Sample records for h2-based membrane biofilm

  1. The membrane biofilm reactor: the natural partnership of membranes and biofilm.

    PubMed

    Rittmann, B E

    2006-01-01

    Many exciting new technologies for water-quality control combine microbiological processes with adsorption, advanced oxidation, a membrane or an electrode to improve performance, address emerging contaminants or capture renewable energy. An excellent example is the H2-based membrane biofilm reactor (MBfR), which delivers H2 gas to a biofilm that naturally accumulates on the outer surface of a bubbleless membrane. Autotrophic bacteria in the biofilm oxidise the H2 and use the electrons to reduce NO3-, CIO4- and other oxidised contaminants. This natural partnership of membranes and biofilm makes it possible to gain many cost, performance and simplicity advantages from using H2 as the electron donor for microbially catalysed reductions. The MBfR has been demonstrated for denitrification in drinking water; reduction of perchlorate in groundwater; reduction of selenate, chromate, trichloroethene and other emerging contaminants; advanced N removal in wastewater treatment and autotrophic total-N removal.

  2. Combination of cupric ion with hydroxylamine and hydrogen peroxide for the control of bacterial biofilms on RO membranes.

    PubMed

    Lee, Hye-Jin; Kim, Hyung-Eun; Lee, Changha

    2017-03-01

    Combinations of Cu(II) with hydroxylamine (HA) and hydrogen peroxide (H 2 O 2 ) (i.e., Cu(II)/HA, Cu(II)/H 2 O 2 , and Cu(II)/HA/H 2 O 2 systems) were investigated for the control of P. aeruginosa biofilms on reverse osmosis (RO) membranes. These Cu(II)-based disinfection systems effectively inactivated P. aeruginosa cells, exhibiting different behaviors depending on the state of bacterial cells (planktonic or biofilm) and the condition of biofilm growth and treatment (normal or pressurized condition). The Cu(II)/HA and Cu(II)/HA/H 2 O 2 systems were the most effective reagents for the inactivation of planktonic cells. However, these systems were not effective in inactivating cells in biofilms on the RO membranes possibly due to the interactions of Cu(I) with extracellular polymeric substances (EPS), where biofilms were grown and treated in center for disease control (CDC) reactors. Different from the results using CDC reactors, in a pressurized cross-flow RO filtration unit, the Cu(II)/HA/H 2 O 2 treatment significantly inactivated biofilm cells formed on the RO membranes, successfully recovering the permeate flux reduced by the biofouling. The pretreatment of feed solutions by Cu(II)/HA and Cu(II)/HA/H 2 O 2 systems (applied before the biofilm formation) effectively mitigated the permeate flux decline by preventing the biofilm growth on the RO membranes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Assessing microbial competition in a hydrogen-based membrane biofilm reactor (MBfR) using multidimensional modeling.

    PubMed

    Martin, Kelly J; Picioreanu, Cristian; Nerenberg, Robert

    2015-09-01

    The membrane biofilm reactor (MBfR) is a novel technology that safely delivers hydrogen to the base of a denitrifying biofilm via gas-supplying membranes. While hydrogen is an effective electron donor for denitrifying bacteria (DNB), it also supports sulfate-reducing bacteria (SRB) and methanogens (MET), which consume hydrogen and create undesirable by-products. SRB and MET are only competitive for hydrogen when local nitrate concentrations are low, therefore SRB and MET primarily grow near the base of the biofilm. In an MBfR, hydrogen concentrations are greatest at the base of the biofilm, making SRB and MET more likely to proliferate in an MBfR system than a conventional biofilm reactor. Modeling results showed that because of this, control of the hydrogen concentration via the intramembrane pressure was a key tool for limiting SRB and MET development. Another means is biofilm management, which supported both sloughing and erosive detachment. For the conditions simulated, maintaining thinner biofilms promoted higher denitrification fluxes and limited the presence of SRB and MET. The 2-d modeling showed that periodic biofilm sloughing helped control slow-growing SRB and MET. Moreover, the rough (non-flat) membrane assembly in the 2-d model provided a special niche for SRB and MET that was not represented in the 1-d model. This study compared 1-d and 2-d biofilm model applicability for simulating competition in counter-diffusional biofilms. Although more computationally expensive, the 2-d model captured important mechanisms unseen in the 1-d model. © 2015 Wiley Periodicals, Inc.

  4. Enhancing biodegradation of C16-alkyl quaternary ammonium compounds using an oxygen-based membrane biofilm reactor.

    PubMed

    Lai, YenJung Sean; Ontiveros-Valencia, Aura; Ilhan, Zehra Esra; Zhou, Yun; Miranda, Evelyn; Maldonado, Juan; Krajmalnik-Brown, Rosa; Rittmann, Bruce E

    2017-10-15

    Quaternary ammonium compounds (QACs) (e.g., hexadecyltrimethyl-ammonium bromide, CTAB) are emerging contaminants with widespread use as surfactants and disinfectants. Because the initial step of QAC biodegradation is mono-oxygenation, QAC degraders require O 2 , but normal aeration leads to serious foaming. Here, we developed and tested an oxygen-based membrane biofilm reactor (O 2 -MBfR) that delivers O 2 by diffusion through the walls of hollow-membranes to a biofilm accumulating on the outer surface of membranes. The O 2 -MBfR sustained QAC biodegradation even with high and toxic QAC input concentrations, up to 400 mg/L CTAB. Bubbleless O 2 transfer completely eliminated foaming, and biofilm accumulation helped the QAC biodegraders resist toxicity. Pseudomonas, Achromobacter, Stenotrophomonas, and members of the Xanthomonadaceae family were dominant in the biofilm communities degrading CTAB, and their proportions depended on the O 2 -delivery capacity of the membranes. Bacteria capable of biodegrading QACs often harbor antibiotic resistance genes (ARGs) that help them avoid QAC toxicity. Gene copies of ARGs were detected in biofilms and liquid, but the levels of ARGs were 5- to 35-fold lower in the liquid than in the biofilm. In summary, the O 2 -MBfR achieved aerobic biodegradation of CTAB with neither foaming nor toxicity, and it also minimized the spread of ARGs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Bio-reduction of N-nitrosodimethylamine (NDMA) using a hydrogen-based membrane biofilm reactor.

    PubMed

    Chung, Jinwook; Ahn, Chang-Hoon; Chen, Zhuo; Rittmann, Bruce E

    2008-01-01

    N-Nitrosodimethylamine (NDMA) is a disinfection by-product shown to be carcinogenic, mutagenic, and teratogenic. A feasible detoxification pathway for NDMA is a three-step bio-reduction that leads to ammonia and dimethylamine. This study examines the bio-reduction of NDMA in a H2-based membrane biofilm reactor (MBfR) that also is active in nitrate and sulfate reductions. In particular, the study investigates the effects of H2 availability and the relative loadings of NDMA, nitrate, and sulfate, which potentially are competing electron acceptors. The results demonstrate that NDMA was bio-reduced to a major extent (i.e., at least 96%) in a H2-based MBfR in which the electron-equivalent fluxes from H2 oxidation were dominated by nitrate and sulfate reductions. NDMA reduction kinetics responded to NDMA concentration, H2 pressure, and the presence of competing acceptors. The most important factor controlling NDMA-reduction kinetics was the H2 availability, controlled primarily by the H2 pressure, and secondarily by competition from nitrate reduction.

  6. Bioreduction of para-chloronitrobenzene in a hydrogen-based hollow-fiber membrane biofilm reactor: effects of nitrate and sulfate.

    PubMed

    Li, Haixiang; Zhang, Zhiqiang; Xu, Xiaoyin; Liang, Jun; Xia, Siqing

    2014-04-01

    A continuous-stirred, hydrogen-based, hollow-fiber membrane biofilm reactor (HFMBfR) that was active in nitrate and sulfate reductions was shown to be effective for degradation or detoxification of para-chloronitrobenzene (p-CNB) in water by biotransforming it first to para-chloroaniline (nitro-reduction) and then to aniline (reductive dechlorination) with hydrogen (H2) as an electron donor. A series of short-term experiments examined the effects of nitrate and sulfate on p-CNB bioreduction. The results obtained showed both higher nitrate and sulfate concentration declined the p-CNB bioreduction in the biofilm, and this suggests the competition for H2 caused less H2 available for the p-CNB bioreduction when the H2 demand for the reductions was larger. Denitrification and sulfate reduction intermediates were thought to be potential factors inhibiting the p-CNB bioreduction. Analysis of electron-equivalent fluxes and reaction orders in the biofilm further demonstrated both denitrification and sulfate reduction competed more strongly for H2 availability than p-CNB bioreduction. These findings have significant implications for the HFMBfR used for degrading p-CNB under denitrifying and/or sulfate reducing conditions.

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

    PubMed

    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(-2h(-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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Utilizing a one-dimensional multispecies model to simulate the nutrient reduction and biomass structure in two types of H2-based membrane-aeration biofilm reactors (H2-MBfR): model development and parametric analysis.

    PubMed

    Wang, Zuowei; Xia, Siqing; Xu, Xiaoyin; Wang, Chenhui

    2016-02-01

    In this study, a one-dimensional multispecies model (ODMSM) was utilized to simulate NO3(-)-N and ClO4(-) reduction performances in two kinds of H2-based membrane-aeration biofilm reactors (H2-MBfR) within different operating conditions (e.g., NO3(-)-N/ClO4(-) loading rates, H2 partial pressure, etc.). Before the simulation process, we conducted the sensitivity analysis of some key parameters which would fluctuate in different environmental conditions, then we used the experimental data to calibrate the more sensitive parameters μ1 and μ2 (maximum specific growth rates of denitrification bacteria and perchlorate reduction bacteria) in two H2-MBfRs, and the diversity of the two key parameters' values in two types of reactors may be resulted from the different carbon source fed in the reactors. From the simulation results of six different operating conditions (four in H2-MBfR 1 and two in H2-MBfR 2), the applicability of the model was approved, and the variation of the removal tendency in different operating conditions could be well simulated. Besides, the rationality of operating parameters (H2 partial pressure, etc.) could be judged especially in condition of high nutrients' loading rates. To a certain degree, the model could provide theoretical guidance to determine the operating parameters on some specific conditions in practical application.

  9. Membrane-aerated biofilm proton and oxygen flux during chemical toxin exposure.

    PubMed

    McLamore, E S; Zhang, W; Porterfield, D M; Banks, M K

    2010-09-15

    Bioreactors containing sessile bacteria (biofilms) grown on hollow fiber membranes have been used for treatment of many wastestreams. Real time operational control of bioreactor performance requires detailed knowledge of the relationship between bulk liquid water quality and physiological transport at the biofilm-liquid interface. Although large data sets exist describing membrane-aerated bioreactor effluent quality, very little real time data is available characterizing boundary layer transport under physiological conditions. A noninvasive, microsensor technique was used to quantify real time (≈1.5 s) changes in oxygen and proton flux for mature Nitrosomonas europaea and Pseudomonas aeruginosa biofilms in membrane-aerated bioreactors following exposure to environmental toxins. Stress response was characterized during exposure to toxins with known mode of action (chlorocarbonyl cyanide phenyl-hydrazone and potassium cyanide), and four environmental toxins (rotenone, 2,4-dinitrophenol, cadmium chloride, and pentachlorophenol). Exposure to sublethal concentrations of all environmental toxins caused significant increases in O(2) and/or H(+) flux (depending on the mode of action). These real time microscale signatures (i.e., fingerprints) of O(2) and H(+) flux can be coupled with bulk liquid analysis to improve our understanding of physiology in counter-diffusion biofilms found within membrane aerated bioreactors; leading to enhanced monitoring/modeling strategies for bioreactor control.

  10. Bacterial community structure and activity of sulfate reducing bacteria in a membrane aerated biofilm analyzed by microsensor and molecular techniques.

    PubMed

    Liu, Hong; Tan, Shuying; Sheng, Zhiya; Liu, Yang; Yu, Tong

    2014-11-01

    The activities and vertical spatial distribution of sulfate reducing bacteria (SRB) in an oxygen (O2 )-based membrane aerated biofilm (MAB) were investigated using microsensor (O2 and H2 S) measurements and molecular techniques (polymerase chain reaction-denaturing gradient gel electrophoresis [PCR-DGGE] and fluorescence in situ hybridization [FISH]). The O2 concentration profile revealed that O2 penetrated from the bottom (substratum) of the gas permeable membrane, and was gradually consumed within the biofilm until it was completely depleted near the biofilm/bulk liquid interface, indicating oxic and anoxic zone in the MAB. The H2 S concentration profile showed that H2 S production was found in the upper 285 µm of the biofilm, indicating a high activity of SRB in this region. The results from DGGE of the PCR-amplified dissimilatory sulfite reductase subunit B (dsrB) gene and FISH showed an uneven spatial distribution of SRB. The maximum SRB biomass was located in the upper biofilm. The information from the molecular analysis can be supplemented with that from microsensor measurements to better understand the microbial community and activity of SRB in the MAB. © 2014 Wiley Periodicals, Inc.

  11. Simultaneous removal of selected oxidized contaminants in groundwater using a continuously stirred hydrogen-based membrane biofilm reactor.

    PubMed

    Xia, Siqing; Liang, Jun; Xu, Xiaoyin; Shen, Shuang

    2013-01-01

    A laboratory trial was conducted for evaluating the capability of a continuously stirred hydrogen-based membrane biofilm reactor to simultaneously reduce nitrate (NO(3-)-N), sulfate (SO4(2-)), bromate (BrO3-), hexavalent chromium (Cr(VI)) and parachloronitrobenzene (p-CNB). The reactor contained two bundles of hollow fiber membranes functioning as an autotrophic biofilm carrier and hydrogen pipe as well. On the condition that hydrogen was supplied as electron donor and diffused into water through membrane pores, autohydrogenotrophic bacteria were capable of reducing contaminants to forms with lower toxicity. Reduction occurred within 1 day and removal fluxes for NO(3-)-N, SO4(2-), BrO3-, Cr(VI), and p-CNB reached 0.641, 2.396, 0.008, 0.016 and 0.031 g/(day x m2), respectively after 112 days of continuous operation. Except for the fact that sulfate was 37% removed under high surface loading, the other four contaminants were reduced by over 95%. The removal flux comparison between phases varying in surface loading and H2 pressure showed that decreasing surface loading or increasing H2 pressure would promote removal flux. Competition for electrons occurred among the five contaminants. Electron-equivalent flux analysis showed that the amount of utilized hydrogen was mainly controlled by NO(3-)-N and SO4(2-) reduction, which accounted for over 99% of the electron flux altogether. It also indicated the electron acceptor order, showing that nitrate was the most prior electron acceptor while suIfate was the second of the five contaminants.

  12. A confocal microscopy based method to monitor extracellular pH in fungal biofilms.

    PubMed

    Schlafer, Sebastian; Kamp, Anja; Garcia, Javier E

    2018-04-19

    pH in fungal biofilms is important for a variety of fungal infections and industrial applications involving fungal biofilms, but to date, it has never been measured directly inside the biofilm matrix. In the present study, a new methodology was developed allowing for confocal microscopy based monitoring of extracellular pH inside fungal biofilms. Monospecies biofilms of Aspergillus fumigatus, Candida albicans, Candida dubliniensis and Cryptococcus neoformans were stained with the pH dependent ratiometric probe C-SNARF-4, imaged with a confocal microscope, and a digital image analysis procedure was developed to determine pH in the extracellular matrix. As a proof of concept, pH developments at the biofilm-substratum interface were monitored for one h after exposure to glucose. Observed pH drops differed considerably between the different species and also between replicate biofilms of the same species. C. albicans biofilms showed the highest acidogenicity, with pH drops occurring much faster than in planktonic culture. pH ratiometry with C-SNARF-4 is a valuable tool to get insight into fungal biofilm metabolism and may shed new light on both disease-related and industrially relevant processes in fungal biofilms.

  13. Membrane-aerated biofilm reactor for the removal of 1,2-dichloroethane by Pseudomonas sp. strain DCA1.

    PubMed

    Hage, J C; Van Houten, R T; Tramper, J; Hartmans, S

    2004-06-01

    A membrane-aerated biofilm reactor (MBR) with a biofilm of Pseudomonas sp. strain DCA1 was studied for the removal of 1,2-dichloroethane (DCA) from water. A hydrophobic membrane was used to create a barrier between the liquid and the gas phase. Inoculation of the MBR with cells of strain DCA1 grown in a continuous culture resulted in the formation of a stable and active DCA-degrading biofilm on the membrane. The maximum removal rate of the MBR was reached at a DCA concentration of approximately 80 micro M. Simulation of the DCA fluxes into the biofilm showed that the MBR performance at lower concentrations was limited by the DCA diffusion rate rather than by kinetic constraints of strain DCA1. Aerobic biodegradation of DCA present in anoxic water could be achieved by supplying oxygen solely from the gas phase to the biofilm grown on the liquid side of the membrane. As a result, direct aeration of the water, which leads to undesired coagulation of iron oxides, could be avoided.

  14. Anaerobic granule-based biofilms formation reduces propionate accumulation under high H2 partial pressure using conductive carbon felt particles.

    PubMed

    Xu, Heng; Wang, Cuiping; Yan, Kun; Wu, Jing; Zuo, Jiane; Wang, Kaijun

    2016-09-01

    Syngas based co-digestion is not only more economically attractive than separate syngas methanation but also able to upgrade biogas and increase overall CH4 amount simultaneously. However, high H2 concentration in the syngas could inhibit syntrophic degradation of propionate, resulting in propionate accumulation and even failure of the co-digestion system. In an attempt to reduce propionate accumulation via enhancing both H2 interspecies transfer (HIT) and direct interspecies electron transfer (DIET) pathways, layered granule-based biofilms induced by conductive carbon felt particles (CCFP) was employed. The results showed that propionate accumulation was effectively reduced with influent COD load up to 7gL(-1)d(-1). Two types of granule-based biofilms, namely biofilm adhered to CCFP (B-CCFP) and granules formed by self-immobilization (B-SI) were formed in the reactor. Clostridium, Syntrophobacter, Methanospirillum were possibly involved in HIT and Clostridium, Geobacter, Anaerolineaceae, Methanosaeta in DIET, both of which might be responsible for the high-rate propionate degradation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Biological control of biofilms on membranes by metazoans.

    PubMed

    Klein, Theresa; Zihlmann, David; Derlon, Nicolas; Isaacson, Carl; Szivak, Ilona; Weissbrodt, David G; Pronk, Wouter

    2016-01-01

    Traditionally, chemical and physical methods have been used to control biofouling on membranes by inactivating and removing the biofouling layer. Alternatively, the permeability can be increased using biological methods while accepting the presence of the biofouling layer. We have investigated two different types of metazoans for this purpose, the oligochaete Aelosoma hemprichi and the nematode Plectus aquatilis. The addition of these grazing metazoans in biofilm-controlled membrane systems resulted in a flux increase of 50% in presence of the oligochaetes (Aelosoma hemprichi), and a flux increase of 119-164% in presence of the nematodes (Plectus aquatilis) in comparison to the control system operated without metazoans. The change in flux resulted from (1) a change in the biofilm structure, from a homogeneous, cake-like biofilm to a more heterogeneous, porous structure and (2) a significant reduction in the thickness of the basal layer. Pyrosequencing data showed that due to the addition of the predators, also the community composition of the biofilm in terms of protists and bacteria was strongly affected. The results have implications for a range of membrane processes, including ultrafiltration for potable water production, membrane bioreactors and reverse osmosis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Removal of gaseous trichloroethylene (TCE) in a composite membrane biofilm reactor.

    PubMed

    Kumar, Amit; Vercruyssen, Aline; Dewulf, Jo; Lens, Piet; Van Langenhove, Herman

    2012-01-01

    A membrane biofilm reactor (MBfR) was investigated for the degradation of trichloroethylene (TCE) vapors inoculated by Burkholderia vietnamiensis G4. Toluene (TOL) was used as the primary substrate. The MBfR was loaded sequentially with TOL, TCE (or both) during 110 days. In this study, a maximum steady-state TCE removal efficiency of 23% and a maximum volumetric elimination capacity (EC) of 2.1 g m(-3) h(-1) was achieved. A surface area based maximum elimination capacity (EC(m)) of 4.2 × 10(-3) g m(-2) h(-1) was observed, which is 2-10 times higher than reported in other gas phase biological treatment studies. However, further research is needed to optimize the TCE feeding cycle and to evaluate the inhibiting effects of TCE and its intermediates on TOL biodegradation.

  17. An extractive membrane biofilm reactor as alternative technology for the treatment of methyl tert-butyl ether contaminated water.

    PubMed

    Guisado, I M; Purswani, J; González-López, J; Pozo, C

    2016-09-01

    Among the strategies developed for contaminated groundwater bioremediation, those based on the use of bacteria adhering to inert supports and establishing biofilms have gained great importance in this field. Extractive membrane biofilm reactor (EMBFR) technology offers productive solutions for the removal of volatile and semi-volatile compounds. EMBFR technology is based on the use of extractive semipermeable membranes through which contaminants migrate to the biological compartment in which microorganisms with pollutant biotransformation and/or mineralization capacities can grow, forming an active biofilm on the membrane surface. The objective of this study was to assess the use of three bacterial strains (Paenibacillus sp. SH7 CECT 8558, Agrobacterium sp. MS2 CECT 8557, and Rhodococcus ruber EE6 CECT 8612), as inoculum in a lab-scale EMBFR running for 28 days under aerobic conditions to eliminate methyl tert-butyl ether (MTBE) from water samples. Three different hydraulic retention times (1, 6, and 12 h) were employed. MTBE degradation values were determined daily by a gas GC-MS technique, as well as suspended bacterial growth. The biofilm established by the bacterial strains on the semipermeable membrane was detected by Field-Emission Scanning Electron Microscopy (FESEM) at the end of each experiment. The acute toxicity of the treated effluents and biomedium was determined by Microtox © assay (EC 50 ).The results achieved from the MTBE degradation, biofilm formation, and toxicity analysis indicated that bacterial strains MS2 and EE6 were the best options as selective inoculum, although further research is needed, particularly with regard to their possible use as a mixed culture. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1238-1245, 2016. © 2016 American Institute of Chemical Engineers.

  18. Impact of oxygen on the coexistence of nitrification, denitrification, and sulfate reduction in oxygen-based membrane aerated biofilm.

    PubMed

    Liu, Hong; Tan, Shuying; Sheng, Zhiya; Yu, Tong; Liu, Yang

    2015-03-01

    Membrane aerated biofilms (MABs) are subject to "counter diffusion" of oxygen and substrates. In a membrane aerated biofilm reactor, gases (e.g., oxygen) diffuse through the membrane into the MAB, and liquid substrates pass from the bulk liquid into the MAB. This behavior can result in a unique biofilm structure in terms of microbial composition, distribution, and community activity in the MAB. Previous studies have shown simultaneous aerobic oxidation, nitrification, and denitrification within a single MAB. Using molecular techniques, we investigated the growth of sulfate-reducing bacteria (SRB) in the oxygen-based MAB attached to a flat sheet membrane. Denaturing gradient gel electrophoresis of the amplified 16S rRNA gene fragments and functional gene fragments specific for ammonia-oxidizing bacteria (amoA), denitrifying bacteria (nirK), and SRB (dsrB) demonstrated the coexistence of nitrifiers, denitrifiers, and SRB communities within a single MAB. The functional diversities of SRB and denitrifiers decreased with an increase in the oxygen concentration in the bulk water of the reactor.

  19. Kinetic development of biofilm on NF membranes at the Méry-sur-Oise plant, France.

    PubMed

    Houari, Ahmed; Seyer, Damien; Kecili, Karima; Heim, Véronique; Martino, Patrick Di

    2013-01-01

    The kinetic formation of biofilms developing on nanofiltration (NF) membranes was studied for 2 years in the water production unit of Méry-sur-Oise, France. New membranes were set up in a pilot train integrated to the plant and autopsied after operation for 7, 80, 475 and 717 days. The biofouling layer was studied by confocal laser scanning microscope after 4',6-diamidino-2-phenyindole dihydrochloride and lectin staining, and by attenuated total reflectance-Fourier transform infrared spectroscopy and rheology experiments. Three stages of biofilm growth were discriminated: (1) the presence of sessile microcolonies embedded in an exopolymeric matrix (after filtration for seven days); (2) membrane coverage expansion through microcolony development and biofilm growth in three dimensions (up to 80 days filtration); and (3) biofilm maturation by densification (after filtration for 80-717 days). Biofilm maturation resulted in total coverage of the membrane surface and matrix residue diversification, development of the polysaccharide network, and the strengthening of matrix cohesion through viscosity and elasticity increases. The wettability and permeability of the fouled NF membranes decreased quickly and continuously throughout the biofilm development process. The longitudinal pressure drop (LPD) increased only after the biofilm reached a quantitative threshold. The decline in membrane permeability may be the result of contributions from many fouling mechanisms but the LPD was more substantially influenced by biofilm development.

  20. Proteome Profiles of Outer Membrane Vesicles and Extracellular Matrix of Pseudomonas aeruginosa Biofilms.

    PubMed

    Couto, Narciso; Schooling, Sarah R; Dutcher, John R; Barber, Jill

    2015-10-02

    In the present work, two different proteomic platforms, gel-based and gel-free, were used to map the matrix and outer membrane vesicle exoproteomes of Pseudomonas aeruginosa PAO1 biofilms. These two proteomic strategies allowed us a confident identification of 207 and 327 proteins from enriched outer membrane vesicles and whole matrix isolated from biofilms. Because of the physicochemical characteristics of these subproteomes, the two strategies showed complementarity, and thus, the most comprehensive analysis of P. aeruginosa exoproteome to date was achieved. Under our conditions, outer membrane vesicles contribute approximately 20% of the whole matrix proteome, demonstrating that membrane vesicles are an important component of the matrix. The proteomic profiles were analyzed in terms of their biological context, namely, a biofilm. Accordingly relevant metabolic processes involved in cellular adaptation to the biofilm lifestyle as well as those related to P. aeruginosa virulence capabilities were a key feature of the analyses. The diversity of the matrix proteome corroborates the idea of high heterogeneity within the biofilm; cells can display different levels of metabolism and can adapt to local microenvironments making this proteomic analysis challenging. In addition to analyzing our own primary data, we extend the analysis to published data by other groups in order to deepen our understanding of the complexity inherent within biofilm populations.

  1. The correlation between biofilm biopolymer composition and membrane fouling in submerged membrane bioreactors.

    PubMed

    Luo, Jinxue; Zhang, Jinsong; Tan, Xiaohui; McDougald, Diane; Zhuang, Guoqiang; Fane, Anthony G; Kjelleberg, Staffan; Cohen, Yehuda; Rice, Scott A

    2014-10-01

    Biofouling, the combined effect of microorganism and biopolymer accumulation, significantly reduces the process efficiency of membrane bioreactors (MBRs). Here, four biofilm components, alpha-polysaccharides, beta-polysaccharides, proteins and microorganisms, were quantified in MBRs. The biomass of each component was positively correlated with the transmembrane pressure increase in MBRs. Proteins were the most abundant biopolymer in biofilms and showed the fastest rate of increase. The spatial distribution and co-localization analysis of the biofouling components indicated at least 60% of the extracellular polysaccharide (EPS) components were associated with the microbial cells when the transmembrane pressure (TMP) entered the jump phase, suggesting that the EPS components were either secreted by the biofilm cells or that the deposition of these components facilitated biofilm formation. It is suggested that biofilm formation and the accumulation of EPS are intrinsically coupled, resulting in biofouling and loss of system performance. Therefore, strategies that control biofilm formation on membranes may result in a significant improvement of MBR performance.

  2. Succession of biofilm communities responsible for biofouling of membrane bio-reactors (MBRs)

    PubMed Central

    Luo, Jinxue; Lv, Pengyi; Zhang, Jinsong; Fane, Anthony G.; McDougald, Diane

    2017-01-01

    Biofilm formation is one of the main factors associated with membrane biofouling in membrane bioreactors (MBRs). As such, it is important to identify the responsible organisms to develop targeted strategies to control biofouling. This study investigated the composition and changes in the microbial communities fouling MBR membranes over time and correlated those changes with an increase in transmembrane pressure (TMP). Based on qPCR data, bacteria were the dominant taxa of the biofilm (92.9–98.4%) relative to fungi (1.5–6.9%) and archaea (0.03–0.07%). NMDS analysis indicated that during the initial stages of operation, the biofilm communities were indistinguishable from those found in the sludge. However, the biofilm community significantly diverged from the sludge over time and ultimately showed a unique biofilm profile. This suggested that there was strong selection for a group of organisms that were biofilm specialists. This pattern of succession and selection was correlated with the rapid increase in TMP, where bacteria including Rhodospirillales, Sphingomonadales and Rhizobiales dominated the biofilm at this time. While most of the identified fungal OTUs matched Candida sp., the majority of fungal communities were unclassified by 18S rRNA gene sequencing. Collectively, the data suggests that bacteria, primarily, along with fungi may play an important role in the rapid TMP increase and loss of system performance. PMID:28686622

  3. Calcium-Phosphate-Osteopontin Particles Reduce Biofilm Formation and pH Drops in in situ Grown Dental Biofilms.

    PubMed

    Schlafer, Sebastian; Ibsen, Casper J S; Birkedal, Henrik; Nyvad, Bente

    2017-01-01

    This 2-period crossover study investigated the effect of calcium-phosphate-osteopontin particles on biofilm formation and pH in 48-h biofilms grown in situ. Bovine milk osteopontin is a highly phosphorylated glycoprotein that has been shown to interfere with bacterial adhesion to salivary-coated surfaces. Calcium-phosphate-osteopontin particles have been shown to reduce biofilm formation and pH drops in a 5-species laboratory model of dental biofilm without affecting bacterial viability. Here, smooth surface biofilms from 10 individuals were treated ex vivo 6 times/day for 30 min with either calcium-phosphate-osteopontin particles or sterile saline. After growth, the amount of biofilm formed was determined by confocal microscopy, and pH drops upon exposure to glucose were monitored using confocal-microscopy-based pH ratiometry. A total of 160 biofilms were analysed. No adverse effects of repeated ex vivo treatment with calcium-phosphate-osteopontin particles were observed. Particle treatment resulted in a 32% lower amount of biofilm formed (p < 0.05), but large inter-individual differences could be observed. Biofilm pH was significantly higher upon particle treatment, both shortly after the addition of glucose and after 30 min of incubation with glucose (p < 0.05). Calcium-phosphate-osteopontin particles may represent a new therapeutic approach to caries control and aim at directly targeting virulence factors involved in the caries process. Further studies are required to determine the effect of particle treatment on more acidogenic/aciduric biofilms as well as the remineralizing potential of the particles. © 2016 S. Karger AG, Basel.

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

    PubMed

    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. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  5. In vitro antimicrobial effects and mechanism of atmospheric-pressure He/O2 plasma jet on Staphylococcus aureus biofilm

    NASA Astrophysics Data System (ADS)

    Xu, Zimu; Shen, Jie; Cheng, Cheng; Hu, Shuheng; Lan, Yan; Chu, Paul K.

    2017-03-01

    The antimicrobial effects and associated mechanism of inactivation of Staphylococcus aureus (S. aureus) NCTC-8325 biofilms induced by a He/O2 atmospheric-pressure plasma jet (APPJ) are investigated in vitro. According to CFU (colony forming units) counting and the resazurin-based assay, the 10 min He/O2 (0.5%) APPJ treatment produces the optimal inactivation efficacy (>5 log10 ml-1) against the S. aureus biofilm and 5% of the bacteria enter a viable but non-culturable (VBNC) state. Meanwhile, 94% of the bacteria suffer from membrane damage according to SYTO 9/PI counterstaining. Scanning electron microscopy (SEM) reveals that plasma exposure erodes the extracellular polymeric substances (EPS) and then the cellular structure. The H2DCFDA-stained biofilms show larger concentrations of intracellular reactive oxygen species (ROS) in membrane-intact bacteria with increasing plasma dose. The admixture of oxygen in the working gas highly contributes to the deactivation efficacy of the APPJ against S. aureus and the plasma-induced endogenous ROS may work together with the discharge-generated ROS to continuously damage the bacterial membrane structure leading to deactivation of the biofilm microbes.

  6. Architecture, component, and microbiome of biofilm involved in the fouling of membrane bioreactors.

    PubMed

    Inaba, Tomohiro; Hori, Tomoyuki; Aizawa, Hidenobu; Ogata, Atsushi; Habe, Hiroshi

    2017-01-01

    Biofilm formation on the filtration membrane and the subsequent clogging of membrane pores (called biofouling) is one of the most persistent problems in membrane bioreactors for wastewater treatment and reclamation. Here, we investigated the structure and microbiome of fouling-related biofilms in the membrane bioreactor using non-destructive confocal reflection microscopy and high-throughput Illumina sequencing of 16S rRNA genes. Direct confocal reflection microscopy indicated that the thin biofilms were formed and maintained regardless of the increasing transmembrane pressure, which is a common indicator of membrane fouling, at low organic-loading rates. Their solid components were primarily extracellular polysaccharides and microbial cells. In contrast, high organic-loading rates resulted in a rapid increase in the transmembrane pressure and the development of the thick biofilms mainly composed of extracellular lipids. High-throughput sequencing revealed that the biofilm microbiomes, including major and minor microorganisms, substantially changed in response to the organic-loading rates and biofilm development. These results demonstrated for the first time that the architectures, chemical components, and microbiomes of the biofilms on fouled membranes were tightly associated with one another and differed considerably depending on the organic-loading conditions in the membrane bioreactor, emphasizing the significance of alternative indicators other than the transmembrane pressure for membrane biofouling.

  7. The effect of PDT on H. influenzae biofilm in vitro

    NASA Astrophysics Data System (ADS)

    Rhee, C.-K.; Bae, S. H.; Lee, J. W.; Ahn, J. C.; Jung, J. Y.; Suh, M.-W.

    2009-02-01

    Biofilm formation has been demonstrated for many mucosal pathogens such as Haemophilus influenzae. The presence of mucosal biofilms with chronic otitis media with effusion (COME) suggests that bacteria do not clear by antibiotics. Aim: To test the effect of photodynamic therapy (PDT) on H. influenzae biofilm in vitro. Methods: Sixteen biofilms of H. influenzae were maintained on culture chamber with continuous flow cell system. The biofilms were divided into control, laser, photofrin, and PDT groups. For culture group, the biofilms were cultured. For laser group, 7.2 J/cm2 of 632 nm diode laser was irradiated to the biofilms. For photofrin group, photofrins 5 and 25ug/ml were added to the media. For PDT group, photofrins 5 and 25 ug/ml were added to the media following 632 nm diode laser was irradiated (7.2 J/cm2) to the biofilms. Live/Dead (DAPI/PI) stain was performed and biofilms were examined under confocal laser microscope for thickness and density of biofilms. Results: By DAPI/PI staining, significant reduction of biofilms thickness and complete killing of H. influenzae in PDT group with 25µg photofrin was noted while the biofilms were well maintained in the other groups. Conclusion: The results of this study demonstrated that PDT appears to be effective to photoinactivate experimental H. influenzae biofilms in vitro. Clinical implication: PDT can be a possible alternative treatment to antiobiotic treatment on otitis media with biofilm formation.

  8. Bioreduction of para-chloronitrobenzene in drinking water using a continuous stirred hydrogen-based hollow fiber membrane biofilm reactor.

    PubMed

    Xia, Siqing; Li, Haixiang; Zhang, Zhiqiang; Zhang, Yanhao; Yang, Xin; Jia, Renyong; Xie, Kang; Xu, Xiaotian

    2011-08-30

    para-Chloronitrobenzene (p-CNB) is particularly harmful and persistent in the environment and is one of the priority pollutants. A feasible degradation pathway for p-CNB is bioreduction under anaerobic conditions. Bioreduction of p-CNB using a hydrogen-based hollow fiber membrane biofilm reactor (HFMBfR) was investigated in the present study. The experiment results revealed that p-CNB was firstly reduced to para-chloraniline (p-CAN) as an intermediate and then reduced to aniline that involves nitro reduction and reductive dechlorination with H(2) as the electron donor. The HFMBfR had reduced p-CNB to a major extent with a maximum removal percentage of 99.3% at an influent p-CNB concentration of 2mg/L and a hydraulic residence time of 4.8h, which corresponded to a p-CNB flux of 0.058g/m(2) d. The H(2) availability, p-CNB loading, and the presence of competing electron acceptors affected the p-CNB reduction. Flux analysis indicated that the reduction of p-CNB and p-CAN could consume fewer electrons than that of nitrate and sulfate. The HFMBfR had high average hydrogen utilization efficiencies at different steady states in this experiment, with a maximum efficiency at 98.2%. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Escherichia coli serotype O157:H7 retention on solid surfaces and peroxide resistance is enhanced by dual-strain biofilm formation.

    PubMed

    Uhlich, Gaylen A; Rogers, Donna P; Mosier, Derek A

    2010-08-01

    In a previous study we showed that an Escherichia coli O157:H7 strain that was unable to form biofilm was retained in large numbers in dual-strain biofilms formed with an E. coli O-:H4 companion strain. In this study we tested additional companion strains for their ability to retain E. coli O157:H7 strain 0475s. Companion strains producing biofilm that withstood aggressive washes were able to significantly increase serotype O157:H7 retention. Dual-strain biofilms with certain companion strains retained higher percentages of strain 0475s, and that ability was independent of biofilm total cell numbers. Tests with additional non-biofilm-forming E. coli O157:H7 strains showed that enhancement by companion strains was not unique to strain 0475s. Experiments using an E. coli companion strain with deletions of various curli and cellulose genes indicated that dual-strain biofilm formation was dependent on companion strain properties. Strain 0475s was not able to generate biofilm or persist on plastic when grown in broth with a biofilm-forming companion and separated by a 0.2 microm porous membrane, indicating a requirement for intimate contact with the companion strain. When dual-strain biofilms and planktonic cells were challenged with 5% H(2)O(2), strain 0475 showed greater survival in biofilms with certain companion strains compared to the corresponding planktonic cells. The results of this study indicate that non-biofilm-forming E. coli O157:H7 strains are retained on solid surfaces associated with biofilms generated by companion strains. However, properties other than biofilm mass enable certain companion strains to retain greater numbers of E. coli O157:H7.

  10. Linking composition of extracellular polymeric substances (EPS) to the physical structure and hydraulic resistance of membrane biofilms.

    PubMed

    Desmond, Peter; Best, James P; Morgenroth, Eberhard; Derlon, Nicolas

    2018-04-01

    The effect of extracellular polymeric substances (EPS) on the meso-scale physical structure and hydraulic resistance of membrane biofilms during gravity driven membrane (GDM) filtration was investigated. Biofilms were developed on the surface of ultrafiltration membranes during dead-end filtration at ultra-low pressure (70 mbar). Biofilm EPS composition (total protein, polysaccharide and eDNA) was manipulated by growing biofilms under contrasting nutrient conditions. Nutrient conditions consisted of (i) a nutrient enriched condition with a nutrient ratio of 100:30:10 (C: N: P), (ii) a phosphorus limitation (C: N: P ratio: 100:30:0), and (iii) a nitrogen limitation (C: N: P ratio: 100:0:10). The structure of the biofilm was characterised at meso-scale using Optical Coherence Tomography (OCT). Biofilm composition was analysed with respect to total organic carbon, total cellular mass and extracellular concentrations of proteins, polysaccharides, and eDNA. 2D-confocal Raman mapping was used to characterise the functional group composition and micro-scale distribution of the biofilms EPS. Our study reveals that the composition of the EPS matrix can determine the meso-scale physical structure of membrane biofilms and in turn its hydraulic resistance. Biofilms grown under P limiting conditions were characterised by dense and homogeneous physical structures with high concentrations of polysaccharides and eDNA. Biofilm grown under nutrient enriched or N limiting conditions were characterised by heterogeneous physical structures with lower concentrations of polysaccharides and eDNA. For P limiting biofilms, 2D-confocal Raman microscopy revealed a homogeneous spatial distribution of anionic functional groups in homogeneous biofilm structures with higher polysaccharide and eDNA concentrations. This study links EPS composition, physical structure and hydraulic resistance of membrane biofilms, with practical relevance for the hydraulic performances of GDM ultrafiltration

  11. Determination of pollutant diffusion coefficients in naturally formed biofilms using a single tube extractive membrane bioreactor

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, S.F.; Splendiani, A.; Freitas dos Santos, L.M.

    A novel technique has been used to determine the effective diffusion coefficients for 1,1,2-trichloroethane (TCE), a nonreacting tracer, in biofilms growing on the external surface of a silicone rubber membrane tube during degradation of 1,2-dichloroethane (DCE) by Xanthobacter autotrophicus GJ10 and monochlorobenzene (MCB) by Pseudomonas JS150. Experiments were carried out in a single tube extractive membrane bioreactor (STEMB), whose configuration makes it possible to measure the transmembrane flux of substrates. A video imaging technique (VIT) was employed for in situ biofilm thickness measurement and recording. Diffusion coefficients of TCE in the biofilms and TCE mass transfer coefficients in the liquidmore » films adjacent to the biofilms were determined simultaneously using a resistances-in-series diffusion model. It was found that the flux and overall mass transfer coefficient of TCE decrease with increasing biofilm thickness, showing the importance of biofilm diffusion on the mass transfer process. Similar fluxes were observed for the nonreacting tracer (TCE) and the reactive substrates (MCB or DCE), suggesting that membrane-attached biofilm systems can be rate controlled primarily by substrate diffusion. The TCE diffusion coefficient in the JS150 biofilm appeared to be dependent on biofilm thickness, decreasing markedly for biofilm thicknesses of >1 mm. The values of the TCE diffusion coefficients in the JS150 biofilms <1-mm thick are approximately twice those in water and fall to around 30% of the water value for biofilms >1-mm thick.« less

  12. Quantitative assessment of the efficacy of spiral-wound membrane cleaning procedures to remove biofilms.

    PubMed

    Hijnen, W A M; Castillo, C; Brouwer-Hanzens, A H; Harmsen, D J H; Cornelissen, E R; van der Kooij, D

    2012-12-01

    Cleaning of high pressure RO/NF membranes is an important operational tool to control biofouling. Quantitative information on the efficacy of cleaning agents and protocols to remove biomass is scarce. Therefore, a laboratory cleaning test to assess the efficiency of cleaning procedures to remove attached biomass was developed. The major components of the test are (i) production of uniform biofilm samples, (ii) the quantification of the biomass concentrations with robust parameters and (iii) a simple test procedure with optimal exposure of the biofilm samples to the chemicals. The results showed that PVC-P is a suitable substratum for the production of uniform biofilm samples. ATP and carbohydrates (CH) as major components of the biofilm matrix for nucleotides (living bacterial cells) and extracellular polymeric substances EPS, respectively, were selected as robust biomass parameters. The removal of ATP and CH with the NaOH/Sodium Dodecyl Sulfate (SDS) mixture, selected as a standard treatment at pH 12.0, was reproducible. The resistance of the EPS matrix against chemical cleaning was demonstrated by a low CH removal (32.8 ± 6.0%) compared to the ATP removal (70.5 ± 15.1%). The inverse relationship of biomass removal with the CH to ATP ratio (μg/ng) of the biofilms demonstrated the influence of the biomass characteristics on cleaning. None of the 27 chemicals tested (analytical-grade and commercial brands) in single step or in double-step treatments were significantly more effective than NaOH/SDS. Oxidizing agents NaOCl and H(2)O(2), the latter in combination with SDS, both tested as common agents in biofilm control, showed a significantly higher efficiency (70%) to remove biofilms. In the test, simultaneously, the efficiency of agents to remove precipitated minerals such as Fe can be assessed. Validation tests with Cleaning in Place (CIP) in 8 and 2.5-inch RO membrane pilot plant experiments showed similar ranking of the cleaning efficiency of cleaning protocols

  13. Biofouling of reverse osmosis membranes: effects of cleaning on biofilm microbial communities, membrane performance, and adherence of extracellular polymeric substances.

    PubMed

    Al Ashhab, Ashraf; Sweity, Amer; Bayramoglu, Bihter; Herzberg, Moshe; Gillor, Osnat

    2017-05-01

    Laboratory-scale reverse osmosis (RO) flat-sheet systems were used with two parallel flow cells, one treated with cleaning agents and a control (ie undisturbed). The cleaning efforts increased the affinity of extracellular polymeric substances (EPS) to the RO membrane and altered the biofilm surface structure. Analysis of the membrane biofilm community composition revealed the dominance of Proteobacteria. However, within the phylum Proteobacteria, γ-Proteobacteria dominated the cleaned membrane biofilm, while β-Proteobacteria dominated the control biofilm. The composition of the fungal phyla was also altered by cleaning, with enhancement of Ascomycota and suppression of Basidiomycota. The results suggest that repeated cleaning cycles select for microbial groups that strongly attach to the RO membrane surface by producing rigid and adhesive EPS that hampers membrane performance.

  14. Effect of engineered environment on microbial community structure in biofilter and biofilm on reverse osmosis membrane.

    PubMed

    Jeong, Sanghyun; Cho, Kyungjin; Jeong, Dawoon; Lee, Seockheon; Leiknes, TorOve; Vigneswaran, Saravanamuthu; Bae, Hyokwan

    2017-11-01

    Four dual media filters (DMFs) were operated in a biofiltration mode with different engineered environments (DMF I and II: coagulation with/without acidification and DMF III and IV: without/with chlorination). Designed biofilm enrichment reactors (BERs) containing the removable reverse osmosis (RO) coupons, were connected at the end of the DMFs in parallel to analyze the biofilm on the RO membrane by DMF effluents. Filtration performances were evaluated in terms of dissolved organic carbon (DOC) and assimilable organic carbon (AOC). Organic foulants on the RO membrane were also quantified and fractionized. The bacterial community structures in liquid (seawater and effluent) and biofilm (DMF and RO) samples were analyzed using 454-pyrosequencing. The DMF IV fed with the chlorinated seawater demonstrated the highest reductions of DOC including LMW-N as well as AOC among the other DMFs. The DMF IV was also effective in reducing organic foulants on the RO membrane surface. The bacterial community structure was grouped according to the sample phase (i.e., liquid and biofilm samples), sampling location (i.e., DMF and RO samples), and chlorination (chlorinated and non-chlorinated samples). In particular, the biofilm community in the DMF IV differed from the other DMF treatments, suggesting that chlorination exerted as stronger selective pressure than pH adjustment or coagulation on the biofilm community. In the DMF IV, several chemoorganotrophic chlorine-resistant biofilm-forming bacteria such as Hyphomonas, Erythrobacter, and Sphingomonas were predominant, and they may enhance organic carbon degradation efficiency. Diverse halophilic or halotolerant organic degraders were also found in other DMFs (i.e., DMF I, II, and III). Various kinds of dominant biofilm-forming bacteria were also investigated in RO membrane samples; the results provided possible candidates that cause biofouling when DMF process is applied as the pretreatment option for the RO process. Copyright

  15. pH, redox potential and local biofilm potential microenvironments within Geobacter sulfurreducens biofilms and their roles in electron transfer.

    PubMed

    Babauta, Jerome T; Nguyen, Hung Duc; Harrington, Timothy D; Renslow, Ryan; Beyenal, Haluk

    2012-10-01

    The limitation of pH inside electrode-respiring biofilms is a well-known concept. However, little is known about how pH and redox potential are affected by increasing current inside biofilms respiring on electrodes. Quantifying the variations in pH and redox potential with increasing current is needed to determine how electron transfer is tied to proton transfer within the biofilm. In this research, we quantified pH and redox potential variations in electrode-respiring Geobacter sulfurreducens biofilms as a function of respiration rates, measured as current. We also characterized pH and redox potential at the counter electrode. We concluded that (1) pH continued to decrease in the biofilm through different growth phases, showing that the pH is not always a limiting factor in a biofilm and (2) decreasing pH and increasing redox potential at the biofilm electrode were associated only with the biofilm, demonstrating that G. sulfurreducens biofilms respire in a unique internal environment. Redox potential inside the biofilm was also compared to the local biofilm potential measured by a graphite microelectrode, where the tip of the microelectrode was allowed to acclimatize inside the biofilm. Copyright © 2012 Wiley Periodicals, Inc.

  16. The effect of PDT on H. influenzae biofilm in vivo

    NASA Astrophysics Data System (ADS)

    Rhee, Chung-Ku; Chang, So-Young; Hwang, Dong-Jo; Kim, Young Hoon; Ahn, Jin-Chul

    2010-02-01

    Biofilm formation has been demonstrated for many mucosal pathogens such as Haemophilus influenzae. The presence of mucosal biofilms with chronic otitis media with effusion (COME) suggests that bacteria do not clear by antibiotics. Aim: To test the effect of photodynamic therapy (PDT) on H. influenzae induced biofilm in vivo. Methods: Gerbils were divided into control (C), HI group, Laser (L), PS, PDT A, and PDT B groups. The C group received no treatment. HI group was injected with 20μl (108CFU/ml) of H. influenzae into the bullae and formation of biofilms in the bullae was obtained by 5 days. For L group, 120 J/cm2 (100 mw × 20 min) of 632 nm LD laser was irradiated by a fiber inserted into the bullae 5 days after the H. influenzaeinjection. For PS group, photofrin 40μl (1mg/ml) were injected into the bullae 5 days after the H. influenzae injection. PDT A group received photofrin 1 mg/ml and LD laser 120 J/cm2 that were administered into the bullae 5 days after the H. influenzae injection. PDT B group received photofrin 2 mg/ml and laser 150 J/cm2 5 days after the H. influenzae injection. The mucosal tissues in bullae were examined by H/E staining, and SEM. Results: The C group showed normal mucosa of bullae. The HI, L, and PS groups have shown well formed biofilm. Twenty five percent of the PDT A group and 50 % of the PDT B group have shown completely or partially resolved biofilm. Conclusion: The results of this study demonstrated that PDT appears to be effective to treat experimental H. influenzae induced biofilms in vivo. Clinical implication: PDT may be an alternative to antibiotic treatment on otitis media with biofilm formation.

  17. Membrane distillation combined with an anaerobic moving bed biofilm reactor for treating municipal wastewater.

    PubMed

    Kim, Hyun-Chul; Shin, Jaewon; Won, Seyeon; Lee, Jung-Yeol; Maeng, Sung Kyu; Song, Kyung Guen

    2015-03-15

    A fermentative strategy with an anaerobic moving bed biofilm reactor (AMBBR) was used for the treatment of domestic wastewater. The feasibility of using a membrane separation technique for post-treatment of anaerobic bio-effluent was evaluated with emphasis on employing a membrane distillation (MD). Three different hydrophobic 0.2 μm membranes made of polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), and polypropylene (PP) were examined in this study. The initial permeate flux of the membranes ranged from 2.5 to 6.3 L m(-2h(-1) when treating AMBBR effluent at a temperature difference between the feed and permeate streams of 20 °C, with the permeate flux increasing in the order PP < PVDF < PTFE. The permeate flux of the PTFE membrane gradually decreased to 84% of the initial flux after the 45 h run for distillation, while a flux decline in MD with either the PVDF or PP membrane was not found under the identical distillation conditions. During long-term distillation with the PVDF membrane, total phosphorus was completely rejected and >98% rejection of dissolved organic carbon was also achieved. The characterization of wastewater effluent organic matter (EfOM) using an innovative suite of analytical tools verified that almost all of the EfOM was rejected via the PVDF MD treatment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Performances of biological aerated filter employing hollow fiber membrane segments of surface-improved poly (sulfone) as biofilm carriers.

    PubMed

    Shen, Ying-Jie; Wu, Guang-Xia; Fan, Yao-Bo; Zhong, Hui; Wu, Lin-Lin; Zhang, Shao-Lai; Zhao, Xian-Hong; Zhang, Wei-Jun

    2007-01-01

    Using the surface of poly (sulfone) hollow fiber membrane segments as grafted layer, the hydrophilic acrylamide chain was grafted on by UV-photoinduced grafting polymerization. The gained improvement of surface wettability for the modified membrane was tested by measuring the contact-angle as well as FTIR spectra. Then correlation between the hydrophilic ability of support material and the biofilm adherence ability was demonstrated by comparing the pollutant removal rates from urban wastewater via two identical lab-scale up-flow biological aerated filters, one employed the surface wettability modified poly (sulfone) hollow fiber membrane segment as biofilm carrier and the other employed unmodified membrane segment as biofilm carrier. The experimental results showed that under the conditions of influent flux 5 L/h, hydraulic retention time 9 h and gas to liquid ratio (G/L) 10:1, the removal rates of chemical oxygen demand (COD) and ammonium nitrogen (NH4(+)-N) for the modified packing filter and the unmodified packing filter was averaged at 83.64% and 96.25%, respectively, with the former filter being 5%-20% more than the latter. The effluent concentration of COD, NH4(+)-N and turbidity for the modified packing filter was 25.25 mg/L, 2 mg/L and 8 NTU, respectively. Moreover, the ammonium nitrogen removal performance of the filter packing the modified PSF was compared with the other bioreactor packing of an efficient floating medium. The biomass test indicated that the modified membrane matrixes provided better specific adhesion (3310-5653 mg TSS/L support), which gave a mean of 1000 mg TSS/L more than the unmodified membrane did. In addition, the phenomenon of simultaneous denitrification on the inner surface of the support and nitrification on the outer surface was found in this work.

  19. Helicobacter pylori ATCC 43629/NCTC 11639 Outer Membrane Vesicles (OMVs) from Biofilm and Planktonic Phase Associated with Extracellular DNA (eDNA)

    PubMed Central

    Grande, Rossella; Di Marcantonio, Maria C.; Robuffo, Iole; Pompilio, Arianna; Celia, Christian; Di Marzio, Luisa; Paolino, Donatella; Codagnone, Marilina; Muraro, Raffaella; Stoodley, Paul; Hall-Stoodley, Luanne; Mincione, Gabriella

    2015-01-01

    Helicobacter pylori persistence is associated with its capacity to develop biofilms as a response to changing environmental conditions and stress. Extracellular DNA (eDNA) is a component of H. pylori biofilm matrix but the lack of DNase I activity supports the hypothesis that eDNA might be protected by other extracellular polymeric substances (EPS) and/or Outer Membrane Vesicles (OMVs), which bleb from the bacteria surface during growth. The aim of the present study was to both identify the eDNA presence on OMVs segregated from H. pylori ATCC 43629/NCTC 11639 biofilm (bOMVs) and its planktonic phase (pOMVs) and to characterize the physical-chemical properties of the OMVs. The presence of eDNA in bOMVs and pOMVs was initially carried out using DNase I-gold complex labeling and Transmission Electron Microscope analysis (TEM). bOMVs and pOMVs were further isolated and physical-chemical characterization carried out using dynamic light scattering (DLS) analysis. eDNA associated with OMVs was detected and quantified using a PicoGreen spectrophotometer assay, while its extraction was performed with a DNA Kit. TEM images showed that eDNA was mainly associated with the OMV membrane surfaces; while PicoGreen staining showed a four-fold increase of dsDNA in bOMVs compared with pOMVs. The eDNA extracted from OMVs was visualized using gel electrophoresis. DLS analysis indicated that both planktonic and biofilm H. pylori phenotypes generated vesicles, with a broad distribution of sizes on the nanometer scale. The DLS aggregation assay suggested that eDNA may play a role in the aggregation of OMVs, in the biofilm phenotype. Moreover, the eDNA associated with vesicle membrane may impede DNase I activity on H. pylori biofilms. These results suggest that OMVs derived from the H. pylori biofilm phenotype may play a structural role by preventing eDNA degradation by nucleases, providing a bridging function between eDNA strands on OMV surfaces and promoting aggregation. PMID:26733944

  20. Zeolitic Imidazolate Framework-8 Membrane for H2/CO2 Separation: Experimental and Modeling

    NASA Astrophysics Data System (ADS)

    Lai, L. S.; Yeong, Y. F.; Lau, K. K.; Azmi, M. S.; Chew, T. L.

    2018-03-01

    In this work, ZIF-8 membrane synthesized through solvent evaporation secondary seeded growth was tested for single gas permeation and binary gases separation of H2 and CO2. Subsequently, a modified mathematical modeling combining the effects of membrane and support layers was applied to represent the gas transport properties of ZIF-8 membrane. Results showed that, the membrane has exhibited H2/CO2 ideal selectivity of 5.83 and separation factor of 3.28 at 100 kPa and 303 K. Besides, the experimental results were fitted well with the simulated results by demonstrating means absolute error (MAE) values ranged from 1.13 % to 3.88 % for single gas permeation and 10.81 % to 21.22 % for binary gases separation. Based on the simulated data, most of the H2 and CO2 gas molecules have transported through the molecular pores of membrane layer, which was up to 70 %. Thus, the gas transport of the gases is mainly dominated by adsorption and diffusion across the membrane.

  1. Maltodextrin enhances biofilm elimination by electrochemical scaffold

    PubMed Central

    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

  2. Effect of stabilization temperature during pyrolysis process of P84 co-polyimide-based tubular carbon membrane for H2/N2 and He/N2 separations

    NASA Astrophysics Data System (ADS)

    Sazali, N.; Salleh, W. N. W.; Ismail, A. F.; Ismail, N. H.; Aziz, F.; Yusof, N.; Hasbullah, H.

    2018-04-01

    In this study, the effect of stabilization temperature on the performance of tubular carbon membrane was being investigated. P84 co-polyimide-based tubular carbon membrane will be fabricated through the dip-coating technique. The tubular carbon membrane performance can be controlled by manipulating the pyrolysis conditions which was conducted at different stabilization temperatures of 250, 300, 350, 400, and 450°C under N2 environment (200 ml/min). The prepared membranes were characterized by using scanning electron microscopy (SEM), x-ray diffraction (XRD), and pure gas permeation system. The pure gas of H2, He, and N2 were used to determine the permeation properties of the carbon membrane. The P84 co-polyimide-based tubular carbon membrane stabilized at 300°C demonstrated an excellent permeation property with H2, He, and N2 gas permeance of 1134.51±2.87, 1287.22±2.86 and 2.98±1.28GPU, respectively. The highest H2/N2 and He/N2 selectivity of 380.71±2.34 and 431.95±2.61 was obtained when the stabilization temperature of 450°C was applied. It is concluded that the stabilization temperatures have protrusive effect on the carbon membrane properties specifically their pore structure, and eventually their gas separation properties.

  3. The Vibrio cholerae Pst2 phosphate transport system is upregulated in biofilms and contributes to biofilm-induced hyperinfectivity.

    PubMed

    Mudrak, Benjamin; Tamayo, Rita

    2012-05-01

    Vibrio cholerae is the causative agent of the deadly diarrheal disease cholera. As part of its life cycle, V. cholerae persists in marine environments, where it forms surface-attached communities commonly described as biofilms. Evidence indicates that these biofilms constitute the infectious form of the pathogen during outbreaks. Previous work has shown that biofilm-derived V. cholerae cells, even when fully dispersed from the biofilm matrix, are vastly more infectious than planktonic (free-living) cells. Here, we sought to identify factors that contribute to biofilm-induced hyperinfectivity in V. cholerae, and we present evidence for one aspect of the molecular basis of this phenotype. We identified proteins upregulated during growth in biofilms and determined their contributions to the hyperinfectivity phenotype. We found that PstS2, the periplasmic component of the Pst2 phosphate uptake system, was enriched in biofilms. Another gene in the pst2 locus was transcriptionally upregulated in biofilms. Using the infant mouse model, we found that mutation of two pst2 components resulted in impaired colonization. Importantly, deletion of the Pst2 inner membrane complex caused a greater colonization defect after growth in a biofilm compared to shaking culture. Based on these data, we propose that V. cholerae cells in biofilms upregulate the Pst2 system and therefore gain an advantage upon entry into the host. Further characterization of factors contributing to biofilm-induced hyperinfectivity in V. cholerae will improve our understanding of the transmission of the bacteria from natural aquatic habitats to the human host.

  4. Synergistic bactericidal action of phytic acid and sodium chloride against Escherichia coli O157:H7 cells protected by a biofilm.

    PubMed

    Kim, Nam Hee; Rhee, Min Suk

    2016-06-16

    The food industry must prevent the build-up of strong Escherichia coli O157:H7 biofilms in food processing environments. The present study examined the bactericidal action of phytic acid (PA), a natural extract from rice bran and the hulls/peels of legumes, against E. coli O157:H7 biofilms. The synergistic bactericidal effects of PA plus sodium chloride (NaCl) were also examined. E. coli O157:H7 biofilms were allowed for form on stainless steel coupons by culture in both rich (tryptic soy broth, TSB) and minimal (M9) medium at 22°C for 6days. Bacterial cells within biofilms grown in M9 medium were significantly more resistant to PA than those grown in TSB (p<0.05); thus M9 medium was selected for further experiments. The anti-biofilm effect of PA was significantly increased by addition of NaCl (2-4%) (p<0.05); indeed, the combination of 0.4% PA plus 3-4% NaCl completely inactivated E. coli O157:H7 biofilms without recovery (a>6.5logCFU/cm(2) reduction). Neither PA nor NaCl alone were this effective (PA, 1.6-2.7logCFU/cm(2) reduction; NaCl, <0.5logCFU/cm(2) reduction). Confocal laser scanning microscopy images of propidium iodide-treated cells showed that PA (0.4%) plus NaCl (2-4%) had marked membrane permeabilizing effects. These results suggest that a sanitizer that combines these two naturally occurring antimicrobial agents may be useful to food safety managers who encounter thick biofilm formation in food processing environments. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Biofilm Filtrates of Pseudomonas aeruginosa Strains Isolated from Cystic Fibrosis Patients Inhibit Preformed Aspergillus fumigatus Biofilms via Apoptosis

    PubMed Central

    Shirazi, Fazal; Ferreira, Jose A. G.; Stevens, David A.; Clemons, Karl V.; Kontoyiannis, Dimitrios P.

    2016-01-01

    Pseudomonas aeruginosa (Pa) and Aspergillus fumigatus (Af) colonize cystic fibrosis (CF) patient airways. Pa culture filtrates inhibit Af biofilms, and Pa non-CF, mucoid (Muc-CF) and nonmucoid CF (NMuc-CF) isolates form an ascending inhibitory hierarchy. We hypothesized this activity is mediated through apoptosis induction. One Af and three Pa (non-CF, Muc-CF, NMuc-CF) reference isolates were studied. Af biofilm was formed in 96 well plates for 16 h ± Pa biofilm filtrates. After 24 h, apoptosis was characterized by viability dye DiBAc, reactive oxygen species (ROS) generation, mitochondrial membrane depolarization, DNA fragmentation and metacaspase activity. Muc-CF and NMuc-CF filtrates inhibited and damaged Af biofilm (p<0.0001). Intracellular ROS levels were elevated (p<0.001) in NMuc-CF-treated Af biofilms (3.7- fold) compared to treatment with filtrates from Muc-CF- (2.5- fold) or non-CF Pa (1.7- fold). Depolarization of mitochondrial potential was greater upon exposure to NMuc-CF (2.4-fold) compared to Muc-CF (1.8-fold) or non-CF (1.25-fold) (p<0.0001) filtrates. Exposure to filtrates resulted in more DNA fragmentation in Af biofilm, compared to control, mediated by metacaspase activation. In conclusion, filtrates from CF-Pa isolates were more inhibitory against Af biofilms than from non-CF. The apoptotic effect involves mitochondrial membrane damage associated with metacaspase activation. PMID:26930399

  6. Biofilm Filtrates of Pseudomonas aeruginosa Strains Isolated from Cystic Fibrosis Patients Inhibit Preformed Aspergillus fumigatus Biofilms via Apoptosis.

    PubMed

    Shirazi, Fazal; Ferreira, Jose A G; Stevens, David A; Clemons, Karl V; Kontoyiannis, Dimitrios P

    2016-01-01

    Pseudomonas aeruginosa (Pa) and Aspergillus fumigatus (Af) colonize cystic fibrosis (CF) patient airways. Pa culture filtrates inhibit Af biofilms, and Pa non-CF, mucoid (Muc-CF) and nonmucoid CF (NMuc-CF) isolates form an ascending inhibitory hierarchy. We hypothesized this activity is mediated through apoptosis induction. One Af and three Pa (non-CF, Muc-CF, NMuc-CF) reference isolates were studied. Af biofilm was formed in 96 well plates for 16 h ± Pa biofilm filtrates. After 24 h, apoptosis was characterized by viability dye DiBAc, reactive oxygen species (ROS) generation, mitochondrial membrane depolarization, DNA fragmentation and metacaspase activity. Muc-CF and NMuc-CF filtrates inhibited and damaged Af biofilm (p<0.0001). Intracellular ROS levels were elevated (p<0.001) in NMuc-CF-treated Af biofilms (3.7- fold) compared to treatment with filtrates from Muc-CF- (2.5- fold) or non-CF Pa (1.7- fold). Depolarization of mitochondrial potential was greater upon exposure to NMuc-CF (2.4-fold) compared to Muc-CF (1.8-fold) or non-CF (1.25-fold) (p<0.0001) filtrates. Exposure to filtrates resulted in more DNA fragmentation in Af biofilm, compared to control, mediated by metacaspase activation. In conclusion, filtrates from CF-Pa isolates were more inhibitory against Af biofilms than from non-CF. The apoptotic effect involves mitochondrial membrane damage associated with metacaspase activation.

  7. Ratiometric Imaging of Extracellular pH in Dental Biofilms.

    PubMed

    Schlafer, Sebastian; Dige, Irene

    2016-03-09

    The pH in bacterial biofilms on teeth is of central importance for dental caries, a disease with a high worldwide prevalence. Nutrients and metabolites are not distributed evenly in dental biofilms. A complex interplay of sorption to and reaction with organic matter in the biofilm reduces the diffusion paths of solutes and creates steep gradients of reactive molecules, including organic acids, across the biofilm. Quantitative fluorescent microscopic methods, such as fluorescence life time imaging or pH ratiometry, can be employed to visualize pH in different microenvironments of dental biofilms. pH ratiometry exploits a pH-dependent shift in the fluorescent emission of pH-sensitive dyes. Calculation of the emission ratio at two different wavelengths allows determining local pH in microscopic images, irrespective of the concentration of the dye. Contrary to microelectrodes the technique allows monitoring both vertical and horizontal pH gradients in real-time without mechanically disturbing the biofilm. However, care must be taken to differentiate accurately between extra- and intracellular compartments of the biofilm. Here, the ratiometric dye, seminaphthorhodafluor-4F 5-(and-6) carboxylic acid (C-SNARF-4) is employed to monitor extracellular pH in in vivo grown dental biofilms of unknown species composition. Upon exposure to glucose the dye is up-concentrated inside all bacterial cells in the biofilms; it is thus used both as a universal bacterial stain and as a marker of extracellular pH. After confocal microscopic image acquisition, the bacterial biomass is removed from all pictures using digital image analysis software, which permits to exclusively calculate extracellular pH. pH ratiometry with the ratiometric dye is well-suited to study extracellular pH in thin biofilms of up to 75 µm thickness, but is limited to the pH range between 4.5 and 7.0.

  8. Performance of a composite membrane bioreactor treating toluene vapors: inocula selection, reactor performance and behavior under transient conditions.

    PubMed

    Kumar, Amit; Dewulf, Jo; Vercruyssen, Aline; Van Langenhove, Herman

    2009-04-01

    In this study, a membrane biofilm reactor performance for toluene as a model pollutant is presented. A composite membrane consisting of a porous polyacrylonitrile (PAN) support layer coated with a very thin (0.3 microm) dense polydimethylsiloxane (PDMS) top layer was used. Batch experiments were performed to select an appropriate inocula (slaughterhouse wastewater treatment sludge with a specific toluene consumption rate of 118+/-23 microg g(-1) VSS L(-1)) among the three available sources of inoculums. The maximum elimination capacity gas-side reactor volume based (EC)v and membrane based (EC)(m, max) obtained were 609 g m(-3) h(-1) and 1.2 g m(-2) h(-1) respectively, which is much higher than other membrane bioreactors. Further experiments involved the study of the membrane biofilm reactor flexibility when operational parameters as temperature, loading rate etc. were modified. In all cases, the membrane biofilm reactor showed a rapid adaptation and new steady-states were obtained within hours. Overall, the results illustrate that membrane bioreactors can potentially be a good option for treatment of air pollutants such as toluene.

  9. Perchlorate Destruction and Potable Water Production Using Membrane Biofilm Reduction and Membrane Filtration

    DTIC Science & Technology

    2013-11-18

    the experimental filter media Next-SandTM was used, thus turbidity results may not be translatable to conventional filtration media. The media...performance objective was not met. Further optimization of the media filtration process would result in meeting the objective. Dissolved Organic Carbon...FINAL REPORT Perchlorate Destruction and Potable Water Production Using Membrane Biofilm Reduction and Membrane Filtration ESTCP Project ER

  10. Start-up of membrane bioreactor and hybrid moving bed biofilm reactor-membrane bioreactor: kinetic study.

    PubMed

    Leyva-Díaz, J C; Poyatos, J M

    2015-01-01

    A hybrid moving bed biofilm reactor-membrane bioreactor (hybrid MBBR-MBR) system was studied as an alternative solution to conventional activated sludge processes and membrane bioreactors. This paper shows the results obtained from three laboratory-scale wastewater treatment plants working in parallel in the start-up and steady states. The first wastewater treatment plant was a MBR, the second one was a hybrid MBBR-MBR system containing carriers both in anoxic and aerobic zones of the bioreactor (hybrid MBBR-MBRa), and the last one was a hybrid MBBR-MBR system which contained carriers only in the aerobic zone (hybrid MBBR-MBRb). The reactors operated with a hydraulic retention time of 30.40 h. A kinetic study for characterizing heterotrophic biomass was carried out and organic matter and nutrients removals were evaluated. The heterotrophic biomass of the hybrid MBBR-MBRb showed the best kinetic performance in the steady state, with yield coefficient for heterotrophic biomass=0.30246 mg volatile suspended solids per mg chemical oxygen demand, maximum specific growth rate for heterotrophic biomass=0.00308 h(-1) and half-saturation coefficient for organic matter=3.54908 mg O2 L(-1). The removal of organic matter was supported by the kinetic study of heterotrophic biomass.

  11. Simultaneous Bioreduction of Multiple Oxidized Contaminants Using a Membrane Biofilm Reactor.

    PubMed

    Li, Haixiang; Lin, Hua; Xu, Xiaoyin; Jiang, Minmin; Chang, Chein-Chi; Xia, Siqing

    2017-02-01

      This study tests a hydrogen-based membrane biofilm reactor (MBfR) to investigate simultaneous bioreduction of selected oxidized contaminants, including nitrate (-N), sulfate (), bromate (), chromate (Cr(VI)) and para-chloronitrobenzene (p-CNB). The experiments demonstrate that MBfR can achieve high performance for contaminants bioreduction to harmless or immobile forms in 240 days, with a maximum reduction fluxes of 0.901 g -N/m2·d, 1.573 g /m2·d, 0.009 g /m2·d, 0.022 g Cr(VI)/m2·d, and 0.043 g p-CNB/m2·d. Increasing H2 pressure and decreasing influent surface loading enhanced removal efficiency of the reactor. Flux analysis indicates that nitrate and sulfate reductions competed more strongly than , Cr(VI) and p-CNB reduction. The average H2 utilization rate, H2 flux, and H2 utilization efficiency of the reactor were 0.026 to 0.052 mg H2/cm3·d, 0.024 to 0.046 mg H2/cm2·d, and 97.5% to 99.3% (nearly 100%). Results show the hydrogen-based MBfR may be suitable for removing multiple oxidized contaminants in drinking water or groundwater.

  12. Two-step nitrification in a pure moving bed biofilm reactor-membrane bioreactor for wastewater treatment: nitrifying and denitrifying microbial populations and kinetic modeling.

    PubMed

    Leyva-Díaz, J C; González-Martínez, A; Muñío, M M; Poyatos, J M

    2015-12-01

    The moving bed biofilm reactor-membrane bioreactor (MBBR-MBR) is a novel solution to conventional activated sludge processes and membrane bioreactors. In this study, a pure MBBR-MBR was studied. The pure MBBR-MBR mainly had attached biomass. The bioreactor operated with a hydraulic retention time (HRT) of 9.5 h. The kinetic parameters for heterotrophic and autotrophic biomasses, mainly nitrite-oxidizing bacteria (NOB), were evaluated. The analysis of the bacterial community structure of the ammonium-oxidizing bacteria (AOB), NOB, and denitrifying bacteria (DeNB) from the pure MBBR-MBR was carried out by means of pyrosequencing to detect and quantify the contribution of the nitrifying and denitrifying bacteria in the total bacterial community. The relative abundance of AOB, NOB, and DeNB were 5, 1, and 3%, respectively, in the mixed liquor suspended solids (MLSS), and these percentages were 18, 5, and 2%, respectively, in the biofilm density (BD) attached to carriers. The pure MBBR-MBR had a high efficiency of total nitrogen (TN) removal of 71.81±16.04%, which could reside in the different bacterial assemblages in the fixed biofilm on the carriers. In this regard, the kinetic parameters for autotrophic biomass had values of YA=2.3465 mg O2 mg N(-1), μm, A=0.7169 h(-1), and KNH=2.0748 mg NL(-1).

  13. Structural determinants of Actinomyces sortase SrtC2 required for membrane localization and assembly of type 2 fimbriae for interbacterial coaggregation and oral biofilm formation.

    PubMed

    Wu, Chenggang; Mishra, Arunima; Reardon, Melissa E; Huang, I-Hsiu; Counts, Sarah C; Das, Asis; Ton-That, Hung

    2012-05-01

    As a pioneer colonizer of the oral cavity, Actinomyces oris expresses proteinaceous pili (also called fimbriae) to mediate the following two key events in biofilm formation: adherence to saliva deposits on enamel and interbacterial associations. Assembly of type 2 fimbriae that directly facilitate coaggregation with oral streptococci and Actinomyces biofilm development requires the class C sortase SrtC2. Although the general sortase-associated mechanisms have been elucidated, several structural attributes unique to the class C sortases require functional investigation. Mutational studies reported here suggest that the N-terminal transmembrane (TM) region of SrtC2, predicted to contain a signal peptide sequence, is cleaved off the mature protein and that this processing is critical for the proper integration of the enzyme at the cytoplasmic membrane, which is mediated by the extended hydrophobic C terminus containing a TM domain and a cytoplasmic tail. Deletion of this putative TM or the entire cytoplasmic domain abolished the enzyme localization and functionality. Alanine substitution of the conserved catalytic Cys-His dyad abrogated the SrtC2 enzymatic activity. In contrast, mutations designed to alter a "lid" domain that covers the catalytic pocket of a class C sortase showed no effect on enzyme activity. Finally, each of the deleterious mutations that affected SrtC2 activity or membrane localization also eliminated Actinomyces species biofilm development and bacterial coaggregation with streptococci. We conclude that the N terminus of SrtC2, which contains the signal sequence, is required for proper protein translocation and maturation, while the extended C-terminal hydrophobic region serves as a stable membrane anchor for proper enzyme functionality.

  14. Structural Determinants of Actinomyces sortase SrtC2 Required for Membrane Localization and Assembly of Type 2 Fimbriae for Interbacterial Coaggregation and Oral Biofilm Formation

    PubMed Central

    Wu, Chenggang; Mishra, Arunima; Reardon, Melissa E.; Huang, I-Hsiu; Counts, Sarah C.; Das, Asis

    2012-01-01

    As a pioneer colonizer of the oral cavity, Actinomyces oris expresses proteinaceous pili (also called fimbriae) to mediate the following two key events in biofilm formation: adherence to saliva deposits on enamel and interbacterial associations. Assembly of type 2 fimbriae that directly facilitate coaggregation with oral streptococci and Actinomyces biofilm development requires the class C sortase SrtC2. Although the general sortase-associated mechanisms have been elucidated, several structural attributes unique to the class C sortases require functional investigation. Mutational studies reported here suggest that the N-terminal transmembrane (TM) region of SrtC2, predicted to contain a signal peptide sequence, is cleaved off the mature protein and that this processing is critical for the proper integration of the enzyme at the cytoplasmic membrane, which is mediated by the extended hydrophobic C terminus containing a TM domain and a cytoplasmic tail. Deletion of this putative TM or the entire cytoplasmic domain abolished the enzyme localization and functionality. Alanine substitution of the conserved catalytic Cys-His dyad abrogated the SrtC2 enzymatic activity. In contrast, mutations designed to alter a “lid” domain that covers the catalytic pocket of a class C sortase showed no effect on enzyme activity. Finally, each of the deleterious mutations that affected SrtC2 activity or membrane localization also eliminated Actinomyces species biofilm development and bacterial coaggregation with streptococci. We conclude that the N terminus of SrtC2, which contains the signal sequence, is required for proper protein translocation and maturation, while the extended C-terminal hydrophobic region serves as a stable membrane anchor for proper enzyme functionality. PMID:22447896

  15. Modeling and predicting the biofilm formation of Salmonella Virchow with respect to temperature and pH.

    PubMed

    Ariafar, M Nima; Buzrul, Sencer; Akçelik, Nefise

    2016-03-01

    Biofilm formation of Salmonella Virchow was monitored with respect to time at three different temperature (20, 25 and 27.5 °C) and pH (5.2, 5.9 and 6.6) values. As the temperature increased at a constant pH level, biofilm formation decreased while as the pH level increased at a constant temperature, biofilm formation increased. Modified Gompertz equation with high adjusted determination coefficient (Radj(2)) and low mean square error (MSE) values produced reasonable fits for the biofilm formation under all conditions. Parameters of the modified Gompertz equation could be described in terms of temperature and pH by use of a second order polynomial function. In general, as temperature increased maximum biofilm quantity, maximum biofilm formation rate and time of acceleration of biofilm formation decreased; whereas, as pH increased; maximum biofilm quantity, maximum biofilm formation rate and time of acceleration of biofilm formation increased. Two temperature (23 and 26 °C) and pH (5.3 and 6.3) values were used up to 24 h to predict the biofilm formation of S. Virchow. Although the predictions did not perfectly match with the data, reasonable estimates were obtained. In principle, modeling and predicting the biofilm formation of different microorganisms on different surfaces under various conditions could be possible.

  16. Reduction of Escherichia coli O157:H7 in Biofilms Using Bacteriophage BPECO 19.

    PubMed

    Sadekuzzaman, Mohammad; Yang, Sungdae; Mizan, Md Furkanur Rahaman; Ha, Sang-Do

    2017-06-01

    Biofilm formation is a growing concern in the food industry. Escherichia coli O157:H7 is one of the most important foodborne pathogens that can persists in food and food-related environments and subsequently produce biofilms. The efficacy of bacteriophage BPECO 19 was evaluated against three E. coli O157:H7 strains in biofilms. Biofilms of the three E. coli O157:H7 strains were grown on abiotic (stainless steel, rubber, and minimum biofilm eradication concentration [MBEC TM ] device) and biotic (lettuce) surfaces at different temperatures. The effectiveness of bacteriophage BPECO 19 in reducing preformed biofilms on these surfaces was further evaluated by treating the surfaces with a phage suspension (10 8 PFU/mL) for 2 h. The results indicated that the phage treatment significantly reduced (P  < 0.05) the number of adhered cells in all the surfaces. Following phage treatment, the viability of adhered cells was reduced by ≥3 log CFU/cm 2 , 2.4 log CFU/cm 2 , and 3.1 log CFU/peg in biofilms grown on stainless steel, rubber, and the MBEC TM device, respectively. Likewise, the phage treatment reduced cell viability by ≥2 log CFU/cm 2 in biofilms grown on lettuce. Overall, these results suggested that bacteriophages such as BPECO 19 could be effective in reducing the viability of biofilm-adhered cells. © 2017 Institute of Food Technologists®.

  17. Heterotrophic Archaea Contribute to Carbon Cycling in Low-pH, Suboxic Biofilm Communities

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Justice, Nicholas B; Pan, Chongle; Mueller, Ryan

    Archaea are widely distributed and yet are most often not the most abundant members of microbial communities. Here, we document a transition from Bacteria- to Archaea-dominated communities in microbial biofilms sampled from the Richmond Mine acid mine drainage (AMD) system (pH 1.0,38 C) and in laboratory-cultivated biofilms. This transition occurs when chemoautotrophic microbial communities that develop at the air-solution interface sink to the sediment-solution interface and degrade under microaerobic and anaerobic conditions. The archaea identified in these sunken biofilms are from the class Thermoplasmata, and in some cases, the highly divergent ARMAN nanoarchaeal lineage. In several of the sunken biofilms,more » nanoarchaea comprise 10 to 25% of the community, based on fluorescent in situ hybridization and metagenomic analyses. Comparative community proteomic analyses show a persistence of bacterial proteins in sunken biofilms, but there is clear evidence for amino acid modifications due to acid hydrolysis. Given the low representation of bacterial cells in sunken biofilms based on microscopy, we infer that hydrolysis reflects proteins derived from lysed cells. For archaea, we detected 2,400 distinct proteins, including a subset involved in proteolysis and peptide uptake. Laboratory cultivation experiments using complex carbon substrates demonstrated anaerobic enrichment of Ferroplasma and Aplasma coupled to the reduction of ferric iron. These findings indicate dominance of acidophilic archaea in degrading biofilms and suggest that they play roles in anaerobic nutrient cycling at low pH.« less

  18. Impact of ZnO embedded feed spacer on biofilm development in membrane systems.

    PubMed

    Ronen, Avner; Semiat, Raphael; Dosoretz, Carlos G

    2013-11-01

    The concept of suppressing biofouling formation using an antibacterial feed spacer was investigated in a bench scale-cross flow system mimicking a spiral wound membrane configuration. An antibacterial composite spacer containing zinc oxide-nanoparticles was constructed by modification of a commercial polypropylene feed spacer using sonochemical deposition. The ability of the modified spacers to repress biofilm development on membranes was evaluated in flow-through cells simulating the flow conditions in commercial spiral wound modules. The experiments were performed at laminar flow (Re = 300) with a 200 kDa molecular weight cut off polysulfone ultrafiltration membrane using Pseudomonas putida S-12 as model biofilm bacteria. The modified spacers reduced permeate flux decrease at least by 50% compared to the unmodified spacers (control). The physical properties of the modified spacer and biofilm development were evaluated using high resolution/energy dispersive spectrometry-scanning electron microscopy, atomic force microscopy and confocal laser scanning microscopy imaging (HRSEM, EDS, AFM and CLSM). HRSEM images depicted significantly less bacteria attached to the membranes exposed to the modified spacer, mainly scattered and in a sporadic monolayer structure. AFM analysis indicated the influence of the modification on the spacer surface including a phase change on the upper surface. Dead-live staining assay by CLSM indicated that most of the bacterial cells attached on the membranes exposed to the modified spacer were dead in contrast to a developed biofilm which was predominant in the control samples. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. pH-activated Nanoparticles for Controlled Topical Delivery of Farnesol to Disrupt Oral Biofilm Virulence

    PubMed Central

    Horev, Benjamin; Klein, Marlise I.; Hwang, Geelsu; Li, Yong; Kim, Dongyeop; Koo, Hyun; Benoit, Danielle S.W.

    2015-01-01

    Development of effective therapies to control oral biofilms is challenging, as topically introduced agents must avoid rapid clearance from biofilm-tooth interfaces while targeting biofilm microenvironments. Additionally, exopolysaccharide matrix and acidification of biofilm microenvironments are associated with cariogenic (caries-producing) biofilm virulence. Thus, nanoparticle carriers capable of binding to hydroxyapatite (HA), saliva-coated HA (sHA), and exopolysaccharides with enhanced drug-release at acidic pH were developed. Nanoparticles are formed from diblock copolymers composed of 2-(dimethylamino)ethyl methacrylate (DMAEMA), butyl methacrylate (BMA), and 2-propylacrylic acid (PAA) (p(DMAEMA)-b-p(DMAEMA-co-BMA-co-PAA)) that self-assemble into ~21 nm cationic nanoparticles. Nanoparticles exhibit outstanding adsorption affinities (~244 L-mmol−1) to negatively-charged HA, sHA, and exopolysaccharide-coated sHA due to strong electrostatic interactions via multivalent tertiary amines of p(DMAEMA). Owing to hydrophobic cores, Nanoparticles load farnesol, a hydrophobic antibacterial drug, at ~22 wt%. Farnesol release is pH-dependent with t1/2=7 and 15 h for release at pH 4.5 and 7.2, as Nanoparticles undergo core destabilization at acidic pH, characteristic of cariogenic biofilm microenvironments. Importantly, topical applications of farnesol-loaded nanoparticles disrupted Streptococcus mutans biofilms 4-fold more effectively than free farnesol. Mechanical stability of biofilms treated with drug-loaded nanoparticles was compromised, resulting in >2-fold enhancement in biofilm removal under shear stress compared to free farnesol and controls. Farnesol-loaded nanoparticles effectively attenuated biofilm virulence in vivo using a clinically-relevant topical treatment regimen (2×/day) in a rodent dental caries disease model. Treatment with farnesol-loaded nanoparticles reduced both the number and severity of carious lesions, while free-farnesol had no effect

  20. Streptococcus mutans Extracellular DNA Is Upregulated during Growth in Biofilms, Actively Released via Membrane Vesicles, and Influenced by Components of the Protein Secretion Machinery

    PubMed Central

    Liao, Sumei; Klein, Marlise I.; Heim, Kyle P.; Fan, Yuwei; Bitoun, Jacob P.; Ahn, San-Joon; Burne, Robert A.; Koo, Hyun; Brady, L. Jeannine

    2014-01-01

    Streptococcus mutans, a major etiological agent of human dental caries, lives primarily on the tooth surface in biofilms. Limited information is available concerning the extracellular DNA (eDNA) as a scaffolding matrix in S. mutans biofilms. This study demonstrates that S. mutans produces eDNA by multiple avenues, including lysis-independent membrane vesicles. Unlike eDNAs from cell lysis that were abundant and mainly concentrated around broken cells or cell debris with floating open ends, eDNAs produced via the lysis-independent pathway appeared scattered but in a structured network under scanning electron microscopy. Compared to eDNA production of planktonic cultures, eDNA production in 5- and 24-h biofilms was increased by >3- and >1.6-fold, respectively. The addition of DNase I to growth medium significantly reduced biofilm formation. In an in vitro adherence assay, added chromosomal DNA alone had a limited effect on S. mutans adherence to saliva-coated hydroxylapatite beads, but in conjunction with glucans synthesized using purified glucosyltransferase B, the adherence was significantly enhanced. Deletion of sortase A, the transpeptidase that covalently couples multiple surface-associated proteins to the cell wall peptidoglycan, significantly reduced eDNA in both planktonic and biofilm cultures. Sortase A deficiency did not have a significant effect on membrane vesicle production; however, the protein profile of the mutant membrane vesicles was significantly altered, including reduction of adhesin P1 and glucan-binding proteins B and C. Relative to the wild type, deficiency of protein secretion and membrane protein insertion machinery components, including Ffh, YidC1, and YidC2, also caused significant reductions in eDNA. PMID:24748612

  1. Hollow fiber membrane based H₂ diffusion for efficient in situ biogas upgrading in an anaerobic reactor.

    PubMed

    Luo, Gang; Angelidaki, Irini

    2013-04-01

    Bubbleless gas transfer through a hollow fiber membrane (HFM) module was used to supply H2 to an anaerobic reactor for in situ biogas upgrading, and it creates a novel system that could achieve a CH4 content higher than 90 % in the biogas. The increase of CH4 content and pH, and the decrease of bicarbonate concentration were related with the increase of the H2 flow rate. The CH4 content increased from 78.4 % to 90.2 % with the increase of the H2 flow rate from 930 to 1,440 ml/(l  day), while the pH in the reactor remained below 8.0. An even higher CH4 content (96.1 %) was achieved when the H2 flow rate was increased to 1,760 ml/(l  day); however, the pH increased to around 8.3 due to bicarbonate consumption which hampered the anaerobic process. The biofilm formed on the HFM was found not to be beneficial for the process since it increased the resistance of H2 diffusion to the liquid. The study also demonstrated that the biofilm formed on the membrane only contributed 22-36 % to the H2 consumption, while most of the H2 was consumed by the microorganisms in the liquid phase.

  2. New functional biocarriers for enhancing the performance of a hybrid moving bed biofilm reactor-membrane bioreactor system.

    PubMed

    Deng, Lijuan; Guo, Wenshan; Ngo, Huu Hao; Zhang, Xinbo; Wang, Xiaochang C; Zhang, Qionghua; Chen, Rong

    2016-05-01

    In this study, new sponge modified plastic carriers for moving bed biofilm reactor (MBBR) was developed. The performance and membrane fouling behavior of a hybrid MBBR-membrane bioreactor (MBBR-MBR) system were also evaluated. Comparing to the MBBR with plastic carriers (MBBR), the MBBR with sponge modified biocarriers (S-MBBR) showed better effluent quality and enhanced nutrient removal at HRTs of 12h and 6h. Regarding fouling issue of the hybrid systems, soluble microbial products (SMP) of the MBR unit greatly influenced membrane fouling. The sponge modified biocarriers could lower the levels of SMP in mixed liquor and extracellular polymeric substances in activated sludge, thereby mitigating cake layer and pore blocking resistances of the membrane. The reduced SMP and biopolymer clusters in membrane cake layer were also observed. The results demonstrated that the sponge modified biocarriers were capable of improving overall MBBR performance and substantially alleviated membrane fouling of the subsequent MBR unit. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. GO-guided direct growth of highly oriented metal-organic framework nanosheet membranes for H2/CO2 separation.

    PubMed

    Li, Yujia; Liu, Haiou; Wang, Huanting; Qiu, Jieshan; Zhang, Xiongfu

    2018-05-07

    Highly oriented, ultrathin metal-organic framework (MOF) membranes are attractive for practical separation applications, but the scalable preparation of such membranes especially on standard tubular supports remains a huge challenge. Here we report a novel bottom-up strategy for directly growing a highly oriented Zn 2 (bIm) 4 (bIm = benzimidazole) ZIF nanosheet tubular membrane, based on graphene oxide (GO) guided self-conversion of ZnO nanoparticles (NPs). Through our approach, a thin layer of ZnO NPs confined between a substrate and a GO ultrathin layer self-converts into a highly oriented Zn 2 (bIm) 4 nanosheet membrane. The resulting membrane with a thickness of around 200 nm demonstrates excellent H 2 /CO 2 gas separation performance with a H 2 performance of 1.4 × 10 -7 mol m -2 s -1 Pa -1 and an ideal separation selectivity of about 106. The method can be easily scaled up and extended to the synthesis of other types of Zn-based MOF nanosheet membranes. Importantly, our strategy is particularly suitable for the large-scale fabrication of tubular MOF membranes that has not been possible through other methods.

  4. Utilization of membranes for H2O recycle system

    NASA Technical Reports Server (NTRS)

    Ohya, H.; Oguchi, M.

    1986-01-01

    Conceptual studies of closed ecological life support systems (CELSS) carried out at NAL in Japan for a water recycle system using membranes are reviewed. The system will treat water from shower room, urine, impure condensation from gas recycle system, and so on. The H2O recycle system is composed of prefilter, ultrafiltration membrane, reverse osmosis membrane, and distillator. Some results are shown for a bullet train of toilet-flushing water recycle equipment with an ultraviltration membrane module. The constant value of the permeation rate with a 4.7 square meters of module is about 70 1/h after 500th of operation. Thermovaporization with porous polytetrafluorocarbon membrane is also proposed to replce the distillator.

  5. Evaluation of the impact of organic material on the anaerobic methane and ammonium removal in a membrane aerated biofilm reactor (MABR) based on the multispecies biofilm modeling.

    PubMed

    Wu, Jun; Zhang, Yue

    2017-01-01

    The simultaneous nitrogen and methane removal by the combined nitritation, anaerobic ammonium oxidation (anammox), and nitrite dependent anaerobic methane oxidation (n-damo) processes in the membrane aerated biofilm reactor (MABR) offers clear advantages in term of energy saving and greenhouse gas emission mitigation. The rejected water from sludge digestion usually contained high ammonium, COD, and dissolved methane. The impact of influent COD on the anaerobic methane and ammonium removal in an MABR was evaluated in the model based study. The results indicated that the influent COD did not reduce the methane and ammonium removal efficiency at C/N ratio (influent COD/NH 4 + -N) less than 0.1. At high C/N ratio, the oxygen transfer coefficient needed to be increased to achieve high methane and nitrogen removal. Substrate flux analysis indicated that heterotrophic denitrification in the outside layer of biofilm reduced the impact of influent COD. Heterotrophic growth needed to be limited at the outside layer by using NO 3 - as electron acceptor; otherwise, the heterotrophic bacteria would compete NO 2 - and space with anammox and n-damo bacteria in the inner layers and reduce the nitrogen and methane removal efficiency.

  6. Time-kill kinetic analysis of antimicrobial chemotherapy based on hydrogen peroxide photolysis against Streptococcus mutans biofilm.

    PubMed

    Shirato, Midori; Nakamura, Keisuke; Kanno, Taro; Lingström, Peter; Niwano, Yoshimi; Örtengren, Ulf

    2017-08-01

    A recently developed antimicrobial technique utilizing hydroxyl radicals generated by hydrogen peroxide (H 2 O 2 ) photolysis represents a promising new therapy for preventing and treating dental caries. The present study compared the antimicrobial time-kill kinetics of H 2 O 2 photolysis, conventional antiseptics, and antimicrobial photodynamic therapy (aPDT) against biofilm-forming Streptococcus mutans (cariogenic bacteria) grown on hydroxyapatite disks. H 2 O 2 photolysis was performed by irradiating the biofilm immersed in 3% H 2 O 2 with 365-nm light-emitting diode (LED) light at an irradiance of 1000mW/cm 2 for up to 1.5min. Antiseptic treatments consisted of 0.2% chlorhexidine gluconate, 0.5% povidone-iodine, and 3% H 2 O 2 . The biofilm was immersed in each antiseptic for up to 4min. aPDT was performed by irradiating the biofilm immersed in 100μM methylene blue or toluidine blue O with 655-nm laser light at 1000mW/cm 2 for up to 4min. Based on the time-kill assay, the decimal reduction value (D-value) of each treatment was determined. With a D-value of 0.06min, H 2 O 2 photolysis exhibited the highest bactericidal effect against biofilm-forming S. mutans. In contrast, antiseptics and aPDT exerted a slower bactericidal effect, with D-values of 0.9-2.7min. In conclusion, the antimicrobial technique based on H 2 O 2 photolysis using 365-nm LED represents a strong adjunctive chemotherapy for dental caries treatment. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. A Flexible Optical pH Sensor Based on Polysulfone Membranes Coated with pH-Responsive Polyaniline Nanofibers

    PubMed Central

    Abu-Thabit, Nedal; Umar, Yunusa; Ratemi, Elaref; Ahmad, Ayman; Ahmad Abuilaiwi, Faraj

    2016-01-01

    A new optical pH sensor based on polysulfone (PSU) and polyaniline (PANI) was developed. A transparent and flexible PSU membrane was employed as a support. The electrically conductive and pH-responsive PANI was deposited onto the membrane surface by in situ chemical oxidative polymerization (COP). The absorption spectra of the PANI-coated PSU membranes exhibited sensitivity to pH changes in the range of 4–12, which allowed for designing a dual wavelength pH optical sensor. The performance of the membranes was assessed by measuring their response starting from high pH and going down to low pH, and vice versa. It was found that it is necessary to precondition the sensor layers before each measurement due to the slight hysteresis observed during forward and backward pH titrations. PSU membranes with polyaniline coating thicknesses in the range of ≈100–200 nm exhibited fast response times of <4 s, which are attributed to the porous, rough and nanofibrillar morphology of the polyaniline coating. The fabricated pH sensor was characterized by a sigmoidal response (R2 = 0.997) which allows for pH determination over a wide dynamic range. All membranes were stable for a period of more than six months when stored in 1 M HCl solution. The reproducibility of the fabricated optical pH sensors was found to be <0.02 absorption units after one month storage in 1 M HCl solution. The performance of the optical pH sensor was tested and the obtained pH values were compared with the results obtained using a pH meter device. PMID:27355953

  8. Embedded biofilm, a new biofilm model based on the embedded growth of bacteria.

    PubMed

    Jung, Yong-Gyun; Choi, Jungil; Kim, Soo-Kyoung; Lee, Joon-Hee; Kwon, Sunghoon

    2015-01-01

    A variety of systems have been developed to study biofilm formation. However, most systems are based on the surface-attached growth of microbes under shear stress. In this study, we designed a microfluidic channel device, called a microfluidic agarose channel (MAC), and found that microbial cells in the MAC system formed an embedded cell aggregative structure (ECAS). ECASs were generated from the embedded growth of bacterial cells in an agarose matrix and better mimicked the clinical environment of biofilms formed within mucus or host tissue under shear-free conditions. ECASs were developed with the production of extracellular polymeric substances (EPS), the most important feature of biofilms, and eventually burst to release planktonic cells, which resembles the full developmental cycle of biofilms. Chemical and genetic effects have also confirmed that ECASs are a type of biofilm. Unlike the conventional biofilms formed in the flow cell model system, this embedded-type biofilm completes the developmental cycle in only 9 to 12 h and can easily be observed with ordinary microscopes. We suggest that ECASs are a type of biofilm and that the MAC is a system for observing biofilm formation. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. The effect of pH and buffer concentration on anode biofilms of Thermincola ferriacetica.

    PubMed

    Lusk, Bradley G; Parameswaran, Prathap; Popat, Sudeep C; Rittmann, Bruce E; Torres, Cesar I

    2016-12-01

    We assessed the effects of pH and buffer concentration on current production and growth of biofilms of Thermincola ferriacetica - a thermophilic, Gram-positive, anode-respiring bacterium (ARB) - grown on anodes poised at a potential of -0.06V vs. SHE in microbial electrolysis cells (MECs) at 60°C. T. ferriacetica generated current in the pH range of 5.2 to 8.3 with acetate as the electron donor and 50mM bicarbonate buffer. Maximum current density was reduced by ~80% at pH5.2 and ~14% at 7.0 compared to pH8.3. Increasing bicarbonate buffer concentrations from 10mM to 100mM resulted in an increase in the current density by 40±6%, from 6.8±1.1 to 11.2±2.7Am(-2), supporting that more buffer alleviated pH depression within T. ferriacetica biofilms. Confocal laser scanning microscopy (CLSM) images indicated that higher bicarbonate buffer concentrations resulted in larger live biofilm thicknesses: from 68±20μm at 10mM bicarbonate to >150μm at 100mM, supporting that buffer availability was a strong influence on biofilm thickness. In comparison to mesophilic Geobacter sulfurreducens biofilms, the faster transport rates at higher temperature and the ability to grow at relatively lower pH allowed T. ferriacetica to produce higher current densities with lower buffer concentrations. Published by Elsevier B.V.

  10. Investigation of H2S separation from H2S/CH4 mixtures using functionalized and non-functionalized vertically aligned carbon nanotube membranes

    NASA Astrophysics Data System (ADS)

    Gilani, Neda; Towfighi, Jafar; Rashidi, Alimorad; Mohammadi, Toraj; Omidkhah, Mohammad Reza; Sadeghian, Ahmad

    2013-04-01

    Separation of H2S from binary mixtures of H2S/CH4 using vertically aligned carbon nanotube membranes fabricated in anodic aluminum oxide (AAO) template was studied experimentally. Carbon nanotubes (CNTs) were grown in five AAO templates with different pore diameters using chemical vapor deposition, and CNT/AAO membranes with tubular carbon nanotube structure and open caps were selected for separation of H2S. For this, two tubular CNT/AAO membranes were fabricated with the CNT inner diameters of 23 and 8 nm. It was found that permeability and selectivity of the membrane with inner diameter of 23 nm for CNT were independent of upstream feed pressure and H2S feed concentration unlike that of CNT having an inner diameter of 8 nm. Selectivity of these membranes for separation of H2S was obtained in the ranges of 1.36-1.58 and 2.11-2.86, for CNTs with internal diameters of 23 and 8 nm, respectively. In order to enhance the separation of H2S from H2S/CH4 mixtures, dodecylamine was used to functionalize the CNT/AAO membrane with higher selectivity. The results showed that for amido-functionalized membrane, both upstream feed pressure and H2S partial pressure in the feed significantly increased H2S permeability, and selectivity for H2S being in the range of 3.0-5.57 respectively.

  11. The CO 2 permeability and mixed gas CO 2/H 2 selectivity of membranes composed of CO 2-philic polymers

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Barillas, Mary Katharine; Enick, Robert M.; O’Brien, Michael

    2011-04-01

    The objective of this work was to design polymeric membranes that have very high CO 2 permeability and high mixed gas selectivity toward CO 2 rather than hydrogen. Therefore the membranes were based on "CO 2-philic" polymers that exhibit thermodynamically favorable Lewis acid:Lewis base and hydrogen bonding interactions with CO 2. CO 2-philic polymers that are solid at ambient temperature include polyfluoroacrylate (PFA); polyvinyl acetate (PVAc); and amorphous polylactic acid (PLA). Literature CO 2 permeability values for PVAc and PLA are disappointingly low. The cast PFA membranes from this study had low permeabilities (45 barrers at 25º C) and verymore » low CO 2/H 2 selectivity of 1.4. CO 2-philic polymers that are liquid at ambient conditions include polyethylene glycol (PEG), polypropylene glycol (PPG), polybutylene glycol with a linear -((CH 2) 4O)-repeat unit (i.e., polytetramethylene ether glycol (PTMEG)), polybutylene glycol (PBG) with a branched repeat unit, perfluoropolyether (PFPE), poly(dimethyl siloxane) (PDMS), and polyacetoxy oxetane (PAO). A small compound, glycerol triacetate (GTA) was also considered because it is similar in chemical structure to a trimer of PVAc. These liquids were tested as supported liquid membranes (SLM) and also (with the exception of PAD and GTA) as rubbery, crosslinked materials. Mixed gas permeability was measured using equimolar mixtures of CO 2 and H 2 feed streams at one atmosphere total pressure in steady-state flux experiments over the 298-423 K temperature range. The most promising SLMs were those composed of PEG, PTMEG, GTA, and PDMS. For example, at 37º C the PEG-, PTMEG-, GTA- and PDMS-based SLMs exhibited CO 2/H 2 selectivity values of ~11, 9, 9, and 3.5, respectively, and CO 2 permeability values of ~800, 900, 1900, and 2000 barrers, respectively. Crosslinked versions of the PEG, PTMEG and PDMS membranes at 37º C exhibited selectivity values of ~5, 6, and 3.5, respectively, and CO 2 permeability values of

  12. Biofilm growth on polyvinylchloride surface incubated in suboptimal microbial warm water and effect of sanitizers on biofilm removal post biofilm formation.

    PubMed

    Maharjan, Pramir; Huff, Geraldine; Zhang, Wen; Watkins, Susan

    2017-01-01

    An in vitro experiment was conducted to understand the nature of biofilm growth on polyvinyl chloride (PVC) surface when exposed to suboptimal-quality microbial water (>4 log 10 cfu/mL) obtained from a poultry drinking water source mimicking water in waterlines during the first week of poultry brooding condition. PVC sections (internal surface area of 15.16 cm 2 ) were utilized in the study to grow biofilm. After a 7-d test period, test coupons with 7-day-old biofilm were transferred into autoclaved municipal water and then treated with either chlorine-based or hydrogen peroxide-based sanitizer at bird drinking water rate, to see the impact on removal of biofilm formed on test coupons. Two trials (T1 and T2) were conducted. Test coupons used in T1 and T2 had the bacterial growth of 3.67 (SEM 0.04) and 3.97 (SEM 0.11) log 10 cfu/cm 2 on d 7. After sanitizer application, chlorine-based sanitizer removed bacteria in biofilm completely (0 cfu/cm 2 ) within 24 h post treatment whereas hydrogen peroxide-based sanitizer reduced the counts to 1.68 log 10 cfu/cm 2 (P < 0.05) by 48 h post sanitizer application. Control remained the same (P > 0.05). Results indicated that biofilm formation can occur quickly under suboptimal water condition on PVC surface, and sanitizer application helped mitigate already formed biofilm, yet chlorine proved to be more effective than hydrogen peroxide. © 2016 Poultry Science Association Inc.

  13. Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes in biofilms by pulsed ultraviolet light.

    PubMed

    Montgomery, Nedra L; Banerjee, Pratik

    2015-06-10

    The inactivation of biofilms formed by pathogenic bacteria on ready-to-eat and minimally processed fruits and vegetables by nonthermal processing methods is critical to ensure food safety. Pulsed ultraviolet (PUV) light has shown promise in the surface decontamination of liquid, powdered, and solid foods. In this study, the antimicrobial efficacy of PUV light treatment on nascent biofilms formed by Escherichia coli O157:H7 and Listeria monocytogenes on the surfaces of food packaging materials, such as low-density polyethylene (LDPE), and fresh produce, such as lettuce (Lactuca sativa) leaves, was investigated. The formation of biofilms on Romaine lettuce leaves and LDPE films was confirmed by crystal violet and Alcian blue staining methods. Inactivation of cells in the biofilm was determined by standard plating procedures, and by a luminescence-based bacterial cell viability assay. Upon PUV treatment of 10 s at two different light source to sample distances (4.5 and 8.8 cm), viable cell counts of L. monocytogenes and E. coli O157:H7 in biofilms on the lettuce surface were reduced by 0.6-2.2 log CFU mL(-1) and 1.1-3.8 log CFU mL(-1), respectively. On the LDPE surface, the efficiency of inactivation of biofilm-encased cells was slightly higher. The maximum values for microbial reduction on LDPE were 2.7 log CFU mL(-1) and 3.9 log CFU mL(-1) for L. monocytogenes and E. coli O157:H7, respectively. Increasing the duration of PUV light exposure resulted in a significant (P < 0.05) reduction in biofilm formation by both organisms. The results also revealed that PUV treatment was more effective at reducing E. coli biofilms compared with Listeria biofilms. A moderate increase in temperature (~7-15°C) was observed for both test materials. PUV is an effective nonthermal intervention method for surface decontamination of E. coli O157:H7 and L. monocytogenes on fresh produce and packaging materials.

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

    PubMed

    Schlafer, Sebastian; Raarup, Merete K; Meyer, Rikke L; Sutherland, Duncan S; Dige, Irene; Nyengaard, Jens R; Nyvad, Bente

    2011-01-01

    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. 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. The designed biofilm model represents a promising tool to determine the effect of potential therapeutic agents on biofilm growth, composition and extracellular pH. Ratiometric pH analysis using C-SNARF-4 gives detailed

  15. Evaluation of zosteric acid for mitigating biofilm formation of Pseudomonas putida isolated from a membrane bioreactor system.

    PubMed

    Polo, Andrea; Foladori, Paola; Ponti, Benedetta; Bettinetti, Roberta; Gambino, Michela; Villa, Federica; Cappitelli, Francesca

    2014-05-28

    This study provides data to define an efficient biocide-free strategy based on zosteric acid to counteract biofilm formation on the membranes of submerged bioreactor system plants. 16S rRNA gene phylogenetic analysis showed that gammaproteobacteria was the prevalent taxa on fouled membranes of an Italian wastewater plant. Pseudomonas was the prevalent genus among the cultivable membrane-fouler bacteria and Pseudomonas putida was selected as the target microorganism to test the efficacy of the antifoulant. Zosteric acid was not a source of carbon and energy for P. putida cells and, at 200 mg/L, it caused a reduction of bacterial coverage by 80%. Biofilm experiments confirmed the compound caused a significant decrease in biomass (-97%) and thickness (-50%), and it induced a migration activity of the peritrichous flagellated P. putida over the polycarbonate surface not amenable to a biofilm phenotype. The low octanol-water partitioning coefficient and the high water solubility suggested a low bioaccumulation potential and the water compartment as its main environmental recipient and capacitor. Preliminary ecotoxicological tests did not highlight direct toxicity effects toward Daphnia magna. For green algae Pseudokirchneriella subcapitata an effect was observed at concentrations above 100 mg/L with a significant growth of protozoa that may be connected to a concurrent algal growth inhibition.

  16. Evaluation of Zosteric Acid for Mitigating Biofilm Formation of Pseudomonas putida Isolated from a Membrane Bioreactor System

    PubMed Central

    Polo, Andrea; Foladori, Paola; Ponti, Benedetta; Bettinetti, Roberta; Gambino, Michela; Villa, Federica; Cappitelli, Francesca

    2014-01-01

    This study provides data to define an efficient biocide-free strategy based on zosteric acid to counteract biofilm formation on the membranes of submerged bioreactor system plants. 16S rRNA gene phylogenetic analysis showed that gammaproteobacteria was the prevalent taxa on fouled membranes of an Italian wastewater plant. Pseudomonas was the prevalent genus among the cultivable membrane-fouler bacteria and Pseudomonas putida was selected as the target microorganism to test the efficacy of the antifoulant. Zosteric acid was not a source of carbon and energy for P. putida cells and, at 200 mg/L, it caused a reduction of bacterial coverage by 80%. Biofilm experiments confirmed the compound caused a significant decrease in biomass (−97%) and thickness (−50%), and it induced a migration activity of the peritrichous flagellated P. putida over the polycarbonate surface not amenable to a biofilm phenotype. The low octanol-water partitioning coefficient and the high water solubility suggested a low bioaccumulation potential and the water compartment as its main environmental recipient and capacitor. Preliminary ecotoxicological tests did not highlight direct toxicity effects toward Daphnia magna. For green algae Pseudokirchneriella subcapitata an effect was observed at concentrations above 100 mg/L with a significant growth of protozoa that may be connected to a concurrent algal growth inhibition. PMID:24879523

  17. Chemorepulsion from the Quorum Signal Autoinducer-2 Promotes Helicobacter pylori Biofilm Dispersal

    PubMed Central

    Anderson, Jeneva K.; Huang, Julie Y.; Wreden, Christopher; Sweeney, Emily Goers; Goers, John; Remington, S. James

    2015-01-01

    ABSTRACT The gastric pathogen Helicobacter pylori forms biofilms on abiotic and biotic surfaces. We have shown previously that H. pylori perceives the quorum signal autoinducer-2 (AI-2) as a chemorepellent. We report here that H. pylori chemorepulsion from endogenous AI-2 influences the proportions and spatial organization of cells within biofilms. Strains that fail to produce AI-2 (∆luxS strains) or are defective for chemotaxis (∆cheA strains) formed more spatially homogenous biofilms with a greater proportion of adherent versus planktonic cells than wild-type biofilms. Reciprocally, a strain that overproduced AI-2 (luxSOP) formed biofilms with proportionally fewer adherent cells. Along with the known AI-2 chemoreceptor, TlpB, we identified AibA and AibB, two novel periplasmic binding proteins that are required for the AI-2 chemorepulsion response. Disruptions in any of the proteins required for AI-2 chemotaxis recapitulated the biofilm adherence and spatial organization phenotype of the ∆luxS mutant. Furthermore, exogenous administration of AI-2 was sufficient to decrease the proportion of adherent cells in biofilms and promote dispersal of cells from biofilms in a chemotaxis-dependent manner. Finally, we found that disruption of AI-2 production or AI-2 chemotaxis resulted in increased clustering of cells in microcolonies on cultured epithelial cells. We conclude that chemotaxis from AI-2 is a determinant of H. pylori biofilm spatial organization and dispersal. PMID:26152582

  18. Outer membrane lipoprotein VacJ is required for the membrane integrity, serum resistance and biofilm formation of Actinobacillus pleuropneumoniae.

    PubMed

    Xie, Fang; Li, Gang; Zhang, Wanjiang; Zhang, Yanhe; Zhou, Long; Liu, Shuanghong; Liu, Siguo; Wang, Chunlai

    2016-02-01

    The outer membrane proteins of Actinobacillus pleuropneumoniae are mediators of infection, acting as targets for the host's defense system. The outer membrane lipoprotein VacJ is involved in serum resistance and intercellular spreading in several pathogenic bacteria. To investigate the role of VacJ in the pathogenicity of Actinobacillus pleuropneumoniae, the vacJ gene-deletion mutant MD12 ΔvacJ was constructed. The increased susceptibility to KCl, SDS plus EDTA, and several antibiotics in the MD12ΔvacJ mutant suggested that the stability of the outer membrane was impaired as a result of the mutation in the vacJ gene. The increased NPN fluorescence and significant cellular morphological variation in the MD12ΔvacJ mutant further demonstrated the crucial role of the VacJ lipoprotein in maintaining the outer membrane integrity of A. pleuropneumoniae. In addition, the MD12ΔvacJ mutant exhibited decreased survival from the serum and complement killing compared to the wild-type strain. Interestingly, the MD12ΔvacJ mutant showed reduced biofilm formation compared to the wild-type strain. To our knowledge, this is the first description of the VacJ lipoprotein contributing to bacterial biofilm formation. The data presented in this study illustrate the important role of the VacJ lipoprotein in the maintenance of cellular integrity, serum resistance, and biofilm formation in A. pleuropneumoniae. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Biofilm Formation on Reverse Osmosis Membranes Is Initiated and Dominated by Sphingomonas spp.▿ †

    PubMed Central

    Bereschenko, L. A.; Stams, A. J. M.; Euverink, G. J. W.; van Loosdrecht, M. C. M.

    2010-01-01

    The initial formation and spatiotemporal development of microbial biofilm layers on surfaces of new and clean reverse osmosis (RO) membranes and feed-side spacers were monitored in situ using flow cells placed in parallel with the RO system of a full-scale water treatment plant. The feed water of the RO system had been treated by the sequential application of coagulation, flocculation, sand filtration, ultrafiltration, and cartridge filtration processes. The design of the flow cells permitted the production of permeate under cross-flow conditions similar to those in spiral-wound RO membrane elements of the full-scale system. Membrane autopsies were done after 4, 8, 16, and 32 days of flow-cell operation. A combination of molecular (fluorescence in situ hybridization [FISH], denaturing gradient gel electrophoresis [DGGE], and cloning) and microscopic (field emission scanning electron, epifluorescence, and confocal laser scanning microscopy) techniques was applied to analyze the abundance, composition, architecture, and three-dimensional structure of biofilm communities. The results of the study point out the unique role of Sphingomonas spp. in the initial formation and subsequent maturation of biofilms on the RO membrane and feed-side spacer surfaces. PMID:20190090

  20. Lactobacillus brevis CD2 inhibits Prevotella melaninogenica biofilm.

    PubMed

    Vuotto, C; Barbanti, F; Mastrantonio, P; Donelli, G

    2014-10-01

    To evaluate the ability of the probiotic strain Lactobacillus brevis CD2 to inhibit the opportunistic anaerobe Prevotella melaninogenica (PM1), a well-known causative agent of periodontitis. The inhibitory effect of Lactobacillus CD2 on Prevotella PM1 biofilm was assessed both by exposing the anaerobe to the supernatant of the probiotic strain and by growing the two strains to obtain single or mixed biofilms. The inhibitory effect of CD2 on PM1 was also checked by the agar overlay method. The development of PM1 biofilm was strongly affected (56% decrease in OD value) by the CD2 supernatant after 96 h. A dose-dependent biofilm reduction was also observed at 1/10 and 1/100 dilutions of supernatant. Confocal microscopy on the mixed biofilms revealed the ability of CD2 to prevail on PM1, greatly reducing the biofilm of the latter. It has been hypothesized a multifactorial nature of the inhibition mechanism, the strong adherence ability of CD2 strain together with the released metabolites presumably contributing to the reduction in the PM1 biofilm detected by confocal microscopy. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  1. Analysis of Microbial Communities in Biofilms from CSTR-Type Hollow Fiber Membrane Biofilm Reactors for Autotrophic Nitrification and Hydrogenotrophic Denitrification.

    PubMed

    Shin, Jung-Hun; Kim, Byung-Chun; Choi, Okkyoung; Kim, Hyunook; Sang, Byoung-In

    2015-10-01

    Two hollow fiber membrane biofilm reactors (HF-MBfRs) were operated for autotrophic nitrification and hydrogenotrophic denitrification for over 300 days. Oxygen and hydrogen were supplied through the hollow fiber membrane for nitrification and denitrification, respectively. During the period, the nitrogen was removed with the efficiency of 82-97% for ammonium and 87-97% for nitrate and with the nitrogen removal load of 0.09-0.26 kg NH4(+)-N/m(3)/d and 0.10-0.21 kg NO3(-)-N/m(3)/d, depending on hydraulic retention time variation by the two HF-MBfRs for autotrophic nitrification and hydrogenotrophic denitrification, respectively. Biofilms were collected from diverse topological positions in the reactors, each at different nitrogen loading rates, and the microbial communities were analyzed with partial 16S rRNA gene sequences in denaturing gradient gel electrophoresis (DGGE). Detected DGGE band sequences in the reactors were correlated with nitrification or denitrification. The profile of the DGGE bands depended on the NH4(+) or NO3(-) loading rate, but it was hard to find a major strain affecting the nitrogen removal efficiency. Nitrospira-related phylum was detected in all biofilm samples from the nitrification reactors. Paracoccus sp. and Aquaspirillum sp., which are an autohydrogenotrophic bacterium and an oligotrophic denitrifier, respectively, were observed in the denitrification reactors. The distribution of microbial communities was relatively stable at different nitrogen loading rates, and DGGE analysis based on 16S rRNA (341f /534r) could successfully detect nitrate-oxidizing and hydrogen-oxidizing bacteria but not ammonium-oxidizing bacteria in the HF-MBfRs.

  2. Kinetics of nitrate and perchlorate reduction in ion exchange brine using the membrane biofilm reactor (MBfR)

    EPA Science Inventory

    Several sources of bacterial inocula were tested for their ability to reduce nitrate and perchlorate in synthetic ion-exchange spent brine (3-4.5% salinity) using a hydrogen-based membrane biofilm reactor (MBfR). Nitrate and perchlorate removal fluxes reached as high as 5.4 g N ...

  3. Inactivation of Escherichia coli O157:H7 on stainless steel upon exposure to Paenibacillus polymyxa biofilms.

    PubMed

    Kim, Seonhwa; Bang, Jihyun; Kim, Hoikyung; Beuchat, Larry R; Ryu, Jee-Hoon

    2013-11-01

    We investigated the potential use of biofilm formed by a competitive-exclusion (CE) microorganism to inactivate Escherichia coli O157:H7 on a stainless steel surface. Five microorganisms showing inhibitory activities against E. coli O157:H7 were isolated from vegetable seeds and sprouts. The microorganism with the greatest antimicrobial activity was identified as Paenibacillus polymyxa (strain T5). In tryptic soy broth (TSB), strain T5 reached a higher population at 25 °C than at 12 or 37 °C without losing inhibitory activity against E. coli O157:H7. When P. polymyxa (6 log CFU/mL) was co-cultured with E. coli O157:H7 (2, 3, 4, or 5 log CFU/mL) in TSB at 25 °C, the number of E. coli O157:H7 decreased significantly within 24h. P. polymyxa formed a biofilm on stainless steel coupons (SSCs) in TSB at 25 °C within 24h, and cells in biofilms, compared to attached cells without biofilm formation, showed significantly increased resistance to a dry environment (43% relative humidity [RH]). With the exception of an inoculum of 4 log CFU/coupon at 100% RH, upon exposure to biofilm formed by P. polymyxa on SSCs, populations of E. coli O157:H7 (2, 4, or 6 log CFU/coupon) were significantly reduced within 48 h. Most notably, when E. coli O157:H7 at 2 log CFU/coupon was applied to SSCs on which P. polymyxa biofilm had formed, it was inactivated within 1h, regardless of RH. These results will be useful when developing strategies using biofilms produced by competitive exclusion microorganisms to inactivate foodborne pathogens in food processing environments. © 2013.

  4. Fabrication of COF-MOF Composite Membranes and Their Highly Selective Separation of H2/CO2.

    PubMed

    Fu, Jingru; Das, Saikat; Xing, Guolong; Ben, Teng; Valtchev, Valentin; Qiu, Shilun

    2016-06-22

    The search for new types of membrane materials has been of continuous interest in both academia and industry, given their importance in a plethora of applications, particularly for energy-efficient separation technology. In this contribution, we demonstrate for the first time that a metal-organic framework (MOF) can be grown on the covalent-organic framework (COF) membrane to fabricate COF-MOF composite membranes. The resultant COF-MOF composite membranes demonstrate higher separation selectivity of H2/CO2 gas mixtures than the individual COF and MOF membranes. A sound proof for the synergy between two porous materials is the fact that the COF-MOF composite membranes surpass the Robeson upper bound of polymer membranes for mixture separation of a H2/CO2 gas pair and are among the best gas separation MOF membranes reported thus far.

  5. Plant Biofilm Inhibitors to Discover Biofilm Genes

    DTIC Science & Technology

    2011-04-08

    REPORT Final Report for Plant Biofilm Inhibitors to Discover Biofilm Genes 14. ABSTRACT 16. SECURITY CLASSIFICATION OF: To control biofilms , we have...synthesized the natural biofilm inhibitor (5Z)-4-bromo-5-(bromomethylene) -3-butyl-2(5H)-furanone from the red alga Delisea pulchra and determined that...Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 15. SUBJECT TERMS biofilms , biofilm inhibitors Thomas K. Wood Texas Engineering

  6. Pb and Cd binding to natural freshwater biofilms developed at different pH: the important role of culture pH.

    PubMed

    Hua, Xiuyi; Dong, Deming; Ding, Xiaoou; Yang, Fan; Jiang, Xu; Guo, Zhiyong

    2013-01-01

    The effects of solution pH on adsorption of trace metals to different types of natural aquatic solid materials have been studied extensively, but few studies have been carried out to investigate the effect of pH at which the solid materials were formed on the adsorption. The purpose of present study is to examine this effect of culture pH on metal adsorption to natural freshwater biofilms. The adsorption of Pb and Cd to biofilms which were developed at different culture pH values (ranging from 6.5 to 9.0) was measured at the same adsorption pH value (6.5). The culture pH had considerable effects on both composition and metal adsorption ability of the biofilms. Higher culture pH usually promoted the accumulation of organic material and Fe oxides in the biofilms. The culture pH also affected the quantity and species of algae in the biofilms. The adsorption of Pb and Cd to the biofilms generally increased with the increase of culture pH. This increase was minor at lower pH range and significant at higher pH range and was more remarkable for Cd adsorption than for Pb adsorption. The notable contribution of organic material to the adsorption at higher culture pH values was also observed. The profound impacts of culture pH on adsorption behavior of biofilms mainly resulted from the variation of total contents of the biofilm components and were also affected by the alteration of composition and properties of the components.

  7. Anti-biofilm action of nitric oxide-releasing alkyl-modified poly(amidoamine) dendrimers against Streptococcus mutans.

    PubMed

    Backlund, Christopher J; Worley, Brittany V; Schoenfisch, Mark H

    2016-01-01

    The effect of nitric oxide (NO)-releasing dendrimer hydrophobicity on Streptococcus mutans killing and biofilm disruption was examined at pH 7.4 and 6.4, the latter relevant to dental caries. Generation 1 (G1) poly(amidoamine) (PAMAM) dendrimers were modified with alkyl epoxides to generate propyl-, butyl-, hexyl-, octyl-, and dodecyl-functionalized dendrimers. The resulting secondary amines were reacted with NO to form N-diazeniumdiolate NO donor-modified dendrimer scaffolds (total NO ∼1μmol/mg). The bactericidal action of the NO-releasing dendrimers against both planktonic and biofilm-based S. mutans proved greatest with increasing alkyl chain length and at lower pH. Improved bactericidal efficacy at pH 6.4 was attributed to increased scaffold surface charge that enhanced dendrimer-bacteria association and ensuing membrane damage. For shorter alkyl chain (i.e., propyl and butyl) dendrimer modifications, increased antibacterial action at pH 6.4 was due to faster NO-release kinetics from proton-labile N-diazeniumdiolate NO donors. Octyl- and dodecyl-modified PAMAM dendrimers proved most effective for eradicating S. mutans biofilms with NO release mitigating dendrimer scaffold cytotoxicity. We report the antibacterial and anti-biofilm efficacy of dual-action nitric oxide (NO)-releasing dendrimers against S. mutans, an etiological agent in dental caries. This work was undertaken to enhance the anti-biofilm action of these scaffolds by employing various alkyl chain modifications. Furthermore, we evaluated the ability of NO to eradicate cariogenic biofilms. We found that at the lower pH associated with dental caries (pH ∼6.4), NO has a more pronounced antibacterial effect for alkyl modifications less capable of biofilm penetration and membrane disruption. Of greatest significance, we introduce dendrimers as a new macromolecular antibacterial agent against the cariogenic bacteria S. mutans. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All

  8. Pyrosequencing analysis of microbial communities in hollow fiber-membrane biofilm reactors system for treating high-strength nitrogen wastewater.

    PubMed

    Park, Jung-Hun; Choi, Okkyoung; Lee, Tae-Ho; Kim, Hyunook; Sang, Byoung-In

    2016-11-01

    Wastewaters from swine farms, nitrogen-dealing industries or side-stream processes of a wastewater treatment plant (e.g., anaerobic digesters, sludge thickening processes, etc.) are characterized by low C/N ratios and not easily treatable. In this study, a hollow fiber-membrane biofilm reactors (HF-MBfR) system consisting of an O2-based HF-MBfR and an H2-based HF-MBfR was applied for treating high-strength wastewater. The reactors were continuously operated with low supply of O2 and H2 and without any supply of organic carbon for 250 d. Gradual increase of ammonium and nitrate concentration in the influent showed stable and high nitrogen removal efficiency, and the maximum ammonium and nitrate removal rates were 0.48 kg NH4(+)-N m(-3) d(-1) and 0.55 kg NO3(-)-N m(-3) d(-1), respectively. The analysis of the microbial communities using pyrosequencing analysis indicated that Nitrosospira multiformis, ammonium-oxidizing bacteria, and Nitrobacter winogradskyi and Nitrobacter vulgaris, nitrite-oxidizing bacteria were highly enriched in the O2-based HF-MBfR. In the H2-based HF-MBfR, hydrogenotrophic denitrifying bacteria belonging to the family of Thiobacillus and Comamonadaceae were initially dominant, but were replaced to heterotrophic denitrifiers belonging to Rhodocyclaceae and Rhodobacteraceae utilizing by-products induced from autotrophic denitrifying bacteria. The pyrosequencing analysis of microbial communities indicates that the autotrophic HF-MBfRs system well developed autotrophic nitrifying and denitrifying bacteria within a relatively short period to accomplish almost complete nitrogen removal. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Streptococcus oligofermentans Inhibits Streptococcus mutans in Biofilms at Both Neutral pH and Cariogenic Conditions.

    PubMed

    Bao, Xudong; de Soet, Johannes Jacob; Tong, Huichun; Gao, Xuejun; He, Libang; van Loveren, Cor; Deng, Dong Mei

    2015-01-01

    Homeostasis of oral microbiota can be maintained through microbial interactions. Previous studies showed that Streptococcus oligofermentans, a non-mutans streptococci frequently isolated from caries-free subjects, inhibited the cariogenic Streptococcus mutans by the production of hydrogen peroxide (HP). Since pH is a critical factor in caries formation, we aimed to study the influence of pH on the competition between S. oligofermentans and S. mutans in biofilms. To this end, S. mutans and S. oligofermentans were inoculated alone or mixed at 1:1 ratio in buffered biofilm medium in a 96-well active attachment model. The single- and dual-species biofilms were grown under either constantly neutral pH or pH-cycling conditions. The latter includes two cycles of 8 h neutral pH and 16 h pH 5.5, used to mimic cariogenic condition. The 48 h biofilms were analysed for the viable cell counts, lactate and HP production. The last two measurements were carried out after incubating the 48 h biofilms in buffers supplemented with 1% glucose (pH 7.0) for 4 h. The results showed that S. oligofermentans inhibited the growth of S. mutans in dual-species biofilms under both tested pH conditions. The lactic acid production of dual-species biofilms was significantly lower than that of single-species S. mutans biofilms. Moreover, dual-species and single-species S. oligofermentans biofilms grown under pH-cycling conditions (with a 16 h low pH period) produced a significantly higher amount of HP than those grown under constantly neutral pH. In conclusion, S. oligofermentans inhibited S. mutans in biofilms not only under neutral pH, but also under pH-cycling conditions, likely through HP production. S. oligofermentans may be a compelling probiotic candidate against caries.

  10. Towards Biofilm Spectroscopy - A Novel Microfluidic Approach for Characterizing Biofilm Subpopulation by Microwave-Based Electrical Impedance Spectroscopy

    NASA Astrophysics Data System (ADS)

    Richter, Christiane; Schneider, Stefan; Rapp, Bastian E.; Schmidt, Sönke; Schüßler, Martin; Jakoby, Rolf; Bruchmann, Julia; Bischer, Moritz; Schwartz, Thomas

    2018-03-01

    In this work three disciplines - microfluidics, microbiology and microwave engineering - are utilized to develop a system for analyzing subpopulations of biofilms and their reaction to antibiotic treatment. We present handling strategies to destabilize a biofilm inside a microfluidic system down to aggregate sizes of<10 µm2 as well as microfluidic structures for the flow-through filtration of the resulting cell suspensions. For the analysis of the cell populations by microwave electrical impedance spectroscopy, two novel calibration schemes are demonstrated to cover both, reflection as well as transmission measurements of dielectric fluids. The broadband calibration strategies are solely based on liquid standards and allow a precise long-term monitoring with a resolution up to Δ ɛ = 6 Δ = 1.5 ‰ at H 5 GHz. Combining these three research topics therefore will open up new ways for analyzing biofilm effects.

  11. Apple flavonoid phloretin inhibits Escherichia coli O157:H7 biofilm formation and ameliorates colon inflammation in rats.

    PubMed

    Lee, Jin-Hyung; Regmi, Sushil Chandra; Kim, Jung-Ae; Cho, Moo Hwan; Yun, Hyungdon; Lee, Chang-Soo; Lee, Jintae

    2011-12-01

    Pathogenic biofilms have been associated with persistent infections due to their high resistance to antimicrobial agents, while commensal biofilms often fortify the host's immune system. Hence, controlling biofilm formation of both pathogenic bacteria and commensal bacteria is important in bacterium-related diseases. We investigated the effect of plant flavonoids on biofilm formation of enterohemorrhagic Escherichia coli O157:H7. The antioxidant phloretin, which is abundant in apples, markedly reduced E. coli O157:H7 biofilm formation without affecting the growth of planktonic cells, while phloretin did not harm commensal E. coli K-12 biofilms. Also, phloretin reduced E. coli O157:H7 attachment to human colon epithelial cells. Global transcriptome analyses revealed that phloretin repressed toxin genes (hlyE and stx(2)), autoinducer-2 importer genes (lsrACDBF), curli genes (csgA and csgB), and dozens of prophage genes in E. coli O157:H7 biofilm cells. Electron microscopy confirmed that phloretin reduced fimbria production in E. coli O157:H7. Also, phloretin suppressed the tumor necrosis factor alpha-induced inflammatory response in vitro using human colonic epithelial cells. Moreover, in the rat model of colitis induced by trinitrobenzene sulfonic acid (TNBS), phloretin significantly ameliorated colon inflammation and body weight loss. Taken together, our results suggest that the antioxidant phloretin also acts as an inhibitor of E. coli O157:H7 biofilm formation as well as an anti-inflammatory agent in inflammatory bowel diseases without harming beneficial commensal E. coli biofilms.

  12. A proteomic investigation of Fusobacterium nucleatum alkaline-induced biofilms

    PubMed Central

    2012-01-01

    Background The Gram negative anaerobe Fusobacterium nucleatum has been implicated in the aetiology of periodontal diseases. Although frequently isolated from healthy dental plaque, its numbers and proportion increase in plaque associated with disease. One of the significant physico-chemical changes in the diseased gingival sulcus is increased environmental pH. When grown under controlled conditions in our laboratory, F. nucleatum subspecies polymorphum formed mono-culture biofilms when cultured at pH 8.2. Biofilm formation is a survival strategy for bacteria, often associated with altered physiology and increased virulence. A proteomic approach was used to understand the phenotypic changes in F. nucleatum cells associated with alkaline induced biofilms. The proteomic based identification of significantly altered proteins was verified where possible using additional methods including quantitative real-time PCR (qRT-PCR), enzyme assay, acidic end-product analysis, intracellular polyglucose assay and Western blotting. Results Of 421 proteins detected on two-dimensional electrophoresis gels, spot densities of 54 proteins varied significantly (p < 0.05) in F. nucleatum cultured at pH 8.2 compared to growth at pH 7.4. Proteins that were differentially produced in biofilm cells were associated with the functional classes; metabolic enzymes, transport, stress response and hypothetical proteins. Our results suggest that biofilm cells were more metabolically efficient than planktonic cells as changes to amino acid and glucose metabolism generated additional energy needed for survival in a sub-optimal environment. The intracellular concentration of stress response proteins including heat shock protein GroEL and recombinational protein RecA increased markedly in the alkaline environment. A significant finding was the increased abundance of an adhesin, Fusobacterial outer membrane protein A (FomA). This surface protein is known for its capacity to bind to a vast number of

  13. Explosive cell lysis as a mechanism for the biogenesis of bacterial membrane vesicles and biofilms

    PubMed Central

    Turnbull, Lynne; Toyofuku, Masanori; Hynen, Amelia L.; Kurosawa, Masaharu; Pessi, Gabriella; Petty, Nicola K.; Osvath, Sarah R.; Cárcamo-Oyarce, Gerardo; Gloag, Erin S.; Shimoni, Raz; Omasits, Ulrich; Ito, Satoshi; Yap, Xinhui; Monahan, Leigh G.; Cavaliere, Rosalia; Ahrens, Christian H.; Charles, Ian G.; Nomura, Nobuhiko; Eberl, Leo; Whitchurch, Cynthia B.

    2016-01-01

    Many bacteria produce extracellular and surface-associated components such as membrane vesicles (MVs), extracellular DNA and moonlighting cytosolic proteins for which the biogenesis and export pathways are not fully understood. Here we show that the explosive cell lysis of a sub-population of cells accounts for the liberation of cytosolic content in Pseudomonas aeruginosa biofilms. Super-resolution microscopy reveals that explosive cell lysis also produces shattered membrane fragments that rapidly form MVs. A prophage endolysin encoded within the R- and F-pyocin gene cluster is essential for explosive cell lysis. Endolysin-deficient mutants are defective in MV production and biofilm development, consistent with a crucial role in the biogenesis of MVs and liberation of extracellular DNA and other biofilm matrix components. Our findings reveal that explosive cell lysis, mediated through the activity of a cryptic prophage endolysin, acts as a mechanism for the production of bacterial MVs. PMID:27075392

  14. Antifungal activity, mode of action and anti-biofilm effects of Laurus nobilis Linnaeus essential oil against Candida spp.

    PubMed

    Peixoto, Larissa Rangel; Rosalen, Pedro Luiz; Ferreira, Gabriela Lacet Silva; Freires, Irlan Almeida; de Carvalho, Fabíola Galbiatti; Castellano, Lúcio Roberto; de Castro, Ricardo Dias

    2017-01-01

    The present study demonstrated the antifungal potential of the chemically characterized essential oil (EO) of Laurus nobilis L. (bay laurel) against Candida spp. biofilm adhesion and formation, and further established its mode of action on C. albicans. L. nobilis EO was obtained and tested for its minimum inhibitory and fungicidal concentrations (MIC/MFC) against Candida spp., as well as for interaction with cell wall biosynthesis and membrane ionic permeability. Then we evaluated its effects on the adhesion, formation, and reduction of 48hC. albicans biofilms. The EO phytochemical profile was determined by gas chromatography coupled to mass spectrometry (GC/MS). The MIC and MFC values of the EO ranged from (250 to 500) μg/mL. The MIC values increased in the presence of sorbitol (osmotic protector) and ergosterol, which indicates that the EO may affect cell wall biosynthesis and membrane ionic permeability, respectively. At 2 MIC the EO disrupted initial adhesion of C. albicans biofilms (p<0.05) and affected biofilm formation with no difference compared to nystatin (p>0.05). When applied for 1min, every 8h, for 24h and 48h, the EO reduced the amount of C. albicans mature biofilm with no difference in relation to nystatin (p>0.05). The phytochemical analysis identified isoeugenol as the major compound (53.49%) in the sample. L. nobilis EO has antifungal activity probably due to monoterpenes and sesquiterpenes in its composition. This EO may affect cell wall biosynthesis and membrane permeability, and showed deleterious effects against C. albicans biofilms. Copyright © 2016. Published by Elsevier Ltd.

  15. A Flexible Optical pH Sensor Based on Polysulfone Membranes Coated with pH-Responsive Polyaniline Nanofibers.

    PubMed

    Abu-Thabit, Nedal; Umar, Yunusa; Ratemi, Elaref; Ahmad, Ayman; Ahmad Abuilaiwi, Faraj

    2016-06-27

    A new optical pH sensor based on polysulfone (PSU) and polyaniline (PANI) was developed. A transparent and flexible PSU membrane was employed as a support. The electrically conductive and pH-responsive PANI was deposited onto the membrane surface by in situ chemical oxidative polymerization (COP). The absorption spectra of the PANI-coated PSU membranes exhibited sensitivity to pH changes in the range of 4-12, which allowed for designing a dual wavelength pH optical sensor. The performance of the membranes was assessed by measuring their response starting from high pH and going down to low pH, and vice versa. It was found that it is necessary to precondition the sensor layers before each measurement due to the slight hysteresis observed during forward and backward pH titrations. PSU membranes with polyaniline coating thicknesses in the range of ≈100-200 nm exhibited fast response times of <4 s, which are attributed to the porous, rough and nanofibrillar morphology of the polyaniline coating. The fabricated pH sensor was characterized by a sigmoidal response (R² = 0.997) which allows for pH determination over a wide dynamic range. All membranes were stable for a period of more than six months when stored in 1 M HCl solution. The reproducibility of the fabricated optical pH sensors was found to be <0.02 absorption units after one month storage in 1 M HCl solution. The performance of the optical pH sensor was tested and the obtained pH values were compared with the results obtained using a pH meter device.

  16. Anion- or Cation-Exchange Membranes for NaBH4/H2O2 Fuel Cells?

    PubMed

    Sljukić, Biljana; Morais, Ana L; Santos, Diogo M F; Sequeira, César A C

    2012-07-19

    Direct borohydride fuel cells (DBFC), which operate on sodium borohydride (NaBH4) as the fuel, and hydrogen peroxide (H2O2) as the oxidant, are receiving increasing attention. This is due to their promising use as power sources for space and underwater applications, where air is not available and gas storage poses obvious problems. One key factor to improve the performance of DBFCs concerns the type of separator used. Both anion- and cation-exchange membranes may be considered as potential separators for DBFC. In the present paper, the effect of the membrane type on the performance of laboratory NaBH4/H2O2 fuel cells using Pt electrodes is studied at room temperature. Two commercial ion-exchange membranes from Membranes International Inc., an anion-exchange membrane (AMI-7001S) and a cation-exchange membrane (CMI-7000S), are tested as ionic separators for the DBFC. The membranes are compared directly by the observation and analysis of the corresponding DBFC's performance. Cell polarization, power density, stability, and durability tests are used in the membranes' evaluation. Energy densities and specific capacities are estimated. Most tests conducted, clearly indicate a superior performance of the cation-exchange membranes over the anion-exchange membrane. The two membranes are also compared with several other previously tested commercial membranes. For long term cell operation, these membranes seem to outperform the stability of the benchmark Nafion membranes but further studies are still required to improve their instantaneous power load.

  17. Detection of Micrococcus luteus biofilm formation in microfluidic environments by pH measurement using an ion-sensitive field-effect transistor.

    PubMed

    Matsuura, Koji; Asano, Yuka; Yamada, Akira; Naruse, Keiji

    2013-02-18

    Biofilm formation in microfluidic channels is difficult to detect because sampling volumes are too small for conventional turbidity measurements. To detect biofilm formation, we used an ion-sensitive field-effect transistor (ISFET) measurement system to measure pH changes in small volumes of bacterial suspension. Cells of Micrococcus luteus (M. luteus) were cultured in polystyrene (PS) microtubes and polymethylmethacrylate (PMMA)-based microfluidic channels laminated with polyvinylidene chloride. In microtubes, concentrations of bacteria and pH in the suspension were analyzed by measuring turbidity and using an ISFET sensor, respectively. In microfluidic channels containing 20 μL of bacterial suspension, we measured pH changes using the ISFET sensor and monitored biofilm formation using a microscope. We detected acidification and alkalinization phases of M. luteus from the ISFET sensor signals in both microtubes and microfluidic channels. In the alkalinization phase, after 2 day culture, dense biofilm formation was observed at the bottom of the microfluidic channels. In this study, we used an ISFET sensor to detect biofilm formation in clinical and industrial microfluidic environments by detecting alkalinization of the culture medium. 

  18. Detection of pH and Enzyme-Free H2O2 Sensing Mechanism by Using GdO x Membrane in Electrolyte-Insulator-Semiconductor Structure.

    PubMed

    Kumar, Pankaj; Maikap, Siddheswar; Qiu, Jian-Tai; Jana, Surajit; Roy, Anisha; Singh, Kanishk; Cheng, Hsin-Ming; Chang, Mu-Tung; Mahapatra, Rajat; Chiu, Hsien-Chin; Yang, Jer-Ren

    2016-12-01

    A 15-nm-thick GdO x membrane in an electrolyte-insulator-semiconductor (EIS) structure shows a higher pH sensitivity of 54.2 mV/pH and enzyme-free hydrogen peroxide (H2O2) detection than those of the bare SiO2 and 3-nm-thick GdO x membranes for the first time. Polycrystalline grain and higher Gd content of the thicker GdO x films are confirmed by transmission electron microscopy (TEM) and X-ray photo-electron spectroscopy (XPS), respectively. In a thicker GdO x membrane, polycrystalline grain has lower energy gap and Gd(2+) oxidation states lead to change Gd(3+) states in the presence of H2O2, which are confirmed by electron energy loss spectroscopy (EELS). The oxidation/reduction (redox) properties of thicker GdO x membrane with higher Gd content are responsible for detecting H2O2 whereas both bare SiO2 and thinner GdO x membranes do not show sensing. A low detection limit of 1 μM is obtained due to strong catalytic activity of Gd. The reference voltage shift increases with increase of the H2O2 concentration from 1 to 200 μM owing to more generation of Gd(3+) ions, and the H2O2 sensing mechanism has been explained as well.

  19. [Inhibition analysis of resveratrol against Vibrio parahaemolyticus biofilm based on RNA-Seq technology].

    PubMed

    Zhang, Fang; Zhu, Junli; Feng, Lifang

    2016-05-04

    Food phytochemicals as biofilm inhibitor of pathogens have been highlighted. Our study aimed to investigate the effects of resveratrol on biofilm formation of an aquatic pathogen Vibrio parahaemolyticus, and to elucidate the important regulatory genes. In the subinhibitory concentrations, the inhibition of resveratrol aganist biofilm and exopolysaccharides of V. parahaemolyticus was detected, and the differentially expressed genes were analyzed based on RNA-Seq. Four genes involved in biofilm formation was validated by qRT-PCR. The minimum inhibitory concentration of resveratrol against V. parahaemolyticus was 20 μg/mL. Resveratrol at the subinhibitory concentration of 5 μg/mL and 10 μg/mL significantly decreased the biofilm development and exopolysaccharides production in V. parahaemolyticus (P < 0.05). Scanning electron microscopy micrographs showed a significant reduction of adherence and extracellular polymeric substances. RNA-Seq analysis revealed that 22.6% up-regulated and 77.4% down-regulated gene (P < 0.05) after treatment by 10 μg/mL resveratrol among 106 differential gene expressions. These differential genes in V. parahaemolyticu focused on 7 metabolic pathways, and 14 genes involved in biofilm formation were down-regulated by resveratrol, such as outer membrane protein (W, YedS, OmpK), quorum sensing (LuxS), flagellin (FlaA), fimbrial assembly protein (PilQ), hemolysin secreted protein. qRT-PCR confirmed that the expressions of luxS, trh, tlh and flaA, was significantly repressed in the presence of resveratrol, which was consistent with transcriptomics data. Inhibitory activity of resveratrol on biofilm formation was assicated with multiple genes and diverse cellular processes in V. parahaemolyticus. These findings suggest that resveratrol would disturb various metabolic pathways, particularly quorum sensing system, adhesion process and membrane proteins secretion, resulting in inhibition of attachment and biofilm development. The present work

  20. From biofilm ecology to reactors: a focused review.

    PubMed

    Boltz, Joshua P; Smets, Barth F; Rittmann, Bruce E; van Loosdrecht, Mark C M; Morgenroth, Eberhard; Daigger, Glen T

    2017-04-01

    Biofilms are complex biostructures that appear on all surfaces that are regularly in contact with water. They are structurally complex, dynamic systems with attributes of primordial multicellular organisms and multifaceted ecosystems. The presence of biofilms may have a negative impact on the performance of various systems, but they can also be used beneficially for the treatment of water (defined herein as potable water, municipal and industrial wastewater, fresh/brackish/salt water bodies, groundwater) as well as in water stream-based biological resource recovery systems. This review addresses the following three topics: (1) biofilm ecology, (2) biofilm reactor technology and design, and (3) biofilm modeling. In so doing, it addresses the processes occurring in the biofilm, and how these affect and are affected by the broader biofilm system. The symphonic application of a suite of biological methods has led to significant advances in the understanding of biofilm ecology. New metabolic pathways, such as anaerobic ammonium oxidation (anammox) or complete ammonium oxidation (comammox) were first observed in biofilm reactors. The functions, properties, and constituents of the biofilm extracellular polymeric substance matrix are somewhat known, but their exact composition and role in the microbial conversion kinetics and biochemical transformations are still to be resolved. Biofilm grown microorganisms may contribute to increased metabolism of micro-pollutants. Several types of biofilm reactors have been used for water treatment, with current focus on moving bed biofilm reactors, integrated fixed-film activated sludge, membrane-supported biofilm reactors, and granular sludge processes. The control and/or beneficial use of biofilms in membrane processes is advancing. Biofilm models have become essential tools for fundamental biofilm research and biofilm reactor engineering and design. At the same time, the divergence between biofilm modeling and biofilm reactor

  1. Inhibition of Escherichia coli O157:H7 on stainless steel using Pseudomonas veronii biofilms.

    PubMed

    Kim, Y; Kim, H; Beuchat, L R; Ryu, J-H

    2018-05-01

    We produced a Pseudomonas veronii biofilm on the surface of a stainless steel that is inhibitory to Escherichia coli O157:H7. Pseudomonas veronii strain KACC 81051BP, isolated from lettuce, readily formed biofilm on the surface of stainless steel coupons (SSCs) immersed in tryptic soy broth at 25°C. Cells showed significantly (P ≤ 0·05) enhanced tolerance to desiccation stress (43% relative humidity (RH)) and retained antimicrobial activity against E. coli O157:H7. The number of E. coli O157:H7 (control; 4·1 ± 0·1 log CFU per coupon) on sterile SSCs decreased to 2·7 ± 0·2 log CFU per coupon after exposure to 43% RH at 25°C for 48 h, while the population of E. coli O157:H7 (4·1 ± 0·0 log CFU per coupon) on SSCs containing P. veronii biofilm decreased to below the theoretical detection limit (1·5 log CFU per coupon) within 24 h. The antimicrobial biofilm produced on stainless steel may have application in preventing cross-contamination by E. coli O157:H7 on other abiotic surfaces in food-contact environments. The presence of Escherichia coli O157:H7 on environmental surfaces of food manufacturing, transportation and storage facilities is a significant food safety concern because it can result in cross-contamination of food products. In this study, we developed a Pseudomonas veronii biofilm on the surface of a stainless steel that inhibits the growth of E. coli O157:H7. Since P. veronii in biofilm resists desiccation, it provides persistent antimicrobial activity. Information presented here provides novel and practical insights to developing biological strategies to inactivate E. coli O157:H7 on diverse surfaces in food processing and handling environments. © 2018 The Society for Applied Microbiology.

  2. Model-Based Feasibility Assessment of Membrane Biofilm Reactor to Achieve Simultaneous Ammonium, Dissolved Methane, and Sulfide Removal from Anaerobic Digestion Liquor

    PubMed Central

    Chen, Xueming; Liu, Yiwen; Peng, Lai; Yuan, Zhiguo; Ni, Bing-Jie

    2016-01-01

    In this study, the membrane biofilm reactor (MBfR) is proposed to achieve simultaneous removal of ammonium, dissolved methane, and sulfide from main-stream and side-stream anaerobic digestion liquors. To avoid dissolved methane stripping, oxygen is introduced through gas-permeable membranes, which also from the substratum for the growth of a biofilm likely comprising ammonium oxidizing bacteria (AOB), anaerobic ammonium oxidation (Anammox) bacteria, denitrifying anaerobic methane oxidation (DAMO) microorganisms, aerobic methane oxidizing bacteria (MOB), and sulfur oxidizing bacteria (SOB). A mathematical model is developed and applied to assess the feasibility of such a system and the associated microbial community structure under different operational conditions. The simulation studies demonstrate the feasibility of achieving high-level (>97.0%), simultaneous removal of ammonium, dissolved methane, and sulfide in the MBfRs from both main-stream and side-stream anaerobic digestion liquors through adjusting the influent surface loading (or hydraulic retention time (HRT)) and the oxygen surface loading. The optimal HRT was found to be inversely proportional to the corresponding oxygen surface loading. Under the optimal operational conditions, AOB, DAMO bacteria, MOB, and SOB dominate the biofilm of the main-stream MBfR, while AOB, Anammox bacteria, DAMO bacteria, and SOB coexist in the side-stream MBfR to remove ammonium, dissolved methane, and sulfide simultaneously. PMID:27112502

  3. Photosynthetic solar cell using nanostructured proton exchange membrane for microbial biofilm prevention.

    PubMed

    Lee, Dong Hyun; Oh, Hwa Jin; Bai, Seoung Jae; Song, Young Seok

    2014-06-24

    Unwanted biofilm formation has a detrimental effect on bioelectrical energy harvesting in microbial cells. This issue still needs to be solved for higher power and longer durability and could be resolved with the help of nanoengineering in designing and manufacturing. Here, we demonstrate a photosynthetic solar cell (PSC) that contains a nanostructure to prevent the formation of biofilm by micro-organisms. Nanostructures were fabricated using nanoimprint lithography, where a film heater array system was introduced to precisely control the local wall temperature. To understand the heat and mass transfer phenomena behind the manufacturing and energy harvesting processes of PSC, we carried out a numerical simulation and experimental measurements. It revealed that the nanostructures developed on the proton exchange membrane enable PSC to produce enhanced output power due to the retarded microbial attachment on the Nafion membrane. We anticipate that this strategy can provide a pathway where PSC can ensure more renewable, sustainable, and efficient energy harvesting performance.

  4. The Interaction of CuS and Halothiobacillus HT1 Biofilm in Microscale Using Synchrotron Radiation-Based Techniques

    PubMed Central

    Lin, Huirong; Chen, Guangcun; Zhu, Shenhai; Chen, Yingxu; Chen, Dongliang; Xu, Wei; Yu, Xiaohan; Shi, Jiyan

    2013-01-01

    In order to investigate the microbe-mineral interaction in the micro scale, spatial distribution and speciation of Cu and S in Halothiobacillus HT1 biofilm formed on a CuS surface was examined using synchrotron-based X-ray techniques. Confocal laser scanning microscope (CLSM) results indicated that Halothiobacillus HT1 biofilm formation gave rise to distinct chemical and redox gradients, leading to diverse niches in the biofilm. Live cells were distributed at the air-biofilm and membrane-biofilm interface. CuS was oxidized by Halothiobacillus HT1 biofilm, and copper penetrated into the biofilm. Sulfide was oxidized to cysteine (77.3%), sulfite (3.8%) and sulfonate (18.9%). Cu-cysteine-like species were involved in the copper homeostasis. These results significantly improve our understanding of the interfacial properties of the biofilm-mineral interface. PMID:23708108

  5. Anion- or Cation-Exchange Membranes for NaBH4/H2O2 Fuel Cells?

    PubMed Central

    Šljukić, Biljana; Morais, Ana L.; Santos, Diogo M. F.; Sequeira, César A. C.

    2012-01-01

    Direct borohydride fuel cells (DBFC), which operate on sodium borohydride (NaBH4) as the fuel, and hydrogen peroxide (H2O2) as the oxidant, are receiving increasing attention. This is due to their promising use as power sources for space and underwater applications, where air is not available and gas storage poses obvious problems. One key factor to improve the performance of DBFCs concerns the type of separator used. Both anion- and cation-exchange membranes may be considered as potential separators for DBFC. In the present paper, the effect of the membrane type on the performance of laboratory NaBH4/H2O2 fuel cells using Pt electrodes is studied at room temperature. Two commercial ion-exchange membranes from Membranes International Inc., an anion-exchange membrane (AMI-7001S) and a cation-exchange membrane (CMI-7000S), are tested as ionic separators for the DBFC. The membranes are compared directly by the observation and analysis of the corresponding DBFC’s performance. Cell polarization, power density, stability, and durability tests are used in the membranes’ evaluation. Energy densities and specific capacities are estimated. Most tests conducted, clearly indicate a superior performance of the cation-exchange membranes over the anion-exchange membrane. The two membranes are also compared with several other previously tested commercial membranes. For long term cell operation, these membranes seem to outperform the stability of the benchmark Nafion membranes but further studies are still required to improve their instantaneous power load. PMID:24958292

  6. Structural Basis for Translocation of a Biofilm-supporting Exopolysaccharide across the Bacterial Outer Membrane.

    PubMed

    Wang, Yan; Andole Pannuri, Archana; Ni, Dongchun; Zhou, Haizhen; Cao, Xiou; Lu, Xiaomei; Romeo, Tony; Huang, Yihua

    2016-05-06

    The partially de-N-acetylated poly-β-1,6-N-acetyl-d-glucosamine (dPNAG) polymer serves as an intercellular biofilm adhesin that plays an essential role for the development and maintenance of integrity of biofilms of diverse bacterial species. Translocation of dPNAG across the bacterial outer membrane is mediated by a tetratricopeptide repeat-containing outer membrane protein, PgaA. To understand the molecular basis of dPNAG translocation, we determined the crystal structure of the C-terminal transmembrane domain of PgaA (residues 513-807). The structure reveals that PgaA forms a 16-strand transmembrane β-barrel, closed by four loops on the extracellular surface. Half of the interior surface of the barrel that lies parallel to the translocation pathway is electronegative, suggesting that the corresponding negatively charged residues may assist the secretion of the positively charged dPNAG polymer. In vivo complementation assays in a pgaA deletion bacterial strain showed that a cluster of negatively charged residues proximal to the periplasm is necessary for biofilm formation. Biochemical analyses further revealed that the tetratricopeptide repeat domain of PgaA binds directly to the N-deacetylase PgaB and is critical for biofilm formation. Our studies support a model in which the positively charged PgaB-bound dPNAG polymer is delivered to PgaA through the PgaA-PgaB interaction and is further targeted to the β-barrel lumen of PgaA potentially via a charge complementarity mechanism, thus priming the translocation of dPNAG across the bacterial outer membrane. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  7. Evaluation of adhesive and anti-adhesive properties of Pseudomonas aeruginosa biofilms and their inhibition by herbal plants

    PubMed Central

    Zameer, Farhan; MS, Rukmangada; Chauhan, Jyoti Bala; Khanum, Shaukath Ara; Kumar, Pramod; Devi, Aishwarya Tripurasundari; MN, Nagendra Prasad; BL, Dhananjaya

    2016-01-01

    Background and Objectives: Adhesion and colonization are prerequisites for the establishment of bacterial pathogenesis. The biofilm development of Pseudomonas aeruginosa was assessed on adhesive surfaces like dialysis membrane, stainless steel, glass and polystyrene. Materials and Methods: Microtiter plate biofilm assay was performed to assess the effect of nutrient medium and growth parameters of P. aeruginosa. Further, its growth on adhesive surfaces namely hydrophilic (dialysis membrane) and hydrophobic (polystyrene plate, square glass and stainless steel coupon) was assessed. The exopolysaccharide (EPS) was quantified using ruthenium red microplate assay and microscopic analysis was used to observe P. aeruginosa biofilm architecture. The anti-biofilm activity of herbal extracts on mature P. aeruginosa was performed. Results: The formation of large scale biofilms on dialysis membrane for 72 h was proved to be the best surface. In microscopic studies, very few exopolysaccaride fibrils, indicating a rather loose matrix was observed at 48 h. Further, thick exopolysaccaride, indicated higher adhesive properties at 72 h which is evident from ruthenium red staining. Among the plant extract used, Justicia wynaadensis leaf and Aristolochia indica (Eswari) root extract showed significant reduction of anti-biofilm activity of 0.178 OD and 0.192 OD in inhibiting mature biofilms at 0.225 OD respectively, suggesting the possible use of these extracts as efficient anti-adhesive and biofilm-disrupting agents with potential applications in controlling biofilms on surfaces. Conclusion: Our study facilitates better understanding in the development of P. aeruginosa biofilms on different food processing and clinical surfaces ultimately taking care of food safety and hygiene. PMID:27307976

  8. "Twin copper source" growth of metal-organic framework membrane: Cu(3)(BTC)(2) with high permeability and selectivity for recycling H(2).

    PubMed

    Guo, Hailing; Zhu, Guangshan; Hewitt, Ian J; Qiu, Shilun

    2009-02-11

    In this communication, the copper net supported Cu(3)(BTC)(2) membranes have been successfully synthesized by means of a "twin copper source" technique. Separation studies on gaseous mixtures (H(2)/CO(2), H(2)/CH(4), and H(2)/N(2)) using the membrane revealed that the membrane possesses high permeability and selectivity for H(2) over CO(2), N(2), and CH(4). Compared with the conventional zeolite membranes, the copper net supported Cu(3)(BTC)(2) membrane exhibited high permeation flux in gas separation. Such highly efficient copper net supported Cu(3)(BTC)(2) membranes could be used to separate, recycle, and reuse H(2) exhausted from steam reforming natural gas.

  9. Effect of growth temperature, surface type and incubation time on the resistance of Staphylococcus aureus biofilms to disinfectants.

    PubMed

    Abdallah, Marwan; Chataigne, Gabrielle; Ferreira-Theret, Pauline; Benoliel, Corinne; Drider, Djamel; Dhulster, Pascal; Chihib, Nour-Eddine

    2014-03-01

    The goal of this study was to investigate the effect of the environmental conditions such as the temperature change, incubation time and surface type on the resistance of Staphylococcus aureus biofilms to disinfectants. The antibiofilm assays were performed against biofilms grown at 20 °C, 30 °C and 37 °C, on the stainless steel and polycarbonate, during 24 and 48 h. The involvement of the biofilm matrix and the bacterial membrane fluidity in the resistance of sessile cells were investigated. Our results show that the efficiency of disinfectants was dependent on the growth temperature, the surface type and the disinfectant product. The increase of growth temperature from 20 °C to 37 °C, with an incubation time of 24 h, increased the resistance of biofilms to cationic antimicrobials. This change of growth temperature did not affect the major content of the biofilm matrix, but it decreased the membrane fluidity of sessile cells through the increase of the anteiso-C19 relative amount. The increase of the biofilm resistance to disinfectants, with the rise of the incubation time, was dependent on both growth temperature and disinfectant product. The increase of the biofilm age also promoted increases in the matrix production and the membrane fluidity of sessile cells. The resistance of S. aureus biofilm seems to depend on the environment of the biofilm formation and involves both extracellular matrix and membrane fluidity of sessile cells. Our study represents the first report describing the impact of environmental conditions on the matrix production, sessile cells membrane fluidity and resistance of S. aureus biofilms to disinfectants.

  10. Development of an in vitro Assay, Based on the BioFilm Ring Test®, for Rapid Profiling of Biofilm-Growing Bacteria

    PubMed Central

    Di Domenico, Enea G.; Toma, Luigi; Provot, Christian; Ascenzioni, Fiorentina; Sperduti, Isabella; Prignano, Grazia; Gallo, Maria T.; Pimpinelli, Fulvia; Bordignon, Valentina; Bernardi, Thierry; Ensoli, Fabrizio

    2016-01-01

    Microbial biofilm represents a major virulence factor associated with chronic and recurrent infections. Pathogenic bacteria embedded in biofilms are highly resistant to environmental and chemical agents, including antibiotics and therefore difficult to eradicate. Thus, reliable tests to assess biofilm formation by bacterial strains as well as the impact of chemicals or antibiotics on biofilm formation represent desirable tools for a most effective therapeutic management and microbiological risk control. Current methods to evaluate biofilm formation are usually time-consuming, costly, and hardly applicable in the clinical setting. The aim of the present study was to develop and assess a simple and reliable in vitro procedure for the characterization of biofilm-producing bacterial strains for future clinical applications based on the BioFilm Ring Test® (BRT) technology. The procedure developed for clinical testing (cBRT) can provide an accurate and timely (5 h) measurement of biofilm formation for the most common pathogenic bacteria seen in clinical practice. The results gathered by the cBRT assay were in agreement with the traditional crystal violet (CV) staining test, according to the κ coefficient test (κ = 0.623). However, the cBRT assay showed higher levels of specificity (92.2%) and accuracy (88.1%) as compared to CV. The results indicate that this procedure offers an easy, rapid and robust assay to test microbial biofilm and a promising tool for clinical microbiology. PMID:27708625

  11. Salvianolic acid B protects hepatocytes from H2O2 injury by stabilizing the lysosomal membrane.

    PubMed

    Yan, Xiao-Feng; Zhao, Pei; Ma, Dong-Yan; Jiang, Yi-Lu; Luo, Jiao-Jiao; Liu, Liu; Wang, Xiao-Ling

    2017-08-07

    To investigate the capability of salvianolic acid B (Sal B) to protect hepatocytes from hydrogen peroxide (H 2 O 2 )/carbon tetrachloride (CCl 4 )-induced lysosomal membrane permeabilization. Cell Counting Kit-8 assay was used to measure cell viability. Apoptosis and death were assayed through flow cytometry. BrdU incorporation was used to detect cell proliferation. Serum alanine aminotransferase activity and liver malondialdehyde (MDA) content were measured. Liver histopathological changes were evaluated using hematoxylin-eosin staining. Lysosomal membrane permeability was detected with LysoTracker Green-labeled probes and acridine orange staining. The levels of protein carbonyl content (PCC), cathepsins (Cat)B/D, and lysosome-associated membrane protein 1 (LAMP1) were evaluated through western blotting. Cytosol CatB activity analysis was performed with chemiluminescence detection. The mRNA level of LAMP1 was evaluated through quantitative real-time polymerase chain reaction. Results indicated that H 2 O 2 induced cell injury/death. Sal B attenuated H 2 O 2 -induced cell apoptosis and death, restored the inhibition of proliferation, decreased the amount of PCC, and stabilized the lysosome membrane by increasing the LAMP1 protein level and antagonizing CatB/D leakage into the cytosol. CCl 4 also triggered hepatocyte death. Furthermore, Sal B effectively rescued hepatocytes by increasing LAMP1 expression and by reducing lysosomal enzyme translocation to the cytosol. Sal B protected mouse embryonic hepatocytes from H 2 O 2 /CCl 4 -induced injury/death by stabilizing the lysosomal membrane.

  12. Effect of UV on De-NOx performance and microbial community of a hybrid catalytic membrane biofilm reactor

    NASA Astrophysics Data System (ADS)

    Chen, Zhouyang; Huang, Zhensha; He, Yiming; Xiao, Xiaoliang; Wei, Zaishan

    2018-02-01

    The hybrid membrane catalytic biofilm reactor provides a new way of flue gas denitration. However, the effects of UV on denitrification performance, microbial community and microbial nitrogen metabolism are still unknown. In this study, the effects of UV on deNO x performance, nitrification and denitrification, microbial community and microbial nitrogen metabolism of a bench scale N-TiO2/PSF hybrid catalytic membrane biofilm reactor (HCMBR) were evaluated. The change from nature light to UV in the HCMBR leads to the fall of NO removal efficiency of HCMBR from 92.8% to 81.8%. UV affected the microbial community structure, but did not change microbial nitrogen metabolism, as shown by metagenomics sequencing method. Some dominant phyla, such as Gammaproteobacteria, Bacteroidetes, Firmicutes, Actinobacteria, and Alphaproteobacteria, increased in abundance, whereas others, such as Proteobacteria and Betaproteobacteria, decreased. There were nitrification, denitrification, nitrogen fixation, and organic nitrogen metabolism in the HCMBR.

  13. CuSO4/H2O2-Triggered Polydopamine/Poly(sulfobetaine methacrylate) Coatings for Antifouling Membrane Surfaces.

    PubMed

    Zhang, Chao; Li, Hao-Nan; Du, Yong; Ma, Meng-Qi; Xu, Zhi-Kang

    2017-02-07

    Mussel-inspired polydopamine (PDA) coatings have been broadly exploited for constructing functional membrane surfaces. One-step codeposition of PDA with antifouling polymers, especially zwitterionic polymers, has been regarded as a promising strategy for fabricating antifouling membrane surfaces. However, one challenge is that the codeposition is usually a slow process over 10 h or even several days. Herein, we report that CuSO 4 /H 2 O 2 is able to notably accelerate the codeposition process of PDA with poly(sulfobetaine methacrylate) (PSBMA). In our case, PSBMA is facilely anchored to the polypropylene microporous membrane (PPMM) surfaces within 1 h with the assistance of PDA because of its strong interfacial adhesion. The PDA/PSBMA-coated PPMMs show excellent surface hydrophilicity, high water permeation flux (7506 ± 528 L/m 2 ·h at 0.1 MPa), and an outstanding antifouling property. Moreover, the antifouling property is maintained after the membranes are treated with acid and alkali solutions as well as organic solvents. To recap, it provides a facile, universal, and time-saving strategy for exploiting high-efficiency and durable antifouling membrane surfaces.

  14. Biofilm-specific extracellular matrix proteins of non-typeable Haemophilus influenzae

    PubMed Central

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

    2014-01-01

    Non-typeable 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 hr and 96 hr NTHi biofilms contained polysaccharides and proteinaceous components as detected by NMR and 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 hr biofilms, two were found only in 96 hr biofilms, and fifteen were present in the ECM of both 24 hr and 96 hr NTHi biofilms. All proteins identified were either associated with bacterial membranes or were 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. PMID:24942343

  15. Total electron acceptor loading and composition affect hexavalent uranium reduction and microbial community structure in a membrane biofilm reactor.

    PubMed

    Ontiveros-Valencia, Aura; Zhou, Chen; Ilhan, Zehra Esra; de Saint Cyr, Louis Cornette; Krajmalnik-Brown, Rosa; Rittmann, Bruce E

    2017-11-15

    Molecular microbiology tools (i.e., 16S rDNA gene sequencing) were employed to elucidate changes in the microbial community structure according to the total electron acceptor loading (controlled by influent flow rate and/or medium composition) in a H 2 -based membrane biofilm reactor evaluated for removal of hexavalent uranium. Once nitrate, sulfate, and dissolved oxygen were replaced by U(VI) and bicarbonate and the total acceptor loading was lowered, slow-growing bacteria capable of reducing U(VI) to U(IV) dominated in the biofilm community: Replacing denitrifying bacteria Rhodocyclales and Burkholderiales were spore-producing Clostridiales and Natranaerobiales. Though potentially competing for electrons with U(VI) reducers, homo-acetogens helped attain steady U(VI) reduction, while methanogenesis inhibited U(VI) reduction. U(VI) reduction was reinstated through suppression of methanogenesis by addition of bromoethanesulfonate or by competition from SRB when sulfate was re-introduced. Predictive metagenome analysis further points out community changes in response to alterations in the electron-acceptor loading: Sporulation and homo-acetogenesis were critical factors for strengthening stable microbial U(VI) reduction. This study documents that sporulation was important to long-term U(VI) reduction, whether or not microorganisms that carry out U(VI) reduction mediated by cytochrome c 3 , such as SRB and ferric-iron-reducers, were inhibited. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. 220D-F2 from Rubus ulmifolius kills Streptococcus pneumoniae planktonic cells and pneumococcal biofilms.

    PubMed

    Talekar, Sharmila J; Chochua, Sopio; Nelson, Katie; Klugman, Keith P; Quave, Cassandra L; Vidal, Jorge E

    2014-01-01

    Streptococcus pneumoniae (pneumococcus) forms organized biofilms to persist in the human nasopharynx. This persistence allows the pneumococcus to produce severe diseases such as pneumonia, otitis media, bacteremia and meningitis that kill nearly a million children every year. While bacteremia and meningitis are mediated by planktonic pneumococci, biofilm structures are present during pneumonia and otitis media. The global emergence of S. pneumoniae strains resistant to most commonly prescribed antibiotics warrants further discovery of alternative therapeutics. The present study assessed the antimicrobial potential of a plant extract, 220D-F2, rich in ellagic acid, and ellagic acid derivatives, against S. pneumoniae planktonic cells and biofilm structures. Our studies first demonstrate that, when inoculated together with planktonic cultures, 220D-F2 inhibited the formation of pneumococcal biofilms in a dose-dependent manner. As measured by bacterial counts and a LIVE/DEAD bacterial viability assay, 100 and 200 µg/ml of 220D-F2 had significant bactericidal activity against pneumococcal planktonic cultures as early as 3 h post-inoculation. Quantitative MIC's, whether quantified by qPCR or dilution and plating, showed that 80 µg/ml of 220D-F2 completely eradicated overnight cultures of planktonic pneumococci, including antibiotic resistant strains. When preformed pneumococcal biofilms were challenged with 220D-F2, it significantly reduced the population of biofilms 3 h post-inoculation. Minimum biofilm inhibitory concentration (MBIC)50 was obtained incubating biofilms with 100 µg/ml of 220D-F2 for 3 h and 6 h of incubation. 220D-F2 also significantly reduced the population of pneumococcal biofilms formed on human pharyngeal cells. Our results demonstrate potential therapeutic applications of 220D-F2 to both kill planktonic pneumococcal cells and disrupt pneumococcal biofilms.

  17. A simple optode based method for imaging O2 distribution and dynamics in tap water biofilms.

    PubMed

    Staal, M; Prest, E I; Vrouwenvelder, J S; Rickelt, L F; Kühl, M

    2011-10-15

    A ratiometric luminescence intensity imaging approach is presented, which enables spatial O2 measurements in biofilm reactors with transparent planar O2 optodes. Optodes consist of an O2 sensitive luminescent dye immobilized in a 1-10 μm thick polymeric layer on a transparent carrier, e.g. a glass window. The method is based on sequential imaging of the O2 dependent luminescence intensity, which are subsequently normalized with luminescent intensity images recorded under anoxic conditions. We present 2-dimensional O2 distribution images at the base of a tap water biofilm measured with the new ratiometric method and compare the results with O2 distribution images obtained in the same biofilm reactor with luminescence lifetime imaging. Using conventional digital cameras, such simple normalized luminescence intensity imaging can yield images of 2-dimensional O2 distributions with a high signal-to-noise ratio and spatial resolution comparable or even surpassing those obtained with expensive and complex luminescence lifetime imaging systems. The method can be applied to biofilm growth incubators allowing intermittent experimental shifts to anoxic conditions or in systems, in which the O2 concentration is depleted during incubation. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. Apple Flavonoid Phloretin Inhibits Escherichia coli O157:H7 Biofilm Formation and Ameliorates Colon Inflammation in Rats ▿ †

    PubMed Central

    Lee, Jin-Hyung; Regmi, Sushil Chandra; Kim, Jung-Ae; Cho, Moo Hwan; Yun, Hyungdon; Lee, Chang-Soo; Lee, Jintae

    2011-01-01

    Pathogenic biofilms have been associated with persistent infections due to their high resistance to antimicrobial agents, while commensal biofilms often fortify the host's immune system. Hence, controlling biofilm formation of both pathogenic bacteria and commensal bacteria is important in bacterium-related diseases. We investigated the effect of plant flavonoids on biofilm formation of enterohemorrhagic Escherichia coli O157:H7. The antioxidant phloretin, which is abundant in apples, markedly reduced E. coli O157:H7 biofilm formation without affecting the growth of planktonic cells, while phloretin did not harm commensal E. coli K-12 biofilms. Also, phloretin reduced E. coli O157:H7 attachment to human colon epithelial cells. Global transcriptome analyses revealed that phloretin repressed toxin genes (hlyE and stx2), autoinducer-2 importer genes (lsrACDBF), curli genes (csgA and csgB), and dozens of prophage genes in E. coli O157:H7 biofilm cells. Electron microscopy confirmed that phloretin reduced fimbria production in E. coli O157:H7. Also, phloretin suppressed the tumor necrosis factor alpha-induced inflammatory response in vitro using human colonic epithelial cells. Moreover, in the rat model of colitis induced by trinitrobenzene sulfonic acid (TNBS), phloretin significantly ameliorated colon inflammation and body weight loss. Taken together, our results suggest that the antioxidant phloretin also acts as an inhibitor of E. coli O157:H7 biofilm formation as well as an anti-inflammatory agent in inflammatory bowel diseases without harming beneficial commensal E. coli biofilms. PMID:21930760

  19. Inhibition of Candida albicans Biofilm Formation by the Synthetic Lactoferricin Derived Peptide hLF1-11

    PubMed Central

    Morici, Paola; Fais, Roberta; Rizzato, Cosmeri

    2016-01-01

    The aim of this study was to evaluate the in vitro activity of the synthetic peptide hLF1-11 against biofilm produced by clinical isolates of Candida albicans with different fluconazole susceptibility. The antibiofilm activity of the peptide hLF1-11 was assessed in terms of reduction of biofilm cellular density, metabolic activity and sessile cell viability. The extent of morphogenesis in hLF1-11 treated and untreated biofilms was also investigated microscopically. Transcription levels of genes related to cell adhesion, hyphal development and extracellular matrix production were analysed by qRT-PCR in hLF1-11 treated and untreated biofilms. Exogenous dibutyryl-cAMP (db-cAMP) was used to rescue morphogenesis in cells exposed to the peptide. The results revealed that hLF1-11 exhibited an inhibitory effect on biofilm formation by all C. albicans isolates tested in a dose-dependent manner, regardless of their fluconazole susceptibility. Visual inspection of treated or untreated biofilm cells with an inverted microscope revealed a significant reduction in hyphal formation by hLF1-11 treated cells, as early as 3 hours of incubation. Moreover, hLF1-11 showed a reduced activity on preadherent cells. hLF1-11 induced the down-regulation of biofilm and hyphal-associated genes, which were predominantly regulated via the Ras1-cAMP-Efg1 pathway. Indeed, exogenous db-cAMP restored morphogenesis in hLF1-11 treated cells. The hLF1-11 peptide significantly inhibited biofilm formation by C. albicans mainly at early stages, interfering with biofilm cellular density and metabolic activity, and affected morphogenesis through the Ras1-cAMP-Efg1 pathway. Our findings provide the first evidence that hLF1-11 could represent a potential candidate for the prevention of biofilm formation by C. albicans. PMID:27902776

  20. Inhibition of Candida albicans Biofilm Formation by the Synthetic Lactoferricin Derived Peptide hLF1-11.

    PubMed

    Morici, Paola; Fais, Roberta; Rizzato, Cosmeri; Tavanti, Arianna; Lupetti, Antonella

    2016-01-01

    The aim of this study was to evaluate the in vitro activity of the synthetic peptide hLF1-11 against biofilm produced by clinical isolates of Candida albicans with different fluconazole susceptibility. The antibiofilm activity of the peptide hLF1-11 was assessed in terms of reduction of biofilm cellular density, metabolic activity and sessile cell viability. The extent of morphogenesis in hLF1-11 treated and untreated biofilms was also investigated microscopically. Transcription levels of genes related to cell adhesion, hyphal development and extracellular matrix production were analysed by qRT-PCR in hLF1-11 treated and untreated biofilms. Exogenous dibutyryl-cAMP (db-cAMP) was used to rescue morphogenesis in cells exposed to the peptide. The results revealed that hLF1-11 exhibited an inhibitory effect on biofilm formation by all C. albicans isolates tested in a dose-dependent manner, regardless of their fluconazole susceptibility. Visual inspection of treated or untreated biofilm cells with an inverted microscope revealed a significant reduction in hyphal formation by hLF1-11 treated cells, as early as 3 hours of incubation. Moreover, hLF1-11 showed a reduced activity on preadherent cells. hLF1-11 induced the down-regulation of biofilm and hyphal-associated genes, which were predominantly regulated via the Ras1-cAMP-Efg1 pathway. Indeed, exogenous db-cAMP restored morphogenesis in hLF1-11 treated cells. The hLF1-11 peptide significantly inhibited biofilm formation by C. albicans mainly at early stages, interfering with biofilm cellular density and metabolic activity, and affected morphogenesis through the Ras1-cAMP-Efg1 pathway. Our findings provide the first evidence that hLF1-11 could represent a potential candidate for the prevention of biofilm formation by C. albicans.

  1. Non-Tuberculous Mycobacteria multispecies biofilms in cystic fibrosis: development of an in vitro Mycobacterium abscessus and Pseudomonas aeruginosa dual species biofilm model.

    PubMed

    Rodríguez-Sevilla, Graciela; García-Coca, Marta; Romera-García, David; Aguilera-Correa, John Jairo; Mahíllo-Fernández, Ignacio; Esteban, Jaime; Pérez-Jorge, Concepción

    2018-04-01

    Lung disease in cystic fibrosis (CF) is characterized by the progressive colonization of the respiratory tract by different bacteria, which develop polymicrobial biofilms. In the past decades, there has been an increase in the number of CF patients infected with Non-Tuberculous Mycobacteria (NTM). Although Mycobacterium abscessus is the main NTM isolated globally, little is known about M. abscessus multispecies biofilm formation. In the present study we developed an in vitro model to study the phenotypic characteristics of biofilms formed by M. abscessus and Pseudomonas aeruginosa, a major pathogen in CF. For that purpose, dual species biofilms were grown on polycarbonate membranes with a fixed concentration of P. aeruginosa and different inoculums of M. abscessus. The biofilms were sampled at 24, 48, and 72 h and bacteria were quantified in specific media. The results revealed that the increasing initial concentration of M. abscessus in dual species biofilms had an effect on its population only at 24 and 48 h, whereas P. aeruginosa was not affected by the different concentrations used of M. abscessus. Time elapsed increased biofilm formation of both species, specially between 24 and 48 h. According to the results, the conditions to produce a mature dual species biofilm in which the relative species distribution remained stable were 72 h growth of the mixed microbial culture at a 1:1 ratio. A significant decrease in mycobacterial population in dual compared to single species biofilms was found, suggesting that P. aeruginosa has a negative influence on M. abscessus. Finally, in a proof of concept experiment, young and mature dual species biofilms were exposed to clarithromycin. Copyright © 2018 Elsevier GmbH. All rights reserved.

  2. Design of an optically stable pH sensor based on immobilization of Giemsa on triacetylcellulose membrane.

    PubMed

    Khodadoust, Saeid; Kouri, Narges Cham; Talebiyanpoor, Mohammad Sharif; Deris, Jamile; Pebdani, Arezou Amiri

    2015-12-01

    In this work a simple, inexpensive, and sensitive optical sensor based on triacetylcellulose membrane as solid support was developed by using immobilization of Giemsa indicator for pH measurement. In this method, the influence variables on the membrane performance including pH concentration of indicator, response time, ionic strength, and reversibility were investigated. At optimum values of all variables the response of optical pH sensor is linear in the pH range of 3.0-12.0. This optical sensor was produced through simultaneous binding of the Giemsa on the activated triacetylcellulose membrane which responded to the pH changes in a broader linear range within less than 2.0 min and suitable reproducibility (RSD<5%). Stability results showed that this sensor was stable after 6 months of storage in the water/ethanol (50:50, v/v) solution without any measurable divergence in response properties (less than 5% RSD). Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Progesterone, Inflammatory Cytokine (TNF-α), and Oxidative Stress (H2O2) Regulate Progesterone Receptor Membrane Component 1 Expression in Fetal Membrane Cells.

    PubMed

    Meng, Yan; Murtha, Amy P; Feng, Liping

    2016-09-01

    Progesterone receptor membrane component 1 (PGRMC1) is an important novel mediator of progesterone (P4) function in fetal membrane cells. We demonstrated previously that PGRMC1 is differentially expressed in fetal membranes among pregnancy subjects and diminished in preterm premature rupture of membrane subjects. In the current study, we aim to elucidate whether PGRMC1 expression is regulated by P4, tumor necrosis factor α (TNF-α), and H2O2 in fetal membrane cells. Primary cultured membrane cells were serum starved for 24 hours followed by treatments of P4, 17 hydroxyprogesterone caproate, and medroxyprogesterone 17 acetate (MPA) at 10(-7) mol/L with ethanol as vehicle control; TNF-α at 10, 20, and 50 ng/mL with phosphate-buffered saline (PBS) as control; and H2O2 at 10 and 100 μmol/L with culture media as control for 24, 48, and 72 hours. The messenger RNA (mRNA) and protein expression of PGRMC1 was quantified using polymerase chain reaction and Western blotting, respectively. We found that PGRMC1 protein expression was regulated by MPA, TNF-α, and H2O2 in a dose-dependent manner. This regulation is also specific to the type of cell (amnion, chorion, or decidua). The upregulation of PGRMC1 by MPA might be mediated through glucocorticoid receptor (GR) demonstrated using amnion and chorion cells model with GR knockdown by specific small interfering RNA transfection. The mRNA expression of PGRMC1 was decreased by H2O2 (100 μmol/L) treatment in amnion cells, which might ultimately result in downregulation of PGRMC1 protein as our data demonstrated. None of other treatments changed PGRMC1 mRNA level in these cells. We conclude that these stimuli act as regulatory factors of PGRMC1 in a cell-specific manner. © The Author(s) 2016.

  4. Evaluation of kenaf fibers as moving bed biofilm carriers in algal membrane photobioreactor.

    PubMed

    Derakhshan, Zahra; Mahvi, Amir Hossein; Ehrampoush, Mohammad Hassan; Ghaneian, Mohammad Taghi; Yousefinejad, Saeed; Faramarzian, Mohammad; Mazloomi, Seyed Mohammad; Dehghani, Mansooreh; Fallahzadeh, Hossein

    2018-05-15

    In this lab-scale study, the feasibility of using kenaf fibers as moving bed biofilm carriers in hybrid microalgal membrane photobioreactors (HMPBR) in organic matter and atrazine elimination from real secondary effluent was evaluated. For evaluating the kinetics of biofilm substrate consumption, an experimental model was proposed. Inoculation of wastewater samples with free carriers resulted in the greater removal of target pollutants. Removal efficiency of atrazine and chemical oxygen demand (COD) increased to 27% and 16%, with respect to the control, respectively. The total biomass accumulation in HMPBR exceeded 5g/L, and the microalgae tended to aggregate and attached to biofilm carriers. The removal efficiency of HMPBR improved significantly via inoculation of kenaf fiber carriers with bioremediation microalgal strains (p < 0.01). A lower stabilization ratio (VSS/TSS) was also recorded. The biomass in HMPBR included more lipids and carbohydrates. The results revealed that kenaf fibers could improve and upgrade the biological activity of different wastewater treatment applications, considering the great potential of biofilm carriers and their effluent quality. Copyright © 2018 Elsevier Inc. All rights reserved.

  5. The characteristics of extracellular polymeric substances and soluble microbial products in moving bed biofilm reactor-membrane bioreactor.

    PubMed

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

    2013-11-01

    The characteristics of extracellular polymeric substances (EPS) and soluble microbial products (SMP) in conventional membrane bioreactor (MBR) and in moving bed biofilm reactor-membrane bioreactors (MBBR-MBR) were investigated in long-term (170 days) experiments. The results showed that all reactors had high removal efficiency of ammonium and COD, despite very different fouling conditions. The MBBR-MBR with media fill ratio of 26.7% had much lower total membrane resistance and no obvious fouling were detected during the whole operation. In contrast, MBR and MBBR-MBR with lower and higher media fill experienced more significant fouling. Low fouling at optimum fill ratio may be due to the higher percentage of small molecular size (<1 kDa) and lower percentage of large molecular size (>100 kDa) of EPS and SMP in the reactor. The composition of EPS and SMP affected fouling due to different O-H bonds in hydroxyl functional groups, and less polysaccharides and lipids. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Effect of Holarrhena antidysentrica (Ha) and Andrographis paniculata (Ap) on the biofilm formation and cell membrane integrity of opportunistic pathogen Salmonella typhimurium.

    PubMed

    Tanwar, Ankit; Chawla, Raman; Chakotiya, Ankita Singh; Thakur, Pallavi; Goel, Rajeev; Basu, Mitra; Arora, Rajesh; Khan, Haider Ali

    2016-12-01

    Increasing occurrence of gastroenteritis outbreaks caused by food borne opportunistic microorganisms has become a major problem in food industry as well as in immunocompromised host. Antimicrobial agents are losing their efficacy due to increase in the microbial resistance. For such reasons, conventional treatment has become limited to manage the infections state. Need of the hour is to instigate the search for safer holistic alternatives. The present study was hence conducted to assess the antibiofilm effect and mode of action of aquo alcoholic extracts of Holarrhena antidysentrica (Ha) and Andrographis paniculata (Ap) against the Salmonella enterica serovar typhimurium. Both the extracts were screened for the presence of phytocompounds followed by the characterization using Attenuated Total Reflection (ATR) infrared spectroscopy and bioactivity finger print analysis. Anti-biofilm assays were determined to test the potential of both extracts to inhibit the biofilm formation, while Propidium Iodide (PI) uptake analysis revealed that cell membrane was damaged by the exposure of nutraceuticals for 1 h. This study has demonstrated that both nutraceuticals have anti-biofilm and antimicrobial activity perturbing the membrane integrity of food-borne S. typhimurium and could be used as curative remedy to control the food borne microbial infection. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

    PubMed

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

    2016-01-01

    Biofilms consist of surface-adhered bacterial communities encased in an extracellular matrix composed of DNA, exopolysaccharides, and proteins. Extracellular DNA (eDNA) has a structural role in the formation of biofilms, can bind and shield biofilms from aminoglycosides, and induces antimicrobial peptide resistance mechanisms. Here, we provide evidence that eDNA is responsible for the acidification of Pseudomonas aeruginosa planktonic cultures and biofilms. Further, we show that acidic pH and acidification via eDNA constitute a signal that is perceived by P. aeruginosa to induce the expression of genes regulated by the PhoPQ and PmrAB two-component regulatory systems. Planktonic P. aeruginosa cultured in exogenous 0.2% DNA or under acidic conditions demonstrates a 2- to 8-fold increase in aminoglycoside resistance. This resistance phenotype requires the aminoarabinose modification of lipid A and the production of spermidine on the bacterial outer membrane, which likely reduce the entry of aminoglycosides. Interestingly, the additions of the basic amino acid L-arginine and sodium bicarbonate neutralize the pH and restore P. aeruginosa susceptibility to aminoglycosides, even in the presence of eDNA. These data illustrate that the accumulation of eDNA in biofilms and infection sites can acidify the local environment and that acidic pH promotes the P. aeruginosa antibiotic resistance phenotype. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Effects of short-chain fatty acids on Actinomyces naeslundii biofilm formation.

    PubMed

    Yoneda, S; Kawarai, T; Narisawa, N; Tuna, E B; Sato, N; Tsugane, T; Saeki, Y; Ochiai, K; Senpuku, H

    2013-10-01

    Actinomyces naeslundii is an early colonizer and has important roles in the development of the oral biofilm. Short-chain fatty acids (SCFA) are secreted extracellularly as a product of metabolism by gram-negative anaerobes, e.g. Porphyromonas gingivalis and Fusobacterium nucleatum; and the SCFA may affect biofilm development with interaction between A. naeslundii and gram-negative bacteria. Our aim was to investigate the effects of SCFA on biofilm formation by A. naeslundii and to determine the mechanism. We used the biofilm formation assay in 96-well microtiter plates in tryptic soy broth without dextrose and with 0.25% sucrose using safranin stain of the biofilm monitoring 492 nm absorbance. To determine the mechanism by SCFA, the production of chaperones and stress-response proteins (GrpE and GroEL) in biofilm formation was examined using Western blot fluorescence activity with GrpE and GroEL antibodies. Adding butyric acid (6.25 mm) 0, 6 and 10 h after beginning culture significantly increased biofilm formation by A. naeslundii, and upregulation was observed at 16 h. Upregulation was also observed using appropriate concentrations of other SCFA. In the upregulated biofilm, production of GrpE and GroEL was higher where membrane-damaged or dead cells were also observed. The upregulated biofilm was significantly reduced by addition of anti-GroEL antibody. The data suggest biofilm formation by A. naeslundii was upregulated dependent on the production of stress proteins, and addition of SCFA increased membrane-damaged or dead cells. Production of GroEL may physically play an important role in biofilm development. 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

  9. Persister cells, the biofilm matrix and tolerance to metal cations in biofilm and planktonic Pseudomonas aeruginosa.

    PubMed

    Harrison, Joe J; Turner, Raymond J; Ceri, Howard

    2005-07-01

    In this study, we examined Pseudomonas aeruginosa ATCC 27853 biofilm and planktonic cell susceptibility to metal cations. The minimum inhibitory concentration (MIC), the minimum bactericidal concentration (MBC) required to eradicate 100% of the planktonic population (MBC 100), and the minimum biofilm eradication concentration (MBEC) were determined using the MBEC trade mark-high throughput assay. Six metals - Co(2+), Ni(2+), Cu(2+), Zn(2+), Al(3+) and Pb(2+)- were each tested at 2, 4, 6, 8, 10 and 27 h of exposure to biofilm and planktonic cultures grown in rich or minimal media. With 2 or 4 h of exposure, biofilms were approximately 2-25 times more tolerant to killing by metal cations than the corresponding planktonic cultures. However, by 27 h of exposure, biofilm and planktonic bacteria were eradicated at approximately the same concentration in every instance. Viable cell counts evaluated at 2 and 27 h of exposure revealed that at high concentrations, most of the metals assayed had killed greater than 99.9% of biofilm and planktonic cell populations. The surviving cells were propogated in vitro and gave rise to biofilm and planktonic cultures with normal sensitivity to metals. Further, retention of copper by the biofilm matrix was investigated using the chelator sodium diethlydithiocarbamate. Formation of visible brown metal-chelates in biofilms treated with Cu(2+) suggests that the biofilm matrix may coordinate and sequester metal cations from the aqueous surroundings. Overall, our data suggest that both metal sequestration in the biofilm matrix and the presence of a small population of 'persister' cells may be contributing factors in the time-dependent tolerance of both planktonic cells and biofilms to high concentrations of metal cations.

  10. Unexplored antifungal activity of linear battacin lipopeptides against planktonic and mature biofilms of C. albicans.

    PubMed

    De Zoysa, Gayan Heruka; Glossop, Hugh Douglas; Sarojini, Vijayalekshmi

    2018-02-25

    Novel antifungal agents are required against pathogenic fungi such as Candida albicans. We report the anticandidal activity of battacin lipopeptide antibiotics with previously unexplored antifungal activity. From amongst sixteen battacin lipopeptides tested against C. alibicans (SC5314) the 4-methyl hexanoyl conjugated trimeric lipopeptide 13 emerged as the lead candidate with a MIC of 6.25 μM and negligible haemolysis of mouse red blood cells. The potency of this lipopeptide was maintained under acidic conditions. Additionally, antifungal activity was further enhanced with amphotericin B at its non-haemolytic concentrations. Herein we have demonstrated for the first time that battacin lipopeptides prevent C. albicans biofilm colonisation as well as inhibit pre-formed biofilms of this fungal pathogen. XTT biofilm assays revealed that 13 prevented colonisation of C. albicans biofilms at its MIC (6.25 μM) and, at a higher concentration, eradicated 24 h (25 μM) and 48 h (62.5 μM) old preformed biofilms. In comparison, we found that amphotericin at much lower concentrations prevented biofilm colonisation (0.78 μM) and inhibited 24 h old preformed biofilms (6.25 μM), however was completely inactive against 48 h old preformed biofilms. Thus, lipopeptide 13 is more effective than amphotericin at eradicating more mature C. albicans biofilms. The membrane lytic mechanism of action of compound 13 was validated by a colorimetric assay using lipid vesicles mimicking fungal membranes in which compound 13 effected an immediate dark purple to red colour transition of suspended vesicles upon peptide interaction. In addition, TEM images of C. albicans cells exposed to 13 showed clearly disrupted cellular membranes. Interestingly, compound 13 increased the endogenous generation of reactive oxygen species (ROS) in a concentration dependent manner. In the presence of an antioxidant, ascorbic acid, ROS production was diminished yet antifungal activity

  11. Biofilms formed by the archaeon Haloferax volcanii exhibit cellular differentiation and social motility, and facilitate horizontal gene transfer.

    PubMed

    Chimileski, Scott; Franklin, Michael J; Papke, R Thane

    2014-08-14

    Archaea share a similar microbial lifestyle with bacteria, and not surprisingly then, also exist within matrix-enclosed communities known as biofilms. Advances in biofilm biology have been made over decades for model bacterial species, and include characterizations of social behaviors and cellular differentiation during biofilm development. Like bacteria, archaea impact ecological and biogeochemical systems. However, the biology of archaeal biofilms is only now being explored. Here, we investigated the development, composition and dynamics of biofilms formed by the haloarchaeon Haloferax volcanii DS2. Biofilms were cultured in static liquid and visualized with fluorescent cell membrane dyes and by engineering cells to express green fluorescent protein (GFP). Analysis by confocal scanning laser microscopy showed that H. volcanii cells formed microcolonies within 24 h, which developed into larger clusters by 48 h and matured into flake-like towers often greater than 100 μm in height after 7 days. To visualize the extracellular matrix, biofilms formed by GFP-expressing cells were stained with concanavalin A, DAPI, Congo red and thioflavin T. Stains colocalized with larger cellular structures and indicated that the extracellular matrix may contain a combination of polysaccharides, extracellular DNA and amyloid protein. Following a switch to biofilm growth conditions, a sub-population of cells differentiated into chains of long rods sometimes exceeding 25 μm in length, compared to their planktonic disk-shaped morphology. Time-lapse photography of static liquid biofilms also revealed wave-like social motility. Finally, we quantified gene exchange between biofilm cells, and found that it was equivalent to the mating frequency of a classic filter-based experimental method. The developmental processes, functional properties and dynamics of H. volcanii biofilms provide insight on how haloarchaeal species might persist, interact and exchange DNA in natural communities. H

  12. Sorption and Distribution of Copper in Unsaturated Pseudomonas putida CZ1 Biofilms as Determined by X-Ray Fluorescence Microscopy ▿

    PubMed Central

    Chen, Guangcun; Chen, Xincai; Yang, Yuanqiang; Hay, Anthony G.; Yu, Xiaohan; Chen, Yingxu

    2011-01-01

    The spatial and temporal distribution of metals in unsaturated Pseudomonas putida CZ1 biofilms was determined using synchrotron-based X-ray fluorescence microscopy (XRF). It was found that Fe, Mn, and Ca were mainly distributed near the air-biofilm interface of a biofilm grown on 40 mM citrate, while there were two Fe-, Mn-, and Ca-rich layers within a biofilm grown on 10 mM citrate. The sorption of copper by biofilm grown in medium containing 10 mM citrate was rapid, with copper being found throughout the biofilm after only 1 h of exposure. Copper initially colocalized with Fe and Mn element layers in the biofilm and then precipitated in a 40-μm-thick layer near the air-biofilm interface when exposed for 12 h. Cu K-edge X-ray absorption near edge structure (XANES) analysis revealed that Cu was primarily bound with citrate within the biofilm, and the precipitate formed in the biofilm exposed to copper for 12 h was most similar to copper phosphate. LIVE/DEAD staining revealed that cells at the biofilm-membrane interface were mostly alive even when the copper concentration reached 80.5 mg copper g−1 biomass. This suggests that the biofilm matrix provided significant protection for cells in this area. These results significantly improve our understanding of metal acquisition, transportation, and immobilization in unsaturated biofilm systems. PMID:21642411

  13. Concurrent microbial reduction of high concentrations of nitrate and perchlorate in an ion exchange membrane bioreactor.

    PubMed

    Fox, Shalom; Bruner, Tali; Oren, Yoram; Gilron, Jack; Ronen, Zeev

    2016-09-01

    We investigated effective simultaneous removal of high loads of nitrate and perchlorate from synthetic groundwater using an ion exchange membrane bioreactor (IEMB). The aim of this research was to characterize both transport aspects and biodegradation mechanisms involved in the treatment process of high loads of the two anions. Biodegradation process was proven to be efficient with over 99% efficiency of both perchlorate and nitrate, regardless of their load. The maximum biodegradation rates were 18.3 (mmol m(-2)  h(-1) ) and 5.5 (mmol m(-2)  h(-1) ) for nitrate and perchlorate, respectively. The presence of a biofilm on the bio-side of the membrane only slightly increased the nitrate and perchlorate transmembrane flux as compared to the measured flux during a Donnan dialysis experiment where there is no biodegradation of perchlorate and nitrate in the bio-compartment. The nitrate flux in presence of a biofilm was 18.3 (±1.9) (mmole m(-2)  h(-1) ), while without the biofilm, the flux was 16.9 (±1.5) (mmole m(-2)  h(-1) ) for the same feed inlet nitrate concentration of 4 mM. The perchlorate transmembrane flux increased similarly by an average of 5%. Samples of membrane biofilm and suspended bacteria from the bio-reactor were analyzed for diversity and abundance of the perchlorate and nitrate reducing bacteria. Klebsiella oxytoca, known as a glycerol fermenter, accounted for 70% of the suspended bacteria. In contrast, perchlorate and nitrate reducing bacteria predominated in the biofilm present on the membrane. These results are consistent with our proposed two stage biodegradation mechanism where glycerol is first fermented in the suspended phase of the bio-reactor and the fermentation products drive perchlorate and nitrate bio-reduction in the biofilm attached to the membrane. These results suggest that the niche exclusion of microbial populations in between the reactor and membrane is controlled by the fluxes of the electron donors and

  14. Biofilm Formation on Different Materials Used in Oral Rehabilitation.

    PubMed

    Souza, Júlio C M; Mota, Raquel R C; Sordi, Mariane B; Passoni, Bernardo B; Benfatti, Cesar A M; Magini, Ricardo S

    2016-01-01

    The aim of this study was to evaluate the density and the morphological aspects of biofilms adhered to different materials applied in oral rehabilitation supported by dental implants. Sixty samples were divided into four groups: feldspar-based porcelain, CoCr alloy, commercially pure titanium grade IV and yttria-stabilized zirconia. Human saliva was diluted into BHI supplemented with sucrose to grow biofilms for 24 or 48 h. After this period, biofilm was removed by 1% protease treatment and then analyzed by spectrophotometry (absorbance), colony forming unit method (CFU.cm-2) and field-emission guns scanning electron microscopy (FEG-SEM). The highest values of absorbance and CFU.cm-2 were recorded on biofilms grown on CoCr alloys when compared to the other test materials for 24 or 48 h. Also, FEG-SEM images showed a high biofilm density on CoCr. There were no significant differences in absorbance and CFU.cm-2 between biofilms grown on zirconia, porcelain and titanium (p<0.05). Microbiological assays associated with microscopic analyses detected a higher accumulation of oral biofilms on CoCr-based materials than that on titanium or zirconia that are used for prosthetic structures.

  15. Innovative sponge-based moving bed-osmotic membrane bioreactor hybrid system using a new class of draw solution for municipal wastewater treatment.

    PubMed

    Nguyen, Nguyen Cong; Chen, Shiao-Shing; Nguyen, Hau Thi; Ray, Saikat Sinha; Ngo, Huu Hao; Guo, Wenshan; Lin, Po-Hsun

    2016-03-15

    For the first time, an innovative concept of combining sponge-based moving bed (SMB) and an osmotic membrane bioreactor (OsMBR), known as the SMB-OsMBR hybrid system, were investigated using Triton X-114 surfactant coupled with MgCl2 salt as the draw solution. Compared to traditional activated sludge OsMBR, the SMB-OsMBR system was able to remove more nutrients due to the thick-biofilm layer on sponge carriers. Subsequently less membrane fouling was observed during the wastewater treatment process. A water flux of 11.38 L/(m(2h) and a negligible reverse salt flux were documented when deionized water served as the feed solution and a mixture of 1.5 M MgCl2 and 1.5 mM Triton X-114 was used as the draw solution. The SMB-OsMBR hybrid system indicated that a stable water flux of 10.5 L/(m(2h) and low salt accumulation were achieved in a 90-day operation. Moreover, the nutrient removal efficiency of the proposed system was close to 100%, confirming the effectiveness of simultaneous nitrification and denitrification in the biofilm layer on sponge carriers. The overall performance of the SMB-OsMBR hybrid system using MgCl2 coupled with Triton X-114 as the draw solution demonstrates its potential application in wastewater treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Sustained release of a novel anti-quorum-sensing agent against oral fungal biofilms.

    PubMed

    Feldman, Mark; Shenderovich, Julia; Al-Quntar, Abed Al Aziz; Friedman, Michael; Steinberg, Doron

    2015-04-01

    Thiazolidinedione-8 (S-8) has recently been identified as a potential anti-quorum-sensing/antibiofilm agent against bacteria and fungi. Based on these results, we investigated the possibility of incorporating S-8 in a sustained-release membrane (SRM) to increase its pharmaceutical potential against Candida albicans biofilm. We demonstrated that SRM containing S-8 inhibits fungal biofilm formation in a time-dependent manner for 72 h, due to prolonged release of S-8. Moreover, the SRM effectively delivered the agent in its active form to locations outside the membrane reservoir. In addition, eradication of mature biofilm by the SRM containing S-8 was also significant. Of note, S-8-containing SRM affected the characteristics of mature C. albicans biofilm, such as thickness, exopolysaccharide (EPS) production, and morphogenesis of fungal cells. The concept of using an antibiofilm agent with no antifungal activity incorporated into a sustained-release delivery system is new in medicine and dentistry. This concept of an SRM containing a quorum-sensing quencher with an antibiofilm effect could pave the way for combating oral fungal infectious diseases. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. Pentiptycene-Based Polyurethane with Enhanced Mechanical Properties and CO2-Plasticization Resistance for Thin Film Gas Separation Membranes.

    PubMed

    Pournaghshband Isfahani, Ali; Sadeghi, Morteza; Wakimoto, Kazuki; Shrestha, Binod Babu; Bagheri, Rouhollah; Sivaniah, Easan; Ghalei, Behnam

    2018-05-23

    The development of thin film composite (TFC) membranes offers an opportunity to achieve the permeability/selectivity requirements for optimum CO 2 separation performance. However, the durability and performance of thin film gas separation membranes are mostly challenged by weak mechanical properties and high CO 2 plasticization. Here, we designed new polyurethane (PU) structures with bulky aromatic chain extenders that afford preferred mechanical properties for ultra-thin-film formation. An improvement of about 1500% in Young's modulus and 600% in hardness was observed for pentiptycene-based PUs compared to the typical PU membranes. Single (CO 2 , H 2 , CH 4 , and N 2 ) and mixed (CO 2 /N 2 and CO 2 /CH 4 ) gas permeability tests were performed on the PU membranes. The resulting TFC membranes showed a high CO 2 permeance up to 1400 GPU (10 -6 cm 3 (STP) cm -2 s -1 cmHg -1 ) and the CO 2 /N 2 and CO 2 /H 2 selectivities of about 22 and 2.1, respectively. The enhanced mechanical properties of pentiptycene-based PUs result in high-performance thin membranes with the similar selectivity of the bulk polymer. The thin film membranes prepared from pentiptycene-based PUs also showed a twofold enhanced plasticization resistance compared to non-pentiptycene-containing PU membranes.

  18. Hyperosmotic Agents and Antibiotics Affect Dissolved Oxygen and pH Concentration Gradients in Staphylococcus aureus Biofilms

    PubMed Central

    Kiamco, Mia Mae; Atci, Erhan

    2017-01-01

    ABSTRACT Biofilms on wound surfaces are treated topically with hyperosmotic agents, such as medical-grade honey and cadexomer iodine; in some cases, these treatments are combined with antibiotics. Tissue repair requires oxygen, and a low pH is conducive to oxygen release from red blood cells and epithelialization. We investigated the variation of dissolved oxygen concentration and pH with biofilm depth and the variation in oxygen consumption rates when biofilms are challenged with medical-grade honey or cadexomer iodine combined with vancomycin or ciprofloxacin. Dissolved oxygen and pH depth profiles in Staphylococcus aureus biofilms were measured using microelectrodes. The presence of cadexomer iodine with vancomycin or ciprofloxacin on the surface of the biofilm permitted a measurable concentration of oxygen at greater biofilm depths (101.6 ± 27.3 μm, P = 0.02; and 155.5 ± 27.9 μm, P = 0.016, respectively) than in untreated controls (30.1 μm). Decreases in pH of ∼0.6 and ∼0.4 units were observed in biofilms challenged with medical-grade honey alone and combined with ciprofloxacin, respectively (P < 0.001 and 0.01, respectively); the number of bacteria recovered from biofilms was significantly reduced (1.26 log) by treatment with cadexomer iodine and ciprofloxacin (P = 0.002) compared to the untreated control. Combining cadexomer iodine and ciprofloxacin improved dissolved oxygen concentration and penetration depth into the biofilm, while medical-grade honey was associated with a lower pH; not all treatments established a bactericidal effect in the time frame used in the experiments. IMPORTANCE Reports about using hyperosmotic agents and antibiotics against wound biofilms focus mostly on killing bacteria, but the results of these treatments should additionally be considered in the context of how they affect physiologically important parameters, such as oxygen concentration and pH. We confirmed that the combination of a hyperosmotic agent and an antibiotic

  19. Hyperosmotic Agents and Antibiotics Affect Dissolved Oxygen and pH Concentration Gradients in Staphylococcus aureus Biofilms.

    PubMed

    Kiamco, Mia Mae; Atci, Erhan; Mohamed, Abdelrhman; Call, Douglas R; Beyenal, Haluk

    2017-03-15

    Biofilms on wound surfaces are treated topically with hyperosmotic agents, such as medical-grade honey and cadexomer iodine; in some cases, these treatments are combined with antibiotics. Tissue repair requires oxygen, and a low pH is conducive to oxygen release from red blood cells and epithelialization. We investigated the variation of dissolved oxygen concentration and pH with biofilm depth and the variation in oxygen consumption rates when biofilms are challenged with medical-grade honey or cadexomer iodine combined with vancomycin or ciprofloxacin. Dissolved oxygen and pH depth profiles in Staphylococcus aureus biofilms were measured using microelectrodes. The presence of cadexomer iodine with vancomycin or ciprofloxacin on the surface of the biofilm permitted a measurable concentration of oxygen at greater biofilm depths (101.6 ± 27.3 μm, P = 0.02; and 155.5 ± 27.9 μm, P = 0.016, respectively) than in untreated controls (30.1 μm). Decreases in pH of ∼0.6 and ∼0.4 units were observed in biofilms challenged with medical-grade honey alone and combined with ciprofloxacin, respectively ( P < 0.001 and 0.01, respectively); the number of bacteria recovered from biofilms was significantly reduced (1.26 log) by treatment with cadexomer iodine and ciprofloxacin ( P = 0.002) compared to the untreated control. Combining cadexomer iodine and ciprofloxacin improved dissolved oxygen concentration and penetration depth into the biofilm, while medical-grade honey was associated with a lower pH; not all treatments established a bactericidal effect in the time frame used in the experiments. IMPORTANCE Reports about using hyperosmotic agents and antibiotics against wound biofilms focus mostly on killing bacteria, but the results of these treatments should additionally be considered in the context of how they affect physiologically important parameters, such as oxygen concentration and pH. We confirmed that the combination of a hyperosmotic agent and an antibiotic

  20. Capillary-tube-based micro-plasma system for disinfecting dental biofilm.

    PubMed

    Huang, Wen-Ke; Weng, Chih-Chiang; Liao, Jiunn-Der; Wang, Yi-Cheng; Chuang, Shu-Fen

    2013-05-01

    A low-temperature low-energy capillary-tube-based argon micro-plasma system was applied to disinfect Streptococcus mutans-containing biofilm. The micro-plasma system uses a hollow inner electrode that is ignited by a radio-frequency power supply with a matching network. The energy content was analyzed using optical emission spectroscopy. The micro-plasma-induced effect on a biofilm cultured for 24 or 48 h with a working distance of ≈3 mm at low temperature was evaluated. The morphologies of the treated live/dead bacteria and the produced polysaccharides after micro-plasma treatment were examined. Scanning electron microscopy images and staining results show that most of the S. mutans on the treated biofilm were acutely damaged within a micro-plasma treatment time of 300 s. The number of living bacteria underneath the treated biofilm greatly decreased with treatment time. The proposed micro-plasma system can thus disinfect S. mutans on/in biofilms.

  1. Tannins Possessing Bacteriostatic Effect Impair Pseudomonas aeruginosa Adhesion and Biofilm Formation

    PubMed Central

    Trentin, Danielle S.; Silva, Denise B.; Amaral, Matheus W.; Zimmer, Karine R.; Silva, Márcia V.; Lopes, Norberto P.; Giordani, Raquel B.; Macedo, Alexandre J.

    2013-01-01

    Plants produce many compounds that are biologically active, either as part of their normal program of growth and development or in response to pathogen attack or stress. Traditionally, Anadenanthera colubrina, Commiphora leptophloeos and Myracrodruon urundeuva have been used by communities in the Brazilian Caatinga to treat several infectious diseases. The ability to impair bacterial adhesion represents an ideal strategy to combat bacterial pathogenesis, because of its importance in the early stages of the infectious process; thus, the search for anti-adherent compounds in plants is a very promising alternative. This study investigated the ability of stem-bark extracts from these three species to control the growth and prevent biofilm formation of Pseudomonas aeruginosa, an important opportunistic pathogen that adheres to surfaces and forms protective biofilms. A kinetic study (0–72 h) demonstrated that the growth of extract-treated bacteria was inhibited up to 9 h after incubation, suggesting a bacteriostatic activity. Transmission electron microscopy and fluorescence microscopy showed both viable and nonviable cells, indicating bacterial membrane damage; crystal violet assay and scanning electron microscopy demonstrated that treatment strongly inhibited biofilm formation during 6 and 24 h and that matrix production remained impaired even after growth was restored, at 24 and 48 h of incubation. Herein, we propose that the identified (condensed and hydrolyzable) tannins are able to inhibit biofilm formation via bacteriostatic properties, damaging the bacterial membrane and hindering matrix production. Our findings demonstrate the importance of this abundant class of Natural Products in higher plants against one of the most challenging issues in the hospital setting: biofilm resilience. PMID:23776646

  2. Investigating biofilm structure using x-ray microtomography and gratings-based phase contrast

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Miller, Erin A.; Xiao, Xianghui; Miller, Micah D.

    2012-10-17

    Direct examination of natural and engineered environments has revealed that the majority of microorganisms in these systems live in structured communities termed biofilms. To gain a better understanding for how biofilms function and interact with their local environment, fundamental capabilities for enhanced visualization, compositional analysis, and functional characterization of biofilms are needed. For pore-scale and community-scale analysis (100’s of nm to 10’s of microns), a variety of surface tools are available. However, understanding biofilm structure in complex three-dimensional (3-D) environments is considerably more difficult. X-ray microtomography can reveal a biofilm’s internal structure, but the obtaining sufficient contrast to image low-Zmore » biological material against a higher-Z substrate makes detecting biofilms difficult. Here we present results imaging Shewanella oneidensis biofilms on a Hollow-fiber Membrane Biofilm Reactor (HfMBR), using the x-ray microtomography system at sector 2-BM of the Advanced Photon Source (APS), at energies ranging from 13-15.4 keV and pixel sizes of 0.7 and 1.3 μm/pixel. We examine the use of osmium (Os) as a contrast agent to enhance biofilm visibility and demonstrate that staining improves imaging of hydrated biofilms. We also present results using a Talbot interferometer to provide phase and scatter contrast information in addition to absorption. Talbot interferometry allows imaging of unstained hydrated biofilms with phase contrast, while absorption contrast primarily highlights edges and scatter contrast provides little information. However, the gratings used here limit the spatial resolution to no finer than 2 μm, which hinders the ability to detect small features. Future studies at higher resolution or higher Talbot order for greater sensitivity to density variations may improve imaging.« less

  3. The application of nitric oxide to control biofouling of membrane bioreactors

    PubMed Central

    Luo, Jinxue; Zhang, Jinsong; Barnes, Robert J; Tan, Xiaohui; McDougald, Diane; Fane, Anthony G; Zhuang, Guoqiang; Kjelleberg, Staffan; Cohen, Yehuda; Rice, Scott A

    2015-01-01

    A novel strategy to control membrane bioreactor (MBR) biofouling using the nitric oxide (NO) donor compound PROLI NONOate was examined. When the biofilm was pre-established on membranes at transmembrane pressure (TMP) of 88–90 kPa, backwashing of the membrane module with 80 μM PROLI NONOate for 45 min once daily for 37 days reduced the fouling resistance (Rf) by 56%. Similarly, a daily, 1 h exposure of the membrane to 80 μM PROLI NONOate from the commencement of MBR operation for 85 days resulted in reduction of the TMP and Rf by 32.3% and 28.2%. The microbial community in the control MBR was observed to change from days 71 to 85, which correlates with the rapid TMP increase. Interestingly, NO-treated biofilms at 85 days had a higher similarity with the control biofilms at 71 days relative to the control biofilms at 85 days, indicating that the NO treatment delayed the development of biofilm bacterial community. Despite this difference, sequence analysis indicated that NO treatment did not result in a significant shift in the dominant fouling species. Confocal microscopy revealed that the biomass of biopolymers and microorganisms in biofilms were all reduced on the PROLI NONOate-treated membranes, where there were reductions of 37.7% for proteins and 66.7% for microbial cells, which correlates with the reduction in TMP. These results suggest that NO treatment could be a promising strategy to control biofouling in MBRs. PMID:25752591

  4. Variation in Biofilm Stability with Decreasing pH Affects Porous Medium Hydraulic Properties

    NASA Astrophysics Data System (ADS)

    Kirk, M. F.; Santillan, E. F.; McGrath, L. K.; Altman, S. J.

    2010-12-01

    Changes to microbial communities caused by subsurface CO2 injection may have many consequences, including possible impacts to CO2 transport. We used column experiments to examine how decreasing pH, a geochemical change associated with CO2 injection, will affect biofilm stability and ultimately the hydraulic properties of porous media. Columns consisted of 1 mm2 square capillary tubes filled with 105-150 µm diameter glass beads. Artificial groundwater medium containing 1 mM glucose was pumped through the columns at a rate of 0.01 mL/min (q = 14.4 m/day; Re = 0.03). Columns were inoculated with 3 × 10^8 CFU (avg.) of Pseudomonas fluorescens, a model biofilm former, transformed with a green fluorescent protein. Biomass distribution and transport was examined using scanning laser confocal microscopy and effluent plating. Variation in the bulk hydraulic properties of the columns was measured using manometers. In an initial experiment, biofilm growth was allowed to occur for seven days in medium with pH 7.3. Within this period, cells uniformly coated bead surfaces, effluent cell numbers stabilized at 1 × 10^9 CFU/mL, and hydraulic conductivity (K) decreased 77%. Next, medium with pH 4 was introduced. As a result, biomass within the reactor redistributed from bead surfaces to pores, effluent cell numbers decreased to 3 × 10^5 CFU/mL, and K decreased even further (>94% reduction). This decreased K was maintained until the experiment was terminated, seven days after introducing low pH medium. These results suggest that changes in biomass distribution as a result of decreased pH may initially limit transport of solubility-trapped CO2 following CO2 injection. Experiments in progress and planned will test this result in more detail and over longer periods of time. This material is based upon work supported as part of the Center for Frontiers of Subsurface Energy Security, an Energy Frontier Research Center funded by the U.S. Department of Energy, Office of Science, Office

  5. Development of an energy-saving anaerobic hybrid membrane bioreactors for 2-chlorophenol-contained wastewater treatment.

    PubMed

    Wang, Yun-Kun; Pan, Xin-Rong; Sheng, Guo-Ping; Li, Wen-Wei; Shi, Bing-Jing; Yu, Han-Qing

    2015-12-01

    A novel energy-saving anaerobic hybrid membrane bioreactor (AnHMBR) with mesh filter, which takes advantage of anaerobic membrane bioreactor and fixed-bed biofilm reactor, is developed for low-strength 2-chlorophenol (2-CP)-contained wastewater treatment. In this system, the anaerobic membrane bioreactor is stuffed with granular activated carbon to construct an anaerobic hybrid fixed-bed biofilm membrane bioreactor. The effluent turbidity from the AnHMBR system was low during most of the operation period, and the chemical oxygen demand and 2-CP removal efficiencies averaged 82.3% and 92.6%, respectively. Furthermore, a low membrane fouling rate was achieved during the operation. During the AnHMBR operation, the only energy consumption was for feed pump. And a low energy demand of 0.0045-0.0063kWhm(-3) was estimated under the current operation conditions. All these results demonstrated that this novel AnHMBR is a sustainable technology for treating 2-CP-contained wastewater. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Syntrophic growth of Desulfovibrio alaskensis requires genes for H2 and formate metabolism as well as those for flagellum and biofilm formation.

    PubMed

    Krumholz, Lee R; Bradstock, Peter; Sheik, Cody S; Diao, Yiwei; Gazioglu, Ozcan; Gorby, Yuri; McInerney, Michael J

    2015-04-01

    In anaerobic environments, mutually beneficial metabolic interactions between microorganisms (syntrophy) are essential for oxidation of organic matter to carbon dioxide and methane. Syntrophic interactions typically involve a microorganism degrading an organic compound to primary fermentation by-products and sources of electrons (i.e., formate, hydrogen, or nanowires) and a partner producing methane or respiring the electrons via alternative electron accepting processes. Using a transposon gene mutant library of the sulfate-reducing Desulfovibrio alaskensis G20, we screened for mutants incapable of serving as the electron-accepting partner of the butyrate-oxidizing bacterium, Syntrophomonas wolfei. A total of 17 gene mutants of D. alaskensis were identified as incapable of serving as the electron-accepting partner. The genes identified predominantly fell into three categories: membrane surface assembly, flagellum-pilus synthesis, and energy metabolism. Among these genes required to serve as the electron-accepting partner, the glycosyltransferase, pilus assembly protein (tadC), and flagellar biosynthesis protein showed reduced biofilm formation, suggesting that each of these components is involved in cell-to-cell interactions. Energy metabolism genes encoded proteins primarily involved in H2 uptake and electron cycling, including a rhodanese-containing complex that is phylogenetically conserved among sulfate-reducing Deltaproteobacteria. Utilizing an mRNA sequencing approach, analysis of transcript abundance in wild-type axenic and cocultures confirmed that genes identified as important for serving as the electron-accepting partner were more highly expressed under syntrophic conditions. The results imply that sulfate-reducing microorganisms require flagellar and outer membrane components to effectively couple to their syntrophic partners; furthermore, H2 metabolism is essential for syntrophic growth of D. alaskensis G20. Copyright © 2015, American Society for

  7. Syntrophic Growth of Desulfovibrio alaskensis Requires Genes for H2 and Formate Metabolism as Well as Those for Flagellum and Biofilm Formation

    PubMed Central

    Bradstock, Peter; Sheik, Cody S.; Diao, Yiwei; Gazioglu, Ozcan; Gorby, Yuri; McInerney, Michael J.

    2015-01-01

    In anaerobic environments, mutually beneficial metabolic interactions between microorganisms (syntrophy) are essential for oxidation of organic matter to carbon dioxide and methane. Syntrophic interactions typically involve a microorganism degrading an organic compound to primary fermentation by-products and sources of electrons (i.e., formate, hydrogen, or nanowires) and a partner producing methane or respiring the electrons via alternative electron accepting processes. Using a transposon gene mutant library of the sulfate-reducing Desulfovibrio alaskensis G20, we screened for mutants incapable of serving as the electron-accepting partner of the butyrate-oxidizing bacterium, Syntrophomonas wolfei. A total of 17 gene mutants of D. alaskensis were identified as incapable of serving as the electron-accepting partner. The genes identified predominantly fell into three categories: membrane surface assembly, flagellum-pilus synthesis, and energy metabolism. Among these genes required to serve as the electron-accepting partner, the glycosyltransferase, pilus assembly protein (tadC), and flagellar biosynthesis protein showed reduced biofilm formation, suggesting that each of these components is involved in cell-to-cell interactions. Energy metabolism genes encoded proteins primarily involved in H2 uptake and electron cycling, including a rhodanese-containing complex that is phylogenetically conserved among sulfate-reducing Deltaproteobacteria. Utilizing an mRNA sequencing approach, analysis of transcript abundance in wild-type axenic and cocultures confirmed that genes identified as important for serving as the electron-accepting partner were more highly expressed under syntrophic conditions. The results imply that sulfate-reducing microorganisms require flagellar and outer membrane components to effectively couple to their syntrophic partners; furthermore, H2 metabolism is essential for syntrophic growth of D. alaskensis G20. PMID:25616787

  8. Impact of membrane characteristics on the performance and cycling of the Br-2-H-2 redox flow cell

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tucker, MC; Cho, KT; Spingler, FB

    2015-06-15

    The Br-2/H-2 redox flow cell shows promise as a high-power, low-cost energy storage device. In this paper, the effect of various aspects of material selection and processing of proton exchange membranes on the operation of the Br-2/H-2 redox flow cell is determined. Membrane properties have a significant impact on the performance and efficiency of the system. In particular, there is a tradeoff between conductivity and crossover, where conductivity limits system efficiency at high current density and crossover limits efficiency at low current density. The impact of thickness, pretreatment procedure, swelling state during cell assembly, equivalent weight, membrane reinforcement, and additionmore » of a microporous separator layer on this tradeoff is assessed. NR212 (50 mu m) pretreated by soaking in 70 degrees C water is found to be optimal for the studied operating conditions. For this case, an energy efficiency of greater than 75% is achieved for current density up to 400 mA cm(-2), with a maximum obtainable energy efficiency of 88%. A cell with this membrane was cycled continuously for 3164 h. Membrane transport properties, including conductivity and bromine and water crossover, were found to decrease moderately upon cycling but remained higher than those for the as-received membrane. (C) 2015 Elsevier B.V. All rights reserved.« less

  9. A Multispecies Fungal Biofilm Approach to Enhance the Celluloyltic Efficiency of Membrane Reactors for Consolidated Bioprocessing of Plant Biomass

    PubMed Central

    Xiros, Charilaos; Studer, Michael H.

    2017-01-01

    The constraints and advantages in cellulolytic enzymes production by fungal biofilms for a consolidated bioconversion process were investigated during this study. The biofilm cultivations were carried out in reactors designed for consolidated bioprocessing Multispecies Biofilm Membrane reactors, (MBM) where an aerobic fungal biofilm produces the lignocellulolytic enzymes while a fermenting microorganism forms the fermentation product at anaerobic conditions. It was shown that although mycelial growth was limited in the MBM reactors compared to submerged cultivations, the secretion of cellulolytic enzymes per cell dry weight was higher. When Trichoderma reesei was used as the sole enzyme producer, cellobiose accumulated in the liquid medium as the result of the deficiency of β-glucosidase in the fungal secretome. To enhance β-glucosidase activity, T. reesei was co-cultivated with A. phoenicis which is a β-glucosidase overproducer. The two fungi formed a multispecies biofilm which produced a balanced cellulolytic cocktail for the saccharification of plant biomass. The mixed biofilm reached a 2.5 fold increase in β-glucosidase production, compared to the single T. reesei biofilm. The enzymatic systems of single and mixed biofilms were evaluated regarding their efficiency on cellulosic substrates degradation. Washed solids from steam pretreated beechwood, as well as microcrystalline cellulose were used as the substrates. The enzymatic system of the multispecies biofilm released four times more glucose than the enzymatic system of T. reesei alone from both substrates and hydrolyzed 78 and 60% of the cellulose content of washed solids from beechwood and microcrystalline cellulose, respectively. PMID:29067006

  10. Polyamines cause plasma membrane depolarization, activate Ca2+-, and modulate H+-ATPase pump activity in pea roots.

    PubMed

    Pottosin, Igor; Velarde-Buendía, Ana María; Bose, Jayakumar; Fuglsang, Anja T; Shabala, Sergey

    2014-06-01

    Polyamines regulate a variety of cation and K(+) channels, but their potential effects on cation-transporting ATPases are underexplored. In this work, noninvasive microelectrode ion flux estimation and conventional microelectrode techniques were applied to study the effects of polyamines on Ca(2+) and H(+) transport and membrane potential in pea roots. Externally applied spermine or putrescine (1mM) equally activated eosin yellow (EY)-sensitive Ca(2+) pumping across the root epidermis and caused net H(+) influx or efflux. Proton influx induced by spermine was suppressed by EY, supporting the mechanism in which Ca(2+) pump imports 2 H(+) per each exported Ca(2+). Suppression of the Ca(2+) pump by EY diminished putrescine-induced net H(+) efflux instead of increasing it. Thus, activities of Ca(2+) and H(+) pumps were coupled, likely due to the H(+)-pump inhibition by intracellular Ca(2+). Additionally, spermine but not putrescine caused a direct inhibition of H(+) pumping in isolated plasma membrane vesicles. Spermine, spermidine, and putrescine (1mM) induced membrane depolarization by 70, 50, and 35 mV, respectively. Spermine-induced depolarization was abolished by cation transport blocker Gd(3+), was insensitive to anion channels' blocker niflumate, and was dependent on external Ca(2+). Further analysis showed that uptake of polyamines but not polyamine-induced cationic (K(+)+Ca(2+)+H(+)) fluxes were a main cause of membrane depolarization. Polyamine increase is a common component of plant stress responses. Activation of Ca(2+) efflux by polyamines and contrasting effects of polyamines on net H(+) fluxes and membrane potential can contribute to Ca(2+) signalling and modulate a variety of transport processes across the plasma membrane under stress. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  11. Biofilm Matrix Proteins.

    PubMed

    Fong, Jiunn N C; Yildiz, Fitnat H

    2015-04-01

    Proteinaceous components of the biofilm matrix include secreted extracellular proteins, cell surface adhesins, and protein subunits of cell appendages such as flagella and pili. Biofilm matrix proteins play diverse roles in biofilm formation and dissolution. They are involved in attaching cells to surfaces, stabilizing the biofilm matrix via interactions with exopolysaccharide and nucleic acid components, developing three-dimensional biofilm architectures, and dissolving biofilm matrix via enzymatic degradation of polysaccharides, proteins, and nucleic acids. In this article, we will review functions of matrix proteins in a selected set of microorganisms, studies of the matrix proteomes of Vibrio cholerae and Pseudomonas aeruginosa, and roles of outer membrane vesicles and of nucleoid-binding proteins in biofilm formation.

  12. Antimicrobial activity of hydroxyl radicals generated by hydrogen peroxide photolysis against Streptococcus mutans biofilm.

    PubMed

    Nakamura, Keisuke; Shirato, Midori; Kanno, Taro; Örtengren, Ulf; Lingström, Peter; Niwano, Yoshimi

    2016-10-01

    Prevention of dental caries with maximum conservation of intact tooth substance remains a challenge in dentistry. The present study aimed to evaluate the antimicrobial effect of H2O2 photolysis on Streptococcus mutans biofilm, which may be a novel antimicrobial chemotherapy for treating caries. S. mutans biofilm was grown on disk-shaped hydroxyapatite specimens. After 1-24 h of incubation, growth was assessed by confocal laser scanning microscopy and viable bacterial counting. Resistance to antibiotics (amoxicillin and erythromycin) was evaluated by comparing bactericidal effects on the biofilm with those on planktonic bacteria. To evaluate the effect of the antimicrobial technique, the biofilm was immersed in 3% H2O2 and was irradiated with an LED at 365 nm for 1 min. Viable bacterial counts in the biofilm were determined by colony counting. The thickness and surface coverage of S. mutans biofilm increased with time, whereas viable bacterial counts plateaued after 6 h. When 12- and 24-h-old biofilms were treated with the minimum concentration of antibiotics that killed viable planktonic bacteria with 3 log reduction, their viable counts were not significantly decreased, suggesting the biofilm acquired antibiotic resistance by increasing its thickness. By contrast, hydroxyl radicals generated by photolysis of 3% H2O2 effectively killed S. mutans in 24-h-old biofilm, with greater than 5 log reduction. The technique based on H2O2 photolysis is a potentially powerful adjunctive antimicrobial chemotherapy for caries treatment. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  13. Nitric Oxide Treatment for the Control of Reverse Osmosis Membrane Biofouling

    PubMed Central

    Barnes, Robert J.; Low, Jiun Hui; Bandi, Ratnaharika R.; Tay, Martin; Chua, Felicia; Aung, Theingi; Fane, Anthony G.; Kjelleberg, Staffan

    2015-01-01

    Biofouling remains a key challenge for membrane-based water treatment systems. This study investigated the dispersal potential of the nitric oxide (NO) donor compound, PROLI NONOate, on single- and mixed-species biofilms formed by bacteria isolated from industrial membrane bioreactor and reverse osmosis (RO) membranes. The potential of PROLI NONOate to control RO membrane biofouling was also examined. Confocal microscopy revealed that PROLI NONOate exposure induced biofilm dispersal in all but two of the bacteria tested and successfully dispersed mixed-species biofilms. The addition of 40 μM PROLI NONOate at 24-h intervals to a laboratory-scale RO system led to a 92% reduction in the rate of biofouling (pressure rise over a given period) by a bacterial community cultured from an industrial RO membrane. Confocal microscopy and extracellular polymeric substances (EPS) extraction revealed that PROLI NONOate treatment led to a 48% reduction in polysaccharides, a 66% reduction in proteins, and a 29% reduction in microbial cells compared to the untreated control. A reduction in biofilm surface coverage (59% compared to 98%, treated compared to control) and average thickness (20 μm compared to 26 μm, treated compared to control) was also observed. The addition of PROLI NONOate led to a 22% increase in the time required for the RO module to reach its maximum transmembrane pressure (TMP), further indicating that NO treatment delayed fouling. Pyrosequencing analysis revealed that the NO treatment did not significantly alter the microbial community composition of the membrane biofilm. These results present strong evidence for the application of PROLI NONOate for prevention of RO biofouling. PMID:25636842

  14. Sequential effect of phages and cold nitrogen plasma against Escherichia coli O157:H7 biofilms on different vegetables.

    PubMed

    Cui, Haiying; Bai, Mei; Yuan, Lu; Surendhiran, Duraiarasan; Lin, Lin

    2018-03-02

    Escherichia coli O157:H7 (E. coli O157:H7) is one of the most common pathogens in fresh vegetables and fruits, and most of the diseases produced by E. coli O157:H7 are associated with biofilms. Cold nitrogen plasma (CNP) is a cold sterilization technique which has no residue. However to completely eliminate the biofilm on the surface of vegetables the processing power and time of CNP have to be enhanced, which will impact on the quality of fruits and vegetables. Thus the sequential treatment of CNP and phage techniques was engineered in this study. Compared to treatment performed separately, sequential treatment not only had more mild treatment conditions as 400W CNP treatment for 2min and 5% phage treatment for 30min, but also exhibited more remarkable effect on eradicating E. coli O157:H7 biofilms in vitro and on vegetables. The population of E. coli O157:H7 was approximately reduced by 2logCFU/cm 2 after individual treatment of 5% phages for 30min or 500W CNP for 3min. While the sequential treatment of CNP (400W, 2min) and phages (5%, 30min) reduced the E. coli O157:H7 viable count in biofilm by 5.71logCFU/cm 2 . Therefore, the sequential treatment holds a great promise to improve the current treatment systems of bacterial contamination on different vegetable surfaces. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Deactivation of Pt/VC proton exchange membrane fuel cell cathodes by SO2, H2S and COS

    NASA Astrophysics Data System (ADS)

    Gould, Benjamin D.; Baturina, Olga A.; Swider-Lyons, Karen E.

    Sulfur contaminants in air pose a threat to the successful operation of proton exchange membrane fuel cells (PEMFCs) via poisoning of the Pt-based cathodes. The deactivation behavior of commercial Pt on Vulcan carbon (Pt/VC) membrane electrode assemblies (MEAs) is determined when exposed to 1 ppm (dry) of SO 2, H 2S, or COS in air for 3, 12, and 24 h while held at a constant potential of 0.6 V. All the three sulfur compounds cause the same deactivation behavior in the fuel cell cathodes, and the polarization curves of the poisoned MEAs have the same decrease in performance. Sulfur coverages after multiple exposure times (3, 12, and 24 h) are determined by cyclic voltammetry (CV). As the exposure time to sulfur contaminants increases from 12 to 24 h, the sulfur coverage of the platinum saturates at 0.45. The sulfur is removed from the cathodes and their activity is partially restored both by cyclic voltammetry, as shown by others, and by successive polarization curves. Complete recovery of fuel cell performance is not achieved with either technique, suggesting that sulfur species permanently affect the surface of the catalyst.

  16. Efficacy of a surfactant-based wound dressing on biofilm control.

    PubMed

    Percival, Steven L; Mayer, Dieter; Salisbury, Anne-Marie

    2017-09-01

    The aim of this study was to evaluate the efficacy of both a nonantimicrobial and antimicrobial (1% silver sulfadiazine-SSD) surfactant-based wound dressing in the control of Pseudomonas aeruginosa, Enterococcus sp, Staphylococcus epidermidis, Staphylococcus aureus, and methicillin-resistant S. aureus (MRSA) biofilms. Anti-biofilm efficacy was evaluated in numerous adapted American Standards for Testing and Materials (ASTM) standard biofilm models and other bespoke biofilm models. The ASTM standard models employed included the Minimum biofilm eradication concentration (MBEC) biofilm model (ASTM E2799) and the Centers for Disease Control (CDC) biofilm reactor model (ASTM 2871). Such bespoke biofilm models included the filter biofilm model and the chamberslide biofilm model. Results showed complete kill of microorganisms within a biofilm using the antimicrobial surfactant-based wound dressing. Interestingly, the nonantimicrobial surfactant-based dressing could disrupt existing biofilms by causing biofilm detachment. Prior to biofilm detachment, we demonstrated, using confocal laser scanning microscopy (CLSM), the dispersive effect of the nonantimicrobial surfactant-based wound dressing on the biofilm within 10 minutes of treatment. Furthermore, the non-antimicrobial surfactant-based wound dressing caused an increase in microbial flocculation/aggregation, important for microbial concentration. In conclusion, this nonantimicrobial surfactant-based wound dressing leads to the effective detachment and dispersion of in vitro biofilms. The use of surfactant-based wound dressings in a clinical setting may help to disrupt existing biofilm from wound tissue and may increase the action of antimicrobial treatment. © 2017 by the Wound Healing Society.

  17. Modeling of simultaneous anaerobic methane and ammonium oxidation in a membrane biofilm reactor.

    PubMed

    Chen, Xueming; Guo, Jianhua; Shi, Ying; Hu, Shihu; Yuan, Zhiguo; Ni, Bing-Jie

    2014-08-19

    Nitrogen removal by using the synergy of denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (Anammox) microorganisms in a membrane biofilm reactor (MBfR) has previously been demonstrated experimentally. In this work, a mathematical model is developed to describe the simultaneous anaerobic methane and ammonium oxidation by DAMO and Anammox microorganisms in an MBfR for the first time. In this model, DAMO archaea convert nitrate, both externally fed and/or produced by Anammox, to nitrite, with methane as the electron donor. Anammox and DAMO bacteria jointly remove the nitrite fed/produced, with ammonium and methane as the electron donor, respectively. The model is successfully calibrated and validated using the long-term (over 400 days) dynamic experimental data from the MBfR, as well as two independent batch tests at different operational stages of the MBfR. The model satisfactorily describes the methane oxidation and nitrogen conversion data from the system. Modeling results show the concentration gradients of methane and nitrogen would cause stratification of the biofilm, where Anammox bacteria mainly grow in the biofilm layer close to the bulk liquid and DAMO organisms attach close to the membrane surface. The low surface methane loadings result in a low fraction of DAMO microorganisms, but the high surface methane loadings would lead to overgrowth of DAMO bacteria, which would compete with Anammox for nitrite and decrease the fraction of Anammox bacteria. The results suggest an optimal methane supply under the given condition should be applied not only to benefit the nitrogen removal but also to avoid potential methane emissions.

  18. Effects of environmental parameters on the dual-species biofilms formed by Escherichia coli O157:H7 and Ralstonia insidiosa, a strong biofilm producer isolated from a fresh-cut produce processing plant.

    PubMed

    Liu, Nancy T; Nou, Xiangwu; Bauchan, Gary R; Murphy, Charles; Lefcourt, Alan M; Shelton, Daniel R; Lo, Y Martin

    2015-01-01

    Biofilm-forming bacteria resident to food processing facilities are a food safety concern due to the potential of biofilms to harbor foodborne bacterial pathogens. When cultured together, Ralstonia insidiosa, a strong biofilm former frequently isolated from produce processing environments, has been shown to promote the incorporation of Escherichia coli O157:H7 into dual-species biofilms. In this study, interactions between E. coli O157:H7 and R. insidiosa were examined under different incubating conditions. Under static culture conditions, the incorporation of E. coli O157:H7 into biofilms with R. insidiosa was not significantly affected by either low incubating temperature (10°C) or by limited nutrient availability. Greater enhancement of E. coli O157:H7 incorporation in dual-species biofilms was observed by using a continuous culture system with limited nutrient availability. Under the continuous culture conditions used in this study, E coli O157:H7 cells showed a strong tendency of colocalizing with R. insidiosa on a glass surface at the early stage of biofilm formation. As the biofilms matured, E coli O157:H7 cells were mostly found at the bottom layer of the dual-species biofilms, suggesting an effective protection by R. insidiosa in the mature biofilms.

  19. Phenol removal performance and microbial community shift during pH shock in a moving bed biofilm reactor (MBBR).

    PubMed

    Zhou, Hao; Wang, Guochen; Wu, Minghuo; Xu, Weiping; Zhang, Xuwang; Liu, Lifen

    2018-06-05

    A moving bed biofilm reactor (MBBR) effectively removes pollutants and even runs under extreme conditions. However, the pH shock resistance of a biofilm in MBBRs has been rarely reported. In this study, simulated phenol wastewater with acidic shock (pH 7.5-3.0) was used. In the pH shock phase, the phenol and COD removal efficiencies initially decreased and gradually increased to more than 90%. Microscopic studies showed that the superficial biofilm was mainly composed of fungi (yeasts) in the acidic pH shock phase. The microbial community composition in the acidic pH shock phase was significantly different from those in other phases. Firmicutes and Ascomycota were the dominant bacterial and fungal phyla in this stage, respectively. 16S rRNA gene-based functional annotation indicated that functional profiles related to aromatic compound degradation existed in all of the stages. Therefore, MBBRs show potential for the treatment of phenolic wastewater exposed to pH shock. Copyright © 2018 Elsevier B.V. All rights reserved.

  20. Chitosan-phosphotungstic acid complex as membranes for low temperature H2-O2 fuel cell

    NASA Astrophysics Data System (ADS)

    Santamaria, M.; Pecoraro, C. M.; Di Quarto, F.; Bocchetta, P.

    2015-02-01

    Free-standing Chitosan/phosphotungstic acid polyelectrolyte membranes were prepared by an easy and fast in-situ ionotropic gelation process performed at room temperature. Scanning electron microscopy was employed to study their morphological features and their thickness as a function of the chitosan concentration. The membrane was tested as proton conductor in low temperature H2-O2 fuel cell allowing to get peak power densities up to 350 mW cm-2. Electrochemical impedance measurements allowed to estimate a polyelectrolyte conductivity of 18 mS cm-1.

  1. Candida tropicalis biofilm and human epithelium invasion is highly influenced by environmental pH.

    PubMed

    Ferreira, Carina; Gonçalves, Bruna; Vilas Boas, Diana; Oliveira, Hugo; Henriques, Mariana; Azeredo, Joana; Silva, Sónia

    2016-11-01

    The main goal of this study was to investigate the role of pH on Candida tropicalis virulence determinants, namely the ability to form biofilms and to colonize/invade reconstituted human vaginal epithelia. Biofilm formation was evaluated by enumeration of cultivable cells, total biomass quantification and structural analysis by scanning electron microscopy and confocal laser scanning microscopy. Candida tropicalis human vaginal epithelium colonization and invasiveness were examined qualitatively by epifluorescence microscopy and quantitatively by a novel quantitative real-time PCR protocol for Candida quantification in tissues. The results revealed that environmental pH influences C. tropicalis biofilm formation as well as the colonization and potential to invade human epithelium with intensification at neutral and alkaline conditions compared to acidic conditions. For the first time, we have demonstrated that C. tropicalis biofilm formation and invasion is highly influenced by environmental pH. © Crown copyright 2016.

  2. A feasibility study on the bioconversion of CO2 and H2 to biomethane by gas sparging through polymeric membranes.

    PubMed

    Díaz, I; Pérez, C; Alfaro, N; Fdz-Polanco, F

    2015-06-01

    In this study, the potential of a pilot hollow-fiber membrane bioreactor for the conversion of H2 and CO2 to CH4 was evaluated. The system transformed 95% of H2 and CO2 fed at a maximum loading rate of 40.2 [Formula: see text] and produced 0.22m(3) of CH4 per m(3) of H2 fed at thermophilic conditions. H2 mass transfer to the liquid phase was identified as the limiting step for the conversion, and kLa values of 430h(-1) were reached in the bioreactor by sparging gas through the membrane module. A simulation showed that the bioreactor could upgrade biogas at a rate of 25m(3)/mR(3)d, increasing the CH4 concentration from 60 to 95%v. This proof-of-concept study verified that gas sparging through a membrane module can efficiently transfer H2 from gas to liquid phase and that the conversion of H2 and CO2 to biomethane is feasible on a pilot scale at noteworthy load rates. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. S-aryl-L-cysteine sulphoxides and related organosulphur compounds alter oral biofilm development and AI-2-based cell-cell communication.

    PubMed

    Kasper, S H; Samarian, D; Jadhav, A P; Rickard, A H; Musah, R A; Cady, N C

    2014-11-01

    To design and synthesize a library of structurally related, small molecules related to homologues of compounds produced by the plant Petiveria alliacea and determine their ability to interfere with AI-2 cell-cell communication and biofilm formation by oral bacteria. Many human diseases are associated with persistent bacterial biofilms. Oral biofilms (dental plaque) are problematic as they are often associated with tooth decay, periodontal disease and systemic disorders such as heart disease and diabetes. Using a microplate-based approach, a bio-inspired small molecule library was screened for anti-biofilm activity against the oral species Streptococcus mutans UA159, Streptococcus sanguis 10556 and Actinomyces oris MG1. To complement the static screen, a flow-based BioFlux microfluidic system screen was also performed under conditions representative of the human oral cavity. Several compounds were found to display biofilm inhibitory activity in all three of the oral bacteria tested. These compounds were also shown to inhibit bioluminescence by Vibrio harveyi and were thus inferred to be quorum sensing (QS) inhibitors. Due to the structural similarity of these compounds to each other, and to key molecules in AI-2 biosynthetic pathways, we propose that these molecules potentially reduce biofilm formation via antagonism of QS or QS-related pathways. This study highlights the potential for a non-antimicrobial-based strategy, focused on AI-2 cell-cell signalling, to control the development of dental plaque. Considering that many bacterial species use AI-2 cell-cell signalling, as well as the increased concern of the use of antimicrobials in healthcare products, such an anti-biofilm approach could also be used to control biofilms in environments beyond the human oral cavity. © 2014 The Society for Applied Microbiology.

  4. The role of counterions in the membrane-disruptive properties of pH-sensitive lysine-based surfactants.

    PubMed

    Nogueira, D R; Mitjans, M; Infante, M R; Vinardell, M P

    2011-07-01

    Surfactants are among the most versatile and widely used excipients in pharmaceuticals. This versatility, together with their pH-responsive membrane-disruptive activity and low toxicity, could also enable their potential application in drug delivery systems. Five anionic lysine-based surfactants which differ in the nature of their counterion were studied. Their capacity to disrupt the cell membrane was examined under a range of pH values, concentrations and incubation times, using a standard hemolysis assay as a model for endosomal membranes. The surfactants showed pH-sensitive hemolytic activity and improved kinetics at the endosomal pH range. Low concentrations resulted in negligible hemolysis at physiological pH and high membrane lytic activity at pH 5.4, which is in the range characteristic of late endosomes. With increasing concentration, the surfactants showed an enhanced capacity to lyse cell membranes, and also caused significant membrane disruption at physiological pH. This observation indicates that, at high concentrations, surfactant behavior is independent of pH. The mechanism of surfactant-mediated membrane destabilization was addressed, and scanning electron microscopy studies were also performed to evaluate the effects of the compounds on erythrocyte morphology as a function of pH. The in vitro cytotoxicity of the surfactants was assessed by MTT and NRU assays with the 3T3 cell line. The influence of different types of counterion on hemolytic activity and the potential applications of these surfactants in drug delivery are discussed. The possibility of using pH-sensitive surfactants for endosome disruption could hold great promise for intracellular drug delivery systems in future therapeutic applications. Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  5. Pyrosequencing analysis yields comprehensive assessment of microbial communities in pilot-scale two-stage membrane biofilm reactors.

    PubMed

    Ontiveros-Valencia, Aura; Tang, Youneng; Zhao, He-Ping; Friese, David; Overstreet, Ryan; Smith, Jennifer; Evans, Patrick; Rittmann, Bruce E; Krajmalnik-Brown, Rosa

    2014-07-01

    We studied the microbial community structure of pilot two-stage membrane biofilm reactors (MBfRs) designed to reduce nitrate (NO3(-)) and perchlorate (ClO4(-)) in contaminated groundwater. The groundwater also contained oxygen (O2) and sulfate (SO4(2-)), which became important electron sinks that affected the NO3(-) and ClO4(-) removal rates. Using pyrosequencing, we elucidated how important phylotypes of each "primary" microbial group, i.e., denitrifying bacteria (DB), perchlorate-reducing bacteria (PRB), and sulfate-reducing bacteria (SRB), responded to changes in electron-acceptor loading. UniFrac, principal coordinate analysis (PCoA), and diversity analyses documented that the microbial community of biofilms sampled when the MBfRs had a high acceptor loading were phylogenetically distant from and less diverse than the microbial community of biofilm samples with lower acceptor loadings. Diminished acceptor loading led to SO4(2-) reduction in the lag MBfR, which allowed Desulfovibrionales (an SRB) and Thiothrichales (sulfur-oxidizers) to thrive through S cycling. As a result of this cooperative relationship, they competed effectively with DB/PRB phylotypes such as Xanthomonadales and Rhodobacterales. Thus, pyrosequencing illustrated that while DB, PRB, and SRB responded predictably to changes in acceptor loading, a decrease in total acceptor loading led to important shifts within the "primary" groups, the onset of other members (e.g., Thiothrichales), and overall greater diversity.

  6. Survival of Deinococcus geothermalis in Biofilms under Desiccation and Simulated Space and Martian Conditions

    NASA Astrophysics Data System (ADS)

    Frösler, Jan; Panitz, Corinna; Wingender, Jost; Flemming, Hans-Curt; Rettberg, Petra

    2017-05-01

    Biofilm formation represents a successful survival strategy for bacteria. In biofilms, cells are embedded in a matrix of extracellular polymeric substances (EPS). As they are often more stress-tolerant than single cells, biofilm cells might survive the conditions present in space and on Mars. To investigate this topic, the bacterium Deinococcus geothermalis was chosen as a model organism due to its tolerance toward desiccation and radiation. Biofilms cultivated on membranes and, for comparison, planktonically grown cells deposited on membranes were air-dried and exposed to individual stressors that included prolonged desiccation, extreme temperatures, vacuum, simulated martian atmosphere, and UV irradiation, and they were exposed to combinations of stressors that simulate space (desiccation + vacuum + UV) or martian (desiccation + Mars atmosphere + UV) conditions. The effect of sulfatic Mars regolith simulant on cell viability during stress was investigated separately. The EPS produced by the biofilm cells contained mainly polysaccharides and proteins. To detect viable but nonculturable (VBNC) cells, cultivation-independent viability indicators (membrane integrity, ATP, 16S rRNA) were determined in addition to colony counts. Desiccation for 2 months resulted in a decrease of culturability with minor changes of membrane integrity in biofilm cells and major loss of membrane integrity in planktonic bacteria. Temperatures between -25°C and +60°C, vacuum, and Mars atmosphere affected neither culturability nor membrane integrity in both phenotypes. Monochromatic (254 nm; ≥1 kJ m-2) and polychromatic (200-400 nm; >5.5 MJ m-2 for planktonic cells and >270 MJ m-2 for biofilms) UV irradiation significantly reduced the culturability of D. geothermalis but did not affect cultivation-independent viability markers, indicating the induction of a VBNC state in UV-irradiated cells. In conclusion, a substantial proportion of the D. geothermalis population remained viable under

  7. Current density reversibly alters metabolic spatial structure of exoelectrogenic anode biofilms

    NASA Astrophysics Data System (ADS)

    Sun, Dan; Cheng, Shaoan; Zhang, Fang; Logan, Bruce E.

    2017-07-01

    Understanding how current densities affect electrogenic biofilm activity is important for wastewater treatment as current densities can substantially decrease at COD concentrations greater than those suitable for discharge to the environment. We examined the biofilm's response, in terms of viability and enzymatic activity, to different current densities using microbial electrolysis cells with a lower (0.7 V) or higher (0.9 V) added voltage to alter current production. Viability was assessed using florescent dyes, with dead cells identified on the basis of dye penetration due to a compromised cell outer-membrane (red), and live cells (intact membrane) fluorescing green. Biofilms operated with 0.7 V produced 2.4 ± 0.2 A m-2, and had an inactive layer near the electrode and a viable layer at the biofilm-solution interface. The lack of cell activity near the electrode surface was confirmed by using an additional dye that fluoresces only with enzymatic activity. Adding 0.9 V increased the current by 61%, and resulted in a single, more homogeneous and active biofilm layer. Switching biofilms between these two voltages produced outcomes associated with the new current rather than the previous biofilm conditions. These findings suggest that maintaining higher current densities will be needed to ensure long-term viability electrogenic biofilms.

  8. Development of a pH sensing membrane electrode based on a new calix[4]arene derivative.

    PubMed

    Kormalı Ertürün, H Elif; Demirel Özel, Ayça; Sayın, Serkan; Yılmaz, Mustafa; Kılıç, Esma

    2015-01-01

    A new pH sensing poly(vinyl chloride) (PVC) membrane electrode was developed by using recently synthesized 5,17-bis(4-benzylpiperidine-1-yl)methyl-25,26,27,28-tetrahydroxy calix[4]arene as an ionophore. The effects of membrane composition, inner filling solution and conditioning solution on the potential response of the proposed pH sensing membrane electrode were investigated. An optimum membrane composition of 3% ionophore, 67% o-nitrophenyl octyl ether (o-NPOE) as plasticizer, 30% PVC was found. The electrode exhibited a near-Nernstian slope of 58.7±1.1 mV pH(-1) in the pH range 1.9-12.7 at 20±1 °C. It showed good selectivity for H(+) ions in the presence of some cations and anions and a longer lifetime of at least 12 months when compared with the other PVC membrane pH electrodes reported in the literature. Having a wide working pH range, it was not only applied as a potentiometric indicator electrode in various acid-base titrations, but also successfully employed in different real samples. It has good reproducibility and repeatability with a response time of 6-7s. Compared to traditional glass pH electrode, it exhibited excellent potentiometric response after being used in fluoride-containing media. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Temporal changes in extracellular polymeric substances on hydrophobic and hydrophilic membrane surfaces in a submerged membrane bioreactor.

    PubMed

    Matar, Gerald; Gonzalez-Gil, Graciela; Maab, Husnul; Nunes, Suzana; Le-Clech, Pierre; Vrouwenvelder, Johannes; Saikaly, Pascal E

    2016-05-15

    Membrane surface hydrophilic modification has always been considered to mitigating biofouling in membrane bioreactors (MBRs). Four hollow-fiber ultrafiltration membranes (pore sizes ∼0.1 μm) differing only in hydrophobic or hydrophilic surface characteristics were operated at a permeate flux of 10 L/m(2h in the same lab-scale MBR fed with synthetic wastewater. In addition, identical membrane modules without permeate production (0 L/m(2h) were operated in the same lab-scale MBR. Membrane modules were autopsied after 1, 10, 20 and 30 days of MBR operation, and total extracellular polymeric substances (EPS) accumulated on the membranes were extracted and characterized in detail using several analytical tools, including conventional colorimetric tests (Lowry and Dubois), liquid chromatography with organic carbon detection (LC-OCD), fluorescence excitation - emission matrices (FEEM), fourier transform infrared (FTIR) and confocal laser scanning microscope (CLSM). The transmembrane pressure (TMP) quickly stabilized with higher values for the hydrophobic membranes than hydrophilic ones. The sulfonated polysulfone (SPSU) membrane had the highest negatively charged membrane surface, accumulated the least amount of foulants and displayed the lowest TMP. The same type of organic foulants developed with time on the four membranes and the composition of biopolymers shifted from protein dominance at early stages of filtration (day 1) towards polysaccharides dominance during later stages of MBR filtration. Nonmetric multidimensional scaling of LC-OCD data showed that biofilm samples clustered according to the sampling event (time) regardless of the membrane surface chemistry (hydrophobic or hydrophilic) or operating mode (with or without permeate flux). These results suggest that EPS composition may not be the dominant parameter for evaluating membrane performance and possibly other parameters such as biofilm thickness, porosity, compactness and structure should be

  10. Outer membrane defect and stronger biofilm formation caused by inactivation of a gene encoding for heptosyltransferase I in Cronobacter sakazakii ATCC BAA-894.

    PubMed

    Wang, L; Hu, X; Tao, G; Wang, X

    2012-05-01

    To investigate the role of lipopolysaccharide (LPS) structure in the stability of outer membrane and the ability of biofilm formation in Cronobacter sakazakii. A C. sakazakii mutant strain LWW02 was constructed by inactivating the gene ESA_04107 encoding for heptosyltransferase I. LPS were purified from LWW02, and changes in their structure were confirmed by thin-layer chromatography and electrospray ionization mass spectrometry. Comparing with the wild-type strain BAA-894, slower growth, higher membrane permeability, higher surface hydrophobicity, stronger ability of autoaggregation and biofilm formation were observed for the mutant strain LWW02. The gene ESA_04107 encodes heptosyltransferase I in C. sakazakii ATCC BAA-894. The cleavage of LPS in C. sakazakii could cause its outer membrane defects and increase its ability to form biofilms. The study is important for understanding the pathogenic mechanism and efficient control of C. sakazakii. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  11. H2 conversion in the presence of O2 as performed by the membrane-bound [NiFe]-hydrogenase of Ralstonia eutropha.

    PubMed

    Lenz, Oliver; Ludwig, Marcus; Schubert, Torsten; Bürstel, Ingmar; Ganskow, Stefanie; Goris, Tobias; Schwarze, Alexander; Friedrich, Bärbel

    2010-04-26

    [NiFe]-hydrogenases catalyze the oxidation of H(2) to protons and electrons. This reversible reaction is based on a complex interplay of metal cofactors including the Ni-Fe active site and several [Fe-S] clusters. H(2) catalysis of most [NiFe]-hydrogenases is sensitive to dioxygen. However, some bacteria contain hydrogenases that activate H(2) even in the presence of O(2). There is now compelling evidence that O(2) affects hydrogenase on three levels: 1) H(2) catalysis, 2) hydrogenase maturation, and 3) H(2)-mediated signal transduction. Herein, we summarize the genetic, biochemical, electrochemical, and spectroscopic properties related to the O(2) tolerance of hydrogenases resident in the facultative chemolithoautotroph Ralstonia eutropha H16. A focus is given to the membrane-bound [NiFe]-hydogenase, which currently represents the best-characterized member of O(2)-tolerant hydrogenases.

  12. Dissolved molecular hydrogen (H2) in Peritoneal Dialysis (PD) solutions preserves mesothelial cells and peritoneal membrane integrity.

    PubMed

    Nakayama, Masaaki; Zhu, Wan-Jun; Watanabe, Kimio; Gibo, Ayano; Sherif, Ali M; Kabayama, Shigeru; Ito, Sadayoshi

    2017-10-31

    Peritoneal dialysis (PD) is used as renal replacement therapy in patients with end-stage kidney disease. However, peritoneal membrane failure remains problematic and constitutes a critical cause of PD discontinuation. Recent studies have revealed the unique biological action of molecular hydrogen (H 2 ) as an anti-oxidant, which ameliorates tissue injury. In the present study, we aimed to examine the effects of H 2 on the peritoneal membrane of experimental PD rats. Eight-week-old male Sprague-Dawley rats were divided into the following groups (n = 8-11 each) receiving different test solutions: control group (no treatment), PD group (commercially available lactate-based neutral 2.5% glucose PD solution), and H 2 PD group (PD solution with dissolved H 2 at 400 ppb). Furthermore, the influence of iron (FeCl 3 : 5 μM: inducer of oxidative cellular injury) in the respective PD solutions was also examined (Fe-PD and Fe-H 2 PD groups). The H 2 PD solution was manufactured by bathing a PD bag in H 2 -oversaturated water created by electrolysis of the water. Twenty mL of the test solutions were intraperitoneally injected once a day for 10 days. Parietal peritoneum samples and cells collected from the peritoneal surface following treatment with trypsin were subjected to analysis. In the PD group as compared to controls, a mild but significant sub-mesothelial thickening was observed, with increase in the number of cells in the peritoneal surface tissue that were positive for apoptosis, proliferation and vimentin, as seen by immunostaining. There were significantly fewer of such changes in the H 2 PD group, in which there was a dominant presence of M2 (CD163+) macrophages in the peritoneum. The Fe-PD group showed a significant loss of mesothelial cells with sub-mesothelial thickening, these changes being ameliorated in the Fe-H 2 PD group. H 2 -dissolved PD solutions could preserve mesothelial cells and peritoneal membrane integrity in PD rats. Clinical application of H

  13. Discovering Biofilms: Inquiry-Based Activities for the Classroom

    ERIC Educational Resources Information Center

    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…

  14. Sequence of colonization determines the composition of mixed biofilms by Escherichia coli O157:H7 and O111:H8 strains

    USDA-ARS?s Scientific Manuscript database

    Bacterial biofilms are one of the potential sources of cross-contamination in food processing environments. Shiga-toxin producing Escherichia coli (STEC) O157:H7 and O111:H8 are important foodborne pathogens capable of forming biofilms, and the coexistence of these two STEC serotypes has been detec...

  15. Nanocomposite membranes based on polybenzimidazole and ZrO2 for high-temperature proton exchange membrane fuel cells.

    PubMed

    Nawn, Graeme; Pace, Giuseppe; Lavina, Sandra; Vezzù, Keti; Negro, Enrico; Bertasi, Federico; Polizzi, Stefano; Di Noto, Vito

    2015-04-24

    Owing to the numerous benefits obtained when operating proton exchange membrane fuel cells at elevated temperature (>100 °C), the development of thermally stable proton exchange membranes that demonstrate conductivity under anhydrous conditions remains a significant goal for fuel cell technology. This paper presents composite membranes consisting of poly[2,2'-(m-phenylene)-5,5'-bibenzimidazole] (PBI4N) impregnated with a ZrO2 nanofiller of varying content (ranging from 0 to 22 wt %). The structure-property relationships of the acid-doped and undoped composite membranes have been studied using thermogravimetric analysis, differential scanning calorimetry, dynamic mechanical analysis, wide-angle X-ray scattering, infrared spectroscopy, and broadband electrical spectroscopy. Results indicate that the level of nanofiller has a significant effect on the membrane properties. From 0 to 8 wt %, the acid uptake as well as the thermal and mechanical properties of the membrane increase. As the nanofiller level is increased from 8 to 22 wt % the opposite effect is observed. At 185 °C, the ionic conductivity of [PBI4N(ZrO2 )0.231 ](H3 PO4 )13 is found to be 1.04×10(-1)  S cm(-1) . This renders membranes of this type promising candidates for use in high-temperature proton exchange membrane fuel cells. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Biofilm three-dimensional architecture influences in situ pH distribution pattern on the human enamel surface

    PubMed Central

    Xiao, Jin; Hara, Anderson T; Kim, Dongyeop; Zero, Domenick T; Koo, Hyun; Hwang, Geelsu

    2017-01-01

    To investigate how the biofilm three-dimensional (3D) architecture influences in situ pH distribution patterns on the enamel surface. Biofilms were formed on human tooth enamel in the presence of 1% sucrose or 0.5% glucose plus 0.5% fructose. At specific time points, biofilms were exposed to a neutral pH buffer to mimic the buffering of saliva and subsequently pulsed with 1% glucose to induce re-acidification. Simultaneous 3D pH mapping and architecture of intact biofilms was performed using two-photon confocal microscopy. The enamel surface and mineral content characteristics were examined successively via optical profilometry and microradiography analyses. Sucrose-mediated biofilm formation created spatial heterogeneities manifested by complex networks of bacterial clusters (microcolonies). Acidic regions (pH<5.5) were found only in the interior of microcolonies, which impedes rapid neutralization (taking more than 120 min for neutralization). Glucose exposure rapidly re-created the acidic niches, indicating formation of diffusion barriers associated with microcolonies structure. Enamel demineralization (white spots), rougher surface, deeper lesion and more mineral loss appeared to be associated with the localization of these bacterial clusters at the biofilm-enamel interface. Similar 3D architecture was observed in plaque-biofilms formed in vivo in the presence of sucrose. The formation of complex 3D architectures creates spatially heterogeneous acidic microenvironments in close proximity of enamel surface, which might correlate with the localized pattern of the onset of carious lesions (white spot like) on teeth. PMID:28452377

  17. H-NOX Regulation of c-di-GMP Metabolism and Biofilm Formation in Legionella pneumophila

    PubMed Central

    Carlson, Hans K.; Vance, Russell E.; Marletta, Michael A.

    2010-01-01

    Summary Heme Nitric oxide/OXygen (H-NOX) domains are a family of hemoprotein sensors that are widespread in bacterial genomes, but limited information is available on their function. Legionella pneumophila is the only prokaryote found thus far to encode two H-NOX proteins. This paper presents data supporting a role for one of the Legionella pneumophila H-NOXs in the regulation of biofilm formation. In summary: (i) unmarked deletions in the hnox1 gene do not affect growth rate in liquid culture or replication in permissive macrophages; (ii) theΔhnox1 strain displays a hyper-biofilm phenotype; (iii) the gene adjacent to hnox1 is a GGDEF-EAL protein, lpg1057, and overexpression in Legionella pneumophila of this protein, or the well-studied diguanylate cyclase, vca0956, results in a hyper-biofilm phenotype; (iv) the Lpg1057 protein displays diguanylate cyclase activity in vitro and this activity is inhibited by the Hnox1 protein in the Fe(II)-NO ligation state, but not the Fe(II) unligated state; (v) consistent with the Hnox1 regulation of Lpg1057, unmarked deletions of lpg1057 in theΔhnox1 background results in reversion of the hyper-biofilm phenotype back to wild-type biofilm levels. Taken together, these results suggest a role for hnox1 in regulating c-di-GMP production by lpg1057 and biofilm formation in response to NO. PMID:20572940

  18. Gypsum (CaSO4·2H2O) Scaling on Polybenzimidazole and Cellulose Acetate Hollow Fiber Membranes under Forward Osmosis

    PubMed Central

    Chen, Si Cong; Su, Jincai; Fu, Feng-Jiang; Mi, Baoxia; Chung, Tai-Shung

    2013-01-01

    We have examined the gypsum (CaSO4·2H2O) scaling phenomena on membranes with different physicochemical properties in forward osmosis (FO) processes. Three hollow fiber membranes made of (1) cellulose acetate (CA), (2) polybenzimidazole (PBI)/polyethersulfone (PES) and (3) PBI-polyhedral oligomeric silsesquioxane (POSS)/polyacrylonitrile (PAN) were studied. For the first time in FO processes, we have found that surface ionic interactions dominate gypsum scaling on the membrane surface. A 70% flux reduction was observed on negatively charged CA and PBI membrane surfaces, due to strong attractive forces. The PBI membrane surface also showed a slightly positive charge at a low pH value of 3 and exhibited a 30% flux reduction. The atomic force microscopy (AFM) force measurements confirmed a strong repulsive force between gypsum and PBI at a pH value of 3. The newly developed PBI-POSS/PAN membrane had ridge morphology and a contact angle of 51.42° ± 14.85° after the addition of hydrophilic POSS nanoparticles and 3 min thermal treatment at 95 °C. Minimal scaling and an only 1.3% flux reduction were observed at a pH value of 3. Such a ridge structure may reduce scaling by not providing a locally flat surface to the crystallite at a pH value of 3; thus, gypsum would be easily washed away from the surface. PMID:24957062

  19. Physiological differentiation within a single-species biofilm fueled by serpentinization.

    PubMed

    Brazelton, William J; Mehta, Mausmi P; Kelley, Deborah S; Baross, John A

    2011-01-01

    Carbonate chimneys at the Lost City hydrothermal field are coated in biofilms dominated by a single phylotype of archaea known as Lost City Methanosarcinales. In this study, we have detected surprising physiological complexity in single-species biofilms, which is typically indicative of multispecies biofilm communities. Multiple cell morphologies were visible within the biofilms by transmission electron microscopy, and some cells contained intracellular membranes that may facilitate methane oxidation. Both methane production and oxidation were detected at 70 to 80°C and pH 9 to 10 in samples containing the single-species biofilms. Both processes were stimulated by the presence of hydrogen (H(2)), indicating that methane production and oxidation are part of a syntrophic interaction. Metagenomic data included a sequence encoding AMP-forming acetyl coenzyme A synthetase, indicating that acetate may play a role in the methane-cycling syntrophy. A wide range of nitrogen fixation genes were also identified, many of which were likely acquired via lateral gene transfer (LGT). Our results indicate that cells within these single-species biofilms may have differentiated into multiple physiological roles to form multicellular communities linked by metabolic interactions and LGT. Communities similar to these Lost City biofilms are likely to have existed early in the evolution of life, and we discuss how the multicellular characteristics of ancient hydrogen-fueled biofilm communities could have stimulated ecological diversification, as well as unity of biochemistry, during the earliest stages of cellular evolution. Our previous work at the Lost City hydrothermal field has shown that its carbonate chimneys host microbial biofilms dominated by a single uncultivated "species" of archaea. In this paper, we integrate evidence from these previous studies with new data on the metabolic activity and cellular morphology of these archaeal biofilms. We conclude that the archaeal biofilm

  20. Transcriptomic analysis for genetic mechanisms of the factors related to biofilm formation in Escherichia coli O157:H7.

    PubMed

    Lee, Jin-Hyung; Kim, Yong-Guy; Cho, Moo Hwan; Wood, Thomas K; Lee, Jintae

    2011-04-01

    Two lineages of enterohemorrhagic Escherichia coli O157:H7 (EDL933, Stx1(+) and Stx2(+)) and 86-24 (Stx2(+)) were investigated to determine the genetic basis of biofilm formation on abiotic surfaces. Strain EDL933 formed a robust biofilm while strain 86-24 formed almost no biofilm on either polystyrene plates or polyethylene tubes. Whole-transcriptome profiles of EDL933 versus 86-24 revealed that in the strong biofilm-forming strain, genes involved in curli biosynthesis and cellulose production were significantly induced, whereas genes involved in indole signaling were most repressed. Additionally, 49 phage genes were highly induced and repressed between the two strains. Curli assays using Congo red plates and scanning electron microscopy corroborated the microarray data as the EDL933 strain produced a large amount of curli, while strain 86-24 formed much less curli. Moreover, EDL933 produced 19-fold more cellulose than 86-24, and indole production in EDL933 was two times lower than that of the strain 86-24. Therefore, it appears E. coli O157:H7 EDL933 produces more biofilm because of its increased curli and cellulose production and reduced indole production.

  1. Purification of plant plasma membranes by two-phase partitioning and measurement of H+ pumping.

    PubMed

    Lund, Anette; Fuglsang, Anja Thoe

    2012-01-01

    Purification of plasma membranes by two-phase partitioning is based on the separation of microsomal membranes, dependent on their surface hydrophobicity. Here we explain the purification of plasma membranes from a relatively small amount of material (7-30 g). The fluorescent probe ACMA (9-amino-6-chloro-2-metoxyacridine) accumulates inside the vesicles upon protonation. Quenching of ACMA in the solution corresponds to the H(+) transport across the plasma membrane. Before running the assay, the plasma membranes are incubated with the detergent Brij-58 in order to create inside-out vesicles.Purification of plasma membranes by two-phase partitioning is based on the separation of microsomal membranes, dependent on their surface hydrophobicity. Here we explain the purification of plasma membranes from a relatively small amount of material (7-30 g). The fluorescent probe ACMA (9-amino-6-chloro-2-metoxyacridine) accumulates inside the vesicles upon protonation. Quenching of ACMA in the solution corresponds to the H(+) transport across the plasma membrane. Before running the assay, the plasma membranes are incubated with the detergent Brij-58 in order to create inside-out vesicles.

  2. Biofilm formation by virulent and non-virulent strains of Haemophilus parasuis.

    PubMed

    Bello-Ortí, Bernardo; Deslandes, Vincent; Tremblay, Yannick D N; Labrie, Josée; Howell, Kate J; Tucker, Alexander W; Maskell, Duncan J; Aragon, Virginia; Jacques, Mario

    2014-11-27

    Haemophilus parasuis is a commensal bacterium of the upper respiratory tract of healthy pigs. It is also the etiological agent of Glässer's disease, a systemic disease characterized by polyarthritis, fibrinous polyserositis and meningitis, which causes high morbidity and mortality in piglets. The aim of this study was to evaluate biofilm formation by well-characterized virulent and non-virulent strains of H. parasuis. We observed that non-virulent strains isolated from the nasal cavities of healthy pigs formed significantly (p < 0.05) more biofilms than virulent strains isolated from lesions of pigs with Glässer's disease. These differences were observed when biofilms were formed in microtiter plates under static conditions or formed in the presence of shear force in a drip-flow apparatus or a microfluidic system. Confocal laser scanning microscopy using different fluorescent probes on a representative subset of strains indicated that the biofilm matrix contains poly-N-acetylglucosamine, proteins and eDNA. The biofilm matrix was highly sensitive to degradation by proteinase K. Comparison of transcriptional profiles of biofilm and planktonic cells of the non-virulent H. parasuis F9 strain revealed a significant number of up-regulated membrane-related genes in biofilms, and genes previously identified in Actinobacillus pleuropneumoniae biofilms. Our data indicate that non-virulent strains of H. parasuis have the ability to form robust biofilms in contrast to virulent, systemic strains. Biofilm formation might therefore allow the non-virulent strains to colonize and persist in the upper respiratory tract of pigs. Conversely, the planktonic state of the virulent strains might allow them to disseminate within the host.

  3. Physiological Differentiation within a Single-Species Biofilm Fueled by Serpentinization

    PubMed Central

    Brazelton, William J.; Mehta, Mausmi P.; Kelley, Deborah S.; Baross, John A.

    2011-01-01

    ABSTRACT Carbonate chimneys at the Lost City hydrothermal field are coated in biofilms dominated by a single phylotype of archaea known as Lost City Methanosarcinales. In this study, we have detected surprising physiological complexity in single-species biofilms, which is typically indicative of multispecies biofilm communities. Multiple cell morphologies were visible within the biofilms by transmission electron microscopy, and some cells contained intracellular membranes that may facilitate methane oxidation. Both methane production and oxidation were detected at 70 to 80°C and pH 9 to 10 in samples containing the single-species biofilms. Both processes were stimulated by the presence of hydrogen (H2), indicating that methane production and oxidation are part of a syntrophic interaction. Metagenomic data included a sequence encoding AMP-forming acetyl coenzyme A synthetase, indicating that acetate may play a role in the methane-cycling syntrophy. A wide range of nitrogen fixation genes were also identified, many of which were likely acquired via lateral gene transfer (LGT). Our results indicate that cells within these single-species biofilms may have differentiated into multiple physiological roles to form multicellular communities linked by metabolic interactions and LGT. Communities similar to these Lost City biofilms are likely to have existed early in the evolution of life, and we discuss how the multicellular characteristics of ancient hydrogen-fueled biofilm communities could have stimulated ecological diversification, as well as unity of biochemistry, during the earliest stages of cellular evolution. PMID:21791580

  4. Curli fimbriae are conditionally required in Escherichia coli O157:H7 for initial attachment and biofilm formation.

    PubMed

    Carter, Michelle Qiu; Louie, Jacqueline W; Feng, Doris; Zhong, Wayne; Brandl, Maria T

    2016-08-01

    Several species of enteric pathogens produce curli fimbriae, which may affect their interaction with surfaces and other microbes in nonhost environments. Here we used two Escherichia coli O157:H7 outbreak strains with distinct genotypes to understand the role of curli in surface attachment and biofilm formation in several systems relevant to fresh produce production and processing. Curli significantly enhanced the initial attachment of E. coli O157:H7 to spinach leaves and stainless steel surfaces by 5-fold. Curli was also required for E. coli O157:H7 biofilm formation on stainless steel and enhanced biofilm production on glass by 19-27 fold in LB no-salt broth. However, this contribution was not observed when cells were grown in sterile spinach lysates. Furthermore, both strains of E. coli O157:H7 produced minimal biofilms on polypropylene in LB no-salt broth but considerable amounts in spinach lysates. Under the latter conditions, curli appeared to slightly increase biofilm production. Importantly, curli played an essential role in the formation of mixed biofilm by E. coli O157:H7 and plant-associated microorganisms in spinach leaf washes, as revealed by confocal microscopy. Little or no E. coli O157:H7 biofilms were detected at 4 °C, supporting the importance of temperature control in postharvest and produce processing environments. Published by Elsevier Ltd.

  5. Micafungin triggers caspase-dependent apoptosis in Candida albicans and Candida parapsilosis biofilms, including caspofungin non-susceptible isolates

    PubMed Central

    Shirazi, F; Kontoyiannis, DP

    2015-01-01

    Candida biofilms play an important role in infections associated with medical devices and are resistant to antifungals. We hypothesized that the echinocandin micafungin (MICA) exerts an enhanced antifungal activity against caspofungin (CAS)-susceptible (CAS-S) and CAS–non-susceptible (CAS-NS) Candida albicans and Candida parapsilosis which is at least in part through apoptosis, even in the biofilm environment. Apoptosis was characterized by detecting reactive oxygen species (ROS) accumulation, depolarization of mitochondrial membrane potential (MMP), DNA fragmentation, lack of plasma membrane integrity, and metacaspase activation following exposure of Candida biofilm to MICA for 3h at 37°C in RPMI 1640 medium. The minimum inhibitory concentration was higher for CAS (2.0–16.0 μg/mL) than for MICA (1.0–8.0 μg/mL) for Candida biofilms. Elevated intracellular ROS levels and depolarization of MMP was evident in CAS-S C. albicans (3.0–4.2 fold) and C. parapsilosis (4.8–5.4 fold) biofilms compared with CAS-NS (1.2 fold) after exposure to MICA (0.25x-1xMIC). Elevated intracellular ROS levels and depolarization of MMP was evident in CAS-S C. albicans (3.0–4.2 fold) and C. parapsilosis (4.8–5.4 fold) biofilms compared with CAS-NS (1.2 fold) after exposure to MICA (0.25x-1xMIC). Finally higher ß-1, 3 glucan levels were seen in sessile cells compared to planktonic cells, especially in CAS-NS strains. MICA treatment might induce a metacaspase-dependent apoptotic process in biofilms of both CAS-S C. albicans and C. parapsilosis, and to some degree in CAS-NS strains. PMID:26065323

  6. Method for forming H2-permselective oxide membranes

    DOEpatents

    Gavalas, George R.; Nam, Suk Woo; Tsapatsis, Michael; Kim, Soojin

    1995-01-01

    Methods of forming permselective oxide membranes that are highly selective to permeation of hydrogen by chemical deposition of reactants in the pore of porous tubes, such as Vycor.TM. glass or Al.sub.2 O.sub.3 tubes. The porous tubes have pores extending through the tube wall. The process involves forming a stream containing a first reactant of the formula RX.sub.n, wherein R is silicon, titanium, boron or aluminum, X is chlorine, bromine or iodine, and n is a number which is equal to the valence of R; and forming another stream containing water vapor as the second reactant. Both of the reactant streams are passed along either the outside or the inside surface of a porous tube and the streams react in the pores of the porous tube to form a nonporous layer of R-oxide in the pores. The membranes are formed by the hydrolysis of the respective halides. In another embodiment, the first reactant stream contains a first reactant having the formula SiH.sub.n Cl.sub.4-n where n is 1, 2 or 3; and the second reactant stream contains water vapor and oxygen. In still another embodiment the first reactant stream containing a first reactant selected from the group consisting of Cl.sub.3 SiOSiCl.sub.3, Cl.sub.3 SiOSiCl.sub.2 OSiCl.sub.3, and mixtures thereof and the second reactant stream contains water vapor. In still another embodiment, membrane formation is carried out by an alternating flow deposition method. This involves a sequence of cycles, each cycle comprising introduction of the halide-containing stream and allowance of a specific time for reaction followed by purge and flow of the water vapor containing stream for a specific length of time. In all embodiments the nonporous layers formed are selectively permeable to hydrogen.

  7. Heterologous Expression of sahH Reveals That Biofilm Formation Is Autoinducer-2-independent in Streptococcus sanguinis but Is Associated with an Intact Activated Methionine Cycle*

    PubMed Central

    Redanz, Sylvio; Standar, Kerstin; Podbielski, Andreas; Kreikemeyer, Bernd

    2012-01-01

    Numerous studies have claimed deleterious effects of LuxS mutation on many bacterial phenotypes, including bacterial biofilm formation. Genetic complementation mostly restored the observed mutant phenotypes to WT levels, leading to the postulation that quorum sensing via a family of molecules generically termed autoinducer-2 (AI-2) is essential for many phenotypes. Because LuxS mutation has dual effects, this hypothesis needs to be investigated into the details for each bacterial species. In this study we used S. sanguinis SK36 as a model biofilm bacterium and employed physiological characterization and transcriptome approaches on WT and luxS-deficient strains, in combination with chemical, luxS, and sahH complementation experiments. SahH enables a direct conversion of SAH to homocysteine and thereby restores the activated methionine cycle in a luxS-negative background without formation of the AI-2 precursor 4,5-dihydroxy-2,3-pentanedione. With this strategy we were able to dissect the individual contribution of LuxS and AI-2 activity in detail. Our data revealed that S. sanguinis biofilm formation is independent from AI-2 substance pools and is rather supported by an intact activated methyl cycle. Of 216 differentially transcribed genes in the luxS mutant, 209 were restored by complementation with a gene encoding the S-adenosylhomocysteine hydrolase. Only nine genes, mainly involved in natural competence, were directly affected by the AI-2 quorum-sensing substance pool. Cumulatively, this suggested that biofilm formation in S. sanguinis is not under control of AI-2. Our study suggests that previously evaluated LuxS mutants in other species need to be revisited to resolve the precise contribution of AI-2 substance pools and the methionine pathways. PMID:22942290

  8. Microbial community in biofilm on membrane surface of submerged MBR: effect of in-line cleaning chemical agent.

    PubMed

    Lim, B R; Ahn, K H; Song, K G; Cho, J W

    2005-01-01

    The objective of this study was to investigate the change in microbial community pattern with the effect of cleaning agent using a quinone profile that is used for membrane in-line chemical cleaning in SMBR. The dominant quinone types of biofilm were ubiquinone (UQs)-8, -10, followed by menaquinone (MKs)-8(H4), -7 and UQ-9, but those of suspended microorganisms were UQ-8, UQ-10 followed by MKs-8(H4), -7 and -11. Both UQ and MK contents decreased with increasing NaCIO dosage and it seems that there is more resistance from UQ compared to MK. In addition, COD and DOC concentrations increased with increasing NaClO dosage up to 0.05 g-NaCIO/g-SS. The organic degradation performance of the microbial community in the presence of NaClO was impaired. The present study suggested that larger added amounts of NaClO caused an inhibition of organic degradation and cell lysis.

  9. Inhibition on Candida albicans biofilm formation using divalent cation chelators (EDTA).

    PubMed

    Ramage, Gordon; Wickes, Brian L; López-Ribot, José L

    2007-12-01

    Candida albicans can readily form biofilms on both inanimate and biological surfaces. In this study we investigated a means of inhibiting biofilm formation using EDTA (Ethylenediaminetetra-acetic acid), a divalent cation chelating agent, which has been shown to affect C. albicans filamentation. Candida albicans biofilms were formed in 96-well microtitre plates. Cells were allowed to adhere for 1, 2, and 4 h at 37 degrees C, washed in PBS, and then treated with different concentrations of EDTA (0, 2.5, 25, and 250 mM). EDTA was also added to the standardized suspension prior to adding to the microtiter plate and to a preformed 24 h biofilm. All plates were then incubated at 37 degrees C for an additional 24 h to allow for biofilm formation. The extent and characteristics of biofilm formation were then microscopically assessed and with a semi-quantitative colorimetric technique based on the use of an XTT-reduction assay. Northern blot analysis of the hyphal wall protein (HWP1) expression was also monitored in planktonic and biofilm cells treated with EDTA. Microscopic analysis and colorimetric readings revealed that filamentation and biofilm formation were inhibited by EDTA in a concentration dependent manner. However, preformed biofilms were minimally affected by EDTA (maximum of 31% reduction at 250 mM). The HWP1 gene expression was reduced in EDTA-treated planktonic and biofilm samples. These results indicate that EDTA inhibits C. albicans biofilm formation are most likely through its inhibitory effect on filamentation and indicates the potential therapeutic effects of EDTA. This compound may serve a non-toxic means of preventing biofilm formation on infections with a C. albicans biofilm etiology.

  10. Biofilm formation enhances Helicobacter pylori survivability in vegetables.

    PubMed

    Ng, Chow Goon; Loke, Mun Fai; Goh, Khean Lee; Vadivelu, Jamuna; Ho, Bow

    2017-04-01

    To date, the exact route and mode of transmission of Helicobacter pylori remains elusive. The detection of H. pylori in food using molecular approaches has led us to postulate that the gastric pathogen may survive in the extragastric environment for an extended period. In this study, we show that H. pylori prolongs its survival by forming biofilm and micro-colonies on vegetables. The biofilm forming capability of H. pylori is both strain and vegetable dependent. H. pylori strains were classified into high and low biofilm formers based on their highest relative biofilm units (BU). High biofilm formers survived longer on vegetables compared to low biofilm formers. The bacteria survived better on cabbage compared to other vegetables tested. In addition, images captured on scanning electron and confocal laser scanning microscopes revealed that the bacteria were able to form biofilm and reside as micro-colonies on vegetable surfaces, strengthening the notion of possible survival of H. pylori on vegetables for an extended period of time. Taken together, the ability of H. pylori to form biofilm on vegetables (a common food source for human) potentially plays an important role in its survival, serving as a mode of transmission of H. pylori in the extragastric environment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. (Super)hydrophobic coating of orthodontic dental devices and reduction of early oral biofilm retention.

    PubMed

    Oliveira, Adauê S; Kaizer, Marina R; Azevedo, Marina S; Ogliari, Fabrício A; Cenci, Maximiliano S; Moraes, Rafael R

    2015-11-03

    This study was designed to apply (super)hydrophobic crosslinked coatings by means of a sol-gel process on the surface of orthodontic devices and investigate the potential effect of these coatings in reducing the early retention of oral biofilm. Two organosilane-based hydrophobic solutions (HSs) were prepared containing hexadecyltrimethoxysilane diluted in ethanol (HS1) or 1H, 1H, 2H, 2H-perfluorodecyltriethoxysilane diluted in dimethyl sulfoxide (HS2). Stainless steel plates and ceramic discs were coated with HS1 or HS2 and heated at 150 °C for 2 h for condensation of a crosslinked SiO x network. Organosilane coatings were applied after previous, or no, surface sandblasting. Commercial stainless steel and ceramic brackets were used to evaluate oral biofilm retention after 12 h or 24 h of biofilm growth, using a microcosm model with human saliva as the inoculum. Surface roughness analysis (Ra, μm) indicated that sandblasting associated with organosilane coatings increased roughness for stainless steel brackets only. Analysis of the water contact angle showed that the stainless steel surface treated with HS1 was hydrophobic (~123°), while the ceramic surface treated with HS2 was superhydrophobic (~155°). Biofilm retention after 24 h was significantly lower in groups treated with hydrophobic coatings. An exponential reduction in biofilm accumulation was associated with increased water contact angle for both stainless steel and ceramic at 24 h. Application of (super)hydrophobic coatings on the surface of stainless steel and ceramic orthodontic devices might reduce the retention of oral biofilm.

  12. Sublethal concentrations of silver nanoparticles affect the mechanical stability of biofilms.

    PubMed

    Grün, Alexandra Y; Meier, Jutta; Metreveli, George; Schaumann, Gabriele E; Manz, Werner

    2016-12-01

    Bacterial biofilms are most likely confronted with silver nanoparticles (Ag NPs) as a pollutant stressor in aquatic systems. In this study, biofilms of Aquabacterium citratiphilum were exposed for 20 h to 30 and 70 nm citrate stabilized Ag NPs in low-dose concentrations ranging from 600 to 2400 μg l -1 , and the Ag NP-mediated effects on descriptive, structural, and functional biofilm characteristics, including viability, protein content, architecture, and mechanical stability, were investigated. Viability, based on the bacterial cell membrane integrity of A. citratiphilum, as determined by epifluorescence microscopy, remained unaffected after Ag NP exposure. Moreover, in contrast to information in the current literature, protein contents of cells and extracellular polymeric substances (EPS) and biofilm architecture, including dry mass, thickness, and density, were not significantly impacted by exposure to Ag NPs. However, the biofilms themselves served as effective sinks for Ag NPs, exhibiting enrichment factors from 5 to 8. Biofilms showed a greater capacity to accumulate 30 nm sized Ag NPs than 70 nm Ag NPs. Furthermore, Ag NPs significantly threatened the mechanical stability of biofilms, as determined by a newly developed assay. For 30 nm Ag NPs, the mechanical stability of biofilms decreased as the Ag NP concentrations applied to them increased. In contrast, 70 nm Ag NPs produced a similar decrease in mechanical stability for each applied concentration. Overall, this finding demonstrates that exposure to Ag NPs triggers remarkable changes in biofilm adhesion and/or cohesiveness. Because of biofilm-mediated ecological services, this response raises environmental concerns regarding Ag NP release into freshwater systems, even in sublethal concentrations.

  13. Membranes, methods of making membranes, and methods of separating gases using membranes

    DOEpatents

    Ho, W. S. Winston

    2012-10-02

    Membranes, methods of making membranes, and methods of separating gases using membranes are provided. The membranes can include at least one hydrophilic polymer, at least one cross-linking agent, at least one base, and at least one amino compound. The methods of separating gases using membranes can include contacting a gas stream containing at least one of CO.sub.2, H.sub.2S, and HCl with one side of a nonporous and at least one of CO.sub.2, H.sub.2S, and HCl selectively permeable membrane such that at least one of CO.sub.2, H.sub.2S, and HCl is selectively transported through the membrane.

  14. Membrane-destabilizing activity of pH-responsive cationic lysine-based surfactants: role of charge position and alkyl chain length.

    PubMed

    Nogueira, Daniele Rubert; Mitjans, Montserrat; Morán, M Carmen; Pérez, Lourdes; Vinardell, M Pilar

    2012-09-01

    Many strategies for treating diseases require the delivery of drugs into the cell cytoplasm following internalization within endosomal vesicles. Thus, compounds triggered by low pH to disrupt membranes and release endosomal contents into the cytosol are of particular interest. Here, we report novel cationic lysine-based surfactants (hydrochloride salts of N(ε)- and N(α)-acyl lysine methyl ester) that differ in the position of the positive charge and the length of the alkyl chain. Amino acid-based surfactants could be promising novel biomaterials in drug delivery systems, given their biocompatible properties and low cytotoxic potential. We examined their ability to disrupt the cell membrane in a range of pH values, concentrations and incubation times, using a standard hemolysis assay as a model of endosomal membranes. Furthermore, we addressed the mechanism of surfactant-mediated membrane destabilization, including the effects of each surfactant on erythrocyte morphology as a function of pH. We found that only surfactants with the positive charge on the α-amino group of lysine showed pH-sensitive hemolytic activity and improved kinetics within the endosomal pH range, indicating that the positive charge position is critical for pH-responsive behavior. Moreover, our results showed that an increase in the alkyl chain length from 14 to 16 carbon atoms was associated with a lower ability to disrupt cell membranes. Knowledge on modulating surfactant-lipid bilayer interactions may help us to develop more efficient biocompatible amino acid-based drug delivery devices.

  15. Evaluation of micropollutant removal and fouling reduction in a hybrid moving bed biofilm reactor-membrane bioreactor system.

    PubMed

    Luo, Yunlong; Jiang, Qi; Ngo, Huu H; Nghiem, Long D; Hai, Faisal I; Price, William E; Wang, Jie; Guo, Wenshan

    2015-09-01

    A hybrid moving bed biofilm reactor-membrane bioreactor (MBBR-MBR) system and a conventional membrane bioreactor (CMBR) were compared in terms of micropollutant removal efficiency and membrane fouling propensity. The results show that the hybrid MBBR-MBR system could effectively remove most of the selected micropollutants. By contrast, the CMBR system showed lower removals of ketoprofen, carbamazepine, primidone, bisphenol A and estriol by 16.2%, 30.1%, 31.9%, 34.5%, and 39.9%, respectively. Mass balance calculations suggest that biological degradation was the primary removal mechanism in the MBBR-MBR system. During operation, the MBBR-MBR system exhibited significantly slower fouling development as compared to the CMBR system, which could be ascribed to the wide disparity in the soluble microbial products (SMP) levels between MBBR-MBR (4.02-6.32 mg/L) and CMBR (21.78 and 33.04 mg/L). It is evident that adding an MBBR process prior to MBR treatment can not only enhance micropollutant elimination but also mitigate membrane fouling. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Soluble factors from biofilm of Candida albicans and Staphylococcus aureus promote cell death and inflammatory response.

    PubMed

    de Carvalho Dias, Kassia; Barbugli, Paula Aboud; de Patto, Fernanda; Lordello, Virginia Barreto; de Aquino Penteado, Letícia; Medeiros, Alexandra Ivo; Vergani, Carlos Eduardo

    2017-06-30

    The objective of this study was to better understand the effects of soluble factors from biofilm of single- and mixed-species Candida albicans (C. albicans) and methicillin-sensitive Staphylococcus aureus (MSSA) cultures after 36 h in culture on keratinocytes (NOK-si and HaCaT) and macrophages (J774A.1). Soluble factors from biofilms of C. albicans and MSSA were collected and incubated with keratinocytes and macrophages, which were subsequently evaluated by cell viability assays (MTT). Lactate dehydrogenase (LDH) enzyme release was measured to assess cell membrane damage to keratinocytes. Cells were analysed by brightfield microscopy after 2 and 24 h of exposure to the soluble factors from biofilm. Cell death was detected by labelling apoptotic cells with annexin V and necrotic cells with propidium iodide (PI) and was visualized via fluorescence microscopy. Soluble factors from biofilm were incubated with J774A.1 cells for 24 h; the subsequent production of NO and the cytokines IL-6 and TNF-α was measured by ELISA. The cell viability assays showed that the soluble factors of single-species C. albicans cultures were as toxic as the soluble factors from biofilm of mixed cultures, whereas the soluble factors of MSSA cultures were less toxic than those of C. albicans or mixed cultures. The soluble factors from biofilm of mixed cultures were the most toxic to the NOK-si and HaCaT cells, as confirmed by analyses of PI labelling and cell morphology. Soluble factors from biofilm of single-species MSSA and mixed-species cultures induced the production of IL-6, NO and TNF-α by J744A.1 macrophages. The production of IL-6 and NO induced by the soluble factors from biofilm of mixed cultures was lower than that induced by the soluble factors from biofilm of single-species MSSA cultures, whereas the soluble factors from biofilm of C. albicans cultures induced only low levels of NO. Soluble factors from 36-h-old biofilm of C. albicans and MSSA cultures promoted cell death and

  17. Two-Dimensional Materials as Prospective Scaffolds for Mixed-Matrix Membrane-Based CO2 Separation.

    PubMed

    Zhu, Xiang; Tian, Chengcheng; Do-Thanh, Chi-Linh; Dai, Sheng

    2017-09-11

    Membrane-based CO 2 separation technology plays a significant role in environmental remediation and clean energy. Two-dimensional (2D) materials with atomically precise structures have emerged as prospective scaffolds to develop mixed-matrix membranes (MMMs) for gas separation. Summarized in this perspective review are the latest breakthrough studies in the synthesis of 2D-material-based MMMs to separate CO 2 from gas mixtures. 2D materials including graphene oxide (GO), metal-organic framework (MOF)-derived nanosheets, covalent organic frameworks (COFs), and transition metal dichalcogenides (TMDs), as fascinating building blocks, have been comprehensively summarized, together with a focus on synthetic processes and gas separation properties. Challenges and the latest advances in the manufacture of novel synthetic 2D materials are briefly discussed to foresee emerging opportunities for the development of new generations of 2D-material-based MMMs. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. A novel pH optical sensor using methyl orange based on triacetylcellulose membranes as support.

    PubMed

    Hosseini, Mohammad; Heydari, Rouhollah; Alimoradi, Mohammad

    2014-07-15

    A novel pH optical sensor based on triacetylcellulose membrane as solid support was developed by using immobilization of methyl orange indicator. The prepared optical sensor was fixed into a flow cell for on-line pH monitoring. Variables affecting sensor performance, such as pH of dye bonding to triacetylcellulose membrane and dye concentration have been fully evaluated and optimized. The calibration curve showed good behavior and precision (RSD<0.4%) in the pH range of 4.0-12.0. No significant variation was observed on sensor response with increasing the ionic strength in the range of 0.0-0.5M of sodium chloride. Determination of pH by using the proposed optical sensor is on-line, quick, inexpensive, selective and sensitive in the pH range of 4.0-12.0. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Engineered catalytic biofilms for continuous large scale production of n-octanol and (S)-styrene oxide.

    PubMed

    Gross, Rainer; Buehler, Katja; Schmid, Andreas

    2013-02-01

    This study evaluates the technical feasibility of biofilm-based biotransformations at an industrial scale by theoretically designing a process employing membrane fiber modules as being used in the chemical industry and compares the respective process parameters to classical stirred-tank studies. To our knowledge, catalytic biofilm processes for fine chemicals production have so far not been reported on a technical scale. As model reactions, we applied the previously studied asymmetric styrene epoxidation employing Pseudomonas sp. strain VLB120ΔC biofilms and the here-described selective alkane hydroxylation. Using the non-heme iron containing alkane hydroxylase system (AlkBGT) from P. putida Gpo1 in the recombinant P. putida PpS81 pBT10 biofilm, we were able to continuously produce 1-octanol from octane with a maximal productivity of 1.3 g L ⁻¹(aq) day⁻¹ in a single tube micro reactor. For a possible industrial application, a cylindrical membrane fiber module packed with 84,000 polypropylene fibers is proposed. Based on the here presented calculations, 59 membrane fiber modules (of 0.9 m diameter and 2 m length) would be feasible to realize a production process of 1,000 tons/year for styrene oxide. Moreover, the product yield on carbon can at least be doubled and over 400-fold less biomass waste would be generated compared to classical stirred-tank reactor processes. For the octanol process, instead, further intensification in biological activity and/or surface membrane enlargement is required to reach production scale. By taking into consideration challenges such as biomass growth control and maintaining a constant biological activity, this study shows that a biofilm process at an industrial scale for the production of fine chemicals is a sustainable alternative in terms of product yield and biomass waste production. Copyright © 2012 Wiley Periodicals, Inc.

  20. Ferromagnetic nanoparticles with peroxidase-like activity enhance the cleavage of biological macromolecules for biofilm elimination

    NASA Astrophysics Data System (ADS)

    GaoCurrent Address: University Of Pennsylvania, School Of Dental Medicine, Philadelphia, Pa 19104, Usa. E.-Mail: Gaoliz@Dental. Upenn. Edu, Lizeng; Giglio, Krista M.; Nelson, Jacquelyn L.; Sondermann, Holger; Travis, Alexander J.

    2014-02-01

    Hydrogen peroxide (H2O2) is a ``green chemical'' that has various cleaning and disinfectant uses, including as an anti-bacterial agent for hygienic and medical treatments. However, its efficacy is limited against biofilm-producing bacteria, because of poor penetration into the protective, organic matrix. Here we show new applications for ferromagnetic nanoparticles (Fe3O4, MNPs) with peroxidase-like activity in potentiating the efficacy of H2O2 in biofilm degradation and prevention. Our data show that MNPs enhanced oxidative cleavage of biofilm components (model nucleic acids, proteins, and oligosaccharides) in the presence of H2O2. When challenged with live, biofilm-producing bacteria, the MNP-H2O2 system efficiently broke down the existing biofilm and prevented new biofilms from forming, killing both planktonic bacteria and those within the biofilm. By enhancing oxidative cleavage of various substrates, the MNP-H2O2 system provides a novel strategy for biofilm elimination, and other applications utilizing oxidative breakdown.Hydrogen peroxide (H2O2) is a ``green chemical'' that has various cleaning and disinfectant uses, including as an anti-bacterial agent for hygienic and medical treatments. However, its efficacy is limited against biofilm-producing bacteria, because of poor penetration into the protective, organic matrix. Here we show new applications for ferromagnetic nanoparticles (Fe3O4, MNPs) with peroxidase-like activity in potentiating the efficacy of H2O2 in biofilm degradation and prevention. Our data show that MNPs enhanced oxidative cleavage of biofilm components (model nucleic acids, proteins, and oligosaccharides) in the presence of H2O2. When challenged with live, biofilm-producing bacteria, the MNP-H2O2 system efficiently broke down the existing biofilm and prevented new biofilms from forming, killing both planktonic bacteria and those within the biofilm. By enhancing oxidative cleavage of various substrates, the MNP-H2O2 system provides a novel

  1. Combined biofouling and scaling in membrane feed channels: a new modeling approach.

    PubMed

    Radu, A I; Bergwerff, L; van Loosdrecht, M C M; Picioreanu, C

    2015-01-01

    A mathematical model was developed for combined fouling due to biofilms and mineral precipitates in membrane feed channels with spacers. Finite element simulation of flow and solute transport in two-dimensional geometries was coupled with a particle-based approach for the development of a composite (cells and crystals) foulant layer. Three fouling scenarios were compared: biofouling only, scaling only and combined fouling. Combined fouling causes a quicker flux decline than the summed flux deterioration when scaling and biofouling act independently. The model results indicate that the presence of biofilms leads to more mineral formation due to: (1) an enhanced degree of saturation for salts next to the membrane and within the biofilm; and (2) more available surface for nucleation to occur. The impact of biofilm in accelerating gypsum precipitation depends on the composition of the feed water (eg the presence of NaCl) and the kinetics of crystal nucleation and growth. Interactions between flow, solute transport and biofilm-induced mineralization are discussed.

  2. Relationships between the antibacterial activity of sodium hypochlorite and treatment time and biofilm age in early Enterococcus faecalis biofilms.

    PubMed

    Chau, N P T; Chung, N H; Jeon, J G

    2015-08-01

    To determine the relationships between the antibacterial activity of NaOCl and treatment time and biofilm age in early Enterococcus faecalis biofilms using a linear fitting procedure. Enterococcus faecalis biofilms were formed on hydroxyapatite discs. To investigate the relationship between the antibacterial activity of NaOCl and biofilm age, 22-, 46-, 70- and 94-h-old biofilms were exposed to NaOCl (0-3%) for 5 min. To investigate the relationship between the antibacterial activity of NaOCl and treatment time, 70-h-old biofilms were exposed to NaOCl (0-3%) for 1, 3, 5 and 7 min. After treatment, colony-forming units (CFUs) were counted. To determine the relationships between these variables, linear fitting was performed. The change in the minimum biofilm eradication concentration (MBEC) of NaOCl followed a linear pattern of biofilm age (R = 0.941, R(2)  = 0.886) or treatment time dependence (R = -0.948, R(2)  = 0.898). Below the MBEC, the fitting lines for bacterial CFU count versus NaOCl concentration (R ≤ -0.973, R(2)  ≥ 0.948) in the 22-, 46-, 70- and 94-h-old biofilms implied that the antibacterial activity of NaOCl decreased as the biofilm age increased. The fitting lines for bacterial CFU count versus NaOCl concentration (R ≤ -0.970, R(2)  ≥ 0.942) in the 1-, 3-, 5- and 7-min treatments implied that the antibacterial activity of NaOCl increased with treatment time. These results suggest that the antibacterial activity of NaOCl against early E. faecalis biofilms in root canals may follow a linear pattern depending on biofilm age or treatment time. © 2014 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  3. Adenovirus, MS2 and PhiX174 interactions with drinking water biofilms developed on PVC, cement and cast iron.

    PubMed

    Helmi, K; Menard-Szczebara, F; Lénès, D; Jacob, P; Jossent, J; Barbot, C; Delabre, K; Arnal, C

    2010-01-01

    Biofilms colonizing pipe surfaces of drinking water distribution systems could provide habitat and shelter for pathogenic viruses present in the water phase. This study aims (i) to develop a method to detect viral particles present in a drinking water biofilm and (ii) to study viral interactions with drinking water biofilms. A pilot scale system was used to develop drinking water biofilms on 3 materials (7 cm(2) discs): PVC, cast iron and cement. Biofilms were inoculated with viral model including MS2, PhiX174 or adenovirus. Five techniques were tested to recover virus from biofilms. The most efficient uses beef extract and glycine at pH = 9. After sonication and centrifugation, the pH of the supernatant is neutralized prior to viral analysis. The calculated recovery rates varied from 29.3 to 74.6% depending on the virus (MS2 or PhiX174) and the material. Applying this protocol, the interactions of virus models (MS2 and adenovirus) with drinking water biofilms were compared. Our results show that adsorption of viruses to biofilms depends on their isoelectric points, the disc material and the hydrodynamic conditions. Applying hydrodynamic conditions similar to those existing in drinking water networks resulted in a viral adsorption corresponding to less than 1% of the initial viral load.

  4. Reversibly pH-responsive polyurethane membranes for on-demand intravaginal drug delivery.

    PubMed

    Kim, Seungil; Chen, Yufei; Ho, Emmanuel A; Liu, Song

    2017-01-01

    To provide better protection for women against sexually transmitted infections, on-demand intravaginal drug delivery was attempted by synthesizing reversibly pH-sensitive polyether-polyurethane copolymers using poly(ethylene glycol) (PEG) and 1,4-bis(2-hydroxyethyl)piperazine (HEP). Chemical structure and thermo-characteristics of the synthesized polyurethanes were confirmed by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), 1 H-nuclear magnetic resonance ( 1 H-NMR), and melting point testing. Membranes were cast by solvent evaporation method using the prepared pH-sensitive polyurethanes. The impact of varying pH on membrane swelling and surface morphology was evaluated via swelling ratio change and scanning electron microscopy (SEM). The prepared pH-responsive membranes showed two times higher swelling ratio at pH 4 than pH 7 and pH-triggered switchable surface morphology change. The anionic anti-inflammatory drug diclofenac sodium (NaDF) was used as a model compound for release studies. The prepared pH-responsive polyurethane membranes allowed continuous NaDF release for 24h and around 20% release of total NaDF within 3h at pH 7 but little-to-no drug release at pH 4.5. NaDF permeation across the prepared membranes demonstrated a reversible pH-responsiveness. The pH-responsive polyurethane membranes did not show any noticeable negative impact on vaginal epithelial cell viability or induction of pro-inflammatory cytokine production compared to controls. Overall, the non-cytotoxic HEP-based pH-responsive polyurethane demonstrated its potential to be used in membrane-based implants such as intravaginal rings to achieve on-demand "on-and-off" intravaginal drug delivery. A reversible and sharp switch between "off" and "on" drug release is achieved for the first time through new pH-sensitive polyurethane membranes, which can serve as window membranes in reservoir-type intravaginal rings for on-demand drug delivery to prevent sexually

  5. Susceptibility of Staphylococcus aureus biofilms to reactive discharge gases.

    PubMed

    Traba, Christian; Liang, Jun F

    2011-08-01

    Formation of bacterial biofilms at solid-liquid interfaces creates numerous problems in both industrial and biomedical sciences. In this study, the susceptibility of Staphylococcus aureus biofilms to discharge gas generated from plasma was tested. It was found that despite distinct chemical/physical properties, discharge gases from oxygen, nitrogen, and argon demonstrated very potent and almost the same anti-biofilm activity. The bacterial cells in S. aureus biofilms were killed (>99.9%) by discharge gas within minutes of exposure. Under optimal experimental conditions, no bacteria and biofilm re-growth from discharge gas treated biofilms was found. Further studies revealed that the anti-biofilm activity of the discharge gas occurred by two distinct mechanisms: (1) killing bacteria in biofilms by causing severe cell membrane damage, and (2) damaging the extracellular polymeric matrix in the architecture of the biofilm to release biofilm from the surface of the solid substratum. Information gathered from this study provides an insight into the anti-biofilm mechanisms of plasma and confirms the applications of discharge gas in the treatment of biofilms and biofilm related bacterial infections.

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

    PubMed

    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.

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

    PubMed Central

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

    2015-01-01

    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. PMID:26272750

  8. Insights into the distribution of water in a self-humidifying H2/O2 proton-exchange membrane fuel cell using 1H NMR microscopy.

    PubMed

    Feindel, Kirk W; Bergens, Steven H; Wasylishen, Roderick E

    2006-11-01

    Proton ((1)H) NMR microscopy is used to investigate in-situ the distribution of water throughout a self-humidifying proton-exchange membrane fuel cell, PEMFC, operating at ambient temperature and pressure on dry H(2)(g) and O(2)(g). The results provide the first experimental images of the in-plane distribution of water within the PEM of a membrane electrode assembly in an operating fuel cell. The effect of gas flow configuration on the distribution of water in the PEM and cathode flow field is investigated, revealing that the counter-flow configurations yield a more uniform distribution of water throughout the PEM. The maximum power output from the PEMFC, while operating under conditions of constant external load, occurs when H(2)O(l) is first visible in the (1)H NMR image of the cathode flow field, and subsequently declines as this H(2)O(l) continues to accumulate. The (1)H NMR microscopy experiments are in qualitative agreement with predictions from several theoretical modeling studies (e.g., Pasaogullari, U.; Wang, C. Y. J. Electrochem. Soc. 2005, 152, A380-A390), suggesting that combined theoretical and experimental approaches will constitute a powerful tool for PEMFC design, diagnosis, and optimization.

  9. Engineered chimeric peptides with antimicrobial and titanium-binding functions to inhibit biofilm formation on Ti implants.

    PubMed

    Geng, Hongjuan; Yuan, Yang; Adayi, Aidina; Zhang, Xu; Song, Xin; Gong, Lei; Zhang, Xi; Gao, Ping

    2018-01-01

    Titanium (Ti) implants have been commonly used in oral medicine. However, despite their widespread clinical application, these implants are susceptible to failure induced by microbial infection due to bacterial biofilm formation. Immobilization of chimeric peptides with antibacterial properties on the Ti surface may be a promising antimicrobial approach to inhibit biofilm formation. Here, chimeric peptides were designed by connecting three sequences (hBD-3-1/2/3) derived from human β-defensin-3 (hBD-3) with Ti-binding peptide-l (TBP-l: RKLPDAGPMHTW) via a triple glycine (G) linker to modify Ti surfaces. Using X-ray photoelectron spectroscopy (XPS), the properties of individual domains of the chimeric peptides were evaluated for their binding activity toward the Ti surface. The antimicrobial and anti-biofilm efficacy of the peptides against initial settlers, Streptococcus oralis (S. oralis), Streptococcus gordonii (S. gordonii) and Streptococcus sanguinis (S. sanguinis), was evaluated with confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Transmission electron microscopy (TEM) and real-time quantitative PCR (qRT-PCR) were used to study cell membrane changes and the underlying antimicrobial mechanism. Compared with the other two peptides, TBP-1-GGG-hBD3-3 presented stronger antibacterial activity and remained stable in saliva and serum. Therefore, it was chosen as the best candidate to modify Ti surfaces in this study. This peptide inhibited the growth of initial streptococci and biofilm formation on Ti surfaces with no cytotoxicity to MC3T3-E1 cells. Disruption of the integrity of bacterial membranes and decreased expression of adhesion protein genes from S. gordonii revealed aspects of the antibacterial mechanism of TBP-1-GGG-hBD3-3. We conclude that engineered chimeric peptides with antimicrobial activity provide a potential solution for inhibiting biofilm formation on Ti surfaces to reduce or prevent the occurrence of peri

  10. Biofilm and planktonic lifestyles differently support the resistance of the desert cyanobacterium Chroococcidiopsis under space and Martian simulations.

    PubMed

    Baqué, Mickael; Scalzi, Giuliano; Rabbow, Elke; Rettberg, Petra; Billi, Daniela

    2013-10-01

    When Chroococcidiopsis sp. strain CCMEE 057 from the Sinai Desert and strain CCMEE 029 from the Negev Desert were exposed to space and Martian simulations in the dried status as biofilms or multilayered planktonic samples, the biofilms exhibited an enhanced rate of survival. Compared to strain CCMEE 029, biofilms of strain CCME 057 better tolerated UV polychromatic radiation (5 × 10(5) kJ/m(2) attenuated with a 0.1% neutral density filter) combined with space vacuum or Martian atmosphere of 780 Pa. CCMEE 029, on the other hand, failed to survive UV polychromatic doses higher than 1.5 × 10(3) kJ/m(2). The induced damage to genomic DNA, plasma membranes and photosynthetic apparatus was quantified and visualized by means of PCR-based assays and CLSM imaging. Planktonic samples of both strains accumulated a higher amount of damage than did the biofilms after exposure to each simulation; CLSM imaging showed that photosynthetic pigment bleaching, DNA fragmentation and damaged plasma membranes occurred in the top 3-4 cell layers of both biofilms and of multilayered planktonic samples. Differences in the EPS composition were revealed by molecular probe staining as contributing to the enhanced endurance of biofilms compared to that of planktonic samples. Our results suggest that compared to strain CCMEE 029, biofilms of strain CCMEE 057 might better tolerate 1 year's exposure in space during the next EXPOSE-R2 mission.

  11. Biofilm and Planktonic Lifestyles Differently Support the Resistance of the Desert Cyanobacterium Chroococcidiopsis Under Space and Martian Simulations

    NASA Astrophysics Data System (ADS)

    Baqué, Mickael; Scalzi, Giuliano; Rabbow, Elke; Rettberg, Petra; Billi, Daniela

    2013-10-01

    When Chroococcidiopsis sp. strain CCMEE 057 from the Sinai Desert and strain CCMEE 029 from the Negev Desert were exposed to space and Martian simulations in the dried status as biofilms or multilayered planktonic samples, the biofilms exhibited an enhanced rate of survival. Compared to strain CCMEE 029, biofilms of strain CCME 057 better tolerated UV polychromatic radiation (5 × 105 kJ/m2 attenuated with a 0.1 % neutral density filter) combined with space vacuum or Martian atmosphere of 780 Pa. CCMEE 029, on the other hand, failed to survive UV polychromatic doses higher than 1.5 × 103 kJ/m2. The induced damage to genomic DNA, plasma membranes and photosynthetic apparatus was quantified and visualized by means of PCR-based assays and CLSM imaging. Planktonic samples of both strains accumulated a higher amount of damage than did the biofilms after exposure to each simulation; CLSM imaging showed that photosynthetic pigment bleaching, DNA fragmentation and damaged plasma membranes occurred in the top 3-4 cell layers of both biofilms and of multilayered planktonic samples. Differences in the EPS composition were revealed by molecular probe staining as contributing to the enhanced endurance of biofilms compared to that of planktonic samples. Our results suggest that compared to strain CCMEE 029, biofilms of strain CCMEE 057 might better tolerate 1 year's exposure in space during the next EXPOSE-R2 mission.

  12. Method for forming H2-permselective oxide membranes

    DOEpatents

    Gavalas, G.R.; Nam, S.W.; Tsapatsis, M.; Kim, S.

    1995-09-26

    Methods are disclosed for forming permselective oxide membranes that are highly selective to permeation of hydrogen by chemical deposition of reactants in the pore of porous tubes, such as Vycor{trademark} glass or Al{sub 2}O{sub 3} tubes. The porous tubes have pores extending through the tube wall. The process involves forming a stream containing a first reactant of the formula RX{sub n}, wherein R is silicon, titanium, boron or aluminum, X is chlorine, bromine or iodine, and n is a number which is equal to the valence of R; and forming another stream containing water vapor as the second reactant. Both of the reactant streams are passed along either the outside or the inside surface of a porous tube and the streams react in the pores of the porous tube to form a nonporous layer of R-oxide in the pores. The membranes are formed by the hydrolysis of the respective halides. In another embodiment, the first reactant stream contains a first reactant having the formula SiH{sub n}Cl{sub 4{minus}n} where n is 1, 2 or 3; and the second reactant stream contains water vapor and oxygen. In still another embodiment the first reactant stream containing a first reactant selected from the group consisting of Cl{sub 3}SiOSiCl{sub 3}, Cl{sub 3}SiOSiCl{sub 2}OSiCl{sub 3}, and mixtures thereof and the second reactant stream contains water vapor. In still another embodiment, membrane formation is carried out by an alternating flow deposition method. This involves a sequence of cycles, each cycle comprising introduction of the halide-containing stream and allowance of a specific time for reaction followed by purge and flow of the water vapor containing stream for a specific length of time. In all embodiments the nonporous layers formed are selectively permeable to hydrogen. 11 figs.

  13. Comparative in vitro efficacies of ethanol-, EDTA- and levofloxacin-based catheter lock solutions on eradication of Stenotrophomonas maltophilia biofilms.

    PubMed

    Passerini de Rossi, Beatriz; Feldman, Laureana; Pineda, María Saliba; Vay, Carlos; Franco, Mirta

    2012-09-01

    The aim of this study was to compare the in vitro activity of ethanol, EDTA and levofloxacin (Levo), alone or in combination, on biofilms of Stenotrophomonas maltophilia recovered from patients with catheter-related bloodstream infections (CRBSIs) at a university hospital in Argentina. First, 24 and 48 h biofilms were formed in microtitre plates and challenged with 25 or 40 % ethanol for 1 h. Biofilms, of the 14 local isolates and from the reference strain K279a, were eradicated after both treatments as shown by plate counts and the regrowth technique. Second, 24 h biofilms of all isolates were established in silicone catheter segments and challenged with 25 or 40 % ethanol, Levo (2.5 mg ml(-1)), EDTA (30 mg ml(-1)), 25 % ethanol-EDTA or Levo-EDTA for 1, 3 and 24 h. Viable counts of biofilms treated for 1 h with 25 or 40 % ethanol or 25 % ethanol-EDTA were under the limit of detection. Killing of biofilms by Levo or Levo-EDTA was gradual and it was only after 24 h of treatment that no differences could be seen between the effects of these catheter lock solutions (CLSs) and those of ethanol (P>0.05). Levo-EDTA, in combination, did not act synergistically against biofilms. After 24 h of exposure, EDTA did not eradicate biofilms but reduced biofilm survival rates to 1-5 %. The effect of the different CLSs on biomass reduction, estimated by crystal violet staining, was highly dependent on the isolate, and the most effective agents were 25 and 40 % ethanol. Our results suggest that when used as a CLS for short periods, ethanol at low concentrations, alone or in combination with a chelator, can decontaminate the line from S. maltophilia in cases of CRBSI and help, in conjunction with systemic antibiotics, in the retention of precious vascular catheters.

  14. Immobilization of Rhodococcus rhodochrous BX2 (an acetonitrile-degrading bacterium) with biofilm-forming bacteria for wastewater treatment.

    PubMed

    Li, Chunyan; Li, Yue; Cheng, Xiaosong; Feng, Liping; Xi, Chuanwu; Zhang, Ying

    2013-03-01

    In this study, a unique biofilm consisting of three bacterial strains with high biofilm-forming capability (Bacillus subtilis E2, E3, and N4) and an acetonitrile-degrading bacterium (Rhodococcus rhodochrous BX2) was established for acetonitrile-containing wastewater treatment. The results indicated that this biofilm exhibited strong resistance to acetonitrile loading shock and displayed a typical spatial and structural heterogeneity and completely depleted the initial concentration of acetonitrile (800mgL(-1)) within 24h in a moving-bed-biofilm reactor (MBBR) after operation for 30days. The immobilization of BX2 cells in the biofilm was confirmed by PCR-DGGE. It has been demonstrated that biofilm-forming bacteria can promote the immobilization of contaminant-degrading bacteria in the biofilms and can subsequently improve the degradation of contaminants in wastewater. This approach offers a novel strategy for enhancing biological oxidation of toxic pollutants in wastewater. Copyright © 2012 Elsevier Ltd. All rights reserved.

  15. Biofilm Formation by Helicobacter pylori and Its Involvement for Antibiotic Resistance

    PubMed Central

    Yonezawa, Hideo; Osaki, Takako

    2015-01-01

    Bacterial biofilms are communities of microorganisms attached to a surface. Biofilm formation is critical not only for environmental survival but also for successful infection. Helicobacter pylori is one of the most common causes of bacterial infection in humans. Some studies demonstrated that this microorganism has biofilm forming ability in the environment and on human gastric mucosa epithelium as well as on in vitro abiotic surfaces. In the environment, H. pylori could be embedded in drinking water biofilms through water distribution system in developed and developing countries so that the drinking water may serve as a reservoir for H. pylori infection. In the human stomach, H. pylori forms biofilms on the surface of gastric mucosa, suggesting one possible explanation for eradication therapy failure. Finally, based on the results of in vitro analyses, H. pylori biofilm formation can decrease susceptibility to antibiotics and H. pylori antibiotic resistance mutations are more frequently generated in biofilms than in planktonic cells. These observations indicated that H. pylori biofilm formation may play an important role in preventing and controlling H. pylori infections. Therefore, investigation of H. pylori biofilm formation could be effective in elucidating the detailed mechanisms of infection and colonization by this microorganism. PMID:26078970

  16. Extracellular DNA in Helicobacter pylori biofilm: a backstairs rumour.

    PubMed

    Grande, R; Di Giulio, M; Bessa, L J; Di Campli, E; Baffoni, M; Guarnieri, S; Cellini, L

    2011-02-01

    This study detected and characterized the extracellular DNA (eDNA) in the biofilm extracellular polymeric substance (EPS) matrix of Helicobacter pylori and investigated the role of such component in the biofilm development. Extracellular DNA was purified and characterized in a 2-day-old mature biofilm developed by the reference strain H. pylori ATCC 43629, the clinical isolate H. pylori SDB60 and the environmental strain H. pylori MDC1. Subsequently, the role of eDNA in the H. pylori biofilm was evaluated by adding DNase I during biofilm formation and on mature biofilms. Extracellular DNA was detected in the 2-day-old EPS biofilm matrix of all analysed H. pylori strains. The DNA fingerprintings, performed by RAPD analysis, on eDNA and intracellular DNA (iDNA), showed some remarkable differences. The data obtained by microtitre biofilm assay as well as colony forming unit count and CLSM (confocal laser scanning microscopy) qualitative analysis did not show any significant differences between the DNase I-treated biofilms and the corresponding not treated controls both in formation and on mature biofilms. In this study, we provide evidence that eDNA is a component of the EPS matrix of H. pylori biofilm. The different profiles of eDNA and iDNA indicate that lysed cells are not the primary source of eDNA release, suggesting that other active mechanisms might be involved in this process. Moreover, the biomass assay suggests that eDNA may not be the main component of biofilm matrix, suggesting that it could be primarily involved in other mechanisms such as recombination processes, via transformation, contributing to the wide genomic variability of this micro-organism defined as a 'quasi-species'. The presence of eDNA in H. pylori biofilm can contribute to the active dynamic exchange of information aimed to reach the best condition for the bacterial survival in the host and in the environment. © 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for

  17. Removal of Burkholderia cepacia biofilms with oxidants

    NASA Technical Reports Server (NTRS)

    Koenig, D. W.; Mishra, S. K.; Pierson, D. L.

    1995-01-01

    Iodine is used to disinfect the water system aboard US space shuttles and is the anticipated biocide for the international space station. Water quality on spacecraft must be maintained at the highest possible levels for the safety of the crew. Furthermore, the treatment process used to maintain the quality of water on research must be robust and operate for long periods with minimal crew intervention. Biofilms are recalcitrant and pose a major threat with regard to chronic contamination of spacecraft water systems. We measured the effectiveness of oxidizing biocides on the removal and regrowth of Burkholderia (Pseudomonas) cepacia biofilms. B. cepacia, isolated from the water distribution system of the space shuttle Discovery, was grown in continuous culture to produce a bacterial contamination source for biofilm formation and removal studies. A 10(7) CFU ml-1 B. cepacia suspension, in distilled water, was used to form biofilms on 3000 micrometers2 glass surfaces. Rates of attachment were measured directly with image analysis and were found to be 7.8, 15.2, and 22.8 attachment events h-1 for flow rates of 20.7, 15.2, and 9.8 ml min-1, respectively. After 18 h of formation, the B. cepacia biofilms were challenged with oxidants (ozone, chlorine, and iodine) and the rates of biofilm removal determined by image analysis. Fifty percent of the biofilm material was removed in the first hour of continous treatment with 24 mg l-1 chlorine or 2 mg l-1 ozone. Iodine (48 mg l-1) did not remove any measurable cellular material after 6 h continuous contact. After this first removal of biofilms by the oxidants, the surface was allowed to refoul and was again treated with the biocide. Iodine was the only compound that was unable to remove cellular debris from either primary or secondary biofilms. Moreover, treating primary biofilms with iodine increased the rate of formation of secondary biofilms, from 4.4 to 5.8 attachment events h-1. All the oxidants tested inactivated the B

  18. Role of beta 1-4 linked polymers in the biofilm structure of marine Pseudomonas sp. CE-2 on 304 stainless steel coupons.

    PubMed

    Jain, Anand; Bhosle, Narayan B

    2008-01-01

    Pseudomonas sp CE-2 cells attach and form biofilms on 304-stainless steel (SS) coupons. A series of experiments were carried out in order to understand the role of exopolysaccharides (EPS) in the formation and maintenance of CE-2 biofilms on SS coupons. The biofilm density and EPS concentration increased over the period of incubation and the highest values for both were recorded after 72 h. Calcofluor and the lectin concanavalin A (Con A) showed a positive interaction with 72-h old biofilms, indicating the presence of beta 1-4 linked polymers, and alpha-d-glucose and alpha-d-mannose in the biofilm matrix of CE-2. When the CE-2 cells were grown in the presence of calcofluor (200 microg ml(-1)), biofilm formation was significantly reduced (approximately 85%). Conversely, the lectins Con A or WGA did not influence the CE-2 biofilms on the SS coupons. Furthermore, treatment with cellulase, an enzyme specific for the degradation of beta 1-4 linked polymers, removed substantial amounts of CE-2 biofilm from SS coupons. These results strongly suggest the involvement of beta 1-4 linked polymers in the formation and maintenance of Pseudomonas sp. CE-2 biofilms on SS coupons.

  19. Mechanisms of nitrous oxide (N2 O) formation and reduction in denitrifying biofilms.

    PubMed

    Sabba, Fabrizio; Picioreanu, Cristian; Nerenberg, Robert

    2017-12-01

    Nitrous oxide (N 2 O) is a potent greenhouse gas that can be formed in wastewater treatment processes by ammonium oxidizing and denitrifying microorganisms. While N 2 O emissions from suspended growth systems have been extensively studied, and some recent studies have addressed emissions from nitrifying biofilms, much less is known about N 2 O emissions from denitrifying biofilm processes. This research used modeling to evaluate the mechanisms of N 2 O formation and reduction in denitrifying biofilms. The kinetic model included formation and consumption of key denitrification species, including nitrate (NO3-), nitrite (NO2-), nitric oxide (NO), and N 2 O. The model showed that, in presence of excess of electron donor, denitrifying biofilms have two distinct layers of activity: an outer layer where there is net production of N 2 O and an inner layer where there is net consumption. The presence of oxygen (O 2 ) had an important effect on N 2 O emission from suspended growth systems, but a smaller effect on biofilm systems. The effects of NO3- and O 2 differed significantly based on the biofilm thickness. Overall, the effects of biofilm thickness and bulk substrate concentrations on N 2 O emissions are complex and not always intuitive. A key mechanism for denitrifying biofilms is the diffusion of N 2 O and other intermediates from one zone of the biofilm to another. This leads to zones of N 2 O formation or consumption transformations that would not exist in suspended growth systems. © 2017 Wiley Periodicals, Inc.

  20. Membrane Interactions of hIAPP Monomer and Oligomer with Lipid Membranes by Molecular Dynamics Simulations.

    PubMed

    Zhang, Mingzhen; Ren, Baiping; Liu, Yonglan; Liang, Guizhao; Sun, Yan; Xu, Lijian; Zheng, Jie

    2017-08-16

    Interaction of human islet amyloid polypeptide (hIAPP) peptides with cell membrane is crucial for the understanding of amyloid toxicity associated with Type II diabetes (T2D). While it is known that the hIAPP-membrane interactions are considered to promote hIAPP aggregation into fibrils and induce membrane disruption, the membrane-induced conformation, orientation, aggregation, and adsorption behaviors of hIAPP peptides have not been well understood at the atomic level. Herein, we perform all-atom explicit-water molecular dynamics (MD) simulations to study the adsorption, orientation, and surface interaction of hIAPP aggregates with different sizes (monomer to tetramer) and conformations (monomer with α-helix and tetramer with β-sheet-rich U-turn) upon adsorption on the lipid bilayers composed of both pure zwitterionic POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and mixed anionic POPC/POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) (3:1) lipids. MD simulation results show that hIAPP monomer with α-helical conformation and hIAPP pentamer with β-sheet conformation can adsorb on both POPC and POPC/POPE bilayers via a preferential orientation of N-terminal residues facing toward the bilayer surface. The hIAPP aggregates show stronger interactions with mixed POPC/POPE lipids than pure POPC lipids, consistent with experimental observation that hIAPP adsorption and fibrililation are enhanced on mixed lipid bilayers. While electrostatic interactions are main attractive forces to drive the hIAPP aggregates to adsorb on both bilayers, the introduction of the more hydrophilic head groups of POPE lipids further promote the formation of the interfacial hydrogen bonds. Complement to our previous studies of hIAPP aggregates in bulk solution, this computational work increases our knowledge about the mechanism of amyloid peptide-membrane interactions that is central to the understanding the progression of all amyloid diseases.

  1. Inhibition of Pseudomonas aeruginosa biofilm formation by 2,2'-bipyridyl, lipoic, kojic and picolinic acids.

    PubMed

    Çevik, Kübra; Ulusoy, Seyhan

    2015-08-01

    The inhibitory effects of iron chelators, and FeCl3 chelation on biofilm formation and swarming motility were investigated against an opportunistic human pathogen Pseudomonas aeruginosa. The inhibitory activity of 2,2'-bipyridyl, lipoic acid, kojic acid and picolinic acid on biofilm formation of P. aeruginosa strain PAO1 and three clinical isolates (P. aeruginosa PAK01, P. aeruginosa PAK02 and P. aeruginosa PAK03) were investigated, based on crystal violet assay, and swarming motility test. The kojic, lipoic and picolinic acid inhibited biofilm formation by 5-33% in all tested P. aeruginosa isolates. When chelated iron was added, biofilm inhibition rates were determined to be 39-57%. Among the tested chelators against P. aeruginosa, lipoic acid (84%) and kojic acid (68%) presented the highest inhibition of swarming motility. This is the first study to report the inhibitory effect of lipoic acid on biofilm formation and swarming motility of P. aeruginosa. It is considered that lipoic and picolinic acids can serve as alternatives for the treatment of the P. aeruginosa infections by inhibiting biofilm formation.

  2. Essential Oils and Eugenols Inhibit Biofilm Formation and the Virulence of Escherichia coli O157:H7

    PubMed Central

    Kim, Yong-Guy; Lee, Jin-Hyung; Gwon, Giyeon; Kim, Soon-Il; Park, Jae Gyu; Lee, Jintae

    2016-01-01

    Enterohemorrhagic Escherichia coli O157:H7 (EHEC) has caused foodborne outbreaks worldwide and the bacterium forms antimicrobial-tolerant biofilms. We investigated the abilities of various plant essential oils and their components to inhibit biofilm formation by EHEC. Bay, clove, pimento berry oils and their major common constituent eugenol at 0.005% (v/v) were found to markedly inhibit EHEC biofilm formation without affecting planktonic cell growth. In addition, three other eugenol derivatives isoeugenol, 2-methoxy-4-propylphenol, and 4-ethylguaiacol had antibiofilm activity, indicating that the C-1 hydroxyl unit, the C-2 methoxy unit, and C-4 alkyl or alkane chain on the benzene ring of eugenol play important roles in antibiofilm activity. Interestingly, these essential oils and eugenol did not inhibit biofilm formation by three laboratory E. coli K-12 strains that reduced curli fimbriae production. Transcriptional analysis showed that eugenol down-regulated 17 of 28 genes analysed, including curli genes (csgABDFG), type I fimbriae genes (fimCDH) and ler-controlled toxin genes (espD, escJ, escR, and tir), which are required for biofilm formation and the attachment and effacement phenotype. In addition, biocompatible poly(lactic-co-glycolic acid) coatings containing clove oil or eugenol exhibited efficient biofilm inhibition on solid surfaces. In a Caenorhabditis elegans nematode model, clove oil and eugenol attenuated the virulence of EHEC. PMID:27808174

  3. Deoxycholate-Based Glycosides (DCGs) for Membrane Protein Stabilisation.

    PubMed

    Bae, Hyoung Eun; Gotfryd, Kamil; Thomas, Jennifer; Hussain, Hazrat; Ehsan, Muhammad; Go, Juyeon; Loland, Claus J; Byrne, Bernadette; Chae, Pil Seok

    2015-07-06

    Detergents are an absolute requirement for studying the structure of membrane proteins. However, many conventional detergents fail to stabilise denaturation-sensitive membrane proteins, such as eukaryotic proteins and membrane protein complexes. New amphipathic agents with enhanced efficacy in stabilising membrane proteins will be helpful in overcoming the barriers to studying membrane protein structures. We have prepared a number of deoxycholate-based amphiphiles with carbohydrate head groups, designated deoxycholate-based glycosides (DCGs). These DCGs are the hydrophilic variants of previously reported deoxycholate-based N-oxides (DCAOs). Membrane proteins in these agents, particularly the branched diglucoside-bearing amphiphiles DCG-1 and DCG-2, displayed favourable behaviour compared to previously reported parent compounds (DCAOs) and conventional detergents (LDAO and DDM). Given their excellent properties, these agents should have significant potential for membrane protein studies. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Transfer, attachment, and formation of biofilms by Escherichia coli O157:H7 on meat-contact surface materials.

    PubMed

    Simpson Beauchamp, Catherine; Dourou, Dimitra; Geornaras, Ifigenia; Yoon, Yohan; Scanga, John A; Belk, Keith E; Smith, Gary C; Nychas, George-John E; Sofos, John N

    2012-06-01

    Studies examined the effects of meat-contact material types, inoculation substrate, presence of air at the liquid-solid surface interface during incubation, and incubation substrate on the attachment/transfer and subsequent biofilm formation by Escherichia coli O157:H7 on beef carcass fabrication surface materials. Materials studied as 2 × 5 cm coupons included stainless steel, acetal, polypropylene, and high-density polyethylene. A 6-strain rifampicin-resistant E. coli O157:H7 composite was used to inoculate (6 log CFU/mL, g, or cm²) tryptic soy broth (TSB), beef fat/lean tissue homogenate (FLH), conveyor belt-runoff fluids, ground beef, or beef fat. Coupons of each material were submerged (4 °C, 30 min) in the inoculated fluids or ground beef, or placed between 2 pieces of inoculated beef fat with pressure (20 kg) applied. Attachment/transfer of the pathogen was surface material and substrate dependent, although beef fat appeared to negate differences among surface materials. Beef fat was the most effective (P < 0.05) inoculation substrate, followed by ground beef, FLH, and TSB. Incubation (15 °C, 16 d) of beef fat-inoculated coupons in a beef fat homogenate (pH 4.21) allowed the pathogen to survive and grow on coupon surfaces, with maximal biofilm formation observed between 2 and 8 d of storage and when air was present at the liquid-solid interface. The results indicated that the process of fabricating beef carcasses may be conducive to the attachment of E. coli O157:H7 onto meat-contact surfaces and subsequent biofilm formation. Furthermore, it is recommended that substrates found in beef fabrication settings, rather than laboratory culture media, be used in studies designed to investigate E. coli O157:H7 biofilm development and control in these environments. Findings of this study provide knowledge on the effect of type of beef carcass fabrication surface material, fabrication-floor fluids and residues, and incubation conditions on attachment/transfer and

  5. Behavior of nitrogen removal in an aerobic sponge based moving bed biofilm reactor.

    PubMed

    Zhang, Xinbo; Song, Zi; Guo, Wenshan; Lu, Yanmin; Qi, Li; Wen, Haitao; Ngo, Huu Hao

    2017-12-01

    This study aims to investigate the behavior of nitrogen removal in an aerobic sponge based moving bed biofilm reactor by evaluating nitrification and denitrification rates of sponge biocarriers from three aerobic moving bed biofilm reactors (MBBRs) with filling ratios of 10% (R-10), 20% (R-20) and 30% (R-30). Results showed that the highest removal efficiencies of total nitrogen in three reactors were 84.5% (R-10), 93.6% (R-20) and 95.3% (R-30). Correspondingly, simultaneous nitrification and denitrification rate (SND) was 90.9%, 97.6% and 100%, respectively. Although R-20 had the highest attached-growth biomass (AGB) per gram of sponge compared to the other two reactors, R-30 showed the maximum ammonium oxidation rate (AOR) (2.1826±0.0717mgNH 4 + -N/gAGB/h) and denitrification rate (DNR) (5.0852±0.0891mgNO 3 - -N/gAGB/h), followed by R-20 and R-10. These results indicated AOR, DNR and AGB were affected by the filling ratio under the same operation mode. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. A study on the interactions of Aurein 2.5 with bacterial membranes.

    PubMed

    Dennison, Sarah R; Morton, Leslie H G; Shorrocks, Andrea J; Harris, Frederick; Phoenix, David A

    2009-02-01

    Aurein 2.5 (GLFDIVKKVVGAFGSL-NH(2)) is an uncharacterised antimicrobial peptide. At an air/water interface, it exhibited strong surface activity (maximal surface pressure 25mNm(-1)) and molecular areas consistent with the adoption of alpha-helical structure orientated either perpendicular (1.72nm(2)molecule(-1)) or parallel (3.6nm(2)molecule(-1)) to the interface. Aurein 2.5 was strongly antibacterial, exhibiting a minimum inhibitory concentration (MIC) of 30microM against Bacillus subtilis and Escherichia coli. The peptide induced maximal surface pressure changes of 9mNm(-1) and 5mNm(-1), respectively, in monolayers mimicking membranes of these organisms whilst compression isotherm analysis of these monolayers showed DeltaG(Mix)>0, indicating destabilisation by Aurein 2.5. These combined data suggested that toxicity of the peptide to these organisms may involve membrane invasion via the use of oblique orientated alpha-helical structure. The peptide induced strong, comparable maximal surface changes in monolayers of DOPG (7.5mNm(-1)) and DOPE monolayers (6mNm(-1)) suggesting that the membrane interactions of Aurein 2.5 were driven by amphiphilicity rather than electrostatic interaction. Based on these data, it was suggested that the differing ability of Aurein 2.5 to insert into membranes of B. subtilis and E. coli was probably related to membrane-based factors such as differences in lipid packing characteristics. The peptide was active against both sessile E. coli and Staphylococcus aureus with an MIC of 125microM. The broad-spectrum antibacterial activity and non-specific modes of membrane action used by Aurein 2.5 suggested use as an anti-biofilm agent such as in the decontamination of medical devices.

  7. Effects of plastic composite support and pH profiles on pullulan production in a biofilm reactor.

    PubMed

    Cheng, Kuan-Chen; Demirci, Ali; Catchmark, Jeffrey M

    2010-04-01

    Pullulan is a linear homopolysaccharide which is composed of glucose units and often described as alpha-1, 6-linked maltotriose. The applications of pullulan range from usage as blood plasma substitutes to environmental pollution control agents. In this study, a biofilm reactor with plastic composite support (PCS) was evaluated for pullulan production using Aureobasidium pullulans. In test tube fermentations, PCS with soybean hulls, defatted soy bean flour, yeast extract, dried bovine red blood cells, and mineral salts was selected for biofilm reactor fermentation (due to its high nitrogen content, moderate nitrogen leaching rate, and high biomass attachment). Three pH profiles were later applied to evaluate their effects on pullulan production in a PCS biofilm reactor. The results demonstrated that when a constant pH at 5.0 was applied, the time course of pullulan production was advanced and the concentration of pullulan reached 32.9 g/L after 7-day cultivation, which is 1.8-fold higher than its respective suspension culture. The quality analysis demonstrated that the purity of produced pullulan was 95.8% and its viscosity was 2.4 centipoise. Fourier transform infrared spectroscopy spectra also supported the supposition that the produced exopolysaccharide was mostly pullulan. Overall, this study demonstrated that a biofilm reactor can be successfully implemented to enhance pullulan production and maintain its high purity.

  8. Effect of Electrolyzed Water on the Disinfection of Bacillus cereus Biofilms: The Mechanism of Enhanced Resistance of Sessile Cells in the Biofilm Matrix.

    PubMed

    Hussain, Mohammad Shakhawat; Kwon, Minyeong; Tango, Charles Nkufi; Oh, Deog Hwan

    2018-05-01

    This study examined the disinfection efficacy and mechanism of electrolyzed water (EW) on Bacillus cereus biofilms. B. cereus strains, ATCC 14579 and Korean Collection for Type Cultures (KCTC) 13153 biofilms, were formed on stainless steel (SS) and plastic slide (PS) coupons. Mature biofilms were treated with slightly acidic EW (SAEW), acidic EW (AEW), and basic EW (BEW). SAEW (available chlorine concentration, 25 ± 1.31 mg L -1 ; pH 5.71 ± 0.16; and oxidation reduction potential, 818 to 855 mV) reduced ATCC 14579 biofilms on plastic slides to below the detection limit within 30 s. However, biofilms on SS coupons showed a higher resistance to the SAEW treatment. When the disinfection activities of three types of EW on biofilms were compared, AEW showed a higher bactericidal activity, followed by SAEW and BEW. In contrast, BEW showed a significantly ( P < 0.05) higher biofilm dispersal activity than AEW and SAEW. SAEW disinfection of the B. cereus biofilms was due to the disruption of the B. cereus plasma membrane. The higher resistance of biofilms formed on the SS coupon might be due to the higher number of attached cells and extracellular polymeric substances formation that reacts with the active chlorine ions, such as hypochlorous acid and hypochlorite ion of SAEW, which decreased the disinfection efficacy of SAEW. This study showed that the EW treatment effectively disinfected B. cereus biofilms, providing insight into the potential use of EW in the food processing industry to control the biofilm formation of B. cereus.

  9. Extracellular DNA Contributes to Dental Biofilm Stability.

    PubMed

    Schlafer, Sebastian; Meyer, Rikke L; Dige, Irene; Regina, Viduthalai R

    2017-01-01

    Extracellular DNA (eDNA) is a major matrix component of many bacterial biofilms. While the presence of eDNA and its role in biofilm stability have been demonstrated for several laboratory biofilms of oral bacteria, there is no data available on the presence and function of eDNA in in vivo grown dental biofilms. This study aimed to determine whether eDNA was part of the matrix in biofilms grown in situ in the absence of sucrose and whether treatment with DNase dispersed biofilms grown for 2.5, 5, 7.5, 16.5, or 24 h. Three hundred biofilms from 10 study participants were collected and treated with either DNase or heat-inactivated DNase for 1 h. The bacterial biovolume was determined with digital image analysis. Staining with TOTO®-1 allowed visualization of eDNA both on bacterial cell surfaces and, with a cloud-like appearance, in the intercellular space. DNase treatment strongly reduced the amount of biofilm in very early stages of growth (up to 7.5 h), but the treatment effect decreased with increasing biofilm age. This study proves the involvement of eDNA in dental biofilm formation and its importance for biofilm stability in the earliest stages. Further research is required to uncover the interplay of eDNA and other matrix components and to explore the therapeutic potential of DNase treatment for biofilm control. © 2017 S. Karger AG, Basel.

  10. A Dual-Phase Ceramic Membrane with Extremely High H2 Permeation Flux Prepared by Autoseparation of a Ceramic Precursor.

    PubMed

    Cheng, Shunfan; Wang, Yanjie; Zhuang, Libin; Xue, Jian; Wei, Yanying; Feldhoff, Armin; Caro, Jürgen; Wang, Haihui

    2016-08-26

    A novel concept for the preparation of multiphase composite ceramics based on demixing of a single ceramic precursor has been developed and used for the synthesis of a dual-phase H2 -permeable ceramic membrane. The precursor BaCe0.5 Fe0.5 O3-δ decomposes on calcination at 1370 °C for 10 h into two thermodynamically stable oxides with perovskite structures: the cerium-rich oxide BaCe0.85 Fe0.15 O3-δ (BCF8515) and the iron-rich oxide BaCe0.15 Fe0.85 O3-δ (BCF1585), 50 mol % each. In the resulting dual-phase material, the orthorhombic perovskite BCF8515 acts as the main proton conductor and the cubic perovskite BCF1585 as the main electron conductor. The dual-phase membrane shows an extremely high H2 permeation flux of 0.76 mL min(-1)  cm(-2) at 950 °C with 1.0 mm thickness. This auto-demixing concept should be applicable to the synthesis of other ionic-electronic conducting ceramics. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Comparison of potentially pathogenic free-living amoeba hosts by Legionella spp. in substrate-associated biofilms and floating biofilms from spring environments.

    PubMed

    Hsu, Bing-Mu; Huang, Chin-Chun; Chen, Jung-Sheng; Chen, Nai-Hsiung; Huang, Jen-Te

    2011-10-15

    This study compares five genera of free-living amoebae (FLA) hosts by Legionella spp. in the fixed and floating biofilm samples from spring environments. Detection rate of Legionella spp. was 26.9% for the floating biofilms and 3.1% for the fixed biofilms. Acanthamoeba spp., Hartmanella vermiformis, and Naegleria spp. were more frequently detected in floating biofilm than in fixed biofilm samples. The percentage of pathogenic Acanthamoeba spp. among all the genus Acanthamoeba detected positive samples was 19.6%. The potential pathogenic Naegleria spp. (for example, Naegleria australiensis, Naegleria philippinensis, and Naegleria italica) was 54.2% to all the Naegleria detected positive samples. In the study, 12 serotypes of possible pneumonia causing Legionella spp. were detected, and their percentage in all the Legionella containing samples was 42.4%. The FLA parasitized by Legionella included unnamed Acanthamoeba genotype, Acanthamoeba griffini, Acanthamoeba jacobsi, H. vermiformis, and N. australiensis. Significant differences were also observed between the presence/absence of H. vermiformis and Legionella parasitism in FLA. Comparisons between the culture-confirmed method and the PCR-based detection method for detecting FLA and Legionella in biofilms showed great variation. Therefore, using these analysis methods together to detect FLA and Legionella is recommended. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Diffusion of antimicrobials in multispecies biofilms evaluated in a new biofilm model.

    PubMed

    van der Waal, S V; de Almeida, J; Krom, B P; de Soet, J J; Crielaard, W

    2017-04-01

    To describe the application of a newly-developed in vitro model in which the diffusion of antimicrobials in oral biofilms can be studied. In a flow chamber consisting of three parallel feeding channels connected with each other by eight perpendicular side channels, multispecies biofilms were grown from saliva of a single donor for 48 h. The dimensions of the side channels were 100 μm × 100 μm × 5130 μm (H × W × L). When one or more side channels were filled with biofilm, the biofilms were stained with fluorescent stains. Then, one side-channel biofilm was selected and treated with phosphate buffered saline, 2% sodium hypochlorite (NaOCl), 17% ethylenediaminetetra-acetic acid (EDTA) or modified salt solution (MSS). Diffusion of the irrigants was observed by acquiring fluorescence images at 10× objective every 15 s for 30 min. It was possible to culture biofilms in the narrow (100 μm) channels. The biofilms varied in phenotype. In this model, no diffusion of NaOCl into the biofilms was seen after its application. Seventeen-percentage EDTA only diffused into the biofilm up to 200 μm in 30 min. MSS did diffuse in the biofilm over a distance of 450 μm within 2 min after a single application. This new model enables the investigation of the diffusion of antimicrobials in biofilms. Other applications to improve our understanding of the characteristics of biofilms are now possible. © 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  13. Biological treatment of mixtures of toluene and n-hexane vapours in a hollow fibre membrane bioreactor.

    PubMed

    Zhao, Kang; Xiu, Guangli; Xu, Lihang; Zhang, Danian; Zhang, Xiaofeng; Deshusses, Marc A

    2011-04-01

    Membrane bioreactors are gaining interest for the control of contaminated air streams. In this study, the removal of toluene and n-hexane vapours in a hollow fibre membrane bioreactor (HFMB) was investigated. The focus was on quantifying the possible interactions occurring during the simultaneous biotreatment of the two volatile pollutants. Two lab-scale units fitted with microporous polypropylene hollow fibre membranes were connected in series and inoculated with activated sludge. Contaminated air was passed through the lumen at gas residence times ranging from 2.3 to 9.4 s while a pollutant-degrading biofilm developed on the shell side of the fibres. When toluene was treated alone, very high elimination capacities (up to 750 g m(-3) h(-1) based on lumen volume, or 1.25 g m(-2) h(-1) when normalized by the hollow fibre membrane area) were reached. When toluene and hexane were treated simultaneously, toluene biodegradation was partially inhibited by n-hexane, resulting in lower toluene removal rates. On the other hand, hexane removal was only marginally affected by the presence of toluene and was degraded at very high rates (upwards of 440 g m(-3) h(-1) or 0.73 g m(-2) h(-1) without breakthrough). Overall, this study demonstrates that mixtures of toluene and n-hexane vapours can be effectively removed in hollow fibre membrane bioreactors and that complex biological interactions may affect one or more of the pollutants undergoing treatment in gas-phase membrane bioreactors.

  14. Archaeal Diversity in Biofilm Technologies Applied to Treat Urban and Industrial Wastewater: Recent Advances and Future Prospects

    PubMed Central

    Calderón, Kadiya; González-Martínez, Alejandro; Gómez-Silván, Cinta; Osorio, Francisco; Rodelas, Belén; González-López, Jesús

    2013-01-01

    Biological wastewater treatment (WWT) frequently relies on biofilms for the removal of anthropogenic contaminants. The use of inert carrier materials to support biofilm development is often required, although under certain operating conditions microorganisms yield structures called granules, dense aggregates of self-immobilized cells with the characteristics of biofilms maintained in suspension. Molecular techniques have been successfully applied in recent years to identify the prokaryotic communities inhabiting biofilms in WWT plants. Although methanogenic Archaea are widely acknowledged as key players for the degradation of organic matter in anaerobic bioreactors, other biotechnological functions fulfilled by Archaea are less explored, and research on their significance and potential for WWT is largely needed. In addition, the occurrence of biofilms in WWT plants can sometimes be a source of operational problems. This is the case for membrane bioreactors (MBR), an advanced technology that combines conventional biological treatment with membrane filtration, which is strongly limited by biofouling, defined as the undesirable accumulation of microbial biofilms and other materials on membrane surfaces. The prevalence and spatial distribution of archaeal communities in biofilm-based WWT as well as their role in biofouling are reviewed here, in order to illustrate the significance of this prokaryotic cellular lineage in engineered environments devoted to WWT. PMID:24022691

  15. Mitigation of Biofilm Development on Thin-Film Composite Membranes Functionalized with Zwitterionic Polymers and Silver Nanoparticles.

    PubMed

    Liu, Caihong; Faria, Andreia F; Ma, Jun; Elimelech, Menachem

    2017-01-03

    We demonstrate the functionalization of thin-film composite membranes with zwitterionic polymers and silver nanoparticles (AgNPs) for combating biofouling. Combining hydrophilic zwitterionic polymer brushes and biocidal AgNPs endows the membrane with dual functionality: antiadhesion and bacterial inactivation. An atom transfer radical polymerization (ATRP) reaction is used to graft zwitterionic poly(sulfobetaine methacrylate) (PSBMA) brushes to the membrane surface, while AgNPs are synthesized in situ through chemical reduction of silver. Two different membrane architectures (Ag-PSBMA and PSBMA-Ag TFC) are developed according to the sequence AgNPs, and PSBMA brushes are grafted on the membrane surface. A static adhesion assay shows that both modified membranes significantly reduced the adsorption of proteins, which served as a model organic foulant. However, improved antimicrobial activity is observed for PSBMA-Ag TFC (i.e., AgNPs on top of the polymer brush) in comparison to the Ag-PSBMA TFC membrane (i.e., polymer brush on top of AgNPs), indicating that architecture of the antifouling layer is an important factor in the design of zwitterion-silver membranes. Confocal laser scanning microscopy (CLSM) imaging indicated that PSBMA-Ag TFC membranes effectively inhibit biofilm formation under dynamic cross-flow membrane biofouling tests. Finally, we demonstrate the regeneration of AgNPs on the membrane after depletion of silver from the surface of the PSBMA-Ag TFC membrane.

  16. Inhibition and Dispersal of Pseudomonas aeruginosa Biofilms by Combination Treatment with Escapin Intermediate Products and Hydrogen Peroxide.

    PubMed

    Santiago, Ariel J; Ahmed, Marwa N A; Wang, Shu-Lin; Damera, Krishna; Wang, Binghe; Tai, Phang C; Gilbert, Eric S; Derby, Charles D

    2016-09-01

    Escapin is an l-amino acid oxidase that acts on lysine to produce hydrogen peroxide (H2O2), ammonia, and equilibrium mixtures of several organic acids collectively called escapin intermediate products (EIP). Previous work showed that the combination of synthetic EIP and H2O2 functions synergistically as an antimicrobial toward diverse planktonic bacteria. We initiated the present study to investigate how the combination of EIP and H2O2 affected bacterial biofilms, using Pseudomonas aeruginosa as a model. Specifically, we examined concentrations of EIP and H2O2 that inhibited biofilm formation or fostered disruption of established biofilms. High-throughput assays of biofilm formation using microtiter plates and crystal violet staining showed a significant effect from pairing EIP and H2O2, resulting in inhibition of biofilm formation relative to biofilm formation in untreated controls or with EIP or H2O2 alone. Similarly, flow cell analysis and confocal laser scanning microscopy revealed that the EIP and H2O2 combination reduced the biomass of established biofilms relative to that of the controls. Area layer analysis of biofilms posttreatment indicated that disruption of biomass occurs down to the substratum. Only nanomolar to micromolar concentrations of EIP and H2O2 were required to impact biofilm formation or disruption, and these concentrations are significantly lower than those causing bactericidal effects on planktonic bacteria. Micromolar concentrations of EIP and H2O2 combined enhanced P. aeruginosa swimming motility compared to the effect of either EIP or H2O2 alone. Collectively, our results suggest that the combination of EIP and H2O2 may affect biofilms by interfering with bacterial attachment and destabilizing the biofilm matrix. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  17. Post-deposition annealing temperature dependence TiO{sub 2}-based EGFET pH sensor sensitivity

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zulkefle, M. A., E-mail: alhadizulkefle@gmail.com; Rahman, R. A., E-mail: rohanieza.abdrahman@gmail.com; Yusoff, K. A., E-mail: khairul.aimi.yusof@gmail.com

    EGFET pH sensor is one type of pH sensor that is used to measure and determine pH of a solution. The sensing membrane of EGFET pH sensor plays vital role in the overall performance of the sensor. This paper studies the effects of different annealing temperature of the TiO{sub 2} sensing membranes towards sensitivity of EGFET pH sensor. Sol-gel spin coating was chosen as TiO{sub 2} deposition techniques since it is cost-effective and produces thin film with uniform thickness. Deposited TiO{sub 2} thin films were then annealed at different annealing temperatures and then were connected to the gate of MOSFETmore » as a part of the EGFET pH sensor structure. The thin films now act as sensing membranes of the EGFET pH sensor and sensitivity of each sensing membrane towards pH was measured. From the results it was determined that sensing membrane annealed at 300 °C gave the highest sensitivity followed by sample annealed at 400 °C and 500 °C.« less

  18. Corrosive effects of fluoride on titanium under artificial biofilm.

    PubMed

    Fukushima, Azusa; Mayanagi, Gen; Sasaki, Keiichi; Takahashi, Nobuhiro

    2018-01-01

    This study aimed to investigate the effect of sodium fluoride (NaF) on titanium corrosion using a biofilm model, taking environmental pH into account. Streptococcus mutans cells were used as the artificial biofilm, and pH at the bacteria-titanium interface was monitored after the addition of 1% glucose with NaF (0, 225 or 900ppmF) at 37°C for 90min. In an immersion test, the titanium samples were immersed in the NaF solution (0, 225 or 900ppm F; pH 4.2 or 6.5) for 30 or 90min. Before and after pH monitoring or immersion test, the electrochemical properties of the titanium surface were measured using a potentiostat. The amount of titanium eluted into the biofilm or the immersion solution was measured using inductively coupled plasma mass spectrometry. The color difference (ΔE*ab) and gloss of the titanium surface were determined using a spectrophotometer. After incubation with biofilm, pH was maintained at around 6.5 in the presence of NaF. There was no significant change in titanium surface and elution, regardless of the concentration of NaF. After immersion in 900ppm NaF solution at pH 4.2, corrosive electrochemical change was induced on the surface, titanium elution and ΔE*ab were increased, and gloss was decreased. NaF induces titanium corrosion in acidic environment in vitro, while NaF does not induce titanium corrosion under the biofilm because fluoride inhibits bacterial acid production. Neutral pH fluoridated agents may still be used to protect the remaining teeth, even when titanium-based prostheses are worn. Copyright © 2017 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.

  19. Rapid Evolution of Culture-Impaired Bacteria During Adaptation to Biofilm Growth

    PubMed Central

    Penterman, Jon; Nguyen, Dao; Anderson, Erin; Staudinger, Benjamin J.; Greenberg, Everett P.; Lam, Joseph S.; Singh, Pradeep K.

    2014-01-01

    Summary Biofilm growth increases the fitness of bacteria in harsh conditions. However, bacteria from clinical and environmental biofilms can exhibit impaired growth in culture, even when the species involved are readily cultureable, and permissive conditions are used. Here we show that culture-impaired variants of Pseudomonas aeruginosa rapidly and abundantly evolve in laboratory biofilms. The culture-impaired phenotype is caused by mutations that alter the outer-membrane lipopolysaccharide structure. Within biofilms, the lipopolysaccharide mutations markedly increase bacterial fitness. However, outside the protected biofilm environment, the mutations sensitize the variants to killing by a self-produced antimicrobial agent. Thus, a biofilm-mediated adaptation produces a stark fitness trade off that compromises bacterial survival in culture. Trade offs like this could limit the ability of bacteria to transition between biofilm growth and the free-living state, and produce bacterial populations that escape detection by culture-based sampling. PMID:24412364

  20. Nanoparticle-based Therapies for Wound Biofilm Infection: Opportunities and Challenges

    PubMed Central

    Kim, Min-Ho

    2016-01-01

    Clinical data from human chronic wounds implicates biofilm formation with the onset of wound chronicity. Despite the development of novel antimicrobial agents, the cost and complexity of treating chronic wound infections associated with biofilms remain a serious challenge, which necessitates the development of new and alternative approaches for effective anti-biofilm treatment. Recent advancement in nanotechnology for developing a new class of nanoparticles that exhibit unique chemical and physical properties holds promise for the treatment of biofilm infections. Over the last decade, nanoparticle-based approaches against wound biofilm infection have been directed toward developing nanoparticles with intrinsic antimicrobial properties, utilizing nanoparticles for controlled antimicrobials delivery, and applying nanoparticles for antibacterial hyperthermia therapy. In addition, a strategy to functionalize nanoparticles towards enhanced penetration through the biofilm matrix has been receiving considerable interest recently by means of achieving an efficient targeting to the bacterial cells within biofilm matrix. This review summarizes and highlights the recent development of these nanoparticle-based approaches as potential therapeutics for controlling wound biofilm infection, along with current challenges that need to be overcome for their successful clinical translation. PMID:26955044

  1. Algae-based biofilm productivity utilizing dairy wastewater: effects of temperature and organic carbon concentration.

    PubMed

    Fica, Zachary T; Sims, Ronald C

    2016-01-01

    Biofilm-based microalgal growth was determined as functions of organic chemical loading and water temperature utilizing dairy wastewater from a full-scale dairy farm. The dairy industry is a significant source of wastewater worldwide that could provide an inexpensive and nutrient rich feedstock for the cultivation of algae biomass for use in downstream processing of animal feed and aquaculture applications. Algal biomass was cultivated using a Rotating Algal Biofilm Reactor (RABR) system. The RABR is a biofilm-based technology that has been designed and used to remediate municipal wastewater and was applied to treat dairy wastewater through nutrient uptake, and simultaneously provide biomass for the production of renewable bioproducts. Aerial algal biofilm growth rates in dairy wastewater at 7 and 27 °C temperatures were shown to be 4.55 ± 0.17 g/m 2 -day and 7.57 ± 1.12 g/m 2 -day ash free dry weight (AFDW), respectively. Analysis of Variance (ANOVA) calculations indicated that both an increase in temperature of the wastewater and an increase in the level of organic carbon, from 300 to 1200 mg L -1 , contributed significantly to an increase in the rate of biomass growth in the system. However, ANOVA results indicated that the interaction of temperature and organic carbon content was not significantly related to the biofilm-based growth rate. A microalgae-based biofilm reactor was successfully used to treat turbid dairy wastewater. Temperature and organic carbon concentration had a statistically significant effect on algae-based biofilm productivity and treatment of dairy wastewater. The relationships between temperature, TOC, and productivity developed in this study may be used in the design and assessment of wastewater remediation systems and biomass production systems utilizing algae-based biofilm reactors for treating dairy wastes.

  2. The diphtheria toxin transmembrane domain as a pH sensitive membrane anchor for human interleukin-2 and murine interleukin-3.

    PubMed

    Liger, D; Nizard, P; Gaillard, C; vanderSpek, J C; Murphy, J R; Pitard, B; Gillet, D

    1998-11-01

    We have constructed two fusion proteins T-hIL-2 and T-mIL-3 in which human interleukin-2 (hIL-2) or murine interleukin-3 (mIL-3) are fused to the C-terminus of the diphtheria toxin transmembrane domain (T domain). Two additional fusion proteins, T-(Gly4-Ser)2-hIL-2 and T-(Gly4-Ser)2-mIL-3, were derived by introduction of the (Gly4-Ser)2 spacer between the T domain and cytokine components. Recognition of the hIL-2 receptor or the mIL-3 receptor by the corresponding recombinant proteins was demonstrated by their capacity to stimulate cytokine-dependent cell lines. All proteins retained the capacity of the T domain to insert into phospholipid membranes at acidic pH. Finally, anchoring of both cytokines to the membrane of lipid vesicles or living cells was assessed by specific antibody recognition. Our results show that the T domain fused to the N-terminus of a given protein can function as a pH sensitive membrane anchor for that protein.

  3. Inactivation of biofilm bacteria.

    PubMed Central

    LeChevallier, M W; Cawthon, C D; Lee, R G

    1988-01-01

    The current project was developed to examine inactivation of biofilm bacteria and to characterize the interaction of biocides with pipe surfaces. Unattached bacteria were quite susceptible to the variety of disinfectants tested. Viable bacterial counts were reduced 99% by exposure to 0.08 mg of hypochlorous acid (pH 7.0) per liter (1 to 2 degrees C) for 1 min. For monochloramine, 94 mg/liter was required to kill 99% of the bacteria within 1 min. These results were consistent with those found by other investigators. Biofilm bacteria grown on the surfaces of granular activated carbon particles, metal coupons, or glass microscope slides were 150 to more than 3,000 times more resistant to hypochlorous acid (free chlorine, pH 7.0) than were unattached cells. In contrast, resistance of biofilm bacteria to monochloramine disinfection ranged from 2- to 100-fold more than that of unattached cells. The results suggested that, relative to inactivation of unattached bacteria, monochloramine was better able to penetrate and kill biofilm bacteria than free chlorine. For free chlorine, the data indicated that transport of the disinfectant into the biofilm was a major rate-limiting factor. Because of this phenomenon, increasing the level of free chlorine did not increase disinfection efficiency. Experiments where equal weights of disinfectants were used suggested that the greater penetrating power of monochloramine compensated for its limited disinfection activity. These studies showed that monochloramine was as effective as free chlorine for inactivation of biofilm bacteria. The research provides important insights into strategies for control of biofilm bacteria. Images PMID:2849380

  4. Simultaneous enrichment of denitrifying anaerobic methane-oxidizing microorganisms and anammox bacteria in a hollow-fiber membrane biofilm reactor.

    PubMed

    Ding, Zhao-Wei; Lu, Yong-Ze; Fu, Liang; Ding, Jing; Zeng, Raymond J

    2017-01-01

    In this study, the coculture system of denitrifying anaerobic methane oxidation (DAMO) microbes and anaerobic ammonium oxidation (anammox) bacteria was successfully enriched in a hollow-fiber membrane biofilm reactor (HfMBR) using freshwater sediment as the inoculum. The maximal removal rates of nitrate and ammonium were 78 mg N/L/day (131 mg N/m 2 /day) and 26 mg N/L/day (43 mg N/m 2 /day), respectively. Due to the high rate of methane mass transfer in HfMBR, the activity of DAMO archaea continued to increase during the enrichment period, indicating that HfMBR could be a powerful tool to enrich DAMO microorganisms. Effects of partial methane pressure, temperature, and pH on the cocultures were obvious. However, the microbial activity in HfMBR could be recovered quickly after the shock change of environmental factors. Furthermore, the result also found that DAMO bacteria likely had a stronger competitive advantage than anammox bacteria under the operating conditions in this study. High-throughput sequencing 16S rRNA genes illustrated that the dominant microbes were NC10, Euryarchaeota, Proteobacteria, Planctomycetes, and Chlorobi with relative abundance of 38.8, 26.2, 13.78, 6.2, and 3.6 %, respectively.

  5. Nonuniform spatial patterns of respiratory activity within biofilms during disinfection.

    PubMed Central

    Huang, C T; Yu, F P; McFeters, G A; Stewart, P S

    1995-01-01

    Fluorescent stains in conjunction with cryoembedding and image analysis were applied to demonstrate spatial gradients in respiratory activity within bacterial biofilms during disinfection with monochloramine. Biofilms of Klebsiella pneumoniae and Pseudomonas aeruginosa grown together on stainless steel surfaces in continuous-flow annular reactors were treated with 2 mg of monochloramine per liter (influent concentration) for 2 h. Relatively little biofilm removal occurred as evidenced by total cell direct counts. Plate counts (of both species summed) indicated an average 1.3-log decrease after exposure to 2 mg of monochloramine per liter. The fluorogenic redox indicator 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and the DNA stain 4',6-diamidino-2-phenylindole (DAPI) were used to differentiate respiring and nonrespiring cells in biofilms. Epifluorescence micrographs of frozen biofilm cross sections clearly revealed gradients of respiratory activity within biofilms in response to monochloramine treatment. These gradients in specific respiratory activity were quantified by calculating the ratio of CTC and DAPI intensities measured by image analysis. Cells near the biofilm-bulk fluid interface lost respiratory activity first. After 2 h of biocide treatment, greater respiratory activity persisted deep in the biofilm than near the biofilm-bulk fluid interface. PMID:7793945

  6. DNase I and proteinase K impair Listeria monocytogenes biofilm formation and induce dispersal of pre-existing biofilms.

    PubMed

    Nguyen, Uyen T; Burrows, Lori L

    2014-09-18

    Current sanitation methods in the food industry are not always sufficient for prevention or dispersal of Listeria monocytogenes biofilms. Here, we determined if prevention of adherence or dispersal of existing biofilms could occur if biofilm matrix components were disrupted enzymatically. Addition of DNase during biofilm formation reduced attachment (<50% of control) to polystyrene. Treatment of established 72h biofilms with 100μg/ml of DNase for 24h induced incomplete biofilm dispersal, with <25% biofilm remaining compared to control. In contrast, addition of proteinase K completely inhibited biofilm formation, and 72h biofilms-including those grown under stimulatory conditions-were completely dispersed with 100μg/ml proteinase K. Generally-regarded-as-safe proteases bromelain and papain were less effective dispersants than proteinase K. In a time course assay, complete dispersal of L. monocytogenes biofilms from both polystyrene and type 304H food-grade stainless steel occurred within 5min at proteinase K concentrations above 25μg/ml. These data confirm that both DNA and proteins are required for L. monocytogenes biofilm development and maintenance, and that these components of the biofilm matrix can be targeted for effective prevention and removal of biofilms. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Confocal laser scanning microscopy analysis of S. epidermidis biofilms exposed to farnesol, vancomycin and rifampicin.

    PubMed

    Cerca, Nuno; Gomes, Fernanda; Pereira, Sofia; Teixeira, Pilar; Oliveira, Rosário

    2012-05-16

    Staphylococcus epidermidis is the major bacterial species found in biofilm-related infections on indwelling medical devices. Microbial biofilms are communities of bacteria adhered to a surface and surrounded by an extracellular polymeric matrix. Biofilms have been associated with increased antibiotic tolerance to the immune system. This increased resistance to conventional antibiotic therapy has lead to the search for new antimicrobial therapeutical agents. Farnesol, a quorum-sensing molecule in Candida albicans, has been described as impairing growth of several different microorganisms and we have previously shown its potential as an adjuvant in antimicrobial therapy against S. epidermidis. However, its mechanism of action in S. epidermidis is not fully known. In this work we better elucidate the role of farnesol against S: epidermidis biofilms using confocal laser scanning microscopy (CLSM). 24 h biofilms were exposed to farnesol, vancomycin or rifampicin and were analysed by CLSM, after stained with a Live/Dead stain, a known indicator of cell viability, related with cell membrane integrity. Biofilms were also disrupted by sonication and viable and cultivable cells were quantified by colony forming units (CFU) plating. Farnesol showed a similar effect as vancomycin, both causing little reduction of cell viability but at the same time inducing significant changes in the biofilm structure. On the other hand, rifampicin showed a distinct action in S. epidermidis biofilms, by killing a significant proportion of biofilm bacteria. While farnesol is not very efficient at killing biofilm bacteria, it damages cell membrane, as determined by the live/dead staining, in a similar way as vancomycin. Furthermore, farnesol might induce biofilm detachment, as determined by the reduced biofilm biomass, which can partially explain the previous findings regarding its role as a possible chemotherapy adjuvant.

  8. Microbial Adhesion and Biofilm Formation on Microfiltration Membranes: A Detailed Characterization Using Model Organisms with Increasing Complexity

    PubMed Central

    Vanysacker, L.; Denis, C.; Declerck, P.; Piasecka, A.; Vankelecom, I. F. J.

    2013-01-01

    Since many years, membrane biofouling has been described as the Achilles heel of membrane fouling. In the present study, an ecological assay was performed using model systems with increasing complexity: a monospecies assay using Pseudomonas aeruginosa or Escherichia coli separately, a duospecies assay using both microorganisms, and a multispecies assay using activated sludge with or without spiked P. aeruginosa. The microbial adhesion and biofilm formation were evaluated in terms of bacterial cell densities, species richness, and bacterial community composition on polyvinyldifluoride, polyethylene, and polysulfone membranes. The data show that biofouling formation was strongly influenced by the kind of microorganism, the interactions between the organisms, and the changes in environmental conditions whereas the membrane effect was less important. The findings obtained in this study suggest that more knowledge in species composition and microbial interactions is needed in order to understand the complex biofouling process. This is the first report describing the microbial interactions with a membrane during the biofouling development. PMID:23986906

  9. Microtubule-dependent relocation of branchial V-H+-ATPase to the basolateral membrane in the Pacific spiny dogfish (Squalus acanthias): a role in base secretion.

    PubMed

    Tresguerres, Martin; Parks, Scott K; Katoh, Fumi; Goss, Greg G

    2006-02-01

    We have previously shown that continuous intravenous infusion of NaHCO3 for 24 h ( approximately 1000 micromol kg(-1) h(-1)) results in the relocation of V-H+-ATPase from the cytoplasm to the basolateral membrane in the gills of the Pacific dogfish. To further investigate this putative base-secretive process we performed similar experiments with the addition of colchicine, an inhibitor of cytoskeleton-dependent cellular trafficking processes. Blood pH and plasma total CO2 were significantly higher in the colchicines-treated, HCO3- -infused fish compared with fish infused with HCO3- alone. The effect of colchicine was highest after 24 h of infusion (8.33+/-0.06 vs 8.02+/-0.03 pH units, 15.72+/-3.29 vs 6.74+/-1.34 mmol CO2 l(-1), N=5). Immunohistochemistry and western blotting confirmed that colchicine blocked the transit of V-H+-ATPase to the basolateral membrane. Furthermore, western blotting analyses from whole gill and cell membrane samples suggest that the short-term (6 h) response to alkaline stress consists of relocation of V-H+-ATPases already present in the cell to the basolateral membrane, while in the longer term (24 h) there is both relocation of preexistent enzyme and upregulation in the synthesis of new units. Our results strongly suggest that cellular relocation of V-H+-ATPase is necessary for enhanced HCO3- secretion across the gills of the Pacific dogfish.

  10. Use of Potential Probiotic Lactic Acid Bacteria (LAB) Biofilms for the Control of Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7 Biofilms Formation

    PubMed Central

    Gómez, Natacha C.; Ramiro, Juan M. P.; Quecan, Beatriz X. V.; de Melo Franco, Bernadette D. G.

    2016-01-01

    Use of probiotic biofilms can be an alternative approach for reducing the formation of pathogenic biofilms in food industries. The aims of this study were (i) to evaluate the probiotic properties of bacteriocinogenic (Lactococcus lactis VB69, L. lactis VB94, Lactobacillus sakei MBSa1, and Lactobacillus curvatus MBSa3) and non-bacteriocinogenic (L. lactis 368, Lactobacillus helveticus 354, Lactobacillus casei 40, and Weissela viridescens 113) lactic acid bacteria (LAB) isolated from Brazilian’s foods and (ii) to develop protective biofilms with these strains and test them for exclusion of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Typhimurium. LAB were tested for survival in acid and bile salt conditions, surface properties, biosurfactant production, β-galactosidase and gelatinase activity, antibiotic resistance and presence of virulence genes. Most strains survived exposure to pH 2 and 4% bile salts. The highest percentages of auto-aggregation were obtained after 24 h of incubation. Sixty-seven percentage auto-aggregation value was observed in W. viridescens 113 and Lactobacillus curvatus MBSa3 exhibited the highest co-aggregation (69% with Listeria monocytogenes and 74.6% with E. coli O157:H7), while the lowest co-aggregation was exhibited by W. viridescens 113 (53.4% with Listeria monocytogenes and 38% with E. coli O157:H7). Tests for hemolytic activity, bacterial cell adherence with xylene, and drop collapse confirmed the biosurfactant-producing ability of most strains. Only one strain (L. lactis 368) produced β-galactosidase. All strains were negative for virulence genes cob, ccf, cylLL, cylLs, cyllM, cylB, cylA and efaAfs and gelatinase production. The antibiotic susceptibility tests indicated that the MIC for ciprofloxacin, clindamycin, gentamicin, kanamycin, and streptomycin did not exceed the epidemiological cut-off suggested by the European Food Safety Authority. Some strains were resistant to one or more antibiotics and

  11. Biofilm formation by Shiga toxin-producing Escherichia coli O157:H7 and Non-O157 strains and their tolerance to sanitizers commonly used in the food processing environment.

    PubMed

    Wang, Rong; Bono, James L; Kalchayanand, Norasak; Shackelford, Steven; Harhay, Dayna M

    2012-08-01

    Shiga toxin-producing Escherichia coli (STEC) strains are important foodborne pathogens. Among these, E. coli O157:H7 is the most frequently isolated STEC serotype responsible for foodborne diseases. However, the non-O157 serotypes have been associated with serious outbreaks and sporadic diseases as well. It has been shown that various STEC serotypes are capable of forming biofilms on different food or food contact surfaces that, when detached, may lead to cross-contamination. Bacterial cells at biofilm stage also are more tolerant to sanitizers compared with their planktonic counterparts, which makes STEC biofilms a serious food safety concern. In the present study, we evaluated the potency of biofilm formation by a variety of STEC strains from serotypes O157:H7, O26:H11, and O111:H8; we also compared biofilm tolerance with two types of common sanitizers, a quaternary ammonium chloride-based sanitizer and chlorine. Our results demonstrated that biofilm formation by various STEC serotypes on a polystyrene surface was highly strain-dependent, whereas the two non-O157 serotypes showed a higher potency of pellicle formation at air-liquid interfaces on a glass surface compared with serotype O157:H7. Significant reductions of viable biofilm cells were achieved with sanitizer treatments. STEC biofilm tolerance to sanitization was strain-dependent regardless of the serotypes. Curli expression appeared to play a critical role in STEC biofilm formation and tolerance to sanitizers. Our data indicated that multiple factors, including bacterial serotype and strain, surface materials, and other environmental conditions, could significantly affect STEC biofilm formation. The high potential for biofilm formation by various STEC serotypes, especially the strong potency of pellicle formation by the curli-positive non-O157 strains with high sanitization tolerance, might contribute to bacterial colonization on food contact surfaces, which may result in downstream product

  12. The Major Outer Membrane Protein MopB Is Required for Twitching Movement and Affects Biofilm Formation and Virulence in Two Xylella fastidiosa strains.

    PubMed

    Chen, Hongyu; Kandel, Prem P; Cruz, Luisa F; Cobine, Paul A; De La Fuente, Leonardo

    2017-11-01

    MopB is a major outer membrane protein (OMP) in Xylella fastidiosa, a bacterial plant pathogen that causes losses on many economically important crops. Based on in silico analysis, the uncharacterized MopB protein of X. fastidiosa contains a β-barrel structure with an OmpA-like domain and a predicted calcium-binding motif. Here, MopB function was studied by mutational analysis taking advantage of the natural competence of X. fastidiosa. Mutants of mopB were constructed in two different X. fastidiosa strains, the type strain Temecula and the more virulent WM1-1. Deletion of the mopB gene impaired cell-to-cell aggregation, surface attachment, and biofilm formation in both strains. Interestingly, mopB deletion completely abolished twitching motility. Electron microscopy of the bacterial cell surface revealed that mopB deletion eliminated type IV and type I pili formation, potentially caused by destabilization of the outer membrane. Both mopB mutants showed reduced virulence using tobacco (Nicotiana tabacum) as a host under greenhouse conditions. These results suggest that MopB has pleiotropic functions in biofilm formation and twitching motility and is important for virulence of X. fastidiosa.

  13. [Confocal laser scanning electron microscopy for assessment of vaginal Lactobacillus crispatus biofilm].

    PubMed

    Wu, Li-jie; Wang, Ben; Liao, Qin-ping; Zhang, Rui

    2015-12-18

    To investigate the female vaginal Lactobacillus crispatus biofilm by using confocal laser scanning microscopy (CLSM),thus revealing the formation of biofilm. The cover slide biofilm culture approach in vitro was employed for induction of the vaginal Lactobacillus crispatus biofilm formation. Following the culture for 2, 4, 8, 12, 16, 20, 24, 48, 72, 96 and 120 hours, the cover slide was removed for subsequent staining with the fluoresce in isothiocyanate-conjugated concanavalin A(FITC-ConA) and propidium (PI).This was followed by determination of the formation and characteristics of the vaginal Lactobacillus crispatus biofilm by using CLSM. The CLSM images of biofilm formation at different time points were captured, suggesting that the vaginal Lactobacillus crispatus adhesion occurred at h 4, which was in reversible attachment, then more and more Lactobacillus crispatus aggregated at h 8 to h 20, which was in irreversible attachment.Lactobacillus crispatus clustered at h 20, with early development of biofilm architecture.Then the biofilm with extracellular matrix around the bacteria was set up at h 24,with gradual matureation at h 24 to h 48.The biofilm dispersed at h 72. The biofilm density of cultivating for 20 hours was 42.7 × 10⁻³ ± 6.8 × 10⁻³ ,and for 24 hours increased to 102.5 × 10⁻³ ± 23.1 × 10⁻³, suggesting a significant difference, P<0.05. This meant that mature biofilm was formed at h 24. The vaginal Lactobacillus crispatus is able to form typical biofilm with distinct developmental phases and architecture characteristics.Mature biofilm is formed at h 24 to h 48, then the biofilm begins to disperse.

  14. A novel osmosis membrane bioreactor-membrane distillation hybrid system for wastewater treatment and reuse.

    PubMed

    Nguyen, Nguyen Cong; Nguyen, Hau Thi; Chen, Shiao-Shing; Ngo, Huu Hao; Guo, Wenshan; Chan, Wen Hao; Ray, Saikat Sinha; Li, Chi-Wang; Hsu, Hung-Te

    2016-06-01

    A novel approach was designed to simultaneously enhance nutrient removal and reduce membrane fouling for wastewater treatment using an attached growth biofilm (AGB) integrated with an osmosis membrane bioreactor (OsMBR) system for the first time. In this study, a highly charged organic compound (HEDTA(3-)) was employed as a novel draw solution in the AGB-OsMBR system to obtain a low reverse salt flux, maintain a healthy environment for the microorganisms. The AGB-OsMBR system achieved a stable water flux of 3.62L/m(2)h, high nutrient removal of 99% and less fouling during a 60-day operation. Furthermore, the high salinity of diluted draw solution could be effectively recovered by membrane distillation (MD) process with salt rejection of 99.7%. The diluted draw solution was re-concentrated to its initial status (56.1mS/cm) at recovery of 9.8% after 6h. The work demonstrated that novel multi-barrier systems could produce high quality potable water from impaired streams. Copyright © 2016. Published by Elsevier Ltd.

  15. Membrane fouling in a submerged membrane bioreactor: effect of pH and its implications.

    PubMed

    Zhang, Ye; Zhang, Meijia; Wang, Fangyuan; Hong, Huachang; Wang, Aijun; Wang, Juan; Weng, Xuexiang; Lin, Hongjun

    2014-01-01

    The effect of pH on membrane fouling in a submerged membrane bioreactor (MBR) was investigated in this study. It was found that, pH increase slightly increased the resistance of virgin membrane and fouled membrane. Pore clogging resistance was quite low, which was not apparently affected by the pH variation. Lower pH resulted in higher adherence of sludge flocs on membrane surface. Thermodynamic analysis showed that a repulsive energy barrier existed in the process of the foulants approaching to membrane surface. This energy barrier would decrease with pH decreased, suggesting the existence of a critical pH below which the repulsive energy barrier would disappear, which would facilitate attachment of the foulants. The resistance of the formed cake layer would significantly increase with the feed pH. This result could be explained by the osmotic pressure mechanism. The obtained findings also provided important implications for membrane fouling mitigation in MBRs. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Effectiveness of Sanitizer D7(TM) against Escherichia coli O157:H7 and Salmonella biofilms

    USDA-ARS?s Scientific Manuscript database

    Introduction: Biofilm formation by E. coli O157:H7 and Salmonella enterica at meat processing plants poses a serious risk of meat product contamination. Available studies have shown that many common sanitizers were unable to completely eradicate biofilms by these foodborne pathogens due to the 3-dim...

  17. Hydration of sulfonated polyimide membranes. I. Na + and NH +(C 2H 5) 3 homopolymers

    NASA Astrophysics Data System (ADS)

    Jamróz, Dorota; Maréchal, Yves

    2004-05-01

    Hydration of Na + and HN +(C 2H 5) 3 sulfonated polyimide membranes is probed by infrared spectrometry using a recently described method. These membranes consist of identical sulfonated repeat unit (homopolymers) and form a good starting point to study more elaborated membranes, composed of these units plus similar ones with no sulfonic groups (block copolymers). These latter membranes may be used in fuel cells and will be described in a forthcoming article. We first identify the bands of hydrophilic groups, which will be anchor points for hydration. We then define three 'elementary hydration spectra' onto which all hydration spectra can be decomposed. Their analysis allows us to measure the water uptake of these membranes as a function of the hygrometry of the ambient atmosphere and to determine the structures of the dried membranes and their hydration mechanisms in terms of chemical reactions.

  18. Ionic liquid-based materials: a platform to design engineered CO2 separation membranes.

    PubMed

    Tomé, Liliana C; Marrucho, Isabel M

    2016-05-21

    During the past decade, significant advances in ionic liquid-based materials for the development of CO2 separation membranes have been accomplished. This review presents a perspective on different strategies that use ionic liquid-based materials as a unique tuneable platform to design task-specific advanced materials for CO2 separation membranes. Based on compilation and analysis of the data hitherto reported, we provide a judicious assessment of the CO2 separation efficiency of different membranes, and highlight breakthroughs and key challenges in this field. In particular, configurations such as supported ionic liquid membranes, polymer/ionic liquid composite membranes, gelled ionic liquid membranes and poly(ionic liquid)-based membranes are detailed, discussed and evaluated in terms of their efficiency, which is attributed to their chemical and structural features. Finally, an integrated perspective on technology, economy and sustainability is provided.

  19. Recolonization of laser-ablated bacterial biofilm.

    PubMed

    Nandakumar, Kanavillil; Obika, Hideki; Utsumi, Akihiro; Toshihiko, Ooie; Yano, Tetsuo

    2004-01-20

    The recolonization of laser-ablated bacterial monoculture biofilm was studied in the laboratory by using a flow-cytometer system. The marine biofilm-forming bacterium Pseudoalteromonas carrageenovora was used to develop biofilms on titanium coupons. Upon exposure to a low-power pulsed irradiation from an Nd:YAG laser, the coupons with biofilm were significantly reduced both in terms of total viable count (TVC) and area cover. The energy density used for a pulse of 5 ns was 0.1 J/cm(2) and the durations of irradiation exposure were 5 and 10 min. When placed in a flow of dilute ZoBell marine broth medium (10%) the laser-destructed bacterial film in a flow-cytometer showed significant recovery over a period of time. The flow of medium was regulated at 3.2 ml/min. The increase in area cover and TVC, however, was significantly less than that observed for nonirradiated control (t-test, P< 0.05). The coupons were observed for biofilm area cover and TVC at different intervals (3, 6, and 9 h) after irradiation. While the biofilm in the control coupon at the end of 9 h of exposure showed 95.6 +/- 4.1% cover, the 5- and 10-min irradiated samples after 9 h showed 60.3 +/- 6.5 and 37.4 +/- 12.1% area cover, respectively. The reduced rate of recolonization compared to control was thought be due to the lethal and sublethal impacts of laser irradiation on bacteria. This observation thus provided data on the online recolonization speed of biofilm, which is important when considering pulsed laser irradiation as an ablating technique of biofilm formation and removal in natural systems. Copyright 2003 Wiley Periodicals, Inc.

  20. Influence of matrix and filler fraction on biofilm formation on the surface of experimental resin-based composites.

    PubMed

    Ionescu, Andrei; Brambilla, Eugenio; Wastl, Daniel S; Giessibl, Franz J; Cazzaniga, Gloria; Schneider-Feyrer, Sibylle; Hahnel, Sebastian

    2015-01-01

    The aim of this study was to investigate the impact of resin matrix chemistry and filler fraction on biofilm formation on the surface of experimental resin-based composites (RBCs). Specimens were prepared from eight experimental RBC formulations differing in resin matrix blend (BisGMA/TEGDMA in a 7:3 wt% ratio or UDMA/aliphatic dimethacrylate in a 1:1 wt% ratio) and filler fraction (no fillers; 65 wt% dental glass with an average diameter of 7 or 0.7 µm or 65 wt% SiO2 with an average diameter of 20 nm). Surface roughness, surface free energy, and chemical surface composition were determined; surface topography was visualized using atomic force microscopy. Biofilm formation was simulated under continuous flow conditions for a 48 h period using a monospecies Streptococcus mutans and a multispecies biofilm model. In the monospecies biofilm model, the impact of the filler fraction overruled the influence of the resin matrix, indicating lowest biofilm formation on RBCs with nano-scaled filler particles and those manufactured from the neat resin blends. The multispecies model suggested a more pronounced effect of the resin matrix blend, as significantly higher biofilm formation was identified on RBCs with a UDMA/dimethacrylate matrix blend than on those including a BisGMA/TEGDMA matrix blend but analogous filler fractions. Although significant differences in surface properties between the various materials were identified, correlations between the surface properties and biofilm formation were poor, which highlights the relevance of surface topography and chemistry. These results may help to tailor novel RBC formulations which feature reduced biofilm formation on their surface.

  1. Induction of a proton gradient across a gold-supported biomimetic membrane by electroenzymatic H2 oxidation.

    PubMed

    Gutiérrez-Sanz, Óscar; Tapia, Cristina; Marques, Marta C; Zacarias, Sonia; Vélez, Marisela; Pereira, Inês A C; De Lacey, Antonio L

    2015-02-23

    Energy-transduction mechanisms in living organisms, such as photosynthesis and respiration, store light and chemical energy in the form of an electrochemical gradient created across a lipid bilayer. Herein we show that the proton concentration at an electrode/phospholipid-bilayer interface can be controlled and monitored electrochemically by immobilizing a membrane-bound hydrogenase. Thus, the energy derived from the electroenzymatic oxidation of H2 can be used to generate a proton gradient across the supported biomimetic membrane. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Differential effects of antifungal agents on expression of genes related to formation of Candida albicans biofilms.

    PubMed

    Chatzimoschou, Athanasios; Simitsopoulou, Maria; Antachopoulos, Charalampos; Walsh, Thomas J; Roilides, Emmanuel

    2016-01-01

    The purpose of this study was to analyse specific molecular mechanisms involved in the intrinsic resistance of C. albicans biofilms to antifungals. We investigated the transcriptional profile of three genes (BGL2, SUN41, ECE1) involved in Candida cell wall formation in response to voriconazole or anidulafungin after the production of intermediate and mature biofilms. C. albicans M61, a well-documented biofilm producer strain, was used for the development of intermediate (12 h and 18 h) and completely mature biofilms (48 h). After exposure of cells from each biofilm growth mode to voriconazole (128 and 512 mg l(-1)) or anidulafungin (0.25 and 1 mg l(-1)) for 12-24 h, total RNA samples extracted from biofilm cells were analysed by RT-PCR. The voriconazole and anidulafungin biofilm MIC was 512 and 0.5 mg l(-1) respectively. Anidulafungin caused significant up-regulation of SUN41 (3.7-9.3-fold) and BGL2 (2.2-2.8 fold) in intermediately mature biofilms; whereas, voriconazole increased gene expression in completely mature biofilms (SUN41 2.3-fold, BGL2 2.1-fold). Gene expression was primarily down-regulated by voriconazole in intermediately, but not completely mature biofilms. Both antifungals caused down-regulation of ECE1 in intermediately mature biofilms. © 2015 Blackwell Verlag GmbH.

  3. Polyethylene Based Materials for Biofilm Carriers Used in Wastewater Treatment

    NASA Astrophysics Data System (ADS)

    Moga, I. C.; Iordache, O. I.; Petrescu, G.; Pricop, F.; Dumitrescu, I.

    2018-06-01

    The moving bed biofilm technology is based on biofilm carriers on which consortia of microorganisms attach, develop and grow. Around the world are known many biofilm carrier variants made of varied materials. The most common materials are based on polyethylene since this material has a close to water density. The authors propose a novel biofilm carrier to be used in tertiary treatment for tannery and paper-mill wastewaters. The biological treatment is based on fungal activity. The selected fungal strains will be grown on innovative polyethylene carriers containing cellulose. The carrier will be designed to be exploited in a moving bed bioreactor and to favour fungal growth in the presence of competing bacteria.

  4. Combination of synchrotron radiation-based Fourier transforms infrared microspectroscopy and confocal laser scanning microscopy to understand spatial heterogeneity in aquatic multispecies biofilms.

    PubMed

    Reuben, Sheela; Banas, Krzysztof; Banas, Agnieszka; Swarup, Sanjay

    2014-11-01

    Understanding the spatial heterogeneity within environmental biofilms can provide an insight into compartmentalization of different functions in biofilm communities. We used a non-destructive and label-free method by combining Synchrotron Radiation-based Fourier Transform Infrared Microspectroscopy (SR-FTIR) with Confocal Laser Scanning Microscopy (CLSM) to distinguish the spatial chemical changes within multispecies biofilms grown from natural storm waters in flow cells. Among the different surfaces tested for biofilm growth and optimal imaging, mylar membranes were most suited and it enabled successful spatial infrared imaging of natural biofilms for obtaining reliable and interpretable FTIR spectra. Time series analysis of biofilm growth showed that influx of water during biofilm growth, results in significant changes in biofilm formation. Early biofilms showed active nutrient acquisition and desiccation tolerance mechanisms corresponding with accumulation of secreted proteins. Statistical approach used for the evaluation of chemical spectra allowed for clustering and classification of various regions of the biofilm. Microheterogeneity was observed in the polymeric components of the biofilm matrix, including cellulose, glycocalyx and dextran-like molecules. Fructan and glycan-rich regions were distinguishable and glycocalyx was abundant in the strongly adhering peripheral regions of biofilms. Inner core showed coexistence of oxygen dimers and ferrihydrite that will likely support growth of Fe (II)-oxidising bacteria. The combined SR-FTIR microspectroscopy and CSLM approach for complex natural biofilms described here will be useful both in understanding heterogeneity of matrix components and in correlating functions of juxtaposed microbial species in complex natural biofilms with physicochemical microenvironment to which they are exposed. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation.

    PubMed

    Okumura, Masaki; Inoue, Shin-Ichiro; Kuwata, Keiko; Kinoshita, Toshinori

    2016-05-01

    Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. © 2016 American Society of Plant Biologists. All Rights Reserved.

  6. Biofilm formation in Haemophilus parasuis: relationship with antibiotic resistance, serotype and genetic typing.

    PubMed

    Zhang, Jianmin; Xu, Chenggang; Shen, Haiyan; Li, Jingyi; Guo, Lili; Cao, Guojie; Feng, Saixiang; Liao, Ming

    2014-10-01

    Biofilms are surface-associated microbial communities, which are encased in self-synthesized extracellular environment. Biofilm formation may trigger drug resistance and inflammation, resulting in persistent infections. Haemophilus parasuis is the etiological agent of a systemic disease, Glässer's disease, characterized by fibrinous polyserositis, arthritis and meningitis in pigs. The purpose of this study was to examine the correlation between biofilm and antibiotic resistance among the clinical isolates of H. parasuis. In the present study, we tested biofilm-forming ability of 110 H. parasuis isolates from various farms using polystyrene microtiter plate assays. Seventy-three isolates of H. parasuis (66.4%) showed biofilm formation and most of them performed weak biofilm-forming ability (38/73). All isolates were tested for antimicrobial susceptibility to 18 antimicrobial agents by the broth microdilution method. H. parasuis isolates showed very high resistance (>90%) to sulfanilamide, nalidixic acid, and trimethoprim. Resistance to eight antibiotics such as penicillin (41.1% vs 8.1%), ampicillin (31.5% vs 8.1%), amoxicillin (28.8% vs 5.4%), gentamicin (46.6% vs 24.3%), cefazolin (19.2% vs 2.7%), doxycycline (19.2% vs 8.1%), cefotaxime (11% vs 2.7%), and cefaclor (13.7% vs 5.4%) was comparatively higher among biofilm producers than non-biofilm producers. Pulsed-field gel electrophoresis (PFGE) analyses could distinguish various isolates. Our data indicated that H. parasuis field isolates were able to form biofilms in vitro. In addition, biofilm positive strains had positive correlation with resistance to β-lactams antibiotics. Thus, biofilm formation may play important roles during H. parasuis infections. Copyright © 2014. Published by Elsevier Ltd.

  7. Demetallization of Enterococcus faecalis biofilm: a preliminary study

    PubMed Central

    ESTRELA, Carlos; COSTA E SILVA, Rodrigo; URBAN, Roberta Cerasi; GONÇALVES, Pablo José; SILVA, Júlio A.; ESTRELA, Cyntia R.A.; PECORA, Jesus Djalma; PETERS, Ove A.

    2018-01-01

    Abstract Objectives To determine the concentration of calcium, iron, manganese and zinc ions after the application of chelator to Enterococcus faecalis biofilms. Material and Methods Fifty bovine maxillary central incisors were prepared and inoculated with E. faecalis for 60 days. The following were used as irrigation solutions: 17% EDTA (pH 3, 7 and 10), 2.5% sodium hypochlorite (NaOCl) combined with 17% EDTA (pH 3, 7 and 10), distilled water (pH 3, 7 and 10), and 2.5% NaOCl. Each solution was kept in the root canal for five minutes. Fifteen uncontaminated root canals were irrigated with 17% EDTA (pH 3, 7 and 10). Six teeth were used as bacterial control. The number of calcium, iron, manganese and zinc ions was determined using flame atomic absorption spectrometry. Mean ± standard deviation (SD) values were used for descriptive statistics. Results Calcium chelation using 17% EDTA at pH 7 was higher than at pH 3 and 10, regardless of whether bacterial biofilm was present. The highest concentration of iron occurred at pH 3 in the presence of bacterial biofilm. The highest concentration of manganese found was 2.5% NaOCl and 17% EDTA at pH 7 in the presence of bacterial biofilm. Zinc levels were not detectable. Conclusions The pH of chelating agents affected the removal of calcium, iron, and manganese ions. The concentration of iron ions in root canals with bacterial biofilm was higher after the use of 17% EDTA at pH 3 than after the use of the other solutions at all pH levels. PMID:29451651

  8. Integrated nitrogen removal biofilter system with ceramic membrane for advanced post-treatment of municipal wastewater.

    PubMed

    Son, Dong-Jin; Yun, Chan-Young; Kim, Woo-Yeol; Zhang, Xing-Ya; Kim, Dae-Gun; Chang, Duk; Sunwoo, Young; Hong, Ki-Ho

    2016-12-01

    The pre-denitrification biofilm process for nitrogen removal was combined with ceramic membrane with pore sizes of 0.05-0.1 µm as a system for advanced post-treatment of municipal wastewater. The system was operated under an empty bed hydraulic retention time of 7.8 h, recirculation ratio of 3, and transmembrane pressure of 0.47 bar. The system showed average removals of organics, total nitrogen, and solids as high as 93%, 80%, and 100%, respectively. Rapid nitrification could be achieved and denitrification was performed in the anoxic filter without external carbon supplements. The residual particulate organics and nitrogen in effluent from biofilm process could be also removed successfully through membrane filtration and the removal of total coliform was noticeably improved after membrane filtration. Thus, a system composed of the pre-denitrification biofilm process with ceramic membrane would be a compact and flexible option for advanced post-treatment of municipal wastewater.

  9. Bio-mediated synthesis of TiO2 nanoparticles and its photocatalytic effect on aquatic biofilm.

    PubMed

    Dhandapani, Perumal; Maruthamuthu, Sundram; Rajagopal, Gopalakrishnan

    2012-05-02

    The nano-TiO(2) was synthesized biologically employing Bacillus subtilis (FJ460362). These nanoparticles were characterized by FTIR, TGA-DTA, UV-Visible spectroscopy, XRD and TEM. FTIR and TGA results confirm that the organic impurities were completely removed while calcinating the resultant products. Band gap value was estimated from the UV-Visible spectrum and anatase crystal phase was confirmed by XRD. TEM images reveal that these particles were agglomerated; mostly spherical in shape with an average particle size of 10-30nm. The synthesized nano-TiO(2) particles were coated on glass slides, biofilm were grown and subjected to irradiation of polychromatic light to understand photocatalytic activity in controlling the aquatic biofilm. The bacterial killing process was established by Epi-fluorescence microscopy. The results reveal that biogenic TiO(2) nanomaterial acts as good photocatalyst by the generation of H(2)O(2) in the vicinity of the TiO(2)-biofilm interfaces to suppress the growth of the aquatic biofilm. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. A urea biosensor based on pH-sensitive Sm2TiO5 electrolyte-insulator-semiconductor.

    PubMed

    Pan, Tung-Ming; Huang, Ming-De; Lin, Wan-Ying; Wu, Min-Hsien

    2010-06-11

    A urea biosensor based on pH-sensitive Sm(2)TiO(5) electrolyte-insulator-semiconductor (EIS) has been described. We used X-ray diffraction, Auger electron spectroscopy, and atomic force microscopy to investigate the structural and morphological features of high-k Sm(2)TiO(5) sensing membranes that had been subjected to annealing at different temperatures. The EIS device incorporating a high-k Sm(2)TiO(5) sensing film that had been annealed at 900 degrees C exhibited good sensing characteristics, including a high sensitivity of 60.5 mV/pH (in solutions from pH 2 to 12), a small hysteresis voltage of 2.72 mV (in the pH loop 7-->4-->7-->10-->7), and a low drift rate of 1.15 mV h(-1) (in the buffer solution at pH 7). The Sm(2)TiO(5) EIS device also showed a high selective response towards H(+). This improvement can be attributed to the small number of crystal defects and the large surface roughness. In addition, the urea biosensor based on pH-sensitive EIS incorporating a Sm(2)TiO(5) sensing membrane annealed at 900 degrees C allowed the potentiometric analysis of urea, at concentrations ranging from 0.1 to 32 mM, with a sensitivity of 72.85 mV/purea. Copyright 2010 Elsevier B.V. All rights reserved.

  11. Inhibition of Pseudomonas aeruginosa biofilm formation by 2,2’-bipyridyl, lipoic, kojic and picolinic acids

    PubMed Central

    Çevik, Kübra; Ulusoy, Seyhan

    2015-01-01

    Objective(s): The inhibitory effects of iron chelators, and FeCl3 chelation on biofilm formation and swarming motility were investigated against an opportunistic human pathogen Pseudomonas aeruginosa. Materials and Methods: The inhibitory activity of 2,2’-bipyridyl, lipoic acid, kojic acid and picolinic acid on biofilm formation of P. aeruginosa strain PAO1 and three clinical isolates (P. aeruginosa PAK01, P. aeruginosa PAK02 and P. aeruginosa PAK03) were investigated, based on crystal violet assay, and swarming motility test. Results: The kojic, lipoic and picolinic acid inhibited biofilm formation by 5-33% in all tested P. aeruginosa isolates. When chelated iron was added, biofilm inhibition rates were determined to be 39-57%. Among the tested chelators against P. aeruginosa, lipoic acid (84%) and kojic acid (68%) presented the highest inhibition of swarming motility. This is the first study to report the inhibitory effect of lipoic acid on biofilm formation and swarming motility of P. aeruginosa. Conclusion: It is considered that lipoic and picolinic acids can serve as alternatives for the treatment of the P. aeruginosa infections by inhibiting biofilm formation. PMID:26557964

  12. Prophage insertions in mlrA are not the major obstacle to biofilm formation in Escherichia coli O157:H7

    USDA-ARS?s Scientific Manuscript database

    Background: Although a curli and biofilm expressing phenotype can be induced in Escherichia coli serotype O157:H7, strains examined under laboratory conditions are almost exclusively biofilm deficient and the reason for these deficiencies have remained elusive. CsgD, the central biofilm regulator, i...

  13. Functional Metagenomics of Escherichia coli O157:H7 Interactions with Spinach Indigenous Microorganisms during Biofilm Formation

    PubMed Central

    Carter, Michelle Q.; Xue, Kai; Brandl, Maria T.; Liu, Feifei; Wu, Liyou; Louie, Jacqueline W.; Mandrell, Robert E.; Zhou, Jizhong

    2012-01-01

    The increase in foodborne outbreaks worldwide attributed to fresh fruit and vegetables suggests that produce may serve as an ecological niche for enteric pathogens. Here we examined the interaction of E. coli O157:H7 (EcO157) with spinach leaf indigenous microorganisms during co-colonization and establishment of a mixed biofilm on a stainless steel surface. Stainless steel surface was selected to mimic the surface of produce-processing equipment, where retention of foodborne pathogens such as EcO157 could serve as a potential source for transmission. We observed a positive effect of spinach-associated microbes on the initial attachment of EcO157, but an antagonistic effect on the EcO157 population at the later stage of biofilm formation. Metagenomic analyses of the biofilm community with the GeoChip revealed an extremely diverse community (gene richness, 23409; Shannon-Weiner index H, 9.55). Presence of EcO157 in the mixed biofilm resulted in a significant decrease in the community α-diversity (t test, P<0.05), indicating a putative competition between the pathogen and indigenous spinach microbes. The decrease in the β-diversity of the EcO157-inoculated biofilm at 48 h (ANOVA, P<0.05) suggested a convergent shift in functional composition in response to EcO157 invasion. The success of EcO157 in the mixed biofilm is likely associated with its metabolic potential in utilizing spinach nutrients: the generation time of EcO157 in spinach lysates at 28°C is ∼ 38 min, which is comparable to that in rich broth. The significant decrease in the abundance of many genes involved in carbon, nitrogen, and phosphorus cycling in the EcO157-inoculated biofilms (t test, P<0.05) further support our conclusion that competition for essential macronutrients is likely the primary interaction between the EcO157 and indigenous spinach-biofilm species. PMID:22957052

  14. Antibacterial effect of taurolidine (2%) on established dental plaque biofilm.

    PubMed

    Arweiler, Nicole Birgit; Auschill, Thorsten Mathias; Sculean, Anton

    2012-04-01

    Preliminary data have suggested that taurolidine may bear promising disinfectant properties for the therapy of bacterial infections. However, at present, the potential antibacterial effect of taurolidine on the supragingival plaque biofilm is unknown. To evaluate the antibacterial effect of taurolidine on the supragingival plaque biofilm using the vital fluorescence technique and to compare it with the effect of NaCl and chlorhexidine (CHX), 18 subjects had to refrain from all mechanical and chemical hygiene measures for 24 h. A voluminous supragingival plaque sample was taken from the buccal surfaces of the lower molars and wiped on an objective slide. The sample was then divided into three equal parts and mounted with one of the three test or control preparations (a) NaCl, (b) taurolidine 2% and (c) CHX 0.2%. After a reaction time of 2 min, the test solutions were sucked of. Subsequently, the plaque biofilm was stained with fluorescence dye and vitality of the plaque flora was evaluated under the fluorescence microscope (VF%). Plaque samples treated with NaCl showed a mean VF of 82.42 ± 6.04%. Taurolidine affected mean VF with 47.57 ± 16.60% significantly (p < 0.001, paired t test). The positive control CHX showed the lowest mean VF values (34.41 ± 14.79%; p < 0.001 compared to NaCl, p = 0.017 compared to taurolidine). Taurolidine possesses a significant antibacterial effect on the supragingival plaque biofilm which was, however, not as pronounced as that of CHX.

  15. Xanthomonas citri ssp. citri requires the outer membrane porin OprB for maximal virulence and biofilm formation.

    PubMed

    Ficarra, Florencia A; Grandellis, Carolina; Galván, Estela M; Ielpi, Luis; Feil, Regina; Lunn, John E; Gottig, Natalia; Ottado, Jorgelina

    2017-06-01

    Xanthomonas citri ssp. citri (Xcc) causes canker disease in citrus, and biofilm formation is critical for the disease cycle. OprB (Outer membrane protein B) has been shown previously to be more abundant in Xcc biofilms compared with the planktonic state. In this work, we showed that the loss of OprB in an oprB mutant abolishes bacterial biofilm formation and adherence to the host, and also compromises virulence and efficient epiphytic survival of the bacteria. Moreover, the oprB mutant is impaired in bacterial stress resistance. OprB belongs to a family of carbohydrate transport proteins, and the uptake of glucose is decreased in the mutant strain, indicating that OprB transports glucose. Loss of OprB leads to increased production of xanthan exopolysaccharide, and the carbohydrate intermediates of xanthan biosynthesis are also elevated in the mutant. The xanthan produced by the mutant has a higher viscosity and, unlike wild-type xanthan, completely lacks pyruvylation. Overall, these results suggest that Xcc reprogrammes its carbon metabolism when it senses a shortage of glucose input. The participation of OprB in the process of biofilm formation and virulence, as well as in metabolic changes to redirect the carbon flux, is discussed. Our results demonstrate the importance of environmental nutrient supply and glucose uptake via OprB for Xcc virulence. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  16. One step bioconversion of waste precious metals into Serratia biofilm-immobilized catalyst for Cr(VI) reduction.

    PubMed

    Yong, P; Liu, W; Zhang, Z; Beauregard, D; Johns, M L; Macaskie, L E

    2015-11-01

    For reduction of Cr(VI) the Pd-catalyst is excellent but costly. The objectives were to prove the robustness of a Serratia biofilm as a support for biogenic Pd-nanoparticles and to fabricate effective catalyst from precious metal waste. Nanoparticles (NPs) of palladium were immobilized on polyurethane reticulated foam and polypropylene supports via adhesive biofilm of a Serratia sp. The biofilm adhesion and cohesion strength were unaffected by palladization and catalytic biofilm integrity was also shown by magnetic resonance imaging. Biofilm-Pd and mixed precious metals on biofilm (biofilm-PM) reduced 5 mM Cr(VI) to Cr(III) when immobilized in a flow-through column reactor, at respective flow rates of 9 and 6 ml/h. The lower activity of the latter was attributed to fewer, larger, metal deposits on the bacteria. Activity was lost in each case at pH 7 but was restored by washing with 5 mM citrate solution or by exposure of columns to solution at pH 2, suggesting fouling by Cr(III) hydroxide product at neutral pH. A 'one pot' conversion of precious metal waste into new catalyst for waste decontamination was shown in a continuous flow system based on the use of Serratia biofilm to manufacture and support catalytic Pd-nanoparticles.

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

    NASA Astrophysics Data System (ADS)

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

    2016-04-01

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

  18. Teucrium polium phenylethanol and iridoid glycoside characterization and flavonoid inhibition of biofilm-forming Staphylococcus aureus.

    PubMed

    Elmasri, Wael A; Yang, Tianjiao; Tran, Phat; Hegazy, Mohamed-Elamir F; Hamood, Abdul N; Mechref, Yehia; Paré, Paul W

    2015-01-23

    The chemical composition and biofilm regulation of 15 metabolites from Teucrium polium are reported. Compounds were isolated from a CH2Cl2-MeOH extract of the aerial parts of the plant and included iridoid and phenylethanol glycosides and a monoterpenoid, together with nine known compounds. The structures were elucidated based on standard spectroscopic (UV, (1)H and (13)C NMR), 2D NMR ((1)H-(1)H COSY, HMQC, HMBC, and NOESY), and/or LC-ESIMS/MS data analyses. Inhibition of the biofilm-forming strain Staphylococcus aureus was observed with exposure to compounds 7 and 8.

  19. Acid-triggered membrane insertion of Pseudomonas exotoxin A involves an original mechanism based on pH-regulated tryptophan exposure.

    PubMed

    Méré, Jocelyn; Morlon-Guyot, Juliette; Bonhoure, Anne; Chiche, Laurent; Beaumelle, Bruno

    2005-06-03

    Exposure to low endosomal pH during internalization of Pseudomonas exotoxin A (PE) triggers membrane insertion of its translocation domain. This process is a prerequisite for PE translocation to the cytosol where it inactivates protein synthesis. Although hydrophobic helices enable membrane insertion of related bacterial toxins such as diphtheria toxin, the PE translocation domain is devoid of hydrophobic stretches and the structural features triggering acid-induced membrane insertion of PE are not known. Here we have identified a molecular device that enables PE membrane insertion. This process is promoted by exposure of a key tryptophan residue. At neutral pH, this Trp is buried in a hydrophobic pocket closed by the smallest alpha-helix of the translocation domain. Upon acidification, protonation of the Asp that is the N-cap residue of the helix leads to its destabilization, enabling Trp side chain insertion into the endosome membrane. This tryptophan-based membrane insertion system is surprisingly similar to the membrane-anchoring mechanism of human annexin-V and could be used by other proteins as well.

  20. Antimicrobial activity of a novel bioengineered honey against non-typeable Haemophilus influenzae biofilms: an in vitro study.

    PubMed

    Newby, Rachel S; Dryden, Matthew; Allan, Raymond N; Salib, Rami J

    2018-06-01

    The opportunistic pathogen non-typeable Haemophilus influenzae (NTHi) plays an important role in many chronic respiratory diseases including otitis media, chronic rhinosinusitis, cystic fibrosis and chronic obstructive pulmonary disease. Biofilm formation has been implicated in NTHi colonisation, persistence of infection and recalcitrance towards antimicrobials. There is therefore a pressing need for the development of novel treatment strategies that are effective against NTHi biofilm-associated diseases. SurgihoneyRO is a honey-based product that has been bioengineered to enable the slow release of H 2 O 2 , a reactive oxygen species to which H. influenzae is susceptible. Treatment of established NTHi biofilms with SurgihoneyRO significantly reduced biofilm viability through enhanced H 2 O 2 production and was shown to be more effective than the conventional antibiotic co-amoxiclav. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  1. Addition of ethanol to supercritical carbon dioxide enhances the inactivation of bacterial spores in the biofilm of Bacillus cereus.

    PubMed

    Park, Hyong Seok; Choi, Hee Jung; Kim, Myoung-Dong; Kim, Kyoung Heon

    2013-09-02

    Supercritical carbon dioxide (SC-CO2) was used to inactivate Bacillus cereus spores inside biofilms, which were grown on stainless steel. SC-CO2 treatment was tested using various conditions, such as pressure treatment (10-30 MPa), temperature (35-60°C), and time (10-120 min). B. cereus vegetative cells in the biofilm were completely inactivated by treatment with SC-CO2 at 10 MPa and at 35°C for 5 min. However, SC-CO2 alone did not inactivate spores in biofilm even after the treatment time was extended to 120 min. When ethanol was used as a cosolvent with SC-CO2 in the SC-CO2 treatment using only 2-10 ml of ethanol in 100ml of SC-CO2 vessel for 60-90 min of treatment time at 10 MPa and 60°C, B. cereus spores in the biofilm were found to be completely inactivated in the colony-forming test. We also assessed the viability of SC-CO2-treated bacterial spores and vegetative cells in the biofilm by staining with SYTO 9 and propidium iodide. The membrane integrity of the vegetative cells was completely lost, while the integrity of the membrane was still maintained in most spores. However, when SC-CO2 along with ethanol was used, both vegetative cells and spores lost their membrane integrity, indicating that the use of ethanol as a cosolvent with SC-CO2 is efficient in inactivating the bacterial spores in the biofilm. © 2013.

  2. Membrane-Based Characterization of a Gas Component — A Transient Sensor Theory

    PubMed Central

    Lazik, Detlef

    2014-01-01

    Based on a multi-gas solution-diffusion problem for a dense symmetrical membrane this paper presents a transient theory of a planar, membrane-based sensor cell for measuring gas from both initial conditions: dynamic and thermodynamic equilibrium. Using this theory, the ranges for which previously developed, simpler approaches are valid will be discussed; these approaches are of vital interest for membrane-based gas sensor applications. Finally, a new theoretical approach is introduced to identify varying gas components by arranging sensor cell pairs resulting in a concentration independent gas-specific critical time. Literature data for the N2, O2, Ar, CH4, CO2, H2 and C4H10 diffusion coefficients and solubilities for a polydimethylsiloxane membrane were used to simulate gas specific sensor responses. The results demonstrate the influence of (i) the operational mode; (ii) sensor geometry and (iii) gas matrices (air, Ar) on that critical time. Based on the developed theory the case-specific suitable membrane materials can be determined and both operation and design options for these sensors can be optimized for individual applications. The results of mixing experiments for different gases (O2, CO2) in a gas matrix of air confirmed the theoretical predictions. PMID:24608004

  3. A Nonbactericidal Zinc-Complexing Ligand as a Biofilm Inhibitor: Structure-Guided Contrasting Effects on Staphylococcus aureus Biofilm.

    PubMed

    Kapoor, Vidushi; Rai, Rajanikant; Thiyagarajan, Durairaj; Mukherjee, Sandipan; Das, Gopal; Ramesh, Aiyagari

    2017-08-04

    Zinc-complexing ligands are prospective anti-biofilm agents because of the pivotal role of zinc in the formation of Staphylococcus aureus biofilm. Accordingly, the potential of a thiosemicarbazone (compound C1) and a benzothiazole-based ligand (compound C4) in the prevention of S. aureus biofilm formation was assessed. Compound C1 displayed a bimodal activity, hindering biofilm formation only at low concentrations and promoting biofilm growth at higher concentrations. In the case of C4, a dose-dependent inhibition of S. aureus biofilm growth was observed. Atomic force microscopy analysis suggested that at higher concentrations C1 formed globular aggregates, which perhaps formed a substratum that favored adhesion of cells and biofilm formation. In the case of C4, zinc supplementation experiments validated zinc complexation as a plausible mechanism of inhibition of S. aureus biofilm. Interestingly, C4 was nontoxic to cultured HeLa cells and thus has promise as a therapeutic anti-biofilm agent. The essential understanding of the structure-driven implications of zinc-complexing ligands acquired in this study might assist future screening regimes for identification of potent anti-biofilm agents. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Trypanosoma cruzi H+-ATPase 1 (TcHA1) and 2 (TcHA2) genes complement yeast mutants defective in H+ pumps and encode plasma membrane P-type H+-ATPases with different enzymatic properties.

    PubMed

    Luo, Shuhong; Scott, David A; Docampo, Roberto

    2002-11-15

    Previous studies in Trypanosoma cruzi have shown that intracellular pH homeostasis requires ATP and is affected by H(+)-ATPase inhibitors, indicating a major role for ATP-driven proton pumps in intracellular pH control. In the present study, we report the cloning and sequencing of a pair of genes linked in tandem (TcHA1 and TcHA2) in T. cruzi which encode proteins with homology to fungal and plant P-type proton-pumping ATPases. The genes are expressed at the mRNA level in different developmental stages of T. cruzi: TcHA1 is expressed maximally in epimastigotes, whereas TcHA2 is expressed predominantly in trypomastigotes. The proteins predicted from the nucleotide sequence of the genes have 875 and 917 amino acids and molecular masses of 96.3 and 101.2 kDa, respectively. Full-length TcHA1 and an N-terminal truncated version of TcHA2 complemented a Saccharomyces cerevisiae strain deficient in P-type H(+)-ATPase activity, the proteins localized to the yeast plasma membrane, and ATP-driven proton pumping could be detected in proteoliposomes reconstituted from plasma membrane purified from transfected yeast. The reconstituted proton transport activity was reduced by inhibitors of P-type H(+)-ATPases. C-terminal truncation did not affect complementation of mutant yeast, suggesting the lack of C-terminal autoinhibitory domains in these proteins. ATPase activity in plasma membrane from TcHA1- and (N-terminal truncated) TcHA2-transfected yeast was inhibited to different extents by vanadate, whereas the latter yeast strain was more resistant to extremes of pH, suggesting that the native proteins may serve different functions at different stages in the T. cruzi life cycle.

  5. Effects of Hydrogen Peroxide on Dental Unit Biofilms and Treatment Water Contamination

    PubMed Central

    Lin, Shih-Ming; Svoboda, Kathy K.H.; Giletto, Anthony; Seibert, Jeff; Puttaiah, Raghunath

    2011-01-01

    Objectives: To study effects of various concentrations of hydrogen peroxide on mature waterline biofilms and in controlling planktonic (free-floating) organisms in simulated dental treatment water systems; and to study in vitro the effects of 2%, 3%, and 7% hydrogen peroxide on the removal of mature biofilms and inorganic compounds in dental waterlines. Methods: Four units of an automated dental unit water system simulation device was used for 12 weeks. All units were initially cleaned to control biofilms and inorganic deposits. H2O2 at concentrations of 1%, 2%, 3% was used weekly for periodic cleaning in three treatment group units (units 1, 2 & 3), with 0.05%, 0.15% and 0.25% H2O2 in municipal water used as irrigant respectively. The control unit (unit 4) did not have weekly cleanings and used municipal water as irrigant. Laser Scanning Confocal Microscopy and Scanning Electron Microscopy were used to study deposits on lines, and weekly heterotrophic plate counts done to study effluent water contamination. A 24 hour in vitro challenge test with 7%, 3% and 2% H2O2 on mature biofilms was conducted using harvested waterlines to study biofilm and inorganic deposit removal. Results: Heterotrophic plate counts of effluent water showed that the control unit reached contamination levels in excess of 400,000 CFU/mL while all treatment units showed contamination levels <500 CFU/mL through most of the 12 weeks. All treatment units showed varying levels of biofilm and inorganic deposit control in this short 12 week study. The in vitro challenge test showed although there was biofilm control, there was no eradication even when 7% H2O2 was used for 24 hours. Conclusions: 2% H2O2 used as a periodic cleaner, and diluted to 0.05% in municipal water for irrigation was beneficial in controlling biofilm and planktonic contamination in dental unit water systems. However, to remove well established biofilms, it may take more than 2 months when initial and multiple periodic cleanings

  6. High Specific Selectivity and Membrane-Active Mechanism of Synthetic Cationic Hybrid Antimicrobial Peptides Based on the Peptide FV7

    PubMed Central

    Tan, Tingting; Wu, Di; Li, Weizhong; Zheng, Xin; Li, Weifen; Shan, Anshan

    2017-01-01

    Hybrid peptides integrating different functional domains of peptides have many advantages, such as remarkable antimicrobial activity, lower hemolysis and ideal cell selectivity, compared with natural antimicrobial peptides. FV7 (FRIRVRV-NH2), a consensus amphiphilic sequence was identified as being analogous to host defense peptides. In this study, we designed a series of hybrid peptides FV7-LL-37 (17–29) (FV-LL), FV7-magainin 2 (9–21) (FV-MA) and FV7-cecropin A (1–8) (FV-CE) by combining the FV7 sequence with the small functional sequences LL-37 (17–29) (LL), magainin 2 (9–21) (MA) and cecropin A (1–8) (CE) which all come from well-described natural peptides. The results demonstrated that the synthetic hybrid peptides, in particular FV-LL, had potent antibacterial activities over a wide range of Gram-negative and Gram-positive bacteria with lower hemolytic activity than other peptides. Furthermore, fluorescent spectroscopy indicated that the hybrid peptide FV-LL exhibited marked membrane destruction by inducing outer and inner bacterial membrane permeabilization, while scanning electron microscopy (SEM) and transmission electron microscopy (TEM) demonstrated that FV-LL damaged membrane integrity by disrupting the bacterial membrane. Inhibiting biofilm formation assays also showed that FV-LL had similar anti-biofilm activity compared with the functional peptide sequence FV7. Synthetic cationic hybrid peptides based on FV7 could provide new models for combining different functional domains and demonstrate effective avenues to screen for novel antimicrobial agents. PMID:28178190

  7. Vanadium Redox Flow Batteries Using meta-Polybenzimidazole-Based Membranes of Different Thicknesses.

    PubMed

    Noh, Chanho; Jung, Mina; Henkensmeier, Dirk; Nam, Suk Woo; Kwon, Yongchai

    2017-10-25

    15, 25, and 35 μm thick meta-polybenzimidazole (PBI) membranes are doped with H 2 SO 4 and tested in a vanadium redox flow battery (VRFB). Their performances are compared with those of Nafion membranes. Immersed in 2 M H 2 SO 4 , PBI absorbs about 2 mol of H 2 SO 4 per mole of repeat unit. This results in low conductivity and low voltage efficiency (VE). In ex-situ tests, meta-PBI shows a negligible crossover of V 3+ and V 4+ ions, much lower than that of Nafion. This is due to electrostatic repulsive forces between vanadium cations and positively charged protonated PBI backbones, and the molecular sieving effect of PBI's nanosized pores. It turns out that charge efficiency (CE) of VRFBs using meta-PBI-based membranes is unaffected by or slightly increases with decreasing membrane thickness. Thick meta-PBI membranes require about 100 mV larger potentials to achieve the same charging current as thin meta-PBI membranes. This additional potential may increase side reactions or enable more vanadium ions to overcome the electrostatic energy barrier and to enter the membrane. On this basis, H 2 SO 4 -doped meta-PBI membranes should be thin to achieve high VE and CE. The energy efficiency of 15 μm thick PBI reaches 92%, exceeding that of Nafion 212 and 117 (N212 and N117) at 40 mA cm -2 .

  8. Abundance of the multiheme c-type cytochrome OmcB increases in outer biofilm layers of electrode-grown Geobacter sulfurreducens.

    PubMed

    Stephen, Camille S; LaBelle, Edward V; Brantley, Susan L; Bond, Daniel R

    2014-01-01

    When Geobacter sulfurreducens utilizes an electrode as its electron acceptor, cells embed themselves in a conductive biofilm tens of microns thick. While environmental conditions such as pH or redox potential have been shown to change close to the electrode, less is known about the response of G. sulfurreducens to growth in this biofilm environment. To investigate whether respiratory protein abundance varies with distance from the electrode, antibodies against an outer membrane multiheme cytochrome (OmcB) and cytoplasmic acetate kinase (AckA) were used to determine protein localization in slices spanning ∼25 µm-thick G. sulfurreducens biofilms growing on polished electrodes poised at +0.24 V (vs. Standard Hydrogen Electrode). Slices were immunogold labeled post-fixing, imaged via transmission electron microscopy, and digitally reassembled to create continuous images allowing subcellular location and abundance per cell to be quantified across an entire biofilm. OmcB was predominantly localized on cell membranes, and 3.6-fold more OmcB was detected on cells 10-20 µm distant from the electrode surface compared to inner layers (0-10 µm). In contrast, acetate kinase remained constant throughout the biofilm, and was always associated with the cell interior. This method for detecting proteins in intact conductive biofilms supports a model where the utilization of redox proteins changes with depth.

  9. Integrated combined effects of temperature, pH and sodium chloride concentration on biofilm formation by Salmonella enterica ser. Enteritidis and Typhimurium under low nutrient food-related conditions.

    PubMed

    Iliadis, Ioannis; Daskalopoulou, Aikaterini; Simões, Manuel; Giaouris, Efstathios

    2018-05-01

    Salmonella enterica is a major foodborne bacterial pathogen. This forms biofilms on surfaces and persists, depending on the strain and the environment. The integrative interaction of temperature (T; 13-39 °C), pH (5-8) and sodium chloride (NaCl) concentration (0.5-8.5%) on biofilm formation by two S. enterica strains (ser. Enteritidis and Typhimurium) was here evaluated under low nutrient conditions. This was achieved using response surface methodology to model the combined effect of each factor on the response, through mathematical quadratic fitting of the outcomes of a sequence of designed experiments. These last were executed by incubating stainless steel coupons carrying sessile bacteria, for 24 h, in 1:10 diluted tryptone soya broth, under 15 different combinations of three independent factors (T, pH and NaCl). For each strain, a second order polynomial model, describing the relationship between biofilm formation (log CFU/cm 2 ) and the factors (T, pH and NaCl), was developed using least square regression analysis. Both derived models predicted the combined influences of these factors on biofilm formation, with agreement between predictions and experimental observations (R 2  ≥ 0.96, P ≤ 0.0001). For both strains, the increase of NaCl content restricted their sessile growth, while under low salinity conditions (NaCl < 4%) biofilm formation was favored as pH increased, regardless of T. Interestingly, under low salt content, and depending on the strain, biofilm formation was either favored or hindered by increasing T. Thus, 34.5 and 13 °C were the T predicted to maximize biofilm formation by strains Enteritidis and Typhimurium, respectively, something which was also experimentally verified. To sum, these models can predict the interactive influences of crucial food-related factors on biofilm growth of a significant foodborne pathogen towards the efforts to limit its persistence in food industry. Copyright © 2018 Elsevier Ltd. All rights

  10. Streptococcus gordonii LuxS/autoinducer-2 quorum-sensing system modulates the dual-species biofilm formation with Streptococcus mutans.

    PubMed

    Wang, Xiao; Li, Xiaolan; Ling, Junqi

    2017-07-01

    Dental plaques are mixed-species biofilms that are related to the development of dental caries. Streptococcus mutans (S. mutans) is an important cariogenic bacterium that forms mixed-species biofilms with Streptococcus gordonii (S. gordonii), an early colonizer of the tooth surface. The LuxS/autoinducer-2(AI-2) quorum sensing system is involved in the regulation of mixed-species biofilms, and AI-2 is proposed as a universal signal for the interaction between bacterial species. In this work, a S. gordonii luxS deficient strain was constructed to investigate the effect of the S. gordonii luxS gene on dual-species biofilm formed by S. mutans and S. gordonii. In addition, AI-2 was synthesized in vitro by incubating recombinant LuxS and Pfs enzymes of S. gordonii together. The effect of AI-2 on S. mutans single-species biofilm formation and cariogenic virulence gene expression were also assessed. The results showed that luxS disruption in S. gordonii altered dual-species biofilm formation, architecture, and composition, as well as the susceptibility to chlorhexidine. And the in vitro synthesized AI-2 had a concentration-dependent effect on S. mutans biofilm formation and virulence gene expression. These findings indicate that LuxS/AI-2 quorum-sensing system of S. gordonii plays a role in regulating the dual-species biofilm formation with S. mutans. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Interactions of Cryptosporidium parvum, Giardia lamblia, vaccinal poliovirus type 1, and bacteriophages phiX174 and MS2 with a drinking water biofilm and a wastewater biofilm.

    PubMed

    Helmi, Karim; Skraber, Sylvain; Gantzer, Christophe; Willame, Raphaël; Hoffmann, Lucien; Cauchie, Henry-Michel

    2008-04-01

    Biofilms colonizing surfaces inside drinking water distribution networks may provide a habitat and shelter to pathogenic viruses and parasites. If released from biofilms, these pathogens may disseminate in the water distribution system and cause waterborne diseases. Our study aimed to investigate the interactions of protozoan parasites (Cryptosporidium parvum and Giardia lamblia [oo]cysts) and viruses (vaccinal poliovirus type 1, phiX174, and MS2) with two contrasting biofilms. First, attachment, persistence, and detachment of the protozoan parasites and the viruses were assessed with a drinking water biofilm. This biofilm was allowed to develop inside a rotating annular reactor fed with tap water for 7 months prior to the inoculation. Our results show that viable parasites and infectious viruses attached to the drinking water biofilm within 1 h and persisted within the biofilm. Indeed, infectious viruses were detected in the drinking water biofilm up to 6 days after the inoculation, while viral genome and viable parasites were still detected at day 34, corresponding to the last day of the monitoring period. Since viral genome was detected much longer than infectious particles, our results raise the question of the significance of detecting viral genomes in biofilms. A transfer of viable parasites and viruses from the biofilm to the water phase was observed after the flow velocity was increased but also with a constant laminar flow rate. Similar results regarding parasite and virus attachment and detachment were obtained using a treated wastewater biofilm, suggesting that our observations might be extrapolated to a wide range of environmental biofilms and confirming that biofilms can be considered a potential secondary source of contamination.

  12. Biogenic nanosilver synthesized in Metarhizium robertsii waste mycelium extract – As a modulator of Candida albicans morphogenesis, membrane lipidome and biofilm

    PubMed Central

    Budzyńska, Aleksandra; Bernat, Przemysław

    2018-01-01

    Due to low efficacy of classic antimicrobial drugs, finding new active preparations attracts much attention. In this study an innovative, cost-effective and environmentally friendly method was applied to produce silver nanoparticles (AgNPs) using filamentous fungi Metarhizium robertsii biomass waste. It was shown that these NPs possess prominent antifungal effects against C. albicans, C. glabrata and C. parapsilosis reference strains. Further detailed studies were performed on C. albicans ATCC 90028. AgNPs kill curve (CFU method and esterase-mediated reduction of fluorescein diacetate); fractionally inhibitory concentration index (FICI) with fluconazole (FLC); effect on fungal cell membrane permeability (propidium iodide (PI) staining), membrane lipids profile (HPLC-MS), yeast morphotypes and intracellular reactive oxygen species level (H2DCFDA probe) were investigated. Anti-adhesive and anti-biofilm properties of AgNPs (alone and in combination with FLC) were also tested. Biosafety of AgNPs use was assessed in vitro in cytotoxicity tests against L929 fibroblasts, pulmonary epithelial A549 cell line, and red blood cells. Significant reduction in the viability of yeast cells treated with AgNPs was shown within 6 h. The proportion of C. albicans PI-positive cells increased in a dose and time-dependent manner. Changes in the qualitative and quantitative profile of cell membrane lipids, including significant decline in the quantity of most phospholipid species containing C18:2 and an increase in the amount of phospholipids containing C18:1 acyl species were observed after yeast exposure to AgNPs. CLSM images showed an enhancement in ROS intracellular accumulation in C. albicans treated with biogenic nanosilver. C. albicans transformation from yeast to hyphal forms was also reduced. AgNPs decreased adhesion of yeast to abiotic surfaces, as well as acted synergistically with FLC against sessile population. At fungicidal and fungistatic concentrations, they were non

  13. Preparation of the membrane-permeant biarsenicals, FlAsH-EDT2 and ReAsH-EDT2 for fluorescent labeling of tetracysteine-tagged proteins

    PubMed Central

    Adams, Stephen R.; Tsien, Roger Y.

    2010-01-01

    The membrane-permeant, fluorogenic biarsenicals, FlAsH-EDT2 and ReAsH-EDT2 can be prepared in good yields by a straightforward two-step procedure from the inexpensive precursor dyes, fluorescein and resorufin respectively. Handling of toxic reagents such as arsenic trichloride is minimized so the synthesis can be carried out in a typical chemistry laboratory, typically taking about 2–3 days. A wide range of other biarsenicals reagents and intermediates that also bind to tetracysteine-tagged (CysCysProGlyCysCys) proteins can be prepared similarly using this general procedure. PMID:18772880

  14. The Development of Nitroxide Based Coatings for Biofilm Remediation- 154020

    DTIC Science & Technology

    2017-06-05

    AFRL-AFOSR-JP-TR-2017-0048 The Development of Nitroxide Based Coatings for Biofilm Remediation Kathryn Fairfull-Smith QUEENSLAND UNIVERSITY OF...for Biofilm Remediation - 154020 5a.  CONTRACT NUMBER 5b.  GRANT NUMBER FA2386-15-1-4087 5c.  PROGRAM ELEMENT NUMBER 61102F 6. AUTHOR(S) Kathryn...llisapi.dll Annual Report for AOARD Grant 15IOA087-154020 “The Development of Nitroxide Based Coatings for Biofilm Remediation ” 7th June 2017 PI and Co

  15. Drinking water and biofilm disinfection by Fenton-like reaction.

    PubMed

    Gosselin, F; Madeira, L M; Juhna, T; Block, J C

    2013-10-01

    A Fenton-like disinfection process was conducted with Fenton's reagent (H2O2) at pH 3 or 5 on autochthonous drinking water biofilms grown on corroded or non-corroded pipe material. The biofilm disinfection by Fenton-like oxidation was limited by the low content of iron and copper in the biomass grown on non-corroded plumbing. It was slightly improved by spiking the distribution system with some additional iron source (soluble iron II or ferrihydrite particles appeared as interesting candidates). However successful in situ disinfection of biofilms was only achieved in fully corroded cast iron pipes using H2O2 and adjusting the pH to 5. These new results provide additional support for the use of Fenton's processes for cleaning drinking water distribution systems contaminated with biological agents or organics. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Motility of Pseudomonas aeruginosa contributes to SOS-inducible biofilm formation.

    PubMed

    Chellappa, Shakinah T; Maredia, Reshma; Phipps, Kara; Haskins, William E; Weitao, Tao

    2013-12-01

    DNA-damaging antibiotics such as ciprofloxacin induce biofilm formation and the SOS response through autocleavage of SOS-repressor LexA in Pseudomonas aeruginosa. However, the biofilm-SOS connection remains poorly understood. It was investigated with 96-well and lipid biofilm assays. The effects of ciprofloxacin were examined on biofilm stimulation of the SOS mutant and wild-type strains. The stimulation observed in the wild-type in which SOS was induced was reduced in the mutant in which LexA was made non-cleavable (LexAN) and thus SOS non-inducible. Therefore, the stimulation appeared to involve SOS. The possible mechanisms of inducible biofilm formation were explored by subproteomic analysis of outer membrane fractions extracted from biofilms. The data predicted an inhibitory role of LexA in flagellum function. This premise was tested first by functional and morphological analyses of flagellum-based motility. The flagellum swimming motility decreased in the LexAN strain treated with ciprofloxacin. Second, the motility-biofilm assay was performed, which tested cell migration and biofilm formation. The results showed that wild-type biofilm increased significantly over the LexAN. These results suggest that LexA repression of motility, which is the initial event in biofilm development, contributes to repression of SOS-inducible biofilm formation. Copyright © 2013 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  17. A trait-based approach to bacterial biofilms in soil.

    PubMed

    Lennon, Jay T; Lehmkuhl, Brent K

    2016-09-01

    A trait-based approach focuses on attributes of taxa that influence the structure and function of communities. Biofilm production is a common trait among microorganisms in a wide range of environmental, engineered, and host-associated ecosystems. Here, we used Pseudomonas aeruginosa to link biofilm production to moisture availability, a common stressor for microorganisms in soil. First, we demonstrate that biofilm production is a response trait that influences the desiccation phenotype by increasing survivorship, shifting the niche space, and reducing the minimum water potential needed to sustain a net-positive growth rate (Ψ*). Although the allocation of resources to biofilms is thought to be costly, we found no evidence for a trade-off between fitness and biofilm production along a soil moisture gradient. Second, we demonstrated that biofilm production is an effect trait. Specifically, biofilm production increased water retention in soils that were exposed to a series of drying and rewetting cycles. Although this form of niche construction should affect species interactions, we found no evidence that the benefits of biofilm production were extended to another co-occurring soil bacterium. Together, our results support the view that biofilm production is an important trait that may contribute to the distribution, abundance, and functioning of microorganisms in soils. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  18. Esherichia coli serotype O157:H7 retention on solid surfaces and peroxide resistance is enhanced by dual-strain biofilm formation

    USDA-ARS?s Scientific Manuscript database

    In a previous study we showed that an Escherichia coli O157:H7 strain that was unable to form biofilm could persist in large numbers in dual-strain biofilms formed with an E. coli O-:H4 companion strain. In this study we tested additional companion strains for their ability to retain serotype O157:H...

  19. Effect of biofilm formation, and biocorrosion on denture base fractures.

    PubMed

    Sahin, Cem; Ergin, Alper; Ayyildiz, Simel; Cosgun, Erdal; Uzun, Gulay

    2013-05-01

    The aim of this study was to investigate the destructive effects of biofilm formation and/or biocorrosive activity of 6 different oral microorganisms. Three different heat polymerized acrylic resins (Ivocap Plus, Lucitone 550, QC 20) were used to prepare three different types of samples. Type "A" samples with "V" type notch was used to measure the fracture strength, "B" type to evaluate the surfaces with scanning electron microscopy and "C" type for quantitative biofilm assay. Development and calculation of biofilm covered surfaces on denture base materials were accomplished by SEM and quantitative biofilm assay. According to normality assumptions ANOVA or Kruskal-Wallis was selected for statistical analysis (α=0.05). Significant differences were obtained among the adhesion potential of 6 different microorganisms and there were significant differences among their adhesion onto 3 different denture base materials. Compared to the control groups after contamination with the microorganisms, the three point bending test values of denture base materials decreased significantly (P<.05); microorganisms diffused at least 52% of the denture base surface. The highest median quantitative biofilm value within all the denture base materials was obtained with P. aeruginosa on Lucitone 550. The type of denture base material did not alter the diffusion potential of the microorganisms significantly (P>.05). All the tested microorganisms had destructive effect over the structure and composition of the denture base materials.

  20. Response of plasma membrane H+-ATPase in rice (Oryza sativa) seedlings to simulated acid rain.

    PubMed

    Liang, Chanjuan; Ge, Yuqing; Su, Lei; Bu, Jinjin

    2015-01-01

    Understanding the adaptation of plants to acid rain is important to find feasible approaches to alleviate such damage to plants. We studied effects of acid rain on plasma membrane H(+)-ATPase activity and transcription, intracellular H(+), membrane permeability, photosynthetic efficiency, and relative growth rate during stress and recovery periods. Simulated acid rain at pH 5.5 did not affect plasma membrane H(+)-ATPase activity, intracellular H(+), membrane permeability, photosynthetic efficiency, and relative growth rate. Plasma membrane H(+)-ATPase activity and transcription in leaves treated with acid rain at pH 3.5 was increased to maintain ion homeostasis by transporting excessive H(+) out of cells. Then intracellular H(+) was close to the control after a 5-day recovery, alleviating damage on membrane and sustaining photosynthetic efficiency and growth. Simulated acid rain at pH 2.5 inhibited plasma membrane H(+)-ATPase activity by decreasing the expression of H(+)-ATPase at transcription level, resulting in membrane damage and abnormal intracellular H(+), and reduction in photosynthetic efficiency and relative growth rate. After a 5-day recovery, all parameters in leaves treated with pH 2.5 acid rain show alleviated damage, implying that the increased plasma membrane H(+)-ATPase activity and its high expression were involved in repairing process in acid rain-stressed plants. Our study suggests that plasma membrane H(+)-ATPase can play a role in adaptation to acid rain for rice seedlings.

  1. Atomistic-level study of the interactions between hIAPP protofibrils and membranes: Influence of pH and lipid composition.

    PubMed

    Qian, Zhenyu; Zou, Yu; Zhang, Qingwen; Chen, Peijie; Ma, Buyong; Wei, Guanghong; Nussinov, Ruth

    2018-02-09

    The pathology of type 2 diabetes mellitus is associated with the aggregation of human islet amyloid polypeptide (hIAPP) and aggregation-mediated membrane disruption. The interactions of hIAPP aggregates with lipid membrane, as well as the effects of pH and lipid composition at the atomic level, remain elusive. Herein, using molecular dynamics simulations, we investigate the interactions of hIAPP protofibrillar oligomers with lipids, and the membrane perturbation that they induce, when they are partially inserted in an anionic dipalmitoyl-phosphatidylglycerol (DPPG) membrane or a mixed dipalmitoyl-phosphatidylcholine (DPPC)/DPPG (7:3) lipid bilayer under acidic/neutral pH conditions. We observed that the tilt angles and insertion depths of the hIAPP protofibril are strongly correlated with the pH and lipid composition. At neutral pH, the tilt angle and insertion depth of hIAPP protofibrils at a DPPG bilayer reach ~52° and ~1.62 nm with respect to the membrane surface, while they become ~77° and ~1.75 nm at a mixed DPPC/DPPG membrane. The calculated tilt angle of hIAPP at DPPG membrane is consistent with a recent chiral sum frequency generation spectroscopic study. The acidic pH induces a smaller tilt angle of ~40° and a shallower insertion depth (~1.24 nm) of hIAPP at the DPPG membrane surface, mainly due to protonation of His18 near the turn region. These differences mainly result from a combination of distinct electrostatic, van der Waals, hydrogen bonding and salt-bridge interactions between hIAPP and lipid bilayers. The hIAPP-membrane interaction energy analysis reveals that besides charged residues K1, R11 and H18, aromatic residues Phe15 and Phe23 also exhibit strong interactions with lipid bilayers, revealing the crucial role of aromatic residues in stabilizing the membrane-bound hIAPP protofibrils. hIAPP-membrane interactions disturb the lipid ordering and the local bilayer thickness around the peptides. Our results provide atomic-level information

  2. Biofilm formation and local electrostatic force characteristics of Escherichia coli O157:H7 observed by electrostatic force microscopy

    NASA Astrophysics Data System (ADS)

    Oh, Y. J.; Jo, W.; Yang, Y.; Park, S.

    2007-04-01

    The authors report growth media dependence of electrostatic force characteristics in Escherichia coli O157:H7 biofilm through local measurement by electrostatic force microscopy (EFM). The difference values of electrostatic interaction between the bacterial surface and the abiotic surface show an exponential decay behavior during biofilm development. In the EFM data, the biofilm in the low nutrient media shows a faster decay than the biofilm in the rich media. The surface potential in the bacterial cells was changed from 957to149mV. Local characterization of extracellular materials extracted from the bacteria reveals the progress of the biofilm formation and functional complexities.

  3. Biofouling of reverse-osmosis membranes during tertiary wastewater desalination: microbial community composition.

    PubMed

    Al Ashhab, Ashraf; Herzberg, Moshe; Gillor, Osnat

    2014-03-01

    Reverse-osmosis (RO) desalination is frequently used for the production of high-quality water from tertiary treated wastewater (TTWW). However, the RO desalination process is often hampered by biofouling, including membrane conditioning, microbial adhesion, and biofilm growth. The vast majority of biofilm exploration concentrated on the role of bacteria in biofouling neglecting additional microbial contributors, i.e., fungi and archaea. To better understand the RO biofouling process, bacterial, archaeal and fungal diversity was characterized in a laboratory-scale RO desalination plant exploring the TTWW (RO feed), the RO membrane and the RO feed tube biofilms. We sequenced 77,400 fragments of the ribosome small subunit-encoding gene (16S and 18S rRNA) to identify the microbial community members in these matrices. Our results suggest that the bacterial, archaeal but not fungal community significantly differ from the RO membrane biofouling layer to the feedwater and tube biofilm (P < 0.01). Moreover, the RO membrane supported a more diverse community compared to the communities monitored in the feedwater and the biofilm attached to the RO feedwater tube. The tube biofilm was dominated by Actinobacteria (91.2 ± 4.6%), while the Proteobacteria phylum dominated the feedwater and RO membrane (at relative abundance of 92.3 ± 4.4% and 71.5 ± 8.3%, respectively), albeit comprising different members. The archaea communities were dominated by Crenarchaeota (53.0 ± 6.9%, 32.5 ± 7.2% and 69%, respectively) and Euryarchaeota (43.3 ± 6.3%, 23.2 ± 4.8% and 24%, respectively) in all three matrices, though the communities' composition differed. But the fungal communities composition was similar in all matrices, dominated by Ascomycota (97.6 ± 2.7%). Our results suggest that the RO membrane is a selective surface, supporting unique bacterial, and to a lesser extent archaeal communities, yet it does not select for a fungal community. Copyright © 2013

  4. MPK6 controls H2 O2-induced root elongation by mediating Ca2+ influx across the plasma membrane of root cells in Arabidopsis seedlings.

    PubMed

    Han, Shuan; Fang, Lin; Ren, Xuejian; Wang, Wenle; Jiang, Jing

    2015-01-01

    Mitogen-activated protein kinases (MPKs) play critical roles in signalling and growth, and Ca(2+) and H2 O2 control plant growth processes associated with abscisic acid (ABA). However, it remains unclear how MPKs are involved in H2 O2 - and Ca(2+) -mediated root elongation. Root elongation in seedlings of the loss-of-function mutant Atmpk6 (Arabidopsis thaliana MPK6) was less sensitive to moderate H2 O2 or ABA than that in wild-type (WT) plants. The enhanced elongation was a result of root cell expansion. This effect disappeared when ABA-induced H2 O2 accumulation or the cytosolic Ca(2+) increase were defective. Molecular and biochemical evidence showed that increased expression of the cell wall peroxidase PRX34 in Atmpk6 root cells enhanced apoplastic H2 O2 generation; this promoted a cytosolic Ca(2+) increase and Ca(2+) influx across the plasma membrane. The plasma membrane damage caused by high levels of H2 O2 was ameliorated in a Ca(2+) -dependent manner. These results suggested that there was intensified PRX34-mediated H2 O2 generation in the apoplast and increased Ca(2+) flux into the cytosol of Atmpk6 root cells; that is, the spatial separation of apoplastic H2 O2 from cytosolic Ca(2+) in root cells prevented H2 O2 -induced inhibition of root elongation in Atmpk6 seedlings. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  5. Streptococcus suis Serotype 2 Biofilms Inhibit the Formation of Neutrophil Extracellular Traps.

    PubMed

    Ma, Fang; Yi, Li; Yu, Ningwei; Wang, Guangyu; Ma, Zhe; Lin, Huixing; Fan, Hongjie

    2017-01-01

    Invasive infections caused by Streptococcus suis serotype 2 (SS2) has emerged as a clinical problem in recent years. Neutrophil extracellular traps (NETs) are an important mechanism for the trapping and killing of pathogens that are resistant to phagocytosis. Biofilm formation can protect bacteria from being killed by phagocytes. Until now, there have only been a few studies that focused on the interactions between bacterial biofilms and NETs. SS2 in both a biofilm state and a planktonic cell state were incubated with phagocytes and NETs, and bacterial survival was assessed. DNase I and cytochalasin B were used to degrade NET DNA or suppress phagocytosis, respectively. Extracellular DNA was stained with impermeable fluorescent dye to quantify NET formation. Biofilm formation increased up to 6-fold in the presence of neutrophils, and biofilms were identified in murine tissue. Both planktonic and biofilm cells induced neutrophils chemotaxis to the infection site, with neutrophils increasing by 85.1 and 73.8%, respectively. The bacteria in biofilms were not phagocytized. The bactericidal efficacy of NETs on the biofilms and planktonic cells were equal; however, the biofilm extracellular matrix can inhibit NET release. Although biofilms inhibit NETs release, NETs appear to be an important mechanism to eliminate SS2 biofilms. This knowledge advances the understanding of biofilms and may aid in the development of treatments for persistent infections with a biofilm component.

  6. Antifungal activity of amphotericin B and voriconazole against the biofilms and biofilm-dispersed cells of Candida albicans employing a newly developed in vitro pharmacokinetic model.

    PubMed

    El-Azizi, Mohamed; Farag, Noha; Khardori, Nancy

    2015-04-03

    Candida albicans is a common cause of a variety of superficial and invasive disseminated infections the majority of which are associated with biofilm growth on implanted devices. The aim of the study is to evaluate the activity of amphotericin B and voriconazole against the biofilm and the biofilm-dispersed cells of Candida albicans using a newly developed in vitro pharmacokinetic model which simulates the clinical situation when the antifungal agents are administered intermittently. RPMI medium containing 1-5 X 10(6) CFU/ml of C. albicans was continuously delivered to the device at 30 ml/h for 2 hours. The planktonic cells were removed and biofilms on the catheter were kept under continuous flow of RPMI medium at 10 ml/h. Five doses of amphotericin B or voriconazole were delivered to 2, 5 and 10 day-old biofilms at initial concentrations (2 and 3 μg/ml respectively) that were exponentially diluted. Dispersed cells in effluents from the device were counted and the adherent cells on the catheter were evaluated after 48 h of the last dose. The minimum inhibitory concentration of voriconazole and amphotericin B against the tested isolate was 0.0325 and 0.25 μg/ml respectively. Amphotericin B significantly reduced the dispersion of C. albicans cells from the biofilm. The log10 reduction in the dispersed cells was 2.54-3.54, 2.30-3.55, and 1.94-2.50 following addition of 5 doses of amphotericin B to 2-, 5- and 10-day old biofilms respectively. The number of the viable cells within the biofilm was reduced by 18 (±7.63), 5 and 4% following addition of the 5 doses of amphotericin B to the biofilms respectively. Voriconazole showed no significant effect on the viability of C. albicans within the biofilm. Both antifungal agents failed to eradicate C. albicans biofilm or stop cell dispersion from them and the resistance progressed with maturation of the biofilm. These findings go along with the need for removal of devices in spite of antifungal therapy in patients

  7. Hybrid Membrane/Absorption Process for Post-combustion CO2 Capture

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Li, Shiguang; Shou, S.; Pyrzynski, Travis

    2013-12-31

    with 97% CO2 product purity was achieved throughout the test. Membrane contactor modules have been scaled from bench scale 2-inch diameter by 12-inch long (20 ft2 membrane surface area) modules to 4-inch diameter by 60-inch long pilot scale modules (165 ft2 membrane surface area). Pilot scale modules were tested in an integrated absorption/regeneration system for CO2 capture field tests at a coal-fired power plant (Midwest Generation’s Will County Station located in Romeoville, IL). Absorption and regeneration contactors were constructed utilizing high performance super-hydrophobic, nano-porous PEEK membranes with CO2 gas permeance of 2,000 GPU and a 1,000 GPU, respectively. Field tests using aMDEA solvent achieved greater than 90% CO2 removal in a single stage. The absorption mass transfer coefficient was 1.2 (sec)-1, exceeding the initial target of 1.0 (sec)-1. This mass transfer coefficient is over one order of magnitude greater than that of conventional gas/liquid contacting equipment. The economic evaluation based on field tests data indicates that the CO2 capture cost associated with membrane contactor technology is $54.69 (Yr 2011$)/tonne of CO2 captured when using aMDEA as a solvent. It is projected that the DOE’s 2025 cost goal of $40 (Yr 2011$)/tonne of CO2 captured can be met by decreasing membrane module cost and by utilizing advanced CO2 capture solvents. In the second stage of the field test, an advanced solvent, Hitachi’s H3-1 was utilized. The use of H3-1 solvent increased mass transfer coefficient by 17% as compared to aMDEA solvent. The high mass transfer coefficient of H3-1 solvent combined with much more favorable solvent regeneration requirements, indicate that the projected savings achievable with membrane contactor process can be further improved. H3-1 solvent will be used in the next pilot-scale development phase. The integrated absorption/regeneration process design and high performance membrane contactors developed in the current bench

  8. Effect of the Biofilm Age and Starvation on Acid Tolerance of Biofilm Formed by Streptococcus mutans Isolated from Caries-Active and Caries-Free Adults.

    PubMed

    Jiang, Shan; Chen, Shuai; Zhang, Chengfei; Zhao, Xingfu; Huang, Xiaojing; Cai, Zhiyu

    2017-03-30

    Streptococcus mutans ( S. mutans ) is considered a leading cause of dental caries. The capability of S. mutans to tolerate low pH is essential for its cariogenicity. Aciduricity of S. mutans is linked to its adaptation to environmental stress in oral cavity. This study aimed to investigate the effect of biofilm age and starvation condition on acid tolerance of biofilm formed by S. mutans clinical isolates. S. mutans clinical strains isolated from caries-active (SM593) and caries-free (SM18) adults and a reference strain (ATCC25175) were used for biofilm formation. (1) Both young and mature biofilms were formed and then exposed to pH 3.0 for 30 min with (acid-adapted group) or without (non-adapted group) pre-exposure to pH 5.5 for three hours. (2) The mature biofilms were cultured with phosphate-buffered saline (PBS) (starved group) or TPY (polypeptone-yeast extract) medium (non-starved group) at pH 7.0 for 24 h and then immersed in medium of pH 3.0 for 30 min. Biofilms were analyzed through viability staining and confocal laser scanning microscopy. In all three strains, mature, acid-adapted and starved biofilms showed significantly less destructive structure and more viable bacteria after acid shock than young, non-adapted and non-starved biofilms, respectively (all p < 0.05). Furthermore, in each condition, SM593 biofilm was denser, with a significantly larger number of viable bacteria than that of SM18 and ATCC25175 (all p < 0.05). Findings demonstrated that mature, acid-adapted and starvation might protect biofilms of all three S. mutans strains against acid shock. Additionally, SM593 exhibited greater aciduricity compared to SM18 and ATCC25175, which indicated that the colonization of high cariogenicity of clinical strains may lead to high caries risk in individuals.

  9. Evaluation of process performance, energy consumption and microbiota characterization in a ceramic membrane bioreactor for ex-situ biomethanation of H2 and CO2.

    PubMed

    Alfaro, Natalia; Fdz-Polanco, María; Fdz-Polanco, Fernando; Díaz, Israel

    2018-06-01

    The performance of a pilot ceramic membrane bioreactor for the bioconversion of H 2 and CO 2 to bioCH 4 was evaluated in thermophilic conditions. The loading rate was between 10 and 30 m 3  H 2 /m 3 reactor  d and the system transformed 95% of H 2 fed. The highest methane yield found was 0.22 m 3  CH 4 /m 3  H 2 , close to the maximum stoichiometric value (0.25 m 3  CH 4 /m 3  H 2 ) thus indicating that archaeas employed almost all H 2 transferred to produce CH 4 . k L a value of 268 h -1 was reached at 30 m 3  H 2 /m 3 reactor  d. DGGE and FISH revealed a remarkable archaeas increase related to the selection-effect of H 2 on community composition over time. Methanothermobacter thermautotrophicus was the archaea found with high level of similarity. This study verified the successful application of membrane technology to efficiently transfer H 2 from gas to the liquid phase, the development of a hydrogenotrophic community from a conventional thermophilic sludge and the technical feasibility of the bioconversion. Copyright © 2018 Elsevier Ltd. All rights reserved.

  10. Synergistic disinfection and removal of biofilms by a sequential two-step treatment with ozone followed by hydrogen peroxide.

    PubMed

    Tachikawa, Mariko; Yamanaka, Kenzo

    2014-11-01

    Synergistic disinfection and removal of biofilms by ozone (O3) water in combination with hydrogen peroxide (H2O2) solution was studied by determining disinfection rates and observing changes of the biofilm structure in situ by confocal laser scanning microscopy (CLSM) using an established biofilm of Pseudomonas fluorescence. The sequential treatment with O3, 1.0-1.7 mg/L, followed by H2O2, 0.8-1.1%, showed synergistic disinfection effects, while the reversed treatment, first H2O2 followed by O3, showed only an additive effect. The decrease of synergistic disinfection effect by addition of methanol (CH3OH), a scavenger of hydroxyl radical (OH), into the H2O2 solution suggested generation of hydroxyl radicals on or in the biofilm by the sequential treatment with O3 followed by H2O2. The primary treatment with O3 increased disinfection rates of H2O2 in the secondary treatment, and the increase of O3 concentration enhanced the rates. The cold temperature of O3 water (14 °C and 8 °C) increased the synergistic effect, suggesting the increase of O3 adsorption and hydroxyl radical generation in the biofilm. CLSM observation showed that the sequential treatment, first with O3 followed by H2O2, loosened the cell connections and thinned the extracellular polysaccharides (EPS) in the biofilm. The hydroxyl radical generation in the biofilm may affect the EPS and biofilm structure and may induce effective disinfection with H2O2. This sequential treatment method may suggest a new practical procedure for disinfection and removal of biofilms by inorganic oxidants such as O3 and H2O2. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Biofilm formation and biocides sensitivity of Pseudomonas marginalis isolated from a maple sap collection system.

    PubMed

    Lagacé, L; Jacques, M; Mafu, A A; Roy, D

    2006-10-01

    The susceptibility of planktonic and biofilm cells of Pseudomonas marginalis toward four commonly used biocides at different temperatures (15 and 30 degrees C) and biofilm growth times (24 and 48 h) was assessed. Using the MBEC biofilm device, biofilm production in maple sap was shown to be highly reproducible for each set of conditions tested. Biofilm formation was influenced by growth temperature and time. A temperature of 15 degrees C and incubation time of 24 h yielded fewer CFU per peg and showed fewer adhered cells and typical biofilm structures, based on scanning electron microscopy observations as compared with other conditions. Minimal biofilm eradication concentration values for P. marginalis were significantly greater (P. < 0.001) than were MBCs for planktonic cells and for every biocide tested, with the exception of minimal biofilm eradication concentration values for peracetic acid at 15 degrees C and 24 h. Sodium hypochlorite and peracetic acid sanitizers were able to eliminate P. marginalis biofilms at lower concentrations as compared with hydrogen peroxide- and quaternary ammonium-based sanitizers (P < 0.001). According to the results obtained, sodium hypochlorite and peracetic acid sanitizers would be more appropriate for maple sap collection system sanitation.

  12. Visual detection of cyanide ions by membrane-based nanozyme assay.

    PubMed

    Lien, Chia-Wen; Unnikrishnan, Binesh; Harroun, Scott G; Wang, Chih-Min; Chang, Jia-Yaw; Chang, Huan-Tsung; Huang, Chih-Ching

    2018-04-15

    In this paper, we report a simple one-step synthesis of well-dispersed amorphous cobalt hydroxide/oxide-modified graphene oxide (CoO x H-GO) possessing peroxidase-like catalytic activity, and its application for the detection of H 2 O 2 , glucose, and CN - ions. CoO x H is formed and deposited in situ on the GO surface through the reaction between GO (size ~ 240nm) and Co 2+ in basic solution at room temperature. We investigated the enzyme-mimicking activity of the CoO x H-GO nanohybrid in detail via the H 2 O 2 -mediated oxidation of Amplex Red (AR) to form fluorescent resorufin. The peroxidase-like activity of CoO x H-GO is utilized herein for the quantitation of H 2 O 2 in a wide concentration range, from 100nM to 100μM. When coupled with glucose oxidase (GOD), the AR/CoO x H-GO system can determine glucose level in blood samples. Interestingly, cyanide ions (CN - ) significantly inhibit the catalytic activity of the CoO x H-GO nanohybrid, which allows for the construction of a probe for the detection of CN - in water samples and laboratory wastes. We fabricated a membrane-based CoO x H-GO probe for the visual detection of CN - by preparing a thin film of CoO x H-GO on a positively charged and porous nylon membrane (N + M). The CoO x H-GO/N + M operates on the principle that CN - inhibits the catalytic activity of CoO x H-GO towards the H 2 O 2 -mediated oxidation of AR to form reddish resorufin on the membrane. The intensity of the red color of the membrane decreases with increasing CN - concentration, which can be easily observed with the naked eye at the nanomolar level. This cost-effective sensing system allows for the rapid and simple determination of the concentrations of CN - in complicated wastewater samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Effect of biofilm formation, and biocorrosion on denture base fractures

    PubMed Central

    Ergin, Alper; Ayyildiz, Simel; Cosgun, Erdal; Uzun, Gulay

    2013-01-01

    PURPOSE The aim of this study was to investigate the destructive effects of biofilm formation and/or biocorrosive activity of 6 different oral microorganisms. MATERIALS AND METHODS Three different heat polymerized acrylic resins (Ivocap Plus, Lucitone 550, QC 20) were used to prepare three different types of samples. Type "A" samples with "V" type notch was used to measure the fracture strength, "B" type to evaluate the surfaces with scanning electron microscopy and "C" type for quantitative biofilm assay. Development and calculation of biofilm covered surfaces on denture base materials were accomplished by SEM and quantitative biofilm assay. According to normality assumptions ANOVA or Kruskal-Wallis was selected for statistical analysis (α=0.05). RESULTS Significant differences were obtained among the adhesion potential of 6 different microorganisms and there were significant differences among their adhesion onto 3 different denture base materials. Compared to the control groups after contamination with the microorganisms, the three point bending test values of denture base materials decreased significantly (P<.05); microorganisms diffused at least 52% of the denture base surface. The highest median quantitative biofilm value within all the denture base materials was obtained with P. aeruginosa on Lucitone 550. The type of denture base material did not alter the diffusion potential of the microorganisms significantly (P>.05). CONCLUSION All the tested microorganisms had destructive effect over the structure and composition of the denture base materials. PMID:23755339

  14. Adhesion of CdTe quantum dots on model membranes and internalization into RBL-2H3 cells.

    PubMed

    Zhang, Mengmeng; Wei, Xiaoran; Ding, Lei; Hu, Jingtian; Jiang, Wei

    2017-06-01

    Quantum dots (QDs) have attracted broad attention due to their special optical properties and promising prospect in medical and biological applications. However, the process of QDs on cell membrane is worth further investigations because such process may lead to harmful effects on organisms and also important for QD application. In this study, adhesion of amino- and carboxyl-coated CdTe QDs (A-QDs and C-QDs) on cell membrane and the subsequent internalization are studied using a series of endocytosis-free model membranes, including giant and small unilamellar vesicles, supported lipid bilayers and giant plasma membrane vesicles (GPMVs). The adhered QD amounts on model membranes are quantified by a quartz crystal microbalance. The CdTe QD adhesion on model membranes is governed by electrostatic forces. Positively charged A-QDs adhere on GPMV surface and passively penetrate the plasma membrane via endocytosis-free mechanism, but negatively charged C-QDs cannot. Rat basophilic leukemia (RBL-2H3) cells are exposed to CdTe QDs to monitor the QD internalization process. Both A- and C-QDs are internalized by RBL-2H3 cells mainly via endocytosis. CdTe QDs do not accumulate on the plasma membrane of living cells due to the fast endocytosis and the weakened electrostatic attraction in biological medium, resulting in low chance of passive penetration. The suspended cells after trypsin digestion take more QDs than the adherent cells. A-QDs cause lower cell viability than C-QDs, probably because the approach of positively charged QDs to cells is favored and the smaller aggregates of A-QDs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Assessment of the contamination potentials of some foodborne bacteria in biofilms for food products.

    PubMed

    Adetunji, Victoria O; Adedeji, Adeyemi O; Kwaga, Jacob

    2014-09-01

    To assess biofilms formed by different bacterial strains on glass slides, and changes in biofilm mass and biofilm-associated cell populations after brief contacts between biofilms and either media agar or food products. Two Listeria monocytogenes and Escherichia coli (E. coli) strains and a single Staphylococcus aureus (S. aureus) strain were inoculated separately in tryptic soy broth containing glass coupons incubated for 24, 48 or 72 h at 37 °C. The biofilms formed by individual bacterial strains and biofilm-associated cell populations were determined. Biofilms were subsequently allowed to have brief contacts (1-3 times), through gentle touching, with either agar, meat or soft white cheese (2 cm(3)). Changes in biofilm mass on glass slides and cell populations embedded in biofilms were quantified. A nonpathogenic E. coli formed more biofilms than an E. coli O157:H7 strain. Biofilms formed by S. aureus and Listeria monocytogenes were essentially similar. The biofilm mass increased as incubation time increased within 48 h of incubation and was not positively correlated with cellulose production. Biofilm mass at 48 and 72 h of incubation was not significantly different. More frequent contacts with agar or foods did not remove more biofilms or biofilm-associated cells from glass slides. More S. aureus biofilms were removed followed by Listeria and E. coli biofilms. Mean contamination of agar or food models was 0.00 to 7.65 log CFU/cm(2). Greater contaminations in cell populations were observed with S. aureus and Listeria biofilms. The results provide a clearer assessment of contaminating potential of foods that comes in contact with them. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  16. Interaction of proteins with weak amphoteric charged membrane surfaces: effect of pH.

    PubMed

    Matsumoto, Hidetoshi; Koyama, Yoshiyuki; Tanioka, Akihiko

    2003-08-01

    Weak amphoteric charged membranes were prepared by the graft copolymerization of poly(ethylene glycol) (PEG) derivatives with pendant ionizable groups onto polyethylene (PE) porous membranes. Two types of weak amphoteric charged membranes and two types of weak single charged membranes were prepared. The pH dependence of the protein (fluorescein isothiocyanate-labeled bovine serum albumin, FITC-BSA) adsorption onto the membranes was investigated by fluorescence spectroscopy. The interfacial charge properties of the membranes and protein were also characterized at different pH values by streaming potential and electrophoretic light scattering (ELS) measurements, respectively. The adsorbed amount onto each ionic PEG chain grafted membrane showed a uniform maximum value near the isoelectric point (IEP) of the protein (pH 4.1). On both sides of the IEP (pHs 3.3 and 7.2), the adsorption experiments and zeta (zeta) potential measurements were well correlated: the contribution of electrostatic interaction was dominant for the protein adsorption behavior. In the alkaline condition (pH 10.2), the adsorption experiments contradict the zeta potential measurements. It suggested that the conformational change of protein molecule influenced the adsorption behavior. Finally, these results indicated the potential of controlling the protein-ionic PEG chain interaction on the membrane surfaces by the pH adjustment of the outer solution.

  17. Solution phase and membrane immobilized iron-based free radical reactions: Fundamentals and applications for water treatment

    NASA Astrophysics Data System (ADS)

    Lewis, Scott Romak

    Membrane-based separation processes have been used extensively for drinking water purification, wastewater treatment, and numerous other applications. Reactive membranes synthesized through functionalization of the membrane pores offer enhanced reactivity due to increased surface area at the polymer-solution interface and low diffusion limitations. Oxidative techniques utilizing free radicals have proven effective for both the destruction of toxic organics and non-environmental applications. Most previous work focuses on reactions in the homogeneous phase; however, the immobilization of reactants in membrane pores offers several advantages. The use of polyanions immobilized in a membrane or chelates in solution prevents ferric hydroxide precipitation at near-neutral pH, a common limitation of iron(Fe(II/III))-catalyzed hydrogen peroxide (H 2O2) decomposition. The objectives of this research are to develop a membrane-based platform for the generation of free radicals, degrade toxic organic compounds using this and similar solution-based reactions, degrade toxic organic compounds in droplet form, quantify hydroxyl radical production in these reactions, and develop kinetic models for both processes. In this study, a functionalized membrane containing poly(acrylic acid) (PAA) was used to immobilize iron ions and conduct free radical reactions by permeating H2O2 through the membrane. The membrane's responsive behavior to pH and divalent cations was investigated and modeled. The conversion of Fe(II) to Fe(III) in the membrane and its effect on the decomposition of hydrogen peroxide were monitored and used to develop kinetic models for predicting H2O2 decomposition in these systems. The rate of hydroxyl radical production, and hence contaminant degradation can be varied by changing the residence time, H2O2 concentration, and/or iron loading. Using these membrane-immobilized systems, successful removal of toxic organic compounds, such as pentachlorophenol (PCP), from water

  18. Biofilms produced by Burkholderia cenocepacia: influence of media and solid supports on composition of matrix exopolysaccharides.

    PubMed

    Pellizzoni, Elena; Ravalico, Fabio; Scaini, Denis; Delneri, Ambra; Rizzo, Roberto; Cescutti, Paola

    2016-02-01

    Bacteria usually grow forming biofilms, which are communities of cells embedded in a self-produced dynamic polymeric matrix, characterized by a complex three-dimensional structure. The matrix holds cells together and above a surface, and eventually releases them, resulting in colonization of other surfaces. Although exopolysaccharides (EPOLs) are important components of the matrix, determination of their structure is usually performed on samples produced in non-biofilm conditions, or indirectly through genetic studies. Among the Burkholderia cepacia complex species, Burkholderia cenocepacia is an important pathogen in cystic fibrosis (CF) patients and is generally more aggressive than other species. In the present investigation, B. cenocepacia strain BTS2, a CF isolate, was grown in biofilm mode on glass slides and cellulose membranes, using five growth media, one of which mimics the nutritional content of CF sputum. The structure of the matrix EPOLs was determined by 1H-NMR spectroscopy, while visualization of the biofilms on glass slides was obtained by means of confocal laser microscopy, phase-contrast microscopy and atomic force microscopy. The results confirmed that the type of EPOLs biosynthesized depends both on the medium used and on the type of support, and showed that mucoid conditions do not always lead to significant biofilm production, while bacteria in a non-mucoid state can still form biofilm containing EPOLs.

  19. Biofilm formation by designed co-cultures of Caldicellulosiruptor species as a means to improve hydrogen productivity.

    PubMed

    Pawar, Sudhanshu S; Vongkumpeang, Thitiwut; Grey, Carl; van Niel, Ed Wj

    2015-01-01

    Caldicellulosiruptor species have gained a reputation as being among the best microorganisms to produce hydrogen (H2) due to possession of a combination of appropriate features. However, due to their low volumetric H2 productivities (Q H2), Caldicellulosiruptor species cannot be considered for any viable biohydrogen production process yet. In this study, we evaluate biofilm forming potential of pure and co-cultures of Caldicellulosiruptor saccharolyticus and Caldicellulosiruptor owensensis in continuously stirred tank reactors (CSTR) and up-flow anaerobic (UA) reactors. We also evaluate biofilms as a means to retain biomass in the reactor and its influence on Q H2. Moreover, we explore the factors influencing the formation of biofilm. Co-cultures of C. saccharolyticus and C. owensensis form substantially more biofilm than formed by C. owensensis alone. Biofilms improved substrate conversion in both of the reactor systems, but improved the Q H2 only in the UA reactor. When grown in the presence of each other's culture supernatant, both C. saccharolyticus and C. owensensis were positively influenced on their individual growth and H2 production. Unlike the CSTR, UA reactors allowed retention of C. saccharolyticus and C. owensensis when subjected to very high substrate loading rates. In the UA reactor, maximum Q H2 (approximately 20 mmol · L(-1)  · h(-1)) was obtained only with granular sludge as the carrier material. In the CSTR, stirring negatively affected biofilm formation. Whereas, a clear correlation was observed between elevated (>40 μM) intracellular levels of the secondary messenger bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) and biofilm formation. In co-cultures C. saccharolyticus fortified the trade of biofilm formation by C. owensensis, which was mediated by elevated levels of c-di-GMP in C. owensensis. These biofilms were effective in retaining biomass of both species in the reactor and improving Q H2 in a UA reactor using

  20. Preparation of a novel pH optical sensor using orange (II) based on agarose membrane as support.

    PubMed

    Heydari, Rouhollah; Hosseini, Mohammad; Amraei, Ahmadreza; Mohammadzadeh, Ali

    2016-04-01

    A novel and cost effective optical pH sensor was prepared using covalent immobilization of orange (II) indicator on the agarose membrane as solid support. The fabricated optical sensor was fixed into a sample holder of a spectrophotometer instrument for pH monitoring. Variables affecting sensor performance including pH of dye bonding to agarose membrane and dye concentration were optimized. The sensor responds to the pH changes in the range of 3.0-10.0 with a response time of 2.0 min and appropriate reproducibility (RSD ≤ 0.9%). No significant variation was observed on sensor response after increasing the ionic strength in the range of 0.0-0.5M of sodium chloride. Determination of pH using the proposed optical sensor is quick, simple, inexpensive, selective and sensitive in the pH range of 3.0-10.0. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Metabolic Microenvironmental Control by Photosynthetic Biofilms under Changing Macroenvironmental Temperature and pH Conditions▿ †

    PubMed Central

    Bissett, Andrew; Reimer, Andreas; de Beer, Dirk; Shiraishi, Fumito; Arp, Gernot

    2008-01-01

    Ex situ microelectrode experiments, using cyanobacterial biofilms from karst water creeks, were conducted under various pH, temperature, and constant-alkalinity conditions to investigate the effects of changing environmental parameters on cyanobacterial photosynthesis-induced calcification. Microenvironmental chemical conditions around calcifying sites were controlled by metabolic activity over a wide range of photosynthesis and respiration rates, with little influence from overlying water conditions. Regardless of overlying water pH levels (from 7.8 to 8.9), pH at the biofilm surface was approximately 9.4 in the light and 7.8 in the dark. The same trend was observed at various temperatures (4°C and 17°C). Biological processes control the calcium carbonate saturation state (Ω) in these and similar systems and are able to maintain Ω at approximately constant levels over relatively wide environmental fluctuations. Temperature did, however, have an effect on calcification rate. Calcium flux in this system is limited by its diffusion coefficient, resulting in a higher calcium flux (calcification and dissolution) at higher temperatures, despite the constant, biologically mediated pH. The ability of biological systems to mitigate the effects of environmental perturbation is an important factor that must be considered when attempting to predict the effects of increased atmospheric partial CO2 pressure on processes such as calcification and in interpreting microfossils in the fossil record. PMID:18689512

  2. Disinfection efficacy over yeast biofilms of juice processing industries.

    PubMed

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

    2018-03-01

    Membrane separation systems represent a hot - spot for biofilm formation in juice industries. Sodium hypochlorite (NaOCl) has been traditionally the disinfectant of choice; however, its effectiveness over well-established biofilms is limited. In this work the study of biofilm formation on ultrafiltration membranes was proposed. The effectiveness of cleaning and disinfection procedures commonly used in juice industry was tested on the removal and killing of cells. The species used (Rhodotorula mucilaginosa, Candida krusei, Candida kefyr and Candida tropicalis) were isolated from ultrafiltration modules of a clarified apple juice industry. Industrial concentrations of NaOCl (200mgCL∙L -1 ) showed to be effective against planktonic cultures with >4 log reductions, whereas their overall efficiency against adhered cells was smaller. Recovery of viable cell counts to initial numbers was evidenced regardless of the time of colonization. The topography of the surface showed to have an impact on the efficiency of the disinfectant, presenting membranes smaller log reductions than stainless steel (~1.09-1.53logCFU). At 200mgCl∙L -1 only membrane's cross flow recovery was reached with no long-term effect over the attached cells. The overall results demonstrated the recalcitrance of these biofilms to typical cleaning and disinfection process which may confer them with a selective advantage. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Antimicrobial blue light inactivation of biofilms formed by clinical isolates of multidrug-resistant microorganisms

    NASA Astrophysics Data System (ADS)

    Ferrer-Espada, Raquel; Fang, Yanyan; Dai, Tianhong

    2018-02-01

    Antibiotic resistance is one of the most serious threats to public health. It is estimated that at least 23,000 people die each year in the USA as a direct result of antibiotic-resistant infections. In addition, many antibiotic-resistant microorganisms develop biofilms, surface-associated microbial communities that are extremely resistant to antibiotics and the immune system. A light-based approach, antimicrobial blue light (aBL), has attracted increasing attention due to its intrinsic antimicrobial effect without the involvement of exogenous photosensitizers. In this study, we investigated the effectiveness of this non-antibiotic approach against biofilms formed by multidrug-resistant (MDR) microorganisms. MDR Acinetobacter baumannii, Escherichia coli, Candida albicans, and Pseudomonas aeruginosa biofilms were grown either in 96-well microtiter plates for 24 h or in a CDC biofilm reactor for 48 h, and then exposed to aBL at 405 nm emitted from a light-emitting diode (LED). We demonstrated that, for the biofilms grown in the CDC biofilm reactor, approximately 1.88 log10 CFU reduction was achieved in A. baumannii, 2.78 log10 CFU in E. coli and 3.18 log10 CFU in P. aeruginosa after 162 J/cm2 , 576 J/cm2 and 500 J/cm2 aBL were delivered, respectively. For the biofilms formed in the 96-well microtiter plates, 5.67 and 2.46 log10 CFU reduction was observed in P. aeruginosa and C. albicans polymicrobial biofilm after an exposure of 216 J/cm2 . In conclusion, aBL is potentially an alternative non-antibiotic approach against MDR biofilm-related infections. Future studies are warranted to investigate other important MDR microorganisms, the mechanism of action of aBL, and aBL efficacy in vivo.

  4. Inhibition of biofilm formation in Bacillus subtilis by new halogenated furanones.

    PubMed

    Kayumov, Airat R; Khakimullina, Elvina N; Sharafutdinov, Irshad S; Trizna, Elena Y; Latypova, Lilia Z; Thi Lien, Hoang; Margulis, Anna B; Bogachev, Mikhail I; Kurbangalieva, Almira R

    2015-05-01

    Gram-positive bacteria can cause various infections including hospital-acquired infections. While in the biofilm, the resistance of bacteria to both antibiotics and the human immune system is increased causing difficulties in the treatment. Bacillus subtilis, a non-pathogenic Gram-positive bacterium, is widely used as a model organism for studying biofilm formation. Here we investigated the effect of novel synthesized chloro- and bromo-containing 2(5H)-furanones on biofilm formation by B. subtilis. Mucobromic acid (3,4-dibromo-5-hydroxy-2(5H)-furanone) and the two derivatives of mucochloric acid (3,4-dichloro-5-hydroxy-2(5H)-furanone)-F8 and F12-were found to inhibit the growth and to efficiently prevent biofilm formation by B. subtilis. Along with the low production of polysaccharide matrix and repression of the eps operon, strong repression of biofilm-related yqxM also occurred in the presence of furanones. Therefore, our data confirm that furanones affect significantly the regulatory pathway(s) leading to biofilm formation. We propose that the global regulator, Spo0A, is one of the potential putative cellular targets for these compounds.

  5. Genetic and physiological effects of noncoherent visible light combined with hydrogen peroxide on Streptococcus mutans in biofilm.

    PubMed

    Steinberg, Doron; Moreinos, Daniel; Featherstone, John; Shemesh, Moshe; Feuerstein, Osnat

    2008-07-01

    Oral biofilms are associated with the most common infections of the oral cavity. Bacteria embedded in the biofilms are less sensitive to antibacterial agents than planktonic bacteria are. Recently, an antibacterial synergic effect of noncoherent blue light and hydrogen peroxide (H(2)O(2)) on planktonic Streptococcus mutans was demonstrated. In this study, we tested the effect of a combination of light and H(2)O(2) on the vitality and gene expression of S. mutans embedded in biofilm. Biofilms of S. mutans were exposed to visible light (wavelengths, 400 to 500 nm) for 30 or 60 s (equivalent to 34 or 68 J/cm(2)) in the presence of 3 to 300 mM H(2)O(2). The antibacterial effect was assessed by microbial counts of each treated sample compared with that of the control. The effect of light combined with H(2)O(2) on the different layers of the biofilm was evaluated by confocal laser scanning microscopy. Gene expression was determined by real-time reverse transcription-PCR. Our results show that noncoherent light, in combination with H(2)O(2), has a synergistic antibacterial effect through all of the layers of the biofilm. Furthermore, this treatment was more effective against bacteria in biofilm than against planktonic bacteria. The combined light and H(2)O(2) treatment up-regulated the expression of several genes such as gtfB, brp, smu630, and comDE but did not affect relA and ftf. The ability of noncoherent visible light in combination with H(2)O(2) to affect bacteria in deep layers of the biofilm suggests that this treatment may be applied in biofilm-related diseases as a minimally invasive antibacterial procedure.

  6. Design and field application of a UV-LED based optical fiber biofilm sensor.

    PubMed

    Fischer, Matthias; Wahl, Martin; Friedrichs, Gernot

    2012-03-15

    Detecting changes in the formation dynamics of biofilms stemming from bacteria and unicellular microorganisms in their natural environment is of prime interest for biological, ecological as well as anti-fouling technology research. We developed a robust optical fiber-based biofilm sensor ready to be applied in natural aquatic environments for on-line, in situ and non-destructive monitoring of large-area biofilms. The device is based on the detection of the natural fluorescence of microorganisms constituting the biofilm. Basically, the intrinsic fluorescence of the amino acid tryptophan is excited at a wavelength of λ=280 nm and detected at λ=350 nm utilising a numerically optimized sensor head equipped with a UV-LED light source and optical fiber bundles for efficient fluorescence light collection. Calibration was carried out with tryptophan solutions and two characteristic marine bacteria strains revealing linear signal response, satisfactory background suppression, wide dynamic range, and an experimental detection limit of 4 × 10(3)cells/cm(2). Successful field experiments in the Baltic Sea accomplished over a period of twenty-one days provided for the first time continuous observation of biofilm formation dynamics in a natural habitat. Starting from the first adhering bacteria, the measurement yielded the characteristic three phases of biofilm formation up to a fully developed biofilm. The sensor system holds potential for applications in aquatic sciences including deep sea research and, after further miniaturisation, in the industrial and biomedical field. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Dynamic response performance of proton exchange membrane fuel cell stack with Pt/C-RuO2·xH2O electrode

    NASA Astrophysics Data System (ADS)

    Lu, Lu; Xu, Hongfeng; Zhao, Hong; Sun, Xin; Dong, Yiming; Ren, Ruiming

    2013-11-01

    The dynamic response performance of a proton exchange membrane fuel cell (PEMFC) significantly affects its durability and reliability. Thus, the improvement of the dynamic performance of PEMFC has become the key for prolonging the PEMFC life in fuel cell vehicle applications. In this study, RuO2·xH2O is prepared by sol-gel method, and then sprayed onto catalyst layers to promote PEMFC dynamic response performance. The prepared RuO2·xH2O is characterized by TEM, which shows that the average particle size of RuO2·xH2O is 8 nm and that the particulates are uniformly distributed. A 10-cell stack is assembled using membrane electrode assembly (MEA) with and without RuO2·xH2O. This stack is studied under various loading cycles and operating conditions, including different air stoichiometries, relative humidities, and loading degrees. Results show that the steady-state performance of the MEA with RuO2·xH2O is better than that in the MEA without RuO2·xH2O with a decreasing relative humidity from 80% to 20%. A slower and more unstable dynamic response of the MEA without RuO2·xH2O is observed as air stoichiometry and relative humidity decrease as well as the loading increase. Thus, RuO2·xH2O improves the dynamic response performance, indicating that RuO2·xH2O can buffer the voltage undershoot, improve the stability, and prolong the lifetime of the PEMFC stack.

  8. Antibacterial photocatalytic activity of different crystalline TiO2 phases in oral multispecies biofilm.

    PubMed

    Pantaroto, Heloisa N; Ricomini-Filho, Antonio P; Bertolini, Martinna M; Dias da Silva, José Humberto; Azevedo Neto, Nilton F; Sukotjo, Cortino; Rangel, Elidiane C; Barão, Valentim A R

    2018-07-01

    Titanium dioxide (TiO 2 ) incorporation in biomaterials is a promising technology due to its photocatalytic and antibacterial activities. However, the antibacterial potential of different TiO 2 crystalline structures on a multispecies oral biofilm remains unknown. We hypothesized that the different crystalline TiO 2 phases present different photocatalytic and antibacterial activities. Three crystalline TiO 2 films were deposited by magnetron sputtering on commercially pure titanium (cpTi), in order to obtain four groups: (1) machined cpTi (control); (2) A-TiO 2 (anatase); (3) M-TiO 2 (mixture of anatase and rutile); (4) R-TiO 2 (rutile). The morphology, crystalline phase, chemical composition, hardness, elastic modulus and surface free energy of the surfaces were evaluated. The photocatalytic potential was assessed by methylene blue degradation assay. The antibacterial activity was evaluated on relevant oral bacteria, by using a multispecies biofilm (Streptococcus sanguinis, Actinomyces naeslundii and Fusobacterium nucleatum) formed on the treated titanium surfaces (16.5h) followed by UV-A light exposure (1h) to generate reactive oxygen species production. All TiO 2 films presented around 300nm thickness and improved the hardness and elastic modulus of cpTi surfaces (p<0.05). A-TiO 2 and M-TiO 2 films presented superior photocatalytic activity than R-TiO 2 (p<0.05). M-TiO 2 revealed the greatest antibacterial activity followed by A-TiO 2 (≈99.9% and 99% of bacterial reduction, respectively) (p<0.001 vs. control). R-TiO 2 had no antibacterial activity (p>0.05 vs. control). This study brings new insights on the development of extra oral protocols for the photocatalytic activity of TiO 2 in oral biofilm-associated disease. Anatase and mixture-TiO 2 showed antibacterial activity on this oral bacterial biofilm, being promising surface coatings for dental implant components. Copyright © 2018 The Academy of Dental Materials. All rights reserved.

  9. An improved protocol for harvesting Bacillus subtilis colony biofilms.

    PubMed

    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. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nguyen, Hung D.; Cao, Bin; Mishra, Bhoopesh

    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 (<500-1000more » 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.« less

  11. Breastfeeding, Dental Biofilm Acidogenicity, and Early Childhood Caries.

    PubMed

    Neves, Pierre A M; Ribeiro, Cecília Claudia Costa; Tenuta, Livia Maria Andaló; Leitão, Tarcísio J; Monteiro-Neto, Valério; Nunes, Ana Margarida M; Cury, Jaime Aparecido

    2016-01-01

    This study evaluated the acidogenicity of human milk by the dental biofilms of children with and without early childhood caries (ECC). Biofilms of 16 children (7 with ECC; 9 caries free) were exposed to human milk or 10% sucrose solution in the crossover design, and the biofilm pH was determined. Breastfeeding did not provoke a decrease in biofilm pH, irrespective of the children's caries status, whereas sucrose decreased the pH for both groups. In addition, higher x0394;pH5min (pH variation occurring at 5 min) was observed in the biofilms of ECC children (p < 0.05). The results suggest that breastfeeding may not contribute to ECC. © 2016 S. Karger AG, Basel.

  12. The effect of hydrodynamic conditions on the phenotype of Pseudomonas fluorescens biofilms.

    PubMed

    Simões, Manuel; Pereira, Maria O; Sillankorva, Sanna; Azeredo, Joana; Vieira, Maria J

    2007-01-01

    This study investigated the phenotypic characteristics of monoculture P. fluorescens biofilms grown under turbulent and laminar flow, using flow cells reactors with stainless steel substrata. The cellular physiology and the overall biofilm activity, structure and composition were characterized, and compared, within hydrodynamically distinct conditions. The results indicate that turbulent flow-generated biofilm cells were significantly less extensive, with decreased metabolic activity and a lower protein and polysaccharides composition per cell than those from laminar flow-generated biofilms. The effect of flow regime did not cause significantly different outer membrane protein expression. From the analysis of biofilm activity, structure and composition, turbulent flow-generated biofilms were metabolically more active, had twice more mass per cm(2), and higher cellular density and protein content (mainly cellular) than laminar flow-generated biofilms. Conversely, laminar flow-generated biofilms presented higher total and matrix polysaccharide contents. Direct visualisation and scanning electron microscopy analysis showed that these different flows generate structurally different biofilms, corroborating the quantitative results. The combination of applied methods provided useful information regarding a broad spectrum of biofilm parameters, which can contribute to control and model biofilm processes.

  13. Predicting adhesion and biofilm formation boundaries on stainless steel surfaces by five Salmonella enterica strains belonging to different serovars as a function of pH, temperature and NaCl concentration.

    PubMed

    Moraes, Juliana O; Cruz, Ellen A; Souza, Enio G F; Oliveira, Tereza C M; Alvarenga, Verônica O; Peña, Wilmer E L; Sant'Ana, Anderson S; Magnani, Marciane

    2018-05-26

    This study aimed to assess the capability of 97 epidemic S. enterica strains belonging to 18 serovars to form biofilm. Five strains characterized as strong biofilm-producers, belonging to distinct serovars (S. Enteritidis 132, S. Infantis 176, S. Typhimurium 177, S. Heidelberg 281 and S. Corvallis 297) were assayed for adhesion/biofilm formation on stainless steel surfaces. The experiments were conducted in different combinations of NaCl (0, 2, 4, 5, 6, 8 and 10% w/v), pH (4, 5, 6 and 7) and temperatures (8 °C, 12 °C, 20 °C and 35 °C). Only adhesion was assumed to occur when S. enterica counts were ≥3 and <5 log CFU/cm 2 , whereas biofilm formation was defined as when the counts were ≥5 log CFU/cm 2 . The binary responses were used to develop models to predict the probability of adhesion/biofilm formation on stainless steel surfaces by five strains belonging to different S. enterica serovars. A total of 99% (96/97) of the tested S. enterica strains were characterized as biofilm-producers in the microtiter plate assays. The ability to form biofilm varied (P < 0.05) within and among the different serovars. Among the biofilm-producers, 21% (20/96), 45% (43/96), and 35% (34/96) were weak, moderate and strong biofilm-producers, respectively. The capability for adhesion/biofilm formation on stainless steel surfaces under the experimental conditions studied varied among the strains studied, and distinct secondary models were obtained to describe the behavior of the five S. enterica tested. All strains showed adhesion at pH 4 up to 4% of NaCl and at 20 °C and 35 °C. The probability of adhesion decreased when NaCl concentrations were >8% and at 8 °C, as well as in pH values ≤ 5 and NaCl concentrations > 6%, for all tested strains. At pH 7 and 6, biofilm formation for S. Enteritidis, S. Infantis, S. Typhimurium, S. Heidelberg was observed up to 6% of NaCl at 35 °C and 20 °C. The predicted boundaries for adhesion were pH

  14. Effect of temperature, pH, and water activity on biofilm formation by Salmonella enterica enteritidis PT4 on stainless steel surfaces as indicated by the bead vortexing method and conductance measurements.

    PubMed

    Giaouris, E; Chorianopoulos, N; Nychas, G J E

    2005-10-01

    An assay was developed in an effort to elucidate the effect of important environmental parameters (temperature, pH, and water activity [aw]) on Salmonella Enteritidis biofilm formation on stainless steel surfaces. To achieve this, a modified microbiological technique used for biofilm studying (the bead vortexing method) and a rapid method based on conductivity measurements were used. The ability of the microorganism to generate biofilm on the stainless surfaces was studied at three temperatures (5, 20, and 37 degrees C), four pH values (4.5, 5.5, 6.5, and 7.4), and four aw values (0.5, 1.5, 5.5, and 10.5% NaCl). Results obtained by the bead vortexing method show that maximum numbers of adherent bacteria per square centimeter (106 CFU/cm2) were attained in 6 days at 20 degrees C. Biofilm formation after 7 days of incubation at 20 degrees C was found to be independent of the pH value. In addition, the high concentration of sodium chloride (10.5% NaCl, aw = 0.94) clearly inhibited the adherence of cells to the coupons. Conductance measurements were used as a supplementary tool to measure indirectly the attachment and biofilm formation of bacterial cells on stainless steel surfaces via their metabolic activity (i.e., changes in the conductance of the growth medium due to microbial growth or metabolism). Results obtained by conductance measurements corresponded well to those of the bead vortexing method. Furthermore, we were able to detect cells that remained attached on the metal surfaces even after vortexing via their metabolic activity. The results, except for demonstrating environmental-dependent Salmonella Enteritidis biofilm formation, indicated that traditional vortexing with beads did not remove completely biofilm cells from stainless steel; hence, conductance measurements seem to provide a more sensitive test capable to detect down to one single viable organism.

  15. Calcium transcriptionally regulates the biofilm machinery of Xylella fastidiosa to promote continued biofilm development in batch cultures.

    PubMed

    Parker, Jennifer K; Chen, Hongyu; McCarty, Sara E; Liu, Lawrence Y; De La Fuente, Leonardo

    2016-05-01

    The functions of calcium (Ca) in bacteria are less characterized than in eukaryotes, where its role has been studied extensively. The plant-pathogenic bacterium Xylella fastidiosa has several virulence features that are enhanced by increased Ca concentrations, including biofilm formation. However, the specific mechanisms driving modulation of this feature are unclear. Characterization of biofilm formation over time showed that 4 mM Ca supplementation produced denser biofilms that were still developing at 96 h, while biofilm in non-supplemented media had reached the dispersal stage by 72 h. To identify changes in global gene expression in X. fastidiosa grown in supplemental Ca, RNA-Seq of batch culture biofilm cells was conducted at three 24-h time intervals. Results indicate that a variety of genes are differentially expressed in response to Ca, including genes related to attachment, motility, exopolysaccharide synthesis, biofilm formation, peptidoglycan synthesis, regulatory functions, iron homeostasis, and phages. Collectively, results demonstrate that Ca supplementation induces a transcriptional response that promotes continued biofilm development, while biofilm cells in nonsupplemented media are driven towards dispersion of cells from the biofilm structure. These results have important implications for disease progression in planta, where xylem sap is the source of Ca and other nutrients for X. fastidiosa. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  16. Dynamic Dispersal of Surface Layer Biofilm Induced by Nanosized TiO2 Based on Surface Plasmon Resonance and Waveguide.

    PubMed

    Zhang, Peng; Guo, Jin-Song; Yan, Peng; Chen, You-Peng; Wang, Wei; Dai, You-Zhi; Fang, Fang; Wang, Gui-Xue; Shen, Yu

    2018-05-01

    Pollutant degradation is present mainly in the surface layer of biofilms, and the surface layer is the most vulnerable to impairment by toxic pollutants. In this work, the effects of nanosized TiO 2 (n-TiO 2 ) on the average thicknesses of Bacillus subtilis biofilm and on bacterial attachment on different surfaces were investigated. The binding mechanism of n-TiO 2 to the cell surface was also probed. The results revealed that n-TiO 2 caused biofilm dispersal and the thicknesses decreased by 2.0 to 2.6 μm after several hours of exposure. The attachment abilities of bacteria with extracellular polymeric substances (EPS) on hydrophilic surfaces were significantly reduced by 31% and 81% under 10 and 100 mg/liter of n-TiO 2 , respectively, whereas those of bacteria without EPS were significantly reduced by 43% and 87%, respectively. The attachment abilities of bacteria with and without EPS on hydrophobic surfaces were significantly reduced by 50% and 56%, respectively, under 100 mg/liter of n-TiO 2 The results demonstrated that biofilm dispersal can be attributed to the changes in the cell surface structure and the reduction of microbial attachment ability. IMPORTANCE Nanoparticles can penetrate into the outer layer of biofilm in a relatively short period and can bind onto EPS and bacterial surfaces. The current work probed the effects of nanosized TiO 2 (n-TiO 2 ) on biofilm thickness, bacterial migration, and surface properties of the cell in the early stage using the surface plasmon resonance waveguide mode. The results demonstrated that n-TiO 2 decreased the adhesive ability of both cell and EPS and induced bacterial migration and biofilm detachment in several hours. The decreased adhesive ability of microbes and EPS worked against microbial aggregation, reducing the effluent quality in the biological wastewater treatment process. Copyright © 2018 American Society for Microbiology.

  17. Hibiscus sabdariffa extract inhibits in vitro biofilm formation capacity of Candida albicans isolated from recurrent urinary tract infections.

    PubMed

    Alshami, Issam; Alharbi, Ahmed E

    2014-02-01

    To explore the prevention of recurrent candiduria using natural based approaches and to study the antimicrobial effect of Hibiscus sabdariffa (H. sabdariffa) extract and the biofilm forming capacity of Candida albicans strains in the present of the H. sabdariffa extract. In this particular study, six strains of fluconazole resistant Candida albicans isolated from recurrent candiduria were used. The susceptibility of fungal isolates, time-kill curves and biofilm forming capacity in the present of the H. sabdariffa extract were determined. Various levels minimum inhibitory concentration of the extract were observed against all the isolates. Minimum inhibitory concentration values ranged from 0.5 to 2.0 mg/mL. Time-kill experiment demonstrated that the effect was fungistatic. The biofilm inhibition assay results showed that H. sabdariffa extract inhibited biofilm production of all the isolates. The results of the study support the potential effect of H. sabdariffa extract for preventing recurrent candiduria and emphasize the significance of the plant extract approach as a potential antifungal agent.

  18. Halogenated Phenazines that Potently Eradicate Biofilms, MRSA Persister Cells in Non-Biofilm Cultures, and Mycobacterium tuberculosis.

    PubMed

    Garrison, Aaron T; Abouelhassan, Yasmeen; Kallifidas, Dimitris; Bai, Fang; Ukhanova, Maria; Mai, Volker; Jin, Shouguang; Luesch, Hendrik; Huigens, Robert W

    2015-12-01

    Conventional antibiotics are ineffective against non-replicating bacteria (for example, bacteria within biofilms). We report a series of halogenated phenazines (HP), inspired by marine antibiotic 1, that targets persistent bacteria. HP 14 demonstrated the most potent biofilm eradication activities to date against MRSA, MRSE, and VRE biofilms (MBEC = 0.2-12.5 μM), as well as the effective killing of MRSA persister cells in non-biofilm cultures. Frontline MRSA treatments, vancomycin and daptomycin, were unable to eradicate MRSA biofilms or non-biofilm persisters alongside 14. HP 13 displayed potent antibacterial activity against slow-growing M. tuberculosis (MIC = 3.13 μM), the leading cause of death by bacterial infection around the world. HP analogues effectively target persistent bacteria through a mechanism that is non-toxic to mammalian cells and could have a significant impact on treatments for chronic bacterial infections. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Acidic pH Strongly Enhances In Vitro Biofilm Formation by a Subset of Hypervirulent ST-17 Streptococcus agalactiae Strains

    PubMed Central

    D'Urzo, Nunzia; Pezzicoli, Alfredo; De Cesare, Virginia; Pinto, Vittoria; Margarit, Immaculada; Telford, John Laird; Maione, Domenico

    2014-01-01

    Streptococcus agalactiae, also known as group B Streptococcus (GBS), is a primary colonizer of the anogenital mucosa of up to 40% of healthy women and an important cause of invasive neonatal infections worldwide. Among the 10 known capsular serotypes, GBS type III accounts for 30 to 76% of the cases of neonatal meningitis. In recent years, the ability of GBS to form biofilm attracted attention for its possible role in fitness and virulence. Here, a new in vitro biofilm formation protocol was developed to guarantee more stringent conditions, to better discriminate between strong-, low-, and non-biofilm-forming strains, and to facilitate interpretation of data. This protocol was used to screen the biofilm-forming abilities of 366 GBS clinical isolates from pregnant women and from neonatal infections of different serotypes in relation to medium composition and pH. The results identified a subset of isolates of serotypes III and V that formed strong biofilms under acidic conditions. Importantly, the best biofilm formers belonged to serotype III hypervirulent clone ST-17. Moreover, the abilities of proteinase K to strongly inhibit biofilm formation and to disaggregate mature biofilms suggested that proteins play an essential role in promoting GBS biofilm initiation and contribute to biofilm structural stability. PMID:24487536

  20. Resistance of Legionella pneumophila serotype 1 biofilms to chlorine-based disinfection.

    PubMed

    Cooper, I R; Hanlon, G W

    2010-02-01

    The presence of Legionella spp. in potable water systems is a major concern to municipal water providers and consumers alike. Despite the inclusion of chlorine in potable supplies and frequent chlorination cycles, the bacterium is a recalcitrant human pathogen capable of causing incidents of Legionnaires' disease, Pontiac fever and community-acquired pneumonia in humans. Using two materials routinely employed for the delivery of potable water as a substratum, copper and stainless steel, the development of Legionella pneumophila biofilms and their response to chlorination was monitored over a three-day and a three-month period, respectively. Preliminary in vitro studies using broth and sterile tap water as culture media indicated that the bacterium was capable of surviving in low numbers for 28 days in the presence of chlorine. Subsequently, biofilms were grown for three days, one month and two months, respectively, on stainless steel and copper sections, which are widely used for the conveyance of potable water. Immediately after exposure to 50mg/L chlorine for 1h, the biofilms yielded no recoverable colonies, but colonies did reappear in low numbers over the following days. Despite chlorination at 50mg/L for 1h, both one- and two-month-old L. pneumophila biofilms were able to survive this treatment and to continue to grow, ultimately exceeding 1x10(6)cfu per disc. This research provides an insight into the resistance afforded to L. pneumophila against high levels of chlorine by the formation of biofilms and has implications for the delivery of potable water.

  1. Enhanced Shewanella biofilm promotes bioelectricity generation.

    PubMed

    Liu, Ting; Yu, Yang-Yang; Deng, Xiao-Peng; Ng, Chun Kiat; Cao, Bin; Wang, Jing-Yuan; Rice, Scott A; Kjelleberg, Staffan; Song, Hao

    2015-10-01

    Electroactive biofilms play essential roles in determining the power output of microbial fuel cells (MFCs). To engineer the electroactive biofilm formation of Shewanella oneidensis MR-1, a model exoelectrogen, we herein heterologously overexpressed a c-di-GMP biosynthesis gene ydeH in S. oneidensis MR-1, constructing a mutant strain in which the expression of ydeH is under the control of IPTG-inducible promoter, and a strain in which ydeH is under the control of a constitutive promoter. Such engineered Shewanella strains had significantly enhanced biofilm formation and bioelectricity generation. The MFCs inoculated with these engineered strains accomplished a maximum power density of 167.6 ± 3.6 mW/m(2) , which was ∼ 2.8 times of that achieved by the wild-type MR-1 (61.0 ± 1.9 mW/m(2) ). In addition, the engineered strains in the bioelectrochemical system at poised potential of 0.2 V vs. saturated calomel electrode (SCE) generated a stable current density of 1100 mA/m(2) , ∼ 3.4 times of that by wild-type MR-1 (320 mA/m(2) ). © 2015 Wiley Periodicals, Inc.

  2. Streptokinase Treatment Reverses Biofilm-Associated Antibiotic Resistance in Staphylococcus aureus.

    PubMed

    Jørgensen, Nis Pedersen; Zobek, Natalia; Dreier, Cindy; Haaber, Jakob; Ingmer, Hanne; Larsen, Ole Halfdan; Meyer, Rikke L

    2016-09-20

    Biofilms formed by Staphylococcus aureus is a serious complication to the use of medical implants. A central part of the pathogenesis relies on S. aureus' ability to adhere to host extracellular matrix proteins, which adsorb to medical implants and stimulate biofilm formation. Being coagulase positive, S. aureus furthermore induces formation of fibrin fibers from fibrinogen in the blood. Consequently, we hypothesized that fibrin is a key component of the extracellular matrix of S. aureus biofilms under in vivo conditions, and that the recalcitrance of biofilm infections can be overcome by combining antibiotic treatment with a fibrinolytic drug. We quantified S. aureus USA300 biofilms grown on peg-lids in brain heart infusion (BHI) broth with 0%-50% human plasma. Young (2 h) and mature (24 h) biofilms were then treated with streptokinase to determine if this lead to dispersal. Then, the minimal biofilm eradication concentration (MBEC) of 24 h old biofilms was measured for vancomycin and daptomycin alone or in combination with 10 µg/mL rifampicin in the presence or absence of streptokinase in the antibiotic treatment step. Finally, biofilms were visualized by confocal laser scanning microscopy. Addition of human plasma stimulated biofilm formation in BHI in a dose-dependent manner, and biofilms could be partially dispersed by streptokinase. The biofilms could be eradicated with physiologically relevant concentrations of streptokinase in combination with rifampicin and vancomycin or daptomycin, which are commonly used antibiotics for treatment of S. aureus infections. Fibronolytic drugs have been used to treat thromboembolic events for decades, and our findings suggest that their use against biofilm infections has the potential to improve the efficacy of antibiotics in treatment of S. aureus biofilm infections.

  3. Evidence for biofilm acid neutralization by baking soda.

    PubMed

    Zero, Domenick T

    2017-11-01

    The generating of acids from the microbial metabolism of dietary sugars and the subsequent decrease in biofilm pH below the pH at which tooth mineral begins to demineralize (critical pH) are the key elements of the dental caries process. Caries preventive strategies that rapidly neutralize biofilm acids can prevent demineralization and favor remineralization and may help prevent the development of sugar-induced dysbiosis that shifts the biofilm toward increased cariogenic potential. Although the neutralizing ability of sodium bicarbonate (baking soda) has been known for many years, its anticaries potential as an additive to fluoride dentifrice has received only limited investigation. There is evidence that baking soda rapidly can reverse the biofilm pH decrease after a sugar challenge; however, the timing of when it is used in relation to a dietary sugar exposure is critical in that the sooner its used the greater the benefit in preventing a sustained biofilm pH decrease and subsequent demineralization. Furthermore, the effectiveness of baking soda in elevating biofilm pH appears to depend on concentration. Thus, the concentration of baking soda in marketed dentifrice products, which ranges from 10% to 65%, may affect their biofilm pH neutralizing performance. People with hyposalivation particularly may benefit from using fluoride dentifrice containing baking soda because of their diminished ability to clear dietary sugars and buffer biofilm acids. Although promising, there is the need for more evidence that strategies that modify the oral ecology, such as baking soda, can alter the cariogenic (acidogenic and aciduric) properties of biofilm microorganisms. The acid neutralization of dental biofilm by using fluoride dentifrice that contains baking soda has potential for helping counteract modern high-sugar diets by rapidly neutralizing biofilm-generated acid, especially in people with hyposalivation. Copyright © 2017 American Dental Association. Published by

  4. Current and future trends for biofilm reactors for fermentation processes.

    PubMed

    Ercan, Duygu; Demirci, Ali

    2015-03-01

    Biofilms in the environment can both cause detrimental and beneficial effects. However, their use in bioreactors provides many advantages including lesser tendencies to develop membrane fouling and lower required capital costs, their higher biomass density and operation stability, contribution to resistance of microorganisms, etc. Biofilm formation occurs naturally by the attachment of microbial cells to the support without use of any chemicals agent in biofilm reactors. Biofilm reactors have been studied and commercially used for waste water treatment and bench and pilot-scale production of value-added products in the past decades. It is important to understand the fundamentals of biofilm formation, physical and chemical properties of a biofilm matrix to run the biofilm reactor at optimum conditions. This review includes the principles of biofilm formation; properties of a biofilm matrix and their roles in the biofilm formation; factors that improve the biofilm formation, such as support materials; advantages and disadvantages of biofilm reactors; and industrial applications of biofilm reactors.

  5. In situ hydrogen utilization for high fraction acetate production in mixed culture hollow-fiber membrane biofilm reactor.

    PubMed

    Zhang, Fang; Ding, Jing; Shen, Nan; Zhang, Yan; Ding, Zhaowei; Dai, Kun; Zeng, Raymond J

    2013-12-01

    Syngas fermentation is a promising route for resource recovery. Acetate is an important industrial chemical product and also an attractive precursor for liquid biofuels production. This study demonstrated high fraction acetate production from syngas (H₂ and CO₂) in a hollow-fiber membrane biofilm reactor, in which the hydrogen utilizing efficiency reached 100% during the operational period. The maximum concentration of acetate in batch mode was 12.5 g/L, while the acetate concentration in continuous mode with a hydraulic retention time of 9 days was 3.6 ± 0.1 g/L. Since butyrate concentration was rather low and below 0.1 g/L, the acetate fraction was higher than 99% in both batch and continuous modes. Microbial community analysis showed that the biofilm was dominated by Clostridium spp., such as Clostridium ljungdahlii and Clostridium drakei, the percentage of which was 70.5%. This study demonstrates a potential technology for the in situ utilization of syngas and valuable chemical production.

  6. Genetic and Physiological Effects of Noncoherent Visible Light Combined with Hydrogen Peroxide on Streptococcus mutans in Biofilm

    PubMed Central

    Steinberg, Doron; Moreinos, Daniel; Featherstone, John; Shemesh, Moshe; Feuerstein, Osnat

    2008-01-01

    Oral biofilms are associated with the most common infections of the oral cavity. Bacteria embedded in the biofilms are less sensitive to antibacterial agents than planktonic bacteria are. Recently, an antibacterial synergic effect of noncoherent blue light and hydrogen peroxide (H2O2) on planktonic Streptococcus mutans was demonstrated. In this study, we tested the effect of a combination of light and H2O2 on the vitality and gene expression of S. mutans embedded in biofilm. Biofilms of S. mutans were exposed to visible light (wavelengths, 400 to 500 nm) for 30 or 60 s (equivalent to 34 or 68 J/cm2) in the presence of 3 to 300 mM H2O2. The antibacterial effect was assessed by microbial counts of each treated sample compared with that of the control. The effect of light combined with H2O2 on the different layers of the biofilm was evaluated by confocal laser scanning microscopy. Gene expression was determined by real-time reverse transcription-PCR. Our results show that noncoherent light, in combination with H2O2, has a synergistic antibacterial effect through all of the layers of the biofilm. Furthermore, this treatment was more effective against bacteria in biofilm than against planktonic bacteria. The combined light and H2O2 treatment up-regulated the expression of several genes such as gtfB, brp, smu630, and comDE but did not affect relA and ftf. The ability of noncoherent visible light in combination with H2O2 to affect bacteria in deep layers of the biofilm suggests that this treatment may be applied in biofilm-related diseases as a minimally invasive antibacterial procedure. PMID:18316516

  7. Vigabatrin transport across the human intestinal epithelial (Caco-2) brush-border membrane is via the H+-coupled amino-acid transporter hPAT1

    PubMed Central

    Abbot, Emily L; Grenade, Danielle S; Kennedy, David J; Gatfield, Kelly M; Thwaites, David T

    2005-01-01

    The aim of this investigation was to determine if the human proton-coupled amino-acid transporter 1 (hPAT1 or SLC36A1) is responsible for the intestinal uptake of the orally-administered antiepileptic agent 4-amino-5-hexanoic acid (vigabatrin). The Caco-2 cell line was used as a model of the human small intestinal epithelium. Competition experiments demonstrate that [3H]GABA uptake across the apical membrane was inhibited by vigabatrin and the GABA analogues trans-4-aminocrotonic acid (TACA) and guvacine, whereas 1-(aminomethyl)cyclohexaneacetic acid (gabapentin) had no affect. Experiments with 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF)-loaded Caco-2 cells demonstrate that apical exposure to vigabatrin and TACA induce comparable levels of intracellular acidification (due to H+/amino-acid symport) to that generated by GABA, suggesting that they are substrates for a H+-coupled absorptive transporter such as hPAT1. In hPAT1 and mPAT1-expressing Xenopus laevis oocytes [3H]GABA uptake was inhibited by vigabatrin, TACA and guvacine, whereas gabapentin failed to inhibit [3H]GABA uptake. In Na+-free conditions, vigabatrin and TACA evoked similar current responses (due to H+/amino-acid symport) in hPAT1-expressing oocytes under voltage-clamp conditions to that induced by GABA (whereas no current was observed in water-injected oocytes) consistent with the ability of these GABA analogues to inhibit [3H]GABA uptake. This study demonstrates that hPAT1 is the carrier responsible for the uptake of vigabatrin across the brush-border membrane of the small intestine and emphasises the therapeutic potential of hPAT1 as a delivery route for orally administered, clinically significant GABA-related compounds. PMID:16331283

  8. Antimicrobial activity of essential oils against Staphylococcus aureus biofilms.

    PubMed

    Vázquez-Sánchez, Daniel; Cabo, Marta L; Rodríguez-Herrera, Juan J

    2015-12-01

    The present study was aimed to evaluate the potential of essential oils to remove the foodborne pathogen Staphylococcus aureus from food-processing facilities. The effectiveness of 19 essential oils against planktonic cells of S. aureus was firstly assessed by minimal inhibitory concentration. Planktonic cells showed a wide variability in resistance to essential oils, with thyme oil as the most effective, followed by lemongrass oil and then vetiver oil. The eight essential oils most effective against planktonic cells were subsequently tested against 48-h-old biofilms formed on stainless steel. All essential oils reduced significantly (p < 0.01) the number of viable biofilm cells, but none of them could remove biofilms completely. Thyme and patchouli oils were the most effective, but high concentrations were needed to achieve logarithmic reductions over 4 log CFU/cm(2) after 30 min exposure. Alternatively, the use of sub-lethal doses of thyme oil allowed to slow down biofilm formation and to enhance the efficiency of thyme oil and benzalkonium chloride against biofilms. However, some cellular adaptation to thyme oil was detected. Therefore, essential oil-based treatments should be based on the rotation and combination of different essential oils or with other biocides to prevent the emergence of antimicrobial-resistant strains. © The Author(s) 2014.

  9. Antimicrobial photodynamic therapy (aPDT) induction of biofilm matrix architectural and bioadhesive modifications.

    PubMed

    Mang, Thomas; Rogers, Stephen; Keinan, David; Honma, Kiyonobu; Baier, Robert

    2016-03-01

    Dental implants are commonly used today for the treatment of partially and fully edentulous patients. Despite the high success rate they are not resistant to complications and failure due to a variety of problems including peri-implantitis or peri-mucositis due to bacterial biofilm formation on the implant surface. The use of non-surgical and surgical treatment procedure to promote healing in cases with peri-implantitis have limited efficacy. Here we studied the ability of photodynamic therapy to destroy a known bacterial pathogen and the extracellular matrix architecture of biofilm attached to titanium plates and germanium prisms. Titanium plates or germanium prisms were incubated for 24h with Fusobacterium nucleatum a fusiform, gram-negative bacterium was used to enable biofilm formation. Photodynamic therapy was carried out by incubating the biofilm samples on each substrata with porfimer sodium. Treatment was carried out using a diode laser at 630nm, 150mW/cm(2) for light doses ranging from 25-100J/cm(2). Evaluation of killing efficacy was done by counting colony forming units compared to controls. Multiple attenuated internal reflection-infrared spectroscopy (MAIR-IR) and SEM were used to analyze the samples pre and post PDT for validation. F. nucleatum was significantly reduced in a dose dependent manner by treatment with PDT. Changes in biofilm components and strength of bioadhesion were examined with MAIR-IR following jet impingement using calibrated water jets. SEM demonstrates significant morphological alterations in the bacteria, consistent with damage associated with exposure to reactive oxygen species. The results are indicative that aPDT is a method that can be used to eradicate micro-organisms associated with biofilm in peri-implantitis on relevant substrata. Data shows that the slime layer of the biofilm is removed and that further methods need to be employed to completely remove weakened or destroyed biofilm matrix components. Reactive oxygen

  10. Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation1[OPEN

    PubMed Central

    Okumura, Masaki; Inoue, Shin-ichiro; Kuwata, Keiko

    2016-01-01

    Plant plasma membrane H+-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H+-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha. However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H+-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H+-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H+-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H+-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H+-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H+-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H+-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. PMID:27016447

  11. Bacteriophage Lysin CF-301, a Potent Antistaphylococcal Biofilm Agent

    PubMed Central

    Khan, Babar K.; Raz, Assaf; Rotolo, Jimmy A.; Wittekind, Michael

    2017-01-01

    ABSTRACT Biofilms pose a unique therapeutic challenge because of the antibiotic tolerance of constituent bacteria. Treatments for biofilm-based infections represent a major unmet medical need, requiring novel agents to eradicate mature biofilms. Our objective was to evaluate bacteriophage lysin CF-301 as a new agent to target Staphylococcus aureus biofilms. We used minimum biofilm-eradicating concentration (MBEC) assays on 95 S. aureus strains to obtain a 90% MBEC (MBEC90) value of ≤0.25 μg/ml for CF-301. Mature biofilms of coagulase-negative staphylococci, Streptococcus pyogenes, and Streptococcus agalactiae were also sensitive to disruption, with MBEC90 values ranging from 0.25 to 8 μg/ml. The potency of CF-301 was demonstrated against S. aureus biofilms formed on polystyrene, glass, surgical mesh, and catheters. In catheters, CF-301 removed all biofilm within 1 h and killed all released bacteria by 6 h. Mixed-species biofilms, formed by S. aureus and Staphylococcus epidermidis on several surfaces, were removed by CF-301, as were S. aureus biofilms either enriched for small-colony variants (SCVs) or grown in human synovial fluid. The antibacterial activity of CF-301 was further demonstrated against S. aureus persister cells in exponential-phase and stationary-phase populations. Finally, the antibiofilm activity of CF-301 was greatly improved in combinations with the cell wall hydrolase lysostaphin when tested against a range of S. aureus strains. In all, the data show that CF-301 is highly effective at disrupting biofilms and killing biofilm bacteria, and, as such, it may be an efficient new agent for treating staphylococcal infections with a biofilm component. PMID:28461319

  12. Bacteriophage Lysin CF-301, a Potent Antistaphylococcal Biofilm Agent.

    PubMed

    Schuch, Raymond; Khan, Babar K; Raz, Assaf; Rotolo, Jimmy A; Wittekind, Michael

    2017-07-01

    Biofilms pose a unique therapeutic challenge because of the antibiotic tolerance of constituent bacteria. Treatments for biofilm-based infections represent a major unmet medical need, requiring novel agents to eradicate mature biofilms. Our objective was to evaluate bacteriophage lysin CF-301 as a new agent to target Staphylococcus aureus biofilms. We used minimum biofilm-eradicating concentration (MBEC) assays on 95 S. aureus strains to obtain a 90% MBEC (MBEC 90 ) value of ≤0.25 μg/ml for CF-301. Mature biofilms of coagulase-negative staphylococci, Streptococcus pyogenes , and Streptococcus agalactiae were also sensitive to disruption, with MBEC 90 values ranging from 0.25 to 8 μg/ml. The potency of CF-301 was demonstrated against S. aureus biofilms formed on polystyrene, glass, surgical mesh, and catheters. In catheters, CF-301 removed all biofilm within 1 h and killed all released bacteria by 6 h. Mixed-species biofilms, formed by S. aureus and Staphylococcus epidermidis on several surfaces, were removed by CF-301, as were S. aureus biofilms either enriched for small-colony variants (SCVs) or grown in human synovial fluid. The antibacterial activity of CF-301 was further demonstrated against S. aureus persister cells in exponential-phase and stationary-phase populations. Finally, the antibiofilm activity of CF-301 was greatly improved in combinations with the cell wall hydrolase lysostaphin when tested against a range of S. aureus strains. In all, the data show that CF-301 is highly effective at disrupting biofilms and killing biofilm bacteria, and, as such, it may be an efficient new agent for treating staphylococcal infections with a biofilm component. Copyright © 2017 American Society for Microbiology.

  13. Glutathione-Disrupted Biofilms of Clinical Pseudomonas aeruginosa Strains Exhibit an Enhanced Antibiotic Effect and a Novel Biofilm Transcriptome.

    PubMed

    Klare, William; Das, Theerthankar; Ibugo, Amaye; Buckle, Edwina; Manefield, Mike; Manos, Jim

    2016-08-01

    Pseudomonas aeruginosa infections result in high morbidity and mortality rates for individuals with cystic fibrosis (CF), with premature death often occurring. These infections are complicated by the formation of biofilms in the sputum. Antibiotic therapy is stymied by antibiotic resistance of the biofilm matrix, making novel antibiofilm strategies highly desirable. Within P. aeruginosa biofilms, the redox factor pyocyanin enhances biofilm integrity by intercalating with extracellular DNA. The antioxidant glutathione (GSH) reacts with pyocyanin, disrupting intercalation. This study investigated GSH disruption by assaying the physiological effects of GSH and DNase I on biofilms of clinical CF isolates grown in CF artificial sputum medium (ASMDM+). Confocal scanning laser microscopy showed that 2 mM GSH, alone or combined with DNase I, significantly disrupted immature (24-h) biofilms of Australian epidemic strain (AES) isogens AES-1R and AES-1M. GSH alone greatly disrupted mature (72-h) AES-1R biofilms, resulting in significant differential expression of 587 genes, as indicated by RNA-sequencing (RNA-seq) analysis. Upregulated systems included cyclic diguanylate and pyoverdine biosynthesis, the type VI secretion system, nitrate metabolism, and translational machinery. Biofilm disruption with GSH revealed a cellular physiology distinct from those of mature and dispersed biofilms. RNA-seq results were validated by biochemical and quantitative PCR assays. Biofilms of a range of CF isolates disrupted with GSH and DNase I were significantly more susceptible to ciprofloxacin, and increased antibiotic effectiveness was achieved by increasing the GSH concentration. This study demonstrated that GSH, alone or with DNase I, represents an effective antibiofilm treatment when combined with appropriate antibiotics, pending in vivo studies. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  14. Glutathione-Disrupted Biofilms of Clinical Pseudomonas aeruginosa Strains Exhibit an Enhanced Antibiotic Effect and a Novel Biofilm Transcriptome

    PubMed Central

    Das, Theerthankar; Ibugo, Amaye; Buckle, Edwina; Manefield, Mike; Manos, Jim

    2016-01-01

    Pseudomonas aeruginosa infections result in high morbidity and mortality rates for individuals with cystic fibrosis (CF), with premature death often occurring. These infections are complicated by the formation of biofilms in the sputum. Antibiotic therapy is stymied by antibiotic resistance of the biofilm matrix, making novel antibiofilm strategies highly desirable. Within P. aeruginosa biofilms, the redox factor pyocyanin enhances biofilm integrity by intercalating with extracellular DNA. The antioxidant glutathione (GSH) reacts with pyocyanin, disrupting intercalation. This study investigated GSH disruption by assaying the physiological effects of GSH and DNase I on biofilms of clinical CF isolates grown in CF artificial sputum medium (ASMDM+). Confocal scanning laser microscopy showed that 2 mM GSH, alone or combined with DNase I, significantly disrupted immature (24-h) biofilms of Australian epidemic strain (AES) isogens AES-1R and AES-1M. GSH alone greatly disrupted mature (72-h) AES-1R biofilms, resulting in significant differential expression of 587 genes, as indicated by RNA-sequencing (RNA-seq) analysis. Upregulated systems included cyclic diguanylate and pyoverdine biosynthesis, the type VI secretion system, nitrate metabolism, and translational machinery. Biofilm disruption with GSH revealed a cellular physiology distinct from those of mature and dispersed biofilms. RNA-seq results were validated by biochemical and quantitative PCR assays. Biofilms of a range of CF isolates disrupted with GSH and DNase I were significantly more susceptible to ciprofloxacin, and increased antibiotic effectiveness was achieved by increasing the GSH concentration. This study demonstrated that GSH, alone or with DNase I, represents an effective antibiofilm treatment when combined with appropriate antibiotics, pending in vivo studies. PMID:27161630

  15. Mitigation of Membrane Biofouling in MBR Using a Cellulolytic Bacterium, Undibacterium sp. DM-1, Isolated from Activated Sludge.

    PubMed

    Nahm, Chang Hyun; Lee, Seonki; Lee, Sang Hyun; Lee, Kibaek; Lee, Jaewoo; Kwon, Hyeokpil; Choo, Kwang-Ho; Lee, Jung-Kee; Jang, Jae Young; Lee, Chung-Hak; Park, Pyung-Kyu

    2017-03-28

    Biofilm formation on the membrane surface results in the loss of permeability in membrane bioreactors (MBRs) for wastewater treatment. Studies have revealed that cellulose is not only produced by a number of bacterial species but also plays a key role during formation of their biofilm. Hence, in this study, cellulase was introduced to a MBR as a cellulose-induced biofilm control strategy. For practical application of cellulase to MBR, a cellulolytic ( i.e ., cellulase-producing) bacterium, Undibacterium sp. DM-1, was isolated from a lab-scale MBR for wastewater treatment. Prior to its application to MBR, it was confirmed that the cell-free supernatant of DM-1 was capable of inhibiting biofilm formation and of detaching the mature biofilm of activated sludge and cellulose-producing bacteria. This suggested that cellulase could be an effective anti-biofouling agent for MBRs used in wastewater treatment. Undibacterium sp. DM-1-entrapping beads ( i.e ., cellulolytic-beads) were applied to a continuous MBR to mitigate membrane biofouling 2.2-fold, compared with an MBR with vacant-beads as a control. Subsequent analysis of the cellulose content in the biofilm formed on the membrane surface revealed that this mitigation was associated with an approximately 30% reduction in cellulose by cellulolytic-beads in MBR.

  16. Anti-biofilm properties of the antimicrobial peptide temporin 1Tb and its ability, in combination with EDTA, to eradicate Staphylococcus epidermidis biofilms on silicone catheters.

    PubMed

    Maisetta, Giuseppantonio; Grassi, Lucia; Di Luca, Mariagrazia; Bombardelli, Silvia; Medici, Chiara; Brancatisano, Franca Lisa; Esin, Semih; Batoni, Giovanna

    2016-08-01

    In search of new antimicrobials with anti-biofilm potential, in the present study activity of the frog-skin derived antimicrobial peptide temporin 1Tb (TB) against Staphylococcus epidermidis biofilms was investigated. A striking ability of TB to kill both forming and mature S. epidermidis biofilms was observed, especially when the peptide was combined with cysteine or EDTA, respectively. Kinetics studies demonstrated that the combination TB/EDTA was active against mature biofilms already after 2-4-h exposure. A double 4-h exposure of biofilms to TB/EDTA further increased the therapeutic potential of the same combination. Of note, TB/EDTA was able to eradicate S. epidermidis biofilms formed in vitro on silicone catheters. At eradicating concentrations, TB/EDTA did not cause hemolysis of human erythrocytes. The results shed light on the anti-biofilm properties of TB and suggest a possible application of the peptide in the lock therapy of catheters infected with S. epidermidis.

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

    PubMed

    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. Copyright © 2011 Elsevier Ltd. All rights reserved.

  18. Dual-species relations between Candida tropicalis isolated from apple juice ultrafiltration membranes, with Escherichia coli O157:H7 and Salmonella sp.

    PubMed

    Tarifa, M C; Lozano, J E; Brugnoni, L I

    2015-02-01

    The objective of this study was to determine the interactions between common spoilage yeast, Candida tropicalis, isolated from ultrafiltration membranes, and Escherichia coli O157:H7 and Salmonella sp. on stainless steel surfaces. Single and dual-species attachment assays were performed on stainless steel at 25°C using apple juice as culture medium. The growth of Salmonella sp. rose when it was co-cultivated with C. tropicalis in dual biofilms at 16 and 24 h; the same effect was observed for E. coli O157:H7 at 24 h. The colonization of C. tropicalis on stainless steel surfaces was reduced when it was co-cultivated with both pathogenic bacteria, reducing C. tropicalis population by at least 1.0 log unit. Visualization by SEM demonstrated that E. coli O157:H7 and Salmonella sp. adhere closely to hyphal elements using anchorage structures to attach to the surface and other cells. These results suggest a route for potential increased survival of pathogens in juice processing environments. These support the notion that the species involved interact in mixed yeast-bacteria communities favouring the development of bacteria over yeast. This study support the plausibility that pathogen interactions with strong biofilm forming members of spoilage microbiota, such as C. tropicalis, might play an important role for the survival and dissemination of E. coli O157:H7 and Salmonella sp. in food-processing environments. © 2014 The Society for Applied Microbiology.

  19. Phosphatidylcholine Membrane Fusion Is pH-Dependent.

    PubMed

    Akimov, Sergey A; Polynkin, Michael A; Jiménez-Munguía, Irene; Pavlov, Konstantin V; Batishchev, Oleg V

    2018-05-03

    Membrane fusion mediates multiple vital processes in cell life. Specialized proteins mediate the fusion process, and a substantial part of their energy is used for topological rearrangement of the membrane lipid matrix. Therefore, the elastic parameters of lipid bilayers are of crucial importance for fusion processes and for determination of the energy barriers that have to be crossed for the process to take place. In the case of fusion of enveloped viruses (e.g., influenza) with endosomal membrane, the interacting membranes are in an acidic environment, which can affect the membrane's mechanical properties. This factor is often neglected in the analysis of virus-induced membrane fusion. In the present work, we demonstrate that even for membranes composed of zwitterionic lipids, changes of the environmental pH in the physiologically relevant range of 4.0 to 7.5 can affect the rate of the membrane fusion notably. Using a continual model, we demonstrated that the key factor defining the height of the energy barrier is the spontaneous curvature of the lipid monolayer. Changes of this parameter are likely to be caused by rearrangements of the polar part of lipid molecules in response to changes of the pH of the aqueous solution bathing the membrane.

  20. Assessment of biofilm changes and concentration-depth profiles during arsenopyrite oxidation by Acidithiobacillus thiooxidans.

    PubMed

    Ramírez-Aldaba, Hugo; Vazquez-Arenas, Jorge; Sosa-Rodríguez, Fabiola S; Valdez-Pérez, Donato; Ruiz-Baca, Estela; García-Meza, Jessica Viridiana; Trejo-Córdova, Gabriel; Lara, René H

    2017-08-01

    Biofilm formation and evolution are key factors to consider to better understand the kinetics of arsenopyrite biooxidation. Chemical and surface analyses were carried out using Raman spectroscopy, scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), glow discharge spectroscopy (GDS), and protein analysis (i.e., quantification) in order to evaluate the formation of intermediate secondary compounds and any significant changes arising in the biofilm structure of Acidithiobacillus thiooxidans during a 120-h period of biooxidation. Results show that the biofilm first evolves from a low cell density structure (1 to 12 h) into a formation of microcolonies (24 to 120 h) and then finally becomes enclosed by a secondary compound matrix that includes pyrite (FeS 2 )-like, S n 2- /S 0 , and As 2 S 3 compounds, as shown by Raman and SEM-EDS. GDS analyses (concentration-depth profiles, i.e., 12 h) indicate significant differences for depth speciation between abiotic control and biooxidized surfaces, thus providing a quantitative assessment of surface-bulk changes across samples (i.e. reactivity and /or structure-activity relationship). Respectively, quantitative protein analyses and CLSM analyses suggest variations in the type of extracellular protein expressed and changes in the biofilm structure from hydrophilic (i.e., exopolysaccharides) to hydrophobic (i.e., lipids) due to arsenopyrite and cell interactions during the 120-h period of biooxidation. We suggest feasible environmental and industrial implications for arsenopyrite biooxidation based on the findings of this study.

  1. Membrane-based technologies for biogas separations.

    PubMed

    Basu, Subhankar; Khan, Asim L; Cano-Odena, Angels; Liu, Chunqing; Vankelecom, Ivo F J

    2010-02-01

    Over the past two decades, membrane processes have gained a lot of attention for the separation of gases. They have been found to be very suitable for wide scale applications owing to their reasonable cost, good selectivity and easily engineered modules. This critical review primarily focuses on the various aspects of membrane processes related to the separation of biogas, more in specific CO(2) and H(2)S removal from CH(4) and H(2) streams. Considering the limitations of inorganic materials for membranes, the present review will only focus on work done with polymeric materials. An overview on the performance of commercial membranes and lab-made membranes highlighting the problems associated with their applications will be given first. The development studies carried out to enhance the performance of membranes for gas separation will be discussed in the subsequent section. This review has been broadly divided into three sections (i) performance of commercial polymeric membranes (ii) performance of lab-made polymeric membranes and (iii) performance of mixed matrix membranes (MMMs) for gas separations. It will include structural modifications at polymer level, polymer blending, as well as synthesis of mixed matrix membranes, for which addition of silane-coupling agents and selection of suitable fillers will receive special attention. Apart from an overview of the different membrane materials, the study will also highlight the effects of different operating conditions that eventually decide the performance and longevity of membrane applications in gas separations. The discussion will be largely restricted to the studies carried out on polyimide (PI), cellulose acetate (CA), polysulfone (PSf) and polydimethyl siloxane (PDMS) membranes, as these membrane materials have been most widely used for commercial applications. Finally, the most important strategies that would ensure new commercial applications will be discussed (156 references).

  2. Hha Controls Escherichia coli O157:H7 Biofilm Formation by Differential Regulation of Global Transcriptional Regulators FlhDC and CsgD

    PubMed Central

    Bearson, Bradley L.

    2013-01-01

    Although molecular mechanisms promoting adherence of enterohemorrhagic Escherichia coli (EHEC) O157:H7 on epithelial cells are well characterized, regulatory mechanisms controlling biofilm formation are not fully understood. In this study, we demonstrate that biofilm formation in EHEC O157:H7 strain 86-24 is highly repressed compared to that in an isogenic hha mutant. The hha mutant produced large quantities of biofilm compared to the wild-type strain at 30°C and 37°C. Complementation of the hha mutant reduced the level of biofilm formation to that of the wild-type strain, indicating that Hha is a negative regulator of biofilm production. While swimming motility and expression of the flagellar gene fliC were significantly reduced, the expression of csgA (encoding curlin of curli fimbriae) and the ability to bind Congo red were significantly enhanced. The expression of both fliC and csgA and the phenotypes of motility and curli production affected by these two genes, respectively, were restored to wild-type levels in the complemented hha mutant. The csgA deletion abolished biofilm formation in the hha mutant and wild-type strain, and csgA complementation restored biofilm formation to these strains, indicating the importance of csgA and curli in biofilm formation. The regulatory effects of Hha on flagellar and curli gene expression appear to occur via the induction and repression of FlhDC and CsgD, as demonstrated by reduced flhD and increased csgD transcription in the hha mutant, respectively. In gel shift assays Hha interacted with flhDC and csgD promoters. In conclusion, Hha regulates biofilm formation in EHEC O157:H7 by differential regulation of FlhDC and CsgD, the global regulators of motility and curli production, respectively. PMID:23377937

  3. Hha controls Escherichia coli O157:H7 biofilm formation by differential regulation of global transcriptional regulators FlhDC and CsgD.

    PubMed

    Sharma, Vijay K; Bearson, Bradley L

    2013-04-01

    Although molecular mechanisms promoting adherence of enterohemorrhagic Escherichia coli (EHEC) O157:H7 on epithelial cells are well characterized, regulatory mechanisms controlling biofilm formation are not fully understood. In this study, we demonstrate that biofilm formation in EHEC O157:H7 strain 86-24 is highly repressed compared to that in an isogenic hha mutant. The hha mutant produced large quantities of biofilm compared to the wild-type strain at 30°C and 37°C. Complementation of the hha mutant reduced the level of biofilm formation to that of the wild-type strain, indicating that Hha is a negative regulator of biofilm production. While swimming motility and expression of the flagellar gene fliC were significantly reduced, the expression of csgA (encoding curlin of curli fimbriae) and the ability to bind Congo red were significantly enhanced. The expression of both fliC and csgA and the phenotypes of motility and curli production affected by these two genes, respectively, were restored to wild-type levels in the complemented hha mutant. The csgA deletion abolished biofilm formation in the hha mutant and wild-type strain, and csgA complementation restored biofilm formation to these strains, indicating the importance of csgA and curli in biofilm formation. The regulatory effects of Hha on flagellar and curli gene expression appear to occur via the induction and repression of FlhDC and CsgD, as demonstrated by reduced flhD and increased csgD transcription in the hha mutant, respectively. In gel shift assays Hha interacted with flhDC and csgD promoters. In conclusion, Hha regulates biofilm formation in EHEC O157:H7 by differential regulation of FlhDC and CsgD, the global regulators of motility and curli production, respectively.

  4. Inactivation of Escherichia coli O157:H7 in biofilm on food-contact surfaces by sequential treatments of aqueous chlorine dioxide and drying.

    PubMed

    Bang, Jihyun; Hong, Ayoung; Kim, Hoikyung; Beuchat, Larry R; Rhee, Min Suk; Kim, Younghoon; Ryu, Jee-Hoon

    2014-11-17

    We investigated the efficacy of sequential treatments of aqueous chlorine and chlorine dioxide and drying in killing Escherichia coli O157:H7 in biofilms formed on stainless steel, glass, plastic, and wooden surfaces. Cells attached to and formed a biofilm on wooden surfaces at significantly (P ≤ 0.05) higher levels compared with other surface types. The lethal activities of sodium hypochlorite (NaOCl) and aqueous chlorine dioxide (ClO₂) against E. coli O157:H7 in a biofilm on various food-contact surfaces were compared. Chlorine dioxide generally showed greater lethal activity than NaOCl against E. coli O157:H7 in a biofilm on the same type of surface. The resistance of E. coli O157:H7 to both sanitizers increased in the order of wood>plastic>glass>stainless steel. The synergistic lethal effects of sequential ClO₂ and drying treatments on E. coli O157:H7 in a biofilm on wooden surfaces were evaluated. When wooden surfaces harboring E. coli O157:H7 biofilm were treated with ClO₂ (200 μg/ml, 10 min), rinsed with water, and subsequently dried at 43% relative humidity and 22 °C, the number of E. coli O157:H7 on the surface decreased by an additional 6.4 CFU/coupon within 6 h of drying. However, when the wooden surface was treated with water or NaOCl and dried under the same conditions, the pathogen decreased by only 0.4 or 1.0 log CFU/coupon, respectively, after 12 h of drying. This indicates that ClO₂ treatment of food-contact surfaces results in residual lethality to E. coli O157:H7 during the drying process. These observations will be useful when selecting an appropriate type of food-contact surfaces, determining a proper sanitizer for decontamination, and designing an effective sanitization program to eliminate E. coli O157:H7 on food-contact surfaces in food processing, distribution, and preparation environments. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Acceleration of pH variation in cloudy apple juice using electrodialysis with bipolar membranes.

    PubMed

    Lam Quoc, A; Lamarche, F; Makhlouf, J

    2000-06-01

    The purpose of this study was to accelerate pH variation in cloudy apple juice using electrodialysis (ED). The testing was conducted using two ED configurations. The bipolar and cationic membrane configuration showed that reducing the spacing from 8 to 0.75 mm had little effect on treatment time, whereas stacking eight bipolar membranes reduced acidification time by 30%, although the treatment still took too long (21 min). Furthermore, it was not possible to acidify apple juice to a pH of 2.0 to completely inhibit enzymatic browning. The bipolar and anionic membrane configuration helped to accelerate the acidification step by a factor of 3, increasing the yield from 3.3 to 10 L of juice/m(2) membrane/min. Moreover, treatment time was inversely proportional to the size of the membrane stack. The speed at which the pH of acidified juice returned to its initial value was, however, 4 times slower than the speed of acidification, giving a yield of 2.5 L of juice/m(2) membrane/min. By accelerating the acidification step, ED treatment with bipolar and anionic membranes results in more effective polyphenol oxidase activity and more rapid control of juice browning at pH 2.0. Also, the treatment has very little effect on the chemical composition and organoleptic quality of apple juice.

  6. Acoustic vibration can enhance bacterial biofilm formation.

    PubMed

    Murphy, Mark F; Edwards, Thomas; Hobbs, Glyn; Shepherd, Joanna; Bezombes, Frederic

    2016-12-01

    This paper explores the use of low-frequency-low-amplitude acoustic vibration on biofilm formation. Biofilm development is thought to be governed by a diverse range of environmental signals and much effort has gone into researching the effects of environmental factors including; nutrient availability, pH and temperature on the growth of biofilms. Many biofilm-forming organisms have evolved to thrive in mechanically challenging environments, for example soil yet, the effects of the physical environment on biofilm formation has been largely ignored. Exposure of Pseudomonas aeruginosa to vibration at 100, 800 and 1600 Hz for 48 h, resulted in a significant increase in biofilm formation compared with the control, with the greatest growth seen at 800 Hz vibration. The results also show that this increase in biofilm formation is accompanied with an increase in P. aeruginosa cell number. Acoustic vibration was also found to regulate the spatial distribution of biofilm formation in a frequency-dependent manner. Exposure of Staphylococcus aureus to acoustic vibration also resulted in enhanced biofilm formation with the greatest level of biofilm being formed following 48 h exposure at 1600 Hz. These results show that acoustic vibration can be used to control biofilm formation and therefore presents a novel and potentially cost effective means to manipulate the development and yield of biofilms in a range of important industrial and medical processes. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  7. Inhibitory activity of isoniazid and ethionamide against Cryptococcus biofilms.

    PubMed

    Cordeiro, Rossana de Aguiar; Serpa, Rosana; Marques, Francisca Jakelyne de Farias; de Melo, Charlline Vládia Silva; Evangelista, Antonio José de Jesus; Mota, Valquíria Ferreira; Brilhante, Raimunda Sâmia Nogueira; Bandeira, Tereza de Jesus Pinheiro Gomes; Rocha, Marcos Fábio Gadelha; Sidrim, José Júlio Costa

    2015-11-01

    In recent years, the search for drugs to treat systemic and opportunistic mycoses has attracted great interest from the scientific community. This study evaluated the in vitro inhibitory effect of the antituberculosis drugs isoniazid and ethionamide alone and combined with itraconazole and fluconazole against biofilms of Cryptococcus neoformans and Cryptococcus gattii. Antimicrobials were tested at defined concentrations after susceptibility assays with Cryptococcus planktonic cells. In addition, we investigated the synergistic interaction of antituberculosis drugs and azole derivatives against Cryptococcus planktonic cells, as well as the influence of isoniazid and ethionamide on ergosterol content and cell membrane permeability. Isoniazid and ethionamide inhibited both biofilm formation and viability of mature biofilms. Combinations formed by antituberculosis drugs and azoles proved synergic against both planktonic and sessile cells, showing an ability to reduce Cryptococcus biofilms by approximately 50%. Furthermore, isoniazid and ethionamide reduced the content of ergosterol in Cryptococcus spp. planktonic cells and destabilized or permeabilized the fungal cell membrane, leading to leakage of macromolecules. Owing to the paucity of drugs able to inhibit Cryptococcus biofilms, we believe that the results presented here might be of interest in the designing of new antifungal compounds.

  8. Expression of a constitutively activated plasma membrane H+-ATPase in Nicotiana tabacum BY-2 cells results in cell expansion.

    PubMed

    Niczyj, Marta; Champagne, Antoine; Alam, Iftekhar; Nader, Joseph; Boutry, Marc

    2016-11-01

    Increased acidification of the external medium by an activated H + -ATPase results in cell expansion, in the absence of upstream activating signaling. The plasma membrane H + -ATPase couples ATP hydrolysis with proton transport outside the cell, and thus creates an electrochemical gradient, which energizes secondary transporters. According to the acid growth theory, this enzyme is also proposed to play a major role in cell expansion, by acidifying the external medium and so activating enzymes that are involved in cell wall-loosening. However, this theory is still debated. To challenge it, we made use of a plasma membrane H + -ATPase isoform from Nicotiana plumbaginifolia truncated from its C-terminal auto-inhibitory domain (ΔCPMA4), and thus constitutively activated. This protein was expressed in Nicotiana tabacum BY-2 suspension cells using a heat shock inducible promoter. The characterization of several independent transgenic lines showed that the expression of activated ΔCPMA4 resulted in a reduced external pH by 0.3-1.2 units, as well as in an increased H + -ATPase activity by 77-155 % (ATP hydrolysis), or 70-306 % (proton pumping) of isolated plasma membranes. In addition, ΔCPMA4-expressing cells were 17-57 % larger than the wild-type cells and displayed abnormal shapes. A proteomic comparison of plasma membranes isolated from ΔCPMA4-expressing and wild-type cells revealed the altered abundance of several proteins involved in cell wall synthesis, transport, and signal transduction. In conclusion, the data obtained in this work showed that H + -ATPase activation is sufficient to induce cell expansion and identified possible actors which intervene in this process.

  9. Antimicrobial peptide AMPNT-6 from Bacillus subtilis inhibits biofilm formation by Shewanella putrefaciens and disrupts its preformed biofilms on both abiotic and shrimp shell surfaces.

    PubMed

    Deng, Qi; Pu, Yuehua; Sun, Lijun; Wang, Yaling; Liu, Yang; Wang, Rundong; Liao, Jianmeng; Xu, Defeng; Liu, Ying; Ye, Riying; Fang, Zhijia; Gooneratne, Ravi

    2017-12-01

    Shewanella putrefaciens biofilm formation is of great concern for the shrimp industry because it adheres easily to food and food-contact surfaces and is a source of persistent and unseen contamination that causes shrimp spoilage and economic losses to the shrimp industry. Different concentrations of an antimicrobial lipopeptide, the fermentation product of Bacillus subtilis, AMPNT-6, were tested for the ability to reduce adhesion and disrupt S. putrefaciens preformed biofilms on two different contact surfaces (shrimp shell, stainless steel sheet). AMPNT-6 displayed a marked dose- and time-dependent anti-adhesive effect>biofilm removal. 3MIC AMPNT-6 was able both to remove biofilm and prevent bacteria from forming biofilm in a 96-well polystyrene microplate used as the model surface. 2MIC AMPNT-6 prevented bacteria from adhering to the microplate surface to form biofilm for 3h and removed already existing biofilm within 24h. Secretion of extracellular polymeric substances incubated in LB broth for 24h by S. putrefaciens was minimal at 3× MIC AMPNT-6. Scanning electron microscopy showed that damage to S. putrefaciens bacteria by AMPNT-6 possibly contributed to the non-adherence to the surfaces. Disruption of the mature biofilm structure by AMPNT-6 contributed to biofilm removal. It is concluded that AMPNT-6 can be used effectively to prevent attachment and also detach S. putrefaciens biofilms from shrimp shells, stainless steel sheets and polystyrene surfaces. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Precise detection of pH inside large unilamellar vesicles using membrane-impermeable dendritic porphyrin-based nanoprobes.

    PubMed

    Leiding, Thom; Górecki, Kamil; Kjellman, Tomas; Vinogradov, Sergei A; Hägerhäll, Cecilia; Arsköld, Sindra Peterson

    2009-05-15

    Accurate real-time measurements of proton concentration gradients are pivotal to mechanistic studies of proton translocation by membrane-bound enzymes. Here we report a detailed characterization of the pH-sensitive fluorescent nanoprobe Glu(3), which is well suited for pH measurements in microcompartmentalized biological systems. The probe is a polyglutamic porphyrin dendrimer in which multiple carboxylate termini ensure its high water solubility and prevent its diffusion across phospholipid membranes. The probe's pK is in the physiological pH range, and its protonation can be followed ratiometrically by absorbance or fluorescence in the ultraviolet-visible spectral region. The usefulness of the probe was enhanced by using a semiautomatic titration system coupled to a charge-coupled device (CCD) spectrometer, enabling fast and accurate titrations and full spectral coverage of the system at millisecond time resolution. The probe's pK was measured in bulk solutions as well as inside large unilamellar vesicles in the presence of physiologically relevant ions. Glu(3) was found to be completely membrane impermeable, and its distinct spectroscopic features permit pH measurements inside closed membrane vesicles, enabling quantitative mechanistic studies of membrane-spanning proteins. Performance of the probe was demonstrated by monitoring the rate of proton leakage through the phospholipid bilayer in large vesicles with and without the uncoupler gramicidin present. Overall, as a probe for biological proton translocation measurements, Glu(3) was found to be superior to the commercially available pH indicators.

  11. Candida/Candida biofilms. First description of dual-species Candida albicans/C. rugosa biofilm.

    PubMed

    Martins, Carlos Henrique Gomes; Pires, Regina Helena; Cunha, Aline Oliveira; Pereira, Cristiane Aparecida Martins; Singulani, Junya de Lacorte; Abrão, Fariza; Moraes, Thais de; Mendes-Giannini, Maria José Soares

    2016-04-01

    Denture liners have physical properties that favour plaque accumulation and colonization by Candida species, irritating oral tissues and causing denture stomatitis. To isolate and determine the incidence of oral Candida species in dental prostheses, oral swabs were collected from the dental prostheses of 66 patients. All the strains were screened for their ability to form biofilms; both monospecies and dual-species combinations were tested. Candida albicans (63 %) was the most frequently isolated microorganism; Candida tropicalis (14 %), Candida glabrata (13 %), Candida rugosa (5 %), Candida parapsilosis (3 %), and Candida krusei (2 %) were also detected. The XTT assay showed that C. albicans SC5314 possessed a biofilm-forming ability significantly higher (p < 0.001) than non-albicans Candida strains, after 6 h 37 °C. The total C. albicans CFU from a dual-species biofilm was less than the total CFU of a monospecies C. albicans biofilm. In contrast to the profuse hyphae verified in monospecies C. albicans biofilms, micrographies showed that the C. albicans/non-albicans Candida biofilms consisted of sparse yeast forms and profuse budding yeast cells that generated a network. These results suggested that C. albicans and the tested Candida species could co-exist in biofilms displaying apparent antagonism. The study provide the first description of C. albicans/C. rugosa mixed biofilm. Copyright © 2016 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  12. Prospective role of indigenous Exiguobacterium profundum PT2 in arsenic biotransformation and biosorption by planktonic cultures and biofilms.

    PubMed

    Saba; Andreasen, R; Li, Y; Rehman, Y; Ahmed, M; Meyer, R L; Sabri, A N

    2018-02-01

    The aim of this study was to analyse arsenic (As) transformation and biosorption by indigenous As-resistant bacteria both in planktonic and biofilm modes of growth. As-resistant bacteria were isolated from industrial waste water and strain PT2, and identified as Exiguobacterium profundum through 16S rRNA gene sequencing was selected for further study. As transformation and biosorption by E. profundumPT2 was determined by HPLC-ICP-MS analysis. Planktonic cultures reduced 3·73 mmol l -1 As 5+ into As 3+ from artificial waste water effluent after 48-h incubation. In case of biosorption, planktonic cultures and biofilms exhibited 25·2 and 29·4 mg g -1 biomass biosorption, respectively. As biosorption kinetics followed Freundlich isotherm and pseudo second-order model. Biofilm formation peaked after 3 days of incubation, and in the presence of As stress, biofilm formation was significantly affected in contrast to control (P < 0·05). Homogeneous nature of mature biofilms with an increased demand of nutrients was revealed by minimum roughness and maximum surface to biovolume ratio measured through CLSM analysis. Indigenous As-resistant E. profundumPT2 was found capable of As transformation and biosorption both in the form of planktonic cultures and biofilms. Indigenous biofilm forming E. profundum PT2 revealing As biosorption and biotransformation potential is presented an eco-friendly and cost-effective source for As remediation that can be implemented for waste water treatment. © 2017 The Society for Applied Microbiology.

  13. Photo Inactivation of Streptococcus mutans Biofilm by Violet-Blue light.

    PubMed

    Gomez, Grace F; Huang, Ruijie; MacPherson, Meoghan; Ferreira Zandona, Andrea G; Gregory, Richard L

    2016-09-01

    Among various preventive approaches, non-invasive phototherapy/photodynamic therapy is one of the methods used to control oral biofilm. Studies indicate that light at specific wavelengths has a potent antibacterial effect. The objective of this study was to determine the effectiveness of violet-blue light at 380-440 nm to inhibit biofilm formation of Streptococcus mutans or kill S. mutans. S. mutans UA159 biofilm cells were grown for 12-16 h in 96-well flat-bottom microtiter plates using tryptic soy broth (TSB) or TSB with 1 % sucrose (TSBS). Biofilm was irradiated with violet-blue light for 5 min. After exposure, plates were re-incubated at 37 °C for either 2 or 6 h to allow the bacteria to recover. A crystal violet biofilm assay was used to determine relative densities of the biofilm cells grown in TSB, but not in TSBS, exposed to violet-blue light. The results indicated a statistically significant (P < 0.05) decrease compared to the non-treated groups after the 2 or 6 h recovery period. Growth rates of planktonic and biofilm cells indicated a significant reduction in the growth rate of the violet-blue light-treated groups grown in TSB and TSBS. Biofilm viability assays confirmed a statistically significant difference between violet-blue light-treated and non-treated groups in TSB and TSBS. Visible violet-blue light of the electromagnetic spectrum has the ability to inhibit S. mutans growth and reduce the formation of S. mutans biofilm. This in vitro study demonstrated that violet-blue light has the capacity to inhibit S. mutans biofilm formation. Potential clinical applications of light therapy in the future remain bright in preventing the development and progression of dental caries.

  14. A simple 2D biofilm model yields a variety of morphological features.

    PubMed

    Hermanowicz, S W

    2001-01-01

    A two-dimensional biofilm model was developed based on the concept of cellular automata. Three simple, generic processes were included in the model: cell growth, internal and external mass transport and cell detachment (erosion). The model generated a diverse range of biofilm morphologies (from dense layers to open, mushroom-like forms) similar to those observed in real biofilm systems. Bulk nutrient concentration and external mass transfer resistance had a large influence on the biofilm structure.

  15. Identification of Biofilm Matrix-Associated Proteins from an Acid Mine Drainage Microbial Community

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jiao, Yongqin; D'Haeseleer, Patrik M; Dill, Brian

    2011-01-01

    In microbial communities, extracellular polymeric substances (EPS), also called the extracellular matrix, provide the spatial organization and structural stability during biofilm development. One of the major components of EPS is protein, but it is not clear what specific functions these proteins contribute to the extracellular matrix or to microbial physiology. To investigate this in biofilms from an extremely acidic environment, we used shotgun proteomics analyses to identify proteins associated with EPS in biofilms at two developmental stages, designated DS1 and DS2. The proteome composition of the EPS was significantly different from that of the cell fraction, with more than 80%more » of the cellular proteins underrepresented or undetectable in EPS. In contrast, predicted periplasmic, outer membrane, and extracellular proteins were overrepresented by 3- to 7-fold in EPS. Also, EPS proteins were more basic by 2 pH units on average and about half the length. When categorized by predicted function, proteins involved in motility, defense, cell envelope, and unknown functions were enriched in EPS. Chaperones, such as histone-like DNA binding protein and cold shock protein, were overrepresented in EPS. Enzymes, such as protein peptidases, disulfide-isomerases, and those associated with cell wall and polysaccharide metabolism, were also detected. Two of these enzymes, identified as -N-acetylhexosaminidase and cellulase, were confirmed in the EPS fraction by enzymatic activity assays. Compared to the differences between EPS and cellular fractions, the relative differences in the EPS proteomes between DS1 and DS2 were smaller and consistent with expected physiological changes during biofilm development.« less

  16. The Fusobacterium nucleatum Outer Membrane Protein RadD Is an Arginine-Inhibitable Adhesin Required for Inter-Species Adherence and the Structured Architecture of Multi-Species Biofilm

    PubMed Central

    Kaplan, Christopher W.; Lux, Renate; Haake, Susan Kinder; Shi, Wenyuan

    2009-01-01

    Summary A defining characteristic of the suspected periodontal pathogen Fusobacterium nucleatum is its ability to adhere to a plethora of oral bacteria. This distinguishing feature is suggested to play an important role in oral biofilm formation and pathogenesis, with fusobacteria proposed to serve as central “bridging organisms” in the architecture of the oral biofilm bringing together species which would not interact otherwise. Previous studies indicate that these bacterial interactions are mediated by galactose- or arginine-inhibitable adhesins although genetic evidence for the role and nature of these proposed adhesins remains elusive. To characterize these adhesins at the molecular level, the genetically transformable F. nucleatum strain ATCC 23726 was screened for adherence properties, and arginine inhibitable adhesion was evident, while galactose-inhibitable adhesion was not detected. Six potential arginine binding proteins were isolated from the membrane fraction of F. nucleatum ATCC 23726 and identified via mass spectroscopy as members of the outer membrane family of proteins in F. nucleatum. Inactivation of the genes encoding these six candidates for arginine-inhibitable adhesion and two additional homologues revealed that only a mutant derivative carrying an insertion in Fn1526 (now designated as radD) demonstrated significantly decreased co-aggregation with representatives of the Gram-positive “early oral colonizers”. Lack of the 350 kDa outer membrane protein encoded by radD resulted in the failure to form the extensive structured biofilm observed with the parent strain when grown in the presence of Streptococcus sanguinis ATCC 10556. These findings indicate that radD is responsible for arginine-inhibitable adherence of F. nucleatum and provides definitive molecular evidence that F. nucleatum adhesins play a vital role in inter-species adherence and multispecies biofilm formation. PMID:19007407

  17. Physically Gelled Room-Temperature Ionic Liquid-Based Composite Membranes for CO2/N-2 Separation: Effect of Composition and Thickness on Membrane Properties and Performance

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nguyen, PT; Voss, BA; Wiesenauer, EF

    2013-07-03

    An aspartame-based, low molecular-weight organic gelator (LMOG) was used to form melt-infused and composite membranes with two different imidazolium-based room-temperature ionic liquids (RTILs) for CO2 separation from N-2. Previous work demonstrated that LMOGs can gel RTILs at low, loading levels, and this aspartame-based LMOG was selected because it has been reported to gel a large number of RTILs. The imidazolium-based RTILs were used because of their inherent good properties for CO2/light gas separations. Analysis of the resulting bulk RTIL/LMOG physical gels showed that these materials have high sol-gel transition temperatures (ca. 135 degrees C) suitable for flue gas applications. Gasmore » permeabilities and burst pressure measurements of thick, melt infused membranes revealed a trade-off between high CO2 permeabilities and good mechanical stability as a function of the LMOG loading. Defect-free, composite membranes of the gelled RTILs were successfully fabricated by choosing an appropriate porous membrane support (hydrophobic PTFE) using a suitable coating technique (roller coating). The thicknesses of the applied composite gel layers ranged from 10.3 to 20.7 mu m, which represents an order of magnitude decrease in active layer thickness, compared to the original melt-infused gel RTIL membranes.« less

  18. Structurally stable graphene oxide-based nanofiltration membranes with bioadhesive polydopamine coating

    NASA Astrophysics Data System (ADS)

    Wang, Chongbin; Li, Zhiyuan; Chen, Jianxin; Yin, Yongheng; Wu, Hong

    2018-01-01

    Graphene oxide (GO)-based membranes possess promising potential in liquid separation for its high flux. The state-of-art GO-based membranes need to be supported by a substrate to ensure that the ultra-thin GO layer can withstand transmembrane pressure in practical applications. The interfacial compatibility of this kind of composite membrane remains a great challenge due to the intrinsic difference in chemical/physical properties between the GO sheets and the substrate. In this paper, a structurally stable GO-based composite nanofiltration membrane was fabricated by coupling the mussel-inspired adhesive platform and filtration-assisted assembly of GO laminates. The water flux for the prepared GO-based nanofiltration membrane reached up to 85 L m-2 h-1 bar-1 with a high retention above 95% and 100% for Orange G and Congo Red, respectively. The membrane exhibited highly stable structure owing to the covalent and noncovalent interactions between GO separation layer and dopamine adhesive platform.

  19. Colonization dynamics of biofilm-associated ciliate morphotypes at different flow velocities.

    PubMed

    Risse-Buhl, Ute; Küsel, Kirsten

    2009-01-01

    The impact of flow velocity on initial ciliate colonization dynamics on surfaces were studied in the third order Ilm stream (Thuringia, Germany) at a slow flowing site (0.09ms(-1)) and two faster flowing sites (0.31ms(-1)) and in flow channels at 0.05, 0.4, and 0.8ms(-1). At the slow flowing stream site, surfaces were rapidly colonized by ciliates with up to 60 cells cm(-2) after 24h. In flow channels, the majority of suspended ciliates and inorganic matter accumulated at the surface within 4.5h at 0.05ms(-1). At 0.4ms(-1) the increase in ciliate abundance in the biofilm was highest between 72 and 168h at about 3 cells cm(-2)h(-1). Faster flow velocities were tolerated by vagile flattened ciliates that live in close contact to the surface. Vagile flattened and round filter feeders preferred biofilms at slow flow velocities. Addition of inorganic particles (0, 0.6, and 7.3mgcm(-2)) did not affect ciliate abundance in flow channel biofilms, but small ciliate species dominated and number of species was lowest (16 species cm(-2)) in biofilms at high sediment content. Although different morphotypes dominated the communities at contrasting flow velocities, all functional groups contributed to initial biofilm communities implementing all trophic links within the microbial loop.

  20. Low concentrations of Al(III) accelerate the formation of biofilm: Multiple effects of hormesis and flocculation.

    PubMed

    Cui, Xiaochun; Huo, Mingxin; Chen, Congli; Yu, Zhisen; Zhou, Chen; Li, Anran; Qiao, Bingqian; Zhou, Dandan; Crittenden, John C

    2018-09-01

    Residual Al(III) (at low concentration) is common in water treatment plants (WTPs) and is associated with bacteria. We hypothesize that Al(III) accelerate biofouling due to its hydrolysis and hormesis characteristics, as compared with other cations. To verify this, we elaborated the roles of Al(III) at low concentrations on the biofilm formation. Al(III) hormesis (<2.0mg/L) stimulated bacteria growth increased by ~3.7 times, and extracellular polymeric substances production also enhanced. Al(III) flocculation resulted in the suspended cells precipitation instantly, for Al(III) dosages of 0.6 and 2.0mg/L and the concentration of Al(III) decreased by 0.07 and 0.14mg/L, respectively. Al(III) poisoned the bridged bacterial cells and decreased their ATP by 22.36% and 55.91%, respectively. Al(III) formed polymer presented strong affinity with bacterial outer membrane, and this damaged the bacterial outer membrane. This caused proteins to leak at the combined point. Al-polymer bound to NH 2 and/or NH on the leaked protein, contributed to biofilm formation. Biofilm maturity was aided by polysaccharides, which shielded Al(III) toxicity for the formed biofilm. Thus, the biofilm exhibited a distinguished double-layer microstructure, principally with proteins and inactivated cells at the bottom, polysaccharides and activated cells at the top. Thus, hormesis and flocculation caused by low concentration Al(III) mutually promoted each other, and together accelerated biofilm formation. Copyright © 2018 Elsevier B.V. All rights reserved.

  1. Turbulence accelerates the growth of drinking water biofilms.

    PubMed

    Tsagkari, E; Sloan, W T

    2018-06-01

    Biofilms are found at the inner surfaces of drinking water pipes and, therefore, it is essential to understand biofilm processes to control their formation. Hydrodynamics play a crucial role in shaping biofilms. Thus, knowing how biofilms form, develop and disperse under different flow conditions is critical in the successful management of these systems. Here, the development of biofilms after 4 weeks, the initial formation of biofilms within 10 h and finally, the response of already established biofilms within 24-h intervals in which the flow regime was changed, were studied using a rotating annular reactor under three different flow regimes: turbulent, transition and laminar. Using fluorescence microscopy, information about the number of microcolonies on the reactor slides, the surface area of biofilms and of extracellular polymeric substances and the biofilm structures was acquired. Gravimetric measurements were conducted to characterise the thickness and density of biofilms, and spatial statistics were used to characterise the heterogeneity and spatial correlation of biofilm structures. Contrary to the prevailing view, it was shown that turbulent flow did not correlate with a reduction in biofilms; turbulence was found to enhance both the initial formation and the development of biofilms on the accessible surfaces. Additionally, after 24-h changes of the flow regime it was indicated that biofilms responded to the quick changes of the flow regime. Overall, this work suggests that different flow conditions can cause substantial changes in biofilm morphology and growth and specifically that turbulent flow can accelerate biofilm growth in drinking water.

  2. Contributions of EspA Filaments and Curli Fimbriae in Cellular Adherence and Biofilm Formation of Enterohemorrhagic Escherichia coli O157:H7

    PubMed Central

    Sharma, Vijay K.; Kudva, Indira T.; Bearson, Bradley L.; Stasko, Judith A.

    2016-01-01

    In Escherichia coli O157:H7 (O157), the filamentous structure of the type III secretion system is produced from the polymerization of the EspA protein. EspA filaments are essential for O157 adherence to epithelial cells. In previous studies, we demonstrated that O157 hha deletion mutants showed increased adherence to HEp-2 cells and produced abundant biofilms. Transcriptional analysis revealed increased expression of espA as well as the csgA gene, which encodes curli fimbriae that are essential for biofilm formation. In the present study, we constructed hha espA, hha csgA, and hha csgA espA deletion mutants to determine the relative importance of EspA and CsgA in O157 adherence to HEp-2 cells and biofilm formation. In vitro adherence assays, conducted at 37°C in a tissue culture medium containing 0.1% glucose, showed that HEp-2 cell adherence required EspA because hha espA and hha csgA espA mutants adhered to HEp-2 cells at higher levels only when complemented with an espA-expressing plasmid. Biofilm assays performed at 28°C in a medium lacking glucose showed dependency of biofilm formation on CsgA; however EspA was not produced under these conditions. Despite production of detectable levels of EspA at 37°C in media supplemented with 0.1% glucose, the biofilm formation occurred independent of EspA. These results indicate dependency of O157 adherence to epithelial cells on EspA filaments, while CsgA promoted biofilm formation under conditions mimicking those found in the environment (low temperature with nutrient limitations) and in the digestive tract of an host animal (higher temperature and low levels of glucose). PMID:26900701

  3. Effect of biofilm dryness on the transfer of Listeria monocytogenes biofilms grown on stainless steel to bologna and hard salami.

    PubMed

    Rodríguez, Andrés; Autio, Wesley R; McLandsborough, Lynne A

    2007-11-01

    Listeria monocytogenes continues to be a major cause of class I food recalls in the United States. Very little is known about its transfer and cross-contamination in processing scenarios. The objective of this study was to evaluate the effect of hydration level on L. monocytogenes biofilms grown on stainless steel and its effect on the biofilm transfer to foods. Biofilms were grown on stainless steel in diluted tryptic soy broth 1:20 for 48 h at 32 degrees C. After this, biofilms were equilibrated over saturated salt solutions at 20 degrees C for 24 h (94, 75, 58, and 33% relative humidity; % RH) prior to transferring. Transfer experiments were conducted from inoculated stainless steel to bologna and hard salami at a constant pressure (45 kPa) and time (30 s) with a universal testing machine. The experiment was designed with a factorial design 4 x 2 (biofilms equilibrated at 4% RH and two foods) and duplicated every day, and the whole experiment was repeated nine times. The results were analyzed with an analysis of variance by SAS Statistical Analysis Software. Our results showed that more bacteria were transferred to bologna (mean efficiency of transfer [EOT] = 3.0) than to hard salami (mean EOT = 0.35, P < 0.01). As biofilms became drier, the transfer of Listeria from stainless steel to both foods increased (P < 0.05). The EOT increased from 2 to 3.8 and from 0.2 to 0.51 upon transfer when drying the biofilm for bologna and hard salami, respectively. This study may be an indication that as biofilms were dried, the cell-cell and cell-surface interactions became weaker, and bacterial transfer increased. This phenomenon was enhanced in foods containing higher water activity levels. We hypothesize that this increased in transfer was due to the presence of capillary forces in the food.

  4. Photodynamic therapy for inactivating endodontic bacterial biofilms and effect of tissue inhibitors on antibacterial efficacy

    NASA Astrophysics Data System (ADS)

    Shrestha, Annie; Kishen, Anil

    Complex nature of bacterial cell membrane and structure of biofilm has challenged the efficacy of antimicrobial photodynamic therapy (APDT) to achieve effective disinfection of infected root canals. In addition, tissue-inhibitors present inside the root canals are known to affect APDT activity. This study was aimed to assess the effect of APDT on bacterial biofilms and evaluate the effect of tissue-inhibitors on the APDT. Rose-bengal (RB) and methylene-blue (MB) were tested on Enterococcus faecalis (gram-positive) and Pseudomonas aeruginosa (gram-negative) biofilms. In vitro 7- day old biofilms were sensitized with RB and MB, and photodynamically activated with 20-60 J/cm2. Photosensitizers were pre-treated with different tissue-inhibitors (dentin, dentin-matrix, pulp tissue, bacterial lipopolysaccharides (LPS), and bovine serum albumin (BSA)) and tested for antibacterial effect of APDT. Microbiological culture based analysis was used to analyze the cell viability, while Laser Scanning Confocal Microscopy (LSCM) was used to examine the structure of biofilm. Photoactivation resulted in significant reduction of bacterial biofilms with RB and MB. The structure of biofilm under LSCM was found to be disrupted with reduced biofilm thickness. Complete biofilm elimination could not be achieved with both tested photosensitizers. APDT effect using MB and RB was inhibited in a decreasing order by dentin-matrix, BSA, pulp, dentin and LPS (P< 0.05). Both strains of bacterial biofilms resisted complete elimination after APDT and the tissue inhibitors existing within the root canal reduced the antibacterial activity at varying degrees. Further research is required to enhance the antibacterial efficacy of APDT in an endodontic environment.

  5. Dynamics of Nafion membrane swelling in H2O/D2O mixtures as studied using FTIR technique

    NASA Astrophysics Data System (ADS)

    Bunkin, Nikolai F.; Kozlov, Valeriy A.; Shkirin, Alexey V.; Ninham, Barry W.; Balashov, Anatoliy A.; Gudkov, Sergey V.

    2018-03-01

    Experiments with Fourier transform spectrometry of Nafion, a water-swollen polymeric membrane, are described. The transmittance spectra of liquid samples and Nafion, soaked in these samples, were studied, depending on the deuterium content in water in the spectral range 1.8-2.15 μm. The experiments were carried out using two protocols: in the first protocol we studied the dynamics of Nafion swelling in H2O + D2O mixtures for the deuterium concentrations 3 < C < 104 ppm, and in the second protocol we studied the dynamics of swelling in pure heavy water (C = 106 ppm). For liquid mixtures in the concentration range 3 < C < 104 ppm, the transmittance spectra are the same, but for Nafion soaked in these fluids, the corresponding spectra are different. It is shown that, in the range of deuterium contents C = 90-500 ppm, the behavior of transmittance of the polymer membrane is non-monotonic. In experiments using the second protocol, the dynamics of diffusion replacement of residual water, which is always present in the bulk of the polymer membrane inside closed cavities (i.e., without access to atmospheric air), were studied. The experimentally estimated diffusion coefficient for this process is ≈6.10-11 cm2/s.

  6. In Vitro Study of Bacteriophage AB3 Endolysin LysAB3 Activity Against Acinetobacter baumannii Biofilm and Biofilm-Bound A. baumannii.

    PubMed

    Zhang, Jie; Xu, Lu-Lu; Gan, Dan; Zhang, Xingping

    2018-06-01

    The increase in the prevalence of drug-resistant Acinetobacter baumannii is a serious public health concern, which is closely linked to the formation of biofilm. It is reported that the bacteriophage and its endolysin have a good ability to degrade biofilms. The goals of this study were to compare the ability of A. baumannii bacteriophage AB3, its endolysin AB3, and three antibiotics to degrade A. baumannii biofilm and biofilm-bound A. baumannii and to understand the antibacterial mechanism of LysAB3. The 558-bp sequence of the LysAB3 gene was amplified by polymerase chain reaction (PCR); the fragment was cloned into pET28a (+) to construct the recombinant plasmid pET28a-LysAB3, which was then expressed in E. coli BL21 (DE3) to obtain the LysAB3. Differences in A. baumannii biofilm and biofilm-bound A. baumannii after treatment with bacteriophage AB3, LysAB3 or three antibiotics were examined using the crystal violet staining method and an MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide) assay. Changes in biofilm morphology and thickness in each treatment group were observed by laser scanning confocal microscopy. In addition, a LysAB3 construct with the amphiphilic peptide structural region removed (LysAB3-D) was assessed for its antibacterial activity. After 24-hour treatment with either bacteriophage AB3 and its LysAB3, A. baumannii biofilms were significantly degraded, and the number of viable biofilm-bound A. baumannii were also significantly decreased. After removing the amphiphilic peptide structure motif from LysAB3, the antibacterial activity decreased from 95.8% to 33.3%. Thus, LysAB3 can effectively degrade A. baumannii biofilm and biofilm-bound A. baumannii in vitro. The antibacterial mechanism of LysAB3 may be associated with the ability of the amphiphilic peptide structural region to enhance the permeability of cytoplasmic membrane of A. baumannii by degradation of bacterial wall peptidoglycan.

  7. Efficacy of NVC-422 against Staphylococcus aureus biofilms in a sheep biofilm model of sinusitis.

    PubMed

    Singhal, Deepti; Jekle, Andreas; Debabov, Dmitri; Wang, Lu; Khosrovi, Bez; Anderson, Mark; Foreman, Andrew; Wormald, Peter-John

    2012-01-01

    Bacterial biofilms are a major obstacle in management of recalcitrant chronic rhinosinusitis. NVC-422 is a potent, fast-acting, broad-spectrum, nonantibiotic, antimicrobial with a new mechanism of action effective against biofilm bacteria in in vitro conditions. The aim of this study was to investigate the safety and efficacy of NVC-422 as local antibiofilm treatment in a sheep model of rhinosinusitis. After accessing and occluding frontal sinus ostia in 24 merino sheep via staged endoscopic procedures, S. aureus clinical isolate was instilled in frontal sinuses. Following biofilm formation, ostial obstruction was removed and sinuses irrigated with 0.1% and 0.5% NVC-422 in 5 mM acetate isotonic saline at pH 4.0. Sheep were monitored for adverse effects and euthanized 24 hours after treatment. Frontal sinuses were assessed for infection and changes in mucosa after the treatment. S. aureus biofilms were identified with Baclight-confocal scanning microscopy protocol and the biofilm biomass assayed by applying the COMSTAT2 program to recorded image stacks. After 2 irrigations with 0.1% NVC-422, S. aureus biofilm biomass was reduced when compared to control sinuses (p = 0.0001), though this effect was variable in samples. NVC-422 0.5% solution irrigations reduced biofilm even more significantly and consistently over all samples (p < 0.0001). NVC-422 0.5% was also more effective than 0.1% NVC-422, vehicle control, and normal saline sinus irrigations in reducing biofilm biomass (p < 0.05 for all subgroups). No adverse events were observed in sheep after sinus irrigations with 0.1% and 0.5% NVC-422 solutions. NVC-422 is an effective topical agent against S. aureus biofilms, with dose-dependent efficacy in this animal model of biofilm-associated sinusitis. Copyright © 2012 American Rhinologic Society-American Academy of Otolaryngic Allergy, LLC.

  8. Evaluation of the antifungal effect of EDTA, a metal chelator agent, on Candida albicans biofilm.

    PubMed

    Casalinuovo, I A; Sorge, R; Bonelli, G; Di Francesco, P

    2017-03-01

    Candida albicans biofilm is frequently found on artificial surfaces and the infections related to biofilm are difficult to eliminate, as they require the removal of artificial devices and treatment with antifungal drugs. Nowadays, fungal growth in biofilms is difficult to eradicate with conventional antifungal drugs such as fluconazole. Among chelating agents, disodium salt-Ethylene Diamine Tetraacetic Acid (EDTA) is known to have antifungal activity. In this study, we examined the in vitro activity of the EDTA and the antifungal drug fluconazole against C. albicans mature biofilm. C. albicans ATCC 20191, fluconazole-susceptible strain, was grown at an inoculum starter of 1 x 106 cells/ml for 72 h in 24-well microtiter plates and was further treated for 24 h with EDTA and/or fluconazole. Antifungal activities in biofilms were expressed as reduction in optical density (OD) determined by a 2,3-bis (2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide (XTT) colorimetric assay and compared to untreated biofilms. Colorimetric readings revealed that EDTA alone (at 25 and 2.5 mM) significantly reduced fungal metabolic activity in preformed biofilms. Also, EDTA combined with fluconazole significantly reduced the growth of biofilm when compared to biofilm treated with fluconazole alone (at 25 and 2.5 µg/ml). Our data suggest that the employment of EDTA or other chemicals destabilizers of the biofilm matrix, in combination with antifungal drugs, could lead to the development of new strategies for the management of infections associated to Candida biofilm. Another relevant result of our study suggests that the initial cell concentration, probably through mechanisms of quorum sensing, affects the cellular viability during the process of biofilm formation.

  9. Dissecting the regulation of bile-induced biofilm formation in Staphylococcus aureus.

    PubMed

    Ulluwishewa, Dulantha; Wang, Liang; Pereira, Callen; Flynn, Stephanie; Cain, Elizabeth; Stick, Stephen; Reen, F Jerry; Ramsay, Joshua P; O'Gara, Fergal

    2016-08-01

    Aspiration of bile into the cystic fibrosis (CF) lung has emerged as a prognostic factor for reduced microbial lung biodiversity and the establishment of often fatal, chronic pathogen infections. Staphylococcus aureus is one of the earliest pathogens detected in the lungs of children with CF, and once established as a chronic infection, strategies for its eradication become limited. Several lung pathogens are stimulated to produce biofilms in vitro in the presence of bile. In this study, we further investigated the effects of bile on S. aureus biofilm formation. Most clinical S. aureus strains and the laboratory strain RN4220 were stimulated to form biofilms with sub-inhibitory concentrations of bovine bile. Additionally, we observed bile-induced sensitivity to aminoglycosides, which we exploited in a bursa aurealis transposon screen to isolate mutants reduced in aminoglycoside sensitivity and augmented in bile-induced biofilm formation. We identified five mutants that exhibited hypersensitivity to bile with respect to bile-induced biofilm formation, three of which carried transposon insertions within gene clusters involved in wall teichoic acid (WTA) biosynthesis or transport. Strain TM4 carried an insertion between the divergently oriented tagH and tagG genes, which encode the putative WTA membrane translocation apparatus. Ectopic expression of tagG in TM4 restored a wild-type bile-induced biofilm response, suggesting that reduced translocation of WTA in TM4 induced sensitivity to bile and enhanced the bile-induced biofilm formation response. We propose that WTA may be important for protecting S. aureus against exposure to bile and that bile-induced biofilm formation may be an evolved response to protect cells from bile-induced cell lysis.

  10. Osmotic membrane bioreactor for phenol biodegradation under continuous operation.

    PubMed

    Praveen, Prashant; Loh, Kai-Chee

    2016-03-15

    Continuous phenol biodegradation was accomplished in a two-phase partitioning osmotic membrane bioreactor (TPPOMBR) system, using extractant impregnated membranes (EIM) as the partitioning phase. The EIMs alleviated substrate inhibition during prolonged operation at influent phenol concentrations of 600-2000mg/L, and also at spiked concentrations of 2500mg/L phenol restricted to 2 days. Filtration of the effluent through forward osmosis maintained high biomass concentration in the bioreactor and improved effluent quality. Steady state was reached in 5-6 days at removal rates varying between 2000 and 5500mg/L-day under various conditions. Due to biofouling and salt accumulation, the permeate flux varied from 1.2-7.2 LMH during 54 days of operation, while maintaining an average hydraulic retention time of 7.4h. A washing cycle, comprising 1h osmotic backwashing using 0.5M NaCl and 2h washing with water, facilitated biofilm removal from the membranes. Characterization of the extracellular polymeric substances (EPS) through FTIR showed peaks between 1700 and 1500cm(-1), 1450-1450cm(-1) and 1200-1000cm(-1), indicating the presence of proteins, phenols and polysaccharides, respectively. The carbohydrate to protein ratio in the EPS was estimated to be 0.3. These results indicate that TPPOMBR can be promising in continuous treatment of phenolic wastewater. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. A Sub-microsecond Pulsed Plasma Jet for Endodontic Biofilm Disinfection

    NASA Astrophysics Data System (ADS)

    Jiang, Chunqi; Schaudinn, Christoph; Jaramillo, David E.; Gundersen, Martin A.; Costerton, J. William

    A pulsed, tapered cylindrical plasma jet, several centimeter long and <2 mm in diameter, has been generated by a concentric tubular device for root canal disinfection. This plasma dental probe is typically powered with ˜100 ns, 1 kHz, multi-kilovolt electric pulses and filled with 5 SLPM (standard liter per minute) He/(1%)O2 flow. We report here an in vitro study of the antimicrobial effect of the room temperature plasma jet against monolayer Enterococcus faecalis biofilms on bovine dentins. Resultant colony-forming unit counts were associated with changes in bacterial cell morphology observed using scanning electron microscopy (SEM) following the treatment and control. Treatment of dentin discs cultivated with E. faecalis monolayer biofilms with the plasma (average power ≈ 1 W) for 5 min resulted in 92.4% kill (P < 0.0001). Severe disruption of the cell membranes was observed for the plasma treatment group, while the morphology of the cells remained intact for the negative control group. In addition, a pilot ex vivo test was conducted to examine the bactericidal effect of the plasma against saliva-derived biofilms cultivated in human root canals. Conspicuous biofilm disruption and cleared dentinal surfaces were observed in the canal after the plasma treatment for 5 min. We ­conclude that this non-thermal pulsed plasma-based technology is a potential ­alternative or supplement to existing protocols for root canal disinfection.

  12. Inquiry-based examination of chemical disruption of bacterial biofilms.

    PubMed

    Redelman, Carly V; Hawkins, Misty A W; Drumwright, Franklin R; Ransdell, Beverly; Marrs, Kathleen; Anderson, Gregory G

    2012-01-01

    Inquiry-based instruction in the sciences has been demonstrated as a successful educational strategy to use for both high school and college science classrooms. As participants in the NSF Graduate STEM Fellows in K-12 Education (GK-12) Program, we were tasked with creating novel inquiry-based activities for high school classrooms. As a way to introduce microbiology, molecular biology, ecology, and human health to students, we created a laboratory activity involving formation of biofilms composed of environmental bacteria from pond water and investigation into the resistance of these biofilms to antimicrobial agents. Two high schools participated in this study in different ways. Pike High School biology and advanced environmental science classrooms obtained pond water samples and grew biofilms from the bacteria in the pond water on plastic plates. They also observed killing of these biofilms by common household antimicrobial agents. As a senior capstone project, students at Arsenal Technical High School built on these research findings by isolating two different bacterial strains from the pond water and demonstrating the stimulatory effect of ethanol on biofilms formed by isolated bacterial strains. These activities were successful at introducing complex biological topics to high school students in a unique and exciting way. The students scored significantly higher on postactivity surveys compared with preactivity surveys that measured microbiology knowledge and experimental design knowledge. Furthermore, these projects seemed to elicit an excitement for science in the students who participated. Copyright © 2012 Wiley Periodicals, Inc.

  13. Revealing region-specific biofilm viscoelastic properties by means of a micro-rheological approach.

    PubMed

    Cao, Huayu; Habimana, Olivier; Safari, Ashkan; Heffernan, Rory; Dai, Yihong; Casey, Eoin

    2016-01-01

    Particle-tracking microrheology is an in situ technique that allows quantification of biofilm material properties. It overcomes the limitations of alternative techniques such as bulk rheology or force spectroscopy by providing data on region specific material properties at any required biofilm location and can be combined with confocal microscopy and associated structural analysis. This article describes single particle tracking microrheology combined with confocal laser scanning microscopy to resolve the biofilm structure in 3 dimensions and calculate the creep compliances locally. Samples were analysed from Pseudomonas fluorescens biofilms that were cultivated over two timescales (24 h and 48 h) and alternate ionic conditions (with and without calcium chloride supplementation). The region-based creep compliance analysis showed that the creep compliance of biofilm void zones is the primary contributor to biofilm mechanical properties, contributing to the overall viscoelastic character.

  14. Thioredoxin h regulates calcium dependent protein kinases in plasma membranes.

    PubMed

    Ueoka-Nakanishi, Hanayo; Sazuka, Takashi; Nakanishi, Yoichi; Maeshima, Masayoshi; Mori, Hitoshi; Hisabori, Toru

    2013-07-01

    Thioredoxin (Trx) is a key player in redox homeostasis in various cells, modulating the functions of target proteins by catalyzing a thiol-disulfide exchange reaction. Target proteins of cytosolic Trx-h of higher plants were studied, particularly in the plasma membrane, because plant plasma membranes include various functionally important protein molecules such as transporters and signal receptors. Plasma membrane proteins from Arabidopsis thaliana cell cultures were screened using a resin Trx-h1 mutant-immobilized, and a total of 48 candidate proteins obtained. These included two calcium-sensing proteins: a phosphoinositide-specific phospholipase 2 (AtPLC2) and a calcium-dependent protein kinase 21 (AtCPK21). A redox-dependent change in AtCPK21 kinase activity was demonstrated in vitro. Oxidation of AtCPK21 resulted in a decrease in kinase activity to 19% of that of untreated AtCPK21, but Trx-h1 effectively restored the activity to 90%. An intramolecular disulfide bond (Cys97-Cys108) that is responsible for this redox modulation was then identified. In addition, endogenous AtCPK21 was shown to be oxidized in vivo when the culture cells were treated with H2 O2 . These results suggest that redox regulation of AtCPK21 by Trx-h in response to external stimuli is important for appropriate cellular responses. The relationship between the redox regulation system and Ca(2+) signaling pathways is discussed. © 2013 The Authors. FEBS Journal published by John Wiley & Sons Ltd on behalf of FEBS.

  15. Ultrathin graphene oxide-based hollow fiber membranes with brush-like CO2-philic agent for highly efficient CO2 capture.

    PubMed

    Zhou, Fanglei; Tien, Huynh Ngoc; Xu, Weiwei L; Chen, Jung-Tsai; Liu, Qiuli; Hicks, Ethan; Fathizadeh, Mahdi; Li, Shiguang; Yu, Miao

    2017-12-13

    Among the current CO 2 capture technologies, membrane gas separation has many inherent advantages over other conventional techniques. However, fabricating gas separation membranes with both high CO 2 permeance and high CO 2 /N 2 selectivity, especially under wet conditions, is a challenge. In this study, sub-20-nm thick, layered graphene oxide (GO)-based hollow fiber membranes with grafted, brush-like CO 2 -philic agent alternating between GO layers are prepared by a facile coating process for highly efficient CO 2 /N 2 separation under wet conditions. Piperazine, as an effective CO 2 -philic agent, is introduced as a carrier-brush into the GO nanochannels with chemical bonding. The membrane exhibits excellent separation performance under simulated flue gas conditions with CO 2 permeance of 1,020 GPU and CO 2 /N 2 selectivity as high as 680, demonstrating its potential for CO 2 capture from flue gas. We expect this GO-based membrane structure combined with the facile coating process to facilitate the development of ultrathin GO-based membranes for CO 2 capture.

  16. Nitrate effects on chromate reduction in a methane-based biofilm.

    PubMed

    Zhong, Liang; Lai, Chun-Yu; Shi, Ling-Dong; Wang, Kai-Di; Dai, Yu-Jie; Liu, Yao-Wei; Ma, Fang; Rittmann, Bruce E; Zheng, Ping; Zhao, He-Ping

    2017-05-15

    The effects of nitrate (NO 3 - ) on chromate (Cr(VI)) reduction in a membrane biofilm reactor (MBfR) were studied when CH 4 was the sole electron donor supplied with a non-limiting delivery capacity. A high surface loading of NO 3 - gave significant and irreversible inhibition of Cr(VI) reduction. At a surface loading of 500 mg Cr/m 2 -d, the Cr(VI)-removal percentage was 100% when NO 3 - was absent (Stage 1), but was dramatically lowered to < 25% with introduction of 280 mg N m -2 -d NO 3 - (Stage 2). After ∼50 days operation in Stage 2, the Cr(VI) reduction recovered to only ∼70% in Stage 3, when NO 3 - was removed from the influent; thus, NO 3 - had a significant long-term inhibition effect on Cr(VI) reduction. Weighted PCoA and UniFrac analyses proved that the introduction of NO 3 - had a strong impact on the microbial community in the biofilms, and the changes possibly were linked to the irreversible inhibition of Cr(VI) reduction. For example, Meiothermus, the main genus involved in Cr(VI) reduction at first, declined with introduction of NO 3 - . The denitrifier Chitinophagaceae was enriched after the addition of NO 3 - , while Pelomonas became important when nitrate was removed, suggesting its potential role as a Cr(VI) reducer. Moreover, introducing NO 3 - led to a decrease in the number of genes predicted (by PICRUSt) to be related to chromate reduction, but genes predicted to be related to denitrification, methane oxidation, and fermentation increased. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Effects of oakmoss and its components on biofilm formation of Legionella pneumophila.

    PubMed

    Nomura, Harue; Isshiki, Yasunori; Sakuda, Keisuke; Sakuma, Katsuya; Kondo, Seiichi

    2013-01-01

    Oakmoss and its components are known as antibacterial agents, specifically against Legionella pneumophila. In the present study, we investigated the effects of oakmoss and its components (phenol, didepside and isochromen derivatives) on L. pneumophila biofilm formation, with particular reference to the bactericidal activity (minimum bactericidal concentration; MBC) of these components against the bacterial cells in the biofilm. Of the 20 compounds tested, two didepside derivatives and four phenol derivatives reduced biofilm formation by more than 50% of that observed for the control at their respective minimum inhibitory concentrations (1/2×MIC). The inhibitory activities of these compounds were either equivalent to or greater than that of the clarithromycin reference. Isochromen derivatives had no effect on biofilm formation. Analysis of bactericidal activity of didepside and isochromen derivatives revealed that three of four didepside derivatives and one of four isochromen derivatives exhibited high bactericidal activity (MBC: 32.0-74.7 µg/mL) against the L. pneumophila in the biofilm after 24 h or 48 h of co-incubation; the antibacterial activities of these compounds were almost equivalent to clarithromycin and chlorhexidine gluconate (MBC: 42.7-64.0 µg/mL) that were used as references. Thus, based on their anti-biofilm forming and bactericidal activities, didepside derivatives are considered to be good candidates for disinfectants against L. pneumophila.

  18. A Novel Soluble Peptide with pH-Responsive Membrane Insertion.

    PubMed

    Nguyen, Vanessa P; Alves, Daiane S; Scott, Haden L; Davis, Forrest L; Barrera, Francisco N

    2015-11-03

    Several diseases, such as cancer, are characterized by acidification of the extracellular environment. Acidosis can be employed as a target to specifically direct therapies to the diseased tissue. We have used first principles to design an acidity-triggered rational membrane (ATRAM) peptide with high solubility in solution that is able to interact with lipid membranes in a pH-dependent fashion. Biophysical studies show that the ATRAM peptide binds to the surface of lipid membranes at pH 8.0. However, acidification leads to the peptide inserting into the lipid bilayer as a transmembrane α-helix. The insertion of ATRAM into membranes occurs at a moderately acidic pH (with a pK of 6.5), similar to the extracellular pH found in solid tumors. Studies with human cell lines showed a highly efficient pH-dependent membrane targeting, without causing toxicity. Here we show that it is possible to rationally design a soluble peptide that selectively targets cell membranes in acidic environments.

  19. Sensing small molecule interactions with lipid membranes by local pH modulation.

    PubMed

    Huang, Da; Zhao, Tao; Xu, Wei; Yang, Tinglu; Cremer, Paul S

    2013-11-05

    Herein, we utilized a label-free sensing platform based on pH modulation to detect the interactions between tetracaine, a positively charged small molecule used as a local anesthetic, and planar supported lipid bilayers (SLBs). The SLBs were patterned inside a flow cell, allowing for various concentrations of tetracaine to be introduced over the surface in a buffer solution. Studies with membranes containing POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) yielded an equilibrium dissociation constant value of Kd = 180 ± 47 μm for this small molecule-membrane interaction. Adding cholesterol to the SLBs decreased the affinity between tetracaine and the bilayers, while this interaction tightened when POPE (1-hexadecanoyl-2-(9-Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine) was added. Studies were also conducted with three negatively charged membrane lipids, POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (sodium salt)), POPS (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-l-serine (sodium salt)), and ganglioside GM1. All three measurements gave rise to a similar tightening of the apparent Kd value compared with pure POPC membranes. The lack of chemical specificity with the identity of the negatively charged lipid indicated that the tightening was largely electrostatic. Through a direct comparison with ITC measurements, it was found that the pH modulation sensor platform offers a facile, inexpensive, highly sensitive, and rapid method for the detection of interactions between putative drug candidates and lipid bilayers. As such, this technique may potentially be exploited as a screen for drug development and analysis.

  20. Hibiscus sabdariffa extract inhibits in vitro biofilm formation capacity of Candida albicans isolated from recurrent urinary tract infections

    PubMed Central

    Alshami, Issam; Alharbi, Ahmed E

    2014-01-01

    Objective To explore the prevention of recurrent candiduria using natural based approaches and to study the antimicrobial effect of Hibiscus sabdariffa (H. sabdariffa) extract and the biofilm forming capacity of Candida albicans strains in the present of the H. sabdariffa extract. Methods In this particular study, six strains of fluconazole resistant Candida albicans isolated from recurrent candiduria were used. The susceptibility of fungal isolates, time-kill curves and biofilm forming capacity in the present of the H. sabdariffa extract were determined. Results Various levels minimum inhibitory concentration of the extract were observed against all the isolates. Minimum inhibitory concentration values ranged from 0.5 to 2.0 mg/mL. Time-kill experiment demonstrated that the effect was fungistatic. The biofilm inhibition assay results showed that H. sabdariffa extract inhibited biofilm production of all the isolates. Conclusions The results of the study support the potential effect of H. sabdariffa extract for preventing recurrent candiduria and emphasize the significance of the plant extract approach as a potential antifungal agent. PMID:25182280

  1. Basolateral membrane Na/base cotransport is dependent on CO2/HCO3 in the proximal convoluted tubule

    PubMed Central

    1987-01-01

    The mechanism of basolateral membrane base transport was examined in the in vitro microperfused rabbit proximal convoluted tubule (PCT) in the absence and presence of ambient CO2/HCO3- by means of the microfluorometric measurement of cell pH. The buffer capacity of the cells measured using rapid NH3 washout was 42.8 +/- 5.6 mmol.liter-1.pH unit-1 in the absence and 84.6 +/- 7.3 mmol.liter-1.pH unit-1 in the presence of CO2/HCO3-. In the presence of CO2/HCO3-, lowering peritubular pH from 7.4 to 6.8 acidified the cell by 0.30 pH units and lowering peritubular Na from 147 to 0 mM acidified the cell by 0.25 pH units. Both effects were inhibited by peritubular 4-acetamido-4'- isothiocyanostilbene-2,2'-disulfonate (SITS). In the absence of exogenous CO2/HCO3-, lowering peritubular pH from 7.4 to 6.8 acidified the cell by 0.25 pH units and lowering peritubular Na from 147 to 0 mM decreased cell pH by 0.20 pH units. Lowering bath pH from 7.4 to 6.8 induced a proton flux of 643 +/- 51 pmol.mm-1.min-1 in the presence of exogenous CO2/HCO3- and 223 +/- 27 pmol.mm-1.min-1 in its absence. Lowering bath Na from 147 to 0 mM induced proton fluxes of 596 +/- 77 pmol.mm-1.min-1 in its absence. The cell acidification induced by lowering bath pH or bath Na in the absence of CO2/HCO3- was inhibited by peritubular SITS or by acetazolamide, whereas peritubular amiloride had no effect. In the absence of exogenous CO2/HCO3-, cyanide blocked the cell acidification induced by bath Na removal, but was without effect in the presence of exogenous CO2/HCO3-. We reached the following conclusions. (a) The basolateral Na/base n greater than 1 cotransporter in the rabbit PCT has an absolute requirement for CO2/HCO3-. (b) In spite of this CO2 dependence, in the absence of exogenous CO2/HCO3-, metabolically produced CO2/HCO3- is sufficient to keep the transporter running at 30% of its control rate in the presence of ambient CO2/HCO3- . (c) There is no apparent amiloride-sensitive Na/H antiporter on

  2. [3H]-nitrendipine binding in membranes obtained from hypoxic and reoxygenated heart.

    PubMed

    Matucci, R; Bennardini, F; Sciammarella, M L; Baccaro, C; Stendardi, I; Franconi, F; Giotti, A

    1987-04-01

    We compared the binding properties of [3H]-nitrendipine in heart membranes from normal guinea-pig heart and from hypoxic or hypoxic and reoxygenated heart. The [3H]-nitrendipine binds a single class of high capacity (Bmax 667.2 +/- 105.2) with high affinity (KD 0.14 +/- 0.02) binding sites. By contrast, in membranes of hypoxic and reoxygenated heart the Bmax decreases significantly while it remains unaffected during hypoxia. Xanthinoxidase activity is increased in hypoxic-reoxygenated hearts.

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

    PubMed

    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.

  4. Early detection of Candida albicans biofilms at porous electrodes.

    PubMed

    Congdon, Robert B; Feldberg, Alexander S; Ben-Yakar, Natalie; McGee, Dennis; Ober, Christopher; Sammakia, Bahgat; Sadik, Omowunmi A

    2013-02-15

    We describe the development of an electrochemical sensor for early detection of biofilm using Candida albicans. The electrochemical sensor used the ability of biofilms to accept electrons from redox mediators relative to the number of metabolically active cells present. Cyclic voltammetry and differential pulse voltammetry techniques were used to monitor the redox reaction of K(3)Fe(CN)(6) at porous reticulated vitreous carbon (RVC) (238.7 cm(2)) working electrodes versus Ag/AgCl reference. A shift in the peak potential occurred after 12 h of film growth, which is attributed to the presence of C. albicans. Moreover, the intensity of the ferricyanide reduction peak first increased as C. albicans deposited onto the porous electrodes at various growth times. The peak current subsequently decreased at extended periods of growth of 48 h. The reduction in peak current was attributed to the biofilm reaching its maximum growth thickness, which correlated with the maximum number of metabolically active cells. The observed diffusion coefficients for the bare RVC and biofilm-coated electrodes were 2.2 × 10(-3) and 7.0 × 10(-6) cm(2)/s, respectively. The increase in diffusivity from the bare electrode to the biofilm-coated electrode indicated some enhancement of electron transfer mediated by the biofilm to the porous electrode. Verification of the growth of biofilm was achieved using scanning electron microcopy and laser scanning confocal imaging microscopy. Validation with conventional plating techniques confirmed that the correlation (R(2) = 0.9392) could be achieved between the electrochemical sensors data and colony-forming units. Copyright © 2012 Elsevier Inc. All rights reserved.

  5. Monitoring in Real Time the Formation and Removal of Biofilms from Clinical Related Pathogens Using an Impedance-Based Technology

    PubMed Central

    Gutiérrez, Diana; Hidalgo-Cantabrana, Claudio; Rodríguez, Ana; García, Pilar

    2016-01-01

    Bacteria found in diverse ecosystems grow in a community of aggregated cells that favors their survival and colonization. Different extracellular polymeric substances are used to entrap this multispecies community forming a biofilm, which can be associated to biotic and abiotic surfaces. This widespread and successful way of bacterial life, however, can lead to negative effects for human activity since many pathogen and spoiling bacteria form biofilms which are not easy to eradicate. Therefore, the search for novel anti-biofilm bio-active molecules is a very active research area for which simple, reliable, and fast screening methods are demanded. In this work we have successfully validated an impedance-based method, initially developed for the study of adherent eukaryotic cells, to monitor the formation of single-species biofilms of three model bacteria in real time. The xCelligence real time cell analyzer (RTCA) equipment uses specific microtiter E-plates coated with gold-microelectrodes that detect the attachment of adherent cells, thus modifying the impedance signal. In the current study, this technology allowed the distinction between biofilm-producers and non-producers of Staphylococcus aureus and Staphylococcus epidermidis, as well as the formation of Streptococcus mutans biofilms only when sucrose was present in the culture medium. Besides, different impedance values permitted discrimination among the biofilm-producing strains tested regardless of the nature of the polymeric biofilm matrix. Finally, we have continuously monitored the inhibition of staphylococcal biofilm formation by the bacteriophage phi-IPLA7 and the bacteriophage-encoded endolysin LysH5, as well as the removal of a preformed biofilm by this last antimicrobial treatment. Results observed with the impedance-based method showed high correlation with those obtained with standard approaches, such as crystal violet staining and bacteria enumeration, as well as with those obtained upon other

  6. Monitoring in Real Time the Formation and Removal of Biofilms from Clinical Related Pathogens Using an Impedance-Based Technology.

    PubMed

    Gutiérrez, Diana; Hidalgo-Cantabrana, Claudio; Rodríguez, Ana; García, Pilar; Ruas-Madiedo, Patricia

    2016-01-01

    Bacteria found in diverse ecosystems grow in a community of aggregated cells that favors their survival and colonization. Different extracellular polymeric substances are used to entrap this multispecies community forming a biofilm, which can be associated to biotic and abiotic surfaces. This widespread and successful way of bacterial life, however, can lead to negative effects for human activity since many pathogen and spoiling bacteria form biofilms which are not easy to eradicate. Therefore, the search for novel anti-biofilm bio-active molecules is a very active research area for which simple, reliable, and fast screening methods are demanded. In this work we have successfully validated an impedance-based method, initially developed for the study of adherent eukaryotic cells, to monitor the formation of single-species biofilms of three model bacteria in real time. The xCelligence real time cell analyzer (RTCA) equipment uses specific microtiter E-plates coated with gold-microelectrodes that detect the attachment of adherent cells, thus modifying the impedance signal. In the current study, this technology allowed the distinction between biofilm-producers and non-producers of Staphylococcus aureus and Staphylococcus epidermidis, as well as the formation of Streptococcus mutans biofilms only when sucrose was present in the culture medium. Besides, different impedance values permitted discrimination among the biofilm-producing strains tested regardless of the nature of the polymeric biofilm matrix. Finally, we have continuously monitored the inhibition of staphylococcal biofilm formation by the bacteriophage phi-IPLA7 and the bacteriophage-encoded endolysin LysH5, as well as the removal of a preformed biofilm by this last antimicrobial treatment. Results observed with the impedance-based method showed high correlation with those obtained with standard approaches, such as crystal violet staining and bacteria enumeration, as well as with those obtained upon other

  7. Synthetic Arabinomannan Heptasaccharide Glycolipids Inhibit Biofilm Growth and Augment Isoniazid Effects in Mycobacterium smegmatis.

    PubMed

    Maiti, Krishnagopal; Syal, Kirtimaan; Chatterji, Dipankar; Jayaraman, Narayanaswamy

    2017-10-05

    Biofilm formation, involving attachment to an adherent surface, is a critical survival strategy of mycobacterial colonies in hostile environmental conditions. Here we report the synthesis of heptasaccharide glycolipids based on mannopyranoside units anchored on to a branched arabinofuranoside core. Two types of glycolipids-2,3-branched and 2,5-branched-were synthesized and evaluated for their efficacies in inhibiting biofilm growth by the non-pathogenic mycobacterium variant Mycobacterium smegmatis. Biofilm formation was inhibited at a minimum biofilm growth inhibition concentration (MBIC) of 100 μg mL -1 in the case of the 2,5-branched heptasaccharide glycolipid. Further, we were able to ascertain that a combination of the drug isoniazid with the branched heptasaccharide glycolipid (50 μg mL -1 ) potentiates the drug, making it three times more effective, with an improved MBIC of 30 μg mL -1 . These studies establish that synthetic glycolipids not only act as inhibitors of biofilm growth, but also provide a synergistic effect when combined with significantly lowered concentrations of isoniazid to disrupt the biofilm structures of the mycobacteria. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. The extracellular matrix of Staphylococcus aureus biofilms comprises cytoplasmic proteins that associate with the cell surface in response to decreasing pH.

    PubMed

    Foulston, Lucy; Elsholz, Alexander K W; DeFrancesco, Alicia S; Losick, Richard

    2014-09-02

    Biofilm formation by Staphylococcus aureus involves the formation of an extracellular matrix, but the composition of this matrix has been uncertain. Here we report that the matrix is largely composed of cytoplasmic proteins that reversibly associate with the cell surface in a manner that depends on pH. We propose a model for biofilm formation in which cytoplasmic proteins are released from cells in stationary phase. These proteins associate with the cell surface in response to decreasing pH during biofilm formation. Rather than utilizing a dedicated matrix protein, S. aureus appears to recycle cytoplasmic proteins that moonlight as components of the extracellular matrix. Staphylococcus aureus is a leading cause of multiantibiotic-resistant nosocomial infections and is often found growing as a biofilm in catheters and chronic wounds. Biofilm formation is an important pathogenicity strategy that enhances resistance to antimicrobials, thereby limiting treatment options and ultimately contributing to increased morbidity and mortality. Cells in a biofilm are held together by an extracellular matrix that consists in whole or in part of protein, but the nature of the proteins in the S. aureus matrix is not well understood. Here we postulate that S. aureus recycles proteins from the cytoplasm to form the extracellular matrix. This strategy, of cytoplasmic proteins moonlighting as matrix proteins, could allow enhanced flexibility and adaptability for S. aureus in forming biofilms under infection conditions and could promote the formation of mixed-species biofilms in chronic wounds. Copyright © 2014 Foulston et al.

  9. Metabolic activity of Streptococcus mutans biofilms and gene expression during exposure to xylitol and sucrose.

    PubMed

    Decker, Eva-Maria; Klein, Christian; Schwindt, Dimitri; von Ohle, Christiane

    2014-12-01

    The objective of the study was to analyse Streptococcus mutans biofilms grown under different dietary conditions by using multifaceted methodological approaches to gain deeper insight into the cariogenic impact of carbohydrates. S. mutans biofilms were generated during a period of 24 h in the following media: Schaedler broth as a control medium containing endogenous glucose, Schaedler broth with an additional 5% sucrose, and Schaedler broth supplemented with 1% xylitol. The confocal laser scanning microscopy (CLSM)-based analyses of the microbial vitality, respiratory activity (5-cyano-2,3-ditolyl tetrazolium chloride, CTC) and production of extracellular polysaccharides (EPS) were performed separately in the inner, middle and outer biofilm layers. In addition to the microbiological sample testing, the glucose/sucrose consumption of the biofilm bacteria was quantified, and the expression of glucosyltransferases and other biofilm-associated genes was investigated. Xylitol exposure did not inhibit the viability of S. mutans biofilms, as monitored by the following experimental parameters: culture growth, vitality, CTC activity and EPS production. However, xylitol exposure caused a difference in gene expression compared to the control. GtfC was upregulated only in the presence of xylitol. Under xylitol exposure, gtfB was upregulated by a factor of 6, while under sucrose exposure, it was upregulated by a factor of three. Compared with glucose and xylitol, sucrose increased cell vitality in all biofilm layers. In all nutrient media, the intrinsic glucose was almost completely consumed by the cells of the S. mutans biofilm within 24 h. After 24 h of biofilm formation, the multiparametric measurements showed that xylitol in the presence of glucose caused predominantly genotypic differences but did not induce metabolic differences compared to the control. Thus, the availability of dietary carbohydrates in either a pure or combined form seems to affect the

  10. Metabolic activity of Streptococcus mutans biofilms and gene expression during exposure to xylitol and sucrose

    PubMed Central

    Decker, Eva-Maria; Klein, Christian; Schwindt, Dimitri; von Ohle, Christiane

    2014-01-01

    The objective of the study was to analyse Streptococcus mutans biofilms grown under different dietary conditions by using multifaceted methodological approaches to gain deeper insight into the cariogenic impact of carbohydrates. S. mutans biofilms were generated during a period of 24 h in the following media: Schaedler broth as a control medium containing endogenous glucose, Schaedler broth with an additional 5% sucrose, and Schaedler broth supplemented with 1% xylitol. The confocal laser scanning microscopy (CLSM)-based analyses of the microbial vitality, respiratory activity (5-cyano-2,3-ditolyl tetrazolium chloride, CTC) and production of extracellular polysaccharides (EPS) were performed separately in the inner, middle and outer biofilm layers. In addition to the microbiological sample testing, the glucose/sucrose consumption of the biofilm bacteria was quantified, and the expression of glucosyltransferases and other biofilm-associated genes was investigated. Xylitol exposure did not inhibit the viability of S. mutans biofilms, as monitored by the following experimental parameters: culture growth, vitality, CTC activity and EPS production. However, xylitol exposure caused a difference in gene expression compared to the control. GtfC was upregulated only in the presence of xylitol. Under xylitol exposure, gtfB was upregulated by a factor of 6, while under sucrose exposure, it was upregulated by a factor of three. Compared with glucose and xylitol, sucrose increased cell vitality in all biofilm layers. In all nutrient media, the intrinsic glucose was almost completely consumed by the cells of the S. mutans biofilm within 24 h. After 24 h of biofilm formation, the multiparametric measurements showed that xylitol in the presence of glucose caused predominantly genotypic differences but did not induce metabolic differences compared to the control. Thus, the availability of dietary carbohydrates in either a pure or combined form seems to affect the

  11. Streptococcus pneumoniae Eradicates Preformed Staphylococcus aureus Biofilms through a Mechanism Requiring Physical Contact.

    PubMed

    Khan, Faidad; Wu, Xueqing; Matzkin, Gideon L; Khan, Mohsin A; Sakai, Fuminori; Vidal, Jorge E

    2016-01-01

    Staphylococcus aureus (Sau) strains are a main cause of disease, including nosocomial infections which have been linked to the production of biofilms and the propagation of antibiotic resistance strains such as methicillin-resistant Staphylococcus aureus (MRSA). A previous study found that Streptococcus pneumoniae (Spn) strains kill planktonic cultures of Sau strains. In this work, we have further evaluated in detail the eradication of Sau biofilms and investigated ultrastructural interactions of the biofilmicidal effect. Spn strain D39, which produces the competence stimulating peptide 1 (CSP1), reduced Sau biofilms within 8 h of inoculation, while TIGR4, producing CSP2, eradicated Sau biofilms and planktonic cells within 4 h. Differences were not attributed to pherotypes as other Spn strains producing different pheromones eradicated Sau within 4 h. Experiments using Transwell devices, which physically separated both species growing in the same well, demonstrated that direct contact between Spn and Sau was required to efficiently eradicate Sau biofilms and biofilm-released planktonic cells. Physical contact-mediated killing of Sau was not related to production of hydrogen peroxide as an isogenic TIGR4Δ spx B mutant eradicated Sau bacteria within 4 h. Confocal micrographs confirmed eradication of Sau biofilms by TIGR4 and allowed us to visualize ultrastructural point of contacts between Sau and Spn. A time-course study further demonstrated spatial colocalization of Spn chains and Sau tetrads as early as 30 min post-inoculation (Pearson's coefficient >0.72). Finally, precolonized biofilms produced by Sau strain Newman, or MRSA strain USA300, were eradicated by mid-log phase cultures of washed TIGR4 bacteria within 2 h post-inoculation. In conclusion, Spn strains rapidly eradicate pre-colonized Sau aureus biofilms, including those formed by MRSA strains, by a mechanism(s) requiring bacterium-bacterium contact, but independent from the production of hydrogen peroxide.

  12. Na+/H+ exchange activity in the plasma membrane of Arabidopsis.

    PubMed

    Qiu, Quan-Sheng; Barkla, Bronwyn J; Vera-Estrella, Rosario; Zhu, Jian-Kang; Schumaker, Karen S

    2003-06-01

    In plants, Na+/H+ exchangers in the plasma membrane are critical for growth in high levels of salt, removing toxic Na+ from the cytoplasm by transport out of the cell. The molecular identity of a plasma membrane Na+/H+ exchanger in Arabidopsis (SOS1) has recently been determined. In this study, immunological analysis provided evidence that SOS1 localizes to the plasma membrane of leaves and roots. To characterize the transport activity of this protein, purified plasma membrane vesicles were isolated from leaves of Arabidopsis. Na+/H+ exchange activity, monitored as the ability of Na to dissipate an established pH gradient, was absent in plants grown without salt. However, exchange activity was induced when plants were grown in 250 mm NaCl and increased with prolonged salt exposure up to 8 d. H+-coupled exchange was specific for Na, because chloride salts of other monovalent cations did not dissipate the pH gradient. Na+/H+ exchange activity was dependent on Na (substrate) concentration, and kinetic analysis indicated that the affinity (apparent Km) of the transporter for Na+ is 22.8 mm. Data from two experimental approaches supports electroneutral exchange (one Na+ exchanged for one proton): (a) no change in membrane potential was measured during the exchange reaction, and (b) Na+/H+ exchange was unaffected by the presence or absence of a membrane potential. Results from this research provide a framework for future studies into the regulation of the plant plasma membrane Na+/H+ exchanger and its relative contribution to the maintenance of cellular Na+ homeostasis during plant growth in salt.

  13. [Formation of the Pseudomonas aeruginosa PAO1 biofilms in the presence of hydrogen peroxide; the effect of the AiiA gene].

    PubMed

    Pliuta, V A; Andreenko, Iu V; Kuznetsov, A E; Khmel', I A

    2013-01-01

    In the natural ecosystems, most bacteria exist as specifically organized biofilms attached to various surfaces; the biofilms have a complex architecture and are surrounded by an exopolymeric matrix. The bacteria in the biofilms are extremely resistant to antibacterial agents. The ability of the pathogenic bacteria to produce biofilms causes serious problems in medicine. Therefore, the study of the action of different compounds with antibacterial activity is of great interest. In this work, we studied the effect of the hydrogen peroxide (H2O2) on the formation of biofilms by Pseudomonas aeruginosa PAO1. It was shown that H2O2 in concentrations that do not suppress bacterial growth (or suppress it only weakly) stimulates the formation of the biofilms. At higher concentrations, H2O2 inhibits the formation of the biofilms. In order to determine if the stimulation of the biofilm formation depends on Quorum Sensing (QS) regulation, the plasmid pME6863 containing the heterologous gene aiiA encoding the N-acyl-homoserine lactonase AiiA was introduced into P. aeruginosa PAO1. The synthesis by cells of this enzyme degrading N-acyl-homoserine lactones (AHL), signaling molecules of the QS systems, led to the absence of the stimulation of the biofilm formation by the action of H2O2. This fact indicates that the stimulation of the biofilm formation in the presence of H2O2 depends on the functioning of the QS systems of the gene expression regulation of P. aeruginosa PAO1.

  14. Comparison between a moving bed membrane bioreactor and a conventional membrane bioreactor on organic carbon and nitrogen removal.

    PubMed

    Yang, Shuai; Yang, Fenglin; Fu, Zhimin; Lei, Ruibo

    2009-04-01

    A membrane bioreactor filled with carriers instead of activated sludge named a moving bed membrane bioreactor (MBMBR) was investigated for simultaneously removing organic carbon and nitrogen in wastewater. Its performance was compared with a conventional membrane bioreactor (CMBR) at various influent COD/TN ratios of 8.9-22.1. The operational parameters were optimized to increase the treatment efficiency. COD removal efficiency averaged at 95.6% and 96.2%, respectively, for MBMBR and CMBR during the 4 months experimental period. The MBMBR system demonstrated good performance on nitrogen removal at different COD/TN ratios. When COD/TN was 8.9 and the total nitrogen (TN) load was 7.58 mg/l h, the TN and ammonium nitrogen removal efficiencies of the MBMBR were maintained over 70.0% and 80.0%, respectively, and the removed total nitrogen (TN) load reached to 5.31 mg/l h. Multifunctional microbial reactions in the carrier, such as simultaneous nitrification and denitrification (SND), play important roles in nitrogen removal. In comparison, the CMBR did not perform so well. Its TN removal was not stable, and the removed total nitrogen (TN) load was only 1.02 mg/l h at COD/TN ratio 8.9. The specific oxygen utilization rate (SOUR) showed that the biofilm has a better microbial activity than an activated sludge. Nevertheless, the membrane fouling behavior was more severe in the MBMBR than in the CMBR due to a thick and dense cake layer formed on the membrane surface, which was speculated to be caused by the filamentous bacteria in the MBMBR.

  15. Aminoglycoside inhibition of Staphylococcus aureus biofilm formation is nutrient dependent

    PubMed Central

    Hess, Donavon J.; Wells, Carol L.

    2014-01-01

    Biofilms represent microbial communities, encased in a self-produced matrix or extracellular polymeric substance. Microbial biofilms are likely responsible for a large proportion of clinically significant infections and the multicellular nature of biofilm existence has been repeatedly associated with antibiotic resistance. Classical in vitro antibiotic-susceptibility testing utilizes artificial growth media and planktonic microbes, but this method may not account for the variability inherent in environments subject to biofilm growth in vivo. Experiments were designed to test the hypothesis that nutrient concentration can modulate the antibiotic susceptibility of Staphylococcus aureus biofilms. Developing S. aureus biofilms initiated on surgical sutures, and in selected experiments planktonic cultures, were incubated for 16 h in 66 % tryptic soy broth, 0.2 % glucose (1× TSBg), supplemented with bactericidal concentrations of gentamicin, streptomycin, ampicillin or vancomycin. In parallel experiments, antibiotics were added to growth medium diluted one-third (1/3× TSBg) or concentrated threefold (3× TSBg). Following incubation, viable bacteria were enumerated from planktonic cultures or suture sonicates, and biofilm biomass was assayed using spectrophotometry. Interestingly, bactericidal concentrations of gentamicin (5 µg gentamicin ml−1) and streptomycin (32 µg streptomycin ml−1) inhibited biofilm formation in samples incubated in 1/3× or 1× TSBg, but not in samples incubated in 3× TSBg. The nutrient dependence of aminoglycoside susceptibility is not only associated with biofilm formation, as planktonic cultures incubated in 3× TSBg in the presence of gentamicin also showed antibiotic resistance. These findings appeared specific for aminoglycosides because biofilm formation was inhibited in all three growth media supplemented with bactericidal concentrations of the cell wall-active antibiotics, ampicillin and vancomycin. Additional experiments

  16. Baccharis dracunculifolia-based mouthrinse alters the exopolysaccharide structure in cariogenic biofilms.

    PubMed

    Aires, Carolina P; Sassaki, Guilherme L; Santana-Filho, Arquimedes P; Spadaro, Augusto C C; Cury, Jaime A

    2016-03-01

    Baccharis dracunculifolia is a native plant from Brazil with antimicrobial activity. The purpose of this study was to investigate whether a B. dracunculifolia-based mouthrinse (Bd) changes the structure of insoluble exopolysaccharides (IEPS) in Streptococcus mutans UA159 cariogenic biofilm. Biofilms were grown on glass slides and treated with Bd, its vehicle (VC), chlorhexidine digluconate (CHX), or saline solution (NaCl). Among the treatments, only CHX significantly reduced the biofilm biomass and bacterial viability (p<0.05). Gas chromatography-mass spectrometry and nuclear magnetic resonance analyses revealed that IEPS from the four biofilm samples were α- glucans containing different proportions of (1→6) and (1→3) glycosidic linkages. The structural differences among the four IEPS were compared by principal component analysis (PCA). PCA analysis indicated that IEPS from VC- and NaCl-treated biofilms were structurally similar to each other. Compared with the control, IEPS from Bd- and CHX-treated biofilms were structurally different and had distinct chemical profiles. In summary, the fact that Bd changed the IEPS chemical composition indicates that this mouthrinse may affect the cariogenic properties of the S. mutans biofilm formed. Copyright © 2015. Published by Elsevier B.V.

  17. Precisely Controlling Nanochannels of Graphene Oxide Membranes through Lignin-Based Cation Decoration for Dehydration of Biofuels.

    PubMed

    Guan, Kecheng; Liu, Quan; Ji, Yufan; Zhang, Mengchen; Wu, Yulin; Zhao, Jing; Liu, Gongping; Jin, Wanqin

    2018-05-07

    Lignin-based cations introduced into graphene oxide (GO) have been found to bring about stabilization of the nanostructure and the active sites and to give rise to various interactions for subsequent modification with polyelectrolyte and nanospacers, with a view to precisely controlling the nanochannels of the GO-based membranes. The resulting membranes exhibited excellent performance in biofuel dehydration with water flux of 4000-6000 g m -2h -1 , which exceeds that of the state-of-the-art polymeric and GO-based membranes. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Short communication: Roles of outer membrane protein W (OmpW) on survival and biofilm formation of Cronobacter sakazakii under neomycin sulfate stress.

    PubMed

    Ye, Yingwang; Ling, Na; Gao, Jina; Zhang, Maofeng; Zhang, Xiyan; Tong, Liaowang; Ou, Dexin; Wang, Yaping; Zhang, Jumei; Wu, Qingping

    2018-04-01

    Cronobacter sakazakii is associated with severe infections including sepsis, neonatal meningitis, and necrotizing enterocolitis. Antibiotic resistance in Cronobacter species has been documented in recent years, but the genes involved in resistance in Cronobacter strains are poorly understood. In this study, we determined the role of outer membrane protein W (OmpW) on survival rates, morphologic changes, and biofilm formation between wild type (WT) and an OmpW mutant strain (ΔOmpW) under neomycin sulfate stress. Results indicated that the survival rates of ΔOmpW were significantly reduced after half minimum inhibitory concentration (½ MIC) treatment compared with the WT strain. Filamentation of C. sakazakii cells was observed after ½ MIC treatment in WT and ΔOmpW, and morphologic injury, including cell disruption and leakage of cells, was more predominant in ΔOmpW. Under ½ MIC stress, the biofilms of WT and ΔOmpW were significantly decreased, but decreasing rates of biofilm formation in mutant strain were more predominant compared with WT strain. This is the first report to determine the role of OmpW on survival, morphological changes, and biofilm formation in C. sakazakii under neomycin sulfate stress. The findings indicated that OmpW contributed to survival and reduction of morphological injury under neomycin sulfate stress. In addition, enhancing biofilm formation in ΔOmpW may be an alternative advantage for adaptation to neomycin sulfate stress. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  19. Effect of chlorine treatment on inhibition of E. coli serogroup O2 incorporation into 7-day-old biofilm on polyvinylchloride surface.

    PubMed

    Maharjan, P; Dey, S; Huff, G; Zhang, W; Phillips, G K; Watkins, S

    2017-08-01

    Poultry waterlines are constructed using polyvinylchloride (PVC) material on which bacterial biofilm can easily form. Biofilm can harbor pathogens including avian pathogenic E. coli (APEC) strains. An in vitro evaluation was performed to determine if E. coli sero group O2 (avian pathogenic) could attach on a PVC surface that had pre-formed biofilm and if this phenomenon could be affected when water was treated with chlorine. Initially, biofilm growth was induced in PVC test coupons (15.16 cm2) for a 7-day period mimicking the waterline scenario in the first wk of poultry brooding; and then this biofilm was challenged with E. coli O2 seeded water in presence/absence of chlorine treatment. After rinsing, test coupons were sampled for bacterial (APC) and E. coli O2 enumeration at various occasions post seeding the pathogen and chlorine treatment. Day 7 APC recovered from coupons was 4.35 log10 cfu/cm2 in trial 1 and 3.66 log10 cfu/cm2 in trial 2. E. coli O2 was not recovered from chlorine treated test coupons (P < 0.05), whereas it was retrieved from untreated coupons (untreated contained > 3 log10 cfu/cm2 in trial 1 and > 2 log10 cfu/cm2 in trial 2). This study suggests that E. coli O2 can incorporate into pre-formed biofilm on a PVC surface within 24 h if water sanitation is not present, and the attachment time of the pathogen can prolong in the absence of already formed biofilm. © 2017 Poultry Science Association Inc.

  20. Addition of DNase Improves the In Vitro Activity of Antifungal Drugs against Candida albicans Biofilms

    PubMed Central

    Martins, Margarida; Henriques, Mariana; Lopez-Ribot, José L.; Oliveira, Rosário

    2011-01-01

    SUMMARY Background Cells within Candida albicans biofilms display decreased susceptibility to most clinically used antifungal agents. We recently demonstrated that extracellular DNA (eDNA) plays an important role in biofilm integrity, as a component of the biofilm matrix. Objective To gain insight into the contributions of eDNA to C. albicans biofilms antifungal susceptibility by the investigation of the impact of the combined use of deoxyribonuclease I (DNase) and antifungals to treat biofilms. Methods C. albicans biofilms were formed using a simple and reproducible 96-well plate-based method. The activity of the combined use of 0.13 mg l−1 DNase and antifungals was estimated by the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay, and total viable counts. Results and Conclusions Here we report the improved efficacy of amphotericin B when in combination with DNase against C. albicans biofilms, as assessed by XTT readings and viable counts. Furthermore, although DNase increased the efficacy of caspofungin in the reduction of mitochondrial activity, no changes were observed in terms of culturable cells. DNase did not affect biofilm cells susceptibility to fluconazole. This work suggests that agents that target processes affecting the biofilm structural integrity may have potential use as adjuvants of a catheter–lock therapy. PMID:21668524

  1. Alkaline Peroxides Versus Sodium Hypochlorite for Removing Denture Biofilm: a Crossover Randomized Trial.

    PubMed

    Peracini, Amanda; Regis, Rômulo Rocha; Souza, Raphael Freitas de; Pagnano, Valéria Oliveira; Silva, Cláudia Helena Lovato da; Paranhos, Helena de Freitas Oliveira

    2016-01-01

    This study evaluated the efficacy of cleanser solutions on denture biofilm removal by a crossover randomized clinical trial. Thirty two edentulous patients were instructed to brush their dentures (specific brush and liquid soap) three times a day (after breakfast, lunch and dinner) and to soak them (≥ 8 h) in: (C) control -water; (AP): alkaline peroxide; or (SH) 0.5% sodium hypochlorite. Each solution was used for 21 days (three cycles of 7 days). At the end of each cycle, the inner surfaces of maxillary dentures were disclosed (1% neutral red) and photographed (HX1 - Sony). Areas (total and stained biofilm) were measured (Image Tool software) and the percentage of biofilm calculated as the ratio between the area of the biofilm multiplied by 100 and total surface area of the internal base of the denture. Data were compared by means of generalized estimating equation (α=5%) and multiple comparisons (Bonferroni; α=1.67%). Immersion in SH reduced biofilm (%) (8.3 ± 13.3B) compared to C (18.2 ± 14.9A) and AP (18.2 ± 16.6A). The 0.5% sodium hypochlorite solution was the most efficacious for biofilm removal. Alkaline peroxides may not lead to further biofilm removal in patients with adequate denture maintenance habits.

  2. Fliposomes: pH-triggered conformational flip of new trans-2-aminocyclohexanol-based amphiphiles causes instant cargo release in liposomes.

    PubMed

    Liu, Xin; Zheng, Yu; Samoshina, Nataliya M; Franz, Andreas H; Guo, Xin; Samoshin, Vyacheslav V

    2012-12-01

    A new type of pH-sensitive liposomes (fliposomes) was designed based on the amphiphiles that are able to perform a pH-triggered conformational flip (flipids). This flip disrupts the liposome membrane and causes rapid release of the liposome cargo, specifically in response to lowered pH. The flipids (1) and (2) are equipped with a trans-2-aminocyclohexanol conformational switch. pH-sensitive fliposomes containing one or both of these flipids, as well as POPC and PEG ceramide, were constructed and characterized. These compositions were stable at 4°C and pH 7.4 for several months. Fliposomes loaded with ANTS/DPX performed an unusually quick content release within a few seconds at pH below 8.5 (in case of 2) and 6.0 (in case of 1). This difference in pH sensitivity demonstrates a potential for the custom design of flipids by variation of the amino group to target areas with specific pH values. The pH titration curves for the fliposome leakage parallel the curves for the acid-induced conformational flip of 1 and 2 studied by ¹H NMR. A plausible mechanism of pH sensitivity starts with an acid-triggered conformational flip of 1 or 2, which changes the molecular size and shape, shortens the lipid tails, and perturbs the liposome membrane, resulting in the content leakage.

  3. Extracellular Polymeric Substances from Shewanella sp. HRCR-1 Biofilms: Characterization by Infrared Spectroscopy and Proteomics

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cao, Bin; Shi, Liang; Brown, Roslyn N.

    This study characterizes the composition of extracellular polymeric substances (EPS) from Shewanella sp. HRCR-1 biofilms to provide insight into potential interactions of EPS with redox-active metals and radionuclides. Both bound and loosely associated EPS were extracted from Shewanella sp. HRCR-1 biofilms prepared using a hollow-fiber membrane biofilm reactor (HfMBR). FTIR spectra revealed the presence of proteins, polysaccharides, nucleic acids, membrane lipids, and fatty acids in both bound and loosely associated EPS. Using a global proteomic approach, a total of 58 extracellular and outer membrane proteins were identified in the EPS. These included homologues of multiple S. oneidensis MR-1 proteins thatmore » potentially contribute to key physiological biofilm processes, such as biofilm-promoting protein BpfA, surface-associated serine protease, nucleotidases (CpdB and UshA), an extracellular lipase, and oligopeptidases (PtrB and a M13 family oligopeptidase lipoprotein). In addition, 20 redox proteins were found in extracted EPS. Among the detected redox proteins were the homologues of two S. oneidensis MR-1 c-type cytochromes, MtrC and OmcA, which have been implicated in extracellular electron transfer. Given their detection in the EPS of Shewanella sp. HRCR 1 biofilms, c-type cytochromes may contribute to the possible redox activity of the biofilm matrix and play important roles in extracellular electron transfer reactions.« less

  4. Constitutive activation of a plasma membrane H(+)-ATPase prevents abscisic acid-mediated stomatal closure.

    PubMed

    Merlot, Sylvain; Leonhardt, Nathalie; Fenzi, Francesca; Valon, Christiane; Costa, Miguel; Piette, Laurie; Vavasseur, Alain; Genty, Bernard; Boivin, Karine; Müller, Axel; Giraudat, Jérôme; Leung, Jeffrey

    2007-07-11

    Light activates proton (H(+))-ATPases in guard cells, to drive hyperpolarization of the plasma membrane to initiate stomatal opening, allowing diffusion of ambient CO(2) to photosynthetic tissues. Light to darkness transition, high CO(2) levels and the stress hormone abscisic acid (ABA) promote stomatal closing. The overall H(+)-ATPase activity is diminished by ABA treatments, but the significance of this phenomenon in relationship to stomatal closure is still debated. We report two dominant mutations in the OPEN STOMATA2 (OST2) locus of Arabidopsis that completely abolish stomatal response to ABA, but importantly, to a much lesser extent the responses to CO(2) and darkness. The OST2 gene encodes the major plasma membrane H(+)-ATPase AHA1, and both mutations cause constitutive activity of this pump, leading to necrotic lesions. H(+)-ATPases have been traditionally assumed to be general endpoints of all signaling pathways affecting membrane polarization and transport. Our results provide evidence that AHA1 is a distinct component of an ABA-directed signaling pathway, and that dynamic downregulation of this pump during drought is an essential step in membrane depolarization to initiate stomatal closure.

  5. Alternative to antibiotics against Pseudomonas aeruginosa: Effects of Glycyrrhiza glabra on membrane permeability and inhibition of efflux activity and biofilm formation in Pseudomonas aeruginosa and its in vitro time-kill activity.

    PubMed

    Chakotiya, Ankita Singh; Tanwar, Ankit; Narula, Alka; Sharma, Rakesh Kumar

    2016-09-01

    The multi-drug resistance offered by Pseudomonas aeruginosa to antibiotics can be attributed towards its propensity to develop biofilm, modification in cell membrane and to efflux antibacterial drugs. The present study explored the activity of Glycyrrhiza glabra and one of its pure compounds, glycyrrhizic acid against P. aeruginosa and their mechanism of action in terms of the effect on membrane permeability, efflux activity, and biofilm formation were determined. Minimum inhibitory concentrations were determined by using broth dilution technique. The minimum bactericidal concentrations were assessed on agar plate. The MIC of the extract and glycyrrhizic acid was found to be 200 and 100 μg ml(-1), respectively. The MBC was found to be 800 and 400 μg ml(-1) in the case of extract and glycyrrhizic acid, respectively. Time -dependent killing efficacy was also estimated. Flowcytometric analysis with staining methods was used to determine the effect of extract and glycyrrhizic acid at 2 × MIC on different physiological parameters and compared it with the standard (antibiotic). The growth of P. aeruginosa was significantly inhibited by extract and the pure compound. The herbal extract and the glycyrrhic acid were also found to effective in targeting the physiological parameters of the bacteria that involve cell membrane permeabilization, efflux activity, and biofilm formation. This study reports the antipseudomonal action of Glycyrrhiza glabra and one of its compound and provides insight into their mode of action. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Membrane fouling and performance evaluation of conventional membrane bioreactor (MBR), moving biofilm MBR and oxic/anoxic MBR.

    PubMed

    Khan, Sher Jamal; Ahmad, Aman; Nawaz, Muhammad Saqib; Hankins, Nicholas P

    2014-01-01

    In this study, three laboratory scale submerged membrane bioreactors (MBRs) comprising a conventional MBR (C-MBR), moving bed MBR (MB-MBR) and anoxic-oxic MBR (A/O-MBR) were continuously operated with synthesized domestic wastewater (chemical oxygen demand, COD = 500 mg/L) for 150 days under similar operational and environmental conditions. Kaldnes(®) plastic media with 20% dry volume was used as a biofilm carrier in the MB-MBR and A/O-MBR. The treatment performance and fouling propensity of the MBRs were evaluated. The effect of cake layer formation in all three MBRs was almost the same. However, pore blocking caused a major difference in the resultant water flux. The A/O-MBR showed the highest total nitrogen and phosphorus (PO4-P) removal efficiencies of 83.2 and 69.7%, respectively. Due to the high removal of nitrogen, fewer protein contents were found in the soluble and bound extracellular polymeric substances (EPS) of the A/O-MBR. Fouling trends of the MBRs showed 12, 14 and 20 days filtration cycles for C-MBR, MB-MBR and A/O-MBR, respectively. A 25% reduction of the soluble EPS and a 37% reduction of the bound EPS concentrations in A/O-MBR compared with C-MBR was a major contributing factor for fouling retardation and the enhanced filtration capacity of the A/O-MBR.

  7. In vitro biofilm model for studying tongue flora and malodour.

    PubMed

    Spencer, P; Greenman, J; McKenzie, C; Gafan, G; Spratt, D; Flanagan, A

    2007-10-01

    To develop a perfusion biofilm system to model tongue biofilm microflora and their physiological response to sulfur-containing substrates (S-substrates) in terms of volatile sulfide compound (VSC) production. Tongue-scrape inocula were used to establish in vitro perfusion biofilms which were examined in terms of ecological composition using culture-dependent and independent (PCR-DGGE) approaches. VSC-specific activity of cells was measured by a cell suspension assay, using a portable industrial sulfide monitor which was also used to monitor VSC production from biofilms in situ. Quasi steady states were achieved by 48 h and continued to 96 h. The mean (+/-SEM) growth rate for 72-h biofilms (n=4) was micro=0.014 h(-1) (+/-0.005 h(-1)). Comparison of biofilms, perfusate and original inoculum showed their ecological composition to be similar (Pearson coefficient>0.64). Perfusate and biofilm cells derived from the same condition (co-sampled) were equivalent with regard to VSC-specific activities which were up-regulated in the presence of S-substrates. The model maintained a stable tongue microcosm suitable for studying VSC production; biofilm growth in the presence of S-substrates up-regulated VSC activity. The method is apt for studying ecological and physiological aspects of oral biofilms and could be useful for screening inhibitory agents.

  8. Cranberry Flavonoids Modulate Cariogenic Properties of Mixed-Species Biofilm through Exopolysaccharides-Matrix Disruption.

    PubMed

    Kim, Dongyeop; Hwang, Geelsu; Liu, Yuan; Wang, Yifei; Singh, Ajay P; Vorsa, Nicholi; Koo, Hyun

    2015-01-01

    The exopolysaccharides (EPS) produced by Streptococcus mutans-derived glucosyltransferases (Gtfs) are essential virulence factors associated with the initiation of cariogenic biofilms. EPS forms the core of the biofilm matrix-scaffold, providing mechanical stability while facilitating the creation of localized acidic microenvironments. Cranberry flavonoids, such as A-type proanthocyanidins (PACs) and myricetin, have been shown to inhibit the activity of Gtfs and EPS-mediated bacterial adhesion without killing the organisms. Here, we investigated whether a combination of cranberry flavonoids disrupts EPS accumulation and S. mutans survival using a mixed-species biofilm model under cariogenic conditions. We also assessed the impact of cranberry flavonoids on mechanical stability and the in situ pH at the biofilm-apatite interface. Topical application of an optimized combination of PACs oligomers (100-300 μM) with myricetin (2 mM) twice daily was used to simulate treatment regimen experienced clinically. Treatments with cranberry flavonoids effectively reduced the insoluble EPS content (>80% reduction vs. vehicle-control; p<0.001), while hindering S. mutans outgrowth within mixed-species biofilms. As a result, the 3D architecture of cranberry-treated biofilms was severely compromised, showing a defective EPS-matrix and failure to develop microcolonies on the saliva-coated hydroxyapatite (sHA) surface. Furthermore, topical applications of cranberry flavonoids significantly weaken the mechanical stability of the biofilms; nearly 90% of the biofilm was removed from sHA surface after exposure to a shear stress of 0.449 N/m2 (vs. 36% removal in vehicle-treated biofilms). Importantly, in situ pH measurements in cranberry-treated biofilms showed significantly higher pH values (5.2 ± 0.1) at the biofilm-apatite interface vs. vehicle-treated biofilms (4.6 ± 0.1). Altogether, the data provide important insights on how cranberry flavonoids treatments modulate virulence

  9. Effect of water temperature on biofouling development in reverse osmosis membrane systems.

    PubMed

    Farhat, N M; Vrouwenvelder, J S; Van Loosdrecht, M C M; Bucs, Sz S; Staal, M

    2016-10-15

    Understanding the factors that determine the spatial and temporal biofilm development is a key to formulate effective control strategies in reverse osmosis membrane systems for desalination and wastewater reuse. In this study, biofilm development was investigated at different water temperatures (10, 20, and 30 °C) inside a membrane fouling simulator (MFS) flow cell. The MFS studies were done at the same crossflow velocity with the same type of membrane and spacer materials, and the same feed water type and nutrient concentration, differing only in water temperature. Spatially resolved biofilm parameters such as oxygen decrease rate, biovolume, biofilm spatial distribution, thickness and composition were measured using in-situ imaging techniques. Pressure drop (PD) increase in time was used as a benchmark as to when to stop the experiments. Biofilm measurements were performed daily, and experiments were stopped once the average PD increased to 40 mbar/cm. The results of the biofouling study showed that with increasing feed water temperature (i) the biofilm activity developed faster, (ii) the pressure drop increased faster, while (iii) the biofilm thickness decreased. At an average pressure drop increase of 40 mbar/cm over the MFS for the different feed water temperatures, different biofilm activities, structures, and quantities were found, indicating that diagnosis of biofouling of membranes operated at different or varying (seasonal) feed water temperatures may be challenging. Membrane installations with a high temperature feed water are more susceptible to biofouling than installations fed with low temperature feed water. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. [Effects of traditional Chinese medicine on oral bacteria biofilm].

    PubMed

    Zhao, Jin; Li, Ji-yao; Zhu, Bing; Zhou, Xue-dong

    2007-10-01

    To investigate the effects of compounds of Galla chinensis extract (GCE) and Nidus vespae extract-1 (WVE1) on oral bacteria biofilm structure and activity and to determine the possibility of caries prevention by the compounds. The morphology and activity of treated-oral bacterial biofilm and untreated-oral bacterial biofilm were observed by using fluorescence microscope in combination of idio-fluorochrome to label the died and living bacteria. The visible light semiquantitative method was used to measure biomass glucosyltransferase (GTF, A620) values and to determine the effects of active compounds of GCE and NVE1 on GTF of oral bacteria biofilm. The living bacteria in the untreated 24 h bacterial biofilm was dominant, and only a small number of died bacteria were found, the biofilm structure was regular and clear. GCE, GCE-B and NVE1 could inhibit the bacteria in the dental biofilm, which showed significant difference with the negative control. GCE and NVE1 could also inhibit GTF activity of 24 h bacterial biofilm in comparison with the negative control. The traditional Chinese medicine Galla chinensis and Nidus vespae could not only inhibit bacteria growth on oral bacterial biofilm, but also function by adjusting biofilm structure, composition and GTF activity of 24 h bacterial biofilm.

  11. Community genomic analysis of an extremely acidophilic sulfur-oxidizing biofilm

    PubMed Central

    Jones, Daniel S; Albrecht, Heidi L; Dawson, Katherine S; Schaperdoth, Irene; Freeman, Katherine H; Pi, Yundan; Pearson, Ann; Macalady, Jennifer L

    2012-01-01

    Highly acidic (pH 0–1) biofilms, known as ‘snottites', form on the walls and ceilings of hydrogen sulfide-rich caves. We investigated the population structure, physiology and biogeochemistry of these biofilms using metagenomics, rRNA methods and lipid geochemistry. Snottites from the Frasassi cave system (Italy) are dominated (>70% of cells) by Acidithiobacillus thiooxidans, with smaller populations including an archaeon in the uncultivated ‘G-plasma' clade of Thermoplasmatales (>15%) and a bacterium in the Acidimicrobiaceae family (>5%). Based on metagenomic evidence, the Acidithiobacillus population is autotrophic (ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), carboxysomes) and oxidizes sulfur by the sulfide–quinone reductase and sox pathways. No reads matching nitrogen fixation genes were detected in the metagenome, whereas multiple matches to nitrogen assimilation functions are present, consistent with geochemical evidence, that fixed nitrogen is available in the snottite environment to support autotrophic growth. Evidence for adaptations to extreme acidity include Acidithiobacillus sequences for cation transporters and hopanoid synthesis, and direct measurements of hopanoid membrane lipids. Based on combined metagenomic, molecular and geochemical evidence, we suggest that Acidithiobacillus is the snottite architect and main primary producer, and that snottite morphology and distributions in the cave environment are directly related to the supply of C, N and energy substrates from the cave atmosphere. PMID:21716305

  12. A small molecule norspermidine in combination with silver ion enhances dispersal and disinfection of multi-species wastewater biofilms.

    PubMed

    Wu, Yachuan; Quan, Xiangchun; Si, Xiurong; Wang, Xinrui

    2016-06-01

    Detrimental biofilms have become a great concern in many areas due to their strong resistance and insensitivity to traditional antimicrobial agents. Norspermidine is a potent small molecule for biofilm dispersal. In this study, silver ion, a conventional inorganic biocide, was combined with norspermidine and used for control and removal of multi-species biofilms formed by a mixed culture from wastewater treatment systems. Results showed that silver ion (0.01-1 mg/L) treatment alone failed to remove the existing wastewater biofilms. Norspermidine at the concentrations of 500-1000 μM was capable to disrupt and disperse the existing biofilms with a biofilm reduction of 21-34 % after 24-h exposure. The combined treatment with norspermidine (500 μM) and silver ion (0.01 mg/L) increased biofilm reduction to 48 % (24-h exposure). The combined treatment also enhanced biofilm disinfection ratio (82 %, 2-h exposure) by 2.0- and 2.6-folds compared to norspermidine (27 %) or silver ion (23 %) treatment alone, respectively. Confocal laser scanning microscopic (CLSM) observations found that norspermidine could disrupt biofilm matrix and promote biofilm dispersal via breaking down exopolysaccharides. The combined treatment increased the reduction in biofilm cell density and viability, possibly due to the damage of biofilm matrix, enhanced silver ion diffusion in biofilms, and increased biofilm sensitivity. These findings indicate that the combination of a small molecule norspermidine with a traditional biocide silver ion presents a novel strategy to remove and kill biofilms, which have a potential application in addressing wastewater biofilm-related issues.

  13. The Xylella fastidiosa PD1063 protein is secreted in association with outer membrane vesicles.

    PubMed

    Pierce, Brittany K; Voegel, Tanja; Kirkpatrick, Bruce C

    2014-01-01

    Xylella fastidiosa is a gram-negative, xylem-limited plant pathogenic bacterium that causes disease in a variety of economically important agricultural crops including Pierce's disease of grapevines. Xylella fastidiosa biofilms formed in the xylem vessels of plants play a key role in early colonization and pathogenicity by providing a protected niche and enhanced cell survival. Here we investigate the role of Xylella fastidiosa PD1063, the predicted ortholog of Xanthomonas oryzae pv. oryzae PXO_03968, which encodes an outer membrane protein. To assess the function of the Xylella fastidiosa ortholog, we created Xylella fastidiosa mutants deleted for PD1063 and then assessed biofilm formation, cell-cell aggregation and cell growth in vitro. We also assessed disease severity and pathogen titers in grapevines mechanically inoculated with the Xylella fastidiosa PD1063 mutant. We found a significant decrease in cell-cell aggregation among PD1063 mutants but no differences in cell growth, biofilm formation, disease severity or titers in planta. Based on the demonstration that Xanthomonas oryzae pv. oryzae PXO_03968 encodes an outer membrane protein, secreted in association with outer membrane vesicles, we predicted that PD1063 would also be secreted in a similar manner. Using anti-PD1063 antibodies, we found PD1063 in the supernatant and secreted in association with outer membrane vesicles. PD1063 purified from the supernatant, outer membrane fractions and outer membrane vesicles was 19.2 kD, corresponding to the predicted size of the processed protein. Our findings suggest Xylella fastidiosa PD1063 is not essential for development of Pierce's disease in Vitis vinifera grapevines although further research is required to determine the function of the PD1063 outer membrane protein in Xylella fastidiosa.

  14. Proteus mirabilis biofilm - qualitative and quantitative colorimetric methods-based evaluation.

    PubMed

    Kwiecinska-Piróg, Joanna; Bogiel, Tomasz; Skowron, Krzysztof; Wieckowska, Ewa; Gospodarek, Eugenia

    2014-01-01

    Proteus mirabilis strains ability to form biofilm is a current topic of a number of research worldwide. In this study the biofilm formation of P. mirabilis strains derived from urine of the catheterized and non-catheterized patients has been investigated. A total number of 39 P. mirabilis strains isolated from the urine samples of the patients of dr Antoni Jurasz University Hospital No. 1 in Bydgoszcz clinics between 2011 and 2012 was used. Biofilm formation was evaluated using two independent quantitative and qualitative methods with TTC (2,3,5-triphenyl-tetrazolium chloride) and CV (crystal violet) application. The obtained results confirmed biofilm formation by all the examined strains, except quantitative method with TTC, in which 7.7% of the strains did not have this ability. It was shown that P. mirabilis rods have the ability to form biofilm on the surfaces of both biomaterials applied, polystyrene and polyvinyl chloride (Nelaton catheters). The differences in ability to form biofilm observed between P. mirabilis strains derived from the urine of the catheterized and non-catheterized patients were not statistically significant.

  15. Spatiotemporal pH dynamics in concentration polarization near ion-selective membranes.

    PubMed

    Andersen, Mathias B; Rogers, David M; Mai, Junyu; Schudel, Benjamin; Hatch, Anson V; Rempe, Susan B; Mani, Ali

    2014-07-08

    We present a detailed analysis of the transient pH dynamics for a weak, buffered electrolyte subject to voltage-driven transport through an ion-selective membrane. We show that pH fronts emanate from the concentration polarization zone next to the membrane and that these propagating fronts change the pH in the system several units from its equilibrium value. The analysis is based on a 1D model using the unsteady Poisson-Nernst-Planck equations with nonequilibrium chemistry and without assumptions of electroneutrality or asymptotically thin electric double layers. Nonequilibrium chemical effects, especially for water splitting, are shown to be important for the dynamical and spatiotemporal evolution of the pH fronts. Nonetheless, the model also shows that at steady state the assumption of chemical equilibrium can still lead to good approximations of the global pH distribution. Moreover, our model shows that the transport of the hydronium ion in the extended space charge region is governed by a balance between electromigration and water self-ionization. On the basis of this observation, we present a simple model showing that the net flux of the hydronium ion is proportional to the length of the extended space charge region and the water self-ionization rate. To demonstrate these effects in practice, we have adopted the experiment of Mai et al. (Mai, J.; Miller, H.; Hatch, A. V. Spatiotemporal Mapping of Concentration Polarization Induced pH Changes at Nanoconstrictions. ACS Nano 2012, 6, 10206) as a model problem, and by including the full chemistry and transport, we show that the present model can capture the experimentally observed pH fronts. Our model can, among other things, be used to predict and engineer pH dynamics, which can be essential to the performance of membrane-based systems for biochemical separation and analysis.

  16. Transport of H2S and HS− across the human red blood cell membrane: rapid H2S diffusion and AE1-mediated Cl−/HS− exchange

    PubMed Central

    2013-01-01

    The rates of H2S and HS− transport across the human erythrocyte membrane were estimated by measuring rates of dissipation of pH gradients in media containing 250 μM H2S/HS−. Net acid efflux is caused by H2S/HS− acting analogously to CO2/HCO3− in the Jacobs-Stewart cycle. The steps are as follows: 1) H2S efflux through the lipid bilayer and/or a gas channel, 2) extracellular H2S deprotonation, 3) HS− influx in exchange for Cl−, catalyzed by the anion exchange protein AE1, and 4) intracellular HS− protonation. Net acid transport by the Cl−/HS−/H2S cycle is more efficient than by the Cl−/HCO3−/CO2 cycle because of the rapid H2S-HS− interconversion in cells and medium. The rates of acid transport were analyzed by solving the mass flow equations for the cycle to produce estimates of the HS− and H2S transport rates. The data indicate that HS− is a very good substrate for AE1; the Cl−/HS− exchange rate is about one-third as rapid as Cl−/HCO3− exchange. The H2S permeability coefficient must also be high (>10−2 cm/s, half time <0.003 s) to account for the pH equilibration data. The results imply that H2S and HS− enter erythrocytes very rapidly in the microcirculation of H2S-producing tissues, thereby acting as a sink for H2S and lowering the local extracellular concentration, and the fact that HS− is a substrate for a Cl−/HCO3− exchanger indicates that some effects of exogenous H2S/HS− may not result from a regulatory role of H2S but, rather, from net acid flux by H2S and HS− transport in a Jacobs-Stewart cycle. PMID:23864610

  17. Transport of H2S and HS(-) across the human red blood cell membrane: rapid H2S diffusion and AE1-mediated Cl(-)/HS(-) exchange.

    PubMed

    Jennings, Michael L

    2013-11-01

    The rates of H2S and HS(-) transport across the human erythrocyte membrane were estimated by measuring rates of dissipation of pH gradients in media containing 250 μM H2S/HS(-). Net acid efflux is caused by H2S/HS(-) acting analogously to CO2/HCO3(-) in the Jacobs-Stewart cycle. The steps are as follows: 1) H2S efflux through the lipid bilayer and/or a gas channel, 2) extracellular H2S deprotonation, 3) HS(-) influx in exchange for Cl(-), catalyzed by the anion exchange protein AE1, and 4) intracellular HS(-) protonation. Net acid transport by the Cl(-)/HS(-)/H2S cycle is more efficient than by the Cl(-)/HCO3(-)/CO2 cycle because of the rapid H2S-HS(-) interconversion in cells and medium. The rates of acid transport were analyzed by solving the mass flow equations for the cycle to produce estimates of the HS(-) and H2S transport rates. The data indicate that HS(-) is a very good substrate for AE1; the Cl(-)/HS(-) exchange rate is about one-third as rapid as Cl(-)/HCO3(-) exchange. The H2S permeability coefficient must also be high (>10(-2) cm/s, half time <0.003 s) to account for the pH equilibration data. The results imply that H2S and HS(-) enter erythrocytes very rapidly in the microcirculation of H2S-producing tissues, thereby acting as a sink for H2S and lowering the local extracellular concentration, and the fact that HS(-) is a substrate for a Cl(-)/HCO3(-) exchanger indicates that some effects of exogenous H2S/HS(-) may not result from a regulatory role of H2S but, rather, from net acid flux by H2S and HS(-) transport in a Jacobs-Stewart cycle.

  18. Direct N2H4/H2O2 Fuel Cells Powered by Nanoporous Gold Leaves

    PubMed Central

    Yan, Xiuling; Meng, Fanhui; Xie, Yun; Liu, Jianguo; Ding, Yi

    2012-01-01

    Dealloyed nanoporous gold leaves (NPGLs) are found to exhibit high electrocatalytic properties toward both hydrazine (N2H4) oxidation and hydrogen peroxide (H2O2) reduction. This observation allows the implementation of a direct hydrazine-hydrogen peroxide fuel cell (DHHPFC) based on these novel porous membrane catalysts. The effects of fuel and oxidizer flow rate, concentration and cell temperature on the performance of DHHPFC are systematically investigated. With a loading of ~0.1 mg cm−2 Au on each side, an open circuit voltage (OCV) of 1.2 V is obtained at 80°C with a maximum power density 195 mW cm−2, which is 22 times higher than that of commercial Pt/C electrocatalyst at the same noble metal loading. NPGLs thus hold great potential as effective and stable electrocatalysts for DHHPFCs. PMID:23230507

  19. Simulation of Escherichia coli Dynamics in Biofilms and Submerged Colonies with an Individual-Based Model Including Metabolic Network Information.

    PubMed

    Tack, Ignace L M M; Nimmegeers, Philippe; Akkermans, Simen; Hashem, Ihab; Van Impe, Jan F M

    2017-01-01

    Clustered microbial communities are omnipresent in the food industry, e.g., as colonies of microbial pathogens in/on food media or as biofilms on food processing surfaces. These clustered communities are often characterized by metabolic differentiation among their constituting cells as a result of heterogeneous environmental conditions in the cellular surroundings. This paper focuses on the role of metabolic differentiation due to oxygen gradients in the development of Escherichia coli cell communities, whereby low local oxygen concentrations lead to cellular secretion of weak acid products. For this reason, a metabolic model has been developed for the facultative anaerobe E. coli covering the range of aerobic, microaerobic, and anaerobic environmental conditions. This metabolic model is expressed as a multiparametric programming problem, in which the influence of low extracellular pH values and the presence of undissociated acid cell products in the environment has been taken into account. Furthermore, the developed metabolic model is incorporated in MICRODIMS, an in-house developed individual-based modeling framework to simulate microbial colony and biofilm dynamics. Two case studies have been elaborated using the MICRODIMS simulator: (i) biofilm growth on a substratum surface and (ii) submerged colony growth in a semi-solid mixed food product. In the first case study, the acidification of the biofilm environment and the emergence of typical biofilm morphologies have been observed, such as the mushroom-shaped structure of mature biofilms and the formation of cellular chains at the exterior surface of the biofilm. The simulations show that these morphological phenomena are respectively dependent on the initial affinity of pioneer cells for the substratum surface and the cell detachment process at the outer surface of the biofilm. In the second case study, a no-growth zone emerges in the colony center due to a local decline of the environmental pH. As a result

  20. A mechanism of growth inhibition by abscisic acid in germinating seeds of Arabidopsis thaliana based on inhibition of plasma membrane H+-ATPase and decreased cytosolic pH, K+, and anions.

    PubMed

    Planes, María D; Niñoles, Regina; Rubio, Lourdes; Bissoli, Gaetano; Bueso, Eduardo; García-Sánchez, María J; Alejandro, Santiago; Gonzalez-Guzmán, Miguel; Hedrich, Rainer; Rodriguez, Pedro L; Fernández, José A; Serrano, Ramón

    2015-02-01

    The stress hormone abscisic acid (ABA) induces expression of defence genes in many organs, modulates ion homeostasis and metabolism in guard cells, and inhibits germination and seedling growth. Concerning the latter effect, several mutants of Arabidopsis thaliana with improved capability for H(+) efflux (wat1-1D, overexpression of AKT1 and ost2-1D) are less sensitive to inhibition by ABA than the wild type. This suggested that ABA could inhibit H(+) efflux (H(+)-ATPase) and induce cytosolic acidification as a mechanism of growth inhibition. Measurements to test this hypothesis could not be done in germinating seeds and we used roots as the most convenient system. ABA inhibited the root plasma-membrane H(+)-ATPase measured in vitro (ATP hydrolysis by isolated vesicles) and in vivo (H(+) efflux from seedling roots). This inhibition involved the core ABA signalling elements: PYR/PYL/RCAR ABA receptors, ABA-inhibited protein phosphatases (HAB1), and ABA-activated protein kinases (SnRK2.2 and SnRK2.3). Electrophysiological measurements in root epidermal cells indicated that ABA, acting through the PYR/PYL/RCAR receptors, induced membrane hyperpolarization (due to K(+) efflux through the GORK channel) and cytosolic acidification. This acidification was not observed in the wat1-1D mutant. The mechanism of inhibition of the H(+)-ATPase by ABA and its effects on cytosolic pH and membrane potential in roots were different from those in guard cells. ABA did not affect the in vivo phosphorylation level of the known activating site (penultimate threonine) of H(+)-ATPase in roots, and SnRK2.2 phosphorylated in vitro the C-terminal regulatory domain of H(+)-ATPase while the guard-cell kinase SnRK2.6/OST1 did not. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  1. Phosphorus and Cu2+ removal by periphytic biofilm stimulated by upconversion phosphors doped with Pr3+-Li.

    PubMed

    Zhu, Yan; Zhang, Jianhong; Zhu, Ningyuan; Tang, Jun; Liu, Junzhuo; Sun, Pengfei; Wu, Yonghong; Wong, Po Keung

    2018-01-01

    Upconversion phosphors (UCPs) can convert visible light into luminescence, such as UV, which can regulate the growth of microbes. Based on these fundamentals, the community composition of periphytic biofilms stimulated by UCPs doped with Pr 3+ -Li + was proposed to augment the removal of phosphorus (P) and copper (Cu). Results showed that the biofilms with community composition optimized by UCPs doped with Pr 3+ -Li + had high P and Cu 2+ removal rates. This was partly due to overall bacterial and algal abundance and biomass increases. The synergistic actions of algal, bacterial biomass and carbon metabolic capacity in the Pr-Li stimulated biofilms facilitated the removal of P and Cu 2+ . The results show that the stimulation of periphytic biofilms by lanthanide-doped UCPs is a promising approach for augmenting P and Cu 2+ removal. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Isothermal microcalorimetry provides new insights into biofilm variability and dynamics.

    PubMed

    Astasov-Frauenhoffer, Monika; Braissant, Olivier; Hauser-Gerspach, Irmgard; Daniels, Alma U; Weiger, Roland; Waltimo, Tuomas

    2012-12-01

    The purpose of this study was to investigate a three-species in vitro biofilm with peri-implantitis-related bacteria for its variability and metabolic activity. Streptococcus sanguinis, Fusobacterium nucleatum, and Porphyromonas gingivalis were suspended in simulated body fluid containing 0.2% glucose to form biofilms on polished, protein-coated implant-grade titanium disks over 72 h using a flow chamber system. Thereafter, biofilm-coated disks were characterized by scanning electron microscopy and fluorescence in situ hybridization/confocal laser scanning microscopy. To assess metabolic activity within the biofilms, their heat flow was recorded for 480 h at 37 °C by IMC. The microscopic methods revealed that the total number of bacteria in the biofilms varied slightly among specimens (2.59 × 10(4)  ± 0.67 × 10(4)  cells mm(-2) ), whereas all three species were found constantly with unchanged proportions (S. sanguinis 41.3 ± 4.8%, F. nucleatum 17.7 ± 2.1%, and P. gingivalis 41.0 ± 4.9%). IMC revealed minor differences in time-to-peak heat flow (20.6 ± 4.5 h), a trend consistent with the small variation in bacterial species proportions as shown by microscopy. Peak heat flow (35.8 ± 42.6 μW), mean heat flow (13.1 ± 22.0 μW), and total heat over 480 h (23.5 ± 37.2 J) showed very high variation. These IMC results may be attributed to differences in the initial cell counts and relative proportions of the three species, their distribution and embedment in exopolysaccharide matrix on the test specimens. The present results provide new insights into variability and dynamics of biofilms on titanium disks, aspects that should be explored in future studies of dental surfaces. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  3. A 96-well-plate-based optical method for the quantitative and qualitative evaluation of Pseudomonas aeruginosa biofilm formation and its application to susceptibility testing.

    PubMed

    Müsken, Mathias; Di Fiore, Stefano; Römling, Ute; Häussler, Susanne

    2010-08-01

    A major reason for bacterial persistence during chronic infections is the survival of bacteria within biofilm structures, which protect cells from environmental stresses, host immune responses and antimicrobial therapy. Thus, there is concern that laboratory methods developed to measure the antibiotic susceptibility of planktonic bacteria may not be relevant to chronic biofilm infections, and it has been suggested that alternative methods should test antibiotic susceptibility within a biofilm. In this paper, we describe a fast and reliable protocol for using 96-well microtiter plates for the formation of Pseudomonas aeruginosa biofilms; the method is easily adaptable for antimicrobial susceptibility testing. This method is based on bacterial viability staining in combination with automated confocal laser scanning microscopy. The procedure simplifies qualitative and quantitative evaluation of biofilms and has proven to be effective for standardized determination of antibiotic efficiency on P. aeruginosa biofilms. The protocol can be performed within approximately 60 h.

  4. Living biointerfaces based on non-pathogenic bacteria support stem cell differentiation

    NASA Astrophysics Data System (ADS)

    Hay, Jake J.; Rodrigo-Navarro, Aleixandre; Hassi, Karoliina; Moulisova, Vladimira; Dalby, Matthew J.; Salmeron-Sanchez, Manuel

    2016-02-01

    Lactococcus lactis, a non-pathogenic bacteria, has been genetically engineered to express the III7-10 fragment of human fibronectin as a membrane protein. The engineered L. lactis is able to develop biofilms on different surfaces (such as glass and synthetic polymers) and serves as a long-term substrate for mammalian cell culture, specifically human mesenchymal stem cells (hMSC). This system constitutes a living interface between biomaterials and stem cells. The engineered biofilms remain stable and viable for up to 28 days while the expressed fibronectin fragment induces hMSC adhesion. We have optimised conditions to allow long-term mammalian cell culture, and found that the biofilm is functionally equivalent to a fibronectin-coated surface in terms of osteoblastic differentiation using bone morphogenetic protein 2 (BMP-2) added to the medium. This living bacteria interface holds promise as a dynamic substrate for stem cell differentiation that can be further engineered to express other biochemical cues to control hMSC differentiation.

  5. Enhanced Biotransformation of Fluoranthene by Intertidally Derived Cunninghamella elegans under Biofilm-Based and Niche-Mimicking Conditions

    PubMed Central

    Mitra, Sayani; Pramanik, Arnab; Banerjee, Srijoni; Haldar, Saubhik; Gachhui, Ratan

    2013-01-01

    The aims of the investigation were to ascertain if surface attachment of Cunninghamella elegans and niche intertidal conditions provided in a bioreactor influenced biotransformation of fluoranthene by C. elegans. A newly designed polymethylmethacrylate (PMMA) conico-cylindrical flask (CCF) holding eight equidistantly spaced rectangular strips mounted radially on a circular disc allowed comparison of fluoranthene biotransformation between CCFs with a hydrophobic surface (PMMA-CCF) and a hydrophilic glass surface (GS-CCF) and a 500-ml Erlenmeyer flask (EF). Fluoranthene biotransformation was higher by 22-fold, biofilm growth was higher by 3-fold, and cytochrome P450 gene expression was higher by 2.1-fold when C. elegans was cultivated with 2% inoculum as biofilm culture in PMMA-CCF compared to planktonic culture in EF. Biotransformation was enhanced by 7-fold with 10% inoculum. The temporal pattern of biofilm progression based on three-channel fluorescence detection by confocal laser scanning microscopy demonstrated well-developed, stable biofilm with greater colocalization of fluoranthene within extracellular polymeric substances and filaments of the biofilm grown on PMMA in contrast to a glass surface. A bioreactor with discs rotating at 2 revolutions per day affording 6-hourly emersion and immersion mimicked the niche intertidal habitat of C. elegans and supported biofilm formation and transformation of fluoranthene. The amount of transformed metabolite was 3.5-fold, biofilm growth was 3-fold, and cytochrome P450 gene expression was 1.9-fold higher in the process mimicking the intertidal conditions than in a submerged process without disc rotation. In the CCF and reactor, where biofilm formation was comparatively greater, higher concentration of exopolysaccharides allowed increased mobilization of fluoranthene within the biofilm with consequential higher gene expression leading to enhanced volumetric productivity. PMID:24038685

  6. Stabilization of the H,K-ATPase M5M6 membrane hairpin by K+ ions. Mechanistic significance for p2-type atpases.

    PubMed

    Gatto, C; Lutsenko, S; Shin, J M; Sachs, G; Kaplan, J H

    1999-05-14

    The integral membrane protein, the gastric H,K-ATPase, is an alpha-beta heterodimer, with 10 putative transmembrane segments in the alpha-subunit and one such segment in the beta-subunit. All transmembrane segments remain within the membrane domain following trypsinization of the intact gastric H,K-ATPase in the presence of K+ ions, identified as M1M2, M3M4, M5M6, and M7, M8, M9, and M10. Removal of K+ ions from this digested preparation results in the selective loss of the M5M6 hairpin from the membrane. The release of the M5M6 fragment is directed to the extracellular phase as evidenced by the accumulation of the released M5M6 hairpin inside the sealed inside out vesicles. The stabilization of the M5M6 hairpin in the membrane phase by the transported cation as well as loss to the aqueous phase in the absence of the transported cation has been previously observed for another P2-type ATPase, the Na, K-ATPase (Lutsenko, S., Anderko, R., and Kaplan, J. H. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 7936-7940). Thus, the effects of the counter-transported cation on retention of the M5M6 segment in the membrane as compared with the other membrane pairs may be a general feature of P2-ATPase ion pumps, reflecting a flexibility of this region that relates to the mechanism of transport.

  7. Species-independent attraction to biofilms through electrical signaling

    PubMed Central

    Humphries, Jacqueline; Xiong, Liyang; Liu, Jintao; Prindle, Arthur; Yuan, Fang; Arjes, Heidi A.; Tsimring, Lev; Süel, Gürol M.

    2017-01-01

    Summary Bacteria residing within biofilm communities can coordinate their behavior through cell-to-cell signaling. However, it remains unclear if these signals can also influence the behavior of distant cells that are not part of the community. Using a microfluidic approach, we find that potassium ion channel-mediated electrical signaling generated by a Bacillus subtilis biofilm can attract distant cells. Integration of experiments and mathematical modeling indicates that extracellular potassium emitted from the biofilm alters the membrane potential of distant cells, thereby directing their motility. This electrically-mediated attraction appears to be a generic mechanism that enables cross-species interactions, as Pseudomonas aeruginosa cells also become attracted to the electrical signal released by the B. subtilis biofilm. Cells within a biofilm community can thus not only coordinate their own behavior, but also influence the behavior of diverse bacteria at a distance through long-range electrical signaling. PMID:28086091

  8. Extrinsic Cation Selectivity of 2D Membranes

    PubMed Central

    2017-01-01

    From a systematic study of the concentration driven diffusion of positive and negative ions across porous 2D membranes of graphene and hexagonal boron nitride (h-BN), we prove their cation selectivity. Using the current–voltage characteristics of graphene and h-BN monolayers separating reservoirs of different salt concentrations, we calculate the reversal potential as a measure of selectivity. We tune the Debye screening length by exchanging the salt concentrations and demonstrate that negative surface charge gives rise to cation selectivity. Surprisingly, h-BN and graphene membranes show similar characteristics, strongly suggesting a common origin of selectivity in aqueous solvents. For the first time, we demonstrate that the cation flux can be increased by using ozone to create additional pores in graphene while maintaining excellent selectivity. We discuss opportunities to exploit our scalable method to use 2D membranes for applications including osmotic power conversion. PMID:28157333

  9. Succinic acid-producing biofilms of Actinobacillus succinogenes: reproducibility, stability and productivity.

    PubMed

    Maharaj, K; Bradfield, M F A; Nicol, W

    2014-09-01

    Continuous anaerobic fermentations were performed in a biofilm reactor packed with Poraver® beads. Dilution rates (D) varied between 0.054 and 0.72 h(-1), and D-glucose and CO2 gas were used as carbon substrates. Steady-state conditions were shown to be repeatable and independent of the operational history. Production stability was achieved over periods exceeding 80 h at values of D below 0.32 h(-1). In these situations, steady-state variation (expressed as fluctuations in NaOH neutralisation flow rates) exhibited a standard deviation of less than 5 % while no indication of biofilm deactivation was detected. The total biomass amount was found to be independent of the dilution rate with an average dry concentration of 23.8 ± 2.9 g L(-1) obtained for all runs. This suggests that the attachment area controls the extent of biofilm accumulation. Specific succinic acid (SA) productivities, based on the total biomass amount, exhibited a substantial decrease with decreasing D. An SA volumetric productivity of 10.8 g L(-1) h(-1) was obtained at D = 0.7 h(-1)-the highest value reported to date in Actinobacillus succinogenes fermentations. SA yields on glucose increased with decreasing D, with a yield of 0.90 ± 0.01 g g(-1) obtained at a D of 0.054 h(-1). Production of formic acid approached zero with decreasing D, while the succinic to acetic acid ratio increased with decreasing D, resulting in an increasing SA yield on glucose.

  10. Bacterial biofilm mechanical properties persist upon antibiotic treatment and survive cell death

    NASA Astrophysics Data System (ADS)

    Zrelli, K.; Galy, O.; Latour-Lambert, P.; Kirwan, L.; Ghigo, J. M.; Beloin, C.; Henry, N.

    2013-12-01

    Bacteria living on surfaces form heterogeneous three-dimensional consortia known as biofilms, where they exhibit many specific properties one of which is an increased tolerance to antibiotics. Biofilms are maintained by a polymeric network and display physical properties similar to that of complex fluids. In this work, we address the question of the impact of antibiotic treatment on the physical properties of biofilms based on recently developed tools enabling the in situ mapping of biofilm local mechanical properties at the micron scale. This approach takes into account the material heterogeneity and reveals the spatial distribution of all the small changes that may occur in the structure. With an Escherichia coli biofilm, we demonstrate using in situ fluorescent labeling that the two antibiotics ofloxacin and ticarcillin—targeting DNA replication and membrane assembly, respectively—induced no detectable alteration of the biofilm mechanical properties while they killed the vast majority of the cells. In parallel, we show that a proteolytic enzyme that cleaves extracellular proteins into short peptides, but does not alter bacterial viability in the biofilm, clearly affects the mechanical properties of the biofilm structure, inducing a significant increase of the material compliance. We conclude that conventional biofilm control strategy relying on the use of biocides targeting cells is missing a key target since biofilm structural integrity is preserved. This is expected to efficiently promote biofilm resilience, especially in the presence of persister cells. In contrast, the targeting of polymer network cross-links—among which extracellular proteins emerge as major players—offers a promising route for the development of rational multi-target strategies to fight against biofilms.

  11. Comparative analysis of soybean plasma membrane proteins under osmotic stress using gel-based and LC MS/MS-based proteomics approaches.

    PubMed

    Nouri, Mohammad-Zaman; Komatsu, Setsuko

    2010-05-01

    To study the soybean plasma membrane proteome under osmotic stress, two methods were used: a gel-based and a LC MS/MS-based proteomics method. Two-day-old seedlings were subjected to 10% PEG for 2 days. Plasma membranes were purified from seedlings using a two-phase partitioning method and their purity was verified by measuring ATPase activity. Using the gel-based proteomics, four and eight protein spots were identified as up- and downregulated, respectively, whereas in the nanoLC MS/MS approach, 11 and 75 proteins were identified as up- and downregulated, respectively, under PEG treatment. Out of osmotic stress responsive proteins, most of the transporter proteins and all proteins with high number of transmembrane helices as well as low-abundance proteins could be identified by the LC MS/MS-based method. Three homologues of plasma membrane H(+)-ATPase, which are transporter proteins involved in ion efflux, were upregulated under osmotic stress. Gene expression of this protein was increased after 12 h of stress exposure. Among the identified proteins, seven proteins were mutual in two proteomics techniques, in which calnexin was the highly upregulated protein. Accumulation of calnexin in plasma membrane was confirmed by immunoblot analysis. These results suggest that under hyperosmotic conditions, calnexin accumulates in the plasma membrane and ion efflux accelerates by upregulation of plasma membrane H(+)-ATPase protein.

  12. Cyanex based uranyl sensitive polymeric membrane electrodes.

    PubMed

    Badr, Ibrahim H A; Zidan, W I; Akl, Z F

    2014-01-01

    Novel uranyl selective polymeric membrane electrodes were prepared using three different low-cost and commercially available Cyanex extractants namely, bis(2,4,4-trimethylpentyl) phosphinic acid [L1], bis(2,4,4-trimethylpentyl) monothiophosphinic acid [L2] and bis(2,4,4-trimethylpentyl) dithiophosphinic acid [L3]. Optimization and performance characteristics of the developed Cyanex based polymer membrane electrodes were determined. The influence of membrane composition (e.g., amount and type of ionic sites, as well as type of plasticizer) on potentiometric responses of the prepared membrane electrodes was studied. Optimized Cyanex-based membrane electrodes exhibited Nernstian responses for UO₂(2+) ion over wide concentration ranges with fast response times. The optimized membrane electrodes based on L1, L2 and L3 exhibited Nernstian responses towards uranyl ion with slopes of 29.4, 28.0 and 29.3 mV decade(-1), respectively. The optimized membrane electrodes based on L1-L3 showed detection limits of 8.3 × 10(-5), 3.0 × 10(-5) and 3.3 × 10(-6) mol L(-1), respectively. The selectivity studies showed that the optimized membrane electrodes exhibited high selectivity towards UO₂(2+) ion over large number of other cations. Membrane electrodes based on L3 exhibited superior potentiometric response characteristics compared to those based on L1 and L2 (e.g., widest linear range and lowest detection limit). The analytical utility of uranyl membrane electrodes formulated with Cyanex extractant L3 was demonstrated by the analysis of uranyl ion in different real samples for nuclear safeguards verification purposes. The results obtained using direct potentiometry and flow-injection methods were compared with those measured using the standard UV-visible and inductively coupled plasma spectroscopic methods. © 2013 Published by Elsevier B.V.

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

    PubMed

    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.

  14. In vitro biofilm formation on resin-based composites after different finishing and polishing procedures.

    PubMed

    Cazzaniga, Gloria; Ottobelli, Marco; Ionescu, Andrei C; Paolone, Gaetano; Gherlone, Enrico; Ferracane, Jack L; Brambilla, Eugenio

    2017-12-01

    To evaluate the influence of surface treatments of different resin-based composites (RBCs) on S. mutans biofilm formation. 4 RBCs (microhybrid, nanohybrid, nanofilled, bulk-filled) and 6 finishing-polishing (F/P) procedures (open-air light-curing, light-curing against Mylar strip, aluminum oxide discs, one-step rubber point, diamond bur, multi-blade carbide bur) were evaluated. Surface roughness (SR) (n=5/group), gloss (n=5/group), scanning electron microscopy morphological analysis (SEM), energy-dispersive X-ray spectrometry (EDS) (n=3/group), and S. mutans biofilm formation (n=16/group) were assessed. EDS analysis was repeated after the biofilm assay. A morphological evaluation of S. mutans biofilm was also performed using confocal laser-scanning microscopy (CLSM) (n=2/group). The data were analyzed using Wilcoxon (SR, gloss) and two-way ANOVA with Tukey as post-hoc tests (EDS, biofilm formation). F/P procedures as well as RBCs significantly influenced SR and gloss. While F/P procedures did not significantly influence S. mutans biofilm formation, a significant influence of RBCs on the same parameter was found. Different RBCs showed different surface elemental composition. Both F/P procedures and S. mutans biofilm formation significantly modified this parameter. The tested F/P procedures significantly influenced RBCs surface properties but did not significantly affect S. mutans biofilm formation. The significant influence of the different RBCs tested on S. mutans biofilm formation suggests that material characteristics and composition play a greater role than SR. F/P procedures of RBCs may unexpectedly play a minor role compared to that of the restoration material itself in bacterial colonization. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Comparison of genes required for H2O2 resistance in Streptococcus gordonii and Streptococcus sanguinis

    PubMed Central

    Xu, Yifan; Itzek, Andreas

    2014-01-01

    Hydrogen peroxide (H2O2) is produced by several members of the genus Streptococcus mainly through the pyruvate oxidase SpxB under aerobic growth conditions. The acute toxic nature of H2O2 raises the interesting question of how streptococci cope with intrinsically produced H2O2, which subsequently accumulates in the microenvironment and threatens the closely surrounding population. Here, we investigate the H2O2 susceptibility of oral Streptococcus gordonii and Streptococcus sanguinis and elucidate potential mechanisms of how they protect themselves from the deleterious effect of H2O2. Both organisms are considered primary colonizers and occupy the same intraoral niche making them potential targets for H2O2 produced by other species. We demonstrate that S. gordonii produces relatively more H2O2 and has a greater ability for resistance to H2O2 stress. Functional studies show that, unlike in Streptococcus pneumoniae, H2O2 resistance is not dependent on a functional SpxB and confirms the important role of the ferritin-like DNA-binding protein Dps. However, the observed increased H2O2 resistance of S. gordonii over S. sanguinis is likely to be caused by an oxidative stress protection machinery present even under anaerobic conditions, while S. sanguinis requires a longer period of time for adaptation. The ability to produce more H2O2 and be more resistant to H2O2 might aid S. gordonii in the competitive oral biofilm environment, since it is lower in abundance yet manages to survive quite efficiently in the oral biofilm. PMID:25280752

  16. Extracellular Polymeric Substances (EPS) of Freshwater Biofilms Stabilize and Modify CeO2 and Ag Nanoparticles

    PubMed Central

    Kroll, Alexandra; Behra, Renata; Kaegi, Ralf; Sigg, Laura

    2014-01-01

    Streams are potential receiving compartments for engineered nanoparticles (NP). In streams, NP may remain dispersed or settle to the benthic compartment. Both dispersed and settling NP can accumulate in benthic biofilms called periphyton that are essential to stream ecosystems. Periphytic organisms excrete extracellular polymeric substances (EPS) that interact with any material reaching the biofilms. To understand the interaction of NP with periphyton it is therefore crucial to study the interaction of NP with EPS. We investigated the influence of EPS on the physicochemical properties of selected NP (CeO2, Ag) under controlled conditions at pH 6, 7.6, 8.6 and light or dark exposure. We extracted EPS from five different periphyton communities, characterized the extracts, and exposed CeO2 and carbonate-stabilized Ag NP (0.5 and 5 mg/L, both 25 nm primary particle size) and AgNO3 to EPS (10 mg/L) over two weeks. We measured NP size distribution, shape, primary particle size, surface plasmon resonance, and dissolution. All EPS extracts were composed of biopolymers, building blocks of humic substances, low molecular weight (Mr) acids, and small amphiphilic or neutral compounds in varying concentrations. CeO2 NP were stabilized by EPS independent of pH and light/dark while dissolution increased over time in the dark at pH 6. EPS induced a size increase in Ag NP in the light with decreasing pH and the formation of metallic Ag NP from AgNO3 at the same conditions via EPS-enhanced photoreduction. NP transformation and formation were slower in the extract with the lowest biopolymer and low Mr acid concentrations. Periphytic EPS in combination with naturally varying pH and light/dark conditions influence the properties of the Ag and CeO2 NP tested and thus the exposure conditions within biofilms. Our results indicate that periphytic organisms may be exposed to a constantly changing mixture of engineered and naturally formed Ag NP and Ag+. PMID:25333364

  17. Gas separation performance of carbon molecular sieve membranes based on 6FDA-mPDA/DABA (3:2) polyimide.

    PubMed

    Qiu, Wulin; Zhang, Kuang; Li, Fuyue Stephanie; Zhang, Ke; Koros, William J

    2014-04-01

    6FDA-mPDA/DABA (3:2) polyimide was synthesized and characterized for uncross-linked, thermally crosslinked, and carbon molecular sieve (CMS) membranes. The membranes were characterized with thermogravimetric analysis, FTIR spectroscopy, wide-angle X-ray diffraction, and gas permeation tests. Variations in the d spacing, the formation of pore structures, and changes in the pore sizes of the CMS membranes were discussed in relation to pyrolysis protocols. The uncross-linked polymer membranes showed high CO2 /CH4 selectivity, whereas thermally crosslinked membranes exhibited significantly improved CO2 permeability and excellent CO2 plasticization resistance. The CMS membranes showed even higher CO2 permeability and CO2 /CH4 selectivity. An increase in the pyrolysis temperature resulted in CMS membranes with lower gas permeability but higher selectivity. The 550 °C pyrolyzed CMS membranes showed CO2 permeability as high as 14 750 Barrer with CO2 /CH4 selectivity of approximately 52. Even 800 °C pyrolyzed CMS membranes still showed high CO2 permeability of 2610 Barrer with high CO2 /CH4 selectivity of approximately 118. Both polymer membranes and the CMS membranes are very attractive in aggressive natural gas purification applications. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Bactericidal effect of photocatalytically-active nanostructured TiO2 surfaces on biofilms of the early oral colonizer, Streptococcus oralis.

    PubMed

    Westas, Emma; Hayashi, Mariko; Cecchinato, Francesca; Wennerberg, Ann; Andersson, Martin; Jimbo, Ryo; Davies, Julia R

    2017-08-01

    This study evaluated the photocatalytic bactericidal effect of nanostructured anatase-rich titanium dioxide (TiO 2 ) on microbial biofilms. Commercially pure titanium discs were spin-coated with photocatalytic TiO 2 nanoparticles (P25). Uncoated discs were used as control (CTRL). Half of the CTRL and half of the P25-coated surfaces were coated with purified saliva (SAL) to give four different groups (CTRL, CTRL + SAL, P25 and P25 + SAL). Streptococcus oralis were allowed to form biofilms on the discs for 18 h and non-adherent cells were rinsed off. Bacterial viability was assessed at time 0 with Live/Dead BacLight staining and epifluorescence microscopy. The remaining discs were divided into a non-UV group and UVA-irradiated (+UV) group (irradiation time, 6 or 24 h). Thereafter, viability was assessed as above. Viability at time 0 was high and no dead cells were seen on any of the surfaces, even after 24 h, in the absence of UVA. However, after 24 h of exposure, the proportion of viable cells was reduced by 40% on the P25 discs compared to 0 and 6 h, and this effect was enhanced with a salivary pellicle. Members of mixed species biofilms differ in their susceptibility to the bactericidal effect of the surfaces tested and further investigations are needed to optimize the conditions. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2321-2328, 2017. © 2017 Wiley Periodicals, Inc.

  19. Biofilm Formation Characteristics of Pseudomonas lundensis Isolated from Meat.

    PubMed

    Liu, Yong-Ji; Xie, Jing; Zhao, Li-Jun; Qian, Yun-Fang; Zhao, Yong; Liu, Xiao

    2015-12-01

    Biofilms formations of spoilage and pathogenic bacteria on food or food contact surfaces have attracted increasing attention. These events may lead to a higher risk of food spoilage and foodborne disease transmission. While Pseudomonas lundensis is one of the most important bacteria that cause spoilage in chilled meat, its capability for biofilm formation has been seldom reported. Here, we investigated biofilm formation characteristics of P. lundensis mainly by using crystal violet staining, and confocal laser scanning microscopy (CLSM). The swarming and swimming motility, biofilm formation in different temperatures (30, 10, and 4 °C) and the protease activity of the target strain were also assessed. The results showed that P. lundensis showed a typical surface-associated motility and was quite capable of forming biofilms in different temperatures (30, 10, and 4 °C). The strain began to adhere to the contact surfaces and form biofilms early in the 4 to 6 h. The biofilms began to be formed in massive amounts after 12 h at 30 °C, and the extracellular polysaccharides increased as the biofilm structure developed. Compared with at 30 °C, more biofilms were formed at 4 and 10 °C even by a low bacterial density. The protease activity in the biofilm was significantly correlated with the biofilm formation. Moreover, the protease activity in biofilm was significantly higher than that of the corresponding planktonic cultures after cultured 12 h at 30 °C. © 2015 Institute of Food Technologists®

  20. Inhibition factors and Kinetic model for ammonium inhibition on the anammox process of the SNAD biofilm.

    PubMed

    Zheng, Zhaoming; Li, Jun; Ma, Jing; Du, Jia; Wang, Fan; Bian, Wei; Zhang, Yanzhuo; Zhao, Baihang

    2017-03-01

    The aim of the present work was to evaluate the anaerobic ammonium oxidation (anammox) activity of simultaneous partial nitrification, anammox and denitrification (SNAD) biofilm with different substrate concentrations and pH values. Kaldnes rings taken from the SNAD biofilm reactor were incubated in batch tests to determine the anammox activity. Haldane model was applied to investigate the ammonium inhibition on anammox process. As for nitrite inhibition, the NH 4 + -N removal rate of anammox process remained 87.4% of the maximum rate with the NO 2 - -N concentration of 100mg/L. Based on the results of Haldane model, no obvious difference in kinetic coefficients was observed under high or low free ammonia (FA) conditions, indicating that ammonium rather than FA was the true inhibitor for anammox process of SNAD biofilm. With the pH value of 7.0, the r max , Ks and K I of ammonium were 0.209kg NO 2 - -N/kg VSS/day, 9.5mg/L and 422mg/L, respectively. The suitable pH ranges for anammox process were 5.0 to 9.0. These results indicate that the SNAD biofilm performs excellent tolerance to adverse conditions. Copyright © 2016. Published by Elsevier B.V.

  1. The biofilm matrix destabilizers, EDTA and DNaseI, enhance the susceptibility of nontypeable Hemophilus influenzae biofilms to treatment with ampicillin and ciprofloxacin

    PubMed Central

    Cavaliere, Rosalia; Ball, Jessica L; Turnbull, Lynne; Whitchurch, Cynthia B

    2014-01-01

    Nontypeable Hemophilus influenzae (NTHi) is a Gram-negative bacterial pathogen that causes chronic biofilm infections of the ears and airways. The biofilm matrix provides structural integrity to the biofilm and protects biofilm cells from antibiotic exposure by reducing penetration of antimicrobial compounds into the biofilm. Extracellular DNA (eDNA) has been found to be a major matrix component of biofilms formed by many species of Gram-positive and Gram-negative bacteria, including NTHi. Interestingly, the cation chelator ethylenediaminetetra-acetic acid (EDTA) has been shown to reduce the matrix strength of biofilms of several bacterial species as well as to have bactericidal activity against various pathogens. EDTA exerts its antimicrobial activity by chelating divalent cations necessary for growth and membrane stability and by destabilizing the matrix thus enhancing the detachment of bacterial cells from the biofilm. In this study, we have explored the role of divalent cations in NTHi biofilm development and stability. We have utilized in vitro static and continuous flow models of biofilm development by NTHi to demonstrate that magnesium cations enhance biofilm formation by NTHi. We found that the divalent cation chelator EDTA is effective at both preventing NTHi biofilm formation and at treating established NTHi biofilms. Furthermore, we found that the matrix destablilizers EDTA and DNaseI increase the susceptibility of NTHi biofilms to ampicillin and ciprofloxacin. Our observations indicate that DNaseI and EDTA enhance the efficacy of antibiotic treatment of NTHi biofilms. These observations may lead to new strategies that will improve the treatment options available to patients with chronic NTHi infections. PMID:25044339

  2. The antibacterial activity of chlorhexidine digluconate against Streptococcus mutans biofilms follows sigmoidal patterns.

    PubMed

    Lee, Dae-Woo; Jung, Ji-Eun; Yang, Yeon-Mi; Kim, Jae-Gon; Yi, Ho-Keun; Jeon, Jae-Gyu

    2016-10-01

    The aim of this study was to determine the pattern of the antibacterial activity of chlorhexidine digluconate (CHX) against mature Streptococcus mutans biofilms. Streptococcus mutans biofilms were formed on saliva-coated hydroxyapatite discs and then treated with 0-20% CHX, once, three times, or five times (1 min per treatment) during the period of mature biofilm formation (beyond 46 h). After the treatments, the colony-forming unit (CFU) counts of the treated biofilms were determined. The pH values of the spent culture medium were also determined to investigate the change in pH resulting from the antibacterial activity of CHX. The relationships between the concentration of CHX and the CFU counts and the concentration of CHX and culture medium pH, relative to the number of treatments performed, were evaluated using a sigmoidal curve-fitting procedure. The changes in CFU counts and culture medium pH followed sigmoidal curves and were dependent on the concentration of CHX (R 2 = 0.99). The sigmoidal curves were left-shifted with increasing number of treatments. Furthermore, the culture-medium pH of the treated biofilms increased as their CFU counts decreased. The lowest CHX concentration to increase culture-medium pH above the critical pH also decreased as the number of treatments increased. These results may provide fundamental information for selecting the appropriate CHX concentrations to treat S. mutans biofilms. © 2016 Eur J Oral Sci.

  3. Mycobacterium avium Genes Associated with the Ability To Form a Biofilm

    PubMed Central

    Yamazaki, Yoshitaka; Danelishvili, Lia; Wu, Martin; MacNab, Molly; Bermudez, Luiz E.

    2006-01-01

    Mycobacterium avium is widely distributed in the environment, and it is chiefly found in water and soil. M. avium, as well as Mycobacterium smegmatis, has been recognized to produce a biofilm or biofilm-like structure. We screened an M. avium green fluorescent protein (GFP) promoter library in M. smegmatis for genes involved in biofilm formation on polyvinyl chloride (PVC) plates. Clones associated with increased GFP expression ≥2.0-fold over the baseline were sequenced. Seventeen genes, most encoding proteins of the tricarboxylic acid (TCA) cycle and GDP-mannose and fatty acid biosynthesis, were identified. Their regulation in M. avium was confirmed by examining the expression of a set of genes by real-time PCR after incubation on PVC plates. In addition, screening of 2,000 clones of a transposon mutant bank constructed using M. avium strain A5, a mycobacterial strain with the ability to produce large amounts of biofilm, revealed four mutants with an impaired ability to form biofilm. Genes interrupted by transposons were homologues of M. tuberculosis 6-oxodehydrogenase (sucA), enzymes of the TCA cycle, protein synthetase (pstB), enzymes of glycopeptidolipid (GPL) synthesis, and Rv1565c (a hypothetical membrane protein). In conclusion, it appears that GPL biosynthesis, including the GDP-mannose biosynthesis pathway, is the most important pathway involved in the production of M. avium biofilm. PMID:16391123

  4. Antimicrobial and Antibiofilm Effects of Human Amniotic/Chorionic Membrane Extract on Streptococcus pneumoniae

    PubMed Central

    Yadav, Mukesh K.; Go, Yoon Y.; Kim, Shin Hye; Chae, Sung-Won; Song, Jae-Jun

    2017-01-01

    Background: Streptococcus pneumoniae colonize the human nasopharynx in the form of biofilms. The biofilms act as bacterial reservoirs and planktonic bacteria from these biofilms can migrate to other sterile anatomical sites to cause pneumonia, otitis media (OM), bacteremia and meningitis. Human amniotic membrane contains numerous growth factors and antimicrobial activity; however, these have not been studied in detail. In this study, we prepared amniotic membrane extract and chorionic membrane extract (AME/CME) and evaluated their antibacterial and antibiofilm activities against S. pneumoniae using an in vitro biofilm model and in vivo OM rat model. Materials and Methods: The AME/CME were prepared and protein was quantified using DCTM (detergent compatible) method. The minimum inhibitory concentrations were determined using broth dilution method, and the synergistic effect of AME/CME with Penicillin-streptomycin was detected checkerboard. The in vitro biofilm and in vivo colonization of S. pneumoniae were studied using microtiter plate assay and OM rat model, respectively. The AME/CME-treated biofilms were examined using scanning electron microscope and confocal microscopy. To examine the constituents of AME/CME, we determined the proteins and peptides of AME/CME using tandem mass tag-based quantitative mass spectrometry. Results: AME/CME treatment significantly (p < 0.05) inhibited S. pneumoniae growth in planktonic form and in biofilms. Combined application of AME/CME and Penicillin-streptomycin solution had a synergistic effect against S. pneumoniae. Biofilms grown with AME/CME were thin, scattered, and unorganized. AME/CME effectively eradicated pre-established pneumococci biofilms and has a bactericidal effect. AME treatment significantly (p < 0.05) reduced bacterial colonization in the rat middle ear. The proteomics analysis revealed that the AME/CME contains hydrolase, ribonuclease, protease, and other antimicrobial proteins and peptides. Conclusion: AME

  5. Antibiotic-loaded MoS2 nanosheets to combat bacterial resistance via biofilm inhibition

    NASA Astrophysics Data System (ADS)

    Zhang, Xu; Zhang, Wentao; Liu, Lizhi; Yang, Mei; Huang, Lunjie; Chen, Kai; Wang, Rong; Yang, Baowei; Zhang, Daohong; Wang, Jianlong

    2017-06-01

    The emergence of antibiotic resistance has resulted in increasing difficulty in treating clinical infections associated with biofilm formation, one of the key processes in turn contributing to enhanced antibiotic resistance. With the rapid development of nanotechnology, a new way to overcome antibiotic resistance has opened up. Based on the many and diverse properties of MoS2 nanosheets that have attracted wide attention, in particular their antibacterial potential, herein, a novel antimicrobial agent to combat resistant gram-positive Staphylococcus aureus and gram-negative Salmonella was prepared using chitosan functionalized MoS2 nanosheets loading tetracycline hydrochloride drugs (abbreviated to CM-TH). The antibacterial and anti-biofilm activities of the CM-TH nanocomposites showed the synergetic effect that the combination of nanomaterials and antibiotics was more efficient than either working alone. In particularly, the minimum inhibitory concentration values generally decreased by a factor of dozens, suggesting that CM-TH may become a possible alternative to traditional antibiotics in disrupting biofilms and overcoming antibiotic resistance in treating medical diseases.

  6. Evolution of resistive switching mechanism through H2O2 sensing by using TaOx-based material in W/Al2O3/TaOx/TiN structure

    NASA Astrophysics Data System (ADS)

    Chakrabarti, Somsubhra; Panja, Rajeswar; Roy, Sourav; Roy, Anisha; Samanta, Subhranu; Dutta, Mrinmoy; Ginnaram, Sreekanth; Maikap, Siddheswar; Cheng, Hsin-Ming; Tsai, Ling-Na; Chang, Ya-Ling; Mahapatra, Rajat; Jana, Debanjan; Qiu, Jian-Tai; Yang, Jer-Ren

    2018-03-01

    Understanding of resistive switching mechanism through H2O2 sensing and improvement of switching characteristics by using TaOx-based material in W/Al2O3/TaOx/TiN structure have been reported for the first time. Existence of amorphous Al2O3/TaOx layer in the RRAM devices has been confirmed by transmission electron microscopy. By analyzing the oxidation states of Ta2+/Ta5+ for TaOx switching material and W0/W6+ for WOx layer at the W/TaOx interface through X-ray photoelectron spectroscopy and H2O2 sensing, the reduction-oxidation mechanism under Set/Reset occurs only in the TaOx layer for the W/Al2O3/TaOx/TiN structures. This leads to higher Schottky barrier height at the W/Al2O3 interface (0.54 eV vs. 0.46 eV), higher resistance ratio, and long program/erase endurance of >108 cycles with 100 ns pulse width at a low operation current of 30 μA. Stable retention of more than 104 s at 85 °C is also obtained. Using conduction mechanism and reduction-oxidation reaction, current-voltage characteristic has been simulated. Both TaOx and WOx membranes have high pH sensitivity values of 47.65 mV/pH and 49.25 mV/pH, respectively. Those membranes can also sense H2O2 with a low concentration of 1 nM in an electrolyte-insulator-semiconductor structure because of catalytic activity, while the Al2O3 membrane does not show sensing. The TaOx material in W/Al2O3/TaOx/TiN structure does not show only a path towards high dense, small size memory application with understanding of switching mechanism but also can be used for H2O2 sensors.

  7. Helicobacter pylori Resists the Antimicrobial Activity of Calprotectin via Lipid A Modification and Associated Biofilm Formation

    PubMed Central

    Gaddy, Jennifer A.; Radin, Jana N.; Cullen, Thomas W.; Chazin, Walter J.; Skaar, Eric P.; Trent, M. Stephen

    2015-01-01

    ABSTRACT Helicobacter pylori is one of several pathogens that persist within the host despite a robust immune response. H. pylori elicits a proinflammatory response from host epithelia, resulting in the recruitment of immune cells which manifests as gastritis. Relatively little is known about how H. pylori survives antimicrobials, including calprotectin (CP), which is present during the inflammatory response. The data presented here suggest that one way H. pylori survives the nutrient sequestration by CP is through alteration of its outer membrane. CP-treated H. pylori demonstrates increased bacterial fitness in response to further coculture with CP. Moreover, CP-treated H. pylori cultures form biofilms and demonstrate decreased cell surface hydrophobicity. In response to CP, the H. pylori Lpx lipid A biosynthetic enzymes are not fully functional. The lipid A molecules observed in H. pylori cultures treated with CP indicate that the LpxF, LpxL, and LpxR enzyme functions are perturbed. Transcriptional analysis of lpxF, lpxL, and lpxR indicates that metal restriction by CP does not control this pathway through transcriptional regulation. Analyses of H. pylori lpx mutants reveal that loss of LpxF and LpxL results in increased fitness, similar to what is observed in the presence of CP; moreover, these mutants have significantly increased biofilm formation and reduced cell surface hydrophobicity. Taken together, these results demonstrate a novel mechanism of H. pylori resistance to the antimicrobial activity of CP via lipid A modification strategies and resulting biofilm formation. PMID:26646009

  8. Autohydrogenotrophic denitrification of drinking water using a polyvinyl chloride hollow fiber membrane biofilm reactor.

    PubMed

    Zhang, Yanhao; Zhong, Fohua; Xia, Siqing; Wang, Xuejiang; Li, Jixiang

    2009-10-15

    A hollow fiber membrane biofilm reactor (MBfR) using polyvinyl chloride (PVC) hollow fiber was evaluated in removing nitrate form contaminated drinking water. During a 279-day operation period, the denitrification rate increased gradually with the increase of influent nitrate loading. The denitrification rate reached a maximum value of 414.72 g N/m(3)d (1.50 g N/m(2)d) at an influent NO(3)(-)-N concentration of 10mg/L and a hydraulic residence time of 37.5 min, and the influent nitrate was completely reduced. At the same time, the effluent quality analysis showed the headspace hydrogen content (3.0%) was lower enough to preclude having an explosive air. Under the condition of the influent nitrate surface loading of 1.04 g N/m(2)d, over 90% removal efficiencies of the total nitrogen and nitrate were achieved at the hydrogen pressure above 0.04 MPa. The results of denaturing gel gradient electrophoresis (DGGE), 16S rDNA gene sequence analysis, and hierarchical cluster analysis showed that the microbial community structures in MBfR were of low diversity, simple and stable at mature stages; and the beta-Proteobacteria, including Rhodocyclus, Hydrogenophaga, and beta-Proteobacteria HTCC379, probably play an important role in autohydrogenotrophic denitrification.

  9. Catalytic biofilms on structured packing for the production of glycolic acid.

    PubMed

    Li, Xuan Zhong; Hauer, Bernhard; Rosche, Bettina

    2013-02-01

    While structured packing modules are known to be efficient for surface wetting and gas-liquid exchange in abiotic surface catalysis, this model study explores structured packing as a growth surface for catalytic biofilms. Microbial biofilms have been proposed as self-immobilized and self-regenerating catalysts for the production of chemicals. A concern is that the complex and dynamic nature of biofilms may cause fluctuations in their catalytic performance over time or may affect process reproducibility. An aerated continuous trickle-bed biofilm reactor system was designed with a 3 L structured packing, liquid recycling and pH control. Pseudomonas diminuta established a biofilm on the stainless steel structured packing with a specific surface area of 500 m2 m-3 and catalyzed the oxidation of ethylene glycol to glycolic acid for over two months of continuous operation. A steady-state productivity of up to 1.6 gl-1h-1 was achieved at a dilution rate of 0.33 h-1. Process reproducibility between three independent runs was excellent, despite process interruptions and activity variations in cultures grown from biofilm effluent cells. The results demonstrate the robustness of a catalytic biofilm on structured packing, despite its dynamic nature. Implementation is recommended for whole-cell processes that require efficient gas-liquid exchange, catalyst retention for continuous operation, or improved catalyst stability.

  10. Measuring H(+) Pumping and Membrane Potential Formation in Sealed Membrane Vesicle Systems.

    PubMed

    Wielandt, Alex Green; Palmgren, Michael G; Fuglsang, Anja Thoe; Günther-Pomorski, Thomas; Justesen, Bo Højen

    2016-01-01

    The activity of enzymes involved in active transport of matter across lipid bilayers can conveniently be assayed by measuring their consumption of energy, such as ATP hydrolysis, while it is more challenging to directly measure their transport activities as the transported substrate is not converted into a product and only moves a few nanometers in space. Here, we describe two methods for the measurement of active proton pumping across lipid bilayers and the concomitant formation of a membrane potential, applying the dyes 9-amino-6-chloro-2-methoxyacridine (ACMA) and oxonol VI. The methods are exemplified by assaying transport of the Arabidopsis thaliana plasma membrane H(+)-ATPase (proton pump), which after heterologous expression in Saccharomyces cerevisiae and subsequent purification has been reconstituted in proteoliposomes.

  11. Synergistic effect of steam and lactic acid against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms on polyvinyl chloride and stainless steel.

    PubMed

    Ban, Ga-Hee; Park, Sang-Hyun; Kim, Sang-Oh; Ryu, Sangryeol; Kang, Dong-Hyun

    2012-07-02

    This study was designed to investigate the individual and combined effects of steam and lactic acid (LA) on the inactivation of biofilms formed by Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on polyvinyl chloride (PVC) and stainless steel. Six day old biofilms were developed on PVC and stainless steel coupons by using a mixture of three strains each of three foodborne pathogens at 25°C. After biofilm development, PVC and stainless steel coupons were treated with LA alone (immersed in 0.5% or 2% for 5s, 15s, and 30s), steam alone (on both sides for 5, 10, and 20s), and the combination of steam and LA. The numbers of biofilm cells of the three foodborne pathogens were significantly (p<0.05) reduced as the amount of LA and duration of steam exposure increased. There was a synergistic effect of steam and LA on the viability of biofilm cells of the three pathogens. For all biofilm cells of the three foodborne pathogens, reduction levels of individual treatments ranged from 0.11 to 2.12 log CFU/coupon. The combination treatment of steam and LA achieved an additional 0.2 to 2.11 log reduction compared to the sum of individual treatments. After a combined treatment of immersion in 2% LA for 15s or 30s followed by exposure to steam for 20s, biofilm cells of the three pathogens were reduced to below the detection limit (1.48 log). From the results of this study, bacterial populations of biofilms on PVC coupons did not receive the same thermal effect as on stainless steel coupons. Effectiveness of steam and LA may be attributed to the difference between Gram-negative and Gram-positive characteristics of the bacteria studied. The results of this study suggest that the combination of steam and LA has potential as a biofilm control intervention for food processing facilities. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. Immobilization of uranium in biofilm microorganisms exposed to groundwater seeps over granitic rock tunnel walls in Olkiluoto, Finland

    NASA Astrophysics Data System (ADS)

    Krawczyk-Bärsch, Evelyn; Lünsdorf, Heinrich; Pedersen, Karsten; Arnold, Thuro; Bok, Frank; Steudtner, Robin; Lehtinen, Anne; Brendler, Vinzenz

    2012-11-01

    In an underground rock characterization facility, the ONKALO tunnel in Finland, massive 5-10-mm thick biofilms were observed attached to tunnel walls where groundwater was seeping from bedrock fractures at a depth of 70 m. In laboratory experiments performed in a flow cell with detached biofilms to study the effect of uranium on the biofilm, uranium was added to the circulating groundwater (CGW) obtained from the fracture feeding the biofilm. The final uranium concentration in the CGW was adjusted to 4.25 × 10-5 M, in the range expected from a leaking spent nuclear fuel (SNF) canister in a future underground repository. The effects were investigated using microelectrodes to measure pH and Eh, time-resolved laser fluorescence spectroscopy (TRLFS), energy-filtered transmission electron microscopy (EF-TEM), and electron energy-loss spectroscopy (EELS) studies and thermodynamic calculations were utilized as well. The results indicated that the studied biofilms constituted their own microenvironments, which differed significantly from that of the CGW. A pH of 5.37 was recorded inside the biofilm, approximately 3.5 units lower than the pH observed in the CGW, due to sulfide oxidation to sulfuric acid in the biofilm. Similarly, the Eh of +73 mV inside the biofilm was approximately 420 mV lower than the Eh measured in the CGW. Adding uranium increased the pH in the biofilm to 7.27 and reduced the Eh to -164 mV. The changes of Eh and pH influenced the bioavailability of uranium, since microbial metabolic processes are sensitive to metals and their speciation. EF-TEM investigations indicated that uranium in the biofilm was immobilized intracellularly in microorganisms by the formation of metabolically mediated uranyl phosphate, similar to needle-shaped autunite (Ca[UO2]2[PO4]2·2-6H2O) or meta-autunite (Ca[UO2]2[PO4]2·10-12H2O). In contrast, TRLFS studies of the contaminated CGW identified aqueous uranium carbonate species, likely (Ca2UO2[CO3]3), formed due to the high

  13. Impact of ozonation, anion exchange resin and UV/H2O2 pre-treatments to control fouling of ultrafiltration membrane for drinking water treatment.

    PubMed

    Pramanik, Biplob Kumar; Pramanik, Sagor Kumar; Sarker, Dipok Chandra; Suja, Fatihah

    2017-06-01

    The effects of ozonation, anion exchange resin (AER) and UV/H 2 O 2 were investigated as a pre-treatment to control organic fouling (OF) of ultrafiltration membrane in the treatment of drinking water. It was found that high molecular weight (MW) organics such as protein and polysaccharide substances were majorly responsible for reversible fouling which contributed to 90% of total fouling. The decline rate increased with successive filtration cycles due to deposition of protein content over time. All pre-treatment could reduce the foulants of a Ultrafiltration membrane which contributed to the improvement in flux, and there was a greater improvement of flux by UV/H 2 O 2 (61%) than ozonation (43%) which in turn was greater than AER (23%) treatment. This was likely due to the effective removal/breakdown of high MW organic content. AER gave greater removal of biofouling potential components (such as biodegradable dissolved organic carbon and assimilable organic carbon contents) compared to UV/H 2 O 2 and ozonation treatment. Overall, this study demonstrated the potential of pre-treatments for reducing OF of ultrafiltration for the treatment of drinking water.

  14. Carbon dioxide (hydrogen sulfide) membrane separations and WGS membrane reactor modeling for fuel cells

    NASA Astrophysics Data System (ADS)

    Huang, Jin

    Acid-gas removal is of great importance in many environmental or energy-related processes. Compared to current commercial technologies, membrane-based CO2 and H2S capture has the advantages of low energy consumption, low weight and space requirement, simplicity of installation/operation, and high process flexibility. However, the large-scale application of the membrane separation technology is limited by the relatively low transport properties. In this study, CO2 (H2S)-selective polymeric membranes with high permeability and high selectivity have been studied based on the facilitated transport mechanism. The membrane showed facilitated effect for both CO2 and H2S. A CO2 permeability of above 2000 Barrers, a CO2/H2 selectivity of greater than 40, and a CO2/N2 selectivity of greater than 200 at 100--150°C were observed. As a result of higher reaction rate and smaller diffusing compound, the H2S permeability and H2S/H2 selectivity were about three times higher than those properties for CO2. The novel CO2-selective membrane has been applied to capture CO 2 from flue gas and natural gas. In the CO2 capture experiments from a gas mixture with N2 and H2, a permeate CO 2 dry concentration of greater than 98% was obtained by using steam as the sweep gas. In CO2/CH4 separation, decent CO 2 transport properties were obtained with a feed pressure up to 500 psia. With the thin-film composite membrane structure, significant increase on the CO2 flux was achieved with the decrease of the selective layer thickness. With the continuous removal of CO2, CO2-selective water-gas-shift (WGS) membrane reactor is a promising approach to enhance CO conversion and increase the purity of H2 at process pressure under relatively low temperature. The simultaneous reaction and transport process in the countercurrent WGS membrane reactor was simulated by using a one-dimensional non-isothermal model. The modeling results show that a CO concentration of less than 10 ppm and a H2 recovery of greater

  15. Linking ceragenins to water-treatment membranes to minimize biofouling.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hibbs, Michael R.; Altman, Susan Jeanne; Feng, Yanshu

    Ceragenins were used to create biofouling resistant water-treatment membranes. Ceragenins are synthetically produced antimicrobial peptide mimics that display broad-spectrum bactericidal activity. While ceragenins have been used on bio-medical devices, use of ceragenins on water-treatment membranes is novel. Biofouling impacts membrane separation processes for many industrial applications such as desalination, waste-water treatment, oil and gas extraction, and power generation. Biofouling results in a loss of permeate flux and increase in energy use. Creation of biofouling resistant membranes will assist in creation of clean water with lower energy usage and energy with lower water usage. Five methods of attaching three different cerageninmore » molecules were conducted and tested. Biofouling reduction was observed in the majority of the tests, indicating the ceragenins are a viable solution to biofouling on water treatment membranes. Silane direct attachment appears to be the most promising attachment method if a high concentration of CSA-121a is used. Additional refinement of the attachment methods are needed in order to achieve our goal of several log-reduction in biofilm cell density without impacting the membrane flux. Concurrently, biofilm forming bacteria were isolated from source waters relevant for water treatment: wastewater, agricultural drainage, river water, seawater, and brackish groundwater. These isolates can be used for future testing of methods to control biofouling. Once isolated, the ability of the isolates to grow biofilms was tested with high-throughput multiwell methods. Based on these tests, the following species were selected for further testing in tube reactors and CDC reactors: Pseudomonas ssp. (wastewater, agricultural drainage, and Colorado River water), Nocardia coeliaca or Rhodococcus spp. (wastewater), Pseudomonas fluorescens and Hydrogenophaga palleronii (agricultural drainage), Sulfitobacter donghicola, Rhodococcus fascians

  16. Photobleaching of red fluorescence in oral biofilms.

    PubMed

    Hope, C K; de Josselin de Jong, E; Field, M R T; Valappil, S P; Higham, S M

    2011-04-01

    Many species of oral bacteria can be induced to fluoresce due to the presence of endogenous porphyrins, a phenomenon that can be utilized to visualize and quantify dental plaque in the laboratory or clinical setting. However, an inevitable consequence of fluorescence is photobleaching, and the effects of this on longitudinal, quantitative analysis of dental plaque have yet to be ascertained. Filter membrane biofilms were grown from salivary inocula or single species (Prevotella nigrescens and Prevotella intermedia). The mature biofilms were then examined in a custom-made lighting rig comprising 405 nm light-emitting diodes capable of delivering 220 W/m(2) at the sample, an appropriate filter and a digital camera; a set-up analogous to quantitative light-induced fluorescence digital. Longitudinal sets of images were captured and processed to assess the degradation in red fluorescence over time. Photobleaching was observed in all instances. The highest rates of photobleaching were observed immediately after initiation of illumination, specifically during the first minute. Relative rates of photobleaching during the first minute of exposure were 19.17, 13.72 and 3.43 arbitrary units/min for P. nigrescens biofilms, microcosm biofilm and P. intermedia biofilms, respectively. Photobleaching could be problematic when making quantitative measurements of porphyrin fluorescence in situ. Reducing both light levels and exposure time, in combination with increased camera sensitivity, should be the default approach when undertaking analyses by quantitative light-induced fluorescence digital. © 2010 John Wiley & Sons A/S.

  17. Validation of the Zürich burn-biofilm model.

    PubMed

    Guggenheim, Merlin; Thurnheer, Thomas; Gmür, Rudolf; Giovanoli, Pietro; Guggenheim, Bernhard

    2011-11-01

    Despite advances in the use of topical and parenteral antimicrobial therapy and the practice of early tangential burn-wound excision, bacterial infection remains a major problem in the management of burn victims today. The purpose of this study was to design and evaluate a polyspecies biofilm model with bacteria known to cause severe infections in burn patients. The model is simple to prepare, maintain and analyse, and allows for short-term exposure to antimicrobials. Initial experiments showed that it was impossible to establish balanced polyspecies biofilms with an inoculum of Gram-positive and -negative bacteria. After 64.5 h of incubation, the Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) had suppressed the Gram-positives (Enterococcus faecalis, Staphylococcus aureus and Streptococcus intermedius). However, adding the Gram-negative bacteria after 41.5 h to an established biofilm of Gram-positives resulted in a balanced microbial consortium. After 64.5 h, all species were present in high numbers (10(7) to 10(8) colony forming units (CFU) per biofilm). Multiple repetitions showed high reproducibility of biofilm formation without significant differences between and within experiments. Combined fluorescence in situ hybridisation/confocal laser scanning microscopy (FISH/CLSM) analyses, for which biofilms had to be grown on a different non-flexible substrate (hydroxy apatite), revealed that, by 41.5 h, the biofilm consisted of an almost confluent layer of bacteria firmly adherent to the substratum. After 64.5 h (22 h after the addition of the Gram negatives), the biofilm consisted of a confluent mixture of single cells, an abundance of galaxies of bacteria with small lacunae and large amounts of extracellular matrix polysaccharides. The polyspecies biofilm model contains the most prevalent burn-associated Gram-positive and Gram-negative bacterial pathogens and mimics the Gram-negative shift observed in vivo. It shows excellent reproducibility

  18. Environmental and Genetic Factors Regulating Localization of the Plant Plasma Membrane H+-ATPase.

    PubMed

    Haruta, Miyoshi; Tan, Li Xuan; Bushey, Daniel B; Swanson, Sarah J; Sussman, Michael R

    2018-01-01

    A P-type H + -ATPase is the primary transporter that converts ATP to electrochemical energy at the plasma membrane of higher plants. Its product, the proton-motive force, is composed of an electrical potential and a pH gradient. Many studies have demonstrated that this proton-motive force not only drives the secondary transporters required for nutrient uptake, but also plays a direct role in regulating cell expansion. Here, we have generated a transgenic Arabidopsis ( Arabidopsis thaliana ) plant expressing H + -ATPase isoform 2 (AHA2) that is translationally fused with a fluorescent protein and examined its cellular localization by live-cell microscopy. Using a 3D imaging approach with seedlings grown for various times under a variety of light intensities, we demonstrate that AHA2 localization at the plasma membrane of root cells requires light. In dim light conditions, AHA2 is found in intracellular compartments, in addition to the plasma membrane. This localization profile was age-dependent and specific to cell types found in the transition zone located between the meristem and elongation zones. The accumulation of AHA2 in intracellular compartments is consistent with reduced H + secretion near the transition zone and the suppression of root growth. By examining AHA2 localization in a knockout mutant of a receptor protein kinase, FERONIA, we found that the intracellular accumulation of AHA2 in the transition zone is dependent on a functional FERONIA-dependent inhibitory response in root elongation. Overall, this study provides a molecular underpinning for understanding the genetic, environmental, and developmental factors influencing root growth via localization of the plasma membrane H + -ATPase. © 2018 American Society of Plant Biologists. All Rights Reserved.

  19. Plant lipid environment and membrane enzymes: the case of the plasma membrane H+-ATPase.

    PubMed

    Morales-Cedillo, Francisco; González-Solís, Ariadna; Gutiérrez-Angoa, Lizbeth; Cano-Ramírez, Dora Luz; Gavilanes-Ruiz, Marina

    2015-04-01

    Several lipid classes constitute the universal matrix of the biological membranes. With their amphipathic nature, lipids not only build the continuous barrier that confers identity to every cell and organelle, but they are also active actors that modulate the activity of the proteins immersed in the lipid bilayer. The plasma membrane H(+)-ATPase, an enzyme from plant cells, is an excellent example of a transmembrane protein whose activity is influenced by the hydrophilic compartments at both sides of the membrane and by the hydrophobic domains of the lipid bilayer. As a result, an extensive documentation of the effect of numerous amphiphiles in the enzyme activity can be found. Detergents, membrane glycerolipids, and sterols can produce activation or inhibition of the enzyme activity. In some cases, these effects are associated with the lipids of the membrane bulk, but in others, a direct interaction of the lipid with the protein is involved. This review gives an account of reports related to the action of the membrane lipids on the H(+)-ATPase activity.

  20. Na+/H+ Exchange Activity in the Plasma Membrane of Arabidopsis1

    PubMed Central

    Qiu, Quan-Sheng; Barkla, Bronwyn J.; Vera-Estrella, Rosario; Zhu, Jian-Kang; Schumaker, Karen S.

    2003-01-01

    In plants, Na+/H+ exchangers in the plasma membrane are critical for growth in high levels of salt, removing toxic Na+ from the cytoplasm by transport out of the cell. The molecular identity of a plasma membrane Na+/H+ exchanger in Arabidopsis (SOS1) has recently been determined. In this study, immunological analysis provided evidence that SOS1 localizes to the plasma membrane of leaves and roots. To characterize the transport activity of this protein, purified plasma membrane vesicles were isolated from leaves of Arabidopsis. Na+/H+ exchange activity, monitored as the ability of Na to dissipate an established pH gradient, was absent in plants grown without salt. However, exchange activity was induced when plants were grown in 250 mm NaCl and increased with prolonged salt exposure up to 8 d. H+-coupled exchange was specific for Na, because chloride salts of other monovalent cations did not dissipate the pH gradient. Na+/H+ exchange activity was dependent on Na (substrate) concentration, and kinetic analysis indicated that the affinity (apparent Km) of the transporter for Na+ is 22.8 mm. Data from two experimental approaches supports electroneutral exchange (one Na+ exchanged for one proton): (a) no change in membrane potential was measured during the exchange reaction, and (b) Na+/H+ exchange was unaffected by the presence or absence of a membrane potential. Results from this research provide a framework for future studies into the regulation of the plant plasma membrane Na+/H+ exchanger and its relative contribution to the maintenance of cellular Na+ homeostasis during plant growth in salt. PMID:12805632

  1. Silver colloidal nanoparticles: antifungal effect against adhered cells and biofilms of Candida albicans and Candida glabrata.

    PubMed

    Monteiro, D R; Gorup, L F; Silva, S; Negri, M; de Camargo, E R; Oliveira, R; Barbosa, D B; Henriques, M

    2011-08-01

    The aim of this study was to evaluate the effect of silver nanoparticles (SN) against Candida albicans and Candida glabrata adhered cells and biofilms. SN (average diameter 5 nm) were synthesized by silver nitrate reduction with sodium citrate and stabilized with ammonia. Minimal inhibitory concentration (MIC) tests were performed for C. albicans (n = 2) and C. glabrata (n = 2) grown in suspension following the Clinical Laboratory Standards Institute microbroth dilution method. SN were applied to adhered cells (2 h) or biofilms (48 h) and after 24 h of contact their effect was assessed by enumeration of colony forming units (CFUs) and quantification of total biomass (by crystal violet staining). The MIC results showed that SN were fungicidal against all strains tested at very low concentrations (0.4-3.3 μg ml(-1)). Furthermore, SN were more effective in reducing biofilm biomass when applied to adhered cells (2 h) than to pre-formed biofilms (48 h), with the exception of C. glabrata ATCC, which in both cases showed a reduction ∼90%. Regarding cell viability, SN were highly effective on adhered C. glabrata and respective biofilms. On C. albicans the effect was not so evident but there was also a reduction in the number of viable biofilm cells. In summary, SN may have the potential to be an effective alternative to conventional antifungal agents for future therapies in Candida-associated denture stomatitis.

  2. Efficacy of Ethanol against Candida albicans and Staphylococcus aureus Polymicrobial Biofilms

    PubMed Central

    Peters, Brian M.; Ward, Raven M.; Rane, Hallie S.; Lee, Samuel A.

    2013-01-01

    Candida albicans, an opportunistic fungus, and Staphylococcus aureus, a bacterial pathogen, are two clinically relevant biofilm-forming microbes responsible for a majority of catheter-related infections, with such infections often resulting in catheter loss and removal. Not only do these pathogens cause a substantial number of nosocomial infections independently, but also they are frequently found coexisting as polymicrobial biofilms on host and environmental surfaces. Antimicrobial lock therapy is a current strategy to sterilize infected catheters. However, the robustness of this technique against polymicrobial biofilms has remained largely untested. Due to its antimicrobial activity, safety, stability, and affordability, we tested the hypothesis that ethanol (EtOH) could serve as a potentially efficacious catheter lock solution against C. albicans and S. aureus biofilms. Therefore, we optimized the dose and time necessary to achieve killing of both monomicrobial and polymicrobial biofilms formed on polystyrene and silicone surfaces in a static microplate lock therapy model. Treatment with 30% EtOH for a minimum of 4 h was inhibitory for monomicrobial and polymicrobial biofilms, as evidenced by XTT {sodium 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide inner salt} metabolic activity assays and confocal microscopy. Experiments to determine the regrowth of microorganisms on silicone after EtOH treatment were also performed. Importantly, incubation with 30% EtOH for 4 h was sufficient to kill and inhibit the growth of C. albicans, while 50% EtOH was needed to completely inhibit the regrowth of S. aureus. In summary, we have systematically defined the dose and duration of EtOH treatment that are effective against and prevent regrowth of C. albicans and S. aureus monomicrobial and polymicrobial biofilms in an in vitro lock therapy model. PMID:23070170

  3. A Novel Antifungal Is Active against Candida albicans Biofilms and Inhibits Mutagenic Acetaldehyde Production In Vitro

    PubMed Central

    Nieminen, Mikko T.; Novak-Frazer, Lily; Rautemaa, Vilma; Rajendran, Ranjith; Sorsa, Timo; Ramage, Gordon; Bowyer, Paul; Rautemaa, Riina

    2014-01-01

    The ability of C. albicans to form biofilms is a major virulence factor and a challenge for management. This is evident in biofilm-associated chronic oral-oesophageal candidosis, which has been shown to be potentially carcinogenic in vivo. We have previously shown that most Candida spp. can produce significant levels of mutagenic acetaldehyde (ACH). ACH is also an important mediator of candidal biofilm formation. We have also reported that D,L-2-hydroxyisocaproic acid (HICA) significantly inhibits planktonic growth of C. albicans. The aim of the present study was to investigate the effect of HICA on C. albicans biofilm formation and ACH production in vitro. Inhibition of biofilm formation by HICA, analogous control compounds or caspofungin was measured using XTT to measure biofilm metabolic activity and PicoGreen as a marker of biomass. Biofilms were visualised by scanning electron microscopy (SEM). ACH levels were measured by gas chromatography. Transcriptional changes in the genes involved in ACH metabolism were measured using RT-qPCR. The mean metabolic activity and biomass of all pre-grown (4, 24, 48 h) biofilms were significantly reduced after exposure to HICA (p<0.05) with the largest reductions seen at acidic pH. Caspofungin was mainly active against biofilms pre-grown for 4 h at neutral pH. Mutagenic levels (>40 µM) of ACH were detected in 24 and 48 h biofilms at both pHs. Interestingly, no ACH production was detected from D-glucose in the presence of HICA at acidic pH (p<0.05). Expression of genes responsible for ACH catabolism was up-regulated by HICA but down-regulated by caspofungin. SEM showed aberrant hyphae and collapsed hyphal structures during incubation with HICA at acidic pH. We conclude that HICA has potential as an antifungal agent with ability to inhibit C. albicans cell growth and biofilm formation. HICA also significantly reduces the mutagenic potential of C. albicans biofilms, which may be important when treating bacterial-fungal biofilm

  4. The N-Terminus of Human Lactoferrin Displays Anti-biofilm Activity on Candida parapsilosis in Lumen Catheters.

    PubMed

    Fais, Roberta; Di Luca, Mariagrazia; Rizzato, Cosmeri; Morici, Paola; Bottai, Daria; Tavanti, Arianna; Lupetti, Antonella

    2017-01-01

    Candida parapsilosis is a major cause of hospital-acquired infection, often related to parenteral nutrition administered via catheters and hand colonization of health care workers, and its peculiar biofilm formation ability on plastic surfaces. The mortality rate of 30% points to the pressing need for new antifungal drugs. The present study aimed at analyzing the inhibitory activity of the N-terminal lactoferrin-derived peptide, further referred to as hLF 1-11, against biofilms produced by clinical isolates of C. parapsilosis characterized for their biofilm forming ability and fluconazole susceptibility. hLF 1-11 anti-biofilm activity was assessed in terms of reduction of biofilm biomass, metabolic activity, and observation of sessile cell morphology on polystyrene microtiter plates and using an in vitro model of catheter-associated C. parapsilosis biofilm production. Moreover, fluctuation in transcription levels of genes related to cell adhesion, hyphal development and extracellular matrix production upon peptide exposure were evaluated by quantitative real time RT-PCR. The results revealed that hLF 1-11 exhibits an inhibitory effect on biofilm formation by all the C. parapsilosis isolates tested, in a dose-dependent manner, regardless of their fluconazole susceptibility. In addition, hLF 1-11 induced a statistically significant dose-dependent reduction of preformed-biofilm cellular density and metabolic activity at high peptide concentrations only. Interestingly, when assessed in a catheter lumen, hLF 1-11 was able to induce a 2-log reduction of sessile cell viability at both the peptide concentrations used in RPMI diluted in NaPB. A more pronounced anti-biofilm effect was observed (3.5-log reduction) when a 10% glucose solution was used as experimental condition on both early and preformed C. parapsilosis biofilm. Quantitative real time RT-PCR experiments confirmed that hLF 1-11 down-regulates key biofilm related genes. The overall findings suggest hLF 1-11 as a

  5. The N-Terminus of Human Lactoferrin Displays Anti-biofilm Activity on Candida parapsilosis in Lumen Catheters

    PubMed Central

    Fais, Roberta; Di Luca, Mariagrazia; Rizzato, Cosmeri; Morici, Paola; Bottai, Daria; Tavanti, Arianna; Lupetti, Antonella

    2017-01-01

    Candida parapsilosis is a major cause of hospital-acquired infection, often related to parenteral nutrition administered via catheters and hand colonization of health care workers, and its peculiar biofilm formation ability on plastic surfaces. The mortality rate of 30% points to the pressing need for new antifungal drugs. The present study aimed at analyzing the inhibitory activity of the N-terminal lactoferrin-derived peptide, further referred to as hLF 1-11, against biofilms produced by clinical isolates of C. parapsilosis characterized for their biofilm forming ability and fluconazole susceptibility. hLF 1-11 anti-biofilm activity was assessed in terms of reduction of biofilm biomass, metabolic activity, and observation of sessile cell morphology on polystyrene microtiter plates and using an in vitro model of catheter-associated C. parapsilosis biofilm production. Moreover, fluctuation in transcription levels of genes related to cell adhesion, hyphal development and extracellular matrix production upon peptide exposure were evaluated by quantitative real time RT-PCR. The results revealed that hLF 1-11 exhibits an inhibitory effect on biofilm formation by all the C. parapsilosis isolates tested, in a dose-dependent manner, regardless of their fluconazole susceptibility. In addition, hLF 1-11 induced a statistically significant dose-dependent reduction of preformed-biofilm cellular density and metabolic activity at high peptide concentrations only. Interestingly, when assessed in a catheter lumen, hLF 1-11 was able to induce a 2-log reduction of sessile cell viability at both the peptide concentrations used in RPMI diluted in NaPB. A more pronounced anti-biofilm effect was observed (3.5-log reduction) when a 10% glucose solution was used as experimental condition on both early and preformed C. parapsilosis biofilm. Quantitative real time RT-PCR experiments confirmed that hLF 1-11 down-regulates key biofilm related genes. The overall findings suggest hLF 1-11 as a

  6. Strain-level genetic diversity of Methylophaga nitratireducenticrescens confers plasticity to denitrification capacity in a methylotrophic marine denitrifying biofilm.

    PubMed

    Geoffroy, Valérie; Payette, Geneviève; Mauffrey, Florian; Lestin, Livie; Constant, Philippe; Villemur, Richard

    2018-01-01

    The biofilm of a methanol-fed, fluidized denitrification system treating a marine effluent is composed of multi-species microorganisms, among which Hyphomicrobium nitrativorans NL23 and Methylophaga nitratireducenticrescens JAM1 are the principal bacteria involved in the denitrifying activities. Strain NL23 can carry complete nitrate (NO[Formula: see text]) reduction to N 2 , whereas strain JAM1 can perform 3 out of the 4 reduction steps. A small proportion of other denitrifiers exists in the biofilm, suggesting the potential plasticity of the biofilm in adapting to environmental changes. Here, we report the acclimation of the denitrifying biofilm from continuous operating mode to batch operating mode, and the isolation and characterization from the acclimated biofilm of a new denitrifying bacterial strain, named GP59. The denitrifying biofilm was batch-cultured under anoxic conditions. The acclimated biofilm was plated on Methylophaga specific medium to isolate denitrifying Methylophaga isolates. Planktonic cultures of strains GP59 and JAM1 were performed, and the growth and the dynamics of NO[Formula: see text], nitrite (NO[Formula: see text]) and N 2 O were determined. The genomes of strains GP59 and JAM1 were sequenced and compared. The transcriptomes of strains GP59 and JAM1 were derived from anoxic cultures. During batch cultures of the biofilm, we observed the disappearance of H. nitrativorans NL23 without affecting the denitrification performance. From the acclimated biofilm, we isolated strain GP59 that can perform, like H. nitrativorans NL23, the complete denitrification pathway. The GP59 cell concentration in the acclimated biofilm was 2-3 orders of magnitude higher than M. nitratireducenticrescens JAM1 and H. nitrativorans NL23. Genome analyses revealed that strain GP59 belongs to the species M. nitratireducenticrescens . The GP59 genome shares more than 85% of its coding sequences with those of strain JAM1. Based on transcriptomic analyses of anoxic

  7. Fatty acid fouling of forward osmosis membrane: Effects of pH, calcium, membrane orientation, initial permeate flux and foulant composition.

    PubMed

    Zhao, Pin; Gao, Baoyu; Yue, Qinyan; Liu, Pan; Shon, Ho Kyong

    2016-08-01

    Octanoic acid (OA) was selected to represent fatty acids in effluent organic matter (EOM). The effects of feed solution (FS) properties, membrane orientation and initial permeate flux on OA fouling in forward osmosis (FO) were investigated. The undissociated OA formed a cake layer quickly and caused the water flux to decline significantly in the initial 0.5hr at unadjusted pH3.56; while the fully dissociated OA behaved as an anionic surfactant and promoted the water permeation at an elevated pH of 9.00. Moreover, except at the initial stage, the sudden decline of water flux (meaning the occurrence of severe membrane fouling) occurred in two conditions: 1. 0.5mmol/L Ca(2+), active layer facing draw solution (AL-DS) and 1.5mol/L NaCl (DS); 2. No Ca(2+), active layer-facing FS (AL-FS) and 4mol/L NaCl (DS). This demonstrated that cake layer compaction or pore blocking occurred only when enough foulants were absorbed into the membrane surface, and the water permeation was high enough to compact the deposit inside the porous substrate. Furthermore, bovine serum albumin (BSA) was selected as a co-foulant. The water flux of both co-foulants was between the fluxes obtained separately for the two foulants at pH3.56, and larger than the two values at pH9.00. This manifested that, at pH3.56, BSA alleviated the effect of the cake layer caused by OA, and OA enhanced BSA fouling simultaneously; while at pH9.00, the mutual effects of OA and BSA eased the membrane fouling. Copyright © 2016. Published by Elsevier B.V.

  8. Application of bacteriophages to reduce biofilms formed by hydrogen sulfide producing bacteria on surfaces in a rendering plant.

    PubMed

    Gong, Chao; Jiang, Xiuping

    2015-08-01

    Hydrogen sulfide producing bacteria (SPB) in raw animal by-products are likely to grow and form biofilms in the rendering processing environments, resulting in the release of harmful hydrogen sulfide (H2S) gas. The objective of this study was to reduce SPB biofilms formed on different surfaces typically found in rendering plants by applying a bacteriophage cocktail. Using a 96-well microplate method, we determined that 3 SPB strains of Citrobacter freundii and Hafnia alvei are strong biofilm formers. Application of 9 bacteriophages (10(7) PFU/mL) from families of Siphoviridae and Myoviridae resulted in a 33%-70% reduction of biofilm formation by each SPB strain. On stainless steel and plastic templates, phage treatment (10(8) PFU/mL) reduced the attached cells of a mixed SPB culture (no biofilm) by 2.3 and 2.7 log CFU/cm(2) within 6 h at 30 °C, respectively, as compared with 2 and 1.5 log CFU/cm(2) reductions of SPB biofilms within 6 h at 30 °C. Phage treatment was also applied to indigenous SPB biofilms formed on the environmental surface, stainless steel, high-density polyethylene plastic, and rubber templates in a rendering plant. With phage treatment (10(9) PFU/mL), SPB biofilms were reduced by 0.7-1.4, 0.3-0.6, and 0.2-0.6 log CFU/cm(2) in spring, summer, and fall trials, respectively. Our study demonstrated that bacteriophages could effectively reduce the selected SPB strains either attached to or in formed biofilms on various surfaces and could to some extent reduce the indigenous SPB biofilms on the surfaces in the rendering environment.

  9. Desferrioxamine as an electron donor. Inhibition of membranal lipid peroxidation initiated by H2O2-activated metmyoglobin and other peroxidizing systems.

    PubMed

    Kanner, J; Harel, S

    1987-01-01

    Desferrioxamine (DFO) involvement in several peroxidative systems was studied. These systems included: a) membranal lipid peroxidation initiated by H2O2-activated metmyoglobin (or methemoglobin); b) phenol-red oxidation by activated metmyoglobin or horseradish peroxidase (HRP): c) beta-carotene-linoleate couple oxidation stimulated by lipoxygenase or hemin. Desferrioxamine was found to inhibit all these systems but not ferrioxamine (FO). Phenol-red oxidation by H2O2-horseradish peroxidase was inhibited competitively with DFO. Kinetic studies using the spectra changes in the Soret region of metmyoglobin suggest a mechanism by which H2O2 reacts with the iron-heme to form an intermediate of oxy-ferryl myoglobin that subsequently reacts with DFO to return the activated compound to the resting state. These activities of DFO resemble the reaction of other electron donors.

  10. Phylogenetic Evidence for H2 based Electron Bifurcation In Early Life

    NASA Astrophysics Data System (ADS)

    Adams, M. W.; Boyd, E. S.; Schut, G.; Peters, J.

    2012-12-01

    Energy conservation is a fundamental underpinning of all life and is accomplished by electron transport phosphorylation and/or substrate level phosphorylation. A third mechanism, known as flavin-based electron bifurcation, has recently been established as a mechanism by which life can conserve energy. In this mechanism, a flavin-containing multienzyme complex catalyzes the thermodynamically unfavorable reduction of low potential ferredoxin using electron donors with higher potentials, such as NAD(P)H or H2. Such endergonic reactions are driven forward through the simultaneous oxidation of the electron donor with higher potential acceptors such as NAD+ or heterodisulfide. Membrane associated energy converting [NiFe]hydrogenases (Ech, Eha) link the oxidation of ferredoxin with the production of H2 and in the process conserve energy in the form of an ion (Na+/H+) gradient. Ech/Eha exhibit a modular composition represented by a Na+/H+ antiporter domain and a [NiFe] hydrogenase domain. In addition, Ech/Eha can be accompanied by a formate dehyrogenase, carbon monoxide dehydrogenase, or an FAD/NAD(P)H module that enables coupling with these substrates. Representatives of Ech/Eha have been identified among anaerobic Archaea and Bacteria, including deeply rooted methanogens, sulfur-reducing Crenarcheota/Euryarchaeota as well as Thermotogae. Ech exhibit extensive homology to a number of subunits within the NADH quinone oxidoreductase or complex I family (Nuo, Fpo). Metabolically, Ech generally couple the oxidation of carbon monooxide, formate or ferredoxin to the production of H2. In contrast, the Eha complex couples the translocation of Na+ and the oxidation of H2 to the reduction of ferredoxin, which is then available for the reduction of CO2 in methanogens. In the case of Eha, the gradient of Na+/H+ produced through translocation coupled to ferredoxin oxidation can in be used to drive the phosphorylation of ADP via an ATP synthase complex, thereby representing one of the

  11. The Xylella fastidiosa PD1063 Protein Is Secreted in Association with Outer Membrane Vesicles

    PubMed Central

    Pierce, Brittany K.; Voegel, Tanja; Kirkpatrick, Bruce C.

    2014-01-01

    Xylella fastidiosa is a gram-negative, xylem-limited plant pathogenic bacterium that causes disease in a variety of economically important agricultural crops including Pierce's disease of grapevines. Xylella fastidiosa biofilms formed in the xylem vessels of plants play a key role in early colonization and pathogenicity by providing a protected niche and enhanced cell survival. Here we investigate the role of Xylella fastidiosa PD1063, the predicted ortholog of Xanthomonas oryzae pv. oryzae PXO_03968, which encodes an outer membrane protein. To assess the function of the Xylella fastidiosa ortholog, we created Xylella fastidiosa mutants deleted for PD1063 and then assessed biofilm formation, cell-cell aggregation and cell growth in vitro. We also assessed disease severity and pathogen titers in grapevines mechanically inoculated with the Xylella fastidiosa PD1063 mutant. We found a significant decrease in cell-cell aggregation among PD1063 mutants but no differences in cell growth, biofilm formation, disease severity or titers in planta. Based on the demonstration that Xanthomonas oryzae pv. oryzae PXO_03968 encodes an outer membrane protein, secreted in association with outer membrane vesicles, we predicted that PD1063 would also be secreted in a similar manner. Using anti-PD1063 antibodies, we found PD1063 in the supernatant and secreted in association with outer membrane vesicles. PD1063 purified from the supernatant, outer membrane fractions and outer membrane vesicles was 19.2 kD, corresponding to the predicted size of the processed protein. Our findings suggest Xylella fastidiosa PD1063 is not essential for development of Pierce's disease in Vitis vinifera grapevines although further research is required to determine the function of the PD1063 outer membrane protein in Xylella fastidiosa. PMID:25426629

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

    PubMed

    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.

  13. Modelling bio-electrosynthesis in a reverse microbial fuel cell to produce acetate from CO2 and H2O.

    PubMed

    Kazemi, M; Biria, D; Rismani-Yazdi, H

    2015-05-21

    Bio-electrosynthesis is one of the significant developments in reverse microbial fuel cell technology which is potentially capable of creating organic compounds by combining CO2 with H2O. Accordingly, the main objective in the current study was to present a model of microbial electrosynthesis for producing organic compounds (acetate) based on direct conduction of electrons in biofilms. The proposed model enjoys a high degree of rigor because it can predict variations in the substrate concentration, electrical potential, current density and the thickness of the biofilm. Additionally, coulombic efficiency was investigated as a function of substrate concentration and cathode potential. For a system containing CO2 as the substrate and Sporomusa ovata as the biofilm forming microorganism, an increase in the substrate concentration at a constant potential can lead to a decrease in coulombic efficiency as well as an increase in current density and biofilm thickness. On the other hand, an increase in the surface cathodic voltage at a constant substrate concentration may result in an increase in the coulombic efficiency and a decrease in the current density. The maximum coulombic efficiency was revealed to be 75% at a substrate concentration of 0.025 mmol cm(-3) and 55% at a surface cathodic voltage of -0.3 V producing a high range of acetate production by creating an optimal state in the concentration and potential intervals. Finally, the validity of the model was verified by comparing the obtained results with related experimental findings.

  14. The CsgA and Lpp proteins affect HEp-2 cell invasion, motility, and biofilm formation in a strain of Escherichia coli O157:H7

    USDA-ARS?s Scientific Manuscript database

    In Escherichia coli O157:H7 strain ATCC 43895, a guanine to thymine transversion in the csgD promoter created strain 43895OR. Strain 43895OR produces an abundant extracellular matrix rich in curli fibers, forms biofilm on solid surfaces, invades cultured epithelial cells, and is more virulent in mic...

  15. Comprehensive Study on Ceramic Membranes for Low-Cost Microbial Fuel Cells.

    PubMed

    Pasternak, Grzegorz; Greenman, John; Ieropoulos, Ioannis

    2016-01-08

    Microbial fuel cells (MFCs) made with different types of ceramic membranes were investigated to find a low-cost alternative to commercially available proton exchange membranes. The MFCs operated with fresh human urine as the fuel. Pyrophyllite and earthenware produced the best performance to reach power densities of 6.93 and 6.85 W m(-3), respectively, whereas mullite and alumina achieved power densities of 4.98 and 2.60 W m(-3), respectively. The results indicate the dependence of bio-film growth and activity on the type of ceramic membrane applied. The most favourable conditions were created in earthenware MFCs. The performance of the ceramic membranes was related to their physical and chemical properties determined by environmental scanning electron microscopy and energy dispersive X-ray spectroscopy. The cost of mullite, earthenware, pyrophyllite and alumina was estimated to be 13.61, 4.14, 387.96 and 177.03 GBP m(-2), respectively. The results indicate that earthenware and mullite are good substitutes for commercially available proton exchange membranes, which makes the MFC technology accessible in developing countries. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Nanotechnology-based drug delivery systems for control of microbial biofilms: a review.

    PubMed

    Dos Santos Ramos, Matheus Aparecido; Da Silva, Patrícia Bento; Spósito, Larissa; De Toledo, Luciani Gaspar; Bonifácio, Bruna Vidal; Rodero, Camila Fernanda; Dos Santos, Karen Cristina; Chorilli, Marlus; Bauab, Taís Maria

    2018-01-01

    Since the dawn of civilization, it has been understood that pathogenic microorganisms cause infectious conditions in humans, which at times, may prove fatal. Among the different virulent properties of microorganisms is their ability to form biofilms, which has been directly related to the development of chronic infections with increased disease severity. A problem in the elimination of such complex structures (biofilms) is resistance to the drugs that are currently used in clinical practice, and therefore, it becomes imperative to search for new compounds that have anti-biofilm activity. In this context, nanotechnology provides secure platforms for targeted delivery of drugs to treat numerous microbial infections that are caused by biofilms. Among the many applications of such nanotechnology-based drug delivery systems is their ability to enhance the bioactive potential of therapeutic agents. The present study reports the use of important nanoparticles, such as liposomes, microemulsions, cyclodextrins, solid lipid nanoparticles, polymeric nanoparticles, and metallic nanoparticles, in controlling microbial biofilms by targeted drug delivery. Such utilization of these nanosystems has led to a better understanding of their applications and their role in combating biofilms.

  17. The biofilm matrix destabilizers, EDTA and DNaseI, enhance the susceptibility of nontypeable Hemophilus influenzae biofilms to treatment with ampicillin and ciprofloxacin.

    PubMed

    Cavaliere, Rosalia; Ball, Jessica L; Turnbull, Lynne; Whitchurch, Cynthia B

    2014-08-01

    Nontypeable Hemophilus influenzae (NTHi) is a Gram-negative bacterial pathogen that causes chronic biofilm infections of the ears and airways. The biofilm matrix provides structural integrity to the biofilm and protects biofilm cells from antibiotic exposure by reducing penetration of antimicrobial compounds into the biofilm. Extracellular DNA (eDNA) has been found to be a major matrix component of biofilms formed by many species of Gram-positive and Gram-negative bacteria, including NTHi. Interestingly, the cation chelator ethylenediaminetetra-acetic acid (EDTA) has been shown to reduce the matrix strength of biofilms of several bacterial species as well as to have bactericidal activity against various pathogens. EDTA exerts its antimicrobial activity by chelating divalent cations necessary for growth and membrane stability and by destabilizing the matrix thus enhancing the detachment of bacterial cells from the biofilm. In this study, we have explored the role of divalent cations in NTHi biofilm development and stability. We have utilized in vitro static and continuous flow models of biofilm development by NTHi to demonstrate that magnesium cations enhance biofilm formation by NTHi. We found that the divalent cation chelator EDTA is effective at both preventing NTHi biofilm formation and at treating established NTHi biofilms. Furthermore, we found that the matrix destablilizers EDTA and DNaseI increase the susceptibility of NTHi biofilms to ampicillin and ciprofloxacin. Our observations indicate that DNaseI and EDTA enhance the efficacy of antibiotic treatment of NTHi biofilms. These observations may lead to new strategies that will improve the treatment options available to patients with chronic NTHi infections. © 2014 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  18. Potential Antifungal Targets against a Candida Biofilm Based on an Enzyme in the Arachidonic Acid Cascade—A Review

    PubMed Central

    Liu, Xinning; Wang, Decai; Yu, Cuixiang; Li, Tao; Liu, Jianqiao; Sun, Shujuan

    2016-01-01

    Candida is an important opportunistic fungal pathogen, especially in biofilm associated infections. The formation of a Candida biofilm can decrease Candida sensitivity to antifungal drugs and cause drug resistance. Although many effective antifungal drugs are available, their applications are limited due to their high toxicity and cost. Seeking new antifungal agents that are effective against biofilm-associated infection is an urgent need. Many research efforts are underway, and some progress has been made in this field. It has been shown that the arachidonic acid cascade plays an important role in fungal morphogenesis and pathogenicity. Notably, prostaglandin E2 (PGE2) can promote the formation of a Candida biofilm. Recently, the inhibition of PGE2 has received much attention. Studies have shown that cyclooxygenase (COX) inhibitors, such as aspirin, ibuprofen, and indomethacin, combined with fluconazole can significantly reduce Candida adhesion and biofilm development and increase fluconazole susceptibility; the MIC of fluconazole can be decrease from 64 to 2 μg/ml when used in combination with ibuprofen. In addition, in vivo studies have also confirmed the antifungal activities of these inhibitors. In this article, we mainly review the relationship between PGE2 and Candida biofilm, summarize the antifungal activities of COX inhibitors and analyze the possible antifungal activity of microsomal prostaglandin E synthase-1 (MPGES-1) inhibitors; additionally, other factors that influence PGE2 production are also discussed. Hopefully this review can disclose potential antifungal targets based on the arachidonic acid cascade and provide a prevailing strategy to alleviate Candida albicans biofilm formation. PMID:27999568

  19. Disinfection of Streptococcus mutans biofilm by a non-thermal atmospheric plasma brush

    NASA Astrophysics Data System (ADS)

    Hong, Qing; Dong, Xiaoqing; Chen, Meng; Xu, Yuanxi; Sun, Hongmin; Hong, Liang; Wang, Yong; Yu, Qingsong

    2016-07-01

    This study investigated the argon plasma treatment effect on disinfecting dental biofilm by using an atmospheric pressure plasma brush. Streptococcus mutans biofilms were developed for 3 days on the surfaces of hydroxyapatite (HA) discs, which were used to simulate human tooth enamel. After plasma treatment, cell viability in the S. mutans biofilms was characterized by using 3-(4,5-dimethylazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and confocal laser scanning microscopy (CLSM). Compared with the untreated control group, about 90% bacterial reduction in the biofilms was observed after 1 min plasma treatment. Scanning electron microscopy (SEM) examination indicated severe cell damages occurred on the top surface of the plasma treated biofilms. Confocal laser scanning microscopy (CLSM) showed that plasma treatment was effective as deep as 20 µm into the biofilms. When combined with antibiotic treatment using 0.2% chlorhexidine digluconate solution, the plasma treatment became more effective and over 96% bacterial reduction was observed with 1 min plasma treatment.

  20. Morphomechanics of bacterial biofilms undergoing anisotropic differential growth

    NASA Astrophysics Data System (ADS)

    Zhang, Cheng; Li, Bo; Huang, Xiao; Ni, Yong; Feng, Xi-Qiao

    2016-10-01

    Growing bacterial biofilms exhibit a number of surface morphologies, e.g., concentric wrinkles, radial ridges, and labyrinthine networks, depending on their physiological status and nutrient access. We explore the mechanisms underlying the emergence of these greatly different morphologies. Ginzburg-Landau kinetic method and Fourier spectral method are integrated to simulate the morphological evolution of bacterial biofilms. It is shown that the morphological instability of biofilms is triggered by the stresses induced by anisotropic and heterogeneous bacterial expansion, and involves the competition between membrane energy and bending energy. Local interfacial delamination further enriches the morphologies of biofilms. Phase diagrams are established to reveal how the anisotropy and spatial heterogeneity of growth modulate the surface patterns. The mechanics of three-dimensional microbial morphogenesis may also underpin self-organization in other development systems and provide a potential strategy for engineering microscopic structures from bacterial aggregates.