.

    Science.gov (United States)

    Xiang, Xiaohong; Deng, Wanyan; Liu, Minqiang; Xie, Jianping

    2014-01-01

    Many bacteria can develop biofilm (BF), a multicellular structure largely combining bacteria and their extracellular polymeric substances (EPS). The formation of biofilm results in an alternative existence in which microbes ensure their survival in adverse environments. Biofilm-relevant infections are more persistent, resistant to most antibiotics, and more recalcitrant to host immunity. Mycobacterium tuberculosis, the causative agent of tuberculosis, can develop biofilm, though whether M. tuberculosis can form biofilm within tuberculosis patients has yet to be determined. Here, we summarize the factors involved in the development and dispersal of mycobacterial biofilms, as well as underlying regulatory factors and inhibitors against biofilm to deepen our understanding of their development and to elucidate potential novel modes of action for future antibiotics. Key factors in biofilm formation identified as drug targets represent a novel and promising avenue for developing better antibiotics.

  11. Influence of biofilm thickness on micropollutants removal in nitrifying MBBRs

    DEFF Research Database (Denmark)

    Torresi, Elena; Andersen, Henrik Rasmus; Smets, Barth F.;

    The removal of pharmaceuticals was investigated in nitrifying Moving Bed Biofilm Reactors (MBBRs) containing carriers with different biofilm thicknesses. The biofilm with the thinnest thickness was found to have the highest nitrification and biotransformation rate for some key pharmaceuticals...

  12. Novel metabolic activity indicator in Streptococcus mutans biofilms

    NARCIS (Netherlands)

    Deng, D.M.; Hoogenkamp, M.A.; ten Cate, J.M.; Crielaard, W.

    2009-01-01

    Antimicrobial resistance of micro-organisms in biofilms requires novel strategies to evaluate the efficacy of caries preventive agents in actual biofilms. Hence we investigated fluorescence intensity (FI) in Streptococcus mutans biofilms constitutively expressing green fluorescent protein (GFP). Upo

  13. Inhibition of Biofilm Formation Using Novel Nanostructured Surfaces Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Biofilms are ubiquitous in the environment. Few surfaces resist biofilm formation, most promote it. Biofilm formation poses problems in water systems as they can...

  14. The composition and compression of biofilms developed on ultrafiltration membranes determine hydraulic biofilm resistance.

    Science.gov (United States)

    Derlon, Nicolas; Grütter, Alexander; Brandenberger, Fabienne; Sutter, Anja; Kuhlicke, Ute; Neu, Thomas R; Morgenroth, Eberhard

    2016-10-01

    This study aimed at identifying how to improve the level of permeate flux stabilisation during gravity-driven membrane filtration without control of biofilm formation. The focus was therefore on understanding (i) how the different fractions of the biofilms (inorganics particles, bacterial cells, EPS matrix) influence its hydraulic resistance and (ii) how the compression of biofilms impacts its hydraulic resistance, i.e., can water head be increased to increase the level of permeate flux stabilisation. Biofilms were developed on ultrafiltration membranes at 88 and 284 cm water heads with dead-end filtration for around 50 days. A larger water head resulted in a smaller biofilm permeability (150 and 50 L m(-2) h(-1) bar(-1) for biofilms grown at 88 cm and 284 cm water head, respectively). Biofilms were mainly composed of EPS (>90% in volume). The comparison of the hydraulic resistances of biofilms to model fouling layers indicated that most of the hydraulic resistance is due to the EPS matrix. The compressibility of the biofilm was also evaluated by subjecting the biofilms to short-term (few minutes) and long-term variations of transmembrane pressures (TMP). A sudden change of TMP resulted in an instantaneous and reversible change of biofilm hydraulic resistance. A long-term change of TMP induced a slow change in the biofilm hydraulic resistance. Our results demonstrate that the response of biofilms to a TMP change has two components: an immediate variation of resistance (due to compression/relaxation) and a long-term response (linked to biofilm adaptation/growth). Our results provide relevant information about the relationship between the operating conditions in terms of TMP, the biofilm structure and composition and the resulting biofilm hydraulic resistance. These findings have practical implications for a broad range of membrane systems.

  15. How Staphylococcus aureus biofilms develop their characteristic structure

    OpenAIRE

    Periasamy, Saravanan; Joo, Hwang-Soo; Anthony C. Duong; Bach, Thanh-Huy L.; Tan, Vee Y.; Chatterjee, Som S; Cheung, Gordon Y. C.; Otto, Michael

    2012-01-01

    Biofilms cause significant problems in the environment and during the treatment of infections. However, the molecular mechanisms underlying biofilm formation are poorly understood. There is a particular lack of knowledge about biofilm maturation processes, such as biofilm structuring and detachment, which are deemed crucial for the maintenance of biofilm viability and the dissemination of cells from a biofilm. Here, we identify the phenol-soluble modulin (PSM) surfactant peptides as key biofi...

  16. Innovative Strategies to Overcome Biofilm Resistance

    Directory of Open Access Journals (Sweden)

    Aleksandra Taraszkiewicz

    2013-01-01

    Full Text Available We review the recent literature concerning the efficiency of antimicrobial photodynamic inactivation toward various microbial species in planktonic and biofilm cultures. The review is mainly focused on biofilm-growing microrganisms because this form of growth poses a threat to chronically infected or immunocompromised patients and is difficult to eradicate from medical devices. We discuss the biofilm formation process and mechanisms of its increased resistance to various antimicrobials. We present, based on data in the literature, strategies for overcoming the problem of biofilm resistance. Factors that have potential for use in increasing the efficiency of the killing of biofilm-forming bacteria include plant extracts, enzymes that disturb the biofilm structure, and other nonenzymatic molecules. We propose combining antimicrobial photodynamic therapy with various antimicrobial and antibiofilm approaches to obtain a synergistic effect to permit efficient microbial growth control at low photosensitizer doses.

  17. Silver-Palladium Surfaces Inhibit Biofilm Formation

    DEFF Research Database (Denmark)

    Chiang, Wen-Chi; Schroll, Casper; Hilbert, Lisbeth Rischel

    2009-01-01

    Undesired biofilm formation is a major concern in many areas. In the present study, we investigated biofilm-inhibiting properties of a silver-palladium surface that kills bacteria by generating microelectric fields and electrochemical redox processes. For evaluation of the biofilm inhibition...... efficacy and study of the biofilm inhibition mechanism, the silver-sensitive Escherichia coli J53 and the silver-resistant E. coli J53[pMG101] strains were used as model organisms, and batch and flow chamber setups were used as model systems. In the case of the silver-sensitive strain, the silver......-palladium surfaces killed the bacteria and prevented biofilm formation under conditions of low or high bacterial load. In the case of the silver-resistant strain, the silver-palladium surfaces killed surface-associated bacteria and prevented biofilm formation under conditions of low bacterial load, whereas under...

  18. Mucosal biofilm detection in chronic otitis media

    DEFF Research Database (Denmark)

    Wessman, Marcus; Bjarnsholt, Thomas; Eickhardt-Sørensen, Steffen Robert

    2015-01-01

    The objectives of this study were to examine middle ear biopsies from Greenlandic patients with chronic otitis media (COM) for the presence of mucosal biofilms and the bacteria within the biofilms. Thirty-five middle ear biopsies were obtained from 32 Greenlandic COM patients admitted to ear...... of the patients served as controls. PNA-FISH showed morphological signs of biofilms in 15 out of 35 (43 %) middle ear biopsies. In the control skin biopsies, there were signs of biofilms in eight out of 23 biopsies (30 %), probably representing skin flora. PCR and 16s sequencing detected bacteria in seven out...... of 20 (35 %) usable middle ear biopsies, and in two out of ten (20 %) usable control samples. There was no association between biofilm findings and PCR and 16s sequencing. Staphylococci were the most common bacteria in bacterial culture. We found evidence of bacterial biofilms in 43 % of middle ear...

  19. [Biofilms and their significance in medical microbiology].

    Science.gov (United States)

    Cernohorská, L; Votava, M

    2002-11-01

    Microorganisms are able to adhere to various surfaces and to form there a three-dimensional structure known as biofilm. In biofilms, microbial cells show characteristics and behaviours different from those of plankton cells. Intercellular signalizations of the quorum-sensing type regulate interaction between members of the biofilm. Bacteria embedded in the biofilm can escape and form well known planktonic forms, that are obviously only a part of the bacterial life cycle. Bacteria adhere also to medically important surfaces such as catheters, either urinary or intravenous ones, artificial heart valves, orthopedic implants and so on and contribute to device-related infections like cystitis, catheter-related sepsis, endocarditis etc. Once a biofilm has been established on a surface, the bacteria harboured inside are less exposed to the host's immune response and less susceptible to antibiotics. As an important cause of nosocomial infections the biofilm must remain in the centre of the microbiologist's attention.

  20. Microbial biofilms: biosurfactants as antibiofilm agents.

    Science.gov (United States)

    Banat, Ibrahim M; De Rienzo, Mayri A Díaz; Quinn, Gerry A

    2014-12-01

    Current microbial inhibition strategies based on planktonic bacterial physiology have been known to have limited efficacy on the growth of biofilm communities. This problem can be exacerbated by the emergence of increasingly resistant clinical strains. All aspects of biofilm measurement, monitoring, dispersal, control, and inhibition are becoming issues of increasing importance. Biosurfactants have merited renewed interest in both clinical and hygienic sectors due to their potential to disperse microbial biofilms in addition to many other advantages. The dispersal properties of biosurfactants have been shown to rival those of conventional inhibitory agents against bacterial and yeast biofilms. This makes them suitable candidates for use in new generations of microbial dispersal agents and for use as adjuvants for existing microbial suppression or eradication strategies. In this review, we explore aspects of biofilm characteristics and examine the contribution of biologically derived surface-active agents (biosurfactants) to the disruption or inhibition of microbial biofilms.

  1. Artificial biofilms establish the role of matrix interactions in staphylococcal biofilm assembly and disassembly.

    Science.gov (United States)

    Stewart, Elizabeth J; Ganesan, Mahesh; Younger, John G; Solomon, Michael J

    2015-08-14

    We demonstrate that the microstructural and mechanical properties of bacterial biofilms can be created through colloidal self-assembly of cells and polymers, and thereby link the complex material properties of biofilms to well understood colloidal and polymeric behaviors. This finding is applied to soften and disassemble staphylococcal biofilms through pH changes. Bacterial biofilms are viscoelastic, structured communities of cells encapsulated in an extracellular polymeric substance (EPS) comprised of polysaccharides, proteins, and DNA. Although the identity and abundance of EPS macromolecules are known, how these matrix materials interact with themselves and bacterial cells to generate biofilm morphology and mechanics is not understood. Here, we find that the colloidal self-assembly of Staphylococcus epidermidis RP62A cells and polysaccharides into viscoelastic biofilms is driven by thermodynamic phase instability of EPS. pH conditions that induce phase instability of chitosan produce artificial S. epidermidis biofilms whose mechanics match natural S. epidermidis biofilms. Furthermore, pH-induced solubilization of the matrix triggers disassembly in both artificial and natural S. epidermidis biofilms. This pH-induced disassembly occurs in biofilms formed by five additional staphylococcal strains, including three clinical isolates. Our findings suggest that colloidal self-assembly of cells and matrix polymers produces biofilm viscoelasticity and that biofilm control strategies can exploit this mechanism.

  2. Probing Electrode Heterogeneity Using Fourier-Transformed Alternating Current Voltammetry: Application to a Dual-Electrode Configuration.

    Science.gov (United States)

    Tan, Sze-Yin; Unwin, Patrick R; Macpherson, Julie V; Zhang, Jie; Bond, Alan M

    2017-03-07

    Quantitative studies of electron transfer processes at electrode/electrolyte interfaces, originally developed for homogeneous liquid mercury or metallic electrodes, are difficult to adapt to the spatially heterogeneous nanostructured electrode materials that are now commonly used in modern electrochemistry. In this study, the impact of surface heterogeneity on Fourier-transformed alternating current voltammetry (FTACV) has been investigated theoretically under the simplest possible conditions where no overlap of diffusion layers occurs and where numerical simulations based on a 1D diffusion model are sufficient to describe the mass transport problem. Experimental data that meet these requirements can be obtained with the aqueous [Ru(NH3)6](3+/2+) redox process at a dual-electrode system comprised of electrically coupled but well-separated glassy carbon (GC) and boron-doped diamond (BDD) electrodes. Simulated and experimental FTACV data obtained with this electrode configuration, and where distinctly different heterogeneous charge transfer rate constants (k(0) values) apply at the individual GC and BDD electrode surfaces, are in excellent agreement. Principally, because of the far greater dependence of the AC current magnitude on k(0), it is straightforward with the FTACV method to resolve electrochemical heterogeneities that are ∼1-2 orders of magnitude apart, as applies in the [Ru(NH3)6](3+/2+) dual-electrode configuration experiments, without prior knowledge of the individual kinetic parameters (k(0)1 and k(0)2) or the electrode size ratio (θ1:θ2). In direct current voltammetry, a difference in k(0) of >3 orders of magnitude is required to make this distinction.

  3. Penetration of erythromycin through Staphylococcus epidermidis biofilm

    Institute of Scientific and Technical Information of China (English)

    LIN Mao-hu; HE Lei; GAO Jie; LIU Yun-xi; SUO Ji-jiang; XING Yu-bin; JIA Ning

    2013-01-01

    Background The catheter related infection caused by Staphylococcus epiderrnidis biofilm is increasing and difficult to treat by antimicrobial chemotherapy.The properties of biofilms that give rise to antibiotic resistance are only partially understood.This study aimed to elucidate the penetration of erythromycin through Staphylococcus epidermidis biofilm.Methods The penetration ratio of erythromycin through Staphylococcus epidermidis biofilms of 1457,1457-msrA,and wild isolate S68 was detected by biofilm penetration model at different time points according to the standard regression curve.The RNNDNA ratio and the cell density within the biofilms were observed by confocal laser microscope and transmission electromicroscope,respectively.Results The penetration ratios of erythromycin through the biofilms of 1457,1457-msrA,and S68 after cultivation for 36 hours were 0.93,0.55 and 0.4,respectively.The erythromycin penetration ratio through 1457 biofilm (0.58 after 8 hours)was higher than that through the other two (0.499 and 0.31 after 24 hours).Lower growth rate of the cells in biofilm was shown,with reduction of RNA/DNA proportion observed by confocal laser microscope through acridine orange stain.Compared with the control group observed by transmission electrmicroscope,the cell density of biofilm air face was lower than that of agar face,with more cell debris.Conclusions Erythromycin could penetrate to the Staphylococcus epidermidis biofilm,but could not kill the cells thoroughly.The lower growth rate of the cells within biofilm could help decreasing the erythromycin susceptibility.

  4. Red and Green Fluorescence from Oral Biofilms

    Science.gov (United States)

    Hoogenkamp, Michel A.; Krom, Bastiaan P.; Janus, Marleen M.; ten Cate, Jacob M.; de Soet, Johannes J.; Crielaard, Wim; van der Veen, Monique H.

    2016-01-01

    Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries. PMID:27997567

  5. Polymicrobial biofilms by diabetic foot clinical isolates.

    Science.gov (United States)

    Mottola, Carla; Mendes, João J; Cristino, José Melo; Cavaco-Silva, Patrícia; Tavares, Luís; Oliveira, Manuela

    2016-01-01

    Diabetes mellitus is a major chronic disease that continues to increase significantly. One of the most important and costly complications of diabetes is foot ulceration that may be colonized by pathogenic and antimicrobial resistant bacteria, which may express several virulence factors that could impair treatment success. These bacterial communities can be organized in polymicrobial biofilms, which may be responsible for diabetic foot ulcer (DFU) chronicity. We evaluated the influence of polymicrobial communities in the ability of DFU isolates to produce biofilm, using a microtiter plate assay and a multiplex fluorescent in situ hybridization, at three time points (24, 48, 72 h), after evaluating biofilm formation by 95 DFU isolates belonging to several bacterial genera (Staphylococcus, Corynebacterium, Enterococcus, Pseudomonas and Acinetobacter). All isolates were biofilm-positive at 24 h, and the amount of biofilm produced increased with incubation time. Pseudomonas presented the higher biofilm production, followed by Corynebacterium, Acinetobacter, Staphylococcus and Enterococcus. Significant differences were found in biofilm formation between the three time points. Polymicrobial communities produced higher biofilm values than individual species. Pseudomonas + Enterococcus, Acinetobacter + Staphylococcus and Corynebacterium + Staphylococcus produced higher biofilm than the ones formed by E. faecalis + Staphylococcus and E. faecalis + Corynebacterium. Synergy between bacteria present in dual or multispecies biofilms has been described, and this work represents the first report on time course of biofilm formation by polymicrobial communities from DFUs including several species. The biological behavior of different bacterial species in polymicrobial biofilms has important clinical implications for the successful treatment of these infections.

  6. Antibiotic tolerance and resistance in biofilms

    DEFF Research Database (Denmark)

    Ciofu, Oana; Tolker-Nielsen, Tim

    2010-01-01

    One of the most important features of microbial biofilms is their tolerance to antimicrobial agents and components of the host immune system. The difficulty of treating biofilm infections with antibiotics is a major clinical problem. Although antibiotics may decrease the number of bacteria...... in biofilms, they will not completely eradicate the bacteria in vivo which may have important clinical consequences in form of relapses of the infection....

  7. Development and Use of a Cyclic Voltammetry Simulator to Introduce Undergraduate Students to Electrochemical Simulations

    Science.gov (United States)

    Brown, Jay H.

    2015-01-01

    Cyclic voltammetry (CV) is a popular technique for the study of electrochemical mechanisms because the method can provide useful information on the redox couple. The technique involves the application of a potential ramp on an unstirred solution while the current is monitored, and then the ramp is reversed for a return sweep. CV is sometimes…

  8. Theory of linear sweep voltammetry with diffuse charge: Unsupported electrolytes, thin films, and leaky membranes

    Science.gov (United States)

    Yan, David; Bazant, Martin Z.; Biesheuvel, P. M.; Pugh, Mary C.; Dawson, Francis P.

    2017-03-01

    Linear sweep and cyclic voltammetry techniques are important tools for electrochemists and have a variety of applications in engineering. Voltammetry has classically been treated with the Randles-Sevcik equation, which assumes an electroneutral supported electrolyte. In this paper, we provide a comprehensive mathematical theory of voltammetry in electrochemical cells with unsupported electrolytes and for other situations where diffuse charge effects play a role, and present analytical and simulated solutions of the time-dependent Poisson-Nernst-Planck equations with generalized Frumkin-Butler-Volmer boundary conditions for a 1:1 electrolyte and a simple reaction. Using these solutions, we construct theoretical and simulated current-voltage curves for liquid and solid thin films, membranes with fixed background charge, and cells with blocking electrodes. The full range of dimensionless parameters is considered, including the dimensionless Debye screening length (scaled to the electrode separation), Damkohler number (ratio of characteristic diffusion and reaction times), and dimensionless sweep rate (scaled to the thermal voltage per diffusion time). The analysis focuses on the coupling of Faradaic reactions and diffuse charge dynamics, although capacitive charging of the electrical double layers is also studied, for early time transients at reactive electrodes and for nonreactive blocking electrodes. Our work highlights cases where diffuse charge effects are important in the context of voltammetry, and illustrates which regimes can be approximated using simple analytical expressions and which require more careful consideration.

  9. The Rise of Voltammetry: From Polarography to the Scanning Electrochemical Microscope

    Science.gov (United States)

    Bard, Allen J.

    2007-01-01

    The drooping mercury electrode (DME) was previously used to carry out electrochemical experiments but invention of polarography technique changed this. Voltammetry with DME was given the term polarography and are used in measurement of current as a function of potential at small electrodes.

  10. Lead-Testing Service to Elementary and Secondary Schools Using Anodic Stripping Voltammetry

    Science.gov (United States)

    Goebel, Amanda; Vos, Tracy; Louwagie, Anne; Lundbohm, Laura; Brown, Jay H.

    2004-01-01

    The undergraduate chemistry club of the Southwest Minnesota State University offers assistance in lead-testing through the anodic stripping voltammetry (ASV) technique to elementary and secondary schools. Emphasis is given to this community service activity, which has increased club membership, and promoted discussion of water quality problems in…

  11. Adsorptive stripping voltammetry of antibiotics rifamycin SV and rifampicin at renewable pencil electrodes.

    Science.gov (United States)

    Kawde, Abdel-Nasser; Temerk, Yassein; Farhan, Nasser

    2014-01-01

    Adsorptive stripping voltammetry of antibiotics of rifamycin SV (RSV) and rifampicin (RIF) was investigated by cyclic voltammetry and differential pulse voltammetry using a renewable pencil graphite electrode (PGE). The nature of the oxidation process of RSV and RIF taking place at the PGE was characterized. The results show that the determination of highly sensitive oxidation peak current is the basis of a simple, accurate and rapid method for quantification of RSV and RIF in bulk forms, pharmaceutical formulations and biological fluids by differential pulse adsorptive stripping voltammetry (DPASV). Factors influencing the trace measurement of RSV and RIF at PGE are assessed. The limits of detection for the determination of RSV and RIF in bulk forms are 6.0 × 10(-8) mol/L and 1.3 × 10(-8) mol/L, respectively. Moreover, the proposed procedure was successfully applied to assay both RSV and RIF in pharmaceutical formulations and in biological fluids. The capability of the proposed procedure for simultaneous assay of antibiotics RSV-isoniazid and RIF-isoniazid was achieved. The statistical analysis and calibration curve data for trace determination of RSV and RIF are reported.

  12. Characterization of an electrochemical mercury sensor using alternating current, cyclic, square wave and differential pulse voltammetry.

    Science.gov (United States)

    Guerreiro, Gabriela V; Zaitouna, Anita J; Lai, Rebecca Y

    2014-01-31

    Here we report the characterization of an electrochemical mercury (Hg(2+)) sensor constructed with a methylene blue (MB)-modified and thymine-containing linear DNA probe. Similar to the linear probe electrochemical DNA sensor, the resultant sensor behaved as a "signal-off" sensor in alternating current voltammetry and cyclic voltammetry. However, depending on the applied frequency or pulse width, the sensor can behave as either a "signal-off" or "signal-on" sensor in square wave voltammetry (SWV) and differential pulse voltammetry (DPV). In SWV, the sensor showed "signal-on" behavior at low frequencies and "signal-off" behavior at high frequencies. In DPV, the sensor showed "signal-off" behavior at short pulse widths and "signal-on" behavior at long pulse widths. Independent of the sensor interrogation technique, the limit of detection was found to be 10nM, with a linear dynamic range between 10nM and 500nM. In addition, the sensor responded to Hg(2+) rather rapidly; majority of the signal change occurred in <20min. Overall, the sensor retains all the characteristics of this class of sensors; it is reagentless, reusable, sensitive, specific and selective. This study also highlights the feasibility of using a MB-modified probe for real-time sensing of Hg(2+), which has not been previously reported. More importantly, the observed "switching" behavior in SWV and DPV is potentially generalizable and should be applicable to most sensors in this class of dynamics-based electrochemical biosensors.

  13. Determination of bosentan in pharmaceutical preparations by linear sweep, square wave and differential pulse voltammetry methods.

    Science.gov (United States)

    Atila, Alptug; Yilmaz, Bilal

    2015-01-01

    In this study, simple, fast and reliable cyclic voltammetry (CV), linear sweep voltammetry (LSV), square wave voltammetry (SWV) and differential pulse voltammetry (DPV) methods were developed and validated for determination of bosentan in pharmaceutical preparations. The proposed methods were based on electrochemical oxidation of bosentan at platinum electrode in acetonitrile solution containing 0.1 M TBACIO4. The well-defined oxidation peak was observed at 1.21 V. The calibration curves were linear for bosentan at the concentration range of 5-40 µg/mL for LSV and 5-35 µg/mL for SWV and DPV methods, respectively. Intra- and inter-day precision values for bosentan were less than 4.92, and accuracy (relative error) was better than 6.29%. The mean recovery of bosentan was 100.7% for pharmaceutical preparations. No interference was found from two tablet excipients at the selected assay conditions. Developed methods in this study are accurate, precise and can be easily applied to Tracleer and Diamond tablets as pharmaceutical preparation.

  14. Kinetic Diversity of Striatal Dopamine: Evidence from a Novel Protocol for Voltammetry.

    Science.gov (United States)

    Walters, Seth H; Robbins, Elaine M; Michael, Adrian C

    2016-05-18

    In vivo voltammetry reveals substantial diversity of dopamine kinetics in the rat striatum. To substantiate this kinetic diversity, we evaluate the temporal distortion of dopamine measurements arising from the diffusion-limited adsorption of dopamine to voltammetric microelectrodes. We validate two mathematical procedures for correcting adsorptive distortion, both of which substantiate that dopamine's apparent kinetic diversity is not an adsorption artifact.

  15. THE TRANSFER OF TMA+ ION AT THE WATERN NITROBENZENE INTERFACE USING SEMI-DIFFERENTIAL CYCLIC VOLTAMMETRY

    Institute of Scientific and Technical Information of China (English)

    范瑞溪; 王小平

    1990-01-01

    This paper describes the investigation of the electrochemical phenomena of TMA+ transfer at the w/nb interface using semi-differential cyclic voltammetry. The expressions for the peak potential, half-wave width and peak current are derived. All the experimental results are in good agreement with the theoretical ones.

  16. Adsorptive Stripping Voltammetry of Environmental Indicators: Determination of Zinc in Algae

    Science.gov (United States)

    Collado-Sanchez, C.; Hernandez-Brito, J. J.; Perez-Pena, J.; Torres-Padron, M. E.; Gelado-Caballero, M. D.

    2005-01-01

    A method for sample preparation and for the determination of average zinc content in algae using adsorptive stripping voltammetry are described. The students gain important didactic advantages through metal determination in environmental matrices, which include carrying out clean protocols for sampling and handling, and digesting samples using…

  17. Cyclic Voltammetry Simulations with DigiSim Software: An Upper-Level Undergraduate Experiment

    Science.gov (United States)

    Messersmith, Stephania J.

    2014-01-01

    An upper-division undergraduate chemistry experiment is described which utilizes DigiSim software to simulate cyclic voltammetry (CV). Four mechanisms were studied: a reversible electron transfer with no subsequent or proceeding chemical reactions, a reversible electron transfer followed by a reversible chemical reaction, a reversible chemical…

  18. Differential Cyclic Voltammetry - a Novel Technique for Selective and Simultaneous Detection using Redox Cycling Based Sensors

    NARCIS (Netherlands)

    Odijk, M.; Wiedemair, J.; Megen, M.J.J; Olthuis, W.; Berg, van den A.

    2010-01-01

    Redox cycling (RC) is an effect that is used to amplify electrochemical signals. However, traditional techniques such as cyclic voltammetry (CV) do not provide clear insight for a mixture of multiple redox couples while RC is applied. Thus, we have developed a new measurement technique which deliver

  19. Characterization of Fe implanted yttria-stabilized zirconia by cyclic voltammetry

    NARCIS (Netherlands)

    Hassel, van B.A.; Burggraaf, A.J.

    1992-01-01

    The technique of cyclic voltammetry has been applied to study reduction and oxidation phenomena which are observed at low oxygen partial pressures during steady state current-overpotential measurements of the Au, O2(g)/Fe implanted yttria-stabilized zirconia interface. The redox potential (EO) of th

  20. Polymyxin-coated Au and carbon nanotube electrodes for stable [NiFe]-hydrogenase film voltammetry.

    NARCIS (Netherlands)

    Hoeben, F.J.M.; Heller, I.; Albracht, S.P.J.; Dekker, C.; Lemay, S.G.; Heering, H.A.

    2008-01-01

    We report on the use of polymyxin (PM), a cyclic cationic lipodecapeptide, as an electrode modifier for studying protein film voltammetry (PFV) on Au and single-walled carbon nanotube (SWNT) electrodes. Pretreating the electrodes with PM allows for the subsequent immobilization of an active submonol

  1. Aspartate inhibits Staphylococcus aureus biofilm formation.

    Science.gov (United States)

    Yang, Hang; Wang, Mengyue; Yu, Junping; Wei, Hongping

    2015-04-01

    Biofilm formation renders Staphylococcus aureus highly resistant to conventional antibiotics and host defenses. Four D-amino acids (D-Leu, D-Met, D-Trp and D-Tyr) have been reported to be able to inhibit biofilm formation and disassemble established S. aureus biofilms. We report here for the first time that both D- and L-isoforms of aspartate (Asp) inhibited S. aureus biofilm formation on tissue culture plates. Similar biofilm inhibition effects were also observed against other staphylococcal strains, including S. saprophyticus, S. equorum, S. chromogenes and S. haemolyticus. It was found that Asp at high concentrations (>10 mM) inhibited the growth of planktonic N315 cells, but at subinhibitory concentrations decreased the cellular metabolic activity without influencing cell growth. The decreased cellular metabolic activity might be the reason for the production of less protein and DNA in the matrix of the biofilms formed in the presence of Asp. However, varied inhibition efficacies of Asp were observed for biofilms formed by clinical staphylococcal isolates. There might be mechanisms other than decreasing the metabolic activity, e.g. the biofilm phenotypes, affecting biofilm formation in the presence of Asp.

  2. Focus on the physics of biofilms

    Science.gov (United States)

    Lecuyer, Sigolene; Stocker, Roman; Rusconi, Roberto

    2015-03-01

    Bacteria are the smallest and most abundant form of life. They have traditionally been considered as primarily planktonic organisms, swimming or floating in a liquid medium, and this view has shaped many of the approaches to microbial processes, including for example the design of most antibiotics. However, over the last few decades it has become clear that many bacteria often adopt a sessile, surface-associated lifestyle, forming complex multicellular communities called biofilms. Bacterial biofilms are found in a vast range of environments and have major consequences on human health and industrial processes, from biofouling of surfaces to the spread of diseases. Although the study of biofilms has been biologists’ territory for a long time, a multitude of phenomena in the formation and development of biofilms hinges on physical processes. We are pleased to present a collection of research papers that discuss some of the latest developments in many of the areas to which physicists can contribute a deeper understanding of biofilms, both experimentally and theoretically. The topics covered range from the influence of physical environmental parameters on cell attachment and subsequent biofilm growth, to the use of local probes and imaging techniques to investigate biofilm structure, to the development of biofilms in complex environments and the modeling of colony morphogenesis. The results presented contribute to addressing some of the major challenges in microbiology today, including the prevention of surface contamination, the optimization of biofilm disruption methods and the effectiveness of antibiotic treatments.

  3. In situ rheology of yeast biofilms.

    Science.gov (United States)

    Brugnoni, Lorena I; Tarifa, María C; Lozano, Jorge E; Genovese, Diego

    2014-01-01

    The aim of the present work was to investigate the in situ rheological behavior of yeast biofilms growing on stainless steel under static and turbulent flow. The species used (Rhodototula mucilaginosa, Candida krusei, Candida kefyr and Candida tropicalis) were isolated from a clarified apple juice industry. The flow conditions impacted biofilm composition over time, with a predominance of C. krusei under static and turbulent flow. Likewise, structural variations occurred, with a tighter appearance under dynamic flow. Under turbulent flow there was an increase of 112 μm in biofilm thickness at 11 weeks (p rheology and contribute to a thin body of knowledge about fungal biofilm formation.

  4. Biofilm responses to marine fish farm wastes

    Energy Technology Data Exchange (ETDEWEB)

    Sanz-Lazaro, Carlos, E-mail: carsanz@um.es [Departamento de Ecologia e Hidrologia, Facultad de Biologia, Universidad de Murcia, 30100 Murcia (Spain); Navarrete-Mier, Francisco; Marin, Arnaldo [Departamento de Ecologia e Hidrologia, Facultad de Biologia, Universidad de Murcia, 30100 Murcia (Spain)

    2011-03-15

    The changes in the biofilm community due to organic matter enrichment, eutrophication and metal contamination derived from fish farming were studied. The biofilm biomass, polysaccharide content, trophic niche and element accumulation were quantified along an environmental gradient of fish farm wastes in two seasons. Biofilm structure and trophic diversity was influenced by seasonality as well as by the fish farm waste load. Fish farming enhanced the accumulation of organic carbon, nutrients, selenium and metals by the biofilm community. The accumulation pattern of these elements was similar regardless of the structure and trophic niche of the community. This suggests that the biofilm communities can be considered a reliable tool for assessing dissolved aquaculture wastes. Due to the ubiquity of biofilms and its wide range of consumers, its role as a sink of dissolved wastes may have important implications for the transfer of aquaculture wastes to higher trophic levels in coastal systems. - Research highlights: > Biofilms can act as a trophic pathway of fish farm dissolved wastes. > Biofilms are reliable tools for monitoring fish farm dissolved wastes. > The influence of the fish farm dissolved wastes can be detected 120-350 m from farm. - Under the influence of fish farming biofilm accumulates organic carbon, nutrients, selenium and metals, regardless of the structure and trophic niche of the community.

  5. Microbial biofilms: from ecology to molecular genetics.

    Science.gov (United States)

    Davey, M E; O'toole, G A

    2000-12-01

    Biofilms are complex communities of microorganisms attached to surfaces or associated with interfaces. Despite the focus of modern microbiology research on pure culture, planktonic (free-swimming) bacteria, it is now widely recognized that most bacteria found in natural, clinical, and industrial settings persist in association with surfaces. Furthermore, these microbial communities are often composed of multiple species that interact with each other and their environment. The determination of biofilm architecture, particularly the spatial arrangement of microcolonies (clusters of cells) relative to one another, has profound implications for the function of these complex communities. Numerous new experimental approaches and methodologies have been developed in order to explore metabolic interactions, phylogenetic groupings, and competition among members of the biofilm. To complement this broad view of biofilm ecology, individual organisms have been studied using molecular genetics in order to identify the genes required for biofilm development and to dissect the regulatory pathways that control the plankton-to-biofilm transition. These molecular genetic studies have led to the emergence of the concept of biofilm formation as a novel system for the study of bacterial development. The recent explosion in the field of biofilm research has led to exciting progress in the development of new technologies for studying these communities, advanced our understanding of the ecological significance of surface-attached bacteria, and provided new insights into the molecular genetic basis of biofilm development.

  6. Electroactive biofilms of sulphate reducing bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Cordas, Cristina M.; Guerra, L. Tiago; Xavier, Catarina [Requimte-CQFB, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); Moura, Jose J.G. [Requimte-CQFB, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal)], E-mail: jose.moura@dq.fct.unl.pt

    2008-12-01

    Biofilms formed from a pure strain of Desulfovibrio desulfuricans 27774 on stainless steel and graphite polarised surfaces were studied. The polarisation conditions applied were -0.4 V vs. SCE for different times. A cathodic current related with the biofilms growth was observed with a maximum intensity of -270 mA m{sup -2} that remained stable for several days using graphite electrodes. These sulphate reducing bacteria biofilms present electrocatalytic activity towards hydrogen and oxygen reduction reactions. Electrode polarisation has a selective effect on the catalytic activity. The biofilms were also observed by scanning electronic microscopy revealing the formation of homogeneous films on the surfaces.

  7. The clinical impact of bacterial biofilms

    DEFF Research Database (Denmark)

    Høiby, Niels; Ciofu, Oana; Johansen, Helle Krogh

    2011-01-01

    . Bacterial biofilms are resistant to antibiotics, disinfectant chemicals and to phagocytosis and other components of the innate and adaptive inflammatory defense system of the body. It is known, for example, that persistence of staphylococcal infections related to foreign bodies is due to biofilm formation...... and increased doubling times. These more or less dormant cells are therefore responsible for some of the tolerance to antibiotics. Biofilm growth is associated with an increased level of mutations. Bacteria in biofilms communicate by means of molecules, which activates certain genes responsible for production...

  8. Application of laser scanning microscopy for the analysis of oral biofilm dissolution by different endodontic irrigants

    Directory of Open Access Journals (Sweden)

    Aldo del Carpio-Perochena

    2014-01-01

    Full Text Available Background: Multi-specie biofilms are highly resistant to antimicrobials due to cellular interactions found in them. The purpose of this study was to evaluate, by confocal laser scanning microscopy, the biofilm dissolution effectiveness of different irrigant solutions on biofilms developed on infected dentin in situ. Materials and Methods: A total of 120 bovine dentin specimens infected intraorally (30/group were treated by the following solutions: 2% of chlorhexidine digluconate, 1%, 2.5% and 5.25% of sodium hypochlorite (NaOCl. The solutions were utilized for 5, 15 and 30 min with 2 experimental volumes 500 μL and 1 mL. All the samples were stained using an acridine orange and the biofilm thickness before (control group and after the experiments were evaluated, utilizing a confocal microscope at ×40. The Mann-Whitney U and the nom-parametric Kruskal-Wallis Dunns tests were utilized to determine the influence of the volume and to perform the comparisons among the groups respectively. The significance level was set at P 0.05. The biofilm dissolution treated with 1% NaOCl was directly proportional to the exposure time (P 0.05. Conclusion: The higher exposure times and concentrations of NaOCl were not sufficient to dissolve 100% of the biofilm. However, all NaOCl solutions were more effective than 2% chlorhexidine digluconate to dissolve organic matter.

  9. Acinetobacter baumannii biofilms: variations among strains and correlations with other cell properties.

    Science.gov (United States)

    McQueary, Christin N; Actis, Luis A

    2011-04-01

    Acinetobacter baumannii is an opportunistic pathogen that causes serious infections in humans by colonizing and persisting on surfaces normally found in hospital settings. The capacity of this pathogen to persist in these settings could be due to its ability to form biofilms on inanimate surfaces. This report shows that although the ATCC 19606(T) type strain and 8 different clinical isolates form biofilms, there are significant variations in the cell density and microscopic structures of these cell aggregates, with 3 of the isolates forming pellicles floating on the surface of stagnant broth cultures. PCR indicated that, like ATCC 19606(T), all 8 clinical isolates harbor all the genetic components of the CsuA/BABCDE chaperone-usher pili assembly system, which is needed for biofilm formation on plastic. Pili detection in cells of all strains examined supports the presence and function of a pilus assembly system. However, only one of them produced the putative ATCC 19606(T) CsuA/B pilin subunit protein. Hydrophobicity tests and motility assays also showed significant variations among all tested strains and did not result in direct correlations between the biofilm phenotype and cell properties that could affect biofilm formation on abiotic surfaces. This lack of correlation among these 3 phenotypes may reflect some of the variations already reported with this pathogen, which may pose a challenge in the treatment of the infections this pathogen causes in humans using biofilm formation on abiotic surfaces as a target.

  10. Contribution of the Pseudomonas fluorescens MFE01 Type VI Secretion System to Biofilm Formation

    Science.gov (United States)

    Gallique, Mathias; Decoin, Victorien; Barbey, Corinne; Rosay, Thibaut; Feuilloley, Marc G. J.; Orange, Nicole

    2017-01-01

    Type VI secretion systems (T6SSs) are widespread in Gram-negative bacteria, including Pseudomonas. These macromolecular machineries inject toxins directly into prokaryotic or eukaryotic prey cells. Hcp proteins are structural components of the extracellular part of this machinery. We recently reported that MFE01, an avirulent strain of Pseudomonas fluorescens, possesses at least two hcp genes, hcp1 and hcp2, encoding proteins playing important roles in interbacterial interactions. Indeed, P. fluorescens MFE01 can immobilise and kill diverse bacteria of various origins through the action of the Hcp1 or Hcp2 proteins of the T6SS. We show here that another Hcp protein, Hcp3, is involved in killing prey cells during co-culture on solid medium. Even after the mutation of hcp1, hcp2, or hcp3, MFE01 impaired biofilm formation by MFP05, a P. fluorescens strain isolated from human skin. These mutations did not reduce P. fluorescens MFE01 biofilm formation, but the three Hcp proteins were required for the completion of biofilm maturation. Moreover, a mutant with a disruption of one of the unique core component genes, MFE01ΔtssC, was unable to produce its own biofilm or inhibit MFP05 biofilm formation. Finally, MFE01 did not produce detectable N-acyl-homoserine lactones for quorum sensing, a phenomenon reported for many other P. fluorescens strains. Our results suggest a role for the T6SS in communication between bacterial cells, in this strain, under biofilm conditions. PMID:28114423

  11. In Vitro Assessment of Electric Currents Increasing the Effectiveness of Vancomycin Against Staphylococcus epidermidis Biofilms.

    Science.gov (United States)

    Haddad, Peter A; Mah, Thien-Fah; Mussivand, Tofy

    2016-08-01

    Biofilms are communities of bacteria that can cause infections which are resistant to the immune system and antimicrobial treatments, posing a significant threat for patients with implantable and indwelling medical devices. The purpose of our research was to determine if utilizing specific parameters for electric currents in conjunction with antibiotics could effectively treat a highly resistant biofilm. Our study evaluated the impact of 16 μg/mL of vancomycin with or without 22 or 333 μA of direct electric current (DC) generated by stainless steel electrodes against 24-, 48-, and 72-h-old Staphylococcus epidermidis biofilms formed on titanium coupons. An increase in effectiveness of vancomycin was observed with the combination of 333 μA of electric current against 48-h-old biofilms (P value = 0.01) as well as in combination with 22 μA of electric current against 72-h-old biofilms (P value = 0.04); 333 μA of electric current showed the most significant impact on the effectiveness of vancomycin against S. epidermidis biofilms demonstrating a bioelectric effect previously not observed against this strain of bacteria.

  12. Biofilm formation in Acinetobacter baumannii.

    Science.gov (United States)

    Longo, Francesca; Vuotto, Claudia; Donelli, Gianfranco

    2014-04-01

    Acinetobacter baumannii has received much attention in recent years because of its increasing involvement in a number of severe infections and outbreaks occurring in clinical settings, and presumably related to its ability to survive and persist in hospital environments. The treatment of infections caused by A. baumannii nosocomial strains has become increasingly problematic, due to their intrinsic and/or acquired resistance to multiple classes of antibiotics. Furthermore, the demonstrated ability of nosocomial strains to grow as biofilm is believed to play a significant role in their persistence and antibiotic resistance. This review summarises current knowledge on A. baumannii biofilm formation and its clinical significance, as well as the related genetic determinants and the regulation of this process.

  13. Effects of low arsenic concentration exposure on freshwater fish in the presence of fluvial biofilms.

    Science.gov (United States)

    Tuulaikhuu, Baigal-Amar; Bonet, Berta; Guasch, Helena

    2016-02-15

    Arsenic (As) is a highly toxic element and its carcinogenic effect on living organisms is well known. However, predicting real effects in the environment requires an ecological approach since toxicity is influenced by many environmental and biological factors. The purpose of this paper was to evaluate if environmentally-realistic arsenic exposure causes toxicity to fish. An experiment with four different treatments (control (C), biofilm (B), arsenic (+As) and biofilm with arsenic (B+As)) was conducted and each one included sediment to enhance environmental realism, allowing the testing of the interactive effects of biofilm and arsenic on the toxicity to fish. Average arsenic exposure to Eastern mosquitofish (Gambusia holbrooki) was 40.5 ± 7.5 μg/L for +As treatment and 34.4 ± 1.4 μg/L for B+As treatment for 56 days. Fish were affected directly and indirectly by this low arsenic concentration since exposure did not only affect fish but also the function of periphytic biofilms. Arsenic effects on the superoxide dismutase (SOD) and glutathione reductase (GR) activities in the liver of mosquitofish were ameliorated in the presence of biofilms at the beginning of exposure (day 9). Moreover, fish weight gaining was only affected in the treatment without biofilm. After longer exposure (56 days), effects of exposure were clearly seen. Fish showed a marked increase in the catalase (CAT) activity in the liver but the interactive influence of biofilms was not further observed since the arsenic-affected biofilm had lost its role in water purification. Our results highlight the interest and application of incorporating some of the complexity of natural systems in ecotoxicology and support the use of criterion continuous concentration (CCC) for arsenic lower than 150 μg/L and closer to the water quality criteria to protect aquatic life recommended by the Canadian government which is 5 μg As/L.

  14. Proteus mirabilis biofilm protection against struvite crystal dissolution and its implications in struvite urolithiasis.

    Science.gov (United States)

    McLean, R J; Lawrence, J R; Korber, D R; Caldwell, D E

    1991-10-01

    Proteus mirabilis biofilm formation, struvite (MgNH4PO4.6H2O) crystal formation and dissolution in an artificial urine mixture were monitored using computer-enhanced microscopy (CEM) and a 1 x 3 mm. glass flow cell. Image analysis showed that P. mirabilis biofilm formation did not occur to any extent at macroenvironment flow rates greater than two mL/h (equivalent to a microenvironment flow rate of less than 5 microns./sec). Essentially, cells attached to glass surfaces, grew slowly and divided. Daughter cells were generally released directly into the medium where they could then presumably colonize other regions. Microcolonies formed by the adhesion of aggregates of cells from the medium, and over time grew into biofilms. Struvite crystallization due to urease activity and pH elevation above neutrality, was preceded by the deposition of organic matter on the glass surface, followed by the appearance of a number of tiny (one to two microns.) crystals. Crystals forming within a biofilm at low dilution rates took on a characteristic twinned or "X-shaped" appearance (crystal habit) indicative of a rapid growth rate. Those forming outside the biofilm took on a more tabular appearance reflecting their slower growth. When the macroenvironment flow rate of artificial urine (initial pH 5.8) in the glass flow cell was increased from two mL/h to four mL/h, struvite crystals not associated with biofilms dissolved within five to 10 min. Crystals entrapped within the P. mirabilis biofilm withstood flow rates up to 200 mL/h presumably due to the maintenance of an alkaline Mg-saturated microenvironment within the biofilm. These observations may suggest a mechanism by which struvite calculi can grow in spite of neutral or acidic urine pH and resist mild acidification therapy.

  15. Biofilms bacterianos e infección Bacterial biofilms and infection

    Directory of Open Access Journals (Sweden)

    I. Lasa

    2005-08-01

    describe el papel que juegan los biofilms en infecciones humanas persistentes.In developed countries we tend to think of heart disease and the numerous forms of cancer as the main causes of mortality, but on a global scale infectious diseases come close, or may even be ahead: 14.9 million deaths in 2002 compared to cardiovascular diseases (16.9 million deaths and cancer (7.1 million deaths (WHO report 2004. The infectious agents responsible for human mortality have evolved as medical techniques and hygienic measures have changed. Modern-day acute infectious diseases caused by specialized bacterial pathogens such as diphtheria, tetanus, cholera, plague, which represented the main causes of death at the beginning of XX century, have been effectively controlled with antibiotics and vaccines. In their place, more than half of the infectious diseases that affect mildly immunocompromised patients involve bacterial species that are commensal with the human body; these can produce chronic infections, are resistant to antimicrobial agents and there is no effective vaccine against them. Examples of these infections are the otitis media, native valve endocarditis, chronic urinary infections, bacterial prostatitis, osteomyelitis and all the infections related to medical devices. Direct analysis of the surface of medical devices or of tissues that have been foci of chronic infections shows the presence of large numbers of bacteria surrounded by an exopolysaccharide matrix, which has been named the "biofilm". Inside the biofilm, bacteria grow protected from the action of the antibodies, phagocytic cells and antimicrobial treatments. In this article, we describe the role of bacterial biofilms in human persistent infections.

  16. Antimicrobial and biofilm inhibiting diketopiperazines.

    Science.gov (United States)

    de Carvalho, M P; Abraham, W-R

    2012-01-01

    Diketopiperazines are the smallest cyclic peptides known. 90% of Gram-negative bacteria produce diketopiperazines and they have also been isolated from Gram-positive bacteria, fungi and higher organisms. Biosynthesis of cyclodipeptides can be achieved by dedicated nonribosomal peptide synthetases or by a novel type of synthetases named cyclopeptide synthases. Since the first report in 1924 a large number of bioactive diketopiperazines was discovered spanning activities as antitumor, antiviral, antifungal, antibacterial, antiprion, antihyperglycemic or glycosidase inhibitor agents. As infections are of increasing concern for human health and resistances against existing antibiotics are growing this review focuses on the antimicrobial activities of diketopiperazines. The antibiotic bicyclomycin is a diketopiperazine and structure activity studies revealed the unique nature of this compound which was finally developed for clinical applications. The antimicrobial activities of a number of other diketopiperazines along with structure activity relationships are discussed. Here a special focus is on the activity-toxicity problem of many compounds setting tight limitations to their application as drugs. Not only these classical antimicrobial activities but also proposed action in modulating bacterial communication as a new target to control biofilms will be evaluated. Pathogens organized in biofilms are difficult to eradicate because of the increase of their tolerance for antibiotics for several orders. Diketopiperazines were reported to modulate LuxR-mediated quorum-sensing systems of bacteria, and they are considered to influence cell-cell signaling offering alternative ways of biofilm control by interfering with microbial communication. Concluding the review we will finally discuss the potential of diketopiperazines in the clinic to erase biofilm infections.

  17. 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.

  18. Molecular methods for biofilms

    KAUST Repository

    Ferrera, Isabel

    2014-08-30

    at the same time and to compare bacterial communities among different samples or in a single sample after certain treatments. DGGE, T-RFLP and ARISA share similar steps but require different materials and equipment. The three methods involve (i) sampling of the biofilms; (ii) DNA extraction and quantification; and (iii) PCR using specific primers. Metagenomics: This chapter focuses classical and next-generation metagenomics methods. These are limited to bacterial artificial chromosome (BAC) and Fosmid libraries and Sanger and next-generation 454 sequencing, as these methods are currently the most frequently used in research. The chapter discusses the special handling of deoxyribonucleic acid (DNA) needed to construct BAC and Fosmid libraries from marine water samples. It also briefly addresses the related topics of library archiving, databasing, and screening. The chapter provides a high-level overview of the special handling methods required to prepare DNA for BAC library construction. No special handling is needed for Fosmid library construction.

  19. Fur is a repressor of biofilm formation in Yersinia pestis.

    Directory of Open Access Journals (Sweden)

    Fengjun Sun

    Full Text Available BACKGROUND: Yersinia pestis synthesizes the attached biofilms in the flea proventriculus, which is important for the transmission of this pathogen by fleas. The hmsHFRS operons is responsible for the synthesis of exopolysaccharide (the major component of biofilm matrix, which is activated by the signaling molecule 3', 5'-cyclic diguanylic acid (c-di-GMP synthesized by the only two diguanylate cyclases HmsT, and YPO0449 (located in a putative operonYPO0450-0448. METHODOLOGY/PRINCIPAL FINDINGS: The phenotypic assays indicated that the transcriptional regulator Fur inhibited the Y. pestis biofilm production in vitro and on nematode. Two distinct Fur box-like sequences were predicted within the promoter-proximal region of hmsT, suggesting that hmsT might be a direct Fur target. The subsequent primer extension, LacZ fusion, electrophoretic mobility shift, and DNase I footprinting assays disclosed that Fur specifically bound to the hmsT promoter-proximal region for repressing the hmsT transcription. In contrast, Fur had no regulatory effect on hmsHFRS and YPO0450-0448 at the transcriptional level. The detection of intracellular c-di-GMP levels revealed that Fur inhibited the c-di-GMP production. CONCLUSIONS/SIGNIFICANCE: Y. pestis Fur inhibits the c-di-GMP production through directly repressing the transcription of hmsT, and thus it acts as a repressor of biofilm formation. Since the relevant genetic contents for fur, hmsT, hmsHFRS, and YPO0450-0448 are extremely conserved between Y. pestis and typical Y. pseudotuberculosis, the above regulatory mechanisms can be applied to Y. pseudotuberculosis.

  20. Protein-based biofilm matrices in Staphylococci

    Directory of Open Access Journals (Sweden)

    Pietro eSpeziale

    2014-12-01

    Full Text Available Staphylococcus aureus and Staphylococcus epidermidis are the most important etiological agents of biofilm associated-infections on indwelling medical devices. Biofilm infections may also develop independently of indwelling devices, e.g. in native valve endocarditis, bone tissue and open wounds. After attachment to tissue or indwelling medical devices that have been conditioned with host plasma proteins, staphylococcal biofilms grow and produce a specific environment which provides the conditions for cell-cell interaction and formation of multicellular communities. Bacteria living in biofilms express a variety of macromolecules, including exopolysaccharides, proteins, extracellular eDNA and other polymers. The S. aureus surface protein C and G (SasC and SasG, clumping factor B (ClfB, serine aspartate repeat protein (SdrC, the biofilm-associated protein (Bap and the fibronectin/fibrinogen-binding proteins (FnBPA and FnBPB are individually implicated in biofilm matrix formation. In S. epidermidis, a protein named accumulation-associated protein (Aap contributes to both the primary attachment phase and the establishment of intercellular connections by forming fibrils on the cell surface. In S. epidermidis proteinaceous biofilm formation can also be mediated by the extracellular matrix binding protein (Embp and S. epidermidis surface protein C (SesC. Additionally, multifunctional proteins such as extracellular adherence protein (Eap and extracellular matrix protein binding protein (Emp of S. aureus and the iron-regulated surface determinant protein C (IsdC of S. lugdunensis can promote biofilm formation in iron-depleted conditions. This multitude of proteins intervene at different stages of biofilm formation with certain proteins contributing to biofilm accumulation and others mediating primary attachment to surfaces. This review examines the contribution of proteins to biofilm formation in staphylococci. The potential to develop vaccines to prevent

  1. Resistance of non-typeable Haemophilus influenzae biofilms is independent of biofilm size.

    Science.gov (United States)

    Reimche, Jennifer L; Kirse, Daniel J; Whigham, Amy S; Swords, W Edward

    2017-02-01

    The inflammatory middle ear disease known as otitis media can become chronic or recurrent in some cases due to failure of the antibiotic treatment to clear the bacterial etiological agent. Biofilms are known culprits of antibiotic-resistant infections; however, the mechanisms of resistance for non-typeable Haemophilus influenzae biofilms have not been completely elucidated. In this study, we utilized in vitro static biofilm assays to characterize clinical strain biofilms and addressed the hypothesis that biofilms with greater biomass and/or thickness would be more resistant to antimicrobial-mediated eradication than thinner and/or lower biomass biofilms. Consistent with previous studies, antibiotic concentrations required to eliminate biofilm bacteria tended to be drastically higher than concentrations required to kill planktonic bacteria. The size characterizations of the biofilms formed by the clinical isolates were compared to their minimum biofilm eradication concentrations for four antibiotics. This revealed no correlation between biofilm thickness or biomass and the ability to resist eradication by antibiotics. Therefore, we concluded that biofilm size does not play a role in antibiotic resistance, suggesting that reduction of antibiotic penetration may not be a significant mechanism for antibiotic resistance for this bacterial opportunist.

  2. Subinhibitory concentrations of azithromycin decrease nontypeable Haemophilus influenzae biofilm formation and Diminish established biofilms.

    Science.gov (United States)

    Starner, Timothy D; Shrout, Joshua D; Parsek, Matthew R; Appelbaum, Peter C; Kim, GunHee

    2008-01-01

    Nontypeable Haemophilus influenzae (NTHi) commonly causes otitis media, chronic bronchitis in emphysema, and early airway infections in cystic fibrosis. Long-term, low-dose azithromycin has been shown to improve clinical outcomes in chronic lung diseases, although the mechanism of action remains unclear. The inhibition of bacterial biofilms by azithromycin has been postulated to be one mechanism mediating these effects. We hypothesized that subinhibitory concentrations of azithromycin would affect NTHi biofilm formation. Laboratory strains of NTHi expressing green fluorescent protein and azithromycin-resistant clinical isolates were grown in flow-cell and static-culture biofilm models. Using a range of concentrations of azithromycin and gentamicin, we measured the degree to which these antibiotics inhibited biofilm formation and persistence. Large biofilms formed over 2 to 4 days in a flow cell, displaying complex structures, including towers and channels. Subinhibitory concentrations of azithromycin significantly decreased biomass and maximal thickness in both forming and established NTHi biofilms. In contrast, subinhibitory concentrations of gentamicin had no effect on biofilm formation. Furthermore, established NTHi biofilms became resistant to gentamicin at concentrations far above the MIC. Biofilm formation of highly resistant clinical NTHi isolates (azithromycin MIC of > 64 microg/ml) was similarly decreased at subinhibitory azithromycin concentrations. Clinically obtainable azithromycin concentrations inhibited biofilms in all but the most highly resistant isolates. These data show that subinhibitory concentrations of azithromycin have antibiofilm properties, provide mechanistic insights, and supply an additional rationale for the use of azithromycin in chronic biofilm infections involving H. influenzae.

  3. Physics of biofilms: the initial stages of biofilm formation and dynamics

    Science.gov (United States)

    Lambert, Guillaume; Bergman, Andrew; Zhang, Qiucen; Bortz, David; Austin, Robert

    2014-04-01

    One of the physiological responses of bacteria to external stress is to assemble into a biofilm. The formation of a biofilm greatly increases a bacterial population's resistance to a hostile environment by shielding cells, for example, from antibiotics. In this paper, we describe the conditions necessary for the emergence of biofilms in natural environments and relate them to the emergence of biofilm formation inside microfluidic devices. We show that competing species of Escherichia coli bacteria form biofilms to spatially segregate themselves in response to starvation stress, and use in situ methods to characterize the physical properties of the biofilms. Finally, we develop a microfluidic platform to study the inter-species interactions and show how biofilm-mediated genetic interactions can improve a species’ resistance to external stress.

  4. Alginate production affects Pseudomonas aeruginosa biofilm development and architecture, but is not essential for biofilm formation

    DEFF Research Database (Denmark)

    Stapper, A.P.; Narasimhan, G.; Oman, D.E.

    2004-01-01

    Extracellular polymers can facilitate the non-specific attachment of bacteria to surfaces and hold together developing biofilms. This study was undertaken to qualitatively and quantitatively compare the architecture of biofilms produced by Pseudomonas aeruginosa strain PAO1 and its alginate......-overproducing (mucA22) and alginate-defective (algD) variants in order to discern the role of alginate in biofilm formation. These strains, PAO1, Alg(+) PAOmucA22 and Alg(-) PAOalgD, tagged with green fluorescent protein, were grown in a continuous flow cell system to characterize the developmental cycles...... of their biofilm formation using confocal laser scanning microscopy. Biofilm Image Processing (BIP) and Community Statistics (COMSTAT) software programs were used to provide quantitative measurements of the two-dimensional biofilm images. All three strains formed distinguishable biofilm architectures, indicating...

  5. Use of an in vitro flat-bed biofilm model to measure biologically active anti-odour compounds.

    Science.gov (United States)

    Saad, S; Hewett, K; Greenman, J

    2013-09-01

    The objective of this study was to demonstrate the utility of a modified flat-bed perfusion biofilm matrix system for testing toothpaste formulations directly, without dilution, as a layer in direct contact with the biofilm matrix surface. Final biofilm yields and volatile sulphur compounds (VSC) biogenesis were measured to show the relative efficacy of toothpaste formulations. Diffusion characteristics of the flat-bed system to exposure with Meridol® tooth and tongue gel (TTG; 1,400 ppm F(-) from amine fluoride/stannous fluoride, 0.5 % zinc lactate, oral malodour counteractives) was assessed using a bioluminescent target species Escherichia coli Nissle 1917/pGLITE coupled with a low-light photon camera to visualise the kill kinetics. Tongue-flora derived, mixed culture biofilms (n = 4) received 5, 15 and 30 min treatment with TTG, respectively, to determine the optimum time of exposure. VSC biogenesis was measured from headspace samples by gas chromatography prior to and following treatment of two daily applications for 4 days of treatment (TTG), positive control (CHX gel) and negative controls (placebo and sham treatment). Viable counts were performed at the end of experiments by destructive sampling of the biofilms and plating onto selective and non-selective agar. Following a single treatment with TTG, the E. coli biofilm with lux target gave >50 % reduction of luminescence within 2 to 3 h before recovering to a steady state over 10 h, suggesting biofilm cidal activity rather biostasis. For mixed culture biofilms, 15- and 30-min treatment exposure with TTG gave almost identical reductions in final biofilm yields. For comparing efficacy of treatments, biofilms treated with TTG gave greatest reductions in both pre-post levels of H2S (P biofilm microflora within a real mouth, including VSC biogenesis and its inhibition by exposure to active agents as components of toothpastes and gels applied in direct contact with the biofilm.

  6. Synergistic effects in mixed Escherichia coli biofilms

    DEFF Research Database (Denmark)

    Reisner, A.; Holler, B.M.; Molin, Søren

    2006-01-01

    the pathways governing development of more complex heterogeneous communities. In this study, we established a laboratory model where biofilm-stimulating effects due to interactions between genetically diverse strains of Escherichia coli were monitored. Synergistic induction of biofilm formation resulting from...

  7. Cellular chain formation in Escherichia coli biofilms

    DEFF Research Database (Denmark)

    Vejborg, Rebecca Munk; Klemm, Per

    2009-01-01

    In this study we report on a novel structural phenotype in Escherichia coli biofilms: cellular chain formation. Biofilm chaining in E. coli K-12 was found to occur primarily by clonal expansion, but was not due to filamentous growth. Rather, chain formation was the result of intercellular...

  8. The 'Swiss cheese' instability of bacterial biofilms

    CERN Document Server

    Jang, Hongchul; Stocker, Roman

    2012-01-01

    We demonstrate a novel pattern that results in bacterial biofilms as a result of the competition between hydrodynamic forces and adhesion forces. After the passage of an air plug, the break up of the residual thin liquid film scrapes and rearranges bacteria on the surface, such that a Swiss cheese pattern of holes is left in the residual biofilm.

  9. Biofilms: The Stronghold of Legionella pneumophila

    Directory of Open Access Journals (Sweden)

    Mena Abdel-Nour

    2013-10-01

    Full Text Available Legionellosis is mostly caused by Legionella pneumophila and is defined as a severe respiratory illness with a case fatality rate ranging from 5% to 80%. L. pneumophila is ubiquitous in natural and anthropogenic water systems. L. pneumophila is transmitted by inhalation of contaminated aerosols produced by a variety of devices. While L. pneumophila replicates within environmental protozoa, colonization and persistence in its natural environment are also mediated by biofilm formation and colonization within multispecies microbial communities. There is now evidence that some legionellosis outbreaks are correlated with the presence of biofilms. Thus, preventing biofilm formation appears as one of the strategies to reduce water system contamination. However, we lack information about the chemical and biophysical conditions, as well as the molecular mechanisms that allow the production of biofilms by L. pneumophila. Here, we discuss the molecular basis of biofilm formation by L. pneumophila and the roles of other microbial species in L. pneumophila biofilm colonization. In addition, we discuss the protective roles of biofilms against current L. pneumophila sanitation strategies along with the initial data available on the regulation of L. pneumophila biofilm formation.

  10. Dental diagnostics: molecular analysis of oral biofilms.

    Science.gov (United States)

    Hiyari, Sarah; Bennett, Katie M

    2011-01-01

    Dental biofilms are complex, multi-species bacterial communities that colonize the mouth in the form of plaque and are known to cause dental caries and periodontal disease. Biofilms are unique from planktonic bacteria in that they are mutualistic communities with a 3-dimensional structure and complex nutritional and communication pathways. The homeostasis within the biofilm colony can be disrupted, causing a shift in the bacterial composition of the colony and resulting in proliferation of pathogenic species. Because of this dynamic lifestyle, traditional microbiological techniques are inadequate for the study of biofilms. Many of the bacteria present in the oral cavity are viable but not culturable, which severely limits laboratory analysis. However, with the advent of new molecular techniques, the microbial makeup of oral biofilms can be better identified. Some of these techniques include DNA-DNA hybridization, 16S rRNA gene sequencing, denaturing gradient gel electrophoresis, terminal restriction fragment length polymorphism, denaturing high-performance liquid chromatography and pyrosequencing. This review provides an overview of biofilm formation and examines the major molecular techniques currently used in oral biofilm analysis. Future applications of the molecular analysis of oral biofilms in the diagnosis and treatment of caries and periodontal disease are also discussed.

  11. 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.

  12. Biofilm ved kronisk rhinosinuitis og cystisk fibrose

    DEFF Research Database (Denmark)

    Fisker, Jacob; Buchwald, Christian von; Johansen, Helle Krogh

    2011-01-01

    Microbial biofilms are known to cause persistent foreign-body infections and have recently been acknowledged as involved in more than 65% of all human infections. Microbial biofilms have been detected in chronic rhinosinusitis, and chronic rhinosinusitis is mandatory in patients with cystic...

  13. Screening of Compounds against Gardnerella vaginalis Biofilms.

    Directory of Open Access Journals (Sweden)

    Cornelia Gottschick

    Full Text Available Bacterial vaginosis (BV is a common infection in reproductive age woman and is characterized by dysbiosis of the healthy vaginal flora which is dominated by Lactobacilli, followed by growth of bacteria like Gardnerella vaginalis. The ability of G. vaginalis to form biofilms contributes to the high rates of recurrence that are typical for BV and which unfortunately make repeated antibiotic therapy inevitable. Here we developed a biofilm model for G. vaginalis and screened a large spectrum of compounds for their ability to prevent biofilm formation and to resolve an existing G. vaginalis biofilm. The antibiotics metronidazole and tobramycin were highly effective in preventing biofilm formation, but had no effect on an established biofilm. The application of the amphoteric tenside sodium cocoamphoacetate (SCAA led to disintegration of existing biofilms, reducing biomass by 51% and viability by 61% and it was able to increase the effect of metronidazole by 40% (biomass and 61% (viability. Our data show that attacking the biofilm and the bacterial cells by the combination of an amphoteric tenside with the antibiotic metronidazole might be a useful strategy against BV.

  14. Ciliates as engineers of phototrophic biofilms.

    NARCIS (Netherlands)

    Weerman, E.J.; Geest, H.G.; Meulen, M.D.; Manders, E.M.M.; Van de Koppel, J.; Herman, P.M.J.; Admiraal, W.

    2011-01-01

    1.Phototrophic biofilms consist of a matrix of phototrophs, non-photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the ‘engineering’ effects

  15. Ciliates as engineers of phototrophic biofilms.

    NARCIS (Netherlands)

    Weerman, E.J.; van der Geest, H.G.; van der Meulen, M.D; Manders, E.M.M.; van de Koppel, J.; Herman, P.M.J.; Admiraal, W.

    2011-01-01

    1. Phototrophic biofilms consist of a matrix of phototrophs, non-photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the ‘engineering’ effect

  16. Introduction to Biofilms Thematic Minireview Series.

    Science.gov (United States)

    Allewell, Norma M

    2016-06-10

    The biofilms that many bacteria and fungi produce enable them to form communities, adhere tightly to surfaces, evade host immunity, and resist antibiotics. Pathogenic microorganisms that form biofilms are very difficult to eradicate and thus are a frequent source of life-threatening, hospital-acquired infections. This series of five minireviews from the Journal of Biological Chemistry provides a broad overview of our current understanding of biofilms and the challenges that remain. The structure, biosynthesis, and biological function of the biofilms produced by pathogenic fungi are the subject of the first article, by Sheppard and Howell. Gunn, Bakaletz, and Wozniak focus on the biochemistry and structure of bacterial biofilms, how these structures enable bacteria to evade host immunity, and current and developing strategies for overcoming this resistance. The third and fourth articles present two of the best understood cell signaling pathways involved in biofilm formation. Valentini and Filloux focus on cyclic di-GMP, while Kavanaugh and Horswill discuss the quorum-sensing (agr) system and the relationship between quorum sensing and biofilm formation. Mechanisms of antibiotic resistance, particularly the role of efflux pumps and the development of persister cells, are the topics of the final article by Van Acker and Coenye. The advances described in this series guarantee that ongoing interdisciplinary and international efforts will lead to new insights into the basic biology of biofilm formation, as well as new strategies for therapeutic interventions.

  17. Transferrin Impacts Bacillus thuringiensis Biofilm Levels

    Directory of Open Access Journals (Sweden)

    Bianca Garner

    2016-01-01

    Full Text Available The present study examined the impact of transferrin on Bacillus thuringiensis biofilms. Three commercial strains, an environmental strain (33679, the type strain (10792, and an isolate from a diseased insect (700872, were cultured in iron restricted minimal medium. All strains produced biofilm when grown in vinyl plates at 30°C. B. thuringiensis 33679 had a biofilm biomass more than twice the concentration exhibited by the other strains. The addition of transferrin resulted in slightly increased growth yields for 2 of the 3 strains tested, including 33679. In contrast, the addition of 50 μg/mL of transferrin resulted in an 80% decrease in biofilm levels for strain 33679. When the growth temperature was increased to 37°C, the addition of 50 μg/mL of transferrin increased culture turbidity for only strain 33679. Biofilm levels were again decreased in strain 33679 at 37°C. Growth of B. thuringiensis cultures in polystyrene resulted in a decrease in overall growth yields at 30°C, with biofilm levels significantly decreased for 33679 in the presence of transferrin. These findings demonstrate that transferrin impacts biofilm formation in select strains of B. thuringiensis. Identification of these differences in biofilm regulation may be beneficial in elucidating potential virulence mechanisms among the differing strains.

  18. Investigations on the use of anodic stripping voltammetry for the analyses of lead in saline environments

    Energy Technology Data Exchange (ETDEWEB)

    Case, C.W.

    1978-08-01

    Research is reported directed to modification of the anodic stripping voltammetry (ASV) analytic method in order to acquire data for lead from ambient sea water conditions, and development of a chemical model which uses these data to identify inorganic lead species for saline environments. Laboratory and field samples were analyzed for lead partitioning in: (a) KCl electrolyte solutions; (b) I.A.P.S.O. Standard Sea Water; (c) seawater samples from Quatsino Sound, British Columbia; (d) a series of seawater samples from San Francisco Bay; and (e) seawater samples from the Gulf of Mexico. The electrochemical traits of the lead species and the ASV oxidation potential expression are the fundamental constituents of the chemical model. The model uses the data from the analyses to provide the mass balance relationships for lead partitioned among the major anions in seawater. The laboratory analyses of KCl electrolyte and Standard Seawater give the following results. The modified ASV method and chemical model provide information on ambient labile and non-labile inorganic lead complexes in these saline solutions down to the parts-per-billion level. No purge and the simple electrodes cause some erratic behavior and spurious potentials, but the data are reproducible. In addition to Pb/sup 2 +/, the most dominant measured lead species in order include PbCO/sub 3//sup 0/, PbSO/sub 4//sup 0/, PbCl/sup +/, and Pb(NO/sub 3/)/sub 2//sup 0/ from the lead additions solution. The analyses of the field samples give the following results. Samples were taken from the partially anoxic basin in Quatsino Sound, British Columbia with one successful analysis which is for somewhat normal dissolved oxygen conditions. Data show that lead is partitioned among Pb/sup 2 +/, Pb(OH)/sub 2//sup 0/, PbCO/sub 3//sup 0/, and PbSO/sub 4//sup 0/. The analyses with purge for the San Francisco Bay water partitions lead among Pb/sup 2 +/, PbCO/sub 3//sup 0/, PbSO/sub 4//sup 0/, PbCl/sup +/, and Pb

  19. Oral biofilms: emerging concepts in microbial ecology.

    Science.gov (United States)

    Filoche, S; Wong, L; Sissons, C H

    2010-01-01

    Oral biofilms develop under a range of different conditions and different environments. This review will discuss emerging concepts in microbial ecology and how they relate to oral biofilm development and the treatment of oral diseases. Clues to how oral biofilms develop may lie in other complex systems, such as interactions between host and gut microbiota, and even in factors that affect biofilm development on leaf surfaces. Most of the conditions under which oral biofilms develop are tightly linked to the overall health and biology of the host. Advances in molecular techniques have led to a greater appreciation of the diversity of human microbiota, the extent of interactions with the human host, and how that relates to inter-individual variation. As a consequence, plaque development may no longer be thought of as a generic process, but rather as a highly individualized process, which has ramifications for the treatment of the diseases it causes.

  20. Role of multicellular aggregates in biofilm formation

    DEFF Research Database (Denmark)

    Kragh, Kasper N.; Hutchison, Jaime B.; Melaugh, Gavin

    2016-01-01

    In traditional models of in vitro biofilm development, individual bacterial cells seed a surface, multiply, and mature into multicellular, three-dimensional structures. Much research has been devoted to elucidating the mechanisms governing the initial attachment of single cells to surfaces. However......, in natural environments and during infection, bacterial cells tend to clump as multicellular aggregates, and biofilms can also slough off aggregates as a part of the dispersal process. This makes it likely that biofilms are often seeded by aggregates and single cells, yet how these aggregates impact biofilm...... initiation and development is not known. Here we use a combination of experimental and computational approaches to determine the relative fitness of single cells and preformed aggregates during early development of Pseudomonas aeruginosa biofilms. We find that the relative fitness of aggregates depends...

  1. The Physics of Biofilms -- An Introduction

    CERN Document Server

    Mazza, Marco G

    2016-01-01

    Biofilms are complex, self-organized consortia of microorganisms that produce a functional, protective matrix of biomolecules. Physically, the structure of a biofilm can be described as an entangled polymer network which grows and changes under the effect of gradients of nutrients, cell differentiation, quorum sensing, bacterial motion, and interaction with the environment. Its development is complex, and constantly adapting to environmental stimuli. Here, we review the fundamental physical processes the govern the inception, growth and development of a biofilm. Two important mechanisms guide the initial phase in a biofilm life cycle: (\\emph{i}) the cell motility near or at a solid interface, and (\\emph{ii}) the cellular adhesion. Both processes are crucial for initiating the colony and for ensuring its stability. A mature biofilm behaves as a viscoelastic fluid with a complex, history-dependent dynamics. We discuss progress and challenges in the determination of its physical properties. Experimental and theo...

  2. The ecology and biogeochemistry of stream biofilms.

    Science.gov (United States)

    Battin, Tom J; Besemer, Katharina; Bengtsson, Mia M; Romani, Anna M; Packmann, Aaron I

    2016-04-01

    Streams and rivers form dense networks, shape the Earth's surface and, in their sediments, provide an immensely large surface area for microbial growth. Biofilms dominate microbial life in streams and rivers, drive crucial ecosystem processes and contribute substantially to global biogeochemical fluxes. In turn, water flow and related deliveries of nutrients and organic matter to biofilms constitute major constraints on microbial life. In this Review, we describe the ecology and biogeochemistry of stream biofilms and highlight the influence of physical and ecological processes on their structure and function. Recent advances in the study of biofilm ecology may pave the way towards a mechanistic understanding of the effects of climate and environmental change on stream biofilms and the biogeochemistry of stream ecosystems.

  3. 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...

  4. 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...

  5. A Subinhibitory Concentration of Clarithromycin Inhibits Mycobacterium avium Biofilm Formation

    OpenAIRE

    2004-01-01

    Mycobacterium avium causes disseminated infection in immunosuppressed individuals and lung infection in patients with chronic lung diseases. M. avium forms biofilm in the environment and possibly in human airways. Antibiotics with activity against the bacterium could inhibit biofilm formation. Clarithromycin inhibits biofilm formation but has no activity against established biofilm.

  6. Discovering Biofilms: Inquiry-Based Activities for the Classroom

    Science.gov (United States)

    Redelman, Carly V.; Marrs, Kathleen; Anderson, Gregory G.

    2012-01-01

    In nature, bacteria exist in and adapt to different environments by forming microbial communities called "biofilms." We propose simple, inquiry-based laboratory exercises utilizing a biofilm formation assay, which allows controlled biofilm growth. Students will be able to qualitatively assess biofilm growth via staining. Recently, we developed a…

  7. In vitro phenotypic differentiation towards commensal and pathogenic oral biofilms

    NARCIS (Netherlands)

    Janus, M.M.; Keijser, B.J.F.; Bikker, F.J.; Exterkate, R.A.M.; Crielaard, W.; Krom, B.P.

    2015-01-01

    Commensal oral biofilms, defined by the absence of pathology-related phenotypes, are ubiquitously present. In contrast to pathological biofilms commensal biofilms are rarely studied. Here, the effect of the initial inoculum and subsequent growth conditions on in vitro oral biofilms was studied. Biof

  8. Biofilms On Orbit and On Earth: Current Methods, Future Needs

    Science.gov (United States)

    Vega, Leticia

    2013-01-01

    Biofilms have played a significant role on the effectiveness of life support hardware on the Space Shuttle and International Space Station (ISS). This presentation will discuss how biofilms impact flight hardware, how on orbit biofilms are analyzed from an engineering and research perspective, and future needs to analyze and utilize biofilms for long duration, deep space missions.

  9. Use of a modified, high-sensitivity, anodic stripping voltammetry method for determination of zinc speciation in the North Atlantic Ocean.

    Science.gov (United States)

    Jakuba, Rachel Wisniewski; Moffett, James W; Saito, Mak A

    2008-05-05

    Zinc speciation is considered to be an important determinant of the biological availability of zinc. Yet in oceanic surface waters, characterization of zinc speciation is difficult due to the low concentrations of this essential micronutrient. In this study, an anodic stripping voltammetry method previously developed for the total determination of cadmium and lead was successfully adapted to the measurement of zinc speciation. The method differs from previous zinc speciation anodic stripping voltammetry methods in that a fresh mercury film is plated with each sample aliquot. The fresh film anodic stripping voltammetry method was compared to competitive ligand exchange cathodic stripping voltammetry in a profile from the North Atlantic Ocean. Results using the fresh film anodic stripping voltammetry method were similar to those determined using the cathodic stripping voltammetry method, though ligand concentrations determined by fresh film anodic stripping voltammetry were generally slightly higher than those determined by cathodic stripping voltammetry. There did not seem to be a systematic difference between methods for the estimates of conditional stability constants. The ligand concentration in the North Atlantic profile ranged from 0.9 to 1.5 nmol L(-1) as determined by fresh film anodic stripping voltammetry and 0.6 to 1.3 nmol L(-1) as determined by cathodic stripping voltammetry. The conditional stability constants determined by fresh film anodic stripping voltammetry were 10(9.8)-10(10.5) and by cathodic stripping voltammetry were 10(9.8)-10(11.3).

  10. Waste water derived electroactive microbial biofilms: growth, maintenance, and basic characterization.

    Science.gov (United States)

    Gimkiewicz, Carla; Harnisch, Falk

    2013-12-29

    The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup controlled by a potentiostat. Thereby the use of potentiostats allows an exact adjustment of the electrode potential and ensures reproducible microbial culturing conditions. During growth the current production is monitored using chronoamperometry (CA). Based on these data the maximum current density (jmax) and the coulombic efficiency (CE) are discussed as measures for characterization of the bioelectrocatalytic performance. Cyclic voltammetry (CV), a nondestructive, i.e. noninvasive, method, is used to study the extracellular electron transfer (EET) of electroactive bacteria. CV measurements are performed on anodic biofilm electrodes in the presence of the microbial substrate, i.e. turnover conditions, and in the absence of the substrate, i.e. nonturnover conditions, using different scan rates. Subsequently, data analysis is exemplified and fundamental thermodynamic parameters of the microbial EET are derived and explained: peak potential (Ep), peak current density (jp), formal potential (E(f)) and peak separation (ΔEp). Additionally the limits of the method and the state-of the art data analysis are addressed. Thereby this video-article shall provide a guide for the basic experimental steps and the fundamental data analysis.

  11. Formation of biofilms in drinking water distribution networks, a case study in two cities in Finland and Latvia.

    Science.gov (United States)

    Lehtola, Markku J; Juhna, Tālis; Miettinen, Ilkka T; Vartiainen, Terttu; Martikainen, Pertti J

    2004-12-01

    The formation of biofilms in drinking water distribution networks is a significant technical, aesthetic and hygienic problem. In this study, the effects of assimilable organic carbon, microbially available phosphorus (MAP), residual chlorine, temperature and corrosion products on the formation of biofilms were studied in two full-scale water supply systems in Finland and Latvia. Biofilm collectors consisting of polyvinyl chloride pipes were installed in several waterworks and distribution networks, which were supplied with chemically precipitated surface waters and groundwater from different sources. During a 1-year study, the biofilm density was measured by heterotrophic plate counts on R2A-agar, acridine orange direct counting and ATP-analyses. A moderate level of residual chlorine decreased biofilm density, whereas an increase of MAP in water and accumulated cast iron corrosion products significantly increased biofilm density. This work confirms, in a full-scale distribution system in Finland and Latvia, our earlier in vitro finding that biofilm formation is affected by the availability of phosphorus in drinking water.

  12. The inhibitory effect of Thymus vulgaris extracts on the planktonic form and biofilm structures of six human pathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Zinab Mohsenipour

    2015-06-01

    Full Text Available Objective: Microorganisms are responsible for many problems in industry and medicine because of biofilm formation. Therefore, this study was aimed to examine the effect of Thymus vulgaris (T. vulgaris extracts on the planktonic form and biofilm structures of six pathogenic bacteria. Materials and methods: Antimicrobial activities of the plant extracts against the planktonic form of the bacteria were determined using the disc diffusion method. MIC and MBC values were evaluated using macrobroth dilution technique. Anti-biofilm effects were assessed by microtiter plate method. Results: According to disc diffusion test (MIC and MBC, the ability of Thymus vulgaris (T. vulgaris extracts for inhibition of bacteria in planktonic form was confirmed. In dealing with biofilm structures, the inhibitory effect of the extracts was directly correlated to their concentration. Except for the inhibition of biofilm formation, efficacy of each extract was independent from type of solvent. Conclusion: According to the potential of Thymus vulgaris (T. vulgaris extracts to inhibit the test bacteria in planktonic and biofilm form, it can be suggested that Thymus vulgaris(T. vulgaris extracts can be applied as antimicrobial agents against the pathogenic bacteria particularly in biofilm forms.

  13. Maltodextrin enhances biofilm elimination by electrochemical scaffold.

    Science.gov (United States)

    Sultana, Sujala T; Call, Douglas R; Beyenal, Haluk

    2016-10-26

    Electrochemical scaffolds (e-scaffolds) continuously generate low concentrations of H2O2 suitable for damaging wound biofilms without damaging host tissue. Nevertheless, retarded diffusion combined with H2O2 degradation can limit the efficacy of this potentially important clinical tool. H2O2 diffusion into biofilms and bacterial cells can be increased by damaging the biofilm structure or by activating membrane transportation channels by exposure to hyperosmotic agents. We hypothesized that e-scaffolds would be more effective against Acinetobacter baumannii and Staphylococcus aureus biofilms in the presence of a hyperosmotic agent. E-scaffolds polarized at -600 mVAg/AgCl were overlaid onto preformed biofilms in media containing various maltodextrin concentrations. E-scaffold alone decreased A. baumannii and S. aureus biofilm cell densities by (3.92 ± 0.15) log and (2.31 ± 0.12) log, respectively. Compared to untreated biofilms, the efficacy of the e-scaffold increased to a maximum (8.27 ± 0.05) log reduction in A. baumannii and (4.71 ± 0.12) log reduction in S. aureus biofilm cell densities upon 10 mM and 30 mM maltodextrin addition, respectively. Overall ~55% decrease in relative biofilm surface coverage was achieved for both species. We conclude that combined treatment with electrochemically generated H2O2 from an e-scaffold and maltodextrin is more effective in decreasing viable biofilm cell density.

  14. Maltodextrin enhances biofilm elimination by electrochemical scaffold

    Science.gov (United States)

    Sultana, Sujala T.; Call, Douglas R.; Beyenal, Haluk

    2016-01-01

    Electrochemical scaffolds (e-scaffolds) continuously generate low concentrations of H2O2 suitable for damaging wound biofilms without damaging host tissue. Nevertheless, retarded diffusion combined with H2O2 degradation can limit the efficacy of this potentially important clinical tool. H2O2 diffusion into biofilms and bacterial cells can be increased by damaging the biofilm structure or by activating membrane transportation channels by exposure to hyperosmotic agents. We hypothesized that e-scaffolds would be more effective against Acinetobacter baumannii and Staphylococcus aureus biofilms in the presence of a hyperosmotic agent. E-scaffolds polarized at −600 mVAg/AgCl were overlaid onto preformed biofilms in media containing various maltodextrin concentrations. E-scaffold alone decreased A. baumannii and S. aureus biofilm cell densities by (3.92 ± 0.15) log and (2.31 ± 0.12) log, respectively. Compared to untreated biofilms, the efficacy of the e-scaffold increased to a maximum (8.27 ± 0.05) log reduction in A. baumannii and (4.71 ± 0.12) log reduction in S. aureus biofilm cell densities upon 10 mM and 30 mM maltodextrin addition, respectively. Overall ~55% decrease in relative biofilm surface coverage was achieved for both species. We conclude that combined treatment with electrochemically generated H2O2 from an e-scaffold and maltodextrin is more effective in decreasing viable biofilm cell density. PMID:27782161

  15. Head-to-head comparisons of carbon fiber microelectrode coatings for sensitive and selective neurotransmitter detection by voltammetry.

    Science.gov (United States)

    Singh, Yogesh S; Sawarynski, Lauren E; Dabiri, Pasha D; Choi, Wonwoo R; Andrews, Anne M

    2011-09-01

    Voltammetry is widely used to investigate neurotransmission and other biological processes but is limited by poor chemical selectivity and fouling of commonly used carbon fiber microelectrodes (CFMs). We performed direct comparisons of three key coating materials purported to impart selectivity and fouling resistance to electrodes: Nafion, base-hydrolyzed cellulose acetate (BCA), and fibronectin. We systematically evaluated the impact on a range of electrode parameters. Fouling due to exposure to brain tissue was investigated using an approach that minimizes the use of animals while enabling evaluation of statistically significant populations of electrodes. We find that BCA is relatively fouling-resistant. Moreover, detection at BCA-coated CFMs can be tuned by altering hydrolysis times to minimize the impact on sensitivity losses while maintaining fouling resistance. Fibronectin coating is associated with moderate losses in sensitivity after coating and fouling. Nafion imparts increased sensitivity for dopamine and norepinephrine but not serotonin, as well as the anticipated selectivity for cationic neurotransmitters over anionic metabolites. Although Nafion has been suggested to resist fouling, both dip-coating and electrodeposition of Nafion are associated with substantial fouling, similar to levels observed at bare electrodes after exposure to brain tissue. Direct comparisons of these coatings identified unique electroanalytical properties of each that can be used to guide selection tailored to the goals and environment of specific studies.

  16. Combined Reactor and Microelectrode Measurements in Laboratory Grown Biofilms

    DEFF Research Database (Denmark)

    Larsen, Tove; Harremoës, Poul

    1994-01-01

    A combined biofilm reactor-/microelectrode experimental set-up has been constructed, allowing for simultaneous reactor mass balances and measurements of concentration profiles within the biofilm. The system consists of an annular biofilm reactor equipped with an oxygen microelectrode. Experiments...... were carried out with aerobic glucose and starch degrading biofilms. The well described aerobic glucose degradation biofilm system was used to test the combined reactor set-up. Results predicted from known biofilm kinetics were obtained. In the starch degrading biofilm, basic assumptions were tested...

  17. Electrochemical-Voltammetry Behavior of Several Aromatic Aldehydes in Acid Solution

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The electrochemical-voltammetry behavior of vanillin, heliotropin, anisaldehyde on the surface ofPt, Au electrodes in acid solution has been studied by means of the electrochemical cyclic voltammetry method. I was found that the electrochemical processes of them are irreversible on both Pt and Au elec-trodes. The electrochemical activity of vanillin is stronger than heliotropin's and heliotropin's is stronger than, anisaldehyde's on Pt electrode. While the electrochemical activity of anisaldehyde is stronger than heliotropin's and vanillin's is the weakest on Au. The results indicate that when they are used as additives for electroplating, they must be consumptive, and it will improve the leveling ability of plating solution and brightness of the deposition layer.

  18. Determination of the wine preservative sulphur dioxide with cyclic voltammetry using inkjet printed electrodes.

    Science.gov (United States)

    Schneider, Marion; Türke, Alexander; Fischer, Wolf-Joachim; Kilmartin, Paul A

    2014-09-15

    During winemaking sulphur dioxide is added to prevent undesirable reactions. However, concerns over the harmful effects of sulphites have led to legal limits being placed upon such additives. There is thus a need for simple and selective determinations of sulphur dioxide in wine, especially during winemaking. The simultaneous detection of polyphenols and sulphur dioxide, using cyclic voltammetry at inert electrodes is challenging due to close oxidation potentials. In the present study, inkjet printed electrodes were developed with a suitable voltammetric signal on which the polyphenol oxidation is suppressed and the oxidation peak height for sulphur dioxide corresponds linearly to the concentration. Different types of working electrodes were printed. Electrodes consisting of gold nanoparticles mixed with silver showed the highest sensitivity towards sulphur dioxide. Low cost production of the sensor elements and ultra fast determination of sulphur dioxide by cyclic voltammetry makes this technique very promising for the wine industry.

  19. Anodic stripping voltammetry of silver nanoparticles: aggregation leads to incomplete stripping.

    Science.gov (United States)

    Cloake, Samantha J; Toh, Her Shuang; Lee, Patricia T; Salter, Chris; Johnston, Colin; Compton, Richard G

    2015-02-01

    The influence of nanoparticle aggregation on anodic stripping voltammetry is reported. Dopamine-capped silver nanoparticles were chosen as a model system, and melamine was used to induce aggregation in the nanoparticles. Through the anodic stripping of the silver nanoparticles that were aggregated to different extents, it was found that the peak area of the oxidative signal corresponding to the stripping of silver to silver(I) ions decreases with increasing aggregation. Aggregation causes incomplete stripping of the silver nanoparticles. Two possible mechanisms of 'partial oxidation' and 'inactivation' of the nanoparticles are proposed to account for this finding. Aggregation effects must be considered when anodic stripping voltammetry is used for nanoparticle detection and quantification. Hence, drop casting, which is known to lead to aggregation, is not encouraged for preparing electrodes for analytical purposes.

  20. Facilitated ion transfer of protonated primary organic amines studied by square wave voltammetry and chronoamperometry.

    Science.gov (United States)

    Torralba, E; Ortuño, J A; Molina, A; Serna, C; Karimian, F

    2014-05-15

    The transfer of the protonated forms of heptylamine, octylamine, decylamine, procaine and procainamide facilitated by dibenzo-18-crown-6 from water to a solvent polymeric membrane has been investigated by using cyclic square wave voltammetry. The experimental voltammograms obtained are in good agreement with theoretical predictions. The values of the standard ion transfer potential, complexation constant and diffusion coefficient in water have been obtained from these experiments, and have been used to draw some conclusions about the lipophilicity of these species and the relative stability of the organic ammonium complexes with dibenzo-18-crown-6. The results have been compared with those provided by linear sweep voltammetry. Calibration graphs were obtained with both techniques. An interesting chronoamperometric method for the determination of the diffusion coefficient of the target ion in the membrane has been developed and applied to all these protonated amines.

  1. EC-FORC: A New Cyclic Voltammetry Based Method for Examining Phase Transitions and Predicting Equilibrium

    CERN Document Server

    Hamad, Ibrahim Abou; Novotny, Mark A; Rikvold, Per Arne

    2007-01-01

    We propose a new, cyclic-voltammetry based experimental technique that can not only differentiate between discontinuous and continuous phase transitions in an adsorbate layer, but also quite accurately recover equilibrium behavior from dynamic analysis of systems with a continuous phase transition. The Electrochemical first-order reversal curve (EC-FORC) diagram for a discontinuous phase transition (nucleation and growth), such as occurs in underpotential deposition, is characterized by a negative region, while such a region does not exist for a continuous phase transition, such as occurs in the electrosorption of Br on Ag(100). Moreover, for systems with a continuous phase transition, the minima of the individual EC-FORCs trace the equilibrium curve, even at very high scan rates. Since obtaining experimental data for the EC-FORC method would require only a simple reprogramming of the potentiostat used in conventional cyclic-voltammetry experiments, we believe that this method has significant potential for ea...

  2. New Cyclic Voltammetry Method for Examining Phase Transitions on Electrodes: Simulated Results

    Science.gov (United States)

    Abou Hamad, Ibrahim; Robb, Daniel; Rikvold, Per Arne

    2007-03-01

    We propose a new experimental technique for cyclic voltammetry, based on the first-order reversal curve (FORC) method for analysis of systems undergoing hysteresis. The advantages of this technique are demonstrated by applying it to dynamical models of electrochemical adsorption. The method can not only differentiate between discontinuous and continuous phase transitions, but can also quite accurately recover equilibrium behavior from dynamic analysis of systems with a continuous phase transition. The FORC diagram for a discontinuous phase transition is characterized by a negative (unstable) region separating two positive (stable) regions, while such a negative region does not exist for continuous phase transitions. Experimental data for Electrochemical FORC (EC-FORC) analysis could easily be obtained by simple reprogramming of a potentiostat designed for conventional cyclic-voltammetry experiments.I. Abou Hamad, D.T. Robb, P.A. Rikvold, J. Electroanal. Chem., in press.

  3. Detection of food additives by voltammetry at the liquid-liquid interface.

    Science.gov (United States)

    Herzog, Grégoire; Kam, Victor; Berduque, Alfonso; Arrigan, Damien W M

    2008-06-25

    Electrochemistry at the liquid-liquid interface enables the detection of nonredoxactive species with electroanalytical techniques. In this work, the electrochemical behavior of two food additives, aspartame and acesulfame K, was investigated. Both ions were found to undergo ion-transfer voltammetry at the liquid-liquid interface. Differential pulse voltammetry was used for the preparation of calibration curves over the concentration range of 30-350 microM with a detection limit of 30 microM. The standard addition method was applied to the determination of their concentrations in food and beverage samples such as sweeteners and sugar-free beverages. Selective electrochemically modulated liquid-liquid extraction of these species in both laboratory solutions and in beverage samples was also demonstrated. These results indicate the suitability of liquid-liquid electrochemistry as an analytical approach in food analysis.

  4. Voltametria de onda quadrada. Segunda parte: aplicações Square wave voltammetry. Second part: applications

    Directory of Open Access Journals (Sweden)

    Djenaine de Souza

    2004-10-01

    Full Text Available The aim of this work is to discuss some selected applications of square wave voltammetry published in the last five years. The applications focused here cover several electroanalytical fields such as: determination of pesticides; molecules with biological activity; metals and other environmental pollutants. Special attention is given to the work developed in the Grupo de Materiais Eletroquímicos e Métodos Eletroanalíticos - IQSC - USP concerning the utilization of square wave voltammetry, with different kinds of electrodes, for the determination of pesticides in natural waters and active principles in pharmaceutical formulations. The new methodology is simple, fast and sensitive when compared with the traditional ones such as chromatography and spectrophotometry. The satisfactory results obtained provide alternative procedures for the quality control of drugs and the monitoring of pesticides in natural environments.

  5. CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein, CsrA.

    Directory of Open Access Journals (Sweden)

    Stephan P Willias

    Full Text Available The natural transmission of Yersinia pestis is reliant upon biofilm blockage of the flea vector. However, the environmentally-responsive adaptive regulators which facilitate Y. pestis biofilm production in accordance with the flea midgut milieu are not well understood. We seek to establish the impact of available carbon source metabolism and storage upon Y. pestis biofilm production. Our findings demonstrate that Y. pestis biofilm production is subject to carbon catabolite regulation in which the presence of glucose impairs biofilm production; whereas, the sole metabolism of alternate carbon sources promotes robust biofilm formation. This observation is facilitated by the cAMP receptor protein, CRP. In accordance with a stark growth defect, deletion of crp in both CO92 and KIM6+ Y. pestis strains significantly impaired biofilm production when solely utilizing alternate carbon sources. Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production. Furthermore, CRP did not alter mRNA abundance of previously-characterized hms biofilm synthesis and regulation factors. Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile. Additionally, we assessed the impact of the carbon storage regulator protein, CsrA, upon Y. pestis biofilm production. Contrary to what has been described for E. coli, Y. pestis biofilm formation was found to be enhanced by CsrA. Regardless of media composition and available carbon source, deletion of csrA significantly impaired Y. pestis biofilm production. CsrA was found to promote Y. pestis biofilm production independent of glycogen regulation. Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance. However, deletion of hmsP in the csrA-deficient mutant enabled excessive biofilm production

  6. 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.

  7. Plaque biofilms: the effect of chemical environment on natural human plaque biofilm architecture.

    Science.gov (United States)

    Robinson, C; Strafford, S; Rees, G; Brookes, S J; Kirkham, J; Shore, R C; Watson, P S; Wood, S

    2006-11-01

    The architecture of microbial biofilms especially the outer regions have an important influence on the interaction between biofilm and local environment particularly on the flux of materials into and out of biofilm compartments and as a consequence, biofilm metabolic behaviour. In the case of dental plaque biofilms, architecture will determine access of nutrients including acidogenic substrates and therapeutic materials to the microbial biomass and to the underlying tooth surface. Manipulation of this architecture may offer a means of altering mass transfer into the whole biofilm and biomass and raises the possibility of improving access of therapeutics. Plaque biofilms formed in vivo on human enamel were subjected to a number of different chemical conditions while under observation by confocal laser scanning microscopy in reflection mode. In this way the outer 50-100 microm or so of the biofilms was examined. Density and distribution of biomass were recorded as degree of reflectance. The amount and density of biofilm biomass increased from the plaque saliva interface towards the interior. Plaque biofilms were robust and little affected by mechanical manipulation, high ionic strength or low pH (2.5). Detergent (SLS), however, often appeared to either remove biomass and/or dramatically reduce its density.

  8. A mucosal model to study microbial biofilm development and anti-biofilm therapeutics

    Science.gov (United States)

    Anderson, Michele J.; Parks, Patrick J.; Peterson, Marnie L.

    2013-01-01

    Biofilms are a sessile colony of bacteria which adhere to and persist on surfaces. The ability of bacteria to form biofilms is considered a virulence factor, and in fact is central to the pathogenesis of some organisms. Biofilms are inherently resistant to chemotherapy and host immune responses. Clinically, biofilms are considered a primary cause of a majority of infections, such as otitis media, pneumonia in cystic fibrosis patients and endocarditis. However, the vast majority of the data on biofilm formation comes from traditional microtiter-based or flow displacement assays with no consideration given to host factors. These assays, which have been a valuable tool in high-throughput screening for biofilm-related factors, do not mimic a host-pathogen interaction and may contribute to an inappropriate estimation of the role of some factors in clinical biofilm formation. We describe the development of a novel ex vivo model of biofilm formation on a mucosal surface by an important mucosal pathogen, methicillin resistant S. aureus (MRSA). This model is being used for the identification of microbial virulence factors important in mucosal biofilm formation and novel anti-biofilm therapies. PMID:23246911

  9. Inactivation of Efflux Pumps Abolishes Bacterial Biofilm Formation

    DEFF Research Database (Denmark)

    Kvist, Malin; Hancock, Viktoria; Klemm, Per

    2008-01-01

    Bacterial biofilms cause numerous problems in health care and industry; notably, biofilms are associated with a large number of infections. Biofilm-dwelling bacteria are particularly resistant to antibiotics, making it hard to eradicate biofilm-associated infections. Bacteria rely on efflux pumps...... to get rid of toxic substances. We discovered that efflux pumps are highly active in bacterial biofilms, thus making efflux pumps attractive targets for antibiofilm measures. A number of efflux pump inhibitors (EPIs) are known. EPIs were shown to reduce biofilm formation, and in combination they could...... abolish biofilm formation completely. Also, EPIs were able to block the antibiotic tolerance of biofilms. The results of this feasibility study might pave the way for new treatments for biofilm-related infections and may be exploited for prevention of biofilms in general....

  10. Intrigues of biofilm: A perspective in veterinary medicine.

    Science.gov (United States)

    Abdullahi, Umar Faruk; Igwenagu, Ephraim; Mu'azu, Anas; Aliyu, Sani; Umar, Maryam Ibrahim

    2016-01-01

    Biofilm has a tremendous impact in the field of veterinary medicine, especially the livestock industry, leading to a serious economic loss. Over the years, little attention has been given to biofilm in animals with most of the research geared toward human biofilm diseases. The greatest challenge posed by biofilm is in its incredible ability to resist most of the currently existing antibiotics. This mystery can best be demystified through understanding the mechanism of the quorum sensing which regulate the pathophysiology of biofilm. Ability of biofilm formation in a variety of inanimate surfaces such as animal food contact surfaces is responsible for a host of biofilm diseases affecting animals and humans. In this review, we highlighted some of the challenges of biofilm in livestock and food industries. Also highlighted are; mechanisms of biofilm development, best diagnostic approach and possible novel therapeutic measures needed to combat the menace of biofilm in veterinary medicine.

  11. Cathodic stripping voltammetry of trace Mn(II) at carbon film electrodes

    OpenAIRE

    Filipe, Olga M. S.; Brett,Christopher M.A.

    2003-01-01

    A sensitive voltammetric method is presented for the determination of tract levels of Mn (II) using carbon film electrodes fabricated from carbon resistors of 2 [Omega]. Determination of manganese was made by square wave cathodic stripping voltammetry (CSV), with deposition of manganese as manganese dioxide. Chronoamperometric experiments were made to study MnO2 nucleation and growth. As a result, it was found to be necessary to perform electrode conditioning at a more positive potential to i...

  12. A pipette-based calibration system for fast-scan cyclic voltammetry with fast response times.

    Science.gov (United States)

    Ramsson, Eric S

    2016-01-01

    Fast-scan cyclic voltammetry (FSCV) is an electrochemical technique that utilizes the oxidation and/or reduction of an analyte of interest to infer rapid changes in concentrations. In order to calibrate the resulting oxidative or reductive current, known concentrations of an analyte must be introduced under controlled settings. Here, I describe a simple and cost-effective method, using a Petri dish and pipettes, for the calibration of carbon fiber microelectrodes (CFMs) using FSCV.

  13. EC-FORC: A New Cyclic Voltammetry Based Method for Examining Phase Transitions and Predicting Equilibrium

    OpenAIRE

    2007-01-01

    We propose a new, cyclic-voltammetry based experimental technique that can not only differentiate between discontinuous and continuous phase transitions in an adsorbate layer, but also quite accurately recover equilibrium behavior from dynamic analysis of systems with a continuous phase transition. The Electrochemical first-order reversal curve (EC-FORC) diagram for a discontinuous phase transition (nucleation and growth), such as occurs in underpotential deposition, is characterized by a neg...

  14. Hairy carbon electrodes studied by cyclic voltammetry and battery discharge testing

    Science.gov (United States)

    Chung, Deborah D. L.; Shui, Xiaoping; Frysz, Christine A.

    1993-01-01

    Hairy carbon is a new material developed by growing submicron carbon filaments on conventional carbon substrates. Typical substrate materials include carbon black, graphite powder, carbon fibers, and glassy carbon. A catalyst is used to initiate hair growth with carbonaceous gases serving as the carbon source. To study the electrochemical behavior of hairy carbons, cyclic voltammetry (CV) and discharge testing were conducted. In both cases, hairy carbon results surpassed those of the substrate material alone.

  15. The redox status of experimental hemorrhagic shock as measured by cyclic voltammetry.

    Science.gov (United States)

    Mittal, Anubhav; Göke, Friederike; Flint, Richard; Loveday, Benjamin P T; Thompson, Nichola; Delahunt, Brett; Kilmartin, Paul A; Cooper, Garth J S; MacDonald, Julia; Hickey, Anthony; Windsor, John A; Phillips, Anthony R J

    2010-05-01

    Hemorrhagic shock (HS) leads to reactive oxygen species production. However, clinicians do not have access to bedside measurements of the redox status during HS. Cyclic voltammetry (CyV) is a simple electrochemical method of measuring redox status. The aims of this study were to 1) report the first application of cyclic voltammetry to measure the acute changes in serum redox status after HS, 2) to contrast it with another severe systemic disease with a different redox pathology (acute pancreatitis [AP]), and 3) to describe the response of CyV over time in a resolving model of AP. In the acute study, 24 male Wistar rats were randomized into three groups: groups 1 (control), 2 (AP), and 3 (HS). In the time-course study, 28 rats were randomized to a sham-control as well as 6 and 24 h post-AP cohorts, respectively.Cyclic voltammetry was performed using a three-electrode system. In the acute study, the first and second voltammetric peaks increased significantly in HS. In contrast, within the AP group, only the first voltammetric peak showed a significant increase. The first voltammetric peak correlated with plasma protein carbonyls (PCs) and with thiobarbituric acid-reactive substances, whereas the second voltammetric peak correlated positively with plasma protein carbonyls. In the second study, the first voltammetric peak correlated with physiological improvements. Here, we showed that serum CyV could respond to the serum redox change in HS and AP. Cyclic voltammetry warrants evaluation as a potential real-time beside measure of a patient's redox status during shock.

  16. A one-dimensional stochastic approach to the study of cyclic voltammetry with adsorption effects

    OpenAIRE

    Adib J. Samin

    2016-01-01

    In this study, a one-dimensional stochastic model based on the random walk approach is used to simulate cyclic voltammetry. The model takes into account mass transport, kinetics of the redox reactions, adsorption effects and changes in the morphology of the electrode. The model is shown to display the expected behavior. Furthermore, the model shows consistent qualitative agreement with a finite difference solution. This approach allows for an understanding of phenomena on a microscopic level ...

  17. New aspects of the electrochemical-catalytic (EC’) mechanism in square-wave voltammetry

    OpenAIRE

    Gulaboski, Rubin; Mirceski, Valentin

    2015-01-01

    Several new theoretical aspects of the electrocatalytic (regenerative) EC’ mechanism under conditions of square-wave (SWV) and staircase cyclic voltammetry (SCV) are presented. Elaborating the effect of the rate of the catalytic reaction in the diffusion-controlled catalytic mechanism (diffusional EC’ mechanism) and surface catalytic mechanism (surface EC’ mechanism), we refer to several phenomena related to the shift of the position and the half-peak width of the net peak in ...

  18. Glyphosate detection with ammonium nitrate and humic acids as potential interfering substances by pulsed voltammetry technique

    OpenAIRE

    MARTÍNEZ GIL, PABLO; Laguarda Miró, Nicolás; Soto Camino, Juan; Masot Peris, Rafael

    2013-01-01

    Pulsed voltammetry has been used to detect and quantify glyphosate on buffered water in presence of ammonium nitrate and humic substances. Glyphosate is the most widely used herbicide active ingredient in the world. It is a non-selective broad spectrum herbicide but some of its health and environmental effects are still being discussed. Nowadays, glyphosate pollution in water is being monitored but quantification techniques are slow and expensive. Glyphosate wastes are often detected in count...

  19. Dating copper-based archaeological materials using the voltammetry of microparticles

    OpenAIRE

    Doménech-Carbó, Antonio; Capelo, Sofia; Doménech-Carbó, María Teresa

    2014-01-01

    The voltammetry of microparticles, an electrochemical technique providing information on the composition of archaeological materials using an essentially non invasive analysis [1,2] was previously applied for dating lead-based materials [3,4]. It is described the application of this methodology for dating copper-based archaeological materials based on a theoretical model for long term metal corrosion [5]. Dating is based on the measurement of the voltammetric signals of cuprite and tenorite f...

  20. Detection of heavy metals in biological samples through anodic stripping voltammetry

    OpenAIRE

    Buzea, Vlad; Florescu, Monica; Badea, Mihaela

    2012-01-01

    The toxicological aspects due to the presence of heavy metals in biological samples impose to have accurate and rapid methods for their detection. This paper is aimed to review approaches to anodic stripping voltammetry (ASV) determination of several heavy metals (lead, cadmium, copper, mercury, zinc) in biological matrices (blood, urine, saliva, tissue sample). Analytical performances (LOD, data linearity range, sensitivity) of the reviewed methods were presented for several electrochemical ...

  1. Electrode Calibration with a Microfluidic Flow Cell for Fast-scan Cyclic Voltammetry

    OpenAIRE

    Sinkala, Elly; McCutcheon, James E.; Schuck, Matt; Schmidt, Eric; Roitman, Mitchell F.; Eddington, David T.

    2012-01-01

    Fast-scan cyclic voltammetry (FSCV) is a common analytical electrochemistry tool used to measure chemical species. It has recently been adapted for measurement of neurotransmitters such as dopamine in awake and behaving animals (in vivo). Electrode calibration is an essential step in FSCV to relate observed current to concentration of a chemical species. However, existing methods require multiple components, which reduce the ease of calibrations. To this end, a microfluidic flow cell (µFC) wa...

  2. Graphene electroanalysis: inhibitory effects in the stripping voltammetry of cadmium with surfactant free graphene.

    Science.gov (United States)

    Brownson, Dale A C; Lacombe, Alexandre C; Kampouris, Dimitrios K; Banks, Craig E

    2012-01-21

    We explore the use of surfactant free graphene towards the electroanalytical sensing of cadmium(II) ions via anodic stripping voltammetry. In line with literature methodologies, we modify an electrode substrate which exhibits relatively fast electron transfer with commercially available graphene which is free from surfactants. Surprisingly, we find that graphene reduces the analytical performance and hence inhibits the electrochemical detection of cadmium(II) ions, with calibration plots in model aqueous solutions revealing no advantages of employing graphene in this analytical context.

  3. Preparation and Cyclic Voltammetry Characterization Of Cu-dipyridyl Imprinted Polymer

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Polymer capable of specific binding to Cu-dipyridyl complex was prepared by molecular imprinting technology.The binding specificity of the polymer to the template (Cu-dipyridyl complex) was investigated by cyclic voltametric scanning using the carbon paste electrode modified by polymer particles in phosphate buffer solution.Factors that influence rebinding of the imprinted polymer were explored.The result demonstrated that the cyclic voltammetry was an efficient approach to explore interactions between template and imprinted polymers.

  4. Simultaneous identification of lead pigments and binding media in paint samples using voltammetry of microparticles

    OpenAIRE

    DOMENECH CARBO, ANTONIO; Domenech Carbo, Mª Teresa; Mas Barberà, Xavier; Ciarrocci, Julia

    2007-01-01

    Voltammetry of microparticles is applied to the simultaneous determination of lead pigments and binding media in paint samples. The mechanical attachment of a few nanograms of sample to a paraffin-impregnated graphite electrode produced well-defined square wave voltammetric responses for model paint specimens containing lead white, minium and Naples yellow associated with linseed, sunflower and poppy oils and casein, egg, and bovine gelatin. The use of a multiparametric fitting of the electro...

  5. Toward an in situ phosphate sensor in seawater using Square Wave Voltammetry.

    Science.gov (United States)

    Barus, C; Romanytsia, I; Striebig, N; Garçon, V

    2016-11-01

    A Square Wave Voltammetry electrochemical method is proposed to measure phosphate in seawater as pulse techniques offer a higher sensitivity as compared to classical cyclic voltammetry. Chronoamperometry cannot be either adapted for an in situ sensor since this method requires to have controlled convection which will be impossible in a miniaturised sensor. Tests and validation of Square Wave Voltammetry parameters have been performed using an open cell and for the first time with a small volume (<400µL) laboratory prototypes. Two designs of prototypes have been compared. Using high frequency (f=250Hz) allows to obtain a linear behaviour between 0.1 and 1µmolL(-1) with a very low limit of detection of 0.05 µmolL(-1) after 60min of complexation waiting time. In order to obtain a linear regression for a larger concentration range i.e. 0.25-4µmolL(-1), a lower frequency of 2.5Hz is needed. A limit of detection of 0.1µmolL(-1) is obtained in this case after 30min of complexation waiting time for the peak measured at E=0.12V. Changing the position of the molybdenum electrode for the complexation step and moving the detection into another electrochemical cell allow to decrease the reaction time down to 5min.

  6. Development of a cyclic voltammetry method for the detection of Clostridium novyi in black disease.

    Science.gov (United States)

    Liu, L L; Jiang, D N; Xiang, G M; Liu, C; Yu, J C; Pu, X Y

    2014-03-17

    Black disease is an acute disease of sheep and cattle. The pathogen is the obligate anaerobe, Clostridium novyi. Due to difficulties of anaerobic culturing in the country or disaster sites, a simple, rapid, and sensitive method is required. In this study, an electrochemical method, the cyclic voltammetry method, basing on loop-mediated isothermal amplification (LAMP), electrochemical ion bonding (positive dye, methylene blue), was introduced. DNA extracted from C. novyi specimens was amplified through the LAMP reaction. Then the products combined were with methylene blue, which lead to a reduction in the oxidation peak current (ipA) and the reduction peak current (ipC) of the cyclic voltammetry. The changes of ipA/ipC were real-time measured by special designed electrode, so the DNA was quantitatively detected. The results displayed that this electrochemical detection of C. novyi could be completed in 1-2 h with the lowest bacterial concentration of 10(2) colony forming units/mL, and high accuracy (96.5%), sensitivity (96%), and specificity (97%) compared to polymerase chain reation. The cyclic voltammetry method was a simple and fast method, with high sensitivity and high specificity, and has great potential to be a usable molecular tool for fast diagnosis of Black disease.

  7. Alternating current scanning electrochemical microscopy with simultaneous fast-scan cyclic voltammetry.

    Science.gov (United States)

    Koch, Jason A; Baur, Melinda B; Woodall, Erica L; Baur, John E

    2012-11-06

    Fast-scan cyclic voltammetry (FSCV) is combined with alternating current scanning electrochemical microscopy (AC-SECM) for simultaneous measurements of impedance and faradaic current. Scan rates of 10-1000 V s(-1) were used for voltammetry, while a high-frequency (100 kHz), low-amplitude (10 mV rms) sine wave was added to the voltammetric waveform for the ac measurement. Both a lock-in amplifier and an analog circuit were used to measure the amplitude of the resultant ac signal. The effect of the added sine wave on the voltammetry at a carbon fiber electrode was investigated and found to have negligible effect. The combined FSCV and ac measurements were used to provide simultaneous chemical and topographical information about a substrate using a single carbon fiber probe. The technique is demonstrated in living cell culture, where cellular respiration and topography were simultaneously imaged without the addition of a redox mediator. This approach promises to be useful for the topographical and multidimensional chemical imaging of substrates.

  8. Catalytic mechanism in cyclic voltammetry at disc electrodes: an analytical solution.

    Science.gov (United States)

    Molina, Angela; González, Joaquín; Laborda, Eduardo; Wang, Yijun; Compton, Richard G

    2011-08-28

    The theory of cyclic voltammetry at disc electrodes and microelectrodes is developed for a system where the electroactive reactant is regenerated in solution using a catalyst. This catalytic process is of wide importance, not least in chemical sensing, and it can be characterized by the resulting peak current which is always larger than that of a simple electrochemical reaction; in contrast the reverse peak is always relatively diminished in size. From the theoretical point of view, the problem involves a complex physical situation with two-dimensional mass transport and non-uniform surface gradients. Because of this complexity, hitherto the treatment of this problem has been tackled mainly by means of numerical methods and so no analytical expression was available for the transient response of the catalytic mechanism in cyclic voltammetry when disc electrodes, the most popular practical geometry, are used. In this work, this gap is filled by presenting an analytical solution for the application of any sequence of potential pulses and, in particular, for cyclic voltammetry. The induction principle is applied to demonstrate mathematically that the superposition principle applies whatever the geometry of the electrode, which enabled us to obtain an analytical equation valid whatever the electrode size and the kinetics of the catalytic reaction. The theoretical results obtained are applied to the experimental study of the electrocatalytic Fenton reaction, determining the rate constant of the reduction of hydrogen peroxide by iron(II).

  9. Electrochemical characterisation and anodic stripping voltammetry at mesoporous platinum rotating disc electrodes.

    Science.gov (United States)

    Lozano-Sanchez, Pablo; Elliott, Joanne M

    2008-02-01

    Using the technique of liquid crystal templating a rotating disc electrode (RDE) was modified with a high surface area mesoporous platinum film. The surface area of the electrode was characterised by acid voltammetry, and found to be very high (ca. 86 cm(2)). Acid characterisation of the electrode produced distorted voltammograms was interpreted as being due to the extremely large surface area which produced a combination of effects such as localised pH change within the pore environment and also ohmic drop effects. Acid voltammetry in the presence of two different types of surfactant, namely Tween 20 and Triton X-100, suggested antifouling properties associated with the mesoporous deposit. Further analysis of the modified electrode using a redox couple in solution showed typical RDE behaviour although extra capacitive currents were observed due to the large surface area of the electrode. The phenomenon of underpotential deposition was exploited for the purpose of anodic stripping voltammetry and results were compared with data collected for microelectrodes. Underpotential deposition of metal ions at the mesoporous RDE was found to be similar to that at conventional platinum electrodes and mesoporous microelectrodes although the rate of surface coverage was found to be slower at a mesoporous RDE. It was found that a mesoporous RDE forms a suitable system for quantification of silver ions in solution.

  10. Copper-based electrochemical sensor with palladium electrode for cathodic stripping voltammetry of manganese.

    Science.gov (United States)

    Kang, Wenjing; Pei, Xing; Bange, Adam; Haynes, Erin N; Heineman, William R; Papautsky, Ian

    2014-12-16

    In this work, we report on the development of a palladium-based, microfabricated point-of-care electrochemical sensor for the determination of manganese using square wave cathodic stripping voltammetry. Heavy metals require careful monitoring, yet current methods are too complex for a point-of-care system. Voltammetry offers an attractive approach to metal detection on the microscale, but traditional carbon, gold, or platinum electrodes are difficult or expensive to microfabricate, preventing widespread use. Our sensor uses palladium working and auxiliary electrodes and integrates them with a copper-based reference electrode for simple fabrication and compatibility with microfabrication and printed circuit board processing, while maintaining competitive performance in electrochemical detection. Copper electrodes were prepared on glass substrate using a combination of microfabrication procedures followed by electrodeposition of palladium. The disposable sensor system was formed by bonding a poly(dimethylsiloxane) (PDMS) well to the glass substrate. Cathodic stripping voltammetry of manganese using our new disposable palladium-based sensors exhibited 334 nM (18.3 ppb) limit of detection in borate buffer. The sensor was used to demonstrate manganese determination in natural water samples from a pond in Burnet Woods, located in Cincinnati, OH, and the Ohio River.

  11. Study on the adsorptive catalytic voltammetry of aloe-emodin at a carbon paste electrode

    Institute of Scientific and Technical Information of China (English)

    LI; Ju'nan; GAO; Peng; LI; Xiangling; YAN; Zhihong; MAO; Xu

    2005-01-01

    A new catalytic voltammetric method for the determination of anthraqunone medicines at a carbon paste electrode (CPE) was described for the first time. The mechanism of the catalytic reaction was investigated by using linear sweep voltammetry, cyclic voltammetry, constant potential electrolysis and so on. The experiment results indicate that aloe-emodin was efficiently accumulated at a CPE by adsorption. In the following potential scan, aloe-emodin was reduced to homologous anthrahydroquinone compound, then the compound was immediately oxidized to aloe-emodin by the dissolved oxygen, and the aloe-emodin was again reduced at the CPE. As a result, a cyclic catalytic reaction was established. But a reversible redox reaction of aloe-emodin can only be observed at a mercury electrode, no catalytic reaction occurs there. A sensitive catalytic voltammetric peak of aloe-emodin was obtained at about -0.60 V (vs. SCE) in 0.56 mol/L NH3-NH4Cl buffer (pH 8.9). The proposed method was applied to the determination of aloe-emodin in the Radix Rhei with satisfactory results. The determination results were in good agreement with reference values obtained by the HPLC. The adsorptive catalytic voltammetry for the determination of organic compound at CPE, chemically modified electrode and other solid electrodes could be significant in the studies on pharmacology, pharmacodynamics, toxicity of medicine, clinical medicine and biochemistry.

  12. Transcription Factors Efg1 and Bcr1 Regulate Biofilm Formation and Virulence during Candida albicans-Associated Denture Stomatitis.

    Science.gov (United States)

    Yano, Junko; Yu, Alika; Fidel, Paul L; Noverr, Mairi C

    2016-01-01

    Denture stomatitis (DS) is characterized by inflammation of the oral mucosa in direct contact with dentures and affects a significant number of otherwise healthy denture wearers. The disease is caused by Candida albicans, which readily colonizes and form biofilms on denture materials. While evidence for biofilms on abiotic and biotic surfaces initiating Candida infections is accumulating, a role for biofilms in DS remains unclear. Using an established model of DS in immunocompetent animals, the purpose of this study was to determine the role of biofilm formation in mucosal damage during pathogenesis using C. albicans or mutants defective in morphogenesis (efg1-/-) or biofilm formation (bcr1-/-). For in vivo analyses, rats fitted with custom dentures, consisting of fixed and removable parts, were inoculated with wild-type C. albicans, mutants or reconstituted strains and monitored weekly for fungal burden (denture and palate), body weight and tissue damage (LDH) for up to 8 weeks. C. albicans wild-type and reconstituted mutants formed biofilms on dentures and palatal tissues under in vitro, ex vivo and in vivo conditions as indicated by microscopy demonstrating robust biofilm architecture and extracellular matrix (ECM). In contrast, both efg1-/- and bcr1-/- mutants exhibited poor biofilm growth with little to no ECM. In addition, quantification of fungal burden showed reduced colonization throughout the infection period on dentures and palates of rats inoculated with efg1-/-, but not bcr1-/-, compared to controls. Finally, rats inoculated with efg1-/- and bcr1-/- mutants had minimal palatal tissue damage/weight loss while those inoculated with wild-type or reconstituted mutants showed evidence of tissue damage and exhibited stunted weight gain. These data suggest that biofilm formation is associated with tissue damage during DS and that Efg1 and Bcr1, both central regulators of virulence in C. albicans, have pivotal roles in pathogenesis of DS.

  13. Transcriptional and functional analysis of the effects of magnolol: inhibition of autolysis and biofilms in Staphylococcus aureus.

    Directory of Open Access Journals (Sweden)

    Dacheng Wang

    Full Text Available BACKGROUND: The targeting of Staphylococcus aureus biofilm structures are now gaining interest as an alternative strategy for developing new types of antimicrobial agents. Magnolol (MOL shows inhibitory activity against S. aureus biofilms and Triton X-100-induced autolysis in vitro, although there are no data regarding the molecular mechanisms of MOL action in bacteria. METHODOLOGY/PRINCIPAL FINDINGS: The molecular basis of the markedly reduced autolytic phenotype and biofilm inhibition triggered by MOL were explored using transcriptomic analysis, and the transcription of important genes were verified by real-time RT-PCR. The inhibition of autolysis by MOL was evaluated using quantitative bacteriolytic assays and zymographic analysis, and antibiofilm activity assays and confocal laser scanning microscopy were used to elucidate the inhibition of biofilm formation caused by MOL in 20 clinical isolates or standard strains. The reduction in cidA, atl, sle1, and lytN transcript levels following MOL treatment was consistent with the induced expression of their autolytic repressors lrgA, lrgB, arlR, and sarA. MOL generally inhibited or reversed the expression of most of the genes involved in biofilm production. The growth of S. aureus strain ATCC 25923 in the presence of MOL dose-dependently led to decreases in Triton X-100-induced autolysis, extracellular murein hydrolase activity, and the amount of extracellular DNA (eDNA. MOL may impede biofilm formation by reducing the expression of cidA, a murein hydrolase regulator, to inhibit autolysis and eDNA release, or MOL may directly repress biofilm formation. CONCLUSIONS/SIGNIFICANCE: MOL shows in vitro antimicrobial activity against clinical and standard S. aureus strains grown in planktonic and biofilm cultures, suggesting that the structure of MOL may potentially be used as a basis for the development of drugs targeting biofilms.

  14. Transcription Factors Efg1 and Bcr1 Regulate Biofilm Formation and Virulence during Candida albicans-Associated Denture Stomatitis.

    Directory of Open Access Journals (Sweden)

    Junko Yano

    Full Text Available Denture stomatitis (DS is characterized by inflammation of the oral mucosa in direct contact with dentures and affects a significant number of otherwise healthy denture wearers. The disease is caused by Candida albicans, which readily colonizes and form biofilms on denture materials. While evidence for biofilms on abiotic and biotic surfaces initiating Candida infections is accumulating, a role for biofilms in DS remains unclear. Using an established model of DS in immunocompetent animals, the purpose of this study was to determine the role of biofilm formation in mucosal damage during pathogenesis using C. albicans or mutants defective in morphogenesis (efg1-/- or biofilm formation (bcr1-/-. For in vivo analyses, rats fitted with custom dentures, consisting of fixed and removable parts, were inoculated with wild-type C. albicans, mutants or reconstituted strains and monitored weekly for fungal burden (denture and palate, body weight and tissue damage (LDH for up to 8 weeks. C. albicans wild-type and reconstituted mutants formed biofilms on dentures and palatal tissues under in vitro, ex vivo and in vivo conditions as indicated by microscopy demonstrating robust biofilm architecture and extracellular matrix (ECM. In contrast, both efg1-/- and bcr1-/- mutants exhibited poor biofilm growth with little to no ECM. In addition, quantification of fungal burden showed reduced colonization throughout the infection period on dentures and palates of rats inoculated with efg1-/-, but not bcr1-/-, compared to controls. Finally, rats inoculated with efg1-/- and bcr1-/- mutants had minimal palatal tissue damage/weight loss while those inoculated with wild-type or reconstituted mutants showed evidence of tissue damage and exhibited stunted weight gain. These data suggest that biofilm formation is associated with tissue damage during DS and that Efg1 and Bcr1, both central regulators of virulence in C. albicans, have pivotal roles in pathogenesis of DS.

  15. Effect of biofilm formation on the performance of microbial fuel cell for the treatment of palm oil mill effluent.

    Science.gov (United States)

    Baranitharan, E; Khan, Maksudur R; Prasad, D M R; Teo, Wee Fei Aaron; Tan, Geok Yuan Annie; Jose, Rajan

    2015-01-01

    Anode biofilm is a crucial component in microbial fuel cells (MFCs) for electrogenesis. Better knowledge about the biofilm development process on electrode surface is believed to improve MFC performance. In this study, double-chamber microbial fuel cell was operated with diluted POME (initial COD = 1,000 mg L(-1)) and polyacrylonitrile carbon felt was used as electrode. The maximum power density, COD removal efficiency and Coulombic efficiency were found as 22 mW m(-2), 70 and 24 %, respectively. FTIR and TGA analysis confirmed the formation of biofilm on the electrode surface during MFC operation. The impact of anode biofilm on anodic polarization resistance was investigated using electrochemical impedance spectroscopy (EIS) and microbial community changes during MFC operation using denaturing gradient gel electrophoresis (DGGE). The EIS-simulated results showed the reduction of charge transfer resistance (R ct) by 16.9 % after 14 days of operation of the cell, which confirms that the development of the microbial biofilm on the anode decreases the R ct and therefore improves power generation. DGGE analysis showed the variation in the biofilm composition during the biofilm growth until it forms an initial stable microbial community, thereafter the change in the diversity would be less. The power density showed was directly dependent on the biofilm development and increased significantly during the initial biofilm development period. Furthermore, DGGE patterns obtained from 7th and 14th day suggest the presence of less diversity and probable functional redundancy within the anodic communities possibly responsible for the stable MFC performance in changing environmental conditions.

  16. 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

  17. Biofilms in Infections of the Eye

    Directory of Open Access Journals (Sweden)

    Paulo J. M. Bispo

    2015-03-01

    Full Text Available The ability to form biofilms in a variety of environments is a common trait of bacteria, and may represent one of the earliest defenses against predation. Biofilms are multicellular communities usually held together by a polymeric matrix, ranging from capsular material to cell lysate. In a structure that imposes diffusion limits, environmental microgradients arise to which individual bacteria adapt their physiologies, resulting in the gamut of physiological diversity. Additionally, the proximity of cells within the biofilm creates the opportunity for coordinated behaviors through cell–cell communication using diffusible signals, the most well documented being quorum sensing. Biofilms form on abiotic or biotic surfaces, and because of that are associated with a large proportion of human infections. Biofilm formation imposes a limitation on the uses and design of ocular devices, such as intraocular lenses, posterior contact lenses, scleral buckles, conjunctival plugs, lacrimal intubation devices and orbital implants. In the absence of abiotic materials, biofilms have been observed on the capsule, and in the corneal stroma. As the evidence for the involvement of microbial biofilms in many ocular infections has become compelling, developing new strategies to prevent their formation or to eradicate them at the site of infection, has become a priority.

  18. Oral biofilm architecture on natural teeth.

    Science.gov (United States)

    Zijnge, Vincent; van Leeuwen, M Barbara M; Degener, John E; Abbas, Frank; Thurnheer, Thomas; Gmür, Rudolf; Harmsen, Hermie J M

    2010-02-24

    Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and accessibility. Despite descriptions of initial plaque formation on the tooth surface, studies on mature plaque and plaque structure below the gum are limited to landmark studies from the 1970s, without appreciating the breadth of microbial diversity in the plaque. We used fluorescent in situ hybridization to localize in vivo the most abundant species from different phyla and species associated with periodontitis on seven embedded teeth obtained from four different subjects. The data showed convincingly the dominance of Actinomyces sp., Tannerella forsythia, Fusobacterium nucleatum, Spirochaetes, and Synergistetes in subgingival plaque. The latter proved to be new with a possibly important role in host-pathogen interaction due to its localization in close proximity to immune cells. The present study identified for the first time in vivo that Lactobacillus sp. are the central cells of bacterial aggregates in subgingival plaque, and that Streptococcus sp. and the yeast Candida albicans form corncob structures in supragingival plaque. Finally, periodontal pathogens colonize already formed biofilms and form microcolonies therein. These in vivo observations on oral biofilms provide a clear vision on biofilm architecture and the spatial distribution of predominant species.

  19. Crenarchaeal biofilm formation under extreme conditions.

    Directory of Open Access Journals (Sweden)

    Andrea Koerdt

    Full Text Available BACKGROUND: Biofilm formation has been studied in much detail for a variety of bacterial species, as it plays a major role in the pathogenicity of bacteria. However, only limited information is available for the development of archaeal communities that are frequently found in many natural environments. METHODOLOGY: We have analyzed biofilm formation in three closely related hyperthermophilic crenarchaeotes: Sulfolobus acidocaldarius, S. solfataricus and S. tokodaii. We established a microtitre plate assay adapted to high temperatures to determine how pH and temperature influence biofilm formation in these organisms. Biofilm analysis by confocal laser scanning microscopy demonstrated that the three strains form very different communities ranging from simple carpet-like structures in S. solfataricus to high density tower-like structures in S. acidocaldarius in static systems. Lectin staining indicated that all three strains produced extracellular polysaccharides containing glucose, galactose, mannose and N-acetylglucosamine once biofilm formation was initiated. While flagella mutants had no phenotype in two days old static biofilms of S. solfataricus, a UV-induced pili deletion mutant showed decreased attachment of cells. CONCLUSION: The study gives first insights into formation and development of crenarchaeal biofilms in extreme environments.

  20. Iron and Acinetobacter baumannii Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Valentina Gentile

    2014-08-01

    Full Text Available Acinetobacter baumannii is an emerging nosocomial pathogen, responsible for infection outbreaks worldwide. The pathogenicity of this bacterium is mainly due to its multidrug-resistance and ability to form biofilm on abiotic surfaces, which facilitate long-term persistence in the hospital setting. Given the crucial role of iron in A. baumannii nutrition and pathogenicity, iron metabolism has been considered as a possible target for chelation-based antibacterial chemotherapy. In this study, we investigated the effect of iron restriction on A. baumannii growth and biofilm formation using different iron chelators and culture conditions. We report substantial inter-strain variability and growth medium-dependence for biofilm formation by A. baumannii isolates from veterinary and clinical sources. Neither planktonic nor biofilm growth of A. baumannii was affected by exogenous chelators. Biofilm formation was either stimulated by iron or not responsive to iron in the majority of isolates tested, indicating that iron starvation is not sensed as an overall biofilm-inducing stimulus by A. baumannii. The impressive iron withholding capacity of this bacterium should be taken into account for future development of chelation-based antimicrobial and anti-biofilm therapies.

  1. Alternating Current Influences Anaerobic Electroactive Biofilm Activity.

    Science.gov (United States)

    Wang, Xin; Zhou, Lean; Lu, Lu; Lobo, Fernanda Leite; Li, Nan; Wang, Heming; Park, Jaedo; Ren, Zhiyong Jason

    2016-09-06

    Alternating current (AC) is known to inactivate microbial growth in suspension, but how AC influences anaerobic biofilm activities has not been systematically investigated. Using a Geobacter dominated anaerobic biofilm growing on the electrodes of microbial electrochemical reactors, we found that high frequency AC ranging from 1 MHz to 1 kHz (amplitude of 5 V, 30 min) showed only temporary inhibition to the biofilm activity. However, lower frequency (100 Hz, 1.2 or 5 V) treatment led to 47 ± 19% permanent decrease in limiting current on the same biofilm, which is attributed to the action of electrohydrodynamic force that caused biofilm damage and loss of intercellular electron transfer network. Confocal microscopy images show such inactivation mainly occurred at the interface between the biofilm and the electrode. Reducing the frequency further to 1 Hz led to water electrolysis, which generated gas bubbles that flushed all attached cells out of the electrode. These findings provide new references on understanding and regulating biofilm growth, which has broader implications in biofouling control, anaerobic waste treatment, energy and product recovery, and general understanding of microbial ecology and physiology.

  2. Oral biofilm architecture on natural teeth.

    Directory of Open Access Journals (Sweden)

    Vincent Zijnge

    Full Text Available Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and accessibility. Despite descriptions of initial plaque formation on the tooth surface, studies on mature plaque and plaque structure below the gum are limited to landmark studies from the 1970s, without appreciating the breadth of microbial diversity in the plaque. We used fluorescent in situ hybridization to localize in vivo the most abundant species from different phyla and species associated with periodontitis on seven embedded teeth obtained from four different subjects. The data showed convincingly the dominance of Actinomyces sp., Tannerella forsythia, Fusobacterium nucleatum, Spirochaetes, and Synergistetes in subgingival plaque. The latter proved to be new with a possibly important role in host-pathogen interaction due to its localization in close proximity to immune cells. The present study identified for the first time in vivo that Lactobacillus sp. are the central cells of bacterial aggregates in subgingival plaque, and that Streptococcus sp. and the yeast Candida albicans form corncob structures in supragingival plaque. Finally, periodontal pathogens colonize already formed biofilms and form microcolonies therein. These in vivo observations on oral biofilms provide a clear vision on biofilm architecture and the spatial distribution of predominant species.

  3. Aging biofilm from a full-scale moving bed biofilm reactor: characterization and enzymatic treatment study.

    Science.gov (United States)

    Huang, Hui; Ren, Hongqiang; Ding, Lili; Geng, Jinju; Xu, Ke; Zhang, Yan

    2014-02-01

    Effective removal of aging biofilm deserves to receive more attention. This study aimed to characterized aging biofilm from a full-scale moving bed biofilm reactor treating pharmaceutical wastewater and evaluate the hydrolysis effects of biofilm by different enzymatic treatments. Results from FTIR and biochemical composition analyses showed that it was a predominately organic-based biofilm with the ratio of total protein (PN) to polysaccharide (PS) of 20.17. A reticular structure of extracellular polymeric matrix (EPM) with filamentous bacteria as the skeleton was observed on the basal layer through SEM-EDS test. Among the four commercial proteases and amylases from Genencor®, proteases were shown to have better performances than amylases either on the removal of MLSS and PN/MLSS or on DOC (i.e., dissolved organic carbon)/MLSS raising of biofilm pellets. Difference of dynamic fluorescence characteristics of dissolved organic matters after treated by the two proteases indicated distinguishing mechanisms of the treating process.

  4. Treatment of Oral Multispecies Biofilms by an Anti-Biofilm Peptide.

    Directory of Open Access Journals (Sweden)

    Zhejun Wang

    Full Text Available Human oral biofilms are multispecies microbial communities that exhibit high resistance to antimicrobial agents. Dental plaque gives rise to highly prevalent and costly biofilm-related oral infections, which lead to caries or other types of oral infections. We investigated the ability of the recently identified anti-biofilm peptide 1018 to induce killing of bacterial cells present within oral multispecies biofilms. At 10 μg/ml (6.5 μM, peptide 1018 was able to significantly (p50% of the biofilm being killed and >35% being dispersed in only 3 minutes. Peptide 1018 may potentially be used by itself or in combination with CHX as a non-toxic and effective anti-biofilm agent for plaque disinfection in clinical dentistry.

  5. Enhanced Biofilm Formation and Increased Resistance to Antimicrobial Agents and Bacterial Invasion Are Caused by Synergistic Interactions in Multispecies Biofilms

    DEFF Research Database (Denmark)

    Burmølle, Mette; Webb, J.S.; Rao, D.

    2006-01-01

    Most biofilms in their natural environments are likely to consist of consortia of species that influence each other in synergistic and antagonistic manners. However, few reports specifically address interactions within multispecies biofilms. In this study, 17 epiphytic bacterial strains, isolated...... specific interactions. In summary, our data strongly indicate that synergistic effects promote biofilm biomass and resistance of the biofilm to antimicrobial agents and bacterial invasion in multispecies biofilms....

  6. Modulation of gut mucosal biofilms.

    Science.gov (United States)

    Kleessen, Brigitta; Blaut, Michael

    2005-04-01

    Non-digestible inulin-type fructans, such as oligofructose and high-molecular-weight inulin, have been shown to have the ability to alter the intestinal microbiota composition in such a way that members of the microbial community, generally considered as health-promoting, are stimulated. Bifidobacteria and lactobacilli are the most frequently targeted organisms. Less information exists on effects of inulin-type fructans on the composition, metabolism and health-related significance of bacteria at or near the mucosa surface or in the mucus layer forming mucosa-associated biofilms. Using rats inoculated with a human faecal flora as an experimental model we have found that inulin-type fructans in the diet modulated the gut microbiota by stimulation of mucosa-associated bifidobacteria as well as by partial reduction of pathogenic Salmonella enterica subsp. enterica serovar Typhimurium and thereby benefit health. In addition to changes in mucosal biofilms, inulin-type fructans also induced changes in the colonic mucosa stimulating proliferation in the crypts, increasing the release of mucins, and altering the profile of mucin components in the goblet cells and epithelial mucus layer. These results indicate that inulin-type fructans may stabilise the gut mucosal barrier. Dietary supplementation with these prebiotics could offer a new approach to supporting the barrier function of the mucosa.

  7. Examining the complex regulation and drug-induced plasticity of dopamine release and uptake using voltammetry in brain slices.

    Science.gov (United States)

    Ferris, Mark J; Calipari, Erin S; Yorgason, Jordan T; Jones, Sara R

    2013-05-15

    Fast scan cyclic voltammetry in brain slices (slice voltammetry) has been used over the last several decades to increase substantially our understanding of the complex local regulation of dopamine release and uptake in the striatum. This technique is routinely used for the study of changes that occur in the dopamine system associated with various disease states and pharmacological treatments, and to study mechanisms of local circuitry regulation of dopamine terminal function. In the context of this Review, we compare the relative advantages of voltammetry using striatal slice preparations versus in vivo preparations, and highlight recent advances in our understanding of dopamine release and uptake in the striatum specifically from studies that use slice voltammetry in drug-naïve animals and animals with a history of psychostimulant self-administration.

  8. Staining of wool using the reaction products of ABTS oxidation by laccase: synergetic effects of ultrasound and cyclic voltammetry.

    Science.gov (United States)

    Munteanu, Florentina-Daniela; Basto, Carlos; Gübitz, Georg M; Cavaco-Paulo, Artur

    2007-03-01

    The effects of ultrasound on 2,2'-Azinobis(3-ethylbenzothiazoline-6-sulfonate) enzymatic oxidation by laccase (Trametes villosa) has been studied by means of cyclic voltammetry. The reaction was allowed to proceed in the presence of a piece of wool and the coloration depth of the wool fabric was measured by means of K/S. It was observed that cyclic voltammetry is influenced the dyeing process and higher K/S values were obtained when the cyclic voltammetry was combined with the ultrasonic irradiation. Moreover, the K/S value is the sum of the values obtained when the wool staining is done in just the presence of cyclic voltammetry or in just the presence of ultrasound. The results obtained on the indigo carmine decolourization gives information on the importance of controlling the amount of ABTS(+) formed during the ultrasonication process.

  9. Implications of Biofilm Formation on Urological Devices

    Science.gov (United States)

    Cadieux, Peter A.; Wignall, Geoffrey R.; Carriveau, Rupp; Denstedt, John D.

    2008-09-01

    Despite millions of dollars and several decades of research targeted at their prevention and eradication, biofilm-associated infections remain the major cause of urological device failure. Numerous strategies have been aimed at improving device design, biomaterial composition, surface properties and drug delivery, but have been largely circumvented by microbes and their plethora of attachment, host evasion, antimicrobial resistance, and dissemination strategies. This is not entirely surprising since natural biofilm formation has been going on for millions of years and remains a major part of microorganism survival and evolution. Thus, the fact that biofilms develop on and in the biomaterials and tissues of humans is really an extension of this natural tendency and greatly explains why they are so difficult for us to combat. Firstly, biofilm structure and composition inherently provide a protective environment for microorganisms, shielding them from the shear stress of urine flow, immune cell attack and some antimicrobials. Secondly, many biofilm organisms enter a metabolically dormant state that renders them tolerant to those antibiotics and host factors able to penetrate the biofilm matrix. Lastly, the majority of organisms that cause biofilm-associated urinary tract infections originate from our own oral cavity, skin, gastrointestinal and urogenital tracts and therefore have already adapted to many of our host defenses. Ultimately, while biofilms continue to hold an advantage with respect to recurrent infections and biomaterial usage within the urinary tract, significant progress has been made in understanding these dynamic microbial communities and novel approaches offer promise for their prevention and eradication. These include novel device designs, antimicrobials, anti-adhesive coatings, biodegradable polymers and biofilm-disrupting compounds and therapies.

  10. Laser Microbial Killing and Biofilm Disruption

    Science.gov (United States)

    Krespi, Yosef P.; Kizhner, Victor

    2009-06-01

    Objectives: To analyze the ability of NIR lasers to reduce bacterial load and demonstrate the capability of fiber-based Q-switched Nd:YAG laser disrupting biofilm. Study Design: NIR diode laser was tested in vitro and in vivo using pathogenic microorganisms (S. aureus, S. pneumoniae, P. aeruginosa). In addition biofilms were grown from clinical Pseudomonas isolates and placed in culture plates, screws, tympanostomy tubes and PET sutures. Methods: In the animal experiments acute rhinosinusitis model was created by packing the rabbit nose with bacteria soaked solution. The nasal pack was removed in two days and nose was exposed to laser irradiation. A 940 nm diode laser with fiber diffuser was used. Nasal cultures were obtained before and after the laser treatments. Animals were sacrificed fifteen days following laser treatment and bacteriologic/histologic results analyzed. Q-switched Nd:YAG laser generated shockwave pulses were delivered on biofilm using special probes over culture plates, screws, tubes, and PET sutures for the biofilm experiments. Results: Average of two log bacteria reduction was achieved with NIR laser compared to controls. Histologic studies demonstrated preservation of tissue integrity without significant damage to mucosa. Biofilms were imaged before, during and after treatment using a confocal microscope. During laser-generated shockwave application, biofilm was initially seen to oscillate and eventually break off. Large and small pieces of biofilm were totally and instantly removed from the surface to which they were attached in seconds. Conclusions: Significant bacterial reduction was achieved with NIR laser therapy in this experimental in vitro and animal study. In addition we disrupted Pseudomonas aeruginosa biofilms using Q-switched Nd:YAG laser and special probes generating plasma and shockwave. This new and innovative method of bacteria killing and biofilm disruption without injuring host tissue may have clinical application in the

  11. Prevention of biofilm formation and removal of existing biofilms by extracellular DNases of Campylobacter jejuni.

    Science.gov (United States)

    Brown, Helen L; Reuter, Mark; Hanman, Kate; Betts, Roy P; van Vliet, Arnoud H M

    2015-01-01

    The fastidious nature of the foodborne bacterial pathogen Campylobacter jejuni contrasts with its ability to survive in the food chain. The formation of biofilms, or the integration into existing biofilms by C. jejuni, is thought to contribute to food chain survival. As extracellular DNA (eDNA) has previously been proposed to play a role in C. jejuni biofilms, we have investigated the role of extracellular DNases (eDNases) produced by C. jejuni in biofilm formation. A search of 2791 C. jejuni genomes highlighted that almost half of C. jejuni genomes contains at least one eDNase gene, but only a minority of isolates contains two or three of these eDNase genes, such as C. jejuni strain RM1221 which contains the cje0256, cje0566 and cje1441 eDNase genes. Strain RM1221 did not form biofilms, whereas the eDNase-negative strains NCTC 11168 and 81116 did. Incubation of pre-formed biofilms of NCTC 11168 with live C. jejuni RM1221 or with spent medium from a RM1221 culture resulted in removal of the biofilm. Inactivation of the cje1441 eDNase gene in strain RM1221 restored biofilm formation, and made the mutant unable to degrade biofilms of strain NCTC 11168. Finally, C. jejuni strain RM1221 was able to degrade genomic DNA from C. jejuni NCTC 11168, 81116 and RM1221, whereas strain NCTC 11168 and the RM1221 cje1441 mutant were unable to do so. This was mirrored by an absence of eDNA in overnight cultures of C. jejuni RM1221. This suggests that the activity of eDNases in C. jejuni affects biofilm formation and is not conducive to a biofilm lifestyle. These eDNases do however have a potential role in controlling biofilm formation by C. jejuni strains in food chain relevant environments.

  12. A novel approach for harnessing biofilm communities in moving bed biofilm reactors for industrial wastewater treatment

    OpenAIRE

    Joe A Lemire; Demeter, Marc A.; Iain George; Howard Ceri; Turner, Raymond J.

    2015-01-01

    Moving bed biofilm reactors (MBBRs) are an effective biotechnology for treating industrial wastewater. Biomass retention on moving bed biofilm reactor (MBBR) carriers (biofilm support materials), allows for the ease-of-operation and high treatment capacity of MBBR systems. Optimization of MBBR systems has largely focused on aspects of carrier design, while little attention has been paid to enhancing strategies for harnessing microbial biomass. Previously, our research group demonstrated that ...

  13. Ferrocene-Boronic Acid-Fructose Binding Based on Dual-Plate Generator-Collector Voltammetry and Square-Wave Voltammetry.

    Science.gov (United States)

    Li, Meng; Xu, Su-Ying; Gross, Andrew J; Hammond, Jules L; Estrela, Pedro; Weber, James; Lacina, Karel; James, Tony D; Marken, Frank

    2015-06-10

    The interaction of ferrocene-boronic acid with fructose is investigated in aqueous 0.1 m phosphate buffer at pH 7, 8 and 9. Two voltammetric methods, based on 1) a dual-plate generator-collector micro-trench electrode (steady state) and 2) a square-wave voltammetry (transient) method, are applied and compared in terms of mechanistic resolution. A combination of experimental data is employed to obtain new insights into the binding rates and the cumulative binding constants for both the reduced ferrocene-boronic acid (pH dependent and weakly binding) and for the oxidised ferrocene-boronic acid (pH independent and strongly binding).

  14. Screening selectively harnessed environmental microbial communities for biodegradation of polycyclic aromatic hydrocarbons in moving bed biofilm reactors.

    Science.gov (United States)

    Demeter, Marc A; Lemire, Joseph A; Mercer, Sean M; Turner, Raymond J

    2017-03-01

    Bacteria are often found tolerating polluted environments. Such bacteria may be exploited to bioremediate contaminants in controlled ex situ reactor systems. One potential strategic goal of such systems is to harness microbes directly from the environment such that they exhibit the capacity to markedly degrade organic pollutants of interest. Here, the use of biofilm cultivation techniques to inoculate and activate moving bed biofilm reactor (MBBR) systems for the degradation of polycyclic aromatic hydrocarbons (PAHs) was explored. Biofilms were cultivated from 4 different hydrocarbon contaminated sites using a minimal medium spiked with the 16 EPA identified PAHs. Overall, all 4 inoculant sources resulted in biofilm communities capable of tolerating the presence of PAHs, but only 2 of these exhibited enhanced PAH catabolic gene prevalence coupled with significant degradation of select PAH compounds. Comparisons between inoculant sources highlighted the dependence of this method on appropriate inoculant screening and biostimulation efforts.

  15. Biofilm formation in a hot water system

    DEFF Research Database (Denmark)

    Bagh, L.K.; Albrechtsen, Hans-Jørgen; Arvin, Erik

    2002-01-01

    The biofilm formation rate was measured in situ in a hot water system in an apartment building by specially designed sampling equipment, and the net growth of the suspended bacteria was measured by incubation of water samples with the indigeneous bacteria. The biofilm formation rate reached......, in the sludge, or in the water from the distribution system was negligible. This indicated that bacterial growth took place on the inner surfaces in the hot water system and biofilm formation and detachment of bacteria could account for most of the suspended bacteria actually measured in hot water. Therefore...

  16. Anaerobic granular sludge and biofilm reactors

    DEFF Research Database (Denmark)

    Skiadas, Ioannis V.; Gavala, Hariklia N.; Schmidt, Jens Ejbye

    2003-01-01

    by the immobilization of the biomass, which forms static biofilms, particle-supported biofilms, or granules depending on the reactor's operational conditions. The advantages of the high-rate anaerobic digestion over the conventional aerobic wastewater treatment methods has created a clear trend for the change......-rate anaerobic treatment systems based on anaerobic granular sludge and biofilm are described in this chapter. Emphasis is given to a) the Up-flow Anaerobic Sludge Blanket (UASB) systems, b) the main characteristics of the anaerobic granular sludge, and c) the factors that control the granulation process...

  17. Quorum sensing inhibitors disable bacterial biofilms

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Tolker-Nielsen, Tim; Givskov, Michael

    2011-01-01

    It is now evident that bacteria assume the biofilm mode of growth during chronic infections. The important hallmarks of biofilm infections are development of local inflammations, extreme tolerance to the action of conventional antimicrobial agents and an almost infinite capacity to evade the host...... defence systems in particular innate immunity. In the biofilm mode, bacteria use cell to cell communication termed quorum-sensing (QS) to coordinate expression of virulence, tolerance towards a number of antimicrobial agents and shielding against the host defence system. Chemical biology approaches may...

  18. UV-Induced prevention of biofilm formation inside medical tubes and catheters

    DEFF Research Database (Denmark)

    Pedersen, Jens Kristian Mølgaard; Nielsen, Kristian; Bang, Ole

    2014-01-01

    risk connected withthe use of medical tubes and catheters is the direct cause of more than 60% of all infections acquired inEuropean hospitals. Once formed, the biofilm is generally very tough to suppress by either the body’simmunity system or by use of antibiotics, which may even favor the population...... of multi resistant bacteriacultures. Prevention of biofilm formation inside the tube or catheter, without risk of developing multiresistance, may be achieved by creating a UV-exposed environment in the interior. This may be realized bytransforming the tube itself into an optical waveguide supporting UV...

  19. A personal history of research on microbial biofilms and biofilm infections.

    Science.gov (United States)

    Høiby, Niels

    2014-04-01

    The observation of aggregated microorganisms surrounded by a self-produced matrix adhering to surfaces or located in tissues or secretions is as old as microbiology, with both Leeuwenhoek and Pasteur describing the phenomenon. In environmental and technical microbiology, biofilms were already shown 80-90 years ago to be important for biofouling on submerged surfaces, e.g. ships. The concept of biofilm infections and their importance in medicine is, however, biofilm was introduced into medicine in 1985 by Costerton. In the following decades, it became obvious that biofilm infections are widespread in medicine, and their importance is now generally accepted.

  20. Successional development of biofilms in moving bed biofilm reactor (MBBR) systems treating municipal wastewater.

    Science.gov (United States)

    Biswas, Kristi; Taylor, Michael W; Turner, Susan J

    2014-02-01

    Biofilm-based technologies, such as moving bed biofilm reactor (MBBR) systems, are widely used to treat wastewater. Biofilm development is important for MBBR systems as much of the microbial biomass is retained within reactors as biofilm on suspended carriers. Little is known about this process of biofilm development and the microorganisms upon which MBBRs rely. We documented successional changes in microbial communities as biofilms established in two full-scale MBBR systems treating municipal wastewater over two seasons. 16S rRNA gene-targeted pyrosequencing and clone libraries were used to describe microbial communities. These data indicate a successional process that commences with the establishment of an aerobic community dominated by Gammaproteobacteria (up to 52 % of sequences). Over time, this community shifts towards dominance by putatively anaerobic organisms including Deltaproteobacteria and Clostridiales. Significant differences were observed between the two wastewater treatment plants (WWTPs), mostly due to a large number of sequences (up to 55 %) representing Epsilonproteobacteria (mostly Arcobacter) at one site. Archaea in young biofilms included several lineages of Euryarchaeota and Crenarchaeota. In contrast, the mature biofilm consisted entirely of Methanosarcinaceae (Euryarchaeota). This study provides new insights into the community structure of developing biofilms at full-scale WWTPs and provides the basis for optimizing MBBR start-up and operational parameters.

  1. Fungal Biofilms: In Vivo Models for Discovery of Anti-Biofilm Drugs.

    Science.gov (United States)

    Nett, Jeniel E; Andes, David R

    2015-06-01

    During infection, fungi frequently transition to a biofilm lifestyle, proliferating as communities of surface-adherent aggregates of cells. Phenotypically, cells in a biofilm are distinct from free-floating cells. Their high tolerance of antifungals and ability to withstand host defenses are two characteristics that foster resilience. Biofilm infections are particularly difficult to eradicate, and most available antifungals have minimal activity. Therefore, the discovery of novel compounds and innovative strategies to treat fungal biofilms is of great interest. Although many fungi have been observed to form biofilms, the most well-studied is Candida albicans. Animal models have been developed to simulate common Candida device-associated infections, including those involving vascular catheters, dentures, urinary catheters, and subcutaneous implants. Models have also reproduced the most common mucosal biofilm infections: oropharyngeal and vaginal candidiasis. These models incorporate the anatomical site, immune components, and fluid dynamics of clinical niches and have been instrumental in the study of drug resistance and investigation of novel therapies. This chapter describes the significance of fungal biofilm infections, the animal models developed for biofilm study, and how these models have contributed to the development of new strategies for the eradication of fungal biofilm infections.

  2. IMPACTS OF BIOFILM FORMATION ON CELLULOSE FERMENTATION

    Energy Technology Data Exchange (ETDEWEB)

    Leschine, Susan

    2009-10-31

    This project addressed four major areas of investigation: i) characterization of formation of Cellulomonas uda biofilms on cellulose; ii) characterization of Clostridium phytofermentans biofilm development; colonization of cellulose and its regulation; iii) characterization of Thermobifida fusca biofilm development; colonization of cellulose and its regulation; and iii) description of the architecture of mature C. uda, C. phytofermentans, and T. fusca biofilms. This research is aimed at advancing understanding of biofilm formation and other complex processes involved in the degradation of the abundant cellulosic biomass, and the biology of the microbes involved. Information obtained from these studies is invaluable in the development of practical applications, such as the single-step bioconversion of cellulose-containing residues to fuels and other bioproducts. Our results have clearly shown that cellulose-decomposing microbes rapidly colonize cellulose and form complex structures typical of biofilms. Furthermore, our observations suggest that, as cells multiply on nutritive surfaces during biofilms formation, dramatic cell morphological changes occur. We speculated that morphological changes, which involve a transition from rod-shaped cells to more rounded forms, might be more apparent in a filamentous microbe. In order to test this hypothesis, we included in our research a study of biofilm formation by T. fusca, a thermophilic cellulolytic actinomycete commonly found in compost. The cellulase system of T. fusca has been extensively detailed through the work of David Wilson and colleagues at Cornell, and also, genome sequence of a T. fusca strain has been determine by the DOE Joint Genome Institute. Thus, T. fusca is an excellent subject for studies of biofilm development and its potential impacts on cellulose degradation. We also completed a study of the chitinase system of C. uda. This work provided essential background information for understanding how C. uda

  3. Diversity of Salmonella in biofilms and water in a headwater ecosystem.

    Science.gov (United States)

    Sha, Qiong; Forstner, Michael R J; Hahn, Dittmar

    2013-03-01

    The diversity of Salmonella was analysed in biofilm and water samples from the spring and slough arms of Spring Lake, the headwaters of the San Marcos River, Texas, during the drought of 2011, with only one potential run-off event at the beginning of the study. Salmonellae were detected in semiselective enrichment cultures by end-point PCR during the entire sampling period of (11 sampling events during 2 months). From the spring arm site, 73% of the biofilms and 41% of the water samples were positive for salmonellae, while only 9% of the biofilms and 23% of the water samples were positive from the slough arm site. Salmonellae could be isolated from all positive samples, with higher diversity in biofilms compared with water samples and more strains obtained from the spring arm than from the slough arm. Differences between sites were generally caused by less frequently detected isolates, while the majority of isolates that were present in both biofilms and water from both sites was represented by three strains only. Quantification attempts by quantitative PCR directly in samples without prior enrichment did not result in a reliable detection of salmonellae, suggesting that numbers in all samples were below the detection limit.

  4. Synergistic effect of polyaniline coverage and surface microstructure on the inhibition of Pseudomonas aeruginosa biofilm formation.

    Science.gov (United States)

    Gallarato, L A; Mulko, L E; Dardanelli, M S; Barbero, C A; Acevedo, D F; Yslas, E I

    2017-02-01

    Biofilm Formation is a survival strategy for microorganisms to adapt to their environment. Microbial cells in biofilm become tolerant and resistant to antibiotics and immune responses, increasing the difficulties for the clinical treatment of microbial infections. The surface chemistry and the micro/nano-topography of solid interfaces play a major role in mediating microorganism activity and adhesion. The effect of the surface chemical composition and topography on the adhesion and viability of Pseudomonas aeruginosa was studied. Polymeric (polyethylene terephthalate) surfaces were covered with a conducting polymer (polyaniline, PANI) film by in-situ polymerization and microstructured by Direct Laser Interference Patterning (DLIP). The viability of Pseudomonas aeruginosa on the different surfaces was investigated. The physicochemical properties of the surfaces were characterized by water contact angle measurements, scanning electron microscopy and atomic force microscopy. Bacterial biofilms were imaged by atomic force and scanning electron microscopies. The bacterial viability decreased on PANI compared with the substrate (polyethylene terephthalate) and it decreased even more upon micro-structuring the PANI films. In addition, the biofilm reduction could be improved using polymers with different chemical composition and/or the same polymer with different topographies. Both methods presented diminish the bacterial attachment and biofilm formation. These findings present a high impact related to materials for biomedical engineer applications regarding medical devices, as prostheses or catheters.

  5. Influence of incubation temperature on biofilm formation and corrosion of carbon steel by Serratia marcescens

    Science.gov (United States)

    Harimawan, Ardiyan; Devianto, Hary; Kurniawan, Ignatius Chandra; Utomo, Josephine Christine

    2017-01-01

    Microbial induced corrosion (MIC) or biocorrosion is one type of corrosion, directly or indirectly influenced by microbial activities, by forming biofilm and adhering on the metal surface. When forming biofilm, the microorganisms can produce extracellular products which influence the cathodic and anodic reactions on metal surfaces. This will result in electrochemical changes in the interface between the biofilm and the metal surface, leading to corrosion and deterioration of the metal. MIC might be caused by various types of microorganism which leads to different corrosion mechanism and reaction kinetics. Furthermore, this process will also be influenced by various environmental conditions, such as pH and temperature. This research is aimed to determine the effect of incubation temperature on corrosion of carbon steel caused by Serratia marcescens in a mixture solution of synthetic seawater with Luria Bertani medium with a ratio of 4:1. The incubation was performed for 19 days with incubation temperature of 30, 37, and 50°C. The analyses of biofilm were conducted by total plate count (TPC), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Biofilm was found to be evenly growth on the surface and increasing with increasing incubation temperature. It consists of functional group of alcohol, alkane, amine, nitro, sulfate, carboxylic acid, and polysulfide. The analyses of the corrosion were conducted by gravimetric and X-ray diffraction (XRD). Higher incubation temperature was found to increase the corrosion rate. However, the corrosion products were not detected by XRD analysis.

  6. Biofilm formation by Streptococcus agalactiae: influence of environmental conditions and implicated virulence factors.

    Science.gov (United States)

    Rosini, Roberto; Margarit, Immaculada

    2015-01-01

    Streptococcus agalactiae (Group B Streptococcus, GBS) is an important human pathogen that colonizes the urogenital and/or the lower gastro-intestinal tract of up to 40% of healthy women of reproductive age and is a leading cause of sepsis and meningitis in the neonates. GBS can also infect the elderly and immuno-compromised adults, and is responsible for mastitis in bovines. Like other Gram-positive bacteria, GBS can form biofilm-like three-dimensional structures that could enhance its ability to colonize and persist in the host. Biofilm formation by GBS has been investigated in vitro and appears tightly controlled by environmental conditions. Several adhesins have been shown to play a role in the formation of GBS biofilm-like structures, among which are the protein components of pili protruding outside the bacterial surface. Remarkably, antibodies directed against pilus proteins can prevent the formation of biofilms. The implications of biofilm formation in the context of GBS asymptomatic colonization and dissemination to cause invasive disease remain to be investigated in detail.

  7. Biofilm formation by Streptococcus agalactiae: influence of environmental conditions and implicated virulence factors

    Directory of Open Access Journals (Sweden)

    Imma eMargarit

    2015-02-01

    Full Text Available Streptococcus agalactiae (Group B Streptococcus, GBS is an important human pathogen that colonizes the urogenital and/or the lower gastro-intestinal tract of up to 40% of healthy women of reproductive age and is a leading cause of sepsis and meningitis in the neonates. GBS can also infect the elderly and immuno-compromised adults, and is responsible for mastitis in bovines. Like other Gram-positive bacteria, GBS can form biofilm-like three-dimensional structures that could enhance its ability to colonize and persist in the host. Biofilm formation by GBS has been investigated in vitro and appears tightly controlled by environmental conditions. Several adhesins have been shown to play a role in the formation of GBS biofilm-like structures, among which are the protein components of pili protruding outside the bacterial surface. Remarkably, antibodies directed against pilus proteins can prevent the formation of biofilms. The implications of biofilm formation in the context of GBS asymptomatic colonization and dissemination to cause invasive disease remain to be investigated in detail.

  8. Antifungal Activity of 14-Helical β-Peptides against Planktonic Cells and Biofilms of Candida Species

    Directory of Open Access Journals (Sweden)

    Namrata Raman

    2015-08-01

    Full Text Available Candida albicans is the most prevalent cause of fungal infections and treatment is further complicated by the formation of drug resistant biofilms, often on the surfaces of implanted medical devices. In recent years, the incidence of fungal infections by other pathogenic Candida species such as C. glabrata, C. parapsilosis and C. tropicalis has increased. Amphiphilic, helical β-peptide structural mimetics of natural antimicrobial α-peptides have been shown to exhibit specific planktonic antifungal and anti-biofilm formation activity against C. albicans in vitro. Here, we demonstrate that β-peptides are also active against clinically isolated and drug resistant strains of C. albicans and against other opportunistic Candida spp. Different Candida species were susceptible to β-peptides to varying degrees, with C. tropicalis being the most and C. glabrata being the least susceptible. β-peptide hydrophobicity directly correlated with antifungal activity against all the Candida clinical strains and species tested. While β-peptides were largely ineffective at disrupting existing Candida biofilms, hydrophobic β-peptides were able to prevent the formation of C. albicans, C. glabrata, C. parapsilosis and C. tropicalis biofilms. The broad-spectrum antifungal activity of β-peptides against planktonic cells and in preventing biofilm formation suggests the promise of this class of molecules as therapeutics.

  9. Biofilm growth in porous media: experiments, computational modeling at the porescale, and upscaling

    CERN Document Server

    Peszynska, Malgorzata; Iltis, Gabriel; Schlueter, Steffen; Wildenschild, Dorthe

    2015-01-01

    Biofilm growth changes many physical properties of porous media such as porosity, permeability and mass transport parameters. The growth depends on various environmental conditions, and in particular, on flow rates. Modeling the evolution of such properties is difficult both at the porescale where the phase morphology can be distinguished, as well as during upscaling to the corescale effective properties. Experimental data on biofilm growth is also limited because its collection can interfere with the growth, while imaging itself presents challenges. In this paper we combine insight from imaging, experiments, and numerical simulations and visualization. The experimental dataset is based on glass beads domain inoculated by biomass which is subjected to various flow conditions promoting the growth of biomass and the appearance of a biofilm phase. The domain is imaged and the imaging data is used directly by a computational model for flow and transport. The results of the computational flow model are upscaled to...

  10. Sfp-type PPTase inactivation promotes bacterial biofilm formation and ability to enhance wheat drought tolerance

    Directory of Open Access Journals (Sweden)

    Salme eTimmusk

    2015-05-01

    Full Text Available Paenibacillus polymyxa is a common soil bacterium with broad range of practical applications. An important group of secondary metabolites in P. polymyxa are nonribosomal peptide and polyketide derived metabolites (NRP/PK. Modular nonribosomal peptide synthetases catalyse main steps in the biosynthesis of the complex secondary metabolites. Here we report on the inactivation of an A26 sfp-type phosphopantetheinyl transferase. The inactivation of the gene resulted in loss of NRP/PK production. In contrast to the former Bacillus spp. model the mutant strain compared to wild type showed greatly enhanced biofilm formation ability. Its biofilm promotion is directly mediated by NRP/PK, as exogenous addition of the wild type metabolite extracts restores its biofilm formation level. Wheat inoculation with bacteria that had lost their sfp-type PPTase gene resulted in two times higher plant survival and about three times increased biomass under severe drought stress compared to wild type.

  11. Linear surface roughness growth and flow smoothening in a three-dimensional biofilm model

    CERN Document Server

    Head, D A

    2013-01-01

    The sessile microbial communities known as biofilms exhibit different surface structures as environmental factors are varied, including nutrient availability and flow-generated shear stresses. Here we modify an established agent-based biofilm model to include adhesive interactions, permitting it to mechanically react to an imposed stress. This model is employed to analyse the growth of surface roughness of single-species, three-dimensional biofilms. We find linear growth laws of surface geometry in both horizontal and vertical directions, and an active surface layer whose thickness anti-correlates with roughness. Flow is consistently shown to reduce surface roughness without affecting the active layer. We argue that the rapid roughening is due to non-local surface interactions mediated by the nutrient field which are curtailed by sufficiently rapid flows, and suggest simplified models will need to be developed to elucidate the underlying mechanisms.

  12. Wireless transmission of fast-scan cyclic voltammetry at a carbon-fiber microelectrode: proof of principle.

    Science.gov (United States)

    Garris, Paul A; Ensman, Robert; Poehlman, John; Alexander, Andy; Langley, Paul E; Sandberg, Stefan G; Greco, Phillip G; Wightman, R Mark; Rebec, George V

    2004-12-30

    Fast-scan cyclic voltammetry (FSCV) at a carbon-fiber microelectrode (CFM) provides exquisite temporal and spatial resolution for monitoring brain chemistry. The utility of this approach has recently been demonstrated by measuring sub-second dopamine changes associated with behavior. However, one drawback is the cable link between animal and recording equipment that restricts behavior and precludes monitoring in complex environments. As a first step towards developing new instrumentation to overcome this technical limitation, the goal of the present study was to establish proof of principle for the wireless transmission of FSCV at a CFM. Proof of principle was evaluated in terms of measurement stability, fidelity, and susceptibility to ambient electrical noise. Bluetooth digital telemetry provided bi-directional communication between remote and home-base units and stable, high-fidelity data transfer comparable to conventional, wired systems when tested using a dummy cell (i.e., a resistor and capacitor in series simulating electrical properties of a CFM), and dopamine measurements with flow injection analysis and in the anesthetized rat with electrical stimulation. The wireless system was also less susceptible to interference from ambient electrical noise. Taken together, the present findings establish proof of principle for the wireless transmission of FSCV at a CFM.

  13. Unveiling One-Dimensional Supramolecular Structures Formed Through π-π Stacking of Phenothiazines by Differential Pulse Voltammetry.

    Science.gov (United States)

    Carvalho, Fernando R; Zampieri, Eduardo H; Caetano, Wilker; Silva, Rafael

    2017-03-09

    Organic based nanomaterials can be self-assembled by strong and directional intermolecular forces as π-π interactions are. Experimental information about the stability, size and geometry of those formed structures are very limited for species which easily aggregates even at very low concentration. Differential pulse voltammetry can unveil the formation, growth and also the stability window of ordered one-dimensional, lamellar, self-aggregates formed by supramolecular π stacking of phenothiazines at micromolar (10-6 mol·L-1) concentration. The self-diffusion features of the species at different concentration were determined by DPV and used to probe the π staking process through the concept of the frictional resistance. It is observed that toluidine blue and methylene blue start to self-aggregate around 9 µmol·L-1 and the self-aggregation process occurs by one-dimensional growth as the concentration of the phenothiazines is increased up to around 170 µmol·L-1 for toluidine blue O and 200 µmol·L-1 for methylene blue. At higher concentration the aggregation process leads to structures with lower anisometry.

  14. Substituent Inductive Effects on the Electrochemical Oxidation of Flavonoids Studied by Square Wave Voltammetry and Ab Initio Calculations.

    Science.gov (United States)

    Arroyo-Currás, Netzahualcóyotl; Rosas-García, Víctor M; Videa, Marcelo

    2016-10-27

    Flavonoids are natural products commonly found in the human diet that show antioxidant, anti-inflammatory and anti-hepatotoxic activities. These nutraceutical properties may relate to the electrochemical activity of flavonoids. To increase the understanding of structure-electrochemical activity relations and the inductive effects that OH substituents have on the redox potential of flavonoids, we carried out square-wave voltammetry experiments and ab initio calculations of eight flavonoids selected following a systematic variation in the number of hydroxyl substituents and their location on the flavan backbone: three flavonols, three anthocyanidins, one anthocyanin and the flavonoid backbone flavone. We compared the effect that the number of -OH groups in the ring B of flavan has on the oxidation potential of the flavonoids considered, finding linear correlations for both flavonols and anthocyanidins ( R 2 = 0.98 ). We analyzed the effects that position and number of -OH substituents have on electron density distributions via ab initio quantum chemical calculations. We present direct correlations between structural features and oxidation potentials that provide a deeper insight into the redox chemistry of these molecules.

  15. Probing the electrochemical properties of TiO{sub 2}/graphene composite by cyclic voltammetry and impedance spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Yadav, Pankaj [Department of Physics, Marwadi Education Foundation, Rajkot 360003 (India); Pandey, Kavita; Bhatt, Parth [School of Solar Energy, Pandit Deendayal Petroleum University, Gandhinagar 382007 (India); Tripathi, Brijesh, E-mail: brijesh.tripathi@sse.pdpu.ac.in [School of Technology, Pandit Deendayal Petroleum University, Gandhinagar 382007 (India); Pandey, Manoj Kumar; Kumar, Manoj [School of Technology, Pandit Deendayal Petroleum University, Gandhinagar 382007 (India)

    2016-04-15

    Highlights: • Role of TiO{sub 2}/graphene composite in charge transport within supercapacitors. • DC and AC characterization to investigate voltage dependence of charge transport. • Physical insight into the electrochemistry of electrode–electrolyte interface. - Abstract: This work describes the role of graphene in charge transport and diffusion mechanism at TiO{sub 2}/graphene electrode–electrolyte interface. To explore the mechanism, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used. The CV results depict that TiO{sub 2} and TiO{sub 2}/graphene electrodes behave differently in terms of charge transport and ion adsorption under the steady state conditions. The performance of TiO{sub 2} electrode–electrolyte interface is mainly limited by the charge transport and pseudo-capacitive effects while the response of TiO{sub 2}/graphene electrode–electrolyte interface is mainly dominated by the double layer capacitive effects. The EIS measurement leads to the direct determination of broad range of parameters, i.e. series resistance, charge transport, rate capability and ion diffusion. The experimental results and their analysis will have a significant impact on understanding the role of graphene in the electrochemical response of an electrode–electrolyte interface.

  16. Characterization of starvation-induced dispersion in Pseudomonas putida biofilms

    DEFF Research Database (Denmark)

    Gjermansen, Morten; Ragas, Paula Cornelia; Sternberg, Claus;

    2005-01-01

    that they must be able to regulate their ability to form biofilm and to dissolve biofilm. We present an investigation of a biofilm dissolution process occurring in flow-chamber-grown Pseudomonas putida biofilms. Local starvation-induced biofilm dissolution appears to be an integrated part of P. putida biofilm...... development that causes characteristic structural rearrangements. Rapid global dissolution of entire P. putida biofilms was shown to occur in response to carbon starvation. Genetic analysis suggested that the adjacent P. putida genes PP0164 and PP0165 play a role in P. putida biofilm formation and dissolution....... PP0164 encodes a putative periplasmic protein of previously unknown function, and PP0164 mutant bacteria are sticky, and unable to reduce their adhesiveness and dissolve their biofilm in response to carbon starvation. PP0165 encodes a putative transmembrane protein containing GGDEF and EAL domains...

  17. An improved protocol for harvesting Bacillus subtilis colony biofilms.

    Science.gov (United States)

    Fuchs, Felix Matthias; Driks, Adam; Setlow, Peter; Moeller, Ralf

    2017-03-01

    Bacterial biofilms cause severe problems in medicine and industry due to the high resistance to disinfectants and environmental stress of organisms within biofilms. Addressing challenges caused by biofilms requires full understanding of the underlying mechanisms for bacterial resistance and survival in biofilms. However, such work is hampered by a relative lack of systems for biofilm cultivation that are practical and reproducible. To address this problem, we developed a readily applicable method to culture Bacillus subtilis biofilms on a membrane filter. The method results in biofilms with highly reproducible characteristics, and which can be readily analyzed by a variety of methods with little further manipulation. This biofilm preparation method simplifies routine generation of B. subtilis biofilms for molecular and cellular analysis, and could be applicable to other microbial systems.

  18. Acetic acid bacteria from biofilm of strawberry vinegar visualized by microscopy and detected by complementing culture-dependent and culture-independent techniques.

    Science.gov (United States)

    Valera, Maria José; Torija, Maria Jesús; Mas, Albert; Mateo, Estibaliz

    2015-04-01

    Acetic acid bacteria (AAB) usually develop biofilm on the air-liquid interface of the vinegar elaborated by traditional method. This is the first study in which the AAB microbiota present in a biofilm of vinegar obtained by traditional method was detected by pyrosequencing. Direct genomic DNA extraction from biofilm was set up to obtain suitable quality of DNA to apply in culture-independent molecular techniques. The set of primers and TaqMan--MGB probe designed in this study to enumerate the total AAB population by Real Time--PCR detected between 8 × 10(5) and 1.2 × 10(6) cells/g in the biofilm. Pyrosequencing approach reached up to 10 AAB genera identification. The combination of culture-dependent and culture-independent molecular techniques provided a broader view of AAB microbiota from the strawberry biofilm, which was dominated by Ameyamaea, Gluconacetobacter, and Komagataeibacter genera. Culture-dependent techniques allowed isolating only one genotype, which was assigned into the Ameyamaea genus and which required more analysis for a correct species identification. Furthermore, biofilm visualization by laser confocal microscope and scanning electronic microscope showed different dispositions and cell morphologies in the strawberry vinegar biofilm compared with a grape vinegar biofilm.

  19. Phototrophic microorganisms in biofilm samples from Vernjika Cave, Serbia

    Science.gov (United States)

    Popović, Slađana; Jovanović, Jelena; Predojević, Dragana; Trbojević, Ivana; Blagojević, Ana; Subakov Simić, Gordana

    2016-04-01

    Caves represent specific natural monuments in terms of structure, complexity and beauty which can be found worldwide. Even though they are considered extreme environments, they are still a unique habitat for a large number of organisms that grow and proliferate here. Often can be seen that the cave walls are differently coloured as a consequence of the biofilm development. Biofilms represent complex communities of microorganisms that can develop on different kind of surfaces, including various rock surfaces. Each microbe species play a different role in a community, but their development on stone surfaces can cause substantial damage to the substrates through different mechanisms of biodeterioration and degradation. There is an increased interest in the phototrophic component of biofilms (aerophytic cyanobacteria and algae), especially cyanobacteria, an ancient microorganisms capable to survive the most diverse extreme conditions. These phototrophs can easily be found at cave entrances illuminated by direct or indirect sunlight and areas near artificial lights. Cyanobacteria and algae were investigated in biofilm samples taken from the entrance of Vernjika Cave in Eastern Serbia. Cyanobacteria, Chlorophyta and Bacillariophyta were documented, with Cyanobacteria as a group with the highest number of recorded taxa. Chroococcalean species were the most diverse with the most frequently encountered species from the genus Gloeocapsa. Phormidium and Nostoc species were commonly recorded Oscillatoriales and Nostocles, respectively. Among Oscillatoriales species, it was noticed that one Phormidium species precipitates CaCO3 on it's sheats. Trebouxia sp. and Desmococcus olivaceus were frequently documented Chlorophyta, and representatives of Bacillariophyta were exclusively aerophytic taxa, mostly belonging to the genera Luticola and Humidophila. Measured ecological parameters, temperature and relative humidity, were influenced by the external climatic changes, while light

  20. Susceptibility of Porphyromonas gingivalis in biofilms to amoxicillin, doxycycline and metronidazole

    DEFF Research Database (Denmark)

    Larsen, T.

    2002-01-01

    Biofilm, Porphyromonas gingivalis, susceptibility testing, amoxicillin, doxycycline, metronidazole......Biofilm, Porphyromonas gingivalis, susceptibility testing, amoxicillin, doxycycline, metronidazole...