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Sample records for geobacter metallireducens gs15

  1. The genome sequence of Geobacter metallireducens: features of metabolism, physiology and regulation common and dissimilar to Geobacter sulfurreducens

    Energy Technology Data Exchange (ETDEWEB)

    Aklujkar, Muktak; Krushkal, Julia; DiBartolo, Genevieve; Lapidus, Alla; Land, Miriam L.; Lovley, Derek R.

    2008-12-01

    Background: The genome sequence of Geobacter metallireducens is the second to be completed from the metal-respiring genus Geobacter, and is compared in this report to that of Geobacter sulfurreducens in order to understand their metabolic, physiological and regulatory similarities and differences. Results: The experimentally observed greater metabolic versatility of G. metallireducens versus G. sulfurreducens is borne out by the presence of more numerous genes for metabolism of organic acids including acetate, propionate, and pyruvate. Although G. metallireducens lacks a dicarboxylic acid transporter, it has acquired a second succinate dehydrogenase/fumarate reductase complex, suggesting that respiration of fumarate was important until recently in its evolutionary history. Vestiges of the molybdate (ModE) regulon of G. sulfurreducens can be detected in G. metallireducens, which has lost the global regulatory protein ModE but retained some putative ModE-binding sites and multiplied certain genes of molybdenum cofactor biosynthesis. Several enzymes of amino acid metabolism are of different origin in the two species, but significant patterns of gene organization are conserved. Whereas most Geobacteraceae are predicted to obtain biosynthetic reducing equivalents from electron transfer pathways via a ferredoxin oxidoreductase, G. metallireducens can derive them from the oxidative pentose phosphate pathway. In addition to the evidence of greater metabolic versatility, the G. metallireducens genome is also remarkable for the abundance of multicopy nucleotide sequences found in intergenic regions and even within genes. Conclusion: The genomic evidence suggests that metabolism, physiology and regulation of gene expression in G. metallireducens may be dramatically different from other Geobacteraceae.

  2. Flux analysis of central metabolic pathways in the Fe(III)-reducing organism Geobacter metallireducens via 13C isotopiclabeling

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Yinjie J.; Chakraborty, Romy; Martin, Hector Garcia; Chu,Jeannie; Hazen, Terry C.; Keasling, Jay D.

    2007-08-13

    We analyzed the carbon fluxes in the central metabolism ofGeobacter metallireducens strain GS-15 using 13C isotopomer modeling.Acetate labeled in the 1st or 2nd position was the sole carbon source,and Fe-NTA was the sole terminal electron acceptor. The measured labeledacetate uptake rate was 21 mmol/gdw/h in the exponential growth phase.The resulting isotope labeling pattern of amino acids allowed an accuratedetermination of the in vivo global metabolic reaction rates (fluxes)through the central metabolic pathways using a computational isotopomermodel. The model indicated that over 90 percent of the acetate wascompletely oxidized to CO2 via a complete tricarboxylic acid (TCA) cyclewhile reducing iron. Pyruvate carboxylase and phosphoenolpyruvatecarboxykinase were present under these conditions, but enzymes in theglyoxylate shunt and malic enzyme were absent. Gluconeogenesis and thepentose phosphate pathway were mainly employed for biosynthesis andaccounted for less than 3 percent of total carbon consumption. The modelalso indicated surprisingly high reversibility in the reaction betweenoxoglutarate and succinate. This step operates close to the thermodynamicequilibrium possibly because succinate is synthesized via a transferasereaction, and its product, acetyl-CoA, inhibits the conversion ofoxoglutarate to succinate. These findings enable a better understandingof the relationship between genome annotation and extant metabolicpathways in G. metallireducens.

  3. Identification of genes specifically required for the anaerobic metabolism of benzene in Geobacter metallireducens

    DEFF Research Database (Denmark)

    Zhang, Tian; Tremblay, Pier-Luc; Chaurasia, Akhilesh Kumar

    2014-01-01

    Although the biochemical pathways for the anaerobic degradation of many of the hydrocarbon constituents in petroleum reservoirs have been elucidated, the mechanisms for anaerobic activation of benzene, a very stable molecule, are not known. Previous studies have demonstrated that Geobacter...... metallireducens can anaerobically oxidize benzene to carbon dioxide with Fe(III) as the sole electron acceptor and that phenol is an intermediate in benzene oxidation. In an attempt to identify enzymes that might be involved in the conversion of benzene to phenol, whole-genome gene transcript abundance...... was compared in cells metabolizing benzene and cells metabolizing phenol. Eleven genes had significantly higher transcript abundance in benzene-metabolizing cells. Five of these genes had annotations suggesting that they did not encode proteins that could be involved in benzene metabolism and were not further...

  4. Genomic and microarray analysis of aromatics degradation in Geobacter metallireducens and comparison to a Geobacter isolate from a contaminated field site

    Directory of Open Access Journals (Sweden)

    Zhou Jizhong

    2007-06-01

    Full Text Available Abstract Background Groundwater and subsurface environments contaminated with aromatic compounds can be remediated in situ by Geobacter species that couple oxidation of these compounds to reduction of Fe(III-oxides. Geobacter metallireducens metabolizes many aromatic compounds, but the enzymes involved are not well known. Results The complete G. metallireducens genome contained a 300 kb island predicted to encode enzymes for the degradation of phenol, p-cresol, 4-hydroxybenzaldehyde, 4-hydroxybenzoate, benzyl alcohol, benzaldehyde, and benzoate. Toluene degradation genes were encoded in a separate region. None of these genes was found in closely related species that cannot degrade aromatic compounds. Abundant transposons and phage-like genes in the island suggest mobility, but nucleotide composition and lack of synteny with other species do not suggest a recent transfer. The inferred degradation pathways are similar to those in species that anaerobically oxidize aromatic compounds with nitrate as an electron acceptor. In these pathways the aromatic compounds are converted to benzoyl-CoA and then to 3-hydroxypimelyl-CoA. However, in G. metallireducens there were no genes for the energetically-expensive dearomatizing enzyme. Whole-genome changes in transcript levels were identified in cells oxidizing benzoate. These supported the predicted pathway, identified induced fatty-acid oxidation genes, and identified an apparent shift in the TCA cycle to a putative ATP-yielding succinyl-CoA synthase. Paralogs to several genes in the pathway were also induced, as were several putative molybdo-proteins. Comparison of the aromatics degradation pathway genes to the genome of an isolate from a contaminated field site showed very similar content, and suggested this strain degrades many of the same compounds. This strain also lacked a classical dearomatizing enzyme, but contained two copies of an eight-gene cluster encoding redox proteins that was 30-fold

  5. Adaptation of the Biolog Phenotype MicroArrayTM Technology to Profile the Obligate Anaerobe Geobacter metallireducens

    Energy Technology Data Exchange (ETDEWEB)

    Joyner, Dominique; Fortney, Julian; Chakraborty, Romy; Hazen, Terry

    2010-05-17

    The Biolog OmniLog? Phenotype MicroArray (PM) plate technology was successfully adapted to generate a select phenotypic profile of the strict anaerobe Geobacter metallireducens (G.m.). The profile generated for G.m. provides insight into the chemical sensitivity of the organism as well as some of its metabolic capabilities when grown with a basal medium containing acetate and Fe(III). The PM technology was developed for aerobic organisms. The reduction of a tetrazolium dye by the test organism represents metabolic activity on the array which is detected and measured by the OmniLog(R) system. We have previously adapted the technology for the anaerobic sulfate reducing bacterium Desulfovibrio vulgaris. In this work, we have taken the technology a step further by adapting it for the iron reducing obligate anaerobe Geobacter metallireducens. In an osmotic stress microarray it was determined that the organism has higher sensitivity to impermeable solutes 3-6percent KCl and 2-5percent NaNO3 that result in osmotic stress by osmosis to the cell than to permeable non-ionic solutes represented by 5-20percent ethylene glycol and 2-3percent urea. The osmotic stress microarray also includes an array of osmoprotectants and precursor molecules that were screened to identify substrates that would provide osmotic protection to NaCl stress. None of the substrates tested conferred resistance to elevated concentrations of salt. Verification studies in which G.m. was grown in defined medium amended with 100mM NaCl (MIC) and the common osmoprotectants betaine, glycine and proline supported the PM findings. Further verification was done by analysis of transcriptomic profiles of G.m. grown under 100mM NaCl stress that revealed up-regulation of genes related to degradation rather than accumulation of the above-mentioned osmoprotectants. The phenotypic profile, supported by additional analysis indicates that the accumulation of these osmoprotectants as a response to salt stress does not

  6. Direct interspecies electron transfer between Geobacter metallireducens and Methanosarcina barkeri

    DEFF Research Database (Denmark)

    Rotaru, Amelia-Elena; Shrestha, Pravin Malla; Liu, Fanghua

    2014-01-01

    of granular activated carbon permitted the pilin-deficient G. metallireducens to share electrons with M. barkeri, demonstrating that this conductive material could substitute for pili in promoting DIET. When M. barkeri was grown in co-culture with the H2-producing Pelobacter carbinolicus, incapable of DIET, M...

  7. Evaluation of a Genome-Scale In Silico Metabolic Model for Geobacter metallireducens by Using Proteomic Data from a Field Biostimulation Experiment

    Science.gov (United States)

    Fang, Yilin; Yabusaki, Steven B.; Lipton, Mary S.; Long, Philip E.

    2012-01-01

    Accurately predicting the interactions between microbial metabolism and the physical subsurface environment is necessary to enhance subsurface energy development, soil and groundwater cleanup, and carbon management. This study was an initial attempt to confirm the metabolic functional roles within an in silico model using environmental proteomic data collected during field experiments. Shotgun global proteomics data collected during a subsurface biostimulation experiment were used to validate a genome-scale metabolic model of Geobacter metallireducens—specifically, the ability of the metabolic model to predict metal reduction, biomass yield, and growth rate under dynamic field conditions. The constraint-based in silico model of G. metallireducens relates an annotated genome sequence to the physiological functions with 697 reactions controlled by 747 enzyme-coding genes. Proteomic analysis showed that 180 of the 637 G. metallireducens proteins detected during the 2008 experiment were associated with specific metabolic reactions in the in silico model. When the field-calibrated Fe(III) terminal electron acceptor process reaction in a reactive transport model for the field experiments was replaced with the genome-scale model, the model predicted that the largest metabolic fluxes through the in silico model reactions generally correspond to the highest abundances of proteins that catalyze those reactions. Central metabolism predicted by the model agrees well with protein abundance profiles inferred from proteomic analysis. Model discrepancies with the proteomic data, such as the relatively low abundances of proteins associated with amino acid transport and metabolism, revealed pathways or flux constraints in the in silico model that could be updated to more accurately predict metabolic processes that occur in the subsurface environment. PMID:23042184

  8. Geobacter

    DEFF Research Database (Denmark)

    Lovley, Derek R; Ueki, Toshiyuki; Zhang, Tian

    2011-01-01

    of organic compounds to the reduction of insoluble Fe(III) and Mn(IV) oxides in many soils and sediments, a process of global biogeochemical significance. Some Geobacter species can anaerobically oxidize aromatic hydrocarbons and play an important role in aromatic hydrocarbon removal from contaminated...... groundwater bioremediation. The growth and activity of Geobacter species in the subsurface and their biogeochemical impact under different environmental conditions can be predicted with a systems biology approach in which genome-scale metabolic models are coupled with appropriate physical/chemical models...... are temporarily unavailable. The high conductivity of Geobacter pili and biofilms and the ability of biofilms to function as supercapacitors are novel properties that might contribute to the field of bioelectronics. The study of Geobacter species has revealed a remarkable number of microbial physiological...

  9. Genetic evidence that the degradation of para-cresol by Geobacter metallireducens is catalyzed by the periplasmic para-cresol methylhydroxylase

    DEFF Research Database (Denmark)

    Chaurasia, Akhilesh Kumar; Tremblay, Pier-Luc; Holmes, Dawn E.

    2015-01-01

    -CoA dehydrogenase (bbsG), an enzyme required for toluene degradation by G. metallireducens that is homologous to the p-hydroxybenzylsuccinyl-CoA dehydrogenase involved in p-cresol degradation by Desulfobacula toluolica Tol2 via fumarate addition, and the gene encoding the alpha prime subunit of PCMH (pcmI), were...... deleted to investigate the possibility of co-existing p-cresol degradation pathways in G. metallireducens. The absence of a functional PcmI protein completely inhibited p-cresol degradation, while deletion of the bbsG gene had little impact. These results further support the observation that G...

  10. Solution Structure of 4′-Phosphopantetheine - GmACP3 from Geobacter metallireducens: A Specialized Acyl Carrier Protein with Atypical Structural Features and a Putative Role in Lipopolysaccharide Biosynthesis†

    Science.gov (United States)

    Ramelot, Theresa A.; Smola, Matthew J.; Lee, Hsiau-Wei; Ciccosanti, Colleen; Hamilton, Keith; Acton, Thomas B.; Xiao, Rong; Everett, John K.; Prestegard, James H.; Montelione, Gaetano T.; Kennedy, Michael A.

    2011-01-01

    GmACP3 from Geobacter metallireducens is a specialized acyl carrier protein (ACP) whose gene, gmet_2339, is located near genes encoding many proteins involved in lipopolysaccharide (LPS) biosynthesis, indicating a likely function for GmACP3 in LPS production. By overexpression in Escherichia coli, about 50% holo-GmACP3 and 50% apo-GmACP3 were obtained. Apo-GmACP3 exhibited slow precipitation and non-monomeric behavior by 15N NMR relaxation measurements. Addition of 4′-phosphopantetheine (4′-PP) via enzymatic conversion by E. coli holo-ACP synthase, resulted in stable >95% holo-GmACP3 that was characterized as monomeric by 15N relaxation measurements and had no indication of conformational exchange. We have determined a high-resolution solution structure of holo-GmACP3 by standard NMR methods, including refinement with two sets of NH residual dipolar couplings, allowing for a detailed structural analysis of the interactions between 4′-PP and GmACP3. Whereas the overall four helix bundle topology is similar to previously solved ACP structures, this structure has unique characteristics, including an ordered 4′-PP conformation that places the thiol at the entrance to a central hydrophobic cavity near a conserved hydrogen-bonded Trp-His pair. These residues are part of a conserved WDSLxH/N motif found in GmACP3 and it’s orthologs. The helix locations and the large hydrophobic cavity are more similar to medium- and long-chain acyl-ACPs than to other apo- and holo-ACP structures. Taken together, structural characterization along with bioinformatic analysis of nearby genes suggest that GmACP3 is involved in lipid A acylation, possibly by atypical long-chain hydroxy fatty acids, and potentially involved in synthesis of secondary metabolites. PMID:21235239

  11. Hydrogen Production by Geobacter Species and a Mixed Consortium in a Microbial Electrolysis Cell

    KAUST Repository

    Call, D. F.; Wagner, R. C.; Logan, B. E.

    2009-01-01

    A hydrogen utilizing exoelectrogenic bacterium (Geobacter sulfurreducens) was compared to both a nonhydrogen oxidizer (Geobacter metallireducens) and a mixed consortium in order to compare the hydrogen production rates and hydrogen recoveries

  12. Comparative genomics of Geobacter chemotaxis genes reveals diverse signaling function

    Directory of Open Access Journals (Sweden)

    Antommattei Frances M

    2008-10-01

    Full Text Available Abstract Background Geobacter species are δ-Proteobacteria and are often the predominant species in a variety of sedimentary environments where Fe(III reduction is important. Their ability to remediate contaminated environments and produce electricity makes them attractive for further study. Cell motility, biofilm formation, and type IV pili all appear important for the growth of Geobacter in changing environments and for electricity production. Recent studies in other bacteria have demonstrated that signaling pathways homologous to the paradigm established for Escherichia coli chemotaxis can regulate type IV pili-dependent motility, the synthesis of flagella and type IV pili, the production of extracellular matrix material, and biofilm formation. The classification of these pathways by comparative genomics improves the ability to understand how Geobacter thrives in natural environments and better their use in microbial fuel cells. Results The genomes of G. sulfurreducens, G. metallireducens, and G. uraniireducens contain multiple (~70 homologs of chemotaxis genes arranged in several major clusters (six, seven, and seven, respectively. Unlike the single gene cluster of E. coli, the Geobacter clusters are not all located near the flagellar genes. The probable functions of some Geobacter clusters are assignable by homology to known pathways; others appear to be unique to the Geobacter sp. and contain genes of unknown function. We identified large numbers of methyl-accepting chemotaxis protein (MCP homologs that have diverse sensing domain architectures and generate a potential for sensing a great variety of environmental signals. We discuss mechanisms for class-specific segregation of the MCPs in the cell membrane, which serve to maintain pathway specificity and diminish crosstalk. Finally, the regulation of gene expression in Geobacter differs from E. coli. The sequences of predicted promoter elements suggest that the alternative sigma factors

  13. Isolation of Geobacter species from diverse sedimentary environments

    Science.gov (United States)

    Coaxes, J.D.; Phillips, E.J.P.; Lonergan, D.J.; Jenter, H.; Lovley, D.R.

    1996-01-01

    In an attempt to better understand the microorganisms responsible for Fe(III) reduction in sedimentary environments, Fe(III)-reducing microorganisms were enriched for and isolated from freshwater aquatic sediments, a pristine deep aquifer, and a petroleum-contaminated shallow aquifer. Enrichments were initiated with acetate or toluene as the electron donor and Fe(III) as the electron acceptor. Isolations were made with acetate or benzoate. Five new strains which could obtain energy for growth by dissimilatory Fe(III) reduction were isolated. All five isolates are gram- negative strict anaerobes which grow with acetate as the electron donor and Fe(III) as the electron acceptor. Analysis of the 16S rRNA sequence of the isolated organisms demonstrated that they all belonged to the genus Geobacter in the delta subdivision of the Proteobacteria. Unlike the type strain, Geobacter metallireducens, three of the five isolates could use H2 as an electron donor fur Fe(III) reduction. The deep subsurface isolate is the first Fe(III) reducer shown to completely oxidize lactate to carbon dioxide, while one of the freshwater sediment isolates is only the second Fe(III) reducer known that can oxidize toluene. The isolation of these organisms demonstrates that Geobacter species are widely distributed in a diversity of sedimentary environments in which Fe(III) reduction is an important process.

  14. Link Between Capacity for Current Production and Syntrophic Growth in Geobacter species

    Directory of Open Access Journals (Sweden)

    Amelia-Elena eRotaru

    2015-07-01

    Full Text Available Electrodes are unnatural electron acceptors, and it is yet unknown how some Geobacter species evolved to use electrodes as terminal electron acceptors. Analysis of different Geobacter species revealed that they varied in their capacity for current production. G. metallireducens and G. hydrogenophilus generated high current densities (ca. 0.05 mA/cm2, comparable to G. sulfurreducens. G. bremensis, G. chapellei, G. humireducens, and G. uranireducens, produced much lower currents (ca. 0.05 mA/cm2 and G. bemidjiensis was previously found to not produce current. There was no correspondence between the effectiveness of current generation and Fe(III oxide reduction rates. Some high-current-density strains (G. metallireducens and G. hydrogenophilus reduced Fe(III-oxides as fast as some low-current-density strains (G. bremensis, G. humireducens, and G. uranireducens whereas other low-current-density strains (G. bemidjiensis and G. chapellei reduced Fe(III oxide as slowly as G. sulfurreducens, a high-current-density strain. However, there was a correspondence between the ability to produce higher currents and the ability to grow syntrophically. G. hydrogenophilius was found to grow in co-culture with Methanosarcina barkeri, which is capable of direct interspecies electron transfer (DIET, but not with Methanospirillium hungatei capable only of H2 or formate transfer. Conductive granular activated carbon (GAC stimulated metabolism of the G. hydrogenophilus - M. barkeri co-culture, consistent with electron exchange via DIET. These findings, coupled with the previous finding that G. metallireducens and G. sulfurreducens are also capable of DIET, suggest that evolution to optimize DIET has fortuitiously conferred the capability for high-density current production to some Geobacter species.

  15. Conjugated oligoelectrolyte represses hydrogen oxidation by Geobacter sulfurreducens in microbial electrolysis cells

    KAUST Repository

    Liu, Jia

    2015-12-01

    © 2015 Elsevier B.V. A conjugated oligoelectrolyte (COE), which spontaneously aligns within cell membranes, was shown to completely inhibit H2 uptake by Geobacter sulfurreducens in microbial electrolysis cells. Coulombic efficiencies that were 490±95%, due to H2 recycling between the cathode and microorganisms on the anode, were reduced to 86±2% with COE addition. The use of the COE resulted in a 67-fold increase in H2 gas recovery, and a 4.4-fold increase in acetate removal. Current generation, H2 recovery and COD removals by Geobacter metallireducens, which cannot use H2, were unaffected by COE addition. These results show that this COE is an effective H2 uptake inhibitor, and that it can enable improved and sustained H2 gas recovery in this bioelectrochemical system.

  16. Hydrogen Production by Geobacter Species and a Mixed Consortium in a Microbial Electrolysis Cell

    KAUST Repository

    Call, D. F.

    2009-10-09

    A hydrogen utilizing exoelectrogenic bacterium (Geobacter sulfurreducens) was compared to both a nonhydrogen oxidizer (Geobacter metallireducens) and a mixed consortium in order to compare the hydrogen production rates and hydrogen recoveries of pure and mixed cultures in microbial electrolysis cells (MECs). At an applied voltage of 0.7 V, both G. sulfurreducens and the mixed culture generated similar current densities (ca. 160 A/m3), resulting in hydrogen production rates of ca. 1.9 m3 H2/m 3/day, whereas G. metallireducens exhibited lower current densities and production rates of 110 ± 7 A/m3 and 1.3 ± 0.1 m3 H2/m3/day, respectively. Before methane was detected in the mixed-culture MEC, the mixed consortium achieved the highest overall energy recovery (relative to both electricity and substrate energy inputs) of 82% ± 8% compared to G. sulfurreducens (77% ± 2%) and G. metallireducens (78% ± 5%), due to the higher coulombic efficiency of the mixed consortium. At an applied voltage of 0.4 V, methane production increased in the mixed-culture MEC and, as a result, the hydrogen recovery decreased and the overall energy recovery dropped to 38% ± 16% compared to 80% ± 5% for G. sulfurreducens and 76% ± 0% for G. metallireducens. Internal hydrogen recycling was confirmed since the mixed culture generated a stable current density of 31 ± 0 A/m3 when fed hydrogen gas, whereas G. sulfurreducens exhibited a steady decrease in current production. Community analysis suggested that G. sulfurreducens was predominant in the mixed-culture MEC (72% of clones) despite its relative absence in the mixed-culture inoculum obtained from a microbial fuel cell reactor (2% of clones). These results demonstrate that Geobacter species are capable of obtaining similar hydrogen production rates and energy recoveries as mixed cultures in an MEC and that high coulombic efficiencies in mixed culture MECs can be attributed in part to the recycling of hydrogen into current. Copyright

  17. Hydrogen Production by Geobacter Species and a Mixed Consortium in a Microbial Electrolysis Cell▿

    Science.gov (United States)

    Call, Douglas F.; Wagner, Rachel C.; Logan, Bruce E.

    2009-01-01

    A hydrogen utilizing exoelectrogenic bacterium (Geobacter sulfurreducens) was compared to both a nonhydrogen oxidizer (Geobacter metallireducens) and a mixed consortium in order to compare the hydrogen production rates and hydrogen recoveries of pure and mixed cultures in microbial electrolysis cells (MECs). At an applied voltage of 0.7 V, both G. sulfurreducens and the mixed culture generated similar current densities (ca. 160 A/m3), resulting in hydrogen production rates of ca. 1.9 m3 H2/m3/day, whereas G. metallireducens exhibited lower current densities and production rates of 110 ± 7 A/m3 and 1.3 ± 0.1 m3 H2/m3/day, respectively. Before methane was detected in the mixed-culture MEC, the mixed consortium achieved the highest overall energy recovery (relative to both electricity and substrate energy inputs) of 82% ± 8% compared to G. sulfurreducens (77% ± 2%) and G. metallireducens (78% ± 5%), due to the higher coulombic efficiency of the mixed consortium. At an applied voltage of 0.4 V, methane production increased in the mixed-culture MEC and, as a result, the hydrogen recovery decreased and the overall energy recovery dropped to 38% ± 16% compared to 80% ± 5% for G. sulfurreducens and 76% ± 0% for G. metallireducens. Internal hydrogen recycling was confirmed since the mixed culture generated a stable current density of 31 ± 0 A/m3 when fed hydrogen gas, whereas G. sulfurreducens exhibited a steady decrease in current production. Community analysis suggested that G. sulfurreducens was predominant in the mixed-culture MEC (72% of clones) despite its relative absence in the mixed-culture inoculum obtained from a microbial fuel cell reactor (2% of clones). These results demonstrate that Geobacter species are capable of obtaining similar hydrogen production rates and energy recoveries as mixed cultures in an MEC and that high coulombic efficiencies in mixed culture MECs can be attributed in part to the recycling of hydrogen into current. PMID:19820150

  18. Carboxydotrophic growth of Geobacter sulfurreducens

    NARCIS (Netherlands)

    Geelhoed, J.S.; Henstra, A.M.; Stams, A.J.M.

    2016-01-01

    This study shows that Geobacter sulfurreducens grows on carbon monoxide (CO) as electron donor with fumarate as electron acceptor. Geobacter sulfurreducens was tolerant to high CO levels, with up to 150 kPa in the headspace tested. During growth, hydrogen was detected in very slight amounts (∼5

  19. Orenia metallireducens sp. nov. Strain Z6, a Novel Metal-Reducing Member of the Phylum Firmicutes from the Deep Subsurface

    Science.gov (United States)

    Sanford, Robert A.; Boyanov, Maxim I.; Kemner, Kenneth M.; O'Loughlin, Edward J.; Chang, Yun-juan; Locke, Randall A.; Weber, Joseph R.; Egan, Sheila M.; Mackie, Roderick I.; Cann, Isaac; Fouke, Bruce W.

    2016-01-01

    ABSTRACT A novel halophilic and metal-reducing bacterium, Orenia metallireducens strain Z6, was isolated from briny groundwater extracted from a 2.02 km-deep borehole in the Illinois Basin, IL. This organism shared 96% 16S rRNA gene similarity with Orenia marismortui but demonstrated physiological properties previously unknown for this genus. In addition to exhibiting a fermentative metabolism typical of the genus Orenia, strain Z6 reduces various metal oxides [Fe(III), Mn(IV), Co(III), and Cr(VI)], using H2 as the electron donor. Strain Z6 actively reduced ferrihydrite over broad ranges of pH (6 to 9.6), salinity (0.4 to 3.5 M NaCl), and temperature (20 to 60°C). At pH 6.5, strain Z6 also reduced more crystalline iron oxides, such as lepidocrocite (γ-FeOOH), goethite (α-FeOOH), and hematite (α-Fe2O3). Analysis of X-ray absorption fine structure (XAFS) following Fe(III) reduction by strain Z6 revealed spectra from ferrous secondary mineral phases consistent with the precipitation of vivianite [Fe3(PO4)2] and siderite (FeCO3). The draft genome assembled for strain Z6 is 3.47 Mb in size and contains 3,269 protein-coding genes. Unlike the well-understood iron-reducing Shewanella and Geobacter species, this organism lacks the c-type cytochromes for typical Fe(III) reduction. Strain Z6 represents the first bacterial species in the genus Orenia (order Halanaerobiales) reported to reduce ferric iron minerals and other metal oxides. This microbe expands both the phylogenetic and physiological scopes of iron-reducing microorganisms known to inhabit the deep subsurface and suggests new mechanisms for microbial iron reduction. These distinctions from other Orenia spp. support the designation of strain Z6 as a new species, Orenia metallireducens sp. nov. IMPORTANCE A novel iron-reducing species, Orenia metallireducens sp. nov., strain Z6, was isolated from groundwater collected from a geological formation located 2.02 km below land surface in the Illinois Basin, USA

  20. Mechanisms for Electron Transfer Through Pili to Fe(III) Oxide in Geobacter

    Energy Technology Data Exchange (ETDEWEB)

    Lovley, Derek R. [Univ. of Massachusetts, Amherst, MA (United States)

    2015-03-09

    The purpose of these studies was to aid the Department of Energy in its goal of understanding how microorganisms involved in the bioremediation of metals and radionuclides sustain their activity in the subsurface. This information is required in order to incorporate biological processes into decision making for environmental remediation and long-term stewardship of contaminated sites. The proposed research was designed to elucidate the mechanisms for electron transfer to Fe(III) oxides in Geobacter species because Geobacter species are abundant dissimilatory metal-reducing microorganisms in a diversity of sites in which uranium is undergoing natural attenuation via the reduction of soluble U(VI) to insoluble U(IV) or when this process is artificially stimulated with the addition of organic electron donors. This study investigated the novel, but highly controversial, concept that the final conduit for electron transfer to Fe(III) oxides are electrically conductive pili. The specific objectives were to: 1) further evaluate the conductivity along the pili of Geobacter sulfurreducens and related organisms; 2) determine the mechanisms for pili conductivity; and 3) investigate the role of pili in Fe(III) oxide reduction. The studies demonstrated that the pili of G. sulfurreducens are conductive along their length. Surprisingly, the pili possess a metallic-like conductivity similar to that observed in synthetic organic conducting polymers such as polyaniline. Detailed physical analysis of the pili, as well as studies in which the structure of the pili was genetically modified, demonstrated that the metallic-like conductivity of the pili could be attributed to overlapping pi-pi orbitals of aromatic amino acids. Other potential mechanisms for conductivity, such as electron hopping between cytochromes associated with the pili were definitively ruled out. Pili were also found to be essential for Fe(III) oxide reduction in G. metallireducens. Ecological studies demonstrated

  1. Identification of geobacter populations in the uranium mill tailings Shiprock

    International Nuclear Information System (INIS)

    Radeva, G.; Selenska-Pobell, S.

    2006-01-01

    Geobacter - specific primers were used for construction of a 16S rDNA library for a water sample collected from uranium mill tailings near Shiprock (Sh853) in the USA . Most of the retrieved sequences were affiliated with different Geobacter species, however sequences related to other δ -Proteobacteria were identified as well. (authors)

  2. Proteogenomic monitoring of Geobacter physiology during stimulated uranium bioremediation

    Energy Technology Data Exchange (ETDEWEB)

    Wilkins, M.J.; VerBerkmoes, N.C.; Williams, K.H.; Callister, S.J.; Mouser, P.J.; Elifantz, H.; N' Guessan, A.L.; Thomas, B.C.; Nicora, C.D.; Shah, M.B.; Lipton, M.S.; Lovley, D.R.; Hettich, R.L.; Long, P.E.; Banfield, J.F.; Abraham, P.

    2009-08-01

    Implementation of uranium bioremediation requires methods for monitoring the membership and activities of the subsurface microbial communities that are responsible for reduction of soluble U(VI) to insoluble U(IV). Here, we report a proteomics-based approach for simultaneously documenting the strain membership and microbial physiology of the dominant Geobacter community members during in situ acetate amendment of the U-contaminated Rifle, CO, aquifer. Three planktonic Geobacter-dominated samples were obtained from two wells down-gradient of acetate addition. Over 2,500 proteins from each of these samples were identified by matching liquid chromatography-tandem mass spectrometry spectra to peptides predicted from seven isolate Geobacter genomes. Genome-specific peptides indicate early proliferation of multiple M21 and Geobacter bemidjiensis-like strains and later possible emergence of M21 and G. bemidjiensis-like strains more closely related to Geobacter lovleyi. Throughout biostimulation, the proteome is dominated by enzymes that convert acetate to acetyl-coenzyme A and pyruvate for central metabolism, while abundant peptides matching tricarboxylic acid cycle proteins and ATP synthase subunits were also detected, indicating the importance of energy generation during the period of rapid growth following the start of biostimulation. Evolving Geobacter strain composition may be linked to changes in protein abundance over the course of biostimulation and may reflect changes in metabolic functioning. Thus, metagenomics-independent community proteogenomics can be used to diagnose the status of the subsurface consortia upon which remediation biotechnology relies.

  3. Extracellular Palladium Nanoparticle Production using Geobacter sulfurreducens

    KAUST Repository

    Yates, Matthew D.

    2013-09-03

    Sustainable methods are needed to recycle precious metals and synthesize catalytic nanoparticles. Palladium nanoparticles can be produced via microbial reduction of soluble Pd(II) to Pd(0), but in previous tests using dissimilatory metal reducing bacteria (DMRB), the nanoparticles were closely associated with the cells, occupying potential reductive sites and eliminating the potential for cell reuse. The DMRB Geobacter sulfurreducens was shown here to reduce soluble Pd(II) to Pd(0) nanoparticles primarily outside the cell, reducing the toxicity of metal ions, and allowing nanoparticle recovery without cell destruction that has previously been observed using other microorganisms. Cultures reduced 50 ± 3 mg/L Pd(II) with 1% hydrogen gas (v/v headspace) in 6 h incubation tests [100 mg/L Pd(II) initially], compared to 8 ± 3 mg/L (10 mM acetate) without H2. Acetate was ineffective as an electron donor for palladium removal in the presence or absence of fumarate as an electron acceptor. TEM imaging verified that Pd(0) nanoparticles were predominantly in the EPS surrounding cells in H2-fed cultures, with only a small number of particles visible inside the cell. Separation of the cells and EPS by centrifugation allowed reuse of the cell suspensions and effective nanoparticle recovery. These results demonstrate effective palladium recovery and nanoparticle production using G. sulfurreducens cell suspensions and renewable substrates such as H2 gas. © 2013 American Chemical Society.

  4. Geobacter anodireducens sp. nov., an exoelectrogenic microbe in bioelectrochemical systems

    KAUST Repository

    Sun, D.

    2014-07-22

    © 2014 IUMS. A previously isolated exoelectrogenic bacterium, strain SD-1(T), was further characterized and identified as a representative of a novel species of the genus Geobacter. Strain SD-1(T) was Gram-negative, aerotolerant, anaerobic, non-spore-forming, non-fermentative and non-motile. Cells were short, curved rods (0.8-1.3 µm long and 0.3 µm in diameter). Growth of strain SD-1(T) was observed at 15-42 °C and pH 6.0-8.5, with optimal growth at 30-35 °C and pH 7. Analysis of 16S rRNA gene sequences indicated that the isolate was a member of the genus Geobacter, with the closest known relative being Geobacter sulfurreducens PCA(T) (98% similarity). Similar to other members of the genus Geobacter, strain SD-1(T) used soluble or insoluble Fe(III) as the sole electron acceptor coupled with the oxidation of acetate. However, SD-1(T) could not reduce fumarate as an electron acceptor with acetate oxidization, which is an important physiological trait for G. sulfurreducens. Moreover, SD-1(T) could grow in media containing as much as 3% NaCl, while G. sulfurreducens PCA(T) can tolerate just half this concentration, and this difference in salt tolerance was even more obvious when cultivated in bioelectrochemical systems. DNA-DNA hybridization analysis of strain SD-1(T) and its closest relative, G. sulfurreducens ATCC 51573(T), showed a relatedness of 61.6%. The DNA G+C content of strain SD-1(T) was 58.9 mol%. Thus, on the basis of these characteristics, strain SD-1(T) was not assigned to G. sulfurreducens, and was instead classified in the genus Geobacter as a representative of a novel species. The name Geobacter anodireducens sp. nov. is proposed, with the type strain SD-1(T) ( = CGMCC 1.12536(T) = KCTC 4672(T)).

  5. Electron shuttle-mediated biotransformation of hexahydro-1,3,5-trinitro-1,3,5-triazine adsorbed to granular activated carbon.

    Science.gov (United States)

    Millerick, Kayleigh; Drew, Scott R; Finneran, Kevin T

    2013-08-06

    Granular activated carbon (GAC) effectively removes hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) from groundwater but generates RDX-laden GAC that must be disposed of or regenerated. Batch reactors containing GAC to which RDX was preadsorbed were used in experiments to test the potential for adsorbed RDX reduction and daughter product formation using (i) chemically reduced anthrahydroquinone-2,6-disulfonate (AH2QDS), (ii) resting Geobacter metallireducens strain GS-15, and (iii) a combined system containing AQDS and GS-15. Approximately 97.0% of the adsorbed RDX was transformed in each of these experimental systems by 90 h. Chemically reduced AQDS (AH2QDS) transformed 99.2% of adsorbed RDX; formaldehyde was produced rapidly and was stoichiometric (3 mol HCHO per mol RDX). Geobacter metallireducens also reduced RDX with and without AQDS present. This is the first study to demonstrate biological transformation of RDX adsorbed to GAC. Formaldehyde increased and then decreased in biological systems, suggesting a previously unreported capacity for G. metallireducens to oxidize formaldehyde, which was confirmed with resting cell suspensions. These data suggest the masses of GAC waste currently produced by activated carbon at RDX remediation sites can be minimized, decreasing the carbon footprint of the treatment technology. Alternatively, this strategy may be used to develop a Bio-GAC system for ex situ RDX treatment.

  6. Enhanced Uranium Immobilization and Reduction by Geobacter sulfurreducens Biofilms

    Science.gov (United States)

    Cologgi, Dena L.; Speers, Allison M.; Bullard, Blair A.; Kelly, Shelly D.

    2014-01-01

    Biofilms formed by dissimilatory metal reducers are of interest to develop permeable biobarriers for the immobilization of soluble contaminants such as uranium. Here we show that biofilms of the model uranium-reducing bacterium Geobacter sulfurreducens immobilized substantially more U(VI) than planktonic cells and did so for longer periods of time, reductively precipitating it to a mononuclear U(IV) phase involving carbon ligands. The biofilms also tolerated high and otherwise toxic concentrations (up to 5 mM) of uranium, consistent with a respiratory strategy that also protected the cells from uranium toxicity. The enhanced ability of the biofilms to immobilize uranium correlated only partially with the biofilm biomass and thickness and depended greatly on the area of the biofilm exposed to the soluble contaminant. In contrast, uranium reduction depended on the expression of Geobacter conductive pili and, to a lesser extent, on the presence of the c cytochrome OmcZ in the biofilm matrix. The results support a model in which the electroactive biofilm matrix immobilizes and reduces the uranium in the top stratum. This mechanism prevents the permeation and mineralization of uranium in the cell envelope, thereby preserving essential cellular functions and enhancing the catalytic capacity of Geobacter cells to reduce uranium. Hence, the biofilms provide cells with a physically and chemically protected environment for the sustained immobilization and reduction of uranium that is of interest for the development of improved strategies for the in situ bioremediation of environments impacted by uranium contamination. PMID:25128347

  7. Microbial electrocatalysis with Geobacter sulfurreducens biofilm on stainless steel cathodes

    OpenAIRE

    Dumas, Claire; Basséguy, Régine; Bergel, Alain

    2008-01-01

    Stainless steel and graphite electrodes were individually addressed and polarized at−0.60V vs. Ag/AgCl in reactors filled with a growth medium that contained 25mM fumarate as the electron acceptor and no electron donor, in order to force the microbial cells to use the electrode as electron source. When the reactor was inoculated with Geobacter sulfurreducens, the current increased and stabilized at average values around 0.75Am−2 for graphite and 20.5Am−2 for stainless steel. Cyclic voltamm...

  8. The Complex Conductivity Signature of Geobacter Species in Geological Media

    Science.gov (United States)

    Brown, I.; Atekwana, E. A.; Sarkisova, S.; Achang, M.

    2013-12-01

    The Complex Conductivity (CC) technique is a promising biogeophysical approach for sensing microbially-induced changes in geological media because of its low-invasive character and sufficient sensitivity to enhanced microbial activity in the near subsurface. Geobacter species have been shown to play important roles in the bioremediation of groundwater contaminated with petroleum and landfill leachate. This capability is based on the ability of Geobacter species to reduce Fe(III) by transferring of electrons from the reduced equivalents to Fe(III) rich minerals through respiration chain and special metallic-like conductors - pili. Only the cultivation of Geobacter species on Fe(III) oxides specifically express pili biosynthesis. Moreover, mutants that cannot produce pili are unable to reduce Fe(III) oxides. However, little is known about the contribution of these molecular conductors (nanowires) to the generation of complex conductivity signatures in geological media. Here, we present the results about the modulation of CC signatures in geological media by Geobacter sulfurreducens (G.s.). Cultures of wild strain G.s. and its pilA(-) mutant were anaerobically cultivated in the presence of the pair of such donors and acceptors of electrons: acetate - fumarate, and acetate - magnetite under anaerobic conditions. Each culture was injected in CC sample holders filled either with N2-CO2 mix (planktonic variant) or with this gases mix and glass beads, d=1 mm, (porous medium variant). Both strains of G.s. proliferated well in a medium supplemented with acetate-fumarate. However, pilA(-) mutant did not multiply in a medium supplemented with ox-red pair yeast extract - magnetite. This observation confirmed that only wild pilA(+) strain is capable of the dissimilatory reduction of Fe(III) within magnetite molecule. The measurement of CC responses from planktonic culture of G.s. wild strain grown with acetate-fumarate did not show linear correlation with their magnitudes but

  9. Use of a Coculture To Enable Current Production by Geobacter sulfurreducens

    KAUST Repository

    Qu, Y.; Feng, Y.; Wang, X.; Logan, B. E.

    2012-01-01

    Microbial fuel cells often produce more electrical power with mixed cultures than with pure cultures. Here, we show that a coculture of a nonexoelectrogen (Escherichia coli) and Geobacter sulfurreducens improved system performance relative

  10. Conjugated oligoelectrolyte represses hydrogen oxidation by Geobacter sulfurreducens in microbial electrolysis cells

    KAUST Repository

    Liu, Jia; Hou, Huijie; Chen, Xiaofen; Bazan, Guillermo C.; Kashima, Hiroyuki; Logan, Bruce

    2015-01-01

    © 2015 Elsevier B.V. A conjugated oligoelectrolyte (COE), which spontaneously aligns within cell membranes, was shown to completely inhibit H2 uptake by Geobacter sulfurreducens in microbial electrolysis cells. Coulombic efficiencies that were 490

  11. Conductive particles enable syntrophic acetate oxidation between Geobacter and Methanosarcina from coastal sediments

    DEFF Research Database (Denmark)

    Rotaru, Amelia-Elena; Calabrese, Federica; Stryhanyuk, Hryhoriy

    2017-01-01

    pressure and their survival depends on successful partnership. Here we demonstrate that conductive minerals facilitate a SAO partnership between Geobacter and Methanosarcina from the coastal sediments of the Bothnian Bay, Baltic Sea. Bothnian methanogenic sediments showed a high apparent isotopic...... fractionation (αc 1.07) characteristic of CO2-reductive methanogenesis. The native community was represented by electrogens such as Geobacter and methanogens like Methanosarcina. Upon the addition of conductive particles (activated carbon and magnetite), methanogenesis from acetate increased fourfold. Geobacter...... (96% related to G. psychrophilus) and Methanosarcina (99% related to M. subterranea) dominated the conductive particle-spiked SAO communities. Using NanoSIMS we demonstrated that during SAO, Geobacter incorporated 82% of the labeled acetate as compared to only 18% by Methanosarcina. At the same time...

  12. Integrative analysis of Geobacter spp. and sulfate-reducing bacteria during uranium bioremediation

    Directory of Open Access Journals (Sweden)

    D. Lovley

    2012-03-01

    Full Text Available Enhancing microbial U(VI reduction with the addition of organic electron donors is a promising strategy for immobilizing uranium in contaminated groundwaters, but has yet to be optimized because of a poor understanding of the factors controlling the growth of various microbial communities during bioremediation. In previous field trials in which acetate was added to the subsurface, there were two distinct phases: an initial phase in which acetate-oxidizing, U(VI-reducing Geobacter predominated and U(VI was effectively reduced and a second phase in which acetate-oxidizing sulfate reducing bacteria (SRB predominated and U(VI reduction was poor. The interaction of Geobacter and SRB was investigated both in sediment incubations that mimicked in situ bioremediation and with in silico metabolic modeling. In sediment incubations, Geobacter grew quickly but then declined in numbers as the microbially reducible Fe(III was depleted whereas the SRB grow more slowly and reached dominance after 30–40 days. Modeling predicted a similar outcome. Additional modeling in which the relative initial percentages of the Geobacter and SRB were varied indicated that there was little to no competitive interaction between Geobacter and SRB when acetate was abundant. Further simulations suggested that the addition of Fe(III would revive the Geobacter, but have little to no effect on the SRB. This result was confirmed experimentally. The results demonstrate that it is possible to predict the impact of amendments on important components of the subsurface microbial community during groundwater bioremediation. The finding that Fe(III availability, rather than competition with SRB, is the key factor limiting the activity of Geobacter during in situ uranium bioremediation will aid in the design of improved uranium bioremediation strategies.

  13. Microbial electrocatalysis with Geobacter sulfurreducens biofilm on stainless steel cathodes

    International Nuclear Information System (INIS)

    Dumas, Claire; Basseguy, Regine; Bergel, Alain

    2008-01-01

    Stainless steel and graphite electrodes were individually addressed and polarized at -0.60 V vs. Ag/AgCl in reactors filled with a growth medium that contained 25 mM fumarate as the electron acceptor and no electron donor, in order to force the microbial cells to use the electrode as electron source. When the reactor was inoculated with Geobacter sulfurreducens, the current increased and stabilized at average values around 0.75 A m -2 for graphite and 20.5 A m -2 for stainless steel. Cyclic voltammetry performed at the end of the experiment indicated that the reduction started at around -0.30 V vs. Ag/AgCl on stainless steel. Removing the biofilm formed on the electrode surface made the current totally disappear, confirming that the G.sulfurreducens biofilm was fully responsible for the electrocatalysis of fumarate reduction. Similar current densities were recorded when the electrodes were polarized after being kept in open circuit for several days. The reasons for the bacteria presence and survival on non-connected stainless steel coupons were discussed. Chronoamperometry experiments performed at different potential values suggested that the biofilm-driven catalysis was controlled by electrochemical kinetics. The high current density obtained, quite close to the redox potential of the fumarate/succinate couple, presents stainless steel as a remarkable material to support biocathodes

  14. G eobacter sp. SD-1 with enhanced electrochemical activity in high-salt concentration solutions

    KAUST Repository

    Sun, Dan; Call, Douglas; Wang, Aijie; Cheng, Shaoan; Logan, Bruce E.

    2014-01-01

    © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd. Summary: An isolate, designated strain SD-1, was obtained from a biofilm dominated by Geobacter sulfurreducens in a microbial fuel cell. The electrochemical activity of strain SD-1 was compared with type strains, G.sulfurreducensPCA and Geobacter metallireducensGS-15, and a mixed culture in microbial electrolysis cells. SD-1 produced a maximum current density of 290±29Am-3 in a high-concentration phosphate buffer solution (PBS-H, 200mM). This current density was significantly higher than that produced by the mixed culture (189±44Am-3) or the type strains (<70Am-3). In a highly saline water (SW; 50mM PBS and 650mM NaCl), current by SD-1 (158±4Am-3) was reduced by 28% compared with 50mM PBS (220±4Am-3), but it was still higher than that of the mixed culture (147±19Am-3), and strains PCA and GS-15 did not produce any current. Electrochemical tests showed that the improved performance of SD-1 was due to its lower charge transfer resistance and more negative potentials produced at higher current densities. These results show that the electrochemical activity of SD-1 was significantly different than other Geobacter strains and mixed cultures in terms of its salt tolerance.

  15. G eobacter sp. SD-1 with enhanced electrochemical activity in high-salt concentration solutions

    KAUST Repository

    Sun, Dan

    2014-07-16

    © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd. Summary: An isolate, designated strain SD-1, was obtained from a biofilm dominated by Geobacter sulfurreducens in a microbial fuel cell. The electrochemical activity of strain SD-1 was compared with type strains, G.sulfurreducensPCA and Geobacter metallireducensGS-15, and a mixed culture in microbial electrolysis cells. SD-1 produced a maximum current density of 290±29Am-3 in a high-concentration phosphate buffer solution (PBS-H, 200mM). This current density was significantly higher than that produced by the mixed culture (189±44Am-3) or the type strains (<70Am-3). In a highly saline water (SW; 50mM PBS and 650mM NaCl), current by SD-1 (158±4Am-3) was reduced by 28% compared with 50mM PBS (220±4Am-3), but it was still higher than that of the mixed culture (147±19Am-3), and strains PCA and GS-15 did not produce any current. Electrochemical tests showed that the improved performance of SD-1 was due to its lower charge transfer resistance and more negative potentials produced at higher current densities. These results show that the electrochemical activity of SD-1 was significantly different than other Geobacter strains and mixed cultures in terms of its salt tolerance.

  16. Rational engineering of Geobacter sulfurreducens electron transfer components: a foundation for building improved Geobacter-based bioelectrochemical technologies

    Directory of Open Access Journals (Sweden)

    Joana M Dantas

    2015-07-01

    Full Text Available Multiheme cytochromes have been implicated in Geobacter sulfurreducens (Gs extracellular electron transfer (EET. These proteins are potential targets to improve EET and enhance bioremediation and electrical current production by Gs. However, the functional characterization of multiheme cytochromes is particularly complex due to the co-existence of several microstates in solution, connecting the fully reduced and fully oxidized states. Over the last decade, new strategies have been developed to characterize multiheme redox proteins functionally and structurally. These strategies were used to reveal the functional mechanism of Gs multiheme cytochromes and also to identify key residues in these proteins for EET. In previous studies, we set the foundations for enhancement of the EET abilities of Gs by characterizing a family of five triheme cytochromes (PpcA-E. These periplasmic cytochromes are implicated in electron transfer between the oxidative reactions of metabolism in the cytoplasm and the reduction of extracellular terminal electron acceptors at the cell’s outer surface. The results obtained suggested that PpcA can couple e-/H+ transfer, a property that might contribute to the proton electrochemical gradient across the cytoplasmic membrane for metabolic energy production. The structural and functional properties of PpcA were characterized in detail and used for rational design of a family of 23 single site PpcA mutants. In this review, we summarize the functional characterization of the native and mutant proteins. Mutants that retain the mechanistic features of PpcA and adopt preferential e-/H+ transfer pathways at lower reduction potential values compared to the wild-type protein were selected for in vivo studies as the best candidates to increase the electron transfer rate of Gs. For the first time Gs strains have been manipulated by the introduction of mutant forms of essential proteins with the aim to develop and improve

  17. Mechanisms for the reduction of actinide ions by Geobacter sulfurreducens

    International Nuclear Information System (INIS)

    Renshaw, J.C.; Livens, F.R.; May, I.; Lloyd, J.R.

    2005-01-01

    Full text of publication follows: Three of the most problematic radioactive contaminants are the actinide elements uranium, neptunium and plutonium. All three pose considerable long-term environmental risks. The most stable environmental oxidation states of uranium and neptunium are VI and V, respectively, as the di-oxo cations [UO 2 ] 2+ and [NpO 2 ] + ; both are highly soluble and so are relatively mobile and biologically available in the environment. In similar conditions, plutonium mainly exists as Pu(IV), which forms a highly insoluble hydrous oxide, although is also environmentally stable in the more soluble III, V and VI oxidation states. The bio-reduction of U(VI) by anaerobic subsurface microorganisms has been the focus of much recent interest. Both Fe(III)- and sulfate-reducing bacteria have been shown to reduce soluble [U VI O 2 ] 2+ to insoluble U IV O 2 , with c-type cytochromes involved in electron transfer to the actinide. Such transformations offer a strategy for the bio-remediation of uranium contaminated groundwater and a potential mechanism for the bio-deposition of uranium ores. The mechanism of U(VI). reduction has important implications for the potential microbial reduction of transuranic elements with environmentally stable lower oxidation states. Reduction of mobile 237 Np(V) to Np(IV) and subsequent precipitation may be advantageous whilst remobilization of immobile Pu(IV) as more soluble Pu(III) species could have important environmental implications. Conversely, selective reduction might allow targeting of particular radionuclide species. The model anaerobic bacterium Geobacter sulfurreducens is typical of those found in contaminated subsurface environments and has been shown to reduce soluble [U VI O 2 ] 2+ to insoluble U IV O 2 . In the course of this study we use X-ray absorption spectroscopy (XAS) to show that G. sulfurreducens reduces U(VI) by a one-electron reduction, forming an unstable [UO 2 ] + species which subsequently

  18. High Biofilm Conductivity Maintained Despite Anode Potential Changes in a Geobacter-Enriched Biofilm

    Science.gov (United States)

    This study systematically assessed intracellular electron transfer (IET) and extracellular electron transfer (EET) kinetics with respect to anode potential (Eanode) in a mixed-culture biofilm anode enriched with Geobacter spp. High biofilm conductivity (0.96–1.24 mScm^-1) was mai...

  19. Electricity-Assisted Biological Hydrogen Production from Acetate by Geobacter sulfurreducens

    NARCIS (Netherlands)

    Geelhoed, J.S.; Stams, A.J.M.

    2011-01-01

    Geobacter sulfurreducens is a well-known current-producing microorganism in microbial fuel cells, and is able to use acetate and hydrogen as electron donor. We studied the functionality of G. sulfurreducens as biocatalyst for hydrogen formation at the cathode of a microbial electrolysis cell (MEC).

  20. Use of a Coculture To Enable Current Production by Geobacter sulfurreducens

    KAUST Repository

    Qu, Y.

    2012-02-17

    Microbial fuel cells often produce more electrical power with mixed cultures than with pure cultures. Here, we show that a coculture of a nonexoelectrogen (Escherichia coli) and Geobacter sulfurreducens improved system performance relative to that of a pure culture of the exoelectrogen due to the consumption of oxygen leaking into the reactor.

  1. Maintenance of Geobacter-dominated biofilms in microbial fuel cells treating synthetic wastewater.

    Science.gov (United States)

    Commault, Audrey S; Lear, Gavin; Weld, Richard J

    2015-12-01

    Geobacter-dominated biofilms can be selected under stringent conditions that limit the growth of competing bacteria. However, in many practical applications, such stringent conditions cannot be maintained and the efficacy and stability of these artificial biofilms may be challenged. In this work, biofilms were selected on low-potential anodes (-0.36 V vs Ag/AgCl, i.e. -0.08 V vs SHE) in minimal acetate or ethanol media. Selection conditions were then relaxed by transferring the biofilms to synthetic wastewater supplemented with soil as a source of competing bacteria. We tracked community succession and functional changes in these biofilms. The Geobacter-dominated biofilms showed stability in their community composition and electrochemical properties, with Geobacter sp. being still electrically active after six weeks in synthetic wastewater with power densities of 100±19 mW·m(-2) (against 74±14 mW·m(-2) at week 0) for all treatments. After six weeks, the ethanol-selected biofilms, despite their high taxon richness and their efficiency at removing the chemical oxygen demand (0.8 g·L(-1) removed against the initial 1.3 g·L(-1) injected), were the least stable in terms of community structure. These findings have important implications for environmental microbial fuel cells based on Geobacter-dominated biofilms and suggest that they could be stable in challenging environments. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Flux analysis of central metabolic pathways in Geobactermetallireducens during reduction of solubleFe(III)-NTA

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Yinjie J.; Chakraborty, Romy; Garcia-Martin, Hector; Chu,Jeannie; Hazen, Terry C.; Keasling, Jay D.

    2007-01-01

    We analyzed the carbon fluxes in the central metabolism ofGeobacter metallireducens strain GS-15 using 13C isotopomer modeling.Acetate labeled in the 1st or 2nd position was the sole carbon source,and Fe-NTA was the sole terminal electron acceptor. The measured labeledacetate uptake rate was 21 mmol/gdw/h in the exponential growth phase.The resulting isotope labeling pattern of amino acids allowed an accuratedetermination of the in vivo global metabolic reaction rates (fluxes)through the central metabolic pathways using a computational isotopomermodel. The tracer experiments showed that G. metallireducens containedcomplete biosynthesis pathways for essential metabolism, and this strainmight also have an unusual isoleucine biosynthesis route (usingacetyl-CoA and pyruvate as the precursors). The model indicated that over90 percent of the acetate was completely oxidized to CO2 via a completetricarboxylic acid (TCA) cycle while reducing iron. Pyruvate carboxylaseand phosphoenolpyruvate carboxykinase were present under theseconditions, but enzymes in the glyoxylate shunt and malic enzyme wereabsent. Gluconeogenesis and the pentose phosphate pathway were mainlyemployed for biosynthesis and accounted for less than 3 percent of totalcarbon consumption. The model also indicated surprisingly highreversibility in the reaction between oxoglutarate and succinate. Thisstep operates close to the thermodynamic equilibrium possibly becausesuccinate is synthesized via a transferase reaction, and the conversionof oxoglutarate to succinate is a rate limiting step for carbonmetabolism. These findings enable a better understanding of therelationship between genome annotation and extant metabolic pathways inG. metallireducens.

  3. Lactate Oxidation Coupled to Iron or Electrode Reduction by Geobacter sulfurreducens PCA

    KAUST Repository

    Call, D. F.

    2011-10-14

    Geobacter sulfurreducens PCA completely oxidized lactate and reduced iron or an electrode, producing pyruvate and acetate intermediates. Compared to the current produced by Shewanella oneidensis MR-1, G. sulfurreducens PCA produced 10-times-higher current levels in lactate-fed microbial electrolysis cells. The kinetic and comparative analyses reported here suggest a prominent role of G. sulfurreducens strains in metaland electrode-reducing communities supplied with lactate. © 2011, American Society for Microbiology.

  4. Lactate Oxidation Coupled to Iron or Electrode Reduction by Geobacter sulfurreducens PCA

    KAUST Repository

    Call, D. F.; Logan, B. E.

    2011-01-01

    Geobacter sulfurreducens PCA completely oxidized lactate and reduced iron or an electrode, producing pyruvate and acetate intermediates. Compared to the current produced by Shewanella oneidensis MR-1, G. sulfurreducens PCA produced 10-times-higher current levels in lactate-fed microbial electrolysis cells. The kinetic and comparative analyses reported here suggest a prominent role of G. sulfurreducens strains in metaland electrode-reducing communities supplied with lactate. © 2011, American Society for Microbiology.

  5. Dicty_cDB: Contig-U16581-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 915_913( CP000915 |pid:none) Francisella tularensis subsp. me... 174 1e-41 AP008232_642( AP008232 |pid:none) Sodalis glossini...( FM992690 |pid:none) Candida dubliniensis CD36 chromo... 227 9e-58 (Q9Y7F0) RecName: Full=Peroxiredoxin TSA...9( CP000148 |pid:none) Geobacter metallireducens GS-15... 199 3e-49 AY842247_1( AY842247 |pid:none) Leishmania amazonensi...2e-48 AE001363_758( AE001363 |pid:none) Chlamydophila pneumoniae CWL029,... 196 2e-48 CP001071_1298( CP001071 |pid:none) Akkermansi...none) Helicobacter pylori isolate HP52 a... 169 3e-40 CU468135_2542( CU468135 |pid:none) Erwinia tasmaniensi

  6. Enrichment of Geobacter species in response to stimulation of Fe(III) reduction in sandy aquifer sediments

    Science.gov (United States)

    Snoeyenbos-West, O.L.; Nevin, K.P.; Anderson, R.T.; Lovely, D.R.

    2000-01-01

    Engineered stimulation of Fe(III) has been proposed as a strategy to enhance the immobilization of radioactive and toxic metals in metal-contaminated subsurface environments. Therefore, laboratory and field studies were conducted to determine which microbial populations would respond to stimulation of Fe(III) reduction in the sediments of sandy aquifers. In laboratory studies, the addition of either various organic electron donors or electron shuttle compounds stimulated Fe(III) reduction and resulted in Geobacter sequences becoming important constituents of the Bacterial 16S rDNA sequences that could be detected with PCR amplification and denaturing gradient gel electrophoresis (DGGE). Quantification of Geobacteraceae sequences with a PCR most-probable-number technique indicated that the extent to which numbers of Geobacter increased was related to the degree of stimulation of Fe(III) reduction. Geothrix species were also enriched in some instances, but were orders of magnitude less numerous than Geobacter species. Shewanella species were not detected, even when organic compounds known to be electron donors for Shewanella species were used to stimulate Fe(III) reduction in the sediments. Geobacter species were also enriched in two field experiments in which Fe(III) reduction was stimulated with the addition of benzoate or aromatic hydrocarbons. The apparent growth of Geobacter species concurrent with increased Fe(III) reduction suggests that Geobacter species were responsible for much of the Fe(III) reduction in all of the stimulation approaches evaluated in three geographically distinct aquifers. Therefore, strategies for subsurface remediation that involve enhancing the activity of indigenous Fe(III)-reducing populations in aquifers should consider the physiological properties of Geobacter species in their treatment design.

  7. Production of gold nanoparticles by electrode-respiring Geobacter sulfurreducens biofilms

    Energy Technology Data Exchange (ETDEWEB)

    Tanzil, Abid H.; Sultana, Sujala T.; Saunders, Steven R.; Dohnalkova, Alice C.; Shi, Liang; Davenport, Emily; Ha, Phuc; Beyenal, Haluk

    2016-12-01

    Current chemical syntheses of nanoparticles (NP) has had limited success due to the relatively high environmental cost caused by the use of harsh chemicals requiring necessary purification and size-selective fractionation. Therefore, biological approaches have received recent attention for their potential to overcome these obstacles as a benign synthetic approach. The intrinsic nature of biomolecules present in microorganisms has intrigued researchers to design bottom-up approaches to biosynthesize metal nanoparticles using microorganisms. Most of the literature work has focused on NP synthesis using planktonic cells while the use of biofilms are limited. The goal of this work was to synthesize gold nanoparticles (AuNPs) using electrode respiring Geobacter sulfurreducens biofilms. We found that most of the AuNPs are generated in the extracellular matrix of Geobacter biofilms with an average particle size of 20 nm. The formation of AuNPs was verified using TEM, FTIR and EDX. We also found that the extracellular substances extracted from electrode respiring G. sulfurreducens biofilms can reduce Au3+ to AuNPs. It appears that reducing sugars were involved in bioreduction and synthesis of AuNPs and amine groups acted as the major biomolecules involved in binding. This is first demonstration of AuNPs formation from the extracellular matrix of electrode respiring biofilms.

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

    KAUST Repository

    Zhu, Xiuping

    2012-08-01

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

  9. Charge transport in films of Geobacter sulfurreducens on graphite electrodes as a function of film thickness

    KAUST Repository

    Jana, Partha Sarathi; Katuri, Krishna; Kavanagh, Paul; Kumar, Amit Ravi Pradeep; Leech, Dó nal

    2014-01-01

    Harnessing, and understanding the mechanisms of growth and activity of, biofilms of electroactive bacteria (EAB) on solid electrodes is of increasing interest, for application to microbial fuel and electrolysis cells. Microbial electrochemical cell technology can be used to generate electricity, or higher value chemicals, from organic waste. The capability of biofilms of electroactive bacteria to transfer electrons to solid anodes is a key feature of this emerging technology, yet the electron transfer mechanism is not fully characterized as yet. Acetate oxidation current generated from biofilms of an EAB, Geobacter sulfurreducens, on graphite electrodes as a function of time does not correlate with film thickness. Values of film thickness, and the number and local concentration of electrically connected redox sites within Geobacter sulfurreducens biofilms as well as a charge transport diffusion co-efficient for the biofilm can be estimated from non-turnover voltammetry. The thicker biofilms, of 50 ± 9 μm, display higher charge transport diffusion co-efficient than that in thinner films, as increased film porosity of these films improves ion transport, required to maintain electro-neutrality upon electrolysis. This journal is © the Partner Organisations 2014.

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

    Energy Technology Data Exchange (ETDEWEB)

    REGUERA, GEMMA [Michigan State University

    2014-01-16

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

  11. Role of U(VI) adsorption in U(VI) Reduction by Geobacter species

    International Nuclear Information System (INIS)

    Lovely, Derrick

    2008-01-01

    Previous work had suggested that Acholeplasma palmae has a higher capacity for uranium sorption than other bacteria studied. Sorption studies were performed with cells in suspension in various solutions containing uranium, and results were used to generate uranium-biosorption isotherms. Results from this study showed that the U(VI) sorption capacity of G. uraniireducens was relatively similar in simple solutions, such as sodium chloride or bicarbonate. However, this ability to sorb uranium significantly decreased in groundwater. This suggested that certain chemicals present in the groundwater were inhibiting the ability of cell components of Geobacter to adsorb uranium. It was hypothesized that uranium removal would also be diminished in the bicarbonate solution. However, this did not seem to be the case, as uranium was as easily removed in the bicarbonate solution as in the sodium chloride solution.

  12. Genome-wide analysis of the RpoN regulon in Geobacter sulfurreducens

    Directory of Open Access Journals (Sweden)

    Núñez Cinthia

    2009-07-01

    Full Text Available Abstract Background The role of the RNA polymerase sigma factor RpoN in regulation of gene expression in Geobacter sulfurreducens was investigated to better understand transcriptional regulatory networks as part of an effort to develop regulatory modules for genome-scale in silico models, which can predict the physiological responses of Geobacter species during groundwater bioremediation or electricity production. Results An rpoN deletion mutant could not be obtained under all conditions tested. In order to investigate the regulon of the G. sulfurreducens RpoN, an RpoN over-expression strain was made in which an extra copy of the rpoN gene was under the control of a taclac promoter. Combining both the microarray transcriptome analysis and the computational prediction revealed that the G. sulfurreducens RpoN controls genes involved in a wide range of cellular functions. Most importantly, RpoN controls the expression of the dcuB gene encoding the fumarate/succinate exchanger, which is essential for cell growth with fumarate as the terminal electron acceptor in G. sulfurreducens. RpoN also controls genes, which encode enzymes for both pathways of ammonia assimilation that is predicted to be essential under all growth conditions in G. sulfurreducens. Other genes that were identified as part of the RpoN regulon using either the computational prediction or the microarray transcriptome analysis included genes involved in flagella biosynthesis, pili biosynthesis and genes involved in central metabolism enzymes and cytochromes involved in extracellular electron transfer to Fe(III, which are known to be important for growth in subsurface environment or electricity production in microbial fuel cells. The consensus sequence for the predicted RpoN-regulated promoter elements is TTGGCACGGTTTTTGCT. Conclusion The G. sulfurreducens RpoN is an essential sigma factor and a global regulator involved in a complex transcriptional network controlling a variety of

  13. Importance of c-Type cytochromes for U(VI reduction by Geobacter sulfurreducens

    Directory of Open Access Journals (Sweden)

    Leang Ching

    2007-03-01

    Full Text Available Abstract Background In order to study the mechanism of U(VI reduction, the effect of deleting c-type cytochrome genes on the capacity of Geobacter sulfurreducens to reduce U(VI with acetate serving as the electron donor was investigated. Results The ability of several c-type cytochrome deficient mutants to reduce U(VI was lower than that of the wild type strain. Elimination of two confirmed outer membrane cytochromes and two putative outer membrane cytochromes significantly decreased (ca. 50–60% the ability of G. sulfurreducens to reduce U(VI. Involvement in U(VI reduction did not appear to be a general property of outer membrane cytochromes, as elimination of two other confirmed outer membrane cytochromes, OmcB and OmcC, had very little impact on U(VI reduction. Among the periplasmic cytochromes, only MacA, proposed to transfer electrons from the inner membrane to the periplasm, appeared to play a significant role in U(VI reduction. A subpopulation of both wild type and U(VI reduction-impaired cells, 24–30%, accumulated amorphous uranium in the periplasm. Comparison of uranium-accumulating cells demonstrated a similar amount of periplasmic uranium accumulation in U(VI reduction-impaired and wild type G. sulfurreducens. Assessment of the ability of the various suspensions to reduce Fe(III revealed no correlation between the impact of cytochrome deletion on U(VI reduction and reduction of Fe(III hydroxide and chelated Fe(III. Conclusion This study indicates that c-type cytochromes are involved in U(VI reduction by Geobacter sulfurreducens. The data provide new evidence for extracellular uranium reduction by G. sulfurreducens but do not rule out the possibility of periplasmic uranium reduction. Occurrence of U(VI reduction at the cell surface is supported by the significant impact of elimination of outer membrane cytochromes on U(VI reduction and the lack of correlation between periplasmic uranium accumulation and the capacity for uranium

  14. Biochemical Mechanisms and Energy Strategies of Geobacter sulfurreducens for Long- Term Survival

    Science.gov (United States)

    Helmus, R. A.; Liermann, L. J.; Brantley, S. L.; Tien, M.

    2008-12-01

    Numerous species of bacteria have been observed to exhibit a growth advantage in stationary phase (GASP) phenotype, indicating that microorganisms starved of an energy source may adapt to allow for long-term survival. Understanding how Geobacter sulfurreducens persists using various metal forms as energy sources and whether a GASP phenotype develops during long-term growth are important for efficient application of this bacterium to sites requiring engineered bioremediation of soluble metals. Thus, we investigated the growth kinetics and survival of G. sulfurreducens. The growth rate of G. sulfurreducens was highest when cultured with soluble iron and generally higher on iron oxide than manganese oxide, suggesting that soluble metal forms are more readily utilized as energy sources by G. sulfurreducens. By monitoring the abundance of G. sulfurreducens in batch cultures for >6 months, distinct growth, stationary, and prolonged starvation phases were observed and a cell density of 105- 106 cells/mL persisted under long-term starvation conditions. The outgrowth of an aged G. sulfurreducens strain co-cultured with a young strain was monitored as a measure of the existence of the GASP phenotype. As the strains aged, the rpoS gene was cloned and sequenced at different stages of growth to identify mutations corresponding to a growth advantage. The results of these studies provide insight into the use of various metal forms for growth by G. sulfurreducens and its ability to persist when starved of energy sources.

  15. Control of nanoparticle size, reactivity and magnetic properties during the bioproduction of magnetite by Geobacter sulfurreducens

    Energy Technology Data Exchange (ETDEWEB)

    Byrne, J. M.; Telling, N. D.; Coker, V. S.; Pattrick, R. A. D.; Laan, G. van der; Arenholz, E.; Tuna, F.; Lloyd, J. R.

    2011-08-02

    The bioproduction of nano-scale magnetite by Fe(III)-reducing bacteria offers a potentially tunable, environmentally benign route to magnetic nanoparticle synthesis. Here, we demonstrate that it is possible to control the size of magnetite nanoparticles produced by Geobacter sulfurreducens, by adjusting the total biomass introduced at the start of the process. The particles have a narrow size distribution and can be controlled within the range of 10-50 nm. X-ray diffraction analysis indicates that controlled production of a number of different biominerals is possible via this method including goethite, magnetite and siderite, but their formation is strongly dependent upon the rate of Fe(III) reduction and total concentration and rate of Fe(II) produced by the bacteria during the reduction process. Relative cation distributions within the structure of the nanoparticles has been investigated by X-ray magnetic circular dichroism and indicates the presence of a highly reduced surface layer which is not observed when magnetite is produced through abiotic methods. The enhanced Fe(II)-rich surface, combined with small particle size, has important environmental applications such as in the reductive bioremediation of organics, radionuclides and metals. In the case of Cr(VI), as a model high-valence toxic metal, optimised biogenic magnetite is able to reduce and sequester the toxic hexavalent chromium very efficiently in the less harmful trivalent form.

  16. Computational and experimental analysis of redundancy in the central metabolism of Geobacter sulfurreducens.

    Directory of Open Access Journals (Sweden)

    Daniel Segura

    2008-02-01

    Full Text Available Previous model-based analysis of the metabolic network of Geobacter sulfurreducens suggested the existence of several redundant pathways. Here, we identified eight sets of redundant pathways that included redundancy for the assimilation of acetate, and for the conversion of pyruvate into acetyl-CoA. These equivalent pathways and two other sub-optimal pathways were studied using 5 single-gene deletion mutants in those pathways for the evaluation of the predictive capacity of the model. The growth phenotypes of these mutants were studied under 12 different conditions of electron donor and acceptor availability. The comparison of the model predictions with the resulting experimental phenotypes indicated that pyruvate ferredoxin oxidoreductase is the only activity able to convert pyruvate into acetyl-CoA. However, the results and the modeling showed that the two acetate activation pathways present are not only active, but needed due to the additional role of the acetyl-CoA transferase in the TCA cycle, probably reflecting the adaptation of these bacteria to acetate utilization. In other cases, the data reconciliation suggested additional capacity constraints that were confirmed with biochemical assays. The results demonstrate the need to experimentally verify the activity of key enzymes when developing in silico models of microbial physiology based on sequence-based reconstruction of metabolic networks.

  17. Visible spectroelectrochemical characterization of Geobacter sulfurreducens biofilms on optically transparent indium tin oxide electrode

    International Nuclear Information System (INIS)

    Jain, Anand; Gazzola, Giulio; Panzera, Aurora; Zanoni, Michele; Marsili, Enrico

    2011-01-01

    We report visible spectroelectrochemical (SEC) characterization of cytochrome c 552 (cyt c 552 ) in viable Geobacter sulfurreducens biofilms on tin-doped indium oxide (ITO) electrodes poised at 0.24 V vs. SHE. G. sulfurreducens biofilms were grown in minimal medium with acetate as electron donor (turnover conditions), followed by 24 h incubation in electron donor-depleted medium (non-turnover conditions). The electronic absorption spectra of G. sulfurreducens biofilms showed the lowest energy absorption band in the reduced state at 552 nm, which indicated excess of cyt c 552 in the biofilm. The spectra under non-turnover conditions displayed gradual reduction of the cyt c 552 , following the step-wise decrease of electrode potential from 0.0 V to −0.6 V vs. standard calomel electrode (SCE). The spectral changes were fully reversible in both positive and negative direction of the scan potential, with average midpoint potential value of −0.42 V vs. SCE. Confocal microscopy analysis revealed that the thickness of biofilms under turnover conditions and non-turnover conditions was approximately 35 and 3.5 μm, respectively. This is the first study to observe the reversible redox conversion of cyt c 552 in viable G. sulfurreducens biofilms.

  18. Investigating different mechanisms for biogenic selenite transformations: Geobacter sulfurreducens, Shewanella oneidensis and Veillonella atypica

    Science.gov (United States)

    Pearce, C.I.; Pattrick, R.A.D.; Law, N.; Charnock, J.M.; Coker, V.S.; Fellowes, J.W.; Oremland, R.S.; Lloyd, J.R.

    2009-01-01

    The metal-reducing bacteria Geobacter sulfurreducens, Shewanella oneidensis and Veillonella atypica, use different mechanisms to transform toxic, bioavailable sodium selenite to less toxic, non-mobile elemental selenium and then to selenide in anaerobic environments, offering the potential for in situ and ex situ bioremediation of contaminated soils, sediments, industrial effluents, and agricultural drainage waters. The products of these reductive transformations depend on both the organism involved and the reduction conditions employed, in terms of electron donor and exogenous extracellular redox mediator. The intermediary phase involves the precipitation of elemental selenium nanospheres and the potential role of proteins in the formation of these structures is discussed. The bionanomineral phases produced during these transformations, including both elemental selenium nanospheres and metal selenide nanoparticles, have catalytic, semiconducting and light-emitting properties, which may have unique applications in the realm of nanophotonics. This research offers the potential to combine remediation of contaminants with the development of environmentally friendly manufacturing pathways for novel bionanominerals. ?? 2009 Taylor & Francis.

  19. Development of a biomarker for Geobacter activity and strain composition: Proteogenomic analysis of the citrate synthase protein during bioremediation of U(VI)

    Energy Technology Data Exchange (ETDEWEB)

    Wilkins, M.J.; Callister, S.J.; Miletto, M.; Williams, K.H.; Nicora, C.D.; Lovley, D.R.; Long, P.E.; Lipton, M.S.

    2010-02-15

    Monitoring the activity of target microorganisms during stimulated bioremediation is a key problem for the development of effective remediation strategies. At the US Department of Energy's Integrated Field Research Challenge (IFRC) site in Rifle, CO, the stimulation of Geobacter growth and activity via subsurface acetate addition leads to precipitation of U(VI) from groundwater as U(IV). Citrate synthase (gltA) is a key enzyme in Geobacter central metabolism that controls flux into the TCA cycle. Here, we utilize shotgun proteomic methods to demonstrate that the measurement of gltA peptides can be used to track Geobacter activity and strain evolution during in situ biostimulation. Abundances of conserved gltA peptides tracked Fe(III) reduction and changes in U(VI) concentrations during biostimulation, whereas changing patterns of unique peptide abundances between samples suggested sample-specific strain shifts within the Geobacter population. Abundances of unique peptides indicated potential differences at the strain level between Fe(III)-reducing populations stimulated during in situ biostimulation experiments conducted a year apart at the Rifle IFRC. These results offer a novel technique for the rapid screening of large numbers of proteomic samples for Geobacter species and will aid monitoring of subsurface bioremediation efforts that rely on metal reduction for desired outcomes.

  20. Electrochemical activities of Geobacter biofilms growing on electrodes with various potentials

    International Nuclear Information System (INIS)

    Li, Dao-Bo; Huang, Yu-Xi; Li, Jie; Li, Ling-Li; Tian, Li-Jiao; Yu, Han-Qing

    2017-01-01

    Highlights: • Dependence of current generation on potentials by G. sulfurreducens is complex with the optimum at +0.1 V. • Unfavorable spatial distribution of biological activity within the biofilm at high potentials limits the current generation. • Same cytochrome c species are used for electron transfer in the biofilms developed at all potentials. - Abstract: Exoelectrogenic bacteria (EEB) play a central role in bioenergy recovery, biogeochemistry of elements, and polluting remediation. The electrochemical activity of EEB biofilm on electrode was proven to be dependent on the electrode potential, but the mechanism behind such a phenomenon is unclear. In this work, Geobacter sulfurreducens biofilms were developed at potentials ranging from −0.1 V to +0.6 V vs. standard hydrogen electrode to explore the profiles of potential regulation on G. sulfurreducens biofilm development and the electrochemical activity. We found that elevating the developing potential could improve the current generation by G. sulfurreducens biofilm until +0.1 V. At higher potentials less current was generated, although more biomass was formed on the electrode. The same cytochrome c species were synthesized for electron transfer in all biofilms, independent of the developing potential. Electrochemical experimental results and redox-sensitive staining imagings proved that the biofilms developed at +0.2 V–+0.4 V had greater cytochrome c contents and reducing capacities than the others. Current generation at high potentials was likely to be limited by both the metabolic rate and the electron transfer kinetics. These findings are useful for tuning the electrochemical activity of biofilm in catalyzing redox processes or generating electricity, which is crucial for the environmental and electrochemical application of EEB.

  1. Geobacter sulfurreducens adapts to low electrode potential for extracellular electron transfer

    International Nuclear Information System (INIS)

    Peng, Luo; Zhang, Xiao-Ting; Yin, Jie; Xu, Shuo-Yuan; Zhang, Yong; Xie, De-Ti; Li, Zhen-Lun

    2016-01-01

    Microbial extracellular electron transfer (EET) occurring in natural and engineering processes is attracting increasing interests. While a meaningful question for bioenergetics, microbial physiology and microbial electrochemical systems; less is known about the lower limit of electron acceptor reduction potential for EET. It is also unclear how microbes adapt to weak electron acceptors. This study evaluated Geobacter sulfurreducens biofilms grown with an electrode poised at −0.25 V vs. SHE. This potential was found to be sufficient for microbial metabolism and proliferation. The turnover cyclic voltammetries found that these biofilms had a half-saturation potential of −0.242 ± 0.004 V, in contrast to −0.151 ± 0.003 V for that of the biofilms grown under 0.2 V. For the biofilms grown under 0.2 V, differential pulse voltammetry showed that the metabolic current was mediated by interfacial cofactors with mid-point potential around −0.16 V performing single-electron electron transfer (ET). The major electron conduits for the biofilms respiring under −0.25 V had mid-point potentials of −0.22 V or −0.26 V, which appeared to perform two-electron ET. Under the non-turnover condition, both biofilms showed similar patterns in voltammograms and the low-potential conduits largely disappeared for the biofilms grown under −0.25 V. Transcriptome analysis identified 17 cytochrome-c genes significantly up-regulated for the biofilms grown under −0.25 V, together with many other genes linked to the ET system. It was also noted that, lowering the poised potential from −0.25 V to −0.28 V (the fuel standard oxidation potential) did not fully inhibit microbial respiration.

  2. MacA is a second cytochrome c peroxidase of Geobacter sulfurreducens.

    Science.gov (United States)

    Seidel, Julian; Hoffmann, Maren; Ellis, Katie E; Seidel, Antonia; Spatzal, Thomas; Gerhardt, Stefan; Elliott, Sean J; Einsle, Oliver

    2012-04-03

    The metal-reducing δ-proteobacterium Geobacter sulfurreducens produces a large number of c-type cytochromes, many of which have been implicated in the transfer of electrons to insoluble metal oxides. Among these, the dihemic MacA was assigned a central role. Here we have produced G. sulfurreducens MacA by recombinant expression in Escherichia coli and have solved its three-dimensional structure in three different oxidation states. Sequence comparisons group MacA into the family of diheme cytochrome c peroxidases, and the protein indeed showed hydrogen peroxide reductase activity with ABTS(-2) as an electron donor. The observed K(M) was 38.5 ± 3.7 μM H(2)O(2) and v(max) was 0.78 ± 0.03 μmol of H(2)O(2)·min(-1)·mg(-1), resulting in a turnover number k(cat) = 0.46 · s(-1). In contrast, no Fe(III) reductase activity was observed. MacA was found to display electrochemical properties similar to other bacterial diheme peroxidases, in addition to the ability to electrochemically mediate electron transfer to the soluble cytochrome PpcA. Differences in activity between CcpA and MacA can be rationalized with structural variations in one of the three loop regions, loop 2, that undergoes conformational changes during reductive activation of the enzyme. This loop is adjacent to the active site heme and forms an open loop structure rather than a more rigid helix as in CcpA. For the activation of the protein, the loop has to displace the distal ligand to the active site heme, H93, in loop 1. A H93G variant showed an unexpected formation of a helix in loop 2 and disorder in loop 1, while a M297H variant that altered the properties of the electron transfer heme abolished reductive activation.

  3. Metabolic response of Geobacter sulfurreducens towards electron donor/acceptor variation

    Directory of Open Access Journals (Sweden)

    Lovley Derek R

    2010-11-01

    Full Text Available Abstract Background Geobacter sulfurreducens is capable of coupling the complete oxidation of organic compounds to iron reduction. The metabolic response of G. sulfurreducens towards variations in electron donors (acetate, hydrogen and acceptors (Fe(III, fumarate was investigated via 13C-based metabolic flux analysis. We examined the 13C-labeling patterns of proteinogenic amino acids obtained from G. sulfurreducens cultured with 13C-acetate. Results Using 13C-based metabolic flux analysis, we observed that donor and acceptor variations gave rise to differences in gluconeogenetic initiation, tricarboxylic acid cycle activity, and amino acid biosynthesis pathways. Culturing G. sulfurreducens cells with Fe(III as the electron acceptor and acetate as the electron donor resulted in pyruvate as the primary carbon source for gluconeogenesis. When fumarate was provided as the electron acceptor and acetate as the electron donor, the flux analysis suggested that fumarate served as both an electron acceptor and, in conjunction with acetate, a carbon source. Growth on fumarate and acetate resulted in the initiation of gluconeogenesis by phosphoenolpyruvate carboxykinase and a slightly elevated flux through the oxidative tricarboxylic acid cycle as compared to growth with Fe(III as the electron acceptor. In addition, the direction of net flux between acetyl-CoA and pyruvate was reversed during growth on fumarate relative to Fe(III, while growth in the presence of Fe(III and acetate which provided hydrogen as an electron donor, resulted in decreased flux through the tricarboxylic acid cycle. Conclusions We gained detailed insight into the metabolism of G. sulfurreducens cells under various electron donor/acceptor conditions using 13C-based metabolic flux analysis. Our results can be used for the development of G. sulfurreducens as a chassis for a variety of applications including bioremediation and renewable biofuel production.

  4. Identification of Genes Involved in Biofilm Formation and Respiration via Mini-Himar Transposon Mutagenesis of Geobacter sulfurreducens▿ †

    Science.gov (United States)

    Rollefson, Janet B.; Levar, Caleb E.; Bond, Daniel R.

    2009-01-01

    Electron transfer from cells to metals and electrodes by the Fe(III)-reducing anaerobe Geobacter sulfurreducens requires proper expression of redox proteins and attachment mechanisms to interface bacteria with surfaces and neighboring cells. We hypothesized that transposon mutagenesis would complement targeted knockout studies in Geobacter spp. and identify novel genes involved in this process. Escherichia coli mating strains and plasmids were used to develop a conjugation protocol and deliver mini-Himar transposons, creating a library of over 8,000 mutants that was anaerobically arrayed and screened for a range of phenotypes, including auxotrophy for amino acids, inability to reduce Fe(III) citrate, and attachment to surfaces. Following protocol validation, mutants with strong phenotypes were further characterized in a three-electrode system to simultaneously quantify attachment, biofilm development, and respiratory parameters, revealing mutants defective in Fe(III) reduction but unaffected in electron transfer to electrodes (such as an insertion in GSU1330, a putative metal export protein) or defective in electrode reduction but demonstrating wild-type biofilm formation (due to an insertion upstream of the NHL domain protein GSU2505). An insertion in a putative ATP-dependent transporter (GSU1501) eliminated electrode colonization but not Fe(III) citrate reduction. A more complex phenotype was demonstrated by a mutant containing an insertion in a transglutaminase domain protein (GSU3361), which suddenly ceased to respire when biofilms reached approximately 50% of the wild-type levels. As most insertions were not in cytochromes but rather in transporters, two-component signaling proteins, and proteins of unknown function, this collection illustrates how biofilm formation and electron transfer are separate but complementary phenotypes, controlled by multiple loci not commonly studied in Geobacter spp. PMID:19395486

  5. Geobacter Dominates the Inner Layers of a Stratified Biofilm on a Fluidized Anode During Brewery Wastewater Treatment

    Directory of Open Access Journals (Sweden)

    Sara Tejedor-Sanz

    2018-03-01

    Full Text Available In this study, we designed a microbial electrochemical fluidized bed reactor (ME-FBR, with an electroconductive anodic bed made of activated carbon particles for treating a brewery wastewater. Under a batch operating mode, acetate and propionate consumption rates were 13-fold and 2.4-fold higher, respectively, when the fluidized anode was polarized (0.2 V with respect to open circuit conditions. Operating in a continuous mode, this system could effectively treat the brewery effluent at organic loading rates (OLR over 1.7 kg m-3NRV d-1 and with removal efficiencies of 95 ± 1.4% (hydraulic retention time of 1 day and an influent of 1.7 g-COD L-1. The coulombic efficiency values highly depended upon the OLR applied, and varied from a 56 ± 15% to 10 ± 1%. Fluorescence in situ hybridization (FISH analysis revealed a relative high abundance of Geobacter species (ca. 20%, and clearly showed a natural microbial stratification. Interestingly, the Geobacter cluster was highly enriched in the innermost layers of the biofilm (thickness of 10 μm, which were in contact with the electroconductive particles of bed, whereas the rest of bacteria were located in the outermost layers. To our knowledge, this is the first time that such a clear microbial stratification has been observed on an anode-respiring biofilm. Our results revealed the relevant role of Geobacter in switching between the electrode and other microbial communities performing metabolic reactions in the outermost environment of the biofilm.

  6. MOLECULAR DOCKING AND DYNAMICS STUDIES ON THE PROTEIN-PROTEIN INTERACTIONS OF ELECTRICALLY ACTIVE PILIN NANOWIRES OF GEOBACTER SULFURREDUCENS.

    Directory of Open Access Journals (Sweden)

    D. Jeya Sundara Sharmila1 *

    2017-06-01

    Full Text Available Molecular interactions are key aspects in biological recognitions applicable in nano/micro systems. Bacterial nanowires are pilus filament based structures that can conduct electrons. The transport of electron is proposed to be facilitated by filamentous fibers made up of polymeric assemblies of proteins called pilin. Geobacter sulfurreducens is capable of delivering electrons through extracellular electron transport (EET by employing conductive nanowires, which are pilin proteins composed of type IV subunit PilA. Protein-protein interactions play an important role in the stabilization of the pilin nanowire assembly complex and it contains transmembrane (TM domain. In current study, protein-protein docking and multiple molecular dynamic (MD simulations were performed to understand the binding mode of pilin nanowires. The MD result explains the conformational behavior and folding of pilin nanowires in water environment in different time scale duration 20, 5, 5, 10 and 20ns (total of 60ns. Direct hydrogen bonds and water mediated hydrogen bonds that play a crucial role during the simulation were investigated. The conformational state, folding, end-toend distance profile and hydrogen bonding behavior had indicated that the Geobacter sulfurreducens pilin nanowires have electrical conductivity properties.

  7. Functional Role of Infective Viral Particles on Metal Reduction

    Energy Technology Data Exchange (ETDEWEB)

    Coates, John D.

    2014-04-01

    A proposed strategy for the remediation of uranium (U) contaminated sites was based on the immobilization of U by reducing the oxidized soluble U, U(VI), to form a reduced insoluble end product, U(IV). Previous studies identified Geobacter sp., including G. sulfurreducens and G. metallireducens, as predominant U(VI)-reducing bacteria under acetate-oxidizing and U(VI)-reducing conditions. Examination of the finished genome sequence annotation of the canonical metal reducing species Geobacter sulfurreducens strain PCA and G. metallireduceans strain GS-15 as well as the draft genome sequence of G. uraniumreducens strain Rf4 identified phage related proteins. In addition, the completed genome for Anaeromyxobacter dehalogenans and the draft genome sequence of Desulfovibrio desulfuricans strain G20, two more model metal-reducing bacteria, also revealed phage related sequences. The presence of these gene sequences indicated that Geobacter spp., Anaeromyxobacter spp., and Desulfovibrio spp. are susceptible to viral infection. Furthermore, viral populations in soils and sedimentary environments in the order of 6.4×10{sup 6}–2.7×10{sup 10} VLP’s cm{sup -3} have been observed. In some cases, viral populations exceed bacterial populations in these environments suggesting that a relationship may exist between viruses and bacteria. Our preliminary screens of samples collected from the ESR FRC indicated that viral like particles were observed in significant numbers. The objective of this study was to investigate the potential functional role viruses play in metal reduction specifically Fe(III) and U(VI) reduction, the environmental parameters affecting viral infection of metal reducing bacteria, and the subsequent effects on U transport.

  8. Anode biofilm transcriptomics reveals outer surface components essential for high density current production in Geobacter sulfurreducens fuel cells.

    Directory of Open Access Journals (Sweden)

    Kelly P Nevin

    Full Text Available The mechanisms by which Geobacter sulfurreducens transfers electrons through relatively thick (>50 microm biofilms to electrodes acting as a sole electron acceptor were investigated. Biofilms of Geobacter sulfurreducens were grown either in flow-through systems with graphite anodes as the electron acceptor or on the same graphite surface, but with fumarate as the sole electron acceptor. Fumarate-grown biofilms were not immediately capable of significant current production, suggesting substantial physiological differences from current-producing biofilms. Microarray analysis revealed 13 genes in current-harvesting biofilms that had significantly higher transcript levels. The greatest increases were for pilA, the gene immediately downstream of pilA, and the genes for two outer c-type membrane cytochromes, OmcB and OmcZ. Down-regulated genes included the genes for the outer-membrane c-type cytochromes, OmcS and OmcT. Results of quantitative RT-PCR of gene transcript levels during biofilm growth were consistent with microarray results. OmcZ and the outer-surface c-type cytochrome, OmcE, were more abundant and OmcS was less abundant in current-harvesting cells. Strains in which pilA, the gene immediately downstream from pilA, omcB, omcS, omcE, or omcZ was deleted demonstrated that only deletion of pilA or omcZ severely inhibited current production and biofilm formation in current-harvesting mode. In contrast, these gene deletions had no impact on biofilm formation on graphite surfaces when fumarate served as the electron acceptor. These results suggest that biofilms grown harvesting current are specifically poised for electron transfer to electrodes and that, in addition to pili, OmcZ is a key component in electron transfer through differentiated G. sulfurreducens biofilms to electrodes.

  9. Geobacter daltonii sp. nov., an Fe(III)- and uranium(VI)-reducing bacterium isolated from a shallow subsurface exposed to mixed heavy metal and hydrocarbon contamination.

    Science.gov (United States)

    Prakash, Om; Gihring, Thomas M; Dalton, Dava D; Chin, Kuk-Jeong; Green, Stefan J; Akob, Denise M; Wanger, Greg; Kostka, Joel E

    2010-03-01

    An Fe(III)- and uranium(VI)-reducing bacterium, designated strain FRC-32(T), was isolated from a contaminated subsurface of the USA Department of Energy Oak Ridge Field Research Center (ORFRC) in Oak Ridge, Tennessee, where the sediments are exposed to mixed waste contamination of radionuclides and hydrocarbons. Analyses of both 16S rRNA gene and the Geobacteraceae-specific citrate synthase (gltA) mRNA gene sequences retrieved from ORFRC sediments indicated that this strain was abundant and active in ORFRC subsurface sediments undergoing uranium(VI) bioremediation. The organism belonged to the subsurface clade of the genus Geobacter and shared 92-98 % 16S rRNA gene and 75-81 % rpoB gene sequence similarities with other recognized species of the genus. In comparison to its closest relative, Geobacter uraniireducens Rf4(T), according to 16S rRNA gene sequence similarity, strain FRC-32(T) showed a DNA-DNA relatedness value of 21 %. Cells of strain FRC-32(T) were Gram-negative, non-spore-forming, curved rods, 1.0-1.5 microm long and 0.3-0.5 microm in diameter; the cells formed pink colonies in a semisolid cultivation medium, a characteristic feature of the genus Geobacter. The isolate was an obligate anaerobe, had temperature and pH optima for growth at 30 degrees C and pH 6.7-7.3, respectively, and could tolerate up to 0.7 % NaCl although growth was better in the absence of NaCl. Similar to other members of the Geobacter group, strain FRC-32(T) conserved energy for growth from the respiration of Fe(III)-oxyhydroxide coupled with the oxidation of acetate. Strain FRC-32(T) was metabolically versatile and, unlike its closest relative, G. uraniireducens, was capable of utilizing formate, butyrate and butanol as electron donors and soluble ferric iron (as ferric citrate) and elemental sulfur as electron acceptors. Growth on aromatic compounds including benzoate and toluene was predicted from preliminary genomic analyses and was confirmed through successive transfer with

  10. The NASA competitive placement plan for positions GS-15 and below (including trades and labor positions)

    Science.gov (United States)

    1993-01-01

    This plan provides the framework for selection based on merit from among the best qualified candidates available. Selections will be made without regard to political, religious, or labor organization affiliation or nonaffiliation, marital status, race, color, sex, national origin, nondisqualifying disability, or age. This plan does not guarantee promotion but rather ensures that all qualified available candidates receive fair and equitable consideration for positions filled under these competitive procedures. Announcing a vacancy under this plan is only one method of locating applicants for a position and can be used in conjunction with other methods. Subject to applicable law and regulation, selection of an individual to fill a position is the decision of management, as is the decision as to the method(s) to be used in identifying candidates. This plan is applicable to all NASA Installations. It covers all positions in the competitive service at (and below) the GS/GM-15 level (including all trades and labor positions), except positions in the Office of the Inspector General. The requirements herein are not intended to, nor should they be construed to limit in any way, the independent personnel authority of the Inspector General under the Inspector General Act, as Amended.

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

    Science.gov (United States)

    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.

  12. A defined co-culture of Geobacter sulfurreducens and Escherichia coli in a membrane-less microbial fuel cell.

    Science.gov (United States)

    Bourdakos, Nicholas; Marsili, Enrico; Mahadevan, Radhakrishnan

    2014-04-01

    Wastewater-fed microbial fuel cells (MFCs) are a promising technology to treat low-organic carbon wastewater and recover part of the chemical energy in wastewater as electrical power. However, the interactions between electrochemically active and fermentative microorganisms cannot be easily studied in wastewater-fed MFCs because of their complex microbial communities. Defined co-culture MFCs provide a detailed understanding of such interactions. In this study, we characterize the extracellular metabolites in laboratory-scale membrane-less MFCs inoculated with Geobacter sulfurreducens and Escherichia coli co-culture and compare them with pure culture MFCs. G. sulfurreducens MFCs are sparged to maintain anaerobic conditions, while co-culture MFCs rely on E. coli for oxygen removal. G. sulfurreducens MFCs have a power output of 128 mW m(-2) , compared to 63 mW m(-2) from the co-culture MFCs. Analysis of metabolites shows that succinate production in co-culture MFCs decreases current production by G. sulfurreducens and that the removal of succinate is responsible for the increased current density in the late co-culture MFCs. Interestingly, pH adjustment is not required for co-culture MFCs but a base addition is necessary for E. coli MFCs and cultures in vials. Our results show that defined co-culture MFCs provide clear insights into metabolic interactions among bacteria while maintaining a low operational complexity. © 2013 Wiley Periodicals, Inc.

  13. Reduction of low potential electron acceptors requires the CbcL inner membrane cytochrome of Geobacter sulfurreducens.

    Science.gov (United States)

    Zacharoff, Lori; Chan, Chi Ho; Bond, Daniel R

    2016-02-01

    The respiration of metals by the bacterium Geobacter sulfurreducens requires electrons generated by metabolism to pass from the interior of the cell to electron acceptors beyond the cell membranes. The G. sulfurreducens inner membrane multiheme c-type cytochrome ImcH is required for respiration to extracellular electron acceptors with redox potentials greater than -0.1 V vs. SHE, but ImcH is not essential for electron transfer to lower potential acceptors. In contrast, deletion of cbcL, encoding an inner membrane protein consisting of b-type and multiheme c-type cytochrome domains, severely affected reduction of low potential electron acceptors such as Fe(III)-oxides and electrodes poised at -0.1 V vs. SHE. Catalytic cyclic voltammetry of a ΔcbcL strain growing on poised electrodes revealed a 50 mV positive shift in driving force required for electron transfer out of the cell. In non-catalytic conditions, low-potential peaks present in wild type biofilms were absent in ∆cbcL mutants. Expression of cbcL in trans increased growth at low redox potential and restored features to cyclic voltammetry. This evidence supports a model where CbcL is a component of a second electron transfer pathway out of the G. sulfurreducens inner membrane that dominates when redox potential is at or below -0.1 V vs. SHE. Copyright © 2015. Published by Elsevier B.V.

  14. Optimization of culture conditions and electricity generation using Geobacter sulfurreducens in a dual-chambered microbial fuel-cell

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Mi-Sun; Lee, Yu-jin [Bioenergy Research Center, Korea Institute of Energy Research, Daejeon 305-343 (Korea, Republic of)

    2010-12-15

    The promise of generating electricity from the oxidation of organic substances using metal-reducing bacteria is drawing attention as an alternate form of bio-technology with positive environmental implications. In this study, we examined various experimental factors to obtain the maximum power output in a dual-chamber mediator-less microbial fuel-cell (MFC) using Geobacter sulfurreducens and acetate as an electron donor in a semi-continuous mode. The G. sulfurreducens culture conditions were optimized in a nutrient buffer containing 20 mM of acetate and 50 mM of fumarate at pH 6.8 and 30 C. For use in the MFC system, electrodes were made with carbon paper (area: 11.5 cm{sup 2}) and spaced 1.5 cm apart. Once the MFC was inoculated with the pre-cultured G. sulfurreducens in the anode chamber and while air was continuously sparged to the cathode chamber, the cells produced electricity stably over 60 days with the regular addition of 20 mM acetate, generating the maximum power density of 7 mW/m{sup 2} with a 5000 and ohm; load. The current output was significantly increased, by 1.6 times after 20 days of incubation under the same experimental conditions, when the carbon-paper anode was coated with carbon nanotubes. (author)

  15. Contrasting Effects of Dissolved Organic Matter on Mercury Methylation by Geobacter sulfurreducens PCA and Desulfovibrio desulfuricans ND132.

    Science.gov (United States)

    Zhao, Linduo; Chen, Hongmei; Lu, Xia; Lin, Hui; Christensen, Geoff A; Pierce, Eric M; Gu, Baohua

    2017-09-19

    Natural dissolved organic matter (DOM) affects mercury (Hg) redox reactions and anaerobic microbial methylation in the environment. Several studies have shown that DOM can enhance Hg methylation, especially under sulfidic conditions, whereas others show that DOM inhibits Hg methylation due to strong Hg-DOM complexation. In this study, we investigated and compared the effects of DOM on Hg methylation by an iron-reducing bacterium Geobacter sulfurreducens PCA and a sulfate-reducing bacterium Desulfovibrio desulfuricans ND132 under nonsulfidic conditions. The methylation experiment was performed with washed cells either in the absence or presence of DOM or glutathione, both of which form strong complexes with Hg via thiol-functional groups. DOM was found to greatly inhibit Hg methylation by G. Sulfurreducens PCA but enhance Hg methylation by D. desulfuricans ND132 cells with increasing DOM concentration. These strain-dependent opposing effects of DOM were also observed with glutathione, suggesting that thiols in DOM likely played an essential role in affecting microbial Hg uptake and methylation. Additionally, DOM and glutathione greatly decreased Hg sorption by G. sulfurreducens PCA but showed little effect on D. desulfuricans ND132 cells, demonstrating that ND132 has a higher affinity to sorb or take up Hg than the PCA strain. These observations indicate that DOM effects on Hg methylation are bacterial strain specific, depend on the DOM:Hg ratio or site-specific conditions, and may thus offer new insights into the role of DOM in methylmercury production in the environment.

  16. High Resolution AFM and Single-Cell Resonance Raman Spectroscopy of Geobacter sulfurreducens Biofilms Early in Growth

    Energy Technology Data Exchange (ETDEWEB)

    Lebedev, Nikolai, E-mail: nikolai.lebedev@nrl.navy.mil; Strycharz-Glaven, Sarah M.; Tender, Leonard M., E-mail: nikolai.lebedev@nrl.navy.mil [Center for Biomolecular Science and Engineering, US Naval Research Laboratory, Washington, DC (United States)

    2014-08-21

    Atomic force microscopy and confocal resonance Raman microscopy (CRRM) of single-cells were used to study the transition of anode-grown Geobacter sulfurreducens biofilms from lag phase (initial period of low current) to exponential phase (subsequent period of rapidly increasing current). Results reveal that lag phase biofilms consist of lone cells and tightly packed single-cell thick clusters crisscrossed with extracellular linear structures that appears to be comprised of nodules approximately 20 nm in diameter aligned end to end. By early exponential phase, cell clusters expand laterally and a second layer of closely packed cells begins to form on top of the first. Abundance of c-type cytochromes (c-Cyt) is threefold greater in two-cell thick regions than in one-cell thick regions. The results indicate that early biofilm growth involves two transformations. The first is from lone cells to two-dimensionally associated cells during lag phase when current remains low. This is accompanied by formation of extracellular linear structures. The second is from two- to three-dimensionally associated cells during early exponential phase when current begins to increase rapidly. This is accompanied by a dramatic increase in c-Cyt abundance.

  17. High Resolution AFM and Single-Cell Resonance Raman Spectroscopy of Geobacter sulfurreducens Biofilms Early in Growth

    International Nuclear Information System (INIS)

    Lebedev, Nikolai; Strycharz-Glaven, Sarah M.; Tender, Leonard M.

    2014-01-01

    Atomic force microscopy and confocal resonance Raman microscopy (CRRM) of single-cells were used to study the transition of anode-grown Geobacter sulfurreducens biofilms from lag phase (initial period of low current) to exponential phase (subsequent period of rapidly increasing current). Results reveal that lag phase biofilms consist of lone cells and tightly packed single-cell thick clusters crisscrossed with extracellular linear structures that appears to be comprised of nodules approximately 20 nm in diameter aligned end to end. By early exponential phase, cell clusters expand laterally and a second layer of closely packed cells begins to form on top of the first. Abundance of c-type cytochromes (c-Cyt) is threefold greater in two-cell thick regions than in one-cell thick regions. The results indicate that early biofilm growth involves two transformations. The first is from lone cells to two-dimensionally associated cells during lag phase when current remains low. This is accompanied by formation of extracellular linear structures. The second is from two- to three-dimensionally associated cells during early exponential phase when current begins to increase rapidly. This is accompanied by a dramatic increase in c-Cyt abundance.

  18. Fate of Cd during microbial Fe(III) mineral reduction by a novel and Cd-tolerant Geobacter species.

    Science.gov (United States)

    Muehe, E Marie; Obst, Martin; Hitchcock, Adam; Tyliszczak, Tolek; Behrens, Sebastian; Schröder, Christian; Byrne, James M; Michel, F Marc; Krämer, Ute; Kappler, Andreas

    2013-12-17

    Fe(III) (oxyhydr)oxides affect the mobility of contaminants in the environment by providing reactive surfaces for sorption. This includes the toxic metal cadmium (Cd), which prevails in agricultural soils and is taken up by crops. Fe(III)-reducing bacteria can mobilize such contaminants by Fe(III) mineral dissolution or immobilize them by sorption to or coprecipitation with secondary Fe minerals. To date, not much is known about the fate of Fe(III) mineral-associated Cd during microbial Fe(III) reduction. Here, we describe the isolation of a new Geobacter sp. strain Cd1 from a Cd-contaminated field site, where the strain accounts for 10(4) cells g(-1) dry soil. Strain Cd1 reduces the poorly crystalline Fe(III) oxyhydroxide ferrihydrite in the presence of at least up to 112 mg Cd L(-1). During initial microbial reduction of Cd-loaded ferrihydrite, sorbed Cd was mobilized. However, during continuous microbial Fe(III) reduction, Cd was immobilized by sorption to and/or coprecipitation within newly formed secondary minerals that contained Ca, Fe, and carbonate, implying the formation of an otavite-siderite-calcite (CdCO3-FeCO3-CaCO3) mixed mineral phase. Our data shows that microbially mediated turnover of Fe minerals affects the mobility of Cd in soils, potentially altering the dynamics of Cd uptake into food or phyto-remediating plants.

  19. Identification of the electron transfer flavoprotein as an upregulated enzyme in the benzoate utilization of Desulfotignum balticum.

    Science.gov (United States)

    Habe, Hiroshi; Kobuna, Akinori; Hosoda, Akifumi; Kosaka, Tomoyuki; Endoh, Takayuki; Tamura, Hiroto; Yamane, Hisakazu; Nojiri, Hideaki; Omori, Toshio; Watanabe, Kazuya

    2009-07-01

    Desulfotignum balticum utilizes benzoate coupled to sulfate reduction. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis was conducted to detect proteins that increased more after growth on benzoate than on butyrate. A comparison of proteins on 2D gels showed that at least six proteins were expressed. The N-terminal sequences of three proteins exhibited significant identities with the alpha and beta subunits of electron transfer flavoprotein (ETF) from anaerobic aromatic-degraders. By sequence analysis of the fosmid clone insert (37,590 bp) containing the genes encoding the ETF subunits, we identified three genes, whose deduced amino acid sequences showed 58%, 74%, and 62% identity with those of Gmet_2267 (Fe-S oxidoreductase), Gmet_2266 (ETF beta subunit), and Gmet_2265 (ETF alpha subunit) respectively, which exist within the 300-kb genomic island of aromatic-degradation genes from Geobacter metallireducens GS-15. The genes encoding ETF subunits found in this study were upregulated in benzoate utilization.

  20. Electron Transfer Strategies Regulate Carbonate Mineral and Micropore Formation.

    Science.gov (United States)

    Zeng, Zhirui; Tice, Michael M

    2018-01-01

    Some microbial carbonates are robust biosignatures due to their distinct morphologies and compositions. However, whether carbonates induced by microbial iron reduction have such features is unknown. Iron-reducing bacteria use various strategies to transfer electrons to iron oxide minerals (e.g., membrane-bound enzymes, soluble electron shuttles, nanowires, as well as different mechanisms for moving over or attaching to mineral surfaces). This diversity has the potential to create mineral biosignatures through manipulating the microenvironments in which carbonate precipitation occurs. We used Shewanella oneidensis MR-1, Geothrix fermentans, and Geobacter metallireducens GS-15, representing three different strategies, to reduce solid ferric hydroxide in order to evaluate their influence on carbonate and micropore formation (micro-size porosity in mineral rocks). Our results indicate that electron transfer strategies determined the morphology (rhombohedral, spherical, or long-chained) of precipitated calcium-rich siderite by controlling the level of carbonate saturation and the location of carbonate formation. Remarkably, electron transfer strategies also produced distinctive cell-shaped micropores in both carbonate and hydroxide minerals, thus producing suites of features that could potentially serve as biosignatures recording information about the sizes, shapes, and physiologies of iron-reducing organisms. Key Words: Microbial iron reduction-Micropore-Electron transfer strategies-Microbial carbonate. Astrobiology 18, 28-36.

  1. Electron Transfer Strategies Regulate Carbonate Mineral and Micropore Formation

    Science.gov (United States)

    Zeng, Zhirui; Tice, Michael M.

    2018-01-01

    Some microbial carbonates are robust biosignatures due to their distinct morphologies and compositions. However, whether carbonates induced by microbial iron reduction have such features is unknown. Iron-reducing bacteria use various strategies to transfer electrons to iron oxide minerals (e.g., membrane-bound enzymes, soluble electron shuttles, nanowires, as well as different mechanisms for moving over or attaching to mineral surfaces). This diversity has the potential to create mineral biosignatures through manipulating the microenvironments in which carbonate precipitation occurs. We used Shewanella oneidensis MR-1, Geothrix fermentans, and Geobacter metallireducens GS-15, representing three different strategies, to reduce solid ferric hydroxide in order to evaluate their influence on carbonate and micropore formation (micro-size porosity in mineral rocks). Our results indicate that electron transfer strategies determined the morphology (rhombohedral, spherical, or long-chained) of precipitated calcium-rich siderite by controlling the level of carbonate saturation and the location of carbonate formation. Remarkably, electron transfer strategies also produced distinctive cell-shaped micropores in both carbonate and hydroxide minerals, thus producing suites of features that could potentially serve as biosignatures recording information about the sizes, shapes, and physiologies of iron-reducing organisms.

  2. Quantifying Temporal Autocorrelations for the Expression of Geobacter species mRNA Gene Transcripts at Variable Ammonium Levels during in situ U(VI) Bioremediation

    Science.gov (United States)

    Mouser, P. J.

    2010-12-01

    In order to develop decision-making tools for the prediction and optimization of subsurface bioremediation strategies, we must be able to link the molecular-scale activity of microorganisms involved in remediation processes with biogeochemical processes observed at the field-scale. This requires the ability to quantify changes in the in situ metabolic condition of dominant microbes and associate these changes to fluctuations in nutrient levels throughout the bioremediation process. It also necessitates a need to understand the spatiotemporal variability of the molecular-scale information to develop meaningful parameters and constraint ranges in complex bio-physio-chemical models. The expression of three Geobacter species genes (ammonium transporter (amtB), nitrogen fixation (nifD), and a housekeeping gene (recA)) were tracked at two monitoring locations that differed significantly in ammonium (NH4+) concentrations during a field-scale experiment where acetate was injected into the subsurface to simulate Geobacteraceae in a uranium-contaminated aquifer. Analysis of amtB and nifD mRNA transcript levels indicated that NH4+ was the primary form of fixed nitrogen during bioremediation. Overall expression levels of amtB were on average 8-fold higher at NH4+ concentrations of 300 μM or more than at lower NH4+ levels (average 60 μM). The degree of temporal correlation in Geobacter species mRNA expression levels was calculated at both locations using autocorrelation methods that describe the relationship between sample semi-variance and time lag. At the monitoring location with lower NH4+, a temporal correlation lag of 8 days was observed for both amtB and nifD transcript patterns. At the location where higher NH4+ levels were observed, no discernable temporal correlation lag above the sampling frequency (approximately every 2 days) was observed for amtB or nifD transcript fluctuations. Autocorrelation trends in recA expression levels at both locations indicated that

  3. A novel Geobacteraceae-specific outer membrane protein J (OmpJ is essential for electron transport to Fe (III and Mn (IV oxides in Geobacter sulfurreducens

    Directory of Open Access Journals (Sweden)

    Schiffer Marianne

    2005-07-01

    Full Text Available Abstract Background Metal reduction is thought to take place at or near the bacterial outer membrane and, thus, outer membrane proteins in the model dissimilatory metal-reducing organism Geobacter sulfurreducens are of interest to understand the mechanisms of Fe(III reduction in the Geobacter species that are the predominant Fe(III reducers in many environments. Previous studies have implicated periplasmic and outer membrane cytochromes in electron transfer to metals. Here we show that the most abundant outer membrane protein of G. sulfurreducens, OmpJ, is not a cytochrome yet it is required for metal respiration. Results When outer membrane proteins of G. sulfurreducens were separated via SDS-PAGE, one protein, designated OmpJ (outer membrane protein J, was particularly abundant. The encoding gene, which was identified from mass spectrometry analysis of peptide fragments, is present in other Geobacteraceae, but not in organisms outside this family. The predicted localization and structure of the OmpJ protein suggested that it was a porin. Deletion of the ompJ gene in G. sulfurreducens produced a strain that grew as well as the wild-type strain with fumarate as the electron acceptor but could not grow with metals, such as soluble or insoluble Fe (III and insoluble Mn (IV oxide, as the electron acceptor. The heme c content in the mutant strain was ca. 50% of the wild-type and there was a widespread loss of multiple cytochromes from soluble and membrane fractions. Transmission electron microscopy analyses of mutant cells revealed an unusually enlarged periplasm, which is likely to trigger extracytoplasmic stress response mechanisms leading to the degradation of periplasmic and/or outer membrane proteins, such as cytochromes, required for metal reduction. Thus, the loss of the capacity for extracellular electron transport in the mutant could be due to the missing c-type cytochromes, or some more direct, but as yet unknown, role of OmpJ in metal

  4. Changes in phosphorylation of adenosine phosphate and redox state of nicotinamide-adenine dinucleotide (phosphate) in Geobacter sulfurreducens in response to electron acceptor and anode potential variation

    KAUST Repository

    Rose, Nicholas D.; Regan, John M.

    2015-01-01

    © 2015 Elsevier B.V. Geobacter sulfurreducens is one of the dominant bacterial species found in biofilms growing on anodes in bioelectrochemical systems. The intracellular concentrations of reduced and oxidized forms of nicotinamide-adenine dinucleotide (NADH and NAD+, respectively) and nicotinamide-adenine dinucleotide phosphate (NADPH and NADP+, respectively) as well as adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) were measured in G. sulfurreducens using fumarate, Fe(III)-citrate, or anodes poised at different potentials (110, 10, -90, and -190mV (vs. SHE)) as the electron acceptor. The ratios of CNADH/CNAD+ (0.088±0.022) and CNADPH/CNADP+ (0.268±0.098) were similar under all anode potentials tested and with Fe(III)-citrate (reduced extracellularly). Both ratios significantly increased with fumarate as the electron acceptor (0.331±0.094 for NAD and 1.96±0.37 for NADP). The adenylate energy charge (the fraction of phosphorylation in intracellular adenosine phosphates) was maintained near 0.47 under almost all conditions. Anode-growing biofilms demonstrated a significantly higher molar ratio of ATP/ADP relative to suspended cultures grown on fumarate or Fe(III)-citrate. These results provide evidence that the cellular location of reduction and not the redox potential of the electron acceptor controls the intracellular redox potential in G. sulfurreducens and that biofilm growth alters adenylate phosphorylation.

  5. Abundance of the multiheme c-type cytochrome OmcB increases in outer biofilm layers of electrode-grown Geobacter sulfurreducens.

    Directory of Open Access Journals (Sweden)

    Camille S Stephen

    Full Text Available 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.

  6. Changes in phosphorylation of adenosine phosphate and redox state of nicotinamide-adenine dinucleotide (phosphate) in Geobacter sulfurreducens in response to electron acceptor and anode potential variation

    KAUST Repository

    Rose, Nicholas D.

    2015-12-01

    © 2015 Elsevier B.V. Geobacter sulfurreducens is one of the dominant bacterial species found in biofilms growing on anodes in bioelectrochemical systems. The intracellular concentrations of reduced and oxidized forms of nicotinamide-adenine dinucleotide (NADH and NAD+, respectively) and nicotinamide-adenine dinucleotide phosphate (NADPH and NADP+, respectively) as well as adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) were measured in G. sulfurreducens using fumarate, Fe(III)-citrate, or anodes poised at different potentials (110, 10, -90, and -190mV (vs. SHE)) as the electron acceptor. The ratios of CNADH/CNAD+ (0.088±0.022) and CNADPH/CNADP+ (0.268±0.098) were similar under all anode potentials tested and with Fe(III)-citrate (reduced extracellularly). Both ratios significantly increased with fumarate as the electron acceptor (0.331±0.094 for NAD and 1.96±0.37 for NADP). The adenylate energy charge (the fraction of phosphorylation in intracellular adenosine phosphates) was maintained near 0.47 under almost all conditions. Anode-growing biofilms demonstrated a significantly higher molar ratio of ATP/ADP relative to suspended cultures grown on fumarate or Fe(III)-citrate. These results provide evidence that the cellular location of reduction and not the redox potential of the electron acceptor controls the intracellular redox potential in G. sulfurreducens and that biofilm growth alters adenylate phosphorylation.

  7. Global Proteome Response to Deletion of Genes Related to Mercury Methylation and Dissimilatory Metal Reduction Reveals Changes in Respiratory Metabolism in Geobacter sulfurreducens PCA.

    Science.gov (United States)

    Qian, Chen; Johs, Alexander; Chen, Hongmei; Mann, Benjamin F; Lu, Xia; Abraham, Paul E; Hettich, Robert L; Gu, Baohua

    2016-10-07

    Geobacter sulfurreducens PCA can reduce, sorb, and methylate mercury (Hg); however, the underlying biochemical mechanisms of these processes and interdependent metabolic pathways remain unknown. In this study, shotgun proteomics was used to compare global proteome profiles between wild-type G. sulfurreducens PCA and two mutant strains: a ΔhgcAB mutant, which is deficient in two genes known to be essential for Hg methylation and a ΔomcBESTZ mutant, which is deficient in five outer membrane c-type cytochromes and thus impaired in its ability for dissimilatory metal ion reduction. We were able to delineate the global response of G. sulfurreducens PCA in both mutants and identify cellular networks and metabolic pathways that were affected by the loss of these genes. Deletion of hgcAB increased the relative abundances of proteins implicated in extracellular electron transfer, including most of the c-type cytochromes, PilA-C, and OmpB, and is consistent with a previously observed increase in Hg reduction in the ΔhgcAB mutant. Deletion of omcBESTZ was found to significantly increase relative abundances of various methyltransferases, suggesting that a loss of dissimilatory reduction capacity results in elevated activity among one-carbon (C1) metabolic pathways and thus increased methylation. We show that G. sulfurreducens PCA encodes only the folate branch of the acetyl-CoA pathway, and proteins associated with the folate branch were found at lower abundance in the ΔhgcAB mutant strain than the wild type. This observation supports the hypothesis that the function of HgcA and HgcB is linked to C1 metabolism through the folate branch of the acetyl-CoA pathway by providing methyl groups required for Hg methylation.

  8. Direct Involvement of ombB, omaB and omcB Genes in Extracellular Reduction of Fe(III by Geobacter sulfurreducens PCA

    Directory of Open Access Journals (Sweden)

    Yimo eLiu

    2015-10-01

    Full Text Available The tandem gene clusters orfR-ombB-omaB-omcB and orfS-ombC-omaC-omcC of the metal-reducing bacterium Geobacter sulfurreducens PCA are responsible for trans-outer membrane electron transfer during extracellular reduction of Fe(III-citrate and ferrihydrite [a poorly crystalline Fe(III oxide]. Each gene cluster encodes a putative transcriptional factor (OrfR/OrfS, a porin-like outer-membrane protein (OmbB/OmbC, a periplasmic c-type cytochrome (c-Cyt, OmaB/OmaC and an outer-membrane c-Cyt (OmcB/OmcC. The individual roles of OmbB, OmaB and OmcB in extracellular reduction of Fe(III, however, have remained either uninvestigated or controversial. Here, we showed that replacements of ombB, omaB, omcB and ombB-omaB with an antibiotic gene in the presence of ombC-omaC-omcC had no impact on reduction of Fe(III-citrate by G. sulfurreducens PCA. Disruption of ombB, omaB, omcB and ombB-omaB in the absence of ombC-omaC-omcC, however, severely impaired the bacterial ability to reduce Fe(III-citrate as well as ferrihydrite. These results unequivocally demonstrate an overlapping role of ombB-omaB-omcB and ombC-omaC-omcC in extracellular Fe(III reduction by G. sulfurreducens PCA. Involvement of both ombB-omaB-omcB and ombC-omaC-omcC in extracellular Fe(III reduction reflects the importance of these trans-outer membrane protein complexes in the physiology of this bacterium. Moreover, the kinetics of Fe(III-citrate and ferrihydrite reduction by these mutants in the absence of ombC-omaC-omcC were nearly identical, which suggests that absence of any protein subunit eliminates function of OmaB/OmbB/OmcB protein complex. Finally, orfS was found to have a negative impact on the extracellular reduction of Fe(III-citrate and ferrihydrite in G. sulfurreducens PCA probably by serving as a transcriptional repressor.

  9. Necessity of electrically conductive pili for methanogenesis with magnetite stimulation

    Directory of Open Access Journals (Sweden)

    Oumei Wang

    2018-03-01

    Full Text Available Background Magnetite-mediated direct interspecies electron transfer (DIET between Geobacter and Methanosarcina species is increasingly being invoked to explain magnetite stimulation of methane production in anaerobic soils and sediments. Although magnetite-mediated DIET has been documented in defined co-cultures reducing fumarate or nitrate as the electron acceptor, the effects of magnetite have only been inferred in methanogenic systems. Methods Concentrations of methane and organic acid were analysed with a gas chromatograph and high-performance liquid chromatography, respectively. The concentration of HCl-extractable Fe(II was determined by the ferrozine method. The association of the defined co-cultures of G. metallireducens and M. barkeri with magnetite was observed with transmission electron micrographs. Results Magnetite stimulated ethanol metabolism and methane production in defined co-cultures of G. metallireducens and M. barkeri; however, magnetite did not promote methane production in co-cultures initiated with a culture of G. metallireducens that could not produce electrically conductive pili (e-pili, unlike the conductive carbon materials that facilitate DIET in the absence of e-pili. Transmission electron microscopy revealed that G. metallireducens and M. barkeri were closely associated when magnetite was present, as previously observed in G. metallireducens/G. sulfurreducens co-cultures. These results show that magnetite can promote DIET between Geobacter and Methanosarcina species, but not as a substitute for e-pili, and probably functions to facilitate electron transfer from the e-pili to Methanosarcina. Conclusion In summary, the e-pili are necessary for the stimulation of not only G. metallireducens/G. sulfurreducens, but also methanogenic G. metallireducens/M. barkeri co-cultures with magnetite.

  10. Extracellular Palladium Nanoparticle Production using Geobacter sulfurreducens

    KAUST Repository

    Yates, Matthew D.; Cusick, Roland D.; Logan, Bruce E.

    2013-01-01

    verified that Pd(0) nanoparticles were predominantly in the EPS surrounding cells in H2-fed cultures, with only a small number of particles visible inside the cell. Separation of the cells and EPS by centrifugation allowed reuse of the cell suspensions

  11. Promoting Interspecies Electron Transfer with Biochar

    Science.gov (United States)

    Chen, Shanshan; Rotaru, Amelia-Elena; Shrestha, Pravin Malla; Malvankar, Nikhil S.; Liu, Fanghua; Fan, Wei; Nevin, Kelly P.; Lovley, Derek R.

    2014-01-01

    Biochar, a charcoal-like product of the incomplete combustion of organic materials, is an increasingly popular soil amendment designed to improve soil fertility. We investigated the possibility that biochar could promote direct interspecies electron transfer (DIET) in a manner similar to that previously reported for granular activated carbon (GAC). Although the biochars investigated were 1000 times less conductive than GAC, they stimulated DIET in co-cultures of Geobacter metallireducens with Geobacter sulfurreducens or Methanosarcina barkeri in which ethanol was the electron donor. Cells were attached to the biochar, yet not in close contact, suggesting that electrons were likely conducted through the biochar, rather than biological electrical connections. The finding that biochar can stimulate DIET may be an important consideration when amending soils with biochar and can help explain why biochar may enhance methane production from organic wastes under anaerobic conditions. PMID:24846283

  12. Anode microbial communities produced by changing from microbial fuel cell to microbial electrolysis cell operation using two different wastewaters

    KAUST Repository

    Kiely, Patrick D.; Cusick, Roland; Call, Douglas F.; Selembo, Priscilla A.; Regan, John M.; Logan, Bruce E.

    2011-01-01

    Conditions in microbial fuel cells (MFCs) differ from those in microbial electrolysis cells (MECs) due to the intrusion of oxygen through the cathode and the release of H2 gas into solution. Based on 16S rRNA gene clone libraries, anode communities in reactors fed acetic acid decreased in species richness and diversity, and increased in numbers of Geobacter sulfurreducens, when reactors were shifted from MFCs to MECs. With a complex source of organic matter (potato wastewater), the proportion of Geobacteraceae remained constant when MFCs were converted into MECs, but the percentage of clones belonging to G. sulfurreducens decreased and the percentage of G. metallireducens clones increased. A dairy manure wastewater-fed MFC produced little power, and had more diverse microbial communities, but did not generate current in an MEC. These results show changes in Geobacter species in response to the MEC environment and that higher species diversity is not correlated with current. © 2010 Elsevier Ltd.

  13. Syntrophic Growth via Quinone-Mediated Interspecies Electron Transfer

    Directory of Open Access Journals (Sweden)

    Jessica A Smith

    2015-02-01

    Full Text Available The mechanisms by which microbial species exchange electrons are of interest because interspecies electron transfer can expand the metabolic capabilities of microbial communities. Previous studies with the humic substance analog anthraquinone-2,6-disulfonate (AQDS suggested that quinone-mediated interspecies electron transfer (QUIET is feasible, but it was not determined if sufficient energy is available from QUIET to support the growth of both species. Furthermore, there have been no previous studies on the mechanisms for the oxidation of anthrahydroquinone-2,6-disulfonate (AHQDS. A co-culture of Geobacter metallireducens and Geobacter sulfurreducens metabolized ethanol with the reduction of fumarate much faster in the presence of AQDS, and there was an increase in cell protein. G. sulfurreducens was more abundant, consistent with G. sulfurreducens obtaining electrons from acetate that G. metallireducens produced from ethanol, as well as from AHQDS. Cocultures initiated with a citrate synthase-deficient strain of G. sulfurreducens that was unable to use acetate as an electron donor also metabolized ethanol with the reduction of fumarate and cell growth, but acetate accumulated over time. G. sulfurreducens and G. metallireducens were equally abundant in these co-cultures reflecting the inability of the citrate synthase-deficient strain of G. sulfurreducens to metabolize acetate. Evaluation of the mechanisms by which G. sulfurreducens accepts electrons from AHQDS demonstrated that a strain deficient in outer-surface c-type cytochromes that are required for AQDS reduction was as effective at QUIET as the wild-type strain. Deletion of additional genes previously implicated in extracellular electron transfer also had no impact on QUIET. These results demonstrate that QUIET can yield sufficient energy to support the growth of both syntrophic partners, but that the mechanisms by which electrons are derived from extracellular hydroquinones require

  14. Geobacter anodireducens sp. nov., an exoelectrogenic microbe in bioelectrochemical systems

    KAUST Repository

    Sun, D.; Wang, A.; Cheng, S.; Yates, M.; Logan, B. E.

    2014-01-01

    -spore-forming, non-fermentative and non-motile. Cells were short, curved rods (0.8-1.3 µm long and 0.3 µm in diameter). Growth of strain SD-1(T) was observed at 15-42 °C and pH 6.0-8.5, with optimal growth at 30-35 °C and pH 7. Analysis of 16S rRNA gene sequences

  15. Impact of natural organic matter coatings on the microbial reduction of iron oxides

    Science.gov (United States)

    Poggenburg, Christine; Mikutta, Robert; Schippers, Axel; Dohrmann, Reiner; Guggenberger, Georg

    2018-03-01

    Iron (Fe) oxyhydroxides are important constituents of the soil mineral phase known to stabilize organic matter (OM) under oxic conditions. In an anoxic milieu, however, these Fe-organic associations are exposed to microbial reduction, releasing OM into soil solution. At present, only few studies have addressed the influence of adsorbed natural OM (NOM) on the reductive dissolution of Fe oxyhydroxides. This study therefore examined the impact of both the composition and concentration of adsorbed NOM on microbial Fe reduction with regard to (i) electron shuttling, (ii) complexation of Fe(II,III), (iii) surface site coverage and/or pore blockage, and (iv) aggregation. Adsorption complexes with varying carbon loadings were synthesized using different Fe oxyhydroxides (ferrihydrite, lepidocrocite, goethite, hematite, magnetite) and NOM of different origin (extracellular polymeric substances from Bacillus subtilis, OM extracted from soil Oi and Oa horizons). The adsorption complexes were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), N2 gas adsorption, electrophoretic mobility and particle size measurements, and OM desorption. Incubation experiments under anaerobic conditions were conducted for 16 days comparing two different strains of dissimilatory Fe(III)-reducing bacteria (Shewanella putrefaciens, Geobacter metallireducens). Mineral transformation during reduction was assessed via XRD and FTIR. Microbial reduction of the pure Fe oxyhydroxides was controlled by the specific surface area (SSA) and solubility of the minerals. For Shewanella putrefaciens, the Fe reduction of adsorption complexes strongly correlated with the concentration of potentially usable electron-shuttling molecules for NOM concentrations <2 mg C L-1, whereas for Geobacter metallireducens, Fe reduction depended on the particle size and thus aggregation of the adsorption complexes. These diverging results suggest that

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

    KAUST Repository

    Zhu, Xiuping; Yates, Matthew D.; Logan, Bruce E.

    2012-01-01

    larger range of set potentials was used to acclimate electroactive biofilms. The potentials of oxidation peaks obtained with G. sulfurreducens biofilms acclimated at 0.60 V (vs. Ag/AgCl) were different from those that developed at - 0.46 V, and both

  17. Long-range Electron Transport in Geobacter sulfurreducens Biofilms is Redox Gradient-Driven

    Science.gov (United States)

    2012-01-01

    modified version of the Nernst Equation (Eq. 2): ðXOxÞz=0; j = exp g nF RT Ej − Eo′ avg 1+ exp g nF RT Ej − Eo′ avg : [2] j= 1...www.pnas.org/cgi/doi/10.1073/pnas.1209829109 Snider et al. that follows directly from equation 21 in the work by Strycharz- Glaven et al. (14, 63

  18. Molecular analyis of rates of metal reductions and metabolic state of Geobacter species

    International Nuclear Information System (INIS)

    Lovley, Derek R.

    2008-01-01

    This project began with the simple goal of trying to understand the diversity of dissimilatory metal-reducing microorganisms that might be found in subsurface environments. It ended with a sophisticated understanding not only of what microorganisms are important for metal reduction in uranium-contaminated subsurface environments, but also their physiological status during in situ uranium bioremediation. These findings have provided unprecedented insight into uranium bioremediation and the methods by which this process might be optimized. A brief summary of the major accomplishments of the project is given.

  19. Link between capacity for current production and syntrophic growth in Geobacter species

    DEFF Research Database (Denmark)

    Rotaru, Amelia-Elena; Woodard, Trevor; Nevin, Kelly

    2015-01-01

    -culture with Methanosarcina barkeri, which is capable of direct interspecies electron transfer (DIET), but not with Methanospirillium hungatei capable only of H2 or formate transfer. Conductive granular activated carbon (GAC) stimulated metabolism of the G. hydrogenophilus - M. barkeri co-culture, consistent with electron...

  20. Illumina sequencing data for MEC study on high biofilm conductivity in a Geobacter-enriched biofilm

    Data.gov (United States)

    U.S. Environmental Protection Agency — This study systematically assessed intracellular electron transfer (IET) and extracellular electron transfer (EET) kinetics with respect to anode potential (Eanode)...

  1. Probing Single- to Multi-Cell Level Charge Transport in Geobacter sulfurreducens DL-1

    Science.gov (United States)

    2013-11-08

    cell was housed in a Faraday cage , yielding a noise level of o40 fA. All experiments were performed with more than 20 independent measurements across...indicated by purple arrow in Fig. 3a) after recording the stable baseline. To minimize the effect of external voltages and non-invasively probe the

  2. Final Report Systems Level Analysis of the Function and Adaptive Responses of Methanogenic Consortia

    Energy Technology Data Exchange (ETDEWEB)

    Lovley, Derek R. [Univ. of Massachusetts, Amherst, MA (United States)

    2015-03-09

    The purpose of this research was to determine whether the syntrophic microbial associations that are central to the functioning of methane-producing terrestrial wetlands can be predictively modeled with coupled multi-species genome-scale metabolic models. Such models are important because methane is an important greenhouse gas and there is a need to predictively model how the methane-producing microbial communities will respond to environmental perturbations, such as global climate change. The research discovered that the most prodigious methane-producing microorganisms on earth participate in a previously unrecognized form of energy exchange. The methane-producers Methanosaeta and Methanosarcina forge biological electrical connections with other microbes in order to obtain electrons to reduce carbon dioxide to methane. This direct interspecies electron transfer (DIET) was demonstrated in complex microbial communities as well as in defined co-cultures. For example, metatranscriptomic analysis of gene expression in both natural communities and defined co-cultures demonstrated that Methanosaeta species highly expressed genes for the enzymes for the reduction of carbon dioxide to methane. Furthermore, Methanosaeta’s electron-donating partners highly expressed genes for the biological electrical connections known as microbial nanowires. A series of studies involving transcriptomics, genome resequencing, and analysis of the metabolism of a series of strains with targeted gene deletions, further elucidated the mechanisms and energetics of DIET in methane-producing co-cultures, as well as in a co-culture of Geobacter metallireducens and Geobacter sulfurreducens, which provided a system for studying DIET with two genetically tractable partners. Genome-scale modeling of DIET in the G. metallireducens/G. sulfurreducens co-culture suggested that DIET provides more energy to the electron-donating partner that electron exchange via interspecies hydrogen transfer, but that the

  3. A method for high throughput bioelectrochemical research based on small scale microbial electrolysis cells

    KAUST Repository

    Call, Douglas F.

    2011-07-01

    There is great interest in studying exoelectrogenic microorganisms, but existing methods can require expensive electrochemical equipment and specialized reactors. We developed a simple system for conducting high throughput bioelectrochemical research using multiple inexpensive microbial electrolysis cells (MECs) built with commercially available materials and operated using a single power source. MECs were small crimp top serum bottles (5mL) with a graphite plate anode (92m 2/m 3) and a cathode of stainless steel (SS) mesh (86m 2/m 3), graphite plate, SS wire, or platinum wire. The highest volumetric current density (240A/m 3, applied potential of 0.7V) was obtained using a SS mesh cathode and a wastewater inoculum (acetate electron donor). Parallel operated MECs (single power source) did not lead to differences in performance compared to non-parallel operated MECs, which can allow for high throughput reactor operation (>1000 reactors) using a single power supply. The utility of this method for cultivating exoelectrogenic microorganisms was demonstrated through comparison of buffer effects on pure (Geobacter sulfurreducens and Geobacter metallireducens) and mixed cultures. Mixed cultures produced current densities equal to or higher than pure cultures in the different media, and current densities for all cultures were higher using a 50mM phosphate buffer than a 30mM bicarbonate buffer. Only the mixed culture was capable of sustained current generation with a 200mM phosphate buffer. These results demonstrate the usefulness of this inexpensive method for conducting in-depth examinations of pure and mixed exoelectrogenic cultures. © 2011 Elsevier B.V.

  4. Nonproteinogenic D-amino acids at millimolar concentrations are a toxin for anaerobic microorganisms relevant to early Earth and other anoxic planets.

    Science.gov (United States)

    Nixon, Sophie L; Cockell, Charles S

    2015-03-01

    The delivery of extraterrestrial organics to early Earth provided a potentially important source of carbon and energy for microbial life. Optically active organic compounds of extraterrestrial origin exist in racemic form, yet life on Earth has almost exclusively selected for L- over D-enantiomers of amino acids. Although D-enantiomers of proteinogenic amino acids are known to inhibit aerobic microorganisms, the role of concentrated nonproteinogenic meteoritic D-amino acids on anaerobic metabolisms relevant to early Earth and other anoxic planets such as Mars is unknown. Here, we test the inhibitory effect of D-enantiomers of two nonproteinogenic amino acids common to carbonaceous chondrites, norvaline and α-aminobutyric acid, on microbial iron reduction. Three pure strains (Geobacter bemidjiensis, Geobacter metallireducens, Geopsychrobacter electrodiphilus) and an iron-reducing enrichment culture were grown in the presence of 10 mM D-enantiomers of both amino acids. Further tests were conducted to assess the inhibitory effect of these D-amino acids at 1 and 0.1 mM. The presence of 10 mM D-norvaline and D-α-aminobutyric acid inhibited microbial iron reduction by all pure strains and the enrichment. G. bemidjiensis was not inhibited by either amino acid at 0.1 mM, but D-α-aminobutyric acid still inhibited at 1 mM. Calculations using published meteorite accumulation rates to the martian surface indicate D-α-aminobutyric acid may have reached inhibitory concentrations in little over 1000 years during peak infall. These data show that, on a young anoxic planet, the use of one enantiomer over another may render the nonbiological enantiomer an environmental toxin. Processes that generate racemic amino acids in the environment, such as meteoritic infall or impact synthesis, would have been toxic processes and could have been a selection pressure for the evolution of early racemases.

  5. Iron and manganese in anaerobic respiration: environmental significance, physiology, and regulation

    Science.gov (United States)

    Nealson, K. H.; Saffarini, D.

    1994-01-01

    Dissimilatory iron and/or manganese reduction is known to occur in several organisms, including anaerobic sulfur-reducing organisms such as Geobacter metallireducens or Desulfuromonas acetoxidans, and facultative aerobes such as Shewanella putrefaciens. These bacteria couple both carbon oxidation and growth to the reduction of these metals, and inhibitor and competition experiments suggest that Mn(IV) and Fe(III) are efficient electron acceptors similar to nitrate in redox abilities and capable of out-competing electron acceptors of lower potential, such as sulfate (sulfate reduction) or CO2 (methanogenesis). Field studies of iron and/or manganese reduction suggest that organisms with such metabolic abilities play important roles in coupling the oxidation of organic carbon to metal reduction under anaerobic conditions. Because both iron and manganese oxides are solids or colloids, they tend to settle downward in aquatic environments, providing a physical mechanism for the movement of oxidizing potential into anoxic zones. The resulting biogeochemical metal cycles have a strong impact on many other elements including carbon, sulfur, phosphorous, and trace metals.

  6. Electrochemical and genomic analysis of novel electroactive isolates obtained via potentiostatic enrichment from tropical sediment

    Science.gov (United States)

    Doyle, Lucinda E.; Yung, Pui Yi; Mitra, Sumitra D.; Wuertz, Stefan; Williams, Rohan B. H.; Lauro, Federico M.; Marsili, Enrico

    2017-07-01

    Enrichment of electrochemically-active microorganisms (EAM) to date has mostly relied on microbial fuel cells fed with wastewater. This study aims to enrich novel EAM by exposing tropical sediment, not frequently reported in the literature, to sustained anodic potentials. Voltamperometric techniques and electrochemical impedance spectroscopy, performed over a wide range of potentials, characterise extracellular electron transfer (EET) over time. Applied potential is found to affect biofilm electrochemical signature. Geobacter metallireducens is heavily enriched on the electrodes, as determined by metagenomic and metatranscriptomic analysis, in the first report of the species in a lactate-fed system. Two novel isolates are grown in pure culture from the enrichment, identified by 16S rRNA gene sequencing as Aeromonas and Enterobacter, respectively. The names proposed are Aeromonas sp. CL-1 and Enterobacter sp. EA-1. Both isolates are capable of EET on carbon felt and screen-printed carbon electrodes without the addition of exogenous redox mediators. Enterobacter sp. EA-1 can also perform mediated electron transfer using the soluble redox mediator 2-hydroxy-1,4-naphthoquinone (HNQ). Both isolates are able to use acetate and lactate as electron donors. This work outlines a comprehensive methodology for characterising novel EAM from unconventional inocula.

  7. Use of Novel Reinforced Cation Exchange Membranes for Microbial Fuel Cells

    International Nuclear Information System (INIS)

    Kamaraj, Sathish-Kumar; Romano, Sergio Mollá; Moreno, Vicente Compañ; Poggi-Varaldo, H.M.; Solorza-Feria, O.

    2015-01-01

    This work has been focused on the synthesis and characterization of different blended membranes SPEEK-35PVA (Water), SPEEK-35PVA (DMAc) prepared by casting and nanofiber-reinforced proton exchange membranes Nafion-PVA-15, Nafion-PVA-23 and SPEEK/PVA-PVB. The two first reinforced membranes were made up of Nafion® polymer deposited between polyvinyl alcohol (PVA) nanofibers. The last composite membrane is considered because the PVA is a hydrophilic polymer which forms homogeneous blends with SPEEK suitable to obtain high proton conductivity, while the hydrophobic PVB can produce blends in a phase separation morphology in which very low water uptake can be found. The synthesized membranes showed an outstanding stability, high proton conductivity, and enhanced mechanical and barrier properties. The membranes were characterized in single chamber microbial fuel cells (SCMFCs) using electrochemically enriched high sodic saline hybrid H-inocula (Geobacter metallireducen, Desulfurivibrio alkaliphilus, and Marinobacter adhaerens) as biocatalyst. The best performance was obtained with Nafion-PVA-15 membrane, which achieved a maximum power density of 1053 mW/m 3 at a cell voltage of 340 mV and displayed the lowest total internal resistance (Rint ≈ 522 Ω). This result is in agreement with the low oxygen permeability and the moderate conductivity found in this kind of membranes. These results are encouraging towards obtaining high concentrated sodic saline model wastewater exploiting MFCs

  8. Salt removal using multiple microbial desalination cells under continuous flow conditions

    KAUST Repository

    Qu, Youpeng

    2013-05-01

    Four microbial desalination cells (MDCs) were hydraulically connected and operated under continuous flow conditions. The anode solution from the first MDC flowed into the cathode, and then on to the anode of the next reactor, which avoided pH imbalances that inhibit bacterial metabolism. The salt solution also moved through each desalination chamber in series. Increasing the hydraulic retention times (HRTs) of the salt solution from 1 to 2. days increased total NaCl removal from 76 ± 1% to 97 ± 1%, but coulombic efficiencies decreased from 49 ± 4% to 35 ± 1%. Total COD removals were similar at both HRTs (60 ± 2%, 2. days; 59 ± 2%, 1. day). Community analysis of the anode biofilms showed that bacteria most similar to the xylose fermenting bacterium Klebsiella ornithinolytica predominated in the anode communities, and sequences most similar to Geobacter metallireducens were identified in all MDCs except the first one. These results demonstrated successful operation of a series of hydraulically connected MDCs and good desalination rates. © 2013 Elsevier B.V..

  9. Inocula selection in microbial fuel cells based on anodic biofilm abundance of Geobacter sulfurreducens

    DEFF Research Database (Denmark)

    Sun, Guotao; Rodrigues, Diogo De Sacadura; Thygesen, Anders

    2016-01-01

    with naturally occurring mixed inocula. In this study, the electrochemical performance of MFCs and microbial community evolution were evaluated for three inocula including domestic wastewater (DW), lake sediment (LS) and biogas sludge (BS) with varying substrate loading (Lsub) and external resistance (Rext....... The data obtained contribute to understanding the microbial community response to Lsub and Rext for optimizing electricity generation in MFCs....

  10. Mechanisms for Fe(III) oxide reduction in sedimentary environments

    Science.gov (United States)

    Nevin, Kelly P.; Lovely, Derek R.

    2002-01-01

    Although it was previously considered that Fe(III)-reducing microorganisms must come into direct contact with Fe(III) oxides in order to reduce them, recent studies have suggested that electron-shuttling compounds and/or Fe(III) chelators, either naturally present or produced by the Fe(III)-reducing microorganisms themselves, may alleviate the need for the Fe(III) reducers to establish direct contact with Fe(III) oxides. Studies with Shewanella alga strain BrY and Fe(III) oxides sequestered within microporous beads demonstrated for the first time that this organism releases a compound(s) that permits electron transfer to Fe(III) oxides which the organism cannot directly contact. Furthermore, as much as 450 w M dissolved Fe(III) was detected in cultures of S. alga growing in Fe(III) oxide medium, suggesting that this organism releases compounds that can solublize Fe(III) from Fe(III) oxide. These results contrast with previous studies, which demonstrated that Geobacter metallireducens does not produce electron-shuttles or Fe(III) chelators. Some freshwater aquatic sediments and groundwaters contained compounds, which could act as electron shuttles by accepting electrons from G. metallireducens and then transferring the electrons to Fe(III). However, other samples lacked significant electron-shuttling capacity. Spectroscopic studies indicated that the electron-shuttling capacity of the waters was not only associated with the presence of humic substances, but water extracts of walnut, oak, and maple leaves contained electron-shuttling compounds did not appear to be humic substances. Porewater from a freshwater aquatic sediment and groundwater from a petroleum-contaminated aquifer contained dissolved Fe(III) (4-16 w M), suggesting that soluble Fe(III) may be available as an electron acceptor in some sedimentary environments. These results demonstrate that in order to accurately model the mechanisms for Fe(III) reduction in sedimentary environments it will be necessary

  11. Sediment studies of the biological factors controlling the reduction of U(VI)

    International Nuclear Information System (INIS)

    Lovley, Derek R.

    2004-01-01

    Studies were conducted primarily with sediments, both in laboratory incubations and in a field experiment, with supporting studies with pure cultures. To our knowledge the sediment studies were the first on microbial U(VI) reduction in actual uranium-contaminated subsurface sediments, under conditions that mimic those found in situ. Important findings included: (1) U(VI) reduction is a biotic process in subsurface sediments. (2) U(VI) reduction can be stimulated most effectively with the addition of acetate. Although it had been speculated that microbial U(VI) reduction might be capable of this type of environmental remediation ever since the discovery of microbial U(VI) reduction, this had not been previously demonstrated under environmentally relevant conditions. (3) U(VI) is reduced concurrently with Fe(III) and prior to sulfate reduction. U(VI) and Fe(III) reduction proceeded concurrently, accompanied by a dramatic enrichment in organisms in the Geobacteraceae. Sulfate-reducing microorganisms do not appear to be important components of the microbial community reducing U(VI) in these subsurface sediments. (4) Nitrate has important influences on U(VI) reduction. Nitrate inhibits the reduction of metals until nitrate is depleted. Fe(III)-reducing microorganisms such as Geobacter metallireducens and Desulfitobacterium species can oxidize Fe(II) with the reduction of nitrate which is an important consideration because our previous studies have demonstrated that freshly precipitated Fe(III) oxides can reoxidize U(IV) to U(VI). The discovery that G. metallireducens can ''run backwards'' and oxidize U(IV) when nitrate is present reveals another mechanism preventing precipitation of U(IV) in the presence of nitrate as well as potential novel strategy for removing uranium from the subsurface after a site has been remediated. (5) Importance of understanding Fe(III) forms available for microbial reduction. Fe(III) is orders of magnitude more abundant than U(VI) as an

  12. In-Situ Survival Mechanisms of U and Tc Reducing Bacteria in Contaminated Sediments

    International Nuclear Information System (INIS)

    Krumholz, Lee R.

    2005-01-01

    periplasmic space and outer membrane associated proteins. Through blast search analysis, we also showed that 81 out of 94 proteins shown to be important in sediment survival have homologs in D. vulgaris, 70 have homologs in Geobacter metallireducens, and 69 have homologs in Geobacter sulfurreducens PCA. Some interesting proteins include ribonucleotide reductase and chemotaxis related proteins. Ribonucleotide reductase catalyzes the reductive synthesis of deoxyribonucleotides from their corresponding ribonucleotides, providing the precursors necessary for DNA synthesis. Two ribonucleotide reductase genes (nrdE, nrdD) were found to be essential for G20 survival in the sediment, but not essential for growth in the lactate-sulfate medium. Bacterial methyl-accepting chemotaxis proteins (MCP) respond to changes in the concentration of attractants and repellents in the environment

  13. Biological Recovery of Platinum Complexes from Diluted Aqueous Streams by Axenic Cultures.

    Directory of Open Access Journals (Sweden)

    Synthia Maes

    Full Text Available The widespread use of platinum in high-tech and catalytic applications has led to the production of diverse Pt loaded wastewaters. Effective recovery strategies are needed for the treatment of low concentrated waste streams to prevent pollution and to stimulate recovery of this precious resource. The biological recovery of five common environmental Pt-complexes was studied under acidic conditions; the chloro-complexes PtCl42- and PtCl62-, the amine-complex Pt(NH34Cl2 and the pharmaceutical complexes cisplatin and carboplatin. Five bacterial species were screened on their platinum recovery potential; the Gram-negative species Shewanella oneidensis MR-1, Cupriavidus metallidurans CH34, Geobacter metallireducens, and Pseudomonas stutzeri, and the Gram-positive species Bacillus toyonensis. Overall, PtCl42- and PtCl62- were completely recovered by all bacterial species while only S. oneidensis and C. metallidurans were able to recover cisplatin quantitatively (99%, all in the presence of H2 as electron donor at pH 2. Carboplatin was only partly recovered (max. 25% at pH 7, whereas no recovery was observed in the case of the Pt-tetraamine complex. Transmission electron microscopy (TEM revealed the presence of both intra- and extracellular platinum particles. Flow cytometry based microbial viability assessment demonstrated the decrease in number of intact bacterial cells during platinum reduction and indicated C. metallidurans to be the most resistant species. This study showed the effective and complete biological recovery of three common Pt-complexes, and estimated the fate and transport of the Pt-complexes in wastewater treatment plants and the natural environment.

  14. Application of a tetrazolium dye as an indicator of viability in anaerobic bacteria.

    Science.gov (United States)

    Bhupathiraju, V K; Hernandez, M; Landfear, D; Alvarez-Cohen, L

    1999-09-01

    The use of the redox dye 5-cyano-2,3,-ditolyl tetrazolium chloride (CTC) for evaluating the metabolic activity of aerobic bacteria has gained wide application in recent years. In this study, we examined the utility of CTC in capturing the metabolic activity of anaerobic bacteria. In addition, the factors contributing to abiotic reduction of CTC were also examined. CTC was used in conjunction with the fluorochrome 5-(4,6-dichlorotriazinyl) aminofluorescein (DTAF), that targets bacterial cell wall proteins, to quantitate the active fraction of total bacterial numbers. Facultative anaerobic bacteria, including Escherichia coli grown fermentatively, and Pseudomonas chlorophis, P. fluorescens, P. stutzeri, and P. pseudoalcalegenes subsp. pseudoalcalegenes grown under nitrate-reducing conditions, actively reduced CTC during all phases of growth. Greater than 95% of these cells accumulated intracellular CTC-formazan crystals during the exponential phase. Obligate anaerobic bacteria, including Syntrophus aciditrophicus grown fermentatively, Geobacter sulfurreducens grown with fumarate as the electron acceptor, Desulfovibrio desulfuricans subsp. desulfuricans and D. halophilus grown under sulfate-reducing conditions, Methanobacterium formicicum grown on formate, H2 and CO2, and Methanobacterium thermoautotrophicum grown autotrophically on H2 and CO2 all reduced CTC to intracellular CTC-formazan crystals. The optimal CTC concentration for all organisms examined was 5 mM. Anaerobic CTC incubations were not required for quantification of anaerobically grown cells. CTC-formazan production by all cultures examined was proportional to biomass production, and CTC reduction was observed even in the absence of added nutrients. CTC was reduced by culture fluids containing ferric citrate as electron acceptor following growth of either G. metallireducens or G. sulfurreducens. Abiotic reduction of CTC was observed in the presence of ascorbic acid, cysteine hydrochloride, dithiothreitol

  15. Novel processes for anaerobic sulfate production from elemental sulfur by sulfate-reducing bacteria

    Science.gov (United States)

    Lovley, D.R.; Phillips, E.J.P.

    1994-01-01

    Sulfate reducers and related organisms which had previously been found to reduce Fe(III) with H2 or organic electron donors oxidized S0 to sulfate when Mn(IV) was provided as an electron acceptor. Organisms catalyzing this reaction in washed cell suspensions included Desulfovibrio desulfuricans, Desulfomicrobium baculatum. Desulfobacterium autotrophicum, Desulfuromonas acetoxidans, and Geobacter metallireducens. These organisms produced little or no sulfate from S0 with Fe(III) as a potential electron acceptor or in the absence of an electron acceptor. In detailed studies with Desulfovibrio desulfuricans, the stoichiometry of sulfate and Mn(II) production was consistent with the reaction S0 + 3 MnO2 + 4H+ ???SO42- + 3Mn(II) + 2H2O. None of the organisms evaluated could be grown with S0 as the sole electron donor and Mn(IV) as the electron acceptor. In contrast to the other sulfate reducers evaluated, Desulfobulbus propionicus produced sulfate from S0 in the absence of an electron acceptor and Fe(III) oxide stimulated sulfate production. Sulfide also accumulated in the absence of Mn(IV) or Fe(III). The stoichiometry of sulfate and sulfide production indicated that Desulfobulbus propionicus disproportionates S0 as follows: 4S0 + 4H2O???SO42- + 3HS- + 5 H+. Growth of Desulfobulbus propionicus with S0 as the electron donor and Fe(III) as a sulfide sink and/or electron acceptor was very slow. The S0 oxidation coupled to Mn(IV) reduction described here provides a potential explanation for the Mn(IV)-dependent sulfate production that previous studies have observed in anoxic marine sediments. Desulfobulbus propionicus is the first example of a pure culture known to disproportionate S0.

  16. STRUCTURE AND FUNCTION OF SUBSURFACE MICROBIAL COMMUNITIES AFFECTING RADIONUCLIDE TRANSPORT AND BIOIMMOBILIZATION

    Energy Technology Data Exchange (ETDEWEB)

    Joel E. Kostka; Lee Kerkhof; Kuk-Jeong Chin; Martin Keller; Joseph W. Stucki

    2011-06-15

    (comprising 50% to 100% of rRNA detected). (2) We demonstrated for the first time that the function of microbial communities can be quantified in subsurface sediments using messenger RNA assays (molecular proxies) under in situ conditions. (3) Active Geobacteraceae were identified and phylogenetically characterized from the cDNA of messenger RNA extracted from ORFRC subsurface sediment cores. Multiple clone sequences were retrieved from G. uraniireducens, G. daltonii, and G. metallireducens. (4) Results show that Geobacter strain FRC-32 is capable of growth on benzoate, toluene and benzene as the electron donor, thereby providing evidence that this strain is physiologically distinct from other described members of the subsurface Geobacter clade. (5) Fe(III)-reducing bacteria transform structural Fe in clay minerals from their layer edges rather than from their basal surfaces.

  17. Microbial Fe(III) Oxide Reduction in Chocolate Pots Hot Springs, Yellowstone National Park

    Science.gov (United States)

    Fortney, N. W.; Roden, E. E.; Boyd, E. S.; Converse, B. J.

    2014-12-01

    Previous work on dissimilatory iron reduction (DIR) in Yellowstone National Park (YNP) has focused on high temperature, low pH environments where soluble Fe(III) is utilized as an electron acceptor for respiration. Much less attention has been paid to DIR in lower temperature, circumneutral pH environments, where solid phase Fe(III) oxides are the dominant forms of Fe(III). This study explored the potential for DIR in the warm (ca. 40-50°C), circumneutral pH Chocolate Pots hot springs (CP) in YNP. Most probable number (MPN) enumerations and enrichment culture studies confirmed the presence of endogenous microbial communities that reduced native CP Fe(III) oxides. Enrichment cultures demonstrated sustained DIR coupled to acetate and lactate oxidation through repeated transfers over ca. 450 days. Pyrosequencing of 16S rRNA genes indicated that the dominant organisms in the enrichments were closely affiliated with the well known Fe(III) reducer Geobacter metallireducens. Additional taxa included relatives of sulfate reducing bacterial genera Desulfohalobium and Thermodesulfovibrio; however, amendment of enrichments with molybdate, an inhibitor of sulfate reduction, suggested that sulfate reduction was not a primary metabolic pathway involved in DIR in the cultures. A metagenomic analysis of enrichment cultures is underway in anticipation of identifying genes involved in DIR in the less well-characterized dominant organisms. Current studies are aimed at interrogating the in situ microbial community at CP. Core samples were collected along the flow path (Fig. 1) and subdivided into 1 cm depth intervals for geochemical and microbiological analysis. The presence of significant quantities of Fe(II) in the solids indicated that DIR is active in situ. A parallel study investigated in vitro microbial DIR in sediments collected from three of the coring sites. DNA was extracted from samples from both studies for 16S rRNA gene and metagenomic sequencing in order to obtain a

  18. Diverse anaerobic Cr(VI) tolerant bacteria from Cr(VI)-contaminated 100H site at Hanford

    Science.gov (United States)

    Chakraborty, R.; Phan, R.; Lam, S.; Leung, C.; Brodie, E. L.; Hazen, T. C.

    2007-12-01

    Hexavalent Chromium [Cr(VI)] is a widespread contaminant found in soil, sediment, and ground water. Cr(VI) is more soluble, toxic, carcinogenic, and mutagenic compared to its reduced form Cr(III). In order to stimulate microbially mediated reduction of Cr(VI), a poly-lactate compound HRC was injected into the chromium contaminated aquifers at site 100H at Hanford. Based on the results of the bacterial community composition using high-density DNA microarray analysis of 16S rRNA gene products, we recently investigated the diversity of the dominant anaerobic culturable microbial population present at this site and their role in Cr(VI) reduction. Positive enrichments set up at 30°C using specific defined anaerobic media resulted in the isolation of an iron reducing isolate strain HAF, a sulfate reducing isolate strain HBLS and a nitrate reducing isolate, strain HLN among several others. Preliminary 16S rDNA sequence analysis identifies strain HAF as Geobacter metallireducens, strain HLN as Pseudomonas stutzeri and strain HBLS as a member of Desulfovibrio species. Strain HAF isolated with acetate as the electron donor utilized propionate, glycerol and pyruvate as alternative carbon sources, and reduced metals like Mn(IV) and Cr(VI). Growth was optimal at 37°C, pH of 6.5 and 0% salinity. Strain HLN isolated with lactate as electron donor utilized acetate, glycerol and pyruvate as alternative carbon sources, and reduced metals like Mn(IV) and Cr(VI). Optimal growth was observed at 37°C, at a pH of 7.5 and 0.3% salinity. Anaerobic active washed cell suspension of strain HLN reduced almost 95 micromolar Cr(VI) within 4 hours relative to controls. Further, with 100 micromolar Cr(VI) as the sole electron acceptor, cells of strain HLN grew to cell numbers of 4.05X 107/ml over a period of 24hrs after an initial lag, demonstrating direct enzymatic Cr(VI) reduction by this species. 10mM lactate served as the sole electron donor. These results demonstrate that Cr

  19. 6-Oxocyclohex-1-ene-1-carbonyl-coenzyme A hydrolases from obligately anaerobic bacteria: characterization and identification of its gene as a functional marker for aromatic compounds degrading anaerobes.

    Science.gov (United States)

    Kuntze, Kevin; Shinoda, Yoshifumi; Moutakki, Housna; McInerney, Michael J; Vogt, Carsten; Richnow, Hans-Hermann; Boll, Matthias

    2008-06-01

    In anaerobic bacteria, most aromatic growth substrates are channelled into the benzoyl-coenzyme A (CoA) degradation pathway where the aromatic ring is dearomatized and cleaved into an aliphatic thiol ester. The initial step of this pathway is catalysed by dearomatizing benzoyl-CoA reductases yielding the two electron-reduction product, cyclohexa-1,5-diene-1-carbonyl-CoA, to which water is subsequently added by a hydratase. The next two steps have so far only been studied in facultative anaerobes and comprise the oxidation of the 6-hydroxyl-group to 6-oxocyclohex-1-ene-1-carbonyl-CoA (6-OCH-CoA), the addition of water and hydrolytic ring cleavage yielding 3-hydroxypimelyl-CoA. In this work, two benzoate-induced genes from the obligately anaerobic bacteria, Geobacter metallireducens (bamA(Geo)) and Syntrophus aciditrophicus (bamA(Syn)), were heterologously expressed in Escherichia coli, purified and characterized as 6-OCH-CoA hydrolases. Both enzymes consisted of a single 43 kDa subunit. Some properties of the enzymes are presented and compared with homologues from facultative anaerobes. An alignment of the nucleotide sequences of bamA(Geo) and bamA(Syn) with the corresponding genes from facultative anaerobes identified highly conserved DNA regions, which enabled the discrimination of genes coding for 6-OCH-CoA hydrolases from those coding for related enzymes. A degenerate oligonucleotide primer pair was deduced from conserved regions and applied in polymerase chain reaction reactions. Using these primers, the expected DNA fragment of the 6-OCH-CoA hydrolase genes was specifically amplified from the DNA of nearly all known facultative and obligate anaerobes that use aromatic growth substrates. The only exception was the aromatic compound-degrading Rhodopseudomonas palustris, which uniquely uses a modified benzoyl-CoA degradation pathway. Using the oligonucleotide primers, the expected DNA fragment was also amplified in a toluene-degrading and a m

  20. Impact of fulvic acids on bio-methanogenic treatment of municipal solid waste incineration leachate.

    Science.gov (United States)

    Dang, Yan; Lei, Yuqing; Liu, Zhao; Xue, Yiting; Sun, Dezhi; Wang, Li-Ying; Holmes, Dawn E

    2016-12-01

    A considerable amount of leachate with high fulvic acid (FA) content is generated during the municipal solid waste (MSW) incineration process. This incineration leachate is usually processed by downstream bio-methanogenic treatment. However, few studies have examined the impact that these compounds have on methanogenesis and how they are degraded and transformed during the treatment process. In this study, a laboratory-scale expanded granular sludge bed (EGSB) reactor was operated with MSW incineration leachate containing various concentrations of FA (1500 mg/L to 8000 mg/L) provided as the influent. We found that FA degradation rates decreased from 86% to 72% when FA concentrations in the reactor were increased, and that molecular size, level of humification and aromatization of the residual FA macromolecules all increased after bio-methanogenic treatment. Increasing FA influent concentrations also inhibited growth of hydrogenotrophic methanogens from the genus Methanobacterium and syntrophic bacteria from the genus Syntrophomonas, which resulted in a decrease in methane production and a concomitant increase in CO 2 content in the biogas. Sequences most similar to species from the genus Anaerolinea went up as FA concentrations increased. Bacteria from this genus are capable of extracellular electron transfer and may be using FA as an electron acceptor for growth or as a shuttle for syntrophic exchange with other microorganisms in the reactor. In order to determine whether FA could serve as an electron shuttle to promote syntrophy in an anaerobic digester, co-cultures of Geobacter metallireducens and G. sulfurreducens were grown in the presence of FA from raw leachate or from residual bioreactor effluent. While raw FA stimulated electron transfer between these two bacteria, residual FA did not have any electron shuttling abilities, indicating that FA underwent a significant transformation during the bio-methanogenic treatment process. These results are

  1. Assessment of Bacterial Degradation of Aromatic Hydrocarbons in the Environment by Analysis of Stable Carbon Isotope Fractionation

    Energy Technology Data Exchange (ETDEWEB)

    Meckenstock, Rainer U. [Eberhard-Karls University of Tuebingen, Center for Applied Geoscience (Germany)], E-mail: rainer.meckenstock@uni-tuebingen.de; Morasch, Barbara [University of Konstanz, Faculty of Biology (Germany); Kaestner, Matthias; Vieth, Andrea; Richnow, Hans Hermann [Center for Environmental Research, Department of Remediation Research (Germany)

    2002-05-15

    {sup 13}C/{sup 12}C stable carbon isotope fractionation was used to assess biodegradation in contaminated aquifers with toluene as a model compound. Different strains of anaerobic bacteria (Thauera aromatica, Geobacter metallireducens, and the sulfate-reducing strain TRM1) showed consistent {sup 13}C/{sup 12}C carbon isotope fractionation with fractionation factors between {alpha}C = 1.0017 and 1.0018. In contrast, three cultures of aerobic organisms, using different mono- and dioxygenase enzyme systems to initiate toluene degradation, showed variable isotope fractionation factors of {alpha}C = 1.0027 (Pseudomonasputida strain mt-2), {alpha}C = 1.0011 (Ralstonia picketii), and{alpha}C = 1.0004 (Pseudomonas putida strain F1). The great variability of isotope fractionation between different aerobic bacterial strains suggests that interpretation of isotope data in oxic habitats can only be qualitative. A soil column was run as a model system for contaminated aquifers with toluene as the carbon source and sulfate as the electron acceptor and samples were taken at different ports along the column. Microbial toluene degradation was calculated based on the {sup 13}C/{sup 12}C isotope fractionation factors of the batch culture experiments together with the observed {sup 13}C/{sup 12}C isotope shifts of the residual toluene fractions. The calculated percentage of biodegradation, B, correlated well with the decreasing toluene concentrations at the sampling ports and indicated the increasing extent of biodegradation along the column. The theoretical toluene concentrations as calculated based on the isotope values matched the measured concentrations at the different sampling ports indicating that the Rayleigh equation can be used to calculate biodegradation in quasi closed systems based on measured isotope shifts. A similar attempt was performed to assess toluene degradation in a contaminated, anoxic aquifer. A transect of groundwater wells was monitored along the main

  2. Assessment of Bacterial Degradation of Aromatic Hydrocarbons in the Environment by Analysis of Stable Carbon Isotope Fractionation

    International Nuclear Information System (INIS)

    Meckenstock, Rainer U.; Morasch, Barbara; Kaestner, Matthias; Vieth, Andrea; Richnow, Hans Hermann

    2002-01-01

    13 C/ 12 C stable carbon isotope fractionation was used to assess biodegradation in contaminated aquifers with toluene as a model compound. Different strains of anaerobic bacteria (Thauera aromatica, Geobacter metallireducens, and the sulfate-reducing strain TRM1) showed consistent 13 C/ 12 C carbon isotope fractionation with fractionation factors between αC = 1.0017 and 1.0018. In contrast, three cultures of aerobic organisms, using different mono- and dioxygenase enzyme systems to initiate toluene degradation, showed variable isotope fractionation factors of αC = 1.0027 (Pseudomonasputida strain mt-2), αC = 1.0011 (Ralstonia picketii), andαC = 1.0004 (Pseudomonas putida strain F1). The great variability of isotope fractionation between different aerobic bacterial strains suggests that interpretation of isotope data in oxic habitats can only be qualitative. A soil column was run as a model system for contaminated aquifers with toluene as the carbon source and sulfate as the electron acceptor and samples were taken at different ports along the column. Microbial toluene degradation was calculated based on the 13 C/ 12 C isotope fractionation factors of the batch culture experiments together with the observed 13 C/ 12 C isotope shifts of the residual toluene fractions. The calculated percentage of biodegradation, B, correlated well with the decreasing toluene concentrations at the sampling ports and indicated the increasing extent of biodegradation along the column. The theoretical toluene concentrations as calculated based on the isotope values matched the measured concentrations at the different sampling ports indicating that the Rayleigh equation can be used to calculate biodegradation in quasi closed systems based on measured isotope shifts. A similar attempt was performed to assess toluene degradation in a contaminated, anoxic aquifer. A transect of groundwater wells was monitored along the main direction of the groundwater flow and revealed decreasing

  3. Nitrate Enhanced Microbial Cr(VI) Reduction-Final Report

    Energy Technology Data Exchange (ETDEWEB)

    John F. Stolz

    2011-06-15

    A major challenge for the bioremediation of radionuclides (i.e., uranium, technetium) and metals (i.e., Cr(VI), Hg) is the co-occurrence of nitrate as it can inhibit metal transformation. Denitrification (nitrate reduction to dinitrogen gas) is considered the most important ecological process. For many metal and metalloid reducing bacteria, however, ammonia is the end product through respiratory nitrate reduction (RNRA). The focus of this work was to determine how RNRA impacts Cr(VI) transformation. The goal was to elucidate the specific mechanism(s) that limits Cr(VI) reduction in the presence of nitrate and to use this information to develop strategies that enhance Cr(VI) reduction (and thus detoxification). Our central hypothesis is that nitrate impacts the biotransformation of metals and metalloids in three ways 1) as a competitive alternative electron acceptor (inhibiting transformation), 2) as a co-metabolite (i.e., concomitant reduction, stimulating transformation), and 3) as an inducer of specific proteins and pathways involved in oxidation/reduction reactions (stimulating transformation). We have identified three model organisms, Geobacter metallireducens (mechanism 1), Sulfurospirillum barnesii, (mechasism 2), and Desulfovibrio desulfuricans (mechanisms 3). Our specific aims were to 1) investigate the role of Cr(VI) concentration on the kinetics of both growth and reduction of nitrate, nitrite, and Cr(VI) in these three organisms; 2) develop a profile of bacterial enzymes involved in nitrate transformation (e.g., oxidoreductases) using a proteomic approach; 3) investigate the function of periplasmic nitrite reductase (Nrf) as a chromate reductase; and 4) develop a strategy to maximize microbial chromium reduction in the presence of nitrate. We found that growth on nitrate by G. metallireducens was inhibited by Cr(VI). Over 240 proteins were identified by LC/MS-MS. Redox active proteins, outer membrane heavy metal efflux proteins, and chemotaxis sensory

  4. Characterization of phenols biodegradation by compound specific stable isotope analysis

    Science.gov (United States)

    Wei, Xi; Gilevska, Tetyana; Wenzig, Felix; Hans, Richnow; Vogt, Carsten

    2015-04-01

    Biodegradation of phenol and alkylphenols has been described under both oxic and anoxic conditions. In the absence of molecular oxygen, the degradation of phenolic compounds is initiated by microorganisms through carboxylation, fumarate addition to the methyl moiety or anoxic hydroxylation of the methyl moiety. Comparatively, under aerobic condition, the initiation mechanisms are revealed to be monoxygenation or dihydroxylation for phenol and ring hydroxylation or methyl group oxidation for cresols. While several studies biochemically characterized the enzymes and reaction mechanisms in the relevant degradation pathways, isotope fractionation patterns were rarely reported possibly due to constraints in current analytical methods. In this study, the carbon isotope fractionation patterns upon the degradation of phenol and cresols by several strains were analyzed by using isotope ratio mass spectrometry connected with liquid chromatography (LC-IRMS). The corresponding enrichment factors for carbon (ƐC) have been obtained. Cresols degradation by various strains showed generally moderate carbon isotope fractionation patterns with notable differences. For p-cresol degradation, five strains were examined. The aerobic strain Acinetobacter calcoaceticus NCIMB8250 exploits ring hydroxylation by molecular oxygen as initial reaction, and a ƐC value of -1.4±0.2‰ was obtained. Pseudomonas pseudoalcaligenes NCIMB 9867, an aerobic strain initiating cresols degradation via oxygen-dependent side chain hydroxylation, yielded a ƐC value of -2.3±0.2‰. Under nitrate-reducing conditions, Geobacter metallireducens DSM 7210 and Azoarcus buckelii DSM 14744 attacks p-cresol at the side chain by monohydroxylation using water as oxygen source; the two strains produced ƐC values of -3.6±0.4‰ and -2±0.1‰, accordingly. The sulfate-reducing Desulfosarcina cetonica DSM 7267 activating cresols by fumarate addition to the methyl moiety yielded ƐC values of -1.9±0.2‰ for p

  5. Subsurface clade of Geobacteraceae that predominates in a diversity of Fe(III)-reducing subsurface environments

    Science.gov (United States)

    Holmes, Dawn E.; O'Neil, Regina A.; Vrionis, Helen A.; N'Guessan, Lucie A.; Ortiz-Bernad, Irene; Larrahondo, Maria J.; Adams, Lorrie A.; Ward, Joy A.; Nicoll , Julie S.; Nevin, Kelly P.; Chavan, Milind A.; Johnson, Jessica P.; Long, Philip E.; Lovely, Derek R.

    2007-01-01

    There are distinct differences in the physiology of Geobacter species available in pure culture. Therefore, to understand the ecology of Geobacter species in subsurface environments, it is important to know which species predominate. Clone libraries were assembled with 16S rRNA genes and transcripts amplified from three subsurface environments in which Geobacter species are known to be important members of the microbial community: (1) a uranium-contaminated aquifer located in Rifle, CO, USA undergoing in situ bioremediation; (2) an acetate-impacted aquifer that serves as an analog for the long-term acetate amendments proposed for in situ uranium bioremediation and (3) a petroleum-contaminated aquifer in which Geobacter species play a role in the oxidation of aromatic hydrocarbons coupled with the reduction of Fe(III). The majority of Geobacteraceae 16S rRNA sequences found in these environments clustered in a phylogenetically coherent subsurface clade, which also contains a number of Geobacter species isolated from subsurface environments. Concatamers constructed with 43 Geobacter genes amplified from these sites also clustered within this subsurface clade. 16S rRNA transcript and gene sequences in the sediments and groundwater at the Rifle site were highly similar, suggesting that sampling groundwater via monitoring wells can recover the most active Geobacter species. These results suggest that further study of Geobacter species in the subsurface clade is necessary to accurately model the behavior of Geobacter species during subsurface bioremediation of metal and organic contaminants.

  6. ORF Alignment: NC_002939 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available Geobacter sulfurreducens PCA] ... Length = 81 ... Query: 1 ... MFVXXXXXXXXXXXXXXKGKRGIVKSILARARQDFNVSAAEVDL...QDVPDEAVLAFATVTG 60 ... MFV ... KGKRGIVKSILARARQDFNVSAAEVDLQDVPDEA...VLAFATVTG Sbjct: 1 ... MFVHSLCLHLHLPSHSLKGKRGIVKSILARARQDFNVSAAEVDLQDVPDEAVLAFATVTG 60 ...

  7. ORF Alignment: NC_002939 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available FliM ... [Geobacter sulfurreducens PCA] ... Length = 79 ... Query: 248 RLSKELLEAPLDVSVEVGGAVISLNDLLSLVPGDTIMLDTPCTSDLT...VKVGGVPKFTGMP 307 ... RLSKELLEAPLDVSVEVGGAVISLNDLLSLVPGDTIMLDTPCTSDLT...VKVGGVPKFTGMP Sbjct: 1 ... RLSKELLEAPLDVSVEVGGAVISLNDLLSLVPGDTIMLDTPCTSDLTVKVGGVPKFTGMP 60 ...

  8. ORF Alignment: NC_002939 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available GESSFNPLAINYNSNGTYDYGLMQINSSWEPALRK 79 ... ASAFCFEEAGNRYGISPQLLYAISKGESSFNPLAINYNSNGTYDYGLMQINSS...WEPALRK Sbjct: 1 ... ASAFCFEEAGNRYGISPQLLYAISKGESSFNPLAINYNSNGTYDYGLMQINSSWEPALRK 60 ... ...tein ... [Geobacter sulfurreducens PCA] ... Length = 101 ... Query: 20 ... ASAFCFEEAGNRYGISPQLLYAISK

  9. ORF Alignment: NC_002939 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available tein-alanine acetyltransferase [Geobacter ... sulfurreducens PCA] ... Length = 146 ... Query: 1 ... MNTADCP...TIMPMTEGDLDEVLQIESDSFPRPWTRDHFVAELASPRSFPVVARSPGGLIVG 60 ... MNTADCP...TIMPMTEGDLDEVLQIESDSFPRPWTRDHFVAELASPRSFPVVARSPGGLIVG Sbjct: 1 ... MNTADCPTIMPMTEGDLDEVLQIESDSFPRPWTRDHFVAEL

  10. NCBI nr-aa BLAST: CBRC-DNOV-01-1221 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-1221 ref|YP_001230970.1| UBA/THIF-type NAD/FAD binding protein [Geobacter uranium...reducens Rf4] gb|ABQ26397.1| UBA/THIF-type NAD/FAD binding protein [Geobacter uraniumreducens Rf4] YP_001230970.1 3.9 50% ...

  11. NCBI nr-aa BLAST: CBRC-BTAU-01-2822 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-2822 ref|YP_001230511.1| lipolytic enzyme, G-D-S-L family [Geobacter uranium...reducens Rf4] gb|ABQ25938.1| lipolytic enzyme, G-D-S-L family [Geobacter uraniumreducens Rf4] YP_001230511.1 0.001 40% ...

  12. NCBI nr-aa BLAST: CBRC-XTRO-01-0152 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-0152 ref|YP_001230889.1| inner-membrane translocator [Geobacter uranium...reducens Rf4] gb|ABQ26316.1| inner-membrane translocator [Geobacter uraniumreducens Rf4] YP_001230889.1 3e-34 36% ...

  13. NCBI nr-aa BLAST: CBRC-DMEL-04-0042 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available ike protein [Geobacter uraniumreducens Rf4] gb|ABQ26219.1| Uncharacterized membrane-associated protein-like protein [Geobacter uraniumreducens Rf4] YP_001230792.1 4.3 28% ... ...CBRC-DMEL-04-0042 ref|YP_001230792.1| Uncharacterized membrane-associated protein-l

  14. NCBI nr-aa BLAST: CBRC-DYAK-02-0022 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DYAK-02-0022 ref|YP_001232964.1| ATP synthase F0, A subunit [Geobacter uranium...reducens Rf4] gb|ABQ28391.1| ATP synthase F0, A subunit [Geobacter uraniumreducens Rf4] YP_001232964.1 0.85 25% ...

  15. NCBI nr-aa BLAST: CBRC-LAFR-01-0322 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0322 ref|YP_001231537.1| carbon starvation protein CstA [Geobacter uranium...reducens Rf4] gb|ABQ26964.1| carbon starvation protein CstA [Geobacter uraniumreducens Rf4] YP_001231537.1 0.96 31% ...

  16. Magnetite Compensates for the Lack of a Pilin-Associated c-Type Cytochrome in Extracellular Electron Exchange

    DEFF Research Database (Denmark)

    Liu, Fanghua; Rotaru, Amelia-Elena; Shrestha, Pravin

    2015-01-01

    investigation revealed that magnetite attached to the electrically conductive pili of Geobacter species in a manner reminiscent of the association of the multi-heme c-type cytochrome OmcS with the pili of Geobacter sulfurreducens. Magnetite conferred extracellular electron capabilities on an Omc...

  17. Carboxydotrophic growth of

    NARCIS (Netherlands)

    Geelhoed, J.; Henstra, A.M.; Stams, A.J.M.

    2016-01-01

    This study shows that Geobacter sulfurreducensgrows on carbon monoxide (CO) as electron donor with fumarateas electron acceptor. Geobacter sulfurreducens wastolerant to high CO levels, with up to 150 kPa in the headspacetested. During growth, hydrogen was detected in very slightamounts (~5 Pa). In

  18. NCBI nr-aa BLAST: CBRC-GACU-16-0048 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GACU-16-0048 ref|YP_001231116.1| lipopolysaccharide biosynthesis protein [Geobacter uranium...reducens Rf4] gb|ABQ26543.1| lipopolysaccharide biosynthesis protein [Geobacter uraniumreducens Rf4] YP_001231116.1 0.14 18% ...

  19. NCBI nr-aa BLAST: CBRC-HSAP-11-0402 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-HSAP-11-0402 ref|YP_001232090.1| heavy metal efflux pump, CzcA family [Geobacter uranium...reducens Rf4] gb|ABQ27517.1| heavy metal efflux pump, CzcA family [Geobacter uraniumreducens Rf4] YP_001232090.1 1.2 30% ...

  20. ORF Alignment: NC_002939 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002939 gi|39996821 >1v7zA 3 252 1 232 3e-42 ... ref|NP_952772.1| creatinine amidoh...ydrolase [Geobacter sulfurreducens PCA] ... gb|AAR35099.1| creatinine amidohydrolase [Geobacter ...

  1. An Investigation of Civilians Preparedness to Compete with Individuals with Military Experience for Army Board Select Acquisition Positions

    Science.gov (United States)

    2017-05-25

    supervisor’s name and correct email address are 12 listed in your IDP. If your current supervisor is not listed in your IDP, please advise your...and General Staff College (LTC/GS-14 level boards) and/or Senior Staff College (COL/GS-15 level boards). Potential board members are nominated by

  2. Set anode potentials affect the electron fluxes and microbial community structure in propionate-fed microbial electrolysis cells

    KAUST Repository

    Rao, Hari Ananda; Katuri, Krishna; Logan, Bruce E.; Saikaly, Pascal

    2016-01-01

    , but their relative abundance varied among the tested SAPs. Microbial community analysis implies that complete degradation of propionate in all the tested SAPs was facilitated by syntrophic interactions between fermenters and Geobacter at the anode and ferementers

  3. Functional environmental proteomics: elucidating the role of a c-type cytochrome abundant during uranium bioremediation.

    Science.gov (United States)

    Yun, Jiae; Malvankar, Nikhil S; Ueki, Toshiyuki; Lovley, Derek R

    2016-02-01

    Studies with pure cultures of dissimilatory metal-reducing microorganisms have demonstrated that outer-surface c-type cytochromes are important electron transfer agents for the reduction of metals, but previous environmental proteomic studies have typically not recovered cytochrome sequences from subsurface environments in which metal reduction is important. Gel-separation, heme-staining and mass spectrometry of proteins in groundwater from in situ uranium bioremediation experiments identified a putative c-type cytochrome, designated Geobacter subsurface c-type cytochrome A (GscA), encoded within the genome of strain M18, a Geobacter isolate previously recovered from the site. Homologs of GscA were identified in the genomes of other Geobacter isolates in the phylogenetic cluster known as subsurface clade 1, which predominates in a diversity of Fe(III)-reducing subsurface environments. Most of the gscA sequences recovered from groundwater genomic DNA clustered in a tight phylogenetic group closely related to strain M18. GscA was most abundant in groundwater samples in which Geobacter sp. predominated. Expression of gscA in a strain of Geobacter sulfurreducens that lacked the gene for the c-type cytochrome OmcS, thought to facilitate electron transfer from conductive pili to Fe(III) oxide, restored the capacity for Fe(III) oxide reduction. Atomic force microscopy provided evidence that GscA was associated with the pili. These results demonstrate that a c-type cytochrome with an apparent function similar to that of OmcS is abundant when Geobacter sp. are abundant in the subsurface, providing insight into the mechanisms for the growth of subsurface Geobacter sp. on Fe(III) oxide and suggesting an approach for functional analysis of other Geobacter proteins found in the subsurface.

  4. Novel mode of microbial energy metabolism: organic carbon oxidation coupled to dissimilatory reduction of iron or manganese.

    Science.gov (United States)

    Lovley, D R; Phillips, E J

    1988-06-01

    A dissimilatory Fe(III)- and Mn(IV)-reducing microorganism was isolated from freshwater sediments of the Potomac River, Maryland. The isolate, designated GS-15, grew in defined anaerobic medium with acetate as the sole electron donor and Fe(III), Mn(IV), or nitrate as the sole electron acceptor. GS-15 oxidized acetate to carbon dioxide with the concomitant reduction of amorphic Fe(III) oxide to magnetite (Fe(3)O(4)). When Fe(III) citrate replaced amorphic Fe(III) oxide as the electron acceptor, GS-15 grew faster and reduced all of the added Fe(III) to Fe(II). GS-15 reduced a natural amorphic Fe(III) oxide but did not significantly reduce highly crystalline Fe(III) forms. Fe(III) was reduced optimally at pH 6.7 to 7 and at 30 to 35 degrees C. Ethanol, butyrate, and propionate could also serve as electron donors for Fe(III) reduction. A variety of other organic compounds and hydrogen could not. MnO(2) was completely reduced to Mn(II), which precipitated as rhodochrosite (MnCO(3)). Nitrate was reduced to ammonia. Oxygen could not serve as an electron acceptor, and it inhibited growth with the other electron acceptors. This is the first demonstration that microorganisms can completely oxidize organic compounds with Fe(III) or Mn(IV) as the sole electron acceptor and that oxidation of organic matter coupled to dissimilatory Fe(III) or Mn(IV) reduction can yield energy for microbial growth. GS-15 provides a model for how enzymatically catalyzed reactions can be quantitatively significant mechanisms for the reduction of iron and manganese in anaerobic environments.

  5. Development of a Systems Engineering Competency Model Tool for the Aviation and Missile Research, Development, And Engineering Center (AMRDEC)

    Science.gov (United States)

    2017-06-01

    grade level (GS-7 to GS-15). This foundational model is structured to support the individual needs of any Department of Defense organization and is... organizational level with traceability to the approved OPM competencies. The Redstone SECCM Tool will allow documentation of system engineering competencies and...assessment of individual and organizational development and training needs. This report documents the requirements analysis, system design, and system

  6. Department of Clinical Investigation Annual Research Progress Report Fiscal Year 2002

    Science.gov (United States)

    2002-09-30

    on ice for 20 min. Samples will be centrifuged at 4ºC for 20 min. at 9,500 rpm in a Beckman GS-15R tabletop centrifuge. The aqueous layer will be...positive serology by the presence of either positive H. pylori on histology and a positive rapid urease test. At endoscopy, after aspiration of gastric...laboratory tests and Puality of Life questionnare completed; digital photography will be performed; bi- layer tracing of the wound will be measured

  7. Combining microbial cultures for efficient production of electricity from butyrate in a microbial electrochemical cell

    Science.gov (United States)

    Miceli, Joseph F.; Garcia-Peña, Ines; Parameswaran, Prathap; Torres, César I.; Krajmalnik-Brown, Rosa

    2014-01-01

    Butyrate is an important product of anaerobic fermentation; however, it is not directly used by characterized strains of the highly efficient anode respiring bacteria (ARB) Geobacter sulfurreducens in microbial electrochemical cells. By combining a butyrate-oxidizing community with a Geobacter rich culture, we generated a microbial community which outperformed many naturally derived communities found in the literature for current production from butyrate and rivaled the highest performing natural cultures in terms of current density (~11 A/m2) and Coulombic efficiency (~70%). Microbial community analyses support the shift in the microbial community from one lacking efficient ARB in the marine hydrothermal vent community to a community consisting of ~80% Geobacter in the anode biofilm. This demonstrates the successful production and adaptation of a novel microbial culture for generating electrical current from butyrate with high current density and high Coulombic efficiency, by combining two mixed micro bial cultures containing complementing biochemical pathways. PMID:25048958

  8. Potential for Methanosarcina to contribute to uranium reduction during acetate-promoted groundwater bioremediation

    DEFF Research Database (Denmark)

    Holmes, Dawn E; Orellana, Roberto; Giloteaux, Ludovic

    2018-01-01

    Previous studies of acetate-promoted bioremediation of uranium-contaminated aquifers focused on Geobacter because no other microorganisms that can couple the oxidation of acetate with U(VI) reduction had been detected in situ. Monitoring the levels of methyl CoM reductase subunit A (mcrA) transcr......Previous studies of acetate-promoted bioremediation of uranium-contaminated aquifers focused on Geobacter because no other microorganisms that can couple the oxidation of acetate with U(VI) reduction had been detected in situ. Monitoring the levels of methyl CoM reductase subunit A (mcr......(VI) reduction was observed in inactive controls. These results demonstrate that Methanosarcina species could play an important role in the long-term bioremediation of uranium-contaminated aquifers after depletion of Fe(III) oxides limits the growth of Geobacter species. The results also suggest...

  9. Influence of heterogeneous ammonium availability on bacterial community structure and the expression of nitrogen fixation and ammonium transporter genes during in situ bioremediation of uranium-contaminated groundwater

    Energy Technology Data Exchange (ETDEWEB)

    Mouser, P.J.; N' Guessan, A.L.; Elifantz, H.; Holmes, D.E.; Williams, K.H.; Wilkins, M.J.; Long, P.E.; Lovley, D.R.

    2009-04-01

    The impact of ammonium availability on microbial community structure and the physiological status and activity of Geobacter species during in situ bioremediation of uranium-contaminated groundwater was evaluated. Ammonium concentrations varied by as much as two orders of magnitude (<4 to 400 {micro}M) across the study site. Analysis of 16S rRNA gene sequences suggested that ammonium influenced the composition of the microbial community prior to acetate addition with Rhodoferax species predominating over Geobacter species at the site with the highest ammonium, and Dechloromonas species dominating at sites with lowest ammonium. However, once acetate was added, and dissimilatory metal reduction was stimulated, Geobacter species became the predominant organisms at all locations. Rates of U(VI) reduction appeared to be more related to the concentration of acetate that was delivered to each location rather than the amount of ammonium available in the groundwater. In situ mRNA transcript abundance of the nitrogen fixation gene, nifD, and the ammonium importer gene, amtB, in Geobacter species indicated that ammonium was the primary source of nitrogen during in situ uranium reduction, and that the abundance of amtB transcripts was inversely correlated to ammonium levels across all sites examined. These results suggest that nifD and amtB expression by subsurface Geobacter species are closely regulated in response to ammonium availability to ensure an adequate supply of nitrogen while conserving cell resources. Thus, quantifying nifD and amtB expression appears to be a useful approach for monitoring the nitrogen-related physiological status of Geobacter species in subsurface environments during bioremediation. This study also emphasizes the need for more detailed analysis of geochemical/physiological interactions at the field scale, in order to adequately model subsurface microbial processes.

  10. Microbial Fuel Cells; Microbiele Fuel Cells

    Energy Technology Data Exchange (ETDEWEB)

    Louzada, K.

    2012-03-15

    Geobacter bacteria are able to transfer electrons to their environment and have a recovering effect on the environment. The US Office for Naval research (ONR) is conducting research on the application of bacteria in the development of natural batteries for under water applications such as lighting, chemical sensors, hydrophones and autonomous vehicles. [Dutch] Geobacter bacterien zijn in staat zijn om elektronen over te dragen aan hun omgeving en hebben een herstellende invloed op het milieu. Het US Office for Naval Research (ONR) voert onderzoek uit naar de toepassingen van de bacterien in de ontwikkeling van natuurlijke batterijen voor onderwater applicaties als verlichting, chemische sensoren, hydrofonen en autonome voertuigen.

  11. Potential for Methanosarcina to contribute to uranium reduction during acetate-promoted groundwater bioremediation

    DEFF Research Database (Denmark)

    Holmes, Dawn E; Orellana, Roberto; Giloteaux, Ludovic

    2017-01-01

    Previous studies of in situ bioremediation of uranium-contaminated groundwater with acetate injections have focused on the role of Geobacter species in U(VI) reduction because of a lack of other abundant known U(VI)-reducing microorganisms. Monitoring the levels of methyl CoM reductase subunit...... an important role in the long-term bioremediation of uranium-contaminated aquifers after depletion of Fe(III) oxides limits the growth of Geobacter species. The results also suggest that Methanosarcina have the potential to influence uranium geochemistry in a diversity of anaerobic sedimentary environments....

  12. Microbial Activity Influences Electrical Conductivity of Biofilm Anode

    Science.gov (United States)

    This study assessed the conductivity of a Geobacter-enriched biofilm anode along with biofilm activity in a microbial electrochemical cell (MxC) equipped with two gold anodes (25 mM acetate medium), as different proton gradients were built throughout the biofilm. There was no pH ...

  13. Quantification of the methane concentration using anaerobic oxidation of methane coupled to extracellular electron transfer

    Science.gov (United States)

    A biofilm anode acclimated with acetate, acetate+methane, and methane growth media for over three years produced a steady current density of 1.6-2.3 mA/m^2 in a microbial electrochemical cell (MxC) fed with methane as the sole electron donor. Geobacter was the dominant genus for...

  14. Influence of ammonium availability on expression of nifD and amtB genes during biostimulation of a U(VI) contaminated aquifer: implications for U(VI) removal and monitoring the metabolic state of Geobacteraceae

    Energy Technology Data Exchange (ETDEWEB)

    Mouser, Paula J.; N' Guessan, A. Lucie; Elifantz, Hila; Holmes, Dawn E.; Williams, Kenneth H; Wilkins, Michael J.; Long, Philip E.; Lovley, Derek R.

    2009-03-25

    The influence of ammonium availability on bacterial community structure and the physiological status of Geobacter species during in situ bioremediation of uranium-contaminated groundwater was evaluated. Ammonium concentrations varied by 2 orders of magnitude (<4 to 400 ?M) across the study site. Analysis of 16S rRNA sequences suggested that ammonium may have been one factor influencing the community composition prior to acetate amendment with Rhodoferax species predominating over Geobacter species with higher ammonium and Dechloromonas species dominating at the site with lowest ammonium. However, once acetate was added and dissimilatory metal reduction was stimulated, Geobacter species became the predominant organisms at all locations. Rates of U(VI) reduction appeared to be more related to acetate concentrations rather than ammonium levels. In situ mRNA transcript abundance of the nitrogen fixation gene, nifD, and the ammonium transporter gene, amtB, in Geobacter species indicated that ammonium was the primary source of nitrogen during uranium reduction. The abundance of amtB was inversely correlated to ammonium levels, whereas nifD transcript levels were similar across all sites examined. These results suggest that nifD and amtB expression are closely regulated in response to ammonium availability to ensure an adequate supply of nitrogen while conserving cell resources. Thus, quantifying nifD and amtB transcript expression appears to be a useful approach for monitoring the nitrogen-related physiological status of subsurface Geobacter species. This study also emphasizes the need for more detailed analysis of geochemical and physiological interactions at the field scale in order to adequately model subsurface microbial processes during bioremediation.

  15. Correlation between microbial community and granule conductivity in anaerobic bioreactors for brewery wastewater treatment

    DEFF Research Database (Denmark)

    Shrestha, Pravin; Malvankar, Nikhil S.; Werner, Jeffrey

    2014-01-01

    Prior investigation of an upflow anaerobic sludge blanket (UASB) reactor treating brewery wastes suggested that direct interspecies electron transfer (DIET) significantly contributed to interspecies electron transfer to methanogens. To investigate DIET in granules further, the electrical conducti......Prior investigation of an upflow anaerobic sludge blanket (UASB) reactor treating brewery wastes suggested that direct interspecies electron transfer (DIET) significantly contributed to interspecies electron transfer to methanogens. To investigate DIET in granules further, the electrical...... conductivity and bacterial community composition of granules in fourteen samples from four different UASB reactors treating brewery wastes were investigated. All of the UASB granules were electrically conductive whereas control granules from ANAMMOX (ANaerobic AMMonium OXidation) reactors and microbial...... granules from an aerobic bioreactor designed for phosphate removal were not. There was a moderate correlation (r = 0.67) between the abundance of Geobacter species in the UASB granules and granule conductivity, suggesting that Geobacter contributed to granule conductivity. These results, coupled...

  16. Transcriptomic and genetic analysis of direct interspecies electron transfer

    DEFF Research Database (Denmark)

    Shrestha, Pravin Malla; Rotaru, Amelia-Elena; Summers, Zarath M

    2013-01-01

    The possibility that metatranscriptomic analysis could distinguish between direct interspecies electron transfer (DIET) and H2 interspecies transfer (HIT) in anaerobic communities was investigated by comparing gene transcript abundance in cocultures in which Geobacter sulfurreducens....... These results demonstrate that there are unique gene expression patterns that distinguish DIET from HIT and suggest that metatranscriptomics may be a promising route to investigate interspecies electron transfer pathways in more-complex environments....

  17. Rust dissolution and removal by iron-reducing bacteria: A potential rehabilitation of rusted equipment

    International Nuclear Information System (INIS)

    Starosvetsky, J.; Kamari, R.; Farber, Y.; Bilanović, D.; Armon, R.

    2016-01-01

    Highlights: • The present study demonstrated the high reductive capacity of both strains: the collection S. oneidensis and the wild strain Geobacter spp. (soil isolate). • The experimental strains were successful in Fe 3+ reduction for both states: soluble and crystalline (originally prepared from rust). • Rust dissolution can be improved by: addition of AFC at low concentration (0.2 g/l), increasing bacterial initial inoculum and rust reactive surface. • Both experimental IRB strains were able to completely remove previously formed rust on carbon steel coupons. • Additional results (not showed) revealed that culture S. oneidensis and the environmental isolate Geobacter spp., apparently have a different mechanism of iron reduction that requires further study. - Abstract: Iron reducing bacteria (IRB), to be used in rust dissolution and removal, have been isolated and enriched from different environmental sources. Comparative measurements revealed that a soil isolate (Geobacter sulfurreducens sp.) had the highest reductive activity equivalent to Shewanella oneidensis (strain CIP 106686, pure culture). Both reductive microorganisms can use Fe 3+ ions as electron acceptors from soluble as well as from crystalline sources. In nutrient medium containing soluble Fe 3+ , the highest reductive activity obtained for G. sulfurreducens sp. and S. oneidensis was 93 and 97% respectively. Successful removal of rust from carbon steel coupons has been achieved with both experimental bacteria.

  18. Pulse electromagnetic fields enhance extracellular electron transfer in magnetic bioelectrochemical systems.

    Science.gov (United States)

    Zhou, Huihui; Liu, Bingfeng; Wang, Qisong; Sun, Jianmin; Xie, Guojun; Ren, Nanqi; Ren, Zhiyong Jason; Xing, Defeng

    2017-01-01

    Microbial extracellular electron transfer (EET) is essential in driving the microbial interspecies interaction and redox reactions in bioelectrochemical systems (BESs). Magnetite (Fe 3 O 4 ) and magnetic fields (MFs) were recently reported to promote microbial EET, but the mechanisms of MFs stimulation of EET and current generation in BESs are not known. This study investigates the behavior of current generation and EET in a state-of-the-art pulse electromagnetic field (PEMF)-assisted magnetic BES (PEMF-MBES), which was equipped with magnetic carbon particle (Fe 3 O 4 @N-mC)-coated electrodes. Illumina Miseq sequencing of 16S rRNA gene amplicons was also conducted to reveal the changes of microbial communities and interactions on the anode in response to magnetic field. PEMF had significant influences on current generation. When reactors were operated in microbial fuel cell (MFC) mode with pulse electromagnetic field (PEMF-MMFCs), power densities increased by 25.3-36.0% compared with no PEMF control MFCs (PEMF-OFF-MMFCs). More interestingly, when PEMF was removed, the power density dropped by 25.7%, while when PEMF was reintroduced, the value was restored to the previous level. Illumina sequencing of 16S rRNA gene amplicon and principal component analysis (PCA) based on operational taxonomic units (OTUs) indicate that PEMFs led to the shifts in microbial community and changes in species evenness that decreased biofilm microbial diversity. Geobacter spp. were found dominant in all anode biofilms, but the relative abundance in PEMF-MMFCs (86.1-90.0%) was higher than in PEMF-OFF-MMFCs (82.5-82.7%), indicating that the magnetic field enriched Geobacter on the anode. The current generation of Geobacter -inoculated microbial electrolysis cells (MECs) presented the same change regularity, the accordingly increase or decrease corresponding with switch of PEMF, which confirmed the reversible stimulation of PEMFs on microbial electron transfer. The pulse electromagnetic

  19. Analog Design for Digital Deployment of a Serious Leadership Game

    Science.gov (United States)

    Maxwell, Nicholas; Lang, Tristan; Herman, Jeffrey L.; Phares, Richard

    2012-01-01

    This paper presents the design, development, and user testing of a leadership development simulation. The authors share lessons learned from using a design process for a board game to allow for quick and inexpensive revision cycles during the development of a serious leadership development game. The goal of this leadership simulation is to accelerate the development of leadership capacity in high-potential mid-level managers (GS-15 level) in a federal government agency. Simulation design included a mixed-method needs analysis, using both quantitative and qualitative approaches to determine organizational leadership needs. Eight design iterations were conducted, including three user testing phases. Three re-design iterations followed initial development, enabling game testing as part of comprehensive instructional events. Subsequent design, development and testing processes targeted digital application to a computer- and tablet-based environment. Recommendations include pros and cons of development and learner testing of an initial analog simulation prior to full digital simulation development.

  20. Gene expression correlates with process rates quantified for sulfate- and Fe(III-reducing bacteria in U(VI-contaminated sediments

    Directory of Open Access Journals (Sweden)

    Denise M Akob

    2012-08-01

    Full Text Available Though iron- and sulfate-reducing bacteria are well known for mediating uranium(VI reduction in contaminated subsurface environments, quantifying the in situ activity of the microbial groups responsible remains a challenge. The objective of this study was to demonstrate the use of quantitative molecular tools that target mRNA transcripts of key genes related to Fe(III and sulfate reduction pathways in order to monitor these processes during in situ U(VI remediation in the subsurface. Expression of the Geobacteraceae-specific citrate synthase gene (gltA and the dissimilatory (bisulfite reductase gene (dsrA, were correlated with the activity of iron- or sulfate-reducing microorganisms, respectively, under stimulated bioremediation conditions in microcosms of sediments sampled from the U.S. Department of Energy’s Oak Ridge Integrated Field Research Challenge (OR-IFRC site at Oak Ridge, Tennessee. In addition, Geobacteraceae-specific gltA and dsrA transcript levels were determined in parallel with the predominant electron acceptors present in moderately and highly contaminated subsurface sediments from the OR-IFRC. Phylogenetic analysis of the cDNA generated from dsrA mRNA, sulfate-reducing bacteria-specific 16S rRNA, and gltA mRNA identified activity of specific microbial groups. Active sulfate reducers were members of the Desulfovibrio, Desulfobacterium, and Desulfotomaculum genera. Members of the subsurface Geobacter clade, closely related to uranium-reducing Geobacter uraniireducens and Geobacter daltonii, were the metabolically-active iron-reducers in biostimulated microcosms and in situ core samples. Direct correlation of transcripts and process rates demonstrated evidence of competition between the functional guilds in subsurface sediments. We further showed that active populations of Fe(III-reducing bacteria and sulfate-reducing bacteria are present in OR-IFRC sediments and are good potential targets for in situ bioremediation.

  1. Convergent development of anodic bacterial communities in microbial fuel cells.

    KAUST Repository

    Yates, Matthew D

    2012-05-10

    Microbial fuel cells (MFCs) are often inoculated from a single wastewater source. The extent that the inoculum affects community development or power production is unknown. The stable anodic microbial communities in MFCs were examined using three inocula: a wastewater treatment plant sample known to produce consistent power densities, a second wastewater treatment plant sample, and an anaerobic bog sediment. The bog-inoculated MFCs initially produced higher power densities than the wastewater-inoculated MFCs, but after 20 cycles all MFCs on average converged to similar voltages (470±20 mV) and maximum power densities (590±170 mW m(-2)). The power output from replicate bog-inoculated MFCs was not significantly different, but one wastewater-inoculated MFC (UAJA3 (UAJA, University Area Joint Authority Wastewater Treatment Plant)) produced substantially less power. Denaturing gradient gel electrophoresis profiling showed a stable exoelectrogenic biofilm community in all samples after 11 cycles. After 16 cycles the predominance of Geobacter spp. in anode communities was identified using 16S rRNA gene clone libraries (58±10%), fluorescent in-situ hybridization (FISH) (63±6%) and pyrosequencing (81±4%). While the clone library analysis for the underperforming UAJA3 had a significantly lower percentage of Geobacter spp. sequences (36%), suggesting that a predominance of this microbe was needed for convergent power densities, the lower percentage of this species was not verified by FISH or pyrosequencing analyses. These results show that the predominance of Geobacter spp. in acetate-fed systems was consistent with good MFC performance and independent of the inoculum source.

  2. Thermodynamic analysis of regulation in metabolic networks using constraint-based modeling

    Directory of Open Access Journals (Sweden)

    Mahadevan Radhakrishnan

    2010-05-01

    Full Text Available Abstract Background Geobacter sulfurreducens is a member of the Geobacter species, which are capable of oxidation of organic waste coupled to the reduction of heavy metals and electrode with applications in bioremediation and bioenergy generation. While the metabolism of this organism has been studied through the development of a stoichiometry based genome-scale metabolic model, the associated regulatory network has not yet been well studied. In this manuscript, we report on the implementation of a thermodynamics based metabolic flux model for Geobacter sulfurreducens. We use this updated model to identify reactions that are subject to regulatory control in the metabolic network of G. sulfurreducens using thermodynamic variability analysis. Findings As a first step, we have validated the regulatory sites and bottleneck reactions predicted by the thermodynamic flux analysis in E. coli by evaluating the expression ranges of the corresponding genes. We then identified ten reactions in the metabolic network of G. sulfurreducens that are predicted to be candidates for regulation. We then compared the free energy ranges for these reactions with the corresponding gene expression fold changes under conditions of different environmental and genetic perturbations and show that the model predictions of regulation are consistent with data. In addition, we also identify reactions that operate close to equilibrium and show that the experimentally determined exchange coefficient (a measure of reversibility is significant for these reactions. Conclusions Application of the thermodynamic constraints resulted in identification of potential bottleneck reactions not only from the central metabolism but also from the nucleotide and amino acid subsystems, thereby showing the highly coupled nature of the thermodynamic constraints. In addition, thermodynamic variability analysis serves as a valuable tool in estimating the ranges of ΔrG' of every reaction in the model

  3. Enrichment of specific protozoan populations during in situ bioremediation of uranium-contaminated groundwater

    Energy Technology Data Exchange (ETDEWEB)

    Holmes, Dawn; Giloteaux, L.; Williams, Kenneth H.; Wrighton, Kelly C.; Wilkins, Michael J.; Thompson, Courtney A.; Roper, Thomas J.; Long, Philip E.; Lovley, Derek

    2013-07-28

    The importance of bacteria in the anaerobic bioremediation of groundwater polluted with organic and/or metal contaminants is well-recognized and in some instances so well understood that modeling of the in situ metabolic activity of the relevant subsurface microorganisms in response to changes in subsurface geochemistry is feasible. However, a potentially significant factor influencing bacterial growth and activity in the subsurface that has not been adequately addressed is protozoan predation of the microorganisms responsible for bioremediation. In field experiments at a uranium-contaminated aquifer located in Rifle, CO, acetate amendments initially promoted the growth of metal-reducing Geobacter species followed by the growth of sulfate-reducers, as previously observed. Analysis of 18S rRNA gene sequences revealed a broad diversity of sequences closely related to known bacteriovorous protozoa in the groundwater prior to the addition of acetate. The bloom of Geobacter species was accompanied by a specific enrichment of sequences most closely related to the amoeboid flagellate, Breviata anathema, which at their peak accounted for over 80% of the sequences recovered. The abundance of Geobacter species declined following the rapid emergence of B. anathema. The subsequent growth of sulfate-reducing Peptococcaceae was accompanied by another specific enrichment of protozoa, but with sequences most similar to diplomonadid flagellates from the family Hexamitidae, which accounted for up to 100% of the sequences recovered during this phase of the bioremediation. These results suggest a prey-predator response with specific protozoa responding to increased availability of preferred prey bacteria. Thus, quantifying the influence of protozoan predation on the growth, activity, and composition of the subsurface bacterial community is essential for predictive modeling of in situ uranium bioremediation strategies.

  4. Molecular analysis of phosphate limitation in Geobacteraceae during the bioremediation of a uranium-contaminated aquifer

    Energy Technology Data Exchange (ETDEWEB)

    N' Guessan, L.A.; Elifantz, H.; Nevin, K.P.; Mouser, P.J.; Methe, B.; Woodard, T. L.; Manley, K.; Williams, K. H.; Wilkins, M. J.; Larsen, J.T.; Long, P. E.; Lovley, D. R.

    2009-09-01

    Nutrient limitation is an environmental stress that may reduce the effectiveness of bioremediation strategies, especially when the contaminants are organic compounds or when organic compounds are added to promote microbial activities such as metal reduction. Genes indicative of phosphate-limitation were identified via microarray analysis of chemostat cultures of Geobacter sulfureducens. This analysis revealed that genes in the pst-pho operon, which is associated with a high affinity phosphate uptake system in other microorganisms, had significantly higher transcript abundance under phosphate-limiting conditions, with the genes pstB and phoU the most up-regulated. Quantitative PCR analysis of pstB and phoU transcript levels in G. sulfurreducens grown in chemostats demonstrated that the expression of these genes increased when phosphate was removed from the culture medium. Transcripts of pstB and phoU within the subsurface Geobacter species predominating during an in situ uranium bioremediation field experiment were more abundant than in chemostat cultures of G. sulfurreducens that were not limited for phosphate. Addition of phosphate to incubations of subsurface sediments did not stimulate dissimilatory metal reduction. The added phosphate was rapidly adsorbed onto the sediments. The results demonstrate that Geobacter species can effectively reduce U(VI) even when experiencing suboptimal phosphate concentrations and that increasing phosphate availability with phosphate additions is difficult to achieve due to the high reactivity of this compound. This transcript-based approach developed for diagnosing phosphate limitation should be applicable to assessing the potential need for additional phosphate in other bioremediation processes.

  5. Dicty_cDB: Contig-U12922-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 2e-11 2 ( EA053492 ) Sequence 221 from patent US 7157621. 62 2e-11 4 ( DY329960 ) OB_MEa0004O04.r OB_MEa Ocimum basil...hql*mvfiknfkkkkkkkk Frame B: llaywxyyyyskyifvyt*niyk*fllvvnfv*lisivsnf*nnkekkngxk...CP001124 |pid:none) Geobacter bemidjiensis Bem, comp... 70 2e-10 (A4JHJ8) RecName...... 56 4e-06 CP000852_1193( CP000852 |pid:none) Caldivirga maquilingensis IC-16... 55 5e-06 ...cc*frfw**awklw*i fr*sk*rgys*icmhfnfignqks*f*tksiinwwwyc*fy*cgchfqryh*sn*rvs** yh*iksfhfr*krwsklsigfttyernws*ity

  6. The electric picnic: synergistic requirements for exoelectrogenic microbial communities

    KAUST Repository

    Kiely, Patrick D

    2011-06-01

    Characterization of the various microbial populations present in exoelectrogenic biofilms provides insight into the processes required to convert complex organic matter in wastewater streams into electrical current in bioelectrochemical systems (BESs). Analysis of the community profiles of exoelectrogenic microbial consortia in BESs fed different substrates gives a clearer picture of the different microbial populations present in these exoelectrogenic biofilms. Rapid utilization of fermentation end products by exoelectrogens (typically Geobacter species) relieves feedback inhibition for the fermentative consortia, allowing for rapid metabolism of organics. Identification of specific syntrophic processes and the communities characteristic of these anodic biofilms will be a valuable aid in improving the performance of BESs. © 2011 Elsevier Ltd.

  7. Dicty_cDB: VFA628 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 8 |pid:none) Methanobrevibacter smithii ATCC ... 42 0.017 CP001390_1265( CP001390 |pid:none) Geobacter sp. F...000678_1111( CP000678 |pid:none) Methanobrevibacter smithii ATCC... 36 0.91 (Q9Z3...otrophomonas maltophilia K27... 36 1.2 CP000678_996( CP000678 |pid:none) Methanobrevibacter smithii ATCC ......:none) Methanobrevibacter smithii ATCC ... 34 3.5 protein update 2009. 6.16 PSORT psg: 0.85 gvh: 0.47 alm: 0

  8. Nuclear technology in research and everyday life; Kerntechnik in Forschung und Alltag

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2015-12-15

    The paper.. discusses the impact of nuclear technology in research and everyday life covering the following issues: miniaturization of memory devices, neutron radiography in material science, nuclear reactions in the universe, sterilization of food, medical applies, cosmetics and packaging materials using beta and gamma radiation, neutron imaging for radioactive waste analysis, microbial transformation of uranium (geobacter uraniireducens), nuclear technology knowledge preservation, spacecrafts voyager 1 and 2, future fusion power plants, prompt gamma activation analysis in archeology, radiation protection and radioecology and nuclear medicine (radiotherapy).

  9. Nuclear technology in research and everyday life

    International Nuclear Information System (INIS)

    2015-12-01

    The paper.. discusses the impact of nuclear technology in research and everyday life covering the following issues: miniaturization of memory devices, neutron radiography in material science, nuclear reactions in the universe, sterilization of food, medical applies, cosmetics and packaging materials using beta and gamma radiation, neutron imaging for radioactive waste analysis, microbial transformation of uranium (geobacter uraniireducens), nuclear technology knowledge preservation, spacecrafts voyager 1 and 2, future fusion power plants, prompt gamma activation analysis in archeology, radiation protection and radioecology and nuclear medicine (radiotherapy).

  10. Acetate availability and its influence on sustainable bioremediation of uranium-contaminated groundwater

    International Nuclear Information System (INIS)

    Williams, Kenneth H.; Long, Philip E.; Davis, James A.; Wilkins, Michael J.; N'Guessan, A. Lucie; Steefel, Carl I.; Yang, Li; Newcomer, Darrell R.; Spane, Frank A.; Kerkhof, L.; McGuinness, L.; Dayvault, Richard; Lovley, Derek

    2011-01-01

    Field biostimulation experiments at the U.S. Department of Energy's Integrated Field Research Challenge (IFRC) site in Rifle, Colorado, have demonstrated that uranium concentrations in groundwater can be decreased to levels below the U.S. Environmental Protection Agency's (EPA) drinking water standard (0.126 (micro)M). During successive summer experiments - referred to as 'Winchester' (2007) and 'Big Rusty' (2008) - acetate was added to the aquifer to stimulate the activity of indigenous dissimilatory metal-reducing bacteria capable of reductively immobilizing uranium. The two experiments differed in the length of injection (31 vs. 110 days), the maximum concentration of acetate (5 vs. 30 mM), and the extent to which iron reduction ('Winchester') or sulfate reduction ('Big Rusty') was the predominant metabolic process. In both cases, rapid removal of U(VI) from groundwater occurred at calcium concentrations (6 mM) and carbonate alkalinities (8 meq/L) where Ca-UO2-CO3 ternary complexes constitute >90% of uranyl species in groundwater. Complete consumption of acetate and increased alkalinity (>30 meq/L) accompanying the onset of sulfate reduction corresponded to temporary increases in U(VI); however, by increasing acetate concentrations in excess of available sulfate (10 mM), low U(VI) concentrations (0.1-0.05 (micro)M) were achieved for extended periods of time (>140 days). Uniform delivery of acetate during 'Big Rusty' was impeded due to decreases in injection well permeability, likely resulting from biomass accumulation and carbonate and sulfide mineral precipitation. Such decreases were not observed during the short-duration 'Winchester' experiment. Terminal restriction fragment length polymorphism (TRFLP) analysis of 16S rRNA genes demonstrated that Geobacter sp. and Geobacter-like strains dominated the groundwater community profile during iron reduction, with 13C stable isotope probing (SIP) results confirming these strains were actively utilizing acetate to

  11. Acetate availability and its influence on sustainable bioremediation of Uranium-contaminated groundwater

    Science.gov (United States)

    Williams, K.H.; Long, P.E.; Davis, J.A.; Wilkins, M.J.; N'Guessan, A. L.; Steefel, Carl; Yang, L.; Newcomer, D.; Spane, F.A.; Kerkhof, L.J.; Mcguinness, L.; Dayvault, R.; Lovley, D.R.

    2011-01-01

    Field biostimulation experiments at the U.S. Department of Energy's Integrated Field Research Challenge (IFRC) site in Rifle, Colorado, have demonstrated that uranium concentrations in groundwater can be decreased to levels below the U.S. Environmental Protection Agency's (EPA) drinking water standard (0.126??M).During successive summer experiments - referred to as "Winchester" (2007) and "Big Rusty" (2008) - acetate was added to the aquifer to stimulate the activity of indigenous dissimilatory metal reducing bacteria capable of reductively immobilizing uranium. The two experiments differed in the length of injection (31 vs. 110 days), the maximum concentration of acetate (5 vs. 30 mM),and the extent to which iron reduction ("Winchester") or sulfate reduction("Big Rusty") was the predominant metabolic process. In both cases, rapid removal of U(VI) from groundwater occurred at calcium concentrations (6 mM) and carbonate alkalinities (8 meq/L) where Ca-UO2-CO3 ternary complexes constitute >90% of uranyl species in groundwater. Complete consumption of acetate and increased alkalinity (>30 meq/L) accompanying the onset of sulfate reduction corresponded to temporary increases in U(VI);however, by increasing acetate concentrations in excess of available sulfate (10 mM), low U(VI) concentrations (0.1-0.05 ??M) were achieved for extended periods of time (>140 days). Uniform delivery of acetate during "Big Rusty" was impeded due to decreases in injection well permeability, likely resulting from biomass accumulation and carbonate and sulfide mineral precipitation. Such decreases were not observed during the short-duration "Winchester" experiment. Terminal restriction fragment length polymorphism (TRFLP) analysis of 16S rRNA genes demonstrated that Geobacter sp. and Geobacter-like strains dominated the groundwater community profile during iron reduction, with 13C stable isotope probing (SIP) results confirming these strains were actively utilizing acetate to replicate their

  12. Set anode potentials affect the electron fluxes and microbial community structure in propionate-fed microbial electrolysis cells

    KAUST Repository

    Rao, Hari Ananda

    2016-12-09

    Anode potential has been shown to be a critical factor in the rate of acetate removal in microbial electrolysis cells (MECs), but studies with fermentable substrates and set potentials are lacking. Here, we examined the impact of three different set anode potentials (SAPs; −0.25, 0, and 0.25 V vs. standard hydrogen electrode) on the electrochemical performance, electron flux to various sinks, and anodic microbial community structure in two-chambered MECs fed with propionate. Electrical current (49–71%) and CH4 (22.9–41%) were the largest electron sinks regardless of the potentials tested. Among the three SAPs tested, 0 V showed the highest electron flux to electrical current (71 ± 5%) and the lowest flux to CH4 (22.9 ± 1.2%). In contrast, the SAP of −0.25 V had the lowest electron flux to current (49 ± 6%) and the highest flux to CH4 (41.1 ± 2%). The most dominant genera detected on the anode of all three SAPs based on 16S rRNA gene sequencing were Geobacter, Smithella and Syntrophobacter, but their relative abundance varied among the tested SAPs. Microbial community analysis implies that complete degradation of propionate in all the tested SAPs was facilitated by syntrophic interactions between fermenters and Geobacter at the anode and ferementers and hydrogenotrophic methanogens in suspension.

  13. Comparative Metagenomic Analysis of Electrogenic Microbial Communities in Differentially Inoculated Swine Wastewater-Fed Microbial Fuel Cells

    Directory of Open Access Journals (Sweden)

    Irina V. Khilyas

    2017-01-01

    Full Text Available Bioelectrochemical systems such as microbial fuel cells (MFCs are promising new technologies for efficient removal of organic compounds from industrial wastewaters, including that generated from swine farming. We inoculated two pairs of laboratory-scale MFCs with sludge granules from a beer wastewater-treating anaerobic digester (IGBS or from sludge taken from the bottom of a tank receiving swine wastewater (SS. The SS-inoculated MFC outperformed the IGBS-inoculated MFC with regard to COD and VFA removal and electricity production. Using a metagenomic approach, we describe the microbial diversity of the MFC planktonic and anodic communities derived from the different inocula. Proteobacteria (mostly Deltaproteobacteria became the predominant phylum in both MFC anodic communities with amplification of the electrogenic genus Geobacter being the most pronounced. Eight dominant and three minor species of Geobacter were found in both MFC anodic communities. The anodic communities of the SS-inoculated MFCs had a higher proportion of Clostridium and Bacteroides relative to those of the IGBS-inoculated MFCs, which were enriched with Pelobacter. The archaeal populations of the SS- and IGBS-inoculated MFCs were dominated by Methanosarcina barkeri and Methanothermobacter thermautotrophicus, respectively. Our results show a long-term influence of inoculum type on the performance and microbial community composition of swine wastewater-treating MFCs.

  14. Strategies for Reducing the Start-up Operation of Microbial Electrochemical Treatments of Urban Wastewater

    Directory of Open Access Journals (Sweden)

    Zulema Borjas

    2015-12-01

    Full Text Available Microbial electrochemical technologies (METs constitute the core of a number of emerging technologies with a high potential for treating urban wastewater due to a fascinating reaction mechanism—the electron transfer between bacteria and electrodes to transform metabolism into electrical current. In the current work, we focus on the model electroactive microorganism Geobacter sulfurreducens to explore both the design of new start-up procedures and electrochemical operations. Our chemostat-grown plug and play cells, were able to reduce the start-up period by 20-fold while enhancing chemical oxygen demand (COD removal by more than 6-fold during this period. Moreover, a filter-press based bioreactor was successfully tested for both acetate-supplemented synthetic wastewater and real urban wastewater. This proof-of-concept pre-pilot treatment included a microbial electrolysis cell (MEC followed in time by a microbial fuel cell (MFC to finally generate electrical current of ca. 20 A·m−2 with a power of 10 W·m−2 while removing 42 g COD day−1·m−2. The effective removal of acetate suggests a potential use of this modular technology for treating acetogenic wastewater where Geobacter sulfurreducens outcompetes other organisms.

  15. Screen-Printed Electrodes: New Tools for Developing Microbial Electrochemistry at Microscale Level

    Directory of Open Access Journals (Sweden)

    Marta Estevez-Canales

    2015-11-01

    Full Text Available Microbial electrochemical technologies (METs have a number of potential technological applications. In this work, we report the use of screen-printed electrodes (SPEs as a tool to analyze the microbial electroactivity by using Geobacter sulfurreducens as a model microorganism. We took advantage of the small volume required for the assays (75 μL and the disposable nature of the manufactured strips to explore short-term responses of microbial extracellular electron transfer to conductive materials under different scenarios. The system proved to be robust for identifying the bioelectrochemical response, while avoiding complex electrochemical setups, not available in standard biotechnology laboratories. We successfully validated the system for characterizing the response of Geobacter sulfurreducens in different physiological states (exponential phase, stationary phase, and steady state under continuous culture conditions revealing different electron transfer responses. Moreover, a combination of SPE and G. sulfurreducens resulted to be a promising biosensor for quantifying the levels of acetate, as well as for performing studies in real wastewater. In addition, the potential of the technology for identifying electroactive consortia was tested, as an example, with a mixed population with nitrate-reducing capacity. We therefore present SPEs as a novel low-cost platform for assessing microbial electrochemical activity at the microscale level.

  16. Spatial distribution of bacterial communities on volumetric and planar anodes in single-chamber air-cathode microbial fuel cells

    KAUST Repository

    Vargas, Ignacio T.

    2013-05-29

    Pyrosequencing was used to characterize bacterial communities in air-cathode microbial fuel cells across a volumetric (graphite fiber brush) and a planar (carbon cloth) anode, where different physical and chemical gradients would be expected associated with the distance between anode location and the air cathode. As expected, the stable operational voltage and the coulombic efficiency (CE) were higher for the volumetric anode than the planar anode (0.57V and CE=22% vs. 0.51V and CE=12%). The genus Geobacter was the only known exoelectrogen among the observed dominant groups, comprising 57±4% of recovered sequences for the brush and 27±5% for the carbon-cloth anode. While the bacterial communities differed between the two anode materials, results showed that Geobacter spp. and other dominant bacterial groups were homogenously distributed across both planar and volumetric anodes. This lends support to previous community analysis interpretations based on a single biofilm sampling location in these systems. © 2013 Wiley Periodicals, Inc.

  17. Impact of acclimation methods on microbial communities and performance of anaerobic fluidized bed membrane bioreactors

    KAUST Repository

    Labarge, Nicole

    2016-10-17

    An anaerobic fluidized bed membrane bioreactor (AFMBR) is a new and effective method for energy-efficient treatment of low strength wastewater, but the factors that affect performance are not well known. Different inocula and acclimation methods of the granular activated carbon (GAC) used in the reactor were examined here to determine their impact on chemical oxygen demand (COD) removal and microbial community composition of domestic wastewater-fed AFMBRs. AFMBRs inoculated with anaerobic digester sludge (D) or domestic wastewater (W) and fed domestic wastewater, or inoculated with a microbiologically diverse anaerobic bog sediment and acclimated using methanol (M), all produced the same COD removal of 63 ± 12% using a diluted wastewater feed (100 ± 21 mg L−1 COD). However, an AFMBR with GAC inoculated with anaerobic digester sludge and acclimated using acetate (A) showed significantly increased wastewater COD removal to 84 ± 6%. In addition, feeding the AFMBR with the M-acclimated GAC with an acetate medium for one week subsequently increased COD removal to 70 ± 6%. Microbial communities enriched on the GAC included Geobacter, sulfur-reducing bacteria, Syntrophaceae, and Chlorobiaceae, with reactor A having the highest relative abundance of Geobacter. These results showed that acetate was the most useful substrate for acclimation of GAC communities, and GAC harbors unique communities relative to those in the AFMBR influent and recirculated solution.

  18. Metagenomic analyses reveal the involvement of syntrophic consortia in methanol/electricity conversion in microbial fuel cells.

    Directory of Open Access Journals (Sweden)

    Ayaka Yamamuro

    Full Text Available Methanol is widely used in industrial processes, and as such, is discharged in large quantities in wastewater. Microbial fuel cells (MFCs have the potential to recover electric energy from organic pollutants in wastewater; however, the use of MFCs to generate electricity from methanol has not been reported. In the present study, we developed single-chamber MFCs that generated electricity from methanol at the maximum power density of 220 mW m(-2 (based on the projected area of the anode. In order to reveal how microbes generate electricity from methanol, pyrosequencing of 16S rRNA-gene amplicons and Illumina shotgun sequencing of metagenome were conducted. The pyrosequencing detected in abundance Dysgonomonas, Sporomusa, and Desulfovibrio in the electrolyte and anode and cathode biofilms, while Geobacter was detected only in the anode biofilm. Based on known physiological properties of these bacteria, it is considered that Sporomusa converts methanol into acetate, which is then utilized by Geobacter to generate electricity. This speculation is supported by results of shotgun metagenomics of the anode-biofilm microbes, which reconstructed relevant catabolic pathways in these bacteria. These results suggest that methanol is anaerobically catabolized by syntrophic bacterial consortia with electrodes as electron acceptors.

  19. Metagenomic analyses reveal the involvement of syntrophic consortia in methanol/electricity conversion in microbial fuel cells.

    Science.gov (United States)

    Yamamuro, Ayaka; Kouzuma, Atsushi; Abe, Takashi; Watanabe, Kazuya

    2014-01-01

    Methanol is widely used in industrial processes, and as such, is discharged in large quantities in wastewater. Microbial fuel cells (MFCs) have the potential to recover electric energy from organic pollutants in wastewater; however, the use of MFCs to generate electricity from methanol has not been reported. In the present study, we developed single-chamber MFCs that generated electricity from methanol at the maximum power density of 220 mW m(-2) (based on the projected area of the anode). In order to reveal how microbes generate electricity from methanol, pyrosequencing of 16S rRNA-gene amplicons and Illumina shotgun sequencing of metagenome were conducted. The pyrosequencing detected in abundance Dysgonomonas, Sporomusa, and Desulfovibrio in the electrolyte and anode and cathode biofilms, while Geobacter was detected only in the anode biofilm. Based on known physiological properties of these bacteria, it is considered that Sporomusa converts methanol into acetate, which is then utilized by Geobacter to generate electricity. This speculation is supported by results of shotgun metagenomics of the anode-biofilm microbes, which reconstructed relevant catabolic pathways in these bacteria. These results suggest that methanol is anaerobically catabolized by syntrophic bacterial consortia with electrodes as electron acceptors.

  20. A Highly Efficient Mixed-culture Biofilm as Anodic Catalyst and Insights into Its Enhancement through Electrochemistry by Comparison with G. sulfurreducens

    International Nuclear Information System (INIS)

    Liu, Ying; Deng, Dandan; Lan, Xiaoji

    2015-01-01

    Highlights: • A mixed-culture biofilm with 68.6% higher current than Geobacter sulfurreducens was firstly reported, while G. sulfurreducens biofilm showed five-time higher apparent affinity than the mixed-culture. • The mixed-culture biofilm showed surface-controlled process, while diffusion-controlled process was obtained for G. sulfurreducens as at certain accelerating scan rates. • When the used medium was replaced with the fresh, decrease percentage of currents for both kinds of biofilms is similar (50%). • A suitable community will be an alternative for improving MFC performance. - Abstract: In this paper an efficient mixed-culture microbial biofilm with increased current density by 68.6% (1020.9 ± 47 μA cm −2 ) than that on typical culture of Geobacter sulfurreducens biofilm was firstly reported. The insights into the enhanced electricity-producing ability was investigated through evaluating the dependence of limiting current density on electroactive biomass coverage, replacing used growth medium, applying stirring and electron transfer kinetics. It was shown that the enhanced electricity generation ability of the mixed-culture biofilm is from population superiority of active molecules or electron shuttles from the biofilm. This work suggested that the optimized synergistic effect between interspecies in community could significantly improve electricity-producing performance than single strain. This study highlighted the potential synergistic role in special community on electricity generation capability

  1. Molecular Basis for Electron Flow Within Metal-and Electrode-Reducing Biofilms

    Energy Technology Data Exchange (ETDEWEB)

    Bond, Daniel R. [Univ. of Minnesota, Minneapolis, MN (United States)

    2016-11-01

    Electrochemical, spectral, genetic, and biochemical techniques were developed to reveal that a diverse suite of redox proteins and structural macromolecules outside the cell work together to move electrons long distances between Geobacter cells to metals and electrodes. In this project, we greatly expanded the known participants in the electron transfer pathway of Geobacter. For example, in addition to well-studied pili, polysaccharides contribute to anchoring, different cytochromes are required under different conditions, strategies change with redox potential, and the localization of these components can change depending on where cells are located in a biofilm. By inventing new electrodes compatible with real-time spectral measurements, we were able to visualize the redox status of biofilms in action, leading to a hypothesis that long-distance electron transfer is ultimately limiting in these systems and redox potentials change within biofilms. The goals of this project were met, as we were able to 1) identify new elements crucial to the expression, assembly and function of the extracellular electron transfer phenotype 2) expand spectral and electrochemical techniques to define the mechanism and route of electron transfer through the matrix, and 3) combine this knowledge to build the next generation of genetic tools for study of this complex process.

  2. The presence of hydrogenotrophic methanogens in the inoculum improves methane gas production in microbial electrolysis cells.

    KAUST Repository

    Siegert, Michael; Li, Xiu-Fen; Yates, Matthew D; Logan, Bruce E

    2014-01-01

    High current densities in microbial electrolysis cells (MECs) result from the predominance of various Geobacter species on the anode, but it is not known if archaeal communities similarly converge to one specific genus. MECs were examined here on the basis of maximum methane production and current density relative to the inoculum community structure. We used anaerobic digester (AD) sludge dominated by acetoclastic Methanosaeta, and an anaerobic bog sediment where hydrogenotrophic methanogens were detected. Inoculation using solids to medium ratio of 25% (w/v) resulted in the highest methane production rates (0.27 mL mL(-1) cm(-2), gas volume normalized by liquid volume and cathode projected area) and highest peak current densities (0.5 mA cm(-2)) for the bog sample. Methane production was independent of solid to medium ratio when AD sludge was used as the inoculum. 16S rRNA gene community analysis using pyrosequencing and quantitative PCR confirmed the convergence of Archaea to Methanobacterium and Methanobrevibacter, and of Bacteria to Geobacter, despite their absence in AD sludge. Combined with other studies, these findings suggest that Archaea of the hydrogenotrophic genera Methanobacterium and Methanobrevibacter are the most important microorganisms for methane production in MECs and that their presence in the inoculum improves the performance.

  3. Electrochemical evaluation of Ti/TiO{sub 2}-polyaniline anodes for microbial fuel cells using hypersaline microbial consortia for synthetic-wastewater treatment

    Energy Technology Data Exchange (ETDEWEB)

    Benetton, X.D.; Navarro-Avila, S.G. [Univ. Autonoma de Yucatan, Yucatan (Mexico). Biotecnologia y Bioingenieria; Carrera-Figueiras, C. [Univ. Autonoma de Yucatan, Yucatan (Mexico). Quimica Fundamental y Aplicada

    2010-07-01

    This paper described the development of a titanium (Ti/TiO{sub 2}) polyaniline composite electrode. The electrode was designed for use with a microbial fuel cell (MFC) that generated electricity through the microbial biodegradation of organic compounds. A modified NBAF medium was used with a 20 mM acetate as an electron donor and 53 mM fumarate as an electron acceptor for a period of 96 hours at 37 degrees C. Strains were cultured under strict anaerobic conditions. Two microbial cultures were used: (1) pure cultures of Geobacter sulfur-reducens; and (2) an uncharacterized stable microbial consortia isolated from hypersaline swamp sediments. The anodes were made with an emeraldine form of PANI deposited over Ti/TiO{sub 2} electrodes. Electrochemical impedance spectroscopy (EIS) monitoring was used to determine the open circuit potential of the MFC. Negative real impedances were obtained and reproduced in all systems studied with the Ti/TiO{sub 2}-PANI anodes. The highest power density was obtained using the Geobacter sulfur-reducens culture. Further research is needed to study the mechanisms that contribute to the occurrence of negative real impedances. 23 refs., 1 tab., 5 figs.

  4. Phase Preference by Active, Acetate-Utilizing Bacteria at the Rifle, CO Integrated Field Research Challenge Site

    Energy Technology Data Exchange (ETDEWEB)

    Kerkhof, L.; Williams, K.H.; Long, P.E.; McGuinness, L.

    2011-02-21

    Previous experiments at the Rifle, Colorado Integrated Field Research Challenge (IFRC) site demonstrated that field-scale addition of acetate to groundwater reduced the ambient soluble uranium concentration. In this report, sediment samples collected before and after acetate field addition were used to assess the active microbes via {sup 13}C acetate stable isotope probing on 3 phases [coarse sand, fines (8-approximately 150 {micro}m), groundwater (0.2-8 {micro}m)] over a 24-day time frame. TRFLP results generally indicated a stronger signal in {sup 13}C-DNA in the 'fines' fraction compared to the sand and groundwater. Before the field-scale acetate addition, a Geobacter-like group primarily synthesized {sup 13}C-DNA in the groundwater phase, an alpha Proteobacterium primarily grew on the fines/sands, and an Acinetobacter sp. and Decholoromonas-like OTU utilized much of the {sup 13}C acetate in both groundwater and particle-associated phases. At the termination of the field-scale acetate addition, the Geobacter-like species was active on the solid phases rather than the groundwater, while the other bacterial groups had very reduced newly synthesized DNA signal. These findings will help to delineate the acetate utilization patterns of bacteria in the field and can lead to improved methods for stimulating distinct microbial populations in situ.

  5. Using proteomic data to assess a genome-scale "in silico" model of metal reducing bacteria in the simulation of field-scale uranium bioremediation

    Science.gov (United States)

    Yabusaki, S.; Fang, Y.; Wilkins, M. J.; Long, P.; Rifle IFRC Science Team

    2011-12-01

    A series of field experiments in a shallow alluvial aquifer at a former uranium mill tailings site have demonstrated that indigenous bacteria can be stimulated with acetate to catalyze the conversion of hexavalent uranium in a groundwater plume to immobile solid-associated uranium in the +4 oxidation state. While this bioreduction of uranium has been shown to lower groundwater concentrations below actionable standards, a viable remediation methodology will need a mechanistic, predictive and quantitative understanding of the microbially-mediated reactions that catalyze the reduction of uranium in the context of site-specific processes, properties, and conditions. At the Rifle IFRC site, we are investigating the impacts on uranium behavior of pulsed acetate amendment, acetate-oxidizing iron and sulfate reducing bacteria, seasonal water table variation, spatially-variable physical (hydraulic conductivity, porosity) and geochemical (reactive surface area) material properties. The simulation of three-dimensional, variably saturated flow and biogeochemical reactive transport during a uranium bioremediation field experiment includes a genome-scale in silico model of Geobacter sp. to represent the Fe(III) terminal electron accepting process (TEAP). The Geobacter in silico model of cell-scale physiological metabolic pathways is comprised of hundreds of intra-cellular and environmental exchange reactions. One advantage of this approach is that the TEAP reaction stoichiometry and rate are now functions of the metabolic status of the microorganism. The linkage of in silico model reactions to specific Geobacter proteins has enabled the use of groundwater proteomic analyses to assess the accuracy of the model under evolving hydrologic and biogeochemical conditions. In this case, the largest predicted fluxes through in silico model reactions generally correspond to high abundances of proteins linked to those reactions (e.g. the condensation reaction catalyzed by the protein

  6. Polymorphism of proteins in selected slovak winter wheat genotypes using SDS-PAGE

    Directory of Open Access Journals (Sweden)

    Dana Miháliková

    2016-12-01

    Full Text Available Winter wheat is especially used for bread-making. The specific composition of the grain storage proteins and the representation of individual subunits determines the baking quality of wheat. The aim of this study was to analyze 15 slovak varieties of the winter wheat (Triticum aestivum L. based on protein polymorphism and to predict their technological quality. SDS-PAGE method by ISTA was used to separate glutenin protein subunits. Glutenins were separated into HMW-GS (15.13% and LMW-GS (65.89% on the basis of molecular weight in SDS-PAGE. At the locus Glu-A1 was found allele Null (53% of genotypes and allele 1 (47% of genotypes. The locus Glu-B1 was represented by the HMW-GS subunits 6+8 (33% of genotypes, 7+8 (27% of genotypes, 7+9 (40% of genotypes. At the locus Glu-D1 were detected two subunits, 2+12 (33% of genotypes and 5+10 (67% of genotypes which is correlated with good bread-making properties. The Glu – score was ranged from 4 (genotype Viglanka to 10 (genotypes Viola, Vladarka. According to the representation of individual glutenin subunits in samples, the dendrogram of genetic similarity was constructed. By the prediction of quality the results showed that the best technological quality was significant in the varieties Viola and Vladarka which are suitable for use in food processing.

  7. A low specific on-resistance SOI MOSFET with dual gates and a recessed drain

    International Nuclear Information System (INIS)

    Luo Xiao-Rong; Hu Gang-Yi; Zhang Zheng-Yuan; Luo Yin-Chun; Fan Ye; Wang Xiao-Wei; Fan Yuan-Hang; Cai Jin-Yong; Wang Pei; Zhou Kun

    2013-01-01

    A low specific on-resistance (R on,sp ) integrable silicon-on-insulator (SOI) metal-oxide semiconductor field-effect transistor (MOSFET) is proposed and investigated by simulation. The MOSFET features a recessed drain as well as dual gates, which consist of a planar gate and a trench gate extended to the buried oxide layer (BOX) (DGRD MOSFET). First, the dual gates form dual conduction channels, and the extended trench gate also acts as a field plate to improve the electric field distribution. Second, the combination of the trench gate and the recessed drain widens the vertical conduction area and shortens the current path. Third, the P-type top layer not only enhances the drift doping concentration but also modulates the surface electric field distributions. All of these sharply reduce R on,sp and maintain a high breakdown voltage (BV). The BV of 233 V and R on,sp of 4.151 mΩ·cm 2 (V GS = 15 V) are obtained for the DGRD MOSFET with 15-μm half-cell pitch. Compared with the trench gate SOI MOSFET and the conventional MOSFET, R on,sp of the DGRD MOSFET decreases by 36% and 33% with the same BV, respectively. The trench gate extended to the BOX synchronously acts as a dielectric isolation trench, simplifying the fabrication processes. (condensed matter: electronic structure, electrical, magnetic, and optical properties)

  8. Strategies for merging microbial fuel cell technologies in water desalination processes: Start-up protocol and desalination efficiency assessment

    Science.gov (United States)

    Borjas, Zulema; Esteve-Núñez, Abraham; Ortiz, Juan Manuel

    2017-07-01

    Microbial Desalination Cells constitute an innovative technology where microbial fuel cell and electrodialysis merge in the same device for obtaining fresh water from saline water with no energy-associated cost for the user. In this work, an anodic biofilm of the electroactive bacteria Geobacter sulfurreducens was able to efficiently convert the acetate present in synthetic waste water into electric current (j = 0.32 mA cm-2) able to desalinate water. .Moreover, we implemented an efficient start-up protocol where desalination up to 90% occurred in a desalination cycle (water production:0.308 L m-2 h-1, initial salinity: 9 mS cm-1, final salinity: osmosis (RO) or reverse electrodialysis.

  9. Microbial characterization and degradation of linear alkylbenzene sulfonate in an anaerobic reactor treating wastewater containing soap powder.

    Science.gov (United States)

    Carosia, Mariana Fronja; Okada, Dagoberto Yukio; Sakamoto, Isabel Kimiko; Silva, Edson Luiz; Varesche, Maria Bernadete Amâncio

    2014-09-01

    The aim of this study was to evaluate the removal of linear alkylbenzene sulfonate (LAS) in an anaerobic fluidized bed reactor (AFBR) treating wastewater containing soap powder as LAS source. At Stage I, the AFBR was fed with a synthetic substrate containing yeast extract and ethanol as carbon sources, and without LAS; at Stage II, soap powder was added to this synthetic substrate obtaining an LAS concentration of 14 ± 3 mg L(-1). The compounds of soap powder probably inhibited some groups of microorganisms, increasing the concentration of volatile fatty acids (VFA) from 91 to 143 mg HAc L(-1). Consequently, the LAS removal rate was 48 ± 10% after the 156 days of operation. By sequencing, 16S rRNA clones belonging to the phyla Proteobacteria and Synergistetes were identified in the samples taken at the end of the experiment, with a remarkable presence of Dechloromonas sp. and Geobacter sp. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. The performance of a thermophilic microbial fuel cell fed with synthesis gas.

    Science.gov (United States)

    Hussain, A; Mehta, P; Raghavan, V; Wang, H; Guiot, S R; Tartakovsky, B

    2012-08-10

    This study demonstrated electricity generation in a thermophilic microbial fuel cell (MFC) operated on synthesis gas (syngas) as the sole electron donor. At 50°C, a volumetric power output of 30-35 mWL(R)(-1) and a syngas conversion efficiency of 87-98% was achieved. The observed pathway of syngas conversion to electricity primarily consisted of a two-step process, where the carbon monoxide and hydrogen were first converted to acetate, which was then consumed by the anodophilic bacteria to produce electricity. A denaturing gradient gel electrophoresis (DGGE) analysis of the 16S rDNA revealed the presence of Geobacter species, Acetobacter, methanogens and several uncultured bacteria and archaea in the anodic chamber. Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.

  11. Metagenomic insight into methanogenic reactors promoting direct interspecies electron transfer via granular activated carbon.

    Science.gov (United States)

    Park, Jeong-Hoon; Park, Jong-Hun; Je Seong, Hoon; Sul, Woo Jun; Jin, Kang-Hyun; Park, Hee-Deung

    2018-07-01

    To provide insight into direct interspecies electron transfer via granular activated carbon (GAC), the effect of GAC supplementation on anaerobic digestion was evaluated. Compared to control samples, the GAC supplementation increased the total amount of methane production and its production rate by 31% and 72%, respectively. 16S rDNA sequencing analysis revealed a shift in the archaeal community composition; the Methanosarcina proportion decreased 17%, while the Methanosaeta proportion increased 5.6%. Metagenomic analyses based on shotgun sequencing demonstrated that the abundance of pilA and omcS genes belonging to Geobacter species decreased 69.4% and 29.4%, respectively. Furthermore, the analyses suggested a carbon dioxide reduction pathway rather than an acetate decarboxylation pathway for methane formation. Taken together, these results suggest that GAC improved methane production performance by shifting the microbial community and altering functional genes associated with direct interspecies electron transfer via conductive materials. Copyright © 2018 Elsevier Ltd. All rights reserved.

  12. Use of atomic force microscopy and transmission electron microscopy for correlative studies of bacterial capsules.

    Science.gov (United States)

    Stukalov, Oleg; Korenevsky, Anton; Beveridge, Terry J; Dutcher, John R

    2008-09-01

    Bacteria can possess an outermost assembly of polysaccharide molecules, a capsule, which is attached to their cell wall. We have used two complementary, high-resolution microscopy techniques, atomic force microscopy (AFM) and transmission electron microscopy (TEM), to study bacterial capsules of four different gram-negative bacterial strains: Escherichia coli K30, Pseudomonas aeruginosa FRD1, Shewanella oneidensis MR-4, and Geobacter sulfurreducens PCA. TEM analysis of bacterial cells using different preparative techniques (whole-cell mounts, conventional embeddings, and freeze-substitution) revealed capsules for some but not all of the strains. In contrast, the use of AFM allowed the unambiguous identification of the presence of capsules on all strains used in the present study, including those that were shown by TEM to be not encapsulated. In addition, the use of AFM phase imaging allowed the visualization of the bacterial cell within the capsule, with a depth sensitivity that decreased with increasing tapping frequency.

  13. A comparative evaluation of different types of microbial electrolysis desalination cells for malic acid production.

    Science.gov (United States)

    Liu, Guangli; Zhou, Ying; Luo, Haiping; Cheng, Xing; Zhang, Renduo; Teng, Wenkai

    2015-12-01

    The aim of this study was to investigate different microbial electrolysis desalination cells for malic acid production. The systems included microbial electrolysis desalination and chemical-production cell (MEDCC), microbial electrolysis desalination cell (MEDC) with bipolar membrane and anion exchange membrane (BP-A MEDC), MEDC with bipolar membrane and cation exchange membrane (BP-C MEDC), and modified microbial desalination cell (M-MDC). The microbial electrolysis desalination cells performed differently in terms of malic acid production and energy consumption. The MEDCC performed best with the highest malic acid production rate (18.4 ± 0.6 mmol/Lh) and the lowest energy consumption (0.35 ± 0.14 kWh/kg). The best performance of MEDCC was attributable to the neutral pH condition in the anode chamber, the lowest internal resistance, and the highest Geobacter percentage of the anode biofilm population among all the reactors. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Anolyte recycling enhanced bioelectricity generation of the buffer-free single-chamber air-cathode microbial fuel cell.

    Science.gov (United States)

    Ren, Yueping; Chen, Jinli; Shi, Yugang; Li, Xiufen; Yang, Na; Wang, Xinhua

    2017-11-01

    Anolyte acidification is an inevitable restriction for the bioelectricity generation of buffer-free microbial fuel cells (MFCs). In this work, acidification of the buffer-free KCl anolyte has been thoroughly eliminated through anolyte recycling. The accumulated HCO 3 - concentration in the recycled KCl anolyte was above 50mM, which played as natural buffer and elevated the anolyte pH to above 8. The maximum power density (P max ) increased from 322.9mWm -2 to 527.2mWm -2 , which is comparable with the phosphate buffered MFC. Besides Geobacter genus, the gradually increased anolyte pH and conductivity induced the growing of electrochemically active Geoalkalibacter genus, in the anode biofilm. Anolyte recycling is a feasible strategy to strengthen the self-buffering capacity of buffer-free MFCs, thoroughly eliminate the anolyte acidification and prominently enhance the electric power. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Insights from the Genomes of Microbes Thriving in Uranium-Enriched Sediments.

    Science.gov (United States)

    Sutcliffe, Brodie; Chariton, Anthony A; Harford, Andrew J; Hose, Grant C; Stephenson, Sarah; Greenfield, Paul; Midgley, David J; Paulsen, Ian T

    2018-05-01

    Elevated uranium dose (4 g kg -1 ) causes a shift in billabong sediment communities that result in the enrichment of five bacterial species. These taxa include Geobacter, Geothrix and Dyella species, as well as a novel-potentially predatory-Bacteroidetes species, and a new member of class Anaerolineae (Chloroflexi). Additionally, a population of methanogenic Methanocella species was also identified. Genomic reconstruction and metabolic examination of these taxa reveal a host of divergent life strategies and putative niche partitioning. Resistance-nodulation-division heavy metal efflux (RND-HME) transporters are implicated as potential uranium tolerance strategies among the bacterial taxa. Potential interactions, uranium tolerance and ecologically relevant catabolism are presented in a conceptual model of life in this environment.

  16. Perchlorate remediation using packed-bed bioreactors and electricity generation in microbial fuel cells (MFCs)

    Science.gov (United States)

    Min, Booki

    Two pilot-scale fixed bed bioreactors were operated in continuous mode in order to treat groundwater contaminated by perchlorate. The bioreactors were constructed and operated side-by-side at the Texas Street Well Facility in Redlands, California. Each reactor was packed with either sand or plastic media. A perchlorate-reducing bacterium, Dechlorosoma sp. KJ, was used to inoculate the bioreactors. Perchlorate was successfully removed down to a non-detectable level (microbial fuel cells (MFCs), which were run either in batch or continuous mode. In batch experiments, both a pure culture (Geobactor metallireducens) and a mixed culture (wastewater inoculum) were used as the biocatalyst, and acetate was added as substrate in the anode chamber of the MFC. Power output in a membrane MFC with either inoculum was essentially the same, with 40 +/- 1 mW/m2 for G. metallireducens and 38 +/- 1 mW/m2 for mixed culture. A different type of the MFC containing a salt bridge instead of a membrane system was examined to generate power using the same substrate and pure culture as used in the membrane MFC. Power output in the salt bridge MFC was 2.2 mW/m 2. It was found that the lower power output was directly attributed to the higher internal resistance of the salt bridge system (19920 +/- 50 O) in comparison with that of the membrane system (1286 +/- 1 O). Continuous electricity generation was examined in a flat plate microbial fuel cell (FPMFC) using domestic wastewater and specific organic substrates. The FPMFC, containing a combined electrode/proton exchange membrane (PEM), was initially acclimated for one month to domestic wastewater, and then was operated as a plug flow reactor system. Power density using domestic wastewater as a substrate was 72 +/- 1 mW/m2 at a liquid flow rate of 0.39 mL/min (1.1 hr hydraulic retention time, HRT), and COD removal was 42%. At a longer HRT of 4.0 hr, the COD removal increased to 79%, and power density was 43 mW/m2. Several organic compounds

  17. Cd Mobility in Anoxic Fe-Mineral-Rich Environments - Potential Use of Fe(III)-Reducing Bacteria in Soil Remediation

    Science.gov (United States)

    Muehe, E. M.; Adaktylou, I. J.; Obst, M.; Schröder, C.; Behrens, S.; Hitchcock, A. P.; Tylsizczak, T.; Michel, F. M.; Krämer, U.; Kappler, A.

    2014-12-01

    Agricultural soils are increasingly burdened with heavy metals such as Cd from industrial sources and impure fertilizers. Metal contaminants enter the food chain via plant uptake from soil and negatively affect human and environmental health. New remediation approaches are needed to lower soil metal contents. To apply these remediation techniques successfully, it is necessary to understand how soil microbes and minerals interact with toxic metals. Here we show that microbial Fe(III) reduction initially mobilizes Cd before its immobilization under anoxic conditions. To study how microbial Fe(III) reduction influences Cd mobility, we isolated a new Cd-tolerant, Fe(III)-reducing Geobacter sp. from a heavily Cd-contaminated soil. In lab experiments, this Geobacter strain first mobilized Cd from Cd-loaded Fe(III) hydroxides followed by precipitation of Cd-bearing mineral phases. Using Mössbauer spectroscopy and scanning electron microscopy, the original and newly formed Cd-containing Fe(II) and Fe(III) mineral phases, including Cd-Fe-carbonates, Fe-phosphates and Fe-(oxyhydr)oxides, were identified and characterized. Using energy-dispersive X-ray spectroscopy and synchrotron-based scanning transmission X-ray microscopy, Cd was mapped in the Fe(II) mineral aggregates formed during microbial Fe(III) reduction. Microbial Fe(III) reduction mobilizes Cd prior to its precipitation in Cd-bearing mineral phases. The mobilized Cd could be taken up by phytoremediating plants, resulting in a net removal of Cd from contaminated sites. Alternatively, Cd precipitation could reduce Cd bioavailability in the environment, causing less toxic effects to crops and soil microbiota. However, the stability and thus bioavailability of these newly formed Fe-Cd mineral phases needs to be assessed thoroughly. Whether phytoremediation or immobilization of Cd in a mineral with reduced Cd bioavailability are feasible mechanisms to reduce toxic effects of Cd in the environment remains to be

  18. Multiple redox states of multiheme cytochromes may enable bacterial response to changing redox environments

    Science.gov (United States)

    Arbour, T.; Wrighton, K. C.; Mullin, S. W.; Castelle, C.; Luef, B.; Gilbert, B.; Banfield, J. F.

    2013-12-01

    Multiheme c-type cytochromes (MHCs) are key components in electron-transport pathways that enable some microorganisms to transfer electron byproducts of metabolism to a variety of minerals. As a response to changes in mineral redox potential, microbial communities may shift their membership, or individual organisms may adjust protein expression. Alternatively, the ability to respond may be conferred by the innate characteristics of certain electron-transport-chain components. Here, we used potentiostat-controlled microbial fuel cells (MFCs) to measure the timescale of response to imposed changes in redox conditions, thus placing constraints on the importance of these different mechanisms. In the experiments, a solid electrode acts as an electron-accepting mineral whose redox potential can be precisely controlled. We inoculated duplicate MFCs with a sediment/groundwater mixture from an aquifer at Rifle, Colorado, supplied acetate as an electron donor, and obtained stable, mixed-species biofilms dominated by Geobacter and a novel Geobacter-related family. We poised the anode at potentials spanning the range of natural Fe(III)-reduction, then performed cyclic voltammetry (CV) to characterize the overall biofilm redox signature. The apparent biofilm midpoint potential shifted directly with anode set potential when the latter was changed within the range from about -250 to -50 mV vs. SHE. Following a jump in set potential by 200 mV, the CV-midpoint shift by ~100 mV over a timescale of ~30 minutes to a few hours, depending on the direction of the potential change. The extracellular electron transfer molecules, whose overall CV signature is very similar to those of purified MHCs, appear to span a broad redox range (~200 mV), supporting the hypothesis that MHCs confer substantial redox flexibility. This flexibility may be a principle reason for the abundance of MHCs expressed by microorganisms capable of extracellular electron transfer to minerals.

  19. Exoelectrogenic biofilm as a template for sustainable formation of a catalytic mesoporous structure

    KAUST Repository

    Yates, Matthew D.

    2014-06-04

    © 2014 Wiley Periodicals, Inc. Actively respiring biofilms of Geobacter sulfurreducens were used as a biotemplate to form a palladium mesoporous layer directly on an electrode surface. Cells and proteins within the biofilm acted as the reductant and stabilizer to facilitate the reduction, dispersion, and attachment of palladium nanoparticles to the electrode surface without using synthetic chemicals. © 2014 Wiley Periodicals, Inc. Mesoporous structures can increase catalytic activity by maximizing the ratio of surface area to volume, but current synthesis techniques utilize expensive polymers and toxic chemicals. A Geobacter sulfurreducens biofilm was used as a sustainable template to form mesoporous Pd structures while eliminating the need for synthetic chemicals. The bulk of the biofilm material was removed by thermal treatments after nanoparticle formation, producing a catalytic Pd mesoporous (pore size 9.7±0.1nm) structure attached to the graphite electrode with a 1.5-2μm thick backbone composed of nanoparticles (~200nm). A control electrode electrochemically plated with Pd in the absence of a biofilm exhibited a variable planar Pd base (~0.5-3μm thick) with sporadic Pd extrusions (~2μm across, 1-5μm tall) from the surface. The biotemplated mesoporous structure produced 15-20% higher stable current densities during H2 oxidation tests than the electrochemically plated control electrode, even though 30% less Pd was present in the biotemplated catalyst. These results indicate that electroactive biofilms can be used as a sustainable base material to produce nanoporous structures without the need for synthetic polymers. Biotechnol. Bioeng. 2014;111: 2349-2354.

  20. Species and Scale Dependence of Bacterial Motion Dynamics

    Science.gov (United States)

    Sund, N. L.; Yang, X.; Parashar, R.; Plymale, A.; Hu, D.; Kelly, R.; Scheibe, T. D.

    2017-12-01

    Many metal reducing bacteria are motile with their motion characteristics described by run-and-tumble behavior exhibiting series of flights (jumps) and waiting (residence) time spanning a wide range of values. Accurate models of motility allow for improved design and evaluation of in-situ bioremediation in the subsurface. While many bioremediation models neglect the motion of the bacteria, others treat motility using an advection dispersion equation, which assumes that the motion of the bacteria is Brownian.The assumption of Brownian motion to describe motility has enormous implications on predictive capabilities of bioremediation models, yet experimental evidence of this assumption is mixed [1][2][3]. We hypothesize that this is due to the species and scale dependence of the motion dynamics. We test our hypothesis by analyzing videos of motile bacteria of five different species in open domains. Trajectories of individual cells ranging from several seconds to few minutes in duration are extracted in neutral conditions (in the absence of any chemical gradient). The density of the bacteria is kept low so that the interaction between the bacteria is minimal. Preliminary results show a transition from Fickian (Brownian) to non-Fickian behavior for one species of bacteria (Pelosinus) and persistent Fickian behavior of another species (Geobacter).Figure: Video frames of motile bacteria with the last 10 seconds of their trajectories drawn in red. (left) Pelosinus and (right) Geobacter.[1] Ariel, Gil, et al. "Swarming bacteria migrate by Lévy Walk." Nature Communications 6 (2015).[2] Saragosti, Jonathan, Pascal Silberzan, and Axel Buguin. "Modeling E. coli tumbles by rotational diffusion. Implications for chemotaxis." PloS one 7.4 (2012): e35412.[3] Wu, Mingming, et al. "Collective bacterial dynamics revealed using a three-dimensional population-scale defocused particle tracking technique." Applied and Environmental Microbiology 72.7 (2006): 4987-4994.

  1. Reactor performance in terms of COD and nitrogen removal and bacterial community structure of a three-stage rotating bioelectrochemical contactor

    KAUST Repository

    Sayess, Rassil R.

    2013-02-01

    Integrating microbial fuel cell (MFC) into rotating biological contactor (RBC) creates an opportunity for enhanced removal of COD and nitrogen coupled with energy generation from wastewater. In this study, a three-stage rotating bioelectrochemical contactor (referred to as RBC-MFC unit) integrating MFC with RBC technology was constructed for simultaneous removal of carbonaceous and nitrogenous compounds and electricity generation from a synthetic medium containing acetate and ammonium. The performance of the RBC-MFC unit was compared to a control reactor (referred to as RBC unit) that was operated under the same conditions but without current generation (i.e. open-circuit mode). The effect of hydraulic loading rate (HLR) and COD/N ratio on the performance of the two units was investigated. At low (3.05 gCOD g-1N) and high COD/N ratio (6.64 gCOD g-1N), both units achieved almost similar COD and ammonia-nitrogen removal. However, the RBC-MFC unit achieved significantly higher denitrification and nitrogen removal compared to the RBC unit indicating improved denitrification at the cathode due to current flow. The average voltage under 1000 Ω external resistance ranged between 0.03 and 0.30 V and between 0.02 and 0.21 V for stages 1 and 2 of the RBC-MFC unit. Pyrosequencing analysis of bacterial 16S rRNA gene revealed high bacterial diversity at the anode and cathode of both units. Genera that play a role in nitrification (Nitrospira; Nitrosomonas), denitrification (Comamonas; Thauera) and electricity generation (Geobacter) were identified at the electrodes. Geobacter was only detected on the anode of the RBC-MFC unit. Nitrifiers and denitrifiers were more abundant in the RBC-MFC unit compared to the RBC unit and were largely present on the cathode of both units suggesting that most of the nitrogen removal occurred at the cathode. © 2012 Elsevier Ltd.

  2. Exoelectrogenic biofilm as a template for sustainable formation of a catalytic mesoporous structure

    KAUST Repository

    Yates, Matthew D.; Cusick, Roland D.; Ivanov, Ivan; Logan, Bruce E.

    2014-01-01

    © 2014 Wiley Periodicals, Inc. Actively respiring biofilms of Geobacter sulfurreducens were used as a biotemplate to form a palladium mesoporous layer directly on an electrode surface. Cells and proteins within the biofilm acted as the reductant and stabilizer to facilitate the reduction, dispersion, and attachment of palladium nanoparticles to the electrode surface without using synthetic chemicals. © 2014 Wiley Periodicals, Inc. Mesoporous structures can increase catalytic activity by maximizing the ratio of surface area to volume, but current synthesis techniques utilize expensive polymers and toxic chemicals. A Geobacter sulfurreducens biofilm was used as a sustainable template to form mesoporous Pd structures while eliminating the need for synthetic chemicals. The bulk of the biofilm material was removed by thermal treatments after nanoparticle formation, producing a catalytic Pd mesoporous (pore size 9.7±0.1nm) structure attached to the graphite electrode with a 1.5-2μm thick backbone composed of nanoparticles (~200nm). A control electrode electrochemically plated with Pd in the absence of a biofilm exhibited a variable planar Pd base (~0.5-3μm thick) with sporadic Pd extrusions (~2μm across, 1-5μm tall) from the surface. The biotemplated mesoporous structure produced 15-20% higher stable current densities during H2 oxidation tests than the electrochemically plated control electrode, even though 30% less Pd was present in the biotemplated catalyst. These results indicate that electroactive biofilms can be used as a sustainable base material to produce nanoporous structures without the need for synthetic polymers. Biotechnol. Bioeng. 2014;111: 2349-2354.

  3. Influence of Oxygen and Nitrate on Fe (Hydr)oxide Mineral Transformation and Soil Microbial Communities during Redox Cycling.

    Science.gov (United States)

    Mejia, Jacqueline; Roden, Eric E; Ginder-Vogel, Matthew

    2016-04-05

    Oscillations between reducing and oxidizing conditions are observed at the interface of anaerobic/oxic and anaerobic/anoxic environments, and are often stimulated by an alternating flux of electron donors (e.g., organic carbon) and electron acceptors (e.g., O2 and NO3(-)). In iron (Fe) rich soils and sediments, these oscillations may stimulate the growth of both Fe-reducing bacteria (FeRB) and Fe-oxidizing bacteria (FeOB), and their metabolism may induce cycling between Fe(II) and Fe(III), promoting the transformation of Fe (hydr)oxide minerals. Here, we examine the mineralogical evolution of lepidocrocite and ferrihydrite, and the adaptation of a natural microbial community to alternating Fe-reducing (anaerobic with addition of glucose) and Fe-oxidizing (with addition of nitrate or air) conditions. The growth of FeRB (e.g., Geobacter) is stimulated under anaerobic conditions in the presence of glucose. However, the abundance of these organisms depends on the availability of Fe(III) (hydr)oxides. Redox cycling with nitrate results in decreased Fe(II) oxidation thereby decreasing the availability of Fe(III) for FeRB. Additionally, magnetite is detected as the main product of both lepidocrocite and ferrihydrite reduction. In contrast, introduction of air results in increased Fe(II) oxidation, increasing the availability of Fe(III) and the abundance of Geobacter. In the lepidocrocite reactors, Fe(II) oxidation by dissolved O2 promotes the formation of ferrihydrite and lepidocrocite, whereas in the ferrihydrite reactors we observe a decrease in magnetite stoichiometry (e.g., oxidation). Understanding Fe (hydr)oxide transformation under environmentally relevant redox cycling conditions provides insight into nutrient availability and transport, contaminant mobility, and microbial metabolism in soils and sediments.

  4. Phylogenetic diversity of dissimilatory ferric iron reducers in paddy soil of Hunan, South China

    Energy Technology Data Exchange (ETDEWEB)

    Wang Xin-Jun [State Key Lab. of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, BJ (China); Graduate Univ., Chinese Academy of Sciences, BJ (China); Yang Jing; Chen Xue-Ping; Sun Guo-Xin [State Key Lab. of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, BJ (China); Zhu Yong-Guan [State Key Lab. of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, BJ (China); Key Lab. of Urban Environment and Health, Inst. of Urban Environment, Chinese Academy of Sciences, Xiamen (China)

    2009-12-15

    Purpose: Dissimilatory iron-reducing bacteria have been described by both culture-dependent and -independent methods in various environments, including freshwater, marine sediments, natural wetlands, and contaminated aquifers. However, little is known about iron-reducing microbial communities in paddy soils. The goal of this study was to characterize iron-reducing microbial communities in paddy soil. Moreover, the effect of dissolved and solid-phase iron (III) species on the iron-reducing microbial communities was also investigated by enrichment cultures. Methods: Ferric citrate and ferrihydrite were used respectively to set up enrichment cultures of dissimilatory ironreducing microorganisms using 1% inoculum of soil samples, and the iron reduction was measured. Moreover, bacterial DNA was extracted and 16S rRNA genes were PCR-amplified, and subsequently analyzed by the clone library and terminal restriction fragment length polymorphism (T-RFLP). Results: Phylogenetic analysis of 16S rRNA gene sequences extracted from the enrichment cultures revealed that Bradyrhizobium, Bacteroides, Clostridium and Ralstonia species were the dominant bacteria in the ferric citrate enrichment. However, members of the genera Clostridium, Bacteroides, and Geobacter were the dominant micro-organisms in the ferrihydrite enrichment. Analysis of enrichment cultures by T-RFLP strongly supported the cloning and sequencing results. Conclusions: The present study demonstrated that dissimilatory iron-reducing consortia in As-contaminated paddy soil are phylogenetically diverse. Moreover, iron (III) sources as a key factor have a strong effect on the iron (III)-reducing microbial community structure and relative abundance in the enrichments. In addition, Geobacter species are selectively enriched by ferrihydrite enrichment cultures. (orig.)

  5. Iron-reducing bacteria accumulate ferric oxyhydroxide nanoparticle aggregates that may support planktonic growth.

    Science.gov (United States)

    Luef, Birgit; Fakra, Sirine C; Csencsits, Roseann; Wrighton, Kelly C; Williams, Kenneth H; Wilkins, Michael J; Downing, Kenneth H; Long, Philip E; Comolli, Luis R; Banfield, Jillian F

    2013-02-01

    Iron-reducing bacteria (FeRB) play key roles in anaerobic metal and carbon cycling and carry out biogeochemical transformations that can be harnessed for environmental bioremediation. A subset of FeRB require direct contact with Fe(III)-bearing minerals for dissimilatory growth, yet these bacteria must move between mineral particles. Furthermore, they proliferate in planktonic consortia during biostimulation experiments. Thus, a key question is how such organisms can sustain growth under these conditions. Here we characterized planktonic microbial communities sampled from an aquifer in Rifle, Colorado, USA, close to the peak of iron reduction following in situ acetate amendment. Samples were cryo-plunged on site and subsequently examined using correlated two- and three-dimensional cryogenic transmission electron microscopy (cryo-TEM) and scanning transmission X-ray microscopy (STXM). The outer membranes of most cells were decorated with aggregates up to 150 nm in diameter composed of ∼3 nm wide amorphous, Fe-rich nanoparticles. Fluorescent in situ hybridization of lineage-specific probes applied to rRNA of cells subsequently imaged via cryo-TEM identified Geobacter spp., a well-studied group of FeRB. STXM results at the Fe L(2,3) absorption edges indicate that nanoparticle aggregates contain a variable mixture of Fe(II)-Fe(III), and are generally enriched in Fe(III). Geobacter bemidjiensis cultivated anaerobically in the laboratory on acetate and hydrous ferric oxyhydroxides also accumulated mixed-valence nanoparticle aggregates. In field-collected samples, FeRB with a wide variety of morphologies were associated with nano-aggregates, indicating that cell surface Fe(III) accumulation may be a general mechanism by which FeRB can grow while in planktonic suspension.

  6. Influence of dissimilatory metal reduction on fate of organic and metal contaminants in the subsurface

    Science.gov (United States)

    Lovley, Derek R.; Anderson, Robert T.

    Dissimilatory Fe(III)-reducing microorganisms have the ability to destroy organic contaminants under anaerobic conditions by oxidizing them to carbon dioxide. Some Fe(III)-reducing microorganisms can also reductively dechlorinate chlorinated contaminants. Fe(III)-reducing microorganisms can reduce a variety of contaminant metals and convert them from soluble forms to forms that are likely to be immobilized in the subsurface. Studies in petroleum-contaminated aquifers have demonstrated that Fe(III)-reducing microorganisms can be effective agents in removing aromatic hydrocarbons from groundwater under anaerobic conditions. Laboratory studies have demonstrated the potential for Fe(III)-reducing microorganisms to remove uranium from contaminated groundwaters. The activity of Fe(III)-reducing microorganisms can be stimulated in several ways to enhance organic contaminant oxidation and metal reduction. Molecular analyses in both field and laboratory studies have demonstrated that microorganisms of the genus Geobacter become dominant members of the microbial community when Fe(III)-reducing conditions develop as the result of organic contamination, or when Fe(III) reduction is artificially stimulated. These results suggest that further understanding of the ecophysiology of Geobacter species would aid in better prediction of the natural attenuation of organic contaminants under anaerobic conditions and in the design of strategies for the bioremediation of subsurface metal contamination. Des micro-organismes simulant la réduction du fer ont la capacité de détruire des polluants organiques dans des conditions anérobies en les oxydant en dioxyde de carbone. Certains micro-organismes réducteurs de fer peuvent aussi dé-chlorer par réduction des polluants chlorés. Des micro-organismes réducteurs de fer peuvent réduire tout un ensemble de métaux polluants et les faire passer de formes solubles à des formes qui sont susceptibles d'être immobilisées dans le milieu

  7. Shipborne LiDAR system for coastal change monitoring

    Science.gov (United States)

    Kim, chang hwan; Park, chang hong; Kim, hyun wook; hyuck Kim, won; Lee, myoung hoon; Park, hyeon yeong

    2016-04-01

    Coastal areas, used as human utilization areas like leisure space, medical care, ports and power plants, etc., are regions that are continuously changing and interconnected with oceans and land and the sea level has risen by about 8cm (1.9mm / yr) due to global warming from 1964 year to 2006 year in Korea. Coastal erosion due to sea-level rise has caused the problem of marine ecosystems and loss of tourism resources, etc. Regular monitoring of coastal erosion is essential at key locations with such volatility. But the survey method of land mobile LiDAR (light detection and ranging) system has much time consuming and many restrictions. For effective monitoring beach erosion, KIOST (Korea Institute of Ocean Science & Technology) has constructed a shipborne mobile LiDAR system. The shipborne mobile LiDAR system comprised a land mobile LiDAR (RIEGL LMS-420i), an INS (inertial navigation system, MAGUS Inertial+), a RTKGPS (LEICA GS15 GS25), and a fixed platform. The shipborne mobile LiDAR system is much more effective than a land mobile LiDAR system in the measuring of fore shore areas without shadow zone. Because the vessel with the shipborne mobile LiDAR system is continuously moved along the shoreline, it is possible to efficiently survey a large area in a relatively short time. Effective monitoring of the changes using the constructed shipborne mobile LiDAR system for seriously eroded coastal areas will be able to contribute to coastal erosion management and response.

  8. Biostimulation of Iron Reduction and Uranium Immobilization: Microbial and Mineralogical Controls

    International Nuclear Information System (INIS)

    Joel E. Kostka; Lainie Petrie; Nadia North; David L. Balkwill; Joseph W. Stucki; Lee Kerkhof

    2004-01-01

    change in microbial community composition of FRC sediments during in situ biostimulation in single well push-pull tests. Microbial communities were stimulated in the acidic subsurface via pH neutralization and addition of electron donor to wells. Examination of sediment chemistry in cores sampled immediately adjacent to treated wells revealed that sediment pH increased substantially (by 1-2 pH units), while nitrate was largely depleted. Following the in situ biostimulation, previously cultured metal-reducing (delta)-Proteobacteria 16S rRNA gene sequences substantially increased from 5% to nearly 40% of clone libraries. Quantitative PCR revealed that Geobacter-type 16S rRNA gene sequences increased in biostimulated sediments by one to two orders of magnitude at two of the four sites tested, thereby corroborating information obtained from clone libraries, and indicating that members of the (delta)-Proteobacteria (including Anaeromyxobacter dehalogenans-related and Geobacter-related organisms) are important metal-reducing bacteria in FRC.

  9. Effect of Start-Up Strategies and Electrode Materials on Carbon Dioxide Reduction on Biocathodes.

    Science.gov (United States)

    Saheb-Alam, Soroush; Singh, Abhijeet; Hermansson, Malte; Persson, Frank; Schnürer, Anna; Wilén, Britt-Marie; Modin, Oskar

    2018-02-15

    The enrichment of CO 2 -reducing microbial biocathodes is challenging. Previous research has shown that a promising approach could be to first enrich bioanodes and then lower the potential so the electrodes are converted into biocathodes. However, the effect of such a transition on the microbial community on the electrode has not been studied. The goal of this study was thus to compare the start-up of biocathodes from preenriched anodes with direct start-up from bare electrodes and to investigate changes in microbial community composition. The effect of three electrode materials on the long-term performance of the biocathodes was also investigated. In this study, preenrichment of acetate-oxidizing bioanodes did not facilitate the start-up of biocathodes. It took about 170 days for the preenriched electrodes to generate substantial cathodic current, compared to 83 days for the bare electrodes. Graphite foil and carbon felt cathodes produced higher current at the beginning of the experiment than did graphite rods. However, all electrodes produced similar current densities at the end of the over 1-year-long study (2.5 A/m 2 ). Methane was the only product detected during operation of the biocathodes. Acetate was the only product detected after inhibition of the methanogens. Microbial community analysis showed that Geobacter sp. dominated the bioanodes. On the biocathodes, the Geobacter sp. was succeeded by Methanobacterium spp., which made up more than 80% of the population. After inhibition of the methanogens, Acetobacterium sp. became dominant on the electrodes (40% relative abundance). The results suggested that bioelectrochemically generated H 2 acted as an electron donor for CO 2 reduction. IMPORTANCE In microbial electrochemical systems, living microorganisms function as catalysts for reactions on the anode and/or the cathode. There is a variety of potential applications, ranging from wastewater treatment and biogas generation to production of chemicals. Systems

  10. Improved Understanding of Microbial Iron and Sulfate Reduction Through a Combination of Bottom-up and Top-down Functional Proteomics Assays

    Energy Technology Data Exchange (ETDEWEB)

    Richardson, Ruth [Cornell Univ., Ithaca, NY (United States)

    2016-02-28

    Our overall goal was to improve the understanding of microbial iron and sulfate reduction by evaluating a diverse iron and sulfate reducing organisms utilizing a multi-omics approach combining “top-down” and “bottom-up” omics methodologies. We initiated one of the first combined comparative genomics, shotgun proteomics, RTqPCR, and heterologous expression studies in pursuit of our project objectives. Within the first year of this project, we created a new bioinformatics tool for ortholog identification (“SPOCS”). SPOCS is described in our publication, Curtis et al., 2013. Using this tool we were able to identify conserved orthologous groups across diverse iron and sulfate reducing microorganisms from Firmicutes, gamma-proteobacteria and delta-proteobacteria. For six iron and sulfate reducers we also performed shotgun proteomics (“bottom-up” proteomics including accurate mass and time (AMT) tag and iTRAQ approaches). Cultures include Gram (-) and Gram (+) microbes. Gram (-) were: Geobacter sulfureducens (grown on iron citrate and fumarate), Geobacter bemidjiensis (grown on iron citrate and fumarate), Shewanella oneidiensis (grown on iron citrate and fumarate) and Anaeromyxobacter dehalogenans (grown on iron citrate and fumarate). Although all cultures grew on insoluble iron, the iron precipitates interfered with protein extraction and analysis; which remains a major challenge for researchers in disparate study systems. Among the Gram (-) organisms studied, Anaeromyxobacter dehalogenans remains the most poorly characterized. Yet, it is arguably the most versatile organisms we studied. In this work we have used comparative proteomics to hypothesize which two of the dozens of predicted c-type cytochromes within Anaeromyxobacter dehalogenans may be directly involved in soluble iron reduction. Unfortunately, heterologous expression of these Anaeromyxobacter dehalogenans ctype cytochromes led to poor protein production and/or formation of inclusion bodies

  11. Ferrihydrite-associated organic matter (OM stimulates reduction by Shewanella oneidensis MR-1 and a complex microbial consortia

    Directory of Open Access Journals (Sweden)

    R. E. Cooper

    2017-11-01

    Full Text Available The formation of Fe(III oxides in natural environments occurs in the presence of natural organic matter (OM, resulting in the formation of OM–mineral complexes that form through adsorption or coprecipitation processes. Thus, microbial Fe(III reduction in natural environments most often occurs in the presence of OM–mineral complexes rather than pure Fe(III minerals. This study investigated to what extent does the content of adsorbed or coprecipitated OM on ferrihydrite influence the rate of Fe(III reduction by Shewanella oneidensis MR-1, a model Fe(III-reducing microorganism, in comparison to a microbial consortium extracted from the acidic, Fe-rich Schlöppnerbrunnen fen. We found that increased OM content led to increased rates of microbial Fe(III reduction by S. oneidensis MR-1 in contrast to earlier findings with the model organism Geobacter bremensis. Ferrihydrite–OM coprecipitates were reduced slightly faster than ferrihydrites with adsorbed OM. Surprisingly, the complex microbial consortia stimulated by a mixture of electrons donors (lactate, acetate, and glucose mimics S. oneidensis under the same experimental Fe(III-reducing conditions suggesting similar mechanisms of electron transfer whether or not the OM is adsorbed or coprecipitated to the mineral surfaces. We also followed potential shifts of the microbial community during the incubation via 16S rRNA gene sequence analyses to determine variations due to the presence of adsorbed or coprecipitated OM–ferrihydrite complexes in contrast to pure ferrihydrite. Community profile analyses showed no enrichment of typical model Fe(III-reducing bacteria, such as Shewanella or Geobacter sp., but an enrichment of fermenters (e.g., Enterobacteria during pure ferrihydrite incubations which are known to use Fe(III as an electron sink. Instead, OM–mineral complexes favored the enrichment of microbes including Desulfobacteria and Pelosinus sp., both of which can utilize lactate and

  12. Ferrihydrite-associated organic matter (OM) stimulates reduction by Shewanella oneidensis MR-1 and a complex microbial consortia

    Science.gov (United States)

    Cooper, Rebecca Elizabeth; Eusterhues, Karin; Wegner, Carl-Eric; Totsche, Kai Uwe; Küsel, Kirsten

    2017-11-01

    The formation of Fe(III) oxides in natural environments occurs in the presence of natural organic matter (OM), resulting in the formation of OM-mineral complexes that form through adsorption or coprecipitation processes. Thus, microbial Fe(III) reduction in natural environments most often occurs in the presence of OM-mineral complexes rather than pure Fe(III) minerals. This study investigated to what extent does the content of adsorbed or coprecipitated OM on ferrihydrite influence the rate of Fe(III) reduction by Shewanella oneidensis MR-1, a model Fe(III)-reducing microorganism, in comparison to a microbial consortium extracted from the acidic, Fe-rich Schlöppnerbrunnen fen. We found that increased OM content led to increased rates of microbial Fe(III) reduction by S. oneidensis MR-1 in contrast to earlier findings with the model organism Geobacter bremensis. Ferrihydrite-OM coprecipitates were reduced slightly faster than ferrihydrites with adsorbed OM. Surprisingly, the complex microbial consortia stimulated by a mixture of electrons donors (lactate, acetate, and glucose) mimics S. oneidensis under the same experimental Fe(III)-reducing conditions suggesting similar mechanisms of electron transfer whether or not the OM is adsorbed or coprecipitated to the mineral surfaces. We also followed potential shifts of the microbial community during the incubation via 16S rRNA gene sequence analyses to determine variations due to the presence of adsorbed or coprecipitated OM-ferrihydrite complexes in contrast to pure ferrihydrite. Community profile analyses showed no enrichment of typical model Fe(III)-reducing bacteria, such as Shewanella or Geobacter sp., but an enrichment of fermenters (e.g., Enterobacteria) during pure ferrihydrite incubations which are known to use Fe(III) as an electron sink. Instead, OM-mineral complexes favored the enrichment of microbes including Desulfobacteria and Pelosinus sp., both of which can utilize lactate and acetate as an electron

  13. Improved performance of the microbial electrolysis desalination and chemical-production cell with enlarged anode and high applied voltages.

    Science.gov (United States)

    Ye, Bo; Luo, Haiping; Lu, Yaobin; Liu, Guangli; Zhang, Renduo; Li, Xiao

    2017-11-01

    The aim of this study was to improve performance of the microbial electrolysis desalination and chemical-production cell (MEDCC) using enlarged anode and high applied voltages. MEDCCs with anode lengths of 9 and 48cm (i.e., the 9cm-anode MEDCC and 48cm-anode MEDCC, respectively) were tested under different voltages (1.2-3.0V). Our results demonstrated for the first time that the MEDCC could maintain high performance even under the applied voltage higher than that for water dissociation (i.e., 1.8V). Under the applied voltage of 2.5V, the maximum current density in the 48cm-anode MEDCC reached 32.8±2.6A/m 2 , which is one of the highest current densities reported so far in the bioelectrochemical system (BES). The relative abundance of Geobacter was changed along the anode length. Our results show the great potential of the BES with enlarged anode and high applied voltages. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. The biogeochemical fate of nickel during microbial ISA degradation; implications for nuclear waste disposal.

    Science.gov (United States)

    Kuippers, Gina; Boothman, Christopher; Bagshaw, Heath; Ward, Michael; Beard, Rebecca; Bryan, Nicholas; Lloyd, Jonathan R

    2018-06-08

    Intermediate level radioactive waste (ILW) generally contains a heterogeneous range of organic and inorganic materials, of which some are encapsulated in cement. Of particular concern are cellulosic waste items, which will chemically degrade under the conditions predicted during waste disposal, forming significant quantities of isosaccharinic acid (ISA), a strongly chelating ligand. ISA therefore has the potential to increase the mobility of a wide range of radionuclides via complex formation, including Ni-63 and Ni-59. Although ISA is known to be metabolized by anaerobic microorganisms, the biodegradation of metal-ISA complexes remains unexplored. This study investigates the fate of a Ni-ISA complex in Fe(III)-reducing enrichment cultures at neutral pH, representative of a microbial community in the subsurface. After initial sorption of Ni onto Fe(III)oxyhydroxides, microbial ISA biodegradation resulted in >90% removal of the remaining Ni from solution when present at 0.1 mM, whereas higher concentrations of Ni proved toxic. The microbial consortium associated with ISA degradation was dominated by close relatives to Clostridia and Geobacter species. Nickel was preferentially immobilized with trace amounts of biogenic amorphous iron sulfides. This study highlights the potential for microbial activity to help remove chelating agents and radionuclides from the groundwater in the subsurface geosphere surrounding a geodisposal facility.

  15. Evolution from a respiratory ancestor to fill syntrophic and fermentative niches: comparative fenomics of six Geobacteraceae species

    Directory of Open Access Journals (Sweden)

    Lovley Derek R

    2009-03-01

    Full Text Available Abstract Background The anaerobic degradation of organic matter in natural environments, and the biotechnical use of anaerobes in energy production and remediation of subsurface environments, both require the cooperative activity of a diversity of microorganisms in different metabolic niches. The Geobacteraceae family contains members with three important anaerobic metabolisms: fermentation, syntrophic degradation of fermentation intermediates, and anaerobic respiration. Results In order to learn more about the evolution of anaerobic microbial communities, the genome sequences of six Geobacteraceae species were analyzed. The results indicate that the last common Geobacteraceae ancestor contained sufficient genes for anaerobic respiration, completely oxidizing organic compounds with the reduction of external electron acceptors, features that are still retained in modern Geobacter and Desulfuromonas species. Evolution of specialization for fermentative growth arose twice, via distinct lateral gene transfer events, in Pelobacter carbinolicus and Pelobacter propionicus. Furthermore, P. carbinolicus gained hydrogenase genes and genes for ferredoxin reduction that appear to permit syntrophic growth via hydrogen production. The gain of new physiological capabilities in the Pelobacter species were accompanied by the loss of several key genes necessary for the complete oxidation of organic compounds and the genes for the c-type cytochromes required for extracellular electron transfer. Conclusion The results suggest that Pelobacter species evolved parallel strategies to enhance their ability to compete in environments in which electron acceptors for anaerobic respiration were limiting. More generally, these results demonstrate how relatively few gene changes can dramatically transform metabolic capabilities and expand the range of environments in which microorganisms can compete.

  16. Electricity Recovery from Municipal Sewage Wastewater Using a Hydrogel Complex Composed of Microbially Reduced Graphene Oxide and Sludge

    Directory of Open Access Journals (Sweden)

    Naoko Yoshida

    2016-08-01

    Full Text Available Graphene oxide (GO has recently been shown to be an excellent anode substrate for exoelectrogens. This study demonstrates the applicability of GO in recovering electricity from sewage wastewater. Anaerobic incubation of sludge with GO formed a hydrogel complex that embeds microbial cells via π-π stacking of microbially reduced GO. The rGO complex was electrically conductive (23 mS·cm−1 and immediately produced electricity in sewage wastewater under polarization at +200 mV vs. Ag/AgCl. Higher and more stable production of electricity was observed with rGO complexes (179–310 μA·cm−3 than with graphite felt (GF; 79–95 μA·cm−3. Electrochemical analyses revealed that this finding was attributable to the greater capacitance and smaller internal resistance of the rGO complex. Microbial community analysis showed abundances of Geobacter species in both rGO and GF complexes, whereas more diverse candidate exoelectrogens in the Desulfarculaceae family and Geothrix genus were particularly prominent in the rGO complex.

  17. A biofilm microreactor system for simultaneous electrochemical and nuclear magnetic resonance techniques

    International Nuclear Information System (INIS)

    Renslow, Ryan S.; Babauta, Jerome T.; Majors, Paul D.; Mehta, Hardeep S.; Ewing, R. James; Ewing, Thomas; Mueller, Karl T.; Beyenal, Haluk

    2014-01-01

    In order to fully understand electrochemically active biofilms and the limitations to their scale-up in industrial biofilm reactors, a complete picture of the microenvironments inside the biofilm is needed. Nuclear magnetic resonance (NMR) techniques are ideally suited for the study of biofilms and for probing their microenvironments because these techniques allow for non-invasive interrogation and in situ monitoring with high resolution. By combining NMR with simultaneous electrochemical techniques, it is possible to sustain and study live electrochemically active biofilms. Here, we introduce a novel biofilm microreactor system that allows for simultaneous electrochemical and NMR techniques (EC-NMR) at the microscale. Microreactors were designed with custom radiofrequency resonator coils, which allowed for NMR measurements of biofilms growing on polarized gold electrodes. For an example application of this system, we grew Geobacter sulfurreducens biofilms. NMR was used to investigate growth media flow velocities, which were compared to simulated laminar flow, and electron donor concentrations inside the biofilms. We use Monte Carlo error analysis to estimate standard deviations of the electron donor concentration measurements within the biofilm. The EC-NMR biofilm microreactor system can ultimately be used to correlate extracellular electron transfer rates with metabolic reactions and explore extracellular electron transfer mechanisms

  18. New insights into enhanced anaerobic degradation of coal gasification wastewater (CGW) with the assistance of graphene.

    Science.gov (United States)

    Zhu, Hao; Han, Yuxing; Ma, Wencheng; Han, Hongjun; Ma, Weiwei; Xu, Chunyan

    2018-08-01

    The up-flow anaerobic sludge blanket (UASB) system with graphene assisted was developed for coal gasification wastewater (CGW) treatment. Short-term results showed that optimal graphene addition (0.5 g/L) resulted in a more significant enhancement of methane production and chemical oxygen demand (COD) removal compared with that of the optimal activated carbon addition (10.0 g/L). Long-term results demonstrated that COD removal efficiency and methane production rate with graphene assisted achieved 64.7% and 180.5 mL/d, respectively. In addition, graphene could promote microbes accumulation and enzymes activity, resulting in higher extracellular polymeric substances (EPS) and coenzyme F 420 concentrations. X-ray Diffraction (XRD) analysis indicated that chemical of graphene changed insignificantly during the experiment. Meanwhile, with graphene assisted, cells were attached together to form microbial aggregates to facilitate sludge granulation process. Furthermore, the enriched Geobacter and Pseudomonas might perform direct interspecies electron transfer (DIET) with Methanosaeta via biological electrical connection, enhancing the anaerobic degradation of CGW. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. Uranium fate in wetland mesocosms: Effects of plants at two ...

    Science.gov (United States)

    Small-scale continuous flow wetland mesocosms (~0.8 L) were used to evaluate how plant roots under different iron loadings affect uranium (U) mobility. When significant concentrations of ferrous iron (Fe) were present at circumneutral pH values, U concentrations in root exposed sediments were an order of magnitude greater than concentrations in root excluded sediments. Micro X-ray absorption near-edge structure (µ-XANES) spectroscopy indicated that U was associated with the plant roots primarily as U(VI) or U(V), with limited evidence of U(IV). Micro X-ray fluorescence (µ-XRF) of plant roots suggested that for high iron loading at circumneutral pH, U was co-located with Fe, perhaps co-precipitated with root Fe plaques, while for low iron loading at a pH of ~4 the correlation between U and Fe was not significant, consistent with previous observations of U associated with organic matter. Quantitative PCR analyses indicated that the root exposed sediments also contained elevated numbers of Geobacter spp., which are likely associated with enhanced iron cycling, but may also reduce mobile U(VI) to less mobile U(IV) species. There are significant uncertainties regarding the environmental fate of uranium (U) and efforts to minimize U exposures require understanding of its mobility in environmental systems. Much research has focused on sequestering U as solids within groundwater aquifers, where localized risks can be controlled.1 Subsurface sequestration limits t

  20. Uranium Biominerals Precipitated by an Environmental Isolate of Serratia under Anaerobic Conditions

    Science.gov (United States)

    Newsome, Laura; Morris, Katherine; Lloyd, Jonathan. R.

    2015-01-01

    Stimulating the microbially-mediated precipitation of uranium biominerals may be used to treat groundwater contamination at nuclear sites. The majority of studies to date have focussed on the reductive precipitation of uranium as U(IV) by U(VI)- and Fe(III)-reducing bacteria such as Geobacter and Shewanella species, although other mechanisms of uranium removal from solution can occur, including the precipitation of uranyl phosphates via bacterial phosphatase activity. Here we present the results of uranium biomineralisation experiments using an isolate of Serratia obtained from a sediment sample representative of the Sellafield nuclear site, UK. When supplied with glycerol phosphate, this Serratia strain was able to precipitate 1 mM of soluble U(VI) as uranyl phosphate minerals from the autunite group, under anaerobic and fermentative conditions. Under phosphate-limited anaerobic conditions and with glycerol as the electron donor, non-growing Serratia cells could precipitate 0.5 mM of uranium supplied as soluble U(VI), via reduction to nano-crystalline U(IV) uraninite. Some evidence for the reduction of solid phase uranyl(VI) phosphate was also observed. This study highlights the potential for Serratia and related species to play a role in the bioremediation of uranium contamination, via a range of different metabolic pathways, dependent on culturing or in situ conditions. PMID:26132209

  1. Influence of Uranium on Bacterial Communities: A Comparison of Natural Uranium-Rich Soils with Controls

    Science.gov (United States)

    Mondani, Laure; Benzerara, Karim; Carrière, Marie; Christen, Richard; Mamindy-Pajany, Yannick; Février, Laureline; Marmier, Nicolas; Achouak, Wafa; Nardoux, Pascal; Berthomieu, Catherine; Chapon, Virginie

    2011-01-01

    This study investigated the influence of uranium on the indigenous bacterial community structure in natural soils with high uranium content. Radioactive soil samples exhibiting 0.26% - 25.5% U in mass were analyzed and compared with nearby control soils containing trace uranium. EXAFS and XRD analyses of soils revealed the presence of U(VI) and uranium-phosphate mineral phases, identified as sabugalite and meta-autunite. A comparative analysis of bacterial community fingerprints using denaturing gradient gel electrophoresis (DGGE) revealed the presence of a complex population in both control and uranium-rich samples. However, bacterial communities inhabiting uraniferous soils exhibited specific fingerprints that were remarkably stable over time, in contrast to populations from nearby control samples. Representatives of Acidobacteria, Proteobacteria, and seven others phyla were detected in DGGE bands specific to uraniferous samples. In particular, sequences related to iron-reducing bacteria such as Geobacter and Geothrix were identified concomitantly with iron-oxidizing species such as Gallionella and Sideroxydans. All together, our results demonstrate that uranium exerts a permanent high pressure on soil bacterial communities and suggest the existence of a uranium redox cycle mediated by bacteria in the soil. PMID:21998695

  2. Microbial Communities in Contaminated Sediments, Associated with Bioremediation of Uranium to Submicromolar Levels▿

    Science.gov (United States)

    Cardenas, Erick; Wu, Wei-Min; Leigh, Mary Beth; Carley, Jack; Carroll, Sue; Gentry, Terry; Luo, Jian; Watson, David; Gu, Baohua; Ginder-Vogel, Matthew; Kitanidis, Peter K.; Jardine, Philip M.; Zhou, Jizhong; Criddle, Craig S.; Marsh, Terence L.; Tiedje, James M.

    2008-01-01

    Microbial enumeration, 16S rRNA gene clone libraries, and chemical analysis were used to evaluate the in situ biological reduction and immobilization of uranium(VI) in a long-term experiment (more than 2 years) conducted at a highly uranium-contaminated site (up to 60 mg/liter and 800 mg/kg solids) of the U.S. Department of Energy in Oak Ridge, TN. Bioreduction was achieved by conditioning groundwater above ground and then stimulating growth of denitrifying, Fe(III)-reducing, and sulfate-reducing bacteria in situ through weekly injection of ethanol into the subsurface. After nearly 2 years of intermittent injection of ethanol, aqueous U levels fell below the U.S. Environmental Protection Agency maximum contaminant level for drinking water and groundwater (reducers were detected, including Desulfovibrio, Geobacter, Anaeromyxobacter, Desulfosporosinus, and Acidovorax spp. The predominant sulfate-reducing bacterial species were Desulfovibrio spp., while the iron reducers were represented by Ferribacterium spp. and Geothrix spp. Diversity-based clustering revealed differences between treated and untreated zones and also within samples of the treated area. Spatial differences in community structure within the treatment zone were likely related to the hydraulic pathway and to electron donor metabolism during biostimulation. PMID:18456853

  3. Survey of Microbial Diversity in Flood Areas during Thailand 2011 Flood Crisis Using High-Throughput Tagged Amplicon Pyrosequencing.

    Science.gov (United States)

    Mhuantong, Wuttichai; Wongwilaiwalin, Sarunyou; Laothanachareon, Thanaporn; Eurwilaichitr, Lily; Tangphatsornruang, Sithichoke; Boonchayaanant, Benjaporn; Limpiyakorn, Tawan; Pattaragulwanit, Kobchai; Punmatharith, Thantip; McEvoy, John; Khan, Eakalak; Rachakornkij, Manaskorn; Champreda, Verawat

    2015-01-01

    The Thailand flood crisis in 2011 was one of the largest recorded floods in modern history, causing enormous damage to the economy and ecological habitats of the country. In this study, bacterial and fungal diversity in sediments and waters collected from ten flood areas in Bangkok and its suburbs, covering residential and agricultural areas, were analyzed using high-throughput 454 pyrosequencing of 16S rRNA gene and internal transcribed spacer sequences. Analysis of microbial community showed differences in taxa distribution in water and sediment with variations in the diversity of saprophytic microbes and sulfate/nitrate reducers among sampling locations, suggesting differences in microbial activity in the habitats. Overall, Proteobacteria represented a major bacterial group in waters, while this group co-existed with Firmicutes, Bacteroidetes, and Actinobacteria in sediments. Anaeromyxobacter, Steroidobacter, and Geobacter were the dominant bacterial genera in sediments, while Sulfuricurvum, Thiovirga, and Hydrogenophaga predominated in waters. For fungi in sediments, Ascomycota, Glomeromycota, and Basidiomycota, particularly in genera Philipsia, Rozella, and Acaulospora, were most frequently detected. Chytridiomycota and Ascomycota were the major fungal phyla, and Rhizophlyctis and Mortierella were the most frequently detected fungal genera in water. Diversity of sulfate-reducing bacteria, related to odor problems, was further investigated using analysis of the dsrB gene which indicated the presence of sulfate-reducing bacteria of families Desulfobacteraceae, Desulfobulbaceae, Syntrobacteraceae, and Desulfoarculaceae in the flood sediments. The work provides an insight into the diversity and function of microbes related to biological processes in flood areas.

  4. Microbial interspecies electron transfer via electric currents through conductive minerals

    Science.gov (United States)

    Kato, Souichiro; Hashimoto, Kazuhito; Watanabe, Kazuya

    2012-01-01

    In anaerobic biota, reducing equivalents (electrons) are transferred between different species of microbes [interspecies electron transfer (IET)], establishing the basis of cooperative behaviors and community functions. IET mechanisms described so far are based on diffusion of redox chemical species and/or direct contact in cell aggregates. Here, we show another possibility that IET also occurs via electric currents through natural conductive minerals. Our investigation revealed that electrically conductive magnetite nanoparticles facilitated IET from Geobacter sulfurreducens to Thiobacillus denitrificans, accomplishing acetate oxidation coupled to nitrate reduction. This two-species cooperative catabolism also occurred, albeit one order of magnitude slower, in the presence of Fe ions that worked as diffusive redox species. Semiconductive and insulating iron-oxide nanoparticles did not accelerate the cooperative catabolism. Our results suggest that microbes use conductive mineral particles as conduits of electrons, resulting in efficient IET and cooperative catabolism. Furthermore, such natural mineral conduits are considered to provide ecological advantages for users, because their investments in IET can be reduced. Given that conductive minerals are ubiquitously and abundantly present in nature, electric interactions between microbes and conductive minerals may contribute greatly to the coupling of biogeochemical reactions. PMID:22665802

  5. A monetary comparison of energy recovered from microbial fuel cells and microbial electrolysis cells fed winery or domestic wastewaters

    Energy Technology Data Exchange (ETDEWEB)

    Cusick, Roland D.; Kiely, Patrick D.; Logan, Bruce E. [Department of Civil and Environmental Engineering, H2E Center, Penn State University, University Park, PA 16802 (United States)

    2010-09-15

    Microbial fuel (MFCs) and electrolysis cells (MECs) can be used to recover energy directly as electricity or hydrogen from organic matter. Organic removal efficiencies and values of the different energy products were compared for MFCs and MECs fed winery or domestic wastewater. TCOD removal (%) and energy recoveries (kWh/kg-COD) were higher for MFCs than MECs with both wastewaters. At a cost of 4.51/kg-H{sub 2} for winery wastewater and 3.01/kg-H{sub 2} for domestic wastewater, the hydrogen produced using MECs cost less than the estimated merchant value of hydrogen (6/kg-H{sub 2}). 16S rRNA clone libraries indicated the predominance of Geobacter species in anodic microbial communities in MECs for both wastewaters, suggesting low current densities were the result of substrate limitations. The results of this study show that energy recovery and organic removal from wastewater are more effective with MFCs than MECs, but that hydrogen production from wastewater fed MECs can be cost effective. (author)

  6. Current generation in microbial electrolysis cells with addition of amorphous ferric hydroxide, Tween 80, or DNA

    KAUST Repository

    Ren, Lijiao

    2012-11-01

    Iron-oxide nanoparticles and the Tween 80 have previously been shown to improve power generation in microbial fuel cells (MFCs), presumably by improving electron transfer from the bacteria to the anode. We examined whether several chemicals would affect current production in single-chamber microbial electrolysis cells (MECs), where hydrogen gas is produced at the cathode, using mixed cultures and Geobacter sulfurreducens. Tween 80 did not increase the current. Fe(OH) 3 addition increased the maximum current density of both the mixed cultures (from 6.1 ± 0.9 A/m 2 to 8.8 ± 0.3 A/m 2) and pure cultures (from 4.8 ± 0.5 A/m 2 to 7.4 ± 1.1 A/m 2). Improved current production was sustained even after iron was no longer added to the medium. It was demonstrated that increased current resulted from improved cathode performance. Analysis using electrochemical impedance spectroscopy (EIS) showed that the iron primarily reduced the diffusion resistances of the cathodes, and scanning electron microscopy (SEM) images showed the formation of highly porous structures on the cathode. The addition of DNA also did not improve MEC or MFC performance. These results demonstrated that among these treatments only Fe(OH) 3 addition was a viable method for enhancing current densities in MECs, primarily by improving cathode performance. Copyright © 2012, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights.

  7. Operational, design and microbial aspects related to power production with microbial fuel cells implemented in constructed wetlands.

    Science.gov (United States)

    Corbella, Clara; Guivernau, Miriam; Viñas, Marc; Puigagut, Jaume

    2015-11-01

    This work aimed at determining the amount of energy that can be harvested by implementing microbial fuel cells (MFC) in horizontal subsurface constructed wetlands (HSSF CWs) during the treatment of real domestic wastewater. To this aim, MFC were implemented in a pilot plant based on two HSSF CW, one fed with primary settled wastewater (Settler line) and the other fed with the effluent of a hydrolytic up-flow sludge blanket reactor (HUSB line). The eubacterial and archaeal community was profiled on wetland gravel, MFC electrodes and primary treated wastewater by means of 16S rRNA gene-based 454-pyrosequencing and qPCR of 16S rRNA and mcrA genes. Maximum current (219 mA/m(2)) and power (36 mW/m(2)) densities were obtained for the HUSB line. Power production pattern correlated well with water level fluctuations within the wetlands, whereas the type of primary treatment implemented had a significant impact on the diversity and relative abundance of eubacteria communities colonizing MFC. It is worth noticing the high predominance (13-16% of relative abundance) of one OTU belonging to Geobacter on active MFC of the HUSB line that was absent for the settler line MFC. Hence, MFC show promise for power production in constructed wetlands receiving the effluent of a HUSB reactor. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Metal-like transport in proteins: A new paradigm for biological electron transfer

    Science.gov (United States)

    Malvankar, Nikhil; Vargas, Madeline; Tuominen, Mark; Lovley, Derek

    2012-02-01

    Electron flow in biologically proteins generally occurs via tunneling or hopping and the possibility of electron delocalization has long been discounted. Here we report metal-like transport in protein nanofilaments, pili, of bacteria Geobacter sulfurreducens that challenges this long-standing belief [1]. Pili exhibit conductivities comparable to synthetic organic metallic nanostructures. The temperature, magnetic field and gate-voltage dependence of pili conductivity is akin to that of quasi-1D disordered metals, suggesting a metal-insulator transition. Magnetoresistance (MR) data provide evidence for quantum interference and weak localization at room temperature, as well as a temperature and field-induced crossover from negative to positive MR. Furthermore, pili can be doped with protons. Structural studies suggest the possibility of molecular pi stacking in pili, causing electron delocalization. Reducing the disorder increases the metallic nature of pili. These electronically functional proteins are a new class of electrically conductive biological proteins that can be used to generate future generation of inexpensive and environmentally-sustainable nanomaterials and nanolectronic devices such as transistors and supercapacitors. [1] Malvankar et al. Nature Nanotechnology, 6, 573-579 (2011)

  9. Effects of modified biochar on rhizosphere microecology of rice (Oryza sativa L.) grown in As-contaminated soil.

    Science.gov (United States)

    Liu, Shusi; Lu, Yixin; Yang, Chen; Liu, Chuanping; Ma, Lin; Dang, Zhi

    2017-10-01

    Biochar was carbon-rich and generated by high-temperature pyrolysis of biomass under oxygen-limited conditions. Due to the limitations of surface functional groups and the weakness of surface activity in the field of environmental remediation, the raw biochar frequently was chemically modified to improve its properties with a new performance. In this study, a kind of high-efficiency and low-cost amino biochar modified by nano zero-valent iron (ABC/NZVI) was synthesized and applied to paddy soil contaminated with arsenic (As). Dynamic changes of soil properties, arsenic speciations and rhizosphere microbial communities have been investigated over the whole growth period of rice plants. Pot experiments revealed that the ABC/NZVI could decrease the arsenic concentration in rice straw by 47.9% and increase the content of nitrogen in rice straw by 47.2%. Proportion of Geobacter in soil with ABC/NZVI treatment increased by 175% in tillering period; while Nitrososphaera decreased by 61 and 20% in tillering and maturity, respectively, compared to that of control. ABC/NZVI promotes arsenic immobilization in rhizosphere soil and precipitation on root surface and reduces arsenic accumulation in rice. At the same time, ABC/NZVI would inhibit Nitrososphaera which is related to ammonia oxidation process, and it would have a promising potential as soil amendment to reduce nitrogen loss probably.

  10. Stimulation of electro-fermentation in single-chamber microbial electrolysis cells driven by genetically engineered anode biofilms

    Science.gov (United States)

    Awate, Bhushan; Steidl, Rebecca J.; Hamlischer, Thilo; Reguera, Gemma

    2017-07-01

    Unwanted metabolites produced during fermentations reduce titers and productivity and increase the cost of downstream purification of the targeted product. As a result, the economic feasibility of otherwise attractive fermentations is low. Using ethanol fermentation by the consolidated bioprocessing cellulolytic bacterium Cellulomonas uda, we demonstrate the effectiveness of anodic electro-fermentations at maximizing titers and productivity in a single-chamber microbial electrolysis cell (SCMEC) without the need for metabolic engineering of the fermentative microbe. The performance of the SCMEC platform relied on the genetic improvements of anode biofilms of the exoelectrogen Geobacter sulfurreducens that prevented the oxidation of cathodic hydrogen and improved lactate oxidation. Furthermore, a hybrid bioanode was designed that maximized the removal of organic acids in the fermentation broth. The targeted approach increased cellobiose consumption rates and ethanol titers, yields, and productivity three-fold or more, prevented pH imbalances and reduced batch-to-batch variability. In addition, the sugar substrate was fully consumed and ethanol was enriched in the broth during the electro-fermentation, simplifying its downstream purification. Such improvements and the possibility of scaling up SCMEC configurations highlight the potential of anodic electro-fermentations to stimulate fermentative bacteria beyond their natural capacity and to levels required for industrial implementation.

  11. Bacterial community composition in the water column of a lake formed by a former uranium open pit mine.

    Science.gov (United States)

    Edberg, Frida; Andersson, Anders F; Holmström, Sara J M

    2012-11-01

    Mining of pyrite minerals is a major environmental issue involving both biological and geochemical processes. Here we present a study of an artificial lake of a former uranium open pit mine with the aim to connect the chemistry and bacterial community composition (454-pyrosequencing of 16S rRNA genes) in the stratified water column. A shift in the water chemistry from oxic conditions in the epilimnion to anoxic, alkaline, and metal and sulfide-rich conditions in the hypolimnion was corresponded by a strong shift in the bacterial community, with few shared operational taxonomic units (OTU) between the water layers. The epilimnetic bacterial community of the lake (~20 years old) showed similarities to other temperate freshwater lakes, while the hypolimnetic bacterial community showed similarity to extreme chemical environments. The epilimnetic bacterial community had dominance of Actinobacteria and Betaproteobacteria. The hypolimnion displayed a higher bacterial diversity and was dominated by the phototrophic green sulphur bacterium of the genus Chlorobium (ca. 40 % of the total community). Deltaproteobacteria were only represented in the hypolimnion and the most abundant OTUs were affiliated with ferric iron and sulfate reducers of the genus Geobacter and Desulfobulbus, respectively. The chemistry is clearly controlling, especially the hypolimnetic, bacterial community but the community composition also indicates that the bacteria are involved in metal cycling in the lake.

  12. Microbial community composition is unaffected by anode potential

    KAUST Repository

    Zhu, Xiuping

    2014-01-21

    There is great controversy on how different set anode potentials affect the performance of a bioelectrochemical system (BES). It is often reported that more positive potentials improve acclimation and performance of exoelectrogenic biofilms, and alter microbial community structure, while in other studies relatively more negative potentials were needed to achieve higher current densities. To address this issue, the biomass, electroactivity, and community structure of anodic biofilms were examined over a wide range of set anode potentials (-0.25, -0.09, 0.21, 0.51, and 0.81 V vs a standard hydrogen electrode, SHE) in single-chamber microbial electrolysis cells. Maximum currents produced using a wastewater inoculum increased with anode potentials in the range of -0.25 to 0.21 V, but decreased at 0.51 and 0.81 V. The maximum currents were positively correlated with increasing biofilm biomass. Pyrosequencing indicated biofilm communities were all similar and dominated by bacteria most similar to Geobacter sulfurreducens. Differences in anode performance with various set potentials suggest that the exoelectrogenic communities self-regulate their exocellular electron transfer pathways to adapt to different anode potentials. © 2013 American Chemical Society.

  13. Progressive biogeochemical transformation of placer gold particles drives compositional changes in associated biofilm communities.

    Science.gov (United States)

    Rea, Maria Angelica; Standish, Christopher D; Shuster, Jeremiah; Bissett, Andrew; Reith, Frank

    2018-05-03

    Biofilms on placer gold (Au)-particle surfaces drive Au solubilization and re-concentration thereby progressively transforming the particles. Gold solubilization induces Au-toxicity; however, Au-detoxifying community members ameliorates Au-toxicity by precipitating soluble Au to metallic Au. We hypothesize that Au-dissolution and re-concentration (precipitation) places selective pressures on associated microbial communities, leading to compositional changes and subsequent Au-particle transformation. We analyzed Au-particles from eight United Kingdom sites using next generation sequencing, electron microscopy and micro-analyses. Gold particles contained biofilms composed of prokaryotic cells and extracellular polymeric substances intermixed with (bio)minerals. Across all sites communities were dominated by Proteobacteria (689, 97% Operational Taxonomic Units, 59.3% of total reads), with β-Proteobacteria being the most abundant. A wide range of Au-morphotypes including nanoparticles, micro-crystals, sheet-like Au and secondary rims, indicated that dissolution and re-precipitation occurred, and from this transformation indices were calculated. Multivariate statistical analyses showed a significant relationship between the extent of Au-particle transformation and biofilm community composition, with putative metal-resistant Au-cycling taxa linked to progressive Au transformation. These included the genera Pseudomonas, Leptothrix and Acinetobacter. Additionally, putative exoelectrogenic genera Rhodoferax and Geobacter were highly abundant. In conclusion, biogeochemical Au-cycling and Au-particle transformation occurred at all sites and exerted a strong influence on biofilm community composition.

  14. Impact of biostimulated redox processes on metal dynamics in an iron-rich creek soil of a former uranium mining area.

    Science.gov (United States)

    Burkhardt, Eva-Maria; Akob, Denise M; Bischoff, Sebastian; Sitte, Jana; Kostka, Joel E; Banerjee, Dipanjan; Scheinost, Andreas C; Küsel, Kirsten

    2010-01-01

    Understanding the dynamics of metals and radionuclides in soil environments is necessary for evaluating risks to pristine sites. An iron-rich creek soil of a former uranium-mining district (Ronneburg, Germany) showed high porewater concentrations of heavy metals and radionuclides. Thus, this study aims to (i) evaluate metal dynamics during terminal electron accepting processes (TEAPs) and (ii) characterize active microbial populations in biostimulated soil microcosms using a stable isotope probing (SIP) approach. In biostimulated soil slurries, concentrations of soluble Co, Ni, Zn, As, and unexpectedly U increased during Fe(III)-reduction. This suggests that there was a release of sorbed metals and As during reductive dissolution of Fe(III)-oxides. Subsequent sulfate-reduction was concurrent with a decrease of U, Co, Ni, and Zn concentrations. The relative contribution of U(IV) in the solid phase changed from 18.5 to 88.7% after incubation. The active Fe(III)-reducing population was dominated by delta-Proteobacteria (Geobacter) in (13)C-ethanol amended microcosms. A more diverse community was present in (13)C-lactate amended microcosms including taxa related to Acidobacteria, Firmicutes, delta-Proteobacteria, and beta-Proteobacteria. Our results suggested that biostimulated Fe(III)-reducing communities facilitated the release of metals including U to groundwater which is in contrast to other studies.

  15. Repression of hydrogen uptake using conjugated oligoelectrolytes in microbial electrolysis cells

    KAUST Repository

    Hou, Huijie; Chen, Xiaofen; Liu, Jia; Zhu, Xiuping; Bazan, Guillermo C.; Logan, Bruce E.

    2014-01-01

    Copyright © 2014, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved. DSBN+, a conjugated oligoelectrolyte (COE), was added to microbial electrolysis cells (MECs) to improve hydrogen recovery. The volume of hydrogen gas recovered in a fedbatch cycle of mixed culture MECs increased by 126× compared to controls (no COE addition), mainly by preventing the loss of hydrogen to methane production. Performance in pure culture MECs fed with Geobacter sulfurreducens increased by factors of 10.5 in terms of energy yield, 2.1 in COD removal, and 11.8 in hydrogen yield. Hydrogen gas recycling was reduced, and the volume of hydrogen gas recovered increased by 6.5× compared to controls. Minimal methane production and a lack of hydrogen gas uptake by G. sulfurreducens suggested that the COEs increased hydrogen recoveries by interfering with hydrogen uptake by hydrogenotrophic methanogens but also by exoelectrogenic bacteria. COEs may therefore be useful for inhibiting the activities of certain hydrogenases, although the mechanism of inhibition needs further investigation.

  16. Viable cold-tolerant iron-reducing microorganisms in geographically diverse subglacial environments

    Science.gov (United States)

    Nixon, Sophie L.; Telling, Jon P.; Wadham, Jemma L.; Cockell, Charles S.

    2017-03-01

    Subglacial environments are known to harbour metabolically diverse microbial communities. These microbial communities drive chemical weathering of underlying bedrock and influence the geochemistry of glacial meltwater. Despite its importance in weathering reactions, the microbial cycling of iron in subglacial environments, in particular the role of microbial iron reduction, is poorly understood. In this study we address the prevalence of viable iron-reducing microorganisms in subglacial sediments from five geographically isolated glaciers. Iron-reducing enrichment cultures were established with sediment from beneath Engabreen (Norway), Finsterwalderbreen (Svalbard), Leverett and Russell glaciers (Greenland), and Lower Wright Glacier (Antarctica). Rates of iron reduction were higher at 4 °C compared with 15 °C in all but one duplicated second-generation enrichment culture, indicative of cold-tolerant and perhaps cold-adapted iron reducers. Analysis of bacterial 16S rRNA genes indicates Desulfosporosinus were the dominant iron-reducing microorganisms in low-temperature Engabreen, Finsterwalderbreen and Lower Wright Glacier enrichments, and Geobacter dominated in Russell and Leverett enrichments. Results from this study suggest microbial iron reduction is widespread in subglacial environments and may have important implications for global biogeochemical iron cycling and export to marine ecosystems.

  17. Enhanced treatment of Fischer-Tropsch wastewater using up-flow anaerobic sludge blanket system coupled with micro-electrolysis cell: A pilot scale study.

    Science.gov (United States)

    Wang, Dexin; Han, Yuxing; Han, Hongjun; Li, Kun; Xu, Chunyan

    2017-08-01

    The coupling of micro-electrolysis cell (MEC) with an up-flow anaerobic sludge blanket (UASB) system in pilot scale was established for enhanced treatment of Fischer-Tropsch (F-T) wastewater. The lowest influent pH (4.99±0.10) and reduced alkali addition were accomplished under the assistance of anaerobic effluent recycling of 200% (stage 5). Simultaneously, the optimum COD removal efficiency (93.5±1.6%) and methane production (2.01±0.13m 3 /m 3 ·d) at the lower hydraulic retention time (HRT) were achieved in this stage. In addition, the dissolved iron from MEC could significantly increase the protein content of tightly bound extracellular polymeric substances (TB-EPS), which was beneficial to formation of stable granules. Furthermore, the high-throughput 16S rRNA gene pyrosequencing in this study further confirmed that Geobacter species could utilize iron oxides particles as electron conduit to perform the direct interspecies electron transfer (DIET) with Methanothrix, finally facilitating the syntrophic degradation of propionic acid and butyric acid and contributing completely methane production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Microbial community analysis in rice paddy soils irrigated by acid mine drainage contaminated water.

    Science.gov (United States)

    Sun, Min; Xiao, Tangfu; Ning, Zengping; Xiao, Enzong; Sun, Weimin

    2015-03-01

    Five rice paddy soils located in southwest China were selected for geochemical and microbial community analysis. These rice fields were irrigated with river water which was contaminated by Fe-S-rich acid mine drainage. Microbial communities were characterized by high-throughput sequencing, which showed 39 different phyla/groups in these samples. Among these phyla/groups, Proteobacteria was the most abundant phylum in all samples. Chloroflexi, Acidobacteria, Nitrospirae, and Bacteroidetes exhibited higher relative abundances than other phyla. A number of rare and candidate phyla were also detected. Moreover, canonical correspondence analysis suggested that pH, sulfate, and nitrate were significant factors that shaped the microbial community structure. In addition, a wide diversity of Fe- and S-related bacteria, such as GOUTA19, Shewanella, Geobacter, Desulfobacca, Thiobacillus, Desulfobacterium, and Anaeromyxobacter, might be responsible for biogeochemical Fe and S cycles in the tested rice paddy soils. Among the dominant genera, GOUTA19 and Shewanella were seldom detected in rice paddy soils.

  19. Hydrogen production with nickel powder cathode catalysts in microbial electrolysis cells

    KAUST Repository

    Selembo, Priscilla A.

    2010-01-01

    Although platinum is commonly used as catalyst on the cathode in microbial electrolysis cells (MEC), non-precious metal alternatives are needed to reduce costs. Cathodes were constructed using a nickel powder (0.5-1 μm) and their performance was compared to conventional electrodes containing Pt (0.002 μm) in MECs and electrochemical tests. The MEC performance in terms of coulombic efficiency, cathodic, hydrogen and energy recoveries were similar using Ni or Pt cathodes, although the maximum hydrogen production rate (Q) was slightly lower for Ni (Q = 1.2-1.3 m3 H2/m3/d; 0.6 V applied) than Pt (1.6 m3 H2/m3/d). Nickel dissolution was minimized by replacing medium in the reactor under anoxic conditions. The stability of the Ni particles was confirmed by examining the cathodes after 12 MEC cycles using scanning electron microscopy and linear sweep voltammetry. Analysis of the anodic communities in these reactors revealed dominant populations of Geobacter sulfurreduces and Pelobacter propionicus. These results demonstrate that nickel powder can be used as a viable alternative to Pt in MECs, allowing large scale production of cathodes with similar performance to systems that use precious metal catalysts. © 2009 Professor T. Nejat Veziroglu.

  20. Repression of hydrogen uptake using conjugated oligoelectrolytes in microbial electrolysis cells

    KAUST Repository

    Hou, Huijie

    2014-11-01

    Copyright © 2014, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved. DSBN+, a conjugated oligoelectrolyte (COE), was added to microbial electrolysis cells (MECs) to improve hydrogen recovery. The volume of hydrogen gas recovered in a fedbatch cycle of mixed culture MECs increased by 126× compared to controls (no COE addition), mainly by preventing the loss of hydrogen to methane production. Performance in pure culture MECs fed with Geobacter sulfurreducens increased by factors of 10.5 in terms of energy yield, 2.1 in COD removal, and 11.8 in hydrogen yield. Hydrogen gas recycling was reduced, and the volume of hydrogen gas recovered increased by 6.5× compared to controls. Minimal methane production and a lack of hydrogen gas uptake by G. sulfurreducens suggested that the COEs increased hydrogen recoveries by interfering with hydrogen uptake by hydrogenotrophic methanogens but also by exoelectrogenic bacteria. COEs may therefore be useful for inhibiting the activities of certain hydrogenases, although the mechanism of inhibition needs further investigation.

  1. Assessment of microbial communities associated with fermentative-methanogenic biodegradation of aromatic hydrocarbons in groundwater contaminated with a biodiesel blend (B20).

    Science.gov (United States)

    Ramos, Débora Toledo; da Silva, Márcio Luís Busi; Nossa, Carlos Wolfgang; Alvarez, Pedro J J; Corseuil, Henry Xavier

    2014-09-01

    A controlled field experiment was conducted to assess the potential for fermentative-methanogenic biostimulation (by ammonium-acetate injection) to enhance biodegradation of benzene, toluene, ethylbenzene and xylenes (BTEX) as well as polycyclic aromatic hydrocarbons (PAHs) in groundwater contaminated with biodiesel B20 (20:80 v/v soybean biodiesel and diesel). Changes in microbial community structure were assessed by pyrosequencing 16S rRNA analyses. BTEX and PAH removal began 0.7 year following the release, concomitantly with the increase in the relative abundance of Desulfitobacterium and Geobacter spp. (from 5 to 52.7 % and 15.8 to 37.3 % of total Bacteria 16S rRNA, respectively), which are known to anaerobically degrade hydrocarbons. The accumulation of anaerobic metabolites acetate and hydrogen that could hinder the thermodynamic feasibility of BTEX and PAH biotransformations under fermentative/methanogenic conditions was apparently alleviated by the growing predominance of Methanosarcina. This suggests the importance of microbial population shifts that enrich microorganisms capable of interacting syntrophically to enhance the feasibility of fermentative-methanogenic bioremediation of biodiesel blend releases.

  2. Cascade degradation of organic matters in brewery wastewater using a continuous stirred microbial electrochemical reactor and analysis of microbial communities

    Science.gov (United States)

    Wang, Haiman; Qu, Youpeng; Li, Da; Ambuchi, John J.; He, Weihua; Zhou, Xiangtong; Liu, Jia; Feng, Yujie

    2016-01-01

    A continuous stirred microbial electrochemical reactor (CSMER), comprising of a complete mixing zone (CMZ) and microbial electrochemical zone (MEZ), was used for brewery wastewater treatment. The system realized 75.4 ± 5.7% of TCOD and 64.9 ± 4.9% of TSS when fed with brewery wastewater concomitantly achieving an average maximum power density of 304 ± 31 m W m−2. Cascade utilization of organic matters made the CSMER remove a wider range of substrates compared with a continuous stirred tank reactor (CSTR), in which process 79.1 ± 5.6% of soluble protein and 86.6 ± 2.2% of soluble carbohydrates were degraded by anaerobic digestion in the CMZ and short-chain volatile fatty acids were further decomposed and generated current in the MEZ. Co-existence of fermentative bacteria (Clostridium and Bacteroides, 19.7% and 5.0%), acetogenic bacteria (Syntrophobacter, 20.8%), methanogenic archaea (Methanosaeta and Methanobacterium, 40.3% and 38.4%) and exoelectrogens (Geobacter, 12.4%) as well as a clear spatial distribution and syntrophic interaction among them contributed to the cascade degradation process in CSMER. The CSMER shows great promise for practical wastewater treatment application due to high pre-hydrolysis and acidification rate, high energy recovery and low capital cost. PMID:27270788

  3. Change in microbial communities in acetate- and glucose-fed microbial fuel cells in the presence of light

    KAUST Repository

    Xing, Defeng

    2009-09-01

    Power densities produced by microbial fuel cells (MFCs) in natural systems are changed by exposure to light through the enrichment of photosynthetic microorganisms. When MFCs with brush anodes were exposed to light (4000 lx), power densities increased by 8-10% for glucose-fed reactors, and 34% for acetate-fed reactors. Denaturing gradient gel electrophoresis (DGGE) profiles based on the 16S rRNA gene showed that exposure to high light levels changed the microbial communities on the anodes. Based on 16S rRNA gene clone libraries of light-exposed systems the anode communities using glucose were also significantly different than those fed acetate. Dominant bacteria that are known exoelectrogens were identified in the anode biofilm, including a purple nonsulfur (PNS) photosynthetic bacterium, Rhodopseudomonas palustris, and a dissimilatory iron-reducing bacterium, Geobacter sulfurreducens. Pure culture tests confirmed that PNS photosynthetic bacteria increased power production when exposed to high light intensities (4000 lx). These results demonstrate that power production and community composition are affected by light conditions as well as electron donors in single-chamber air-cathode MFCs. © 2009 Elsevier B.V. All rights reserved.

  4. Microbial community composition is unaffected by anode potential

    KAUST Repository

    Zhu, Xiuping; Yates, Matthew D.; Hatzell, Marta C.; Rao, Hari Ananda; Saikaly, Pascal; Logan, Bruce E.

    2014-01-01

    There is great controversy on how different set anode potentials affect the performance of a bioelectrochemical system (BES). It is often reported that more positive potentials improve acclimation and performance of exoelectrogenic biofilms, and alter microbial community structure, while in other studies relatively more negative potentials were needed to achieve higher current densities. To address this issue, the biomass, electroactivity, and community structure of anodic biofilms were examined over a wide range of set anode potentials (-0.25, -0.09, 0.21, 0.51, and 0.81 V vs a standard hydrogen electrode, SHE) in single-chamber microbial electrolysis cells. Maximum currents produced using a wastewater inoculum increased with anode potentials in the range of -0.25 to 0.21 V, but decreased at 0.51 and 0.81 V. The maximum currents were positively correlated with increasing biofilm biomass. Pyrosequencing indicated biofilm communities were all similar and dominated by bacteria most similar to Geobacter sulfurreducens. Differences in anode performance with various set potentials suggest that the exoelectrogenic communities self-regulate their exocellular electron transfer pathways to adapt to different anode potentials. © 2013 American Chemical Society.

  5. Nitrogen loss from anaerobic ammonium oxidation coupled to Iron(III) reduction in a riparian zone.

    Science.gov (United States)

    Ding, Bangjing; Li, Zhengkui; Qin, Yunbin

    2017-12-01

    Anaerobic ammonium oxidation coupled to iron(III) reduction (termed Feammox) is a recently discovered pathway of nitrogen cycling. However, little is known about the pathways of N transformation via Feammox process in riparian zones. In this study, evidence for Feammox in riparian zones with or without vegetation cover was demonstrated using isotope tracing technique and high-throughput sequencing technology. The results showed that Feammox could occur in riparian zones, and demonstrated that N 2 directly from Feammox was dominant Feammox pathway. The Feammox rates in vegetated soil samples was 0.32-0.37 mg N kg -1 d -1 , which is higher than that in un-vegetated soil samples (0.20 mg N kg -1 d -1 ). Moreover, the growth of vegetation led to a 4.99-6.41% increase in the abundance of iron reducing bacteria (Anaeromyxobacter, Pseudomonas and Geobacter) and iron reducing bacteria play an essential role in Feammox process. An estimated loss of 23.7-43.9 kg N ha -1 year -1 was associated with Feammox in the examined riparian zone. Overall, the co-occurrence of ammonium oxidation and iron reduction suggest that Feammox can play an essential role in the pathway of nitrogen removal in riparian zones. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Enhanced phosphorus reduction in simulated eutrophic water: a comparative study of submerged macrophytes, sediment microbial fuel cells, and their combination.

    Science.gov (United States)

    Xu, Peng; Xiao, Enrong; Xu, Dan; Li, Juan; Zhang, Yi; Dai, Zhigang; Zhou, Qiaohong; Wu, Zhenbin

    2018-05-01

    The phosphorus reduction in water column was attempted by integrating sediment microbial fuel cells (SMFCs) with the submerged macrophyte Vallisneria spiralis. A comparative study was conducted to treat simulated water rich in phosphate with a control and three treatments: SMFC alone (SMFC), submerged macrophytes alone (macophyte), and combined macrophytes and fuel cells (M-SMFC). All treatments promoted phosphorus flux from the water column to sediments. Maximum phosphorus reduction was obtained in proportion to the highest stable phosphorus level in sediments in M-SMFC. For the initial phosphate concentrations of 0.2, 1, 2, and 4 mg/L, average phosphate values in the overlying water during four phases decreased by 33.3% (25.0%, 8.3%), 30.8% (5.1%, 17.9%), 36.5% (27.8%, 15.7%), and 36.2% (0.7%, 22.1%) for M-SMFC (macrophyte, SMFC), compared with the control. With macrophyte treatment, the obvious phosphorus release from sediments was observed during the declining period. However, such phenomenon was significantly inhibited with M-SMFC. The electrogenesis bacteria achieved stronger phosphorus adsorption and assimilation was significantly enriched on the closed-circuit anodes. The higher abundance of Geobacter and Pseudomonas in M-SMFC might in part explain the highest phosphorus reduction in the water column. M-SMFC treatment could be promising to control the phosphorus in eutrophic water bodies.

  7. Enrichment of specific electro-active microorganisms and enhancement of methane production by adding granular activated carbon in anaerobic reactors.

    Science.gov (United States)

    Lee, Jung-Yeol; Lee, Sang-Hoon; Park, Hee-Deung

    2016-04-01

    Direct interspecies electron transfer (DIET) via conductive materials can provide significant benefits to anaerobic methane formation in terms of production amount and rate. Although granular activated carbon (GAC) demonstrated its applicability in facilitating DIET in methanogenesis, DIET in continuous flow anaerobic reactors has not been verified. Here, evidences of DIET via GAC were explored. The reactor supplemented with GAC showed 1.8-fold higher methane production rate than that without GAC (35.7 versus 20.1±7.1mL-CH4/d). Around 34% of methane formation was attributed to the biomass attached to GAC. Pyrosequencing of 16S rRNA gene demonstrated the enrichment of exoelectrogens (e.g. Geobacter) and hydrogenotrophic methanogens (e.g. Methanospirillum and Methanolinea) from the biomass attached to GAC. Furthermore, anodic and cathodic currents generation was observed in an electrochemical cell containing GAC biomass. Taken together, GAC supplementation created an environment for enriching the microorganisms involved in DIET, which increased the methane production rate. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Polyphasic analysis of an Azoarcus-Leptothrix-dominated bacterial biofilm developed on stainless steel surface in a gasoline-contaminated hypoxic groundwater.

    Science.gov (United States)

    Benedek, Tibor; Táncsics, András; Szabó, István; Farkas, Milán; Szoboszlay, Sándor; Fábián, Krisztina; Maróti, Gergely; Kriszt, Balázs

    2016-05-01

    Pump and treat systems are widely used for hydrocarbon-contaminated groundwater remediation. Although biofouling (formation of clogging biofilms on pump surfaces) is a common problem in these systems, scarce information is available regarding the phylogenetic and functional complexity of such biofilms. Extensive information about the taxa and species as well as metabolic potential of a bacterial biofilm developed on the stainless steel surface of a pump submerged in a gasoline-contaminated hypoxic groundwater is presented. Results shed light on a complex network of interconnected hydrocarbon-degrading chemoorganotrophic and chemolitotrophic bacteria. It was found that besides the well-known hydrocarbon-degrading aerobic/facultative anaerobic biofilm-forming organisms (e.g., Azoarcus, Leptothrix, Acidovorax, Thauera, Pseudomonas, etc.), representatives of Fe(2+)-and Mn(2+)-oxidizing (Thiobacillus, Sideroxydans, Gallionella, Rhodopseudomonas, etc.) as well as of Fe(3+)- and Mn(4+)-respiring (Rhodoferax, Geobacter, Magnetospirillum, Sulfurimonas, etc.) bacteria were present in the biofilm. The predominance of β-Proteobacteria within the biofilm bacterial community in phylogenetic and functional point of view was revealed. Investigation of meta-cleavage dioxygenase and benzylsuccinate synthase (bssA) genes indicated that within the biofilm, Azoarcus, Leptothrix, Zoogloea, and Thauera species are most probably involved in intrinsic biodegradation of aromatic hydrocarbons. Polyphasic analysis of the biofilm shed light on the fact that subsurface microbial accretions might be reservoirs of novel putatively hydrocarbon-degrading bacterial species. Moreover, clogging biofilms besides their detrimental effects might supplement the efficiency of pump and treat systems.

  9. Electromicrobiology of Dissimilatory Sulfur Reducing Bacterium Desulfuromonas acetexigens

    KAUST Repository

    Bin Bandar, Khaled

    2014-12-01

    Bioelectrochmical systems (BES) are engineered electrochemical devices that harness hidden chemical energy of the wastewater in to the form of electricity or hydrogen. Unique microbial communities enrich in these systems for oxidation of organic matter as well as transfer of resulted electron to anode, known them as “electricigens” communities. Exploring novel electricigenesis microbial communities in the nature and understanding their electromicrobiology is one the important aspect for BES systems scale up. Herein, we report first time the electricigenesis property of an anaerobic, fresh water sediment, sulfur reducing bacterium Desulfuromona acetexigens. The electrochemical behavior of D. acetexigens biofilms grown on graphite-rod electrodes in batch-fed mode under an applied potential was investigated with traditional electroanalytical tools, and correlate the electron transfer from biofilms to electrode with a model electricigen Geobacter sulfurreducens electrochemical behavior. Research findings suggest that D. acetexigens has the ability to use electrode as electron acceptor in BES systems through establishing the direct contact with anode by expressing the membrane bound redox proteins, but not due to the secretion of soluble redox mediators. Preliminary results revealed that D. acetexigens express three distinct redox proteins in their membranes for turnover of the electrons from biofilm to electrode, and the 4 whole electricigenesis process observed to be unique in the D. acetexigens compared to that of well-studied model organism G. sulfurreducens.

  10. Microbial community analysis of switchgrass planted and unplanted soil microcosms displaying PCB dechlorination.

    Science.gov (United States)

    Liang, Yi; Meggo, Richard; Hu, Dingfei; Schnoor, Jerald L; Mattes, Timothy E

    2015-08-01

    Polychlorinated biphenyls (PCBs) pose potential risks to human and environmental health because they are carcinogenic, persistent, and bioaccumulative. In this study, we investigated bacterial communities in soil microcosms spiked with PCB 52, 77, and 153. Switchgrass (Panicum virgatum) was employed to improve overall PCB removal, and redox cycling (i.e., sequential periods of flooding followed by periods of no flooding) was performed in an effort to promote PCB dechlorination. Lesser chlorinated PCB transformation products were detected in all microcosms, indicating the occurrence of PCB dechlorination. Terminal restriction fragment length polymorphism (T-RFLP) and clone library analysis showed that PCB spiking, switchgrass planting, and redox cycling affected the microbial community structure. Putative organohalide-respiring Chloroflexi populations, which were not found in unflooded microcosms, were enriched after 2 weeks of flooding in the redox-cycled microcosms. Sequences classified as Geobacter sp. were detected in all microcosms and were most abundant in the switchgrass-planted microcosm spiked with PCB congeners. The presence of possible organohalide-respiring bacteria in these soil microcosms suggests that they play a role in PCB dechlorination therein.

  11. Active transport, substrate specificity, and methylation of Hg(II) in anaerobic bacteria

    Science.gov (United States)

    Schaefer, Jeffra K.; Rocks, Sara S.; Zheng, Wang; Liang, Liyuan; Gu, Baohua; Morel, François M. M.

    2011-01-01

    The formation of methylmercury (MeHg), which is biomagnified in aquatic food chains and poses a risk to human health, is effected by some iron- and sulfate-reducing bacteria (FeRB and SRB) in anaerobic environments. However, very little is known regarding the mechanism of uptake of inorganic Hg by these organisms, in part because of the inherent difficulty in measuring the intracellular Hg concentration. By using the FeRB Geobacter sulfurreducens and the SRB Desulfovibrio desulfuricans ND132 as model organisms, we demonstrate that Hg(II) uptake occurs by active transport. We also establish that Hg(II) uptake by G. sulfurreducens is highly dependent on the characteristics of the thiols that bind Hg(II) in the external medium, with some thiols promoting uptake and methylation and others inhibiting both. The Hg(II) uptake system of D. desulfuricans has a higher affinity than that of G. sulfurreducens and promotes Hg methylation in the presence of stronger complexing thiols. We observed a tight coupling between Hg methylation and MeHg export from the cell, suggesting that these two processes may serve to avoid the build up and toxicity of cellular Hg. Our results bring up the question of whether cellular Hg uptake is specific for Hg(II) or accidental, occurring via some essential metal importer. Our data also point at Hg(II) complexation by thiols as an important factor controlling Hg methylation in anaerobic environments. PMID:21555571

  12. Photochemical reactions between mercury (Hg) and dissolved organic matter decrease Hg bioavailability and methylation.

    Science.gov (United States)

    Luo, Hong-Wei; Yin, Xiangping; Jubb, Aaron M; Chen, Hongmei; Lu, Xia; Zhang, Weihua; Lin, Hui; Yu, Han-Qing; Liang, Liyuan; Sheng, Guo-Ping; Gu, Baohua

    2017-01-01

    Atmospheric deposition of mercury (Hg) to surface water is one of the dominant sources of Hg in aquatic environments and ultimately drives methylmercury (MeHg) toxin accumulation in fish. It is known that freshly deposited Hg is more readily methylated by microorganisms than aged or preexisting Hg; however the underlying mechanism of this process is unclear. We report that Hg bioavailability is decreased by photochemical reactions between Hg and dissolved organic matter (DOM) in water. Photo-irradiation of Hg-DOM complexes results in loss of Sn(II)-reducible (i.e. reactive) Hg and up to an 80% decrease in MeHg production by the methylating bacterium Geobacter sulfurreducens PCA. Loss of reactive Hg proceeded at a faster rate with a decrease in the Hg to DOM ratio and is attributed to the possible formation of mercury sulfide (HgS). These results suggest a new pathway of abiotic photochemical formation of HgS in surface water and provide a mechanism whereby freshly deposited Hg is readily methylated but, over time, progressively becomes less available for microbial uptake and methylation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Carbon nanotube fiber mats for microbial fuel cell electrodes.

    Science.gov (United States)

    Delord, Brigitte; Neri, Wilfrid; Bertaux, Karen; Derre, Alain; Ly, Isabelle; Mano, Nicolas; Poulin, Philippe

    2017-11-01

    Novel carbon nanotube based electrodes of microbial fuel cells (MFC) have been developed. MFC is a promising technology for the wastewater treatment and the production of electrical energy from redox reactions of natural substrates. Performances of such bio-electrochemical systems depend critically on the structure and properties of the electrodes. The presently developed materials are made by weaving fibers solely comprised of carbon nanotubes. They exhibit a large scale porosity controlled by the weaving process. This porosity allows an easy colonization by electroactive bacteria. In addition, the fibers display a nanostructuration that promotes excellent growth and adhesion of the bacteria at the surface of the electrodes. This unique combination of large scale porosity and nanostructuration allows the present electrodes to perform better than carbon reference. When used as anode in a bioelectrochemical reactor in presence of Geobacter sulfurreducens bacteria, the present electrodes show a maximal current density of about 7.5mA/cm 2 . Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Strong shift in the diazotrophic endophytic bacterial community inhabiting rice (Oryza sativa) plants after flooding.

    Science.gov (United States)

    Ferrando, Lucía; Fernández Scavino, Ana

    2015-09-01

    Flooding impacts soil microbial communities, but its effect on endophytic communities has rarely been explored. This work addresses the effect of flooding on the abundance and diversity of endophytic diazotrophic communities on rice plants established in a greenhouse experiment. The nifH gene was significantly more abundant in roots after flooding, whereas the nifH gene copy numbers in leaves were unaffected and remained low. The PCA (principal component analysis) of T-RFLP (terminal restriction fragment length polymorphism) profiles indicated that root communities of replicate plots were more similar and diverse after flooding than before flooding. The nifH libraries obtained by cloning and 454 pyrosequencing consistently showed a remarkable shift in the diazotrophic community composition after flooding. Gammaproteobacteria (66-98%), mainly of the genus Stenotrophomonas, prevailed in roots before flooding, whereas Betaproteobacteria was the dominant class (26-34%) after flooding. A wide variety of aerotolerant and anaerobic diazotrophic bacteria (e.g. Dechloromonas, Rhodopseudomonas, Desulfovibrio, Geobacter, Chlorobium, Spirochaeta, Selenomonas and Dehalobacter) with diverse metabolic traits were retrieved from flooded rice roots. These findings suggest that endophytic communities could be significantly impacted by changes in plant-soil conditions derived from flooding during rice cropping. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  15. Time-course correlation of biofilm properties and electrochemical performance in single-chamber microbial fuel cells

    KAUST Repository

    Ren, Zhiyong; Ramasamy, Ramaraja P.; Cloud-Owen, Susan Red; Yan, Hengjing; Mench, Matthew M.; Regan, John M.

    2011-01-01

    The relationship between anode microbial characteristics and electrochemical parameters in microbial fuel cells (MFCs) was analyzed by time-course sampling of parallel single-bottle MFCs operated under identical conditions. While voltage stabilized within 4. days, anode biofilms continued growing during the six-week operation. Viable cell density increased asymptotically, but membrane-compromised cells accumulated steadily from only 9% of total cells on day 3 to 52% at 6. weeks. Electrochemical performance followed the viable cell trend, with a positive correlation for power density and an inverse correlation for anode charge transfer resistance. The biofilm architecture shifted from rod-shaped, dispersed cells to more filamentous structures, with the continuous detection of Geobacter sulfurreducens-like 16S rRNA fragments throughout operation and the emergence of a community member related to a known phenazine-producing Pseudomonas species. A drop in cathode open circuit potential between weeks two and three suggested that uncontrolled biofilm growth on the cathode deleteriously affects system performance. © 2010 Elsevier Ltd.

  16. Boosting biomethane yield and production rate with graphene: The potential of direct interspecies electron transfer in anaerobic digestion.

    Science.gov (United States)

    Lin, Richen; Cheng, Jun; Zhang, Jiabei; Zhou, Junhu; Cen, Kefa; Murphy, Jerry D

    2017-09-01

    Interspecies electron transfer between bacteria and archaea plays a vital role in enhancing energy efficiency of anaerobic digestion (AD). Conductive carbon materials (i.e. graphene nanomaterial and activated charcoal) were assessed to enhance AD of ethanol (a key intermediate product after acidogenesis of algae). The addition of graphene (1.0g/L) resulted in the highest biomethane yield (695.0±9.1mL/g) and production rate (95.7±7.6mL/g/d), corresponding to an enhancement of 25.0% in biomethane yield and 19.5% in production rate. The ethanol degradation constant was accordingly improved by 29.1% in the presence of graphene. Microbial analyses revealed that electrogenic bacteria of Geobacter and Pseudomonas along with archaea Methanobacterium and Methanospirillum might participate in direct interspecies electron transfer (DIET). Theoretical calculations provided evidence that graphene-based DIET can sustained a much higher electron transfer flux than conventional hydrogen transfer. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Microbial transformations of actinides in the environment

    Energy Technology Data Exchange (ETDEWEB)

    Livens, F R [Centre for Radiochemistry Research, School of Chemistry, University of Manchester, Oxford Road, Manchester M13 9PL (United Kingdom); Al-Bokari, M [Institute of Atomic Energy Research, King Abdulaziz City for Science and Technology, P. O. Box 6086, Riyadh 11442 (Saudi Arabia); Fomina, M; Gadd, G M [College of Life Sciences, University of Dundee, Dundee DD1 5EH (United Kingdom); Geissler, A; Lloyd, J R; Vaughan, D J [School of Earth, Atmospheric and Environmental Sciences, and Williamson Research Centre for Molecular Environmental Science, University of Manchester, Oxford Road, Manchester M13 9PL (United Kingdom); Renshaw, J C, E-mail: francis.livens@manchester.ac.uk [School of Geography, Earth and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT (United Kingdom)

    2010-03-15

    The diversity of microorganisms is still far from understood, although many examples of the microbial biotransformation of stable, pollutant and radioactive elements, involving Bacteria, Archaea and Fungi, are known. In estuarine sediments from the Irish Sea basin, which have been labelled by low level effluent discharges, there is evidence of an annual cycle in Pu solubility, and microcosm experiments have demonstrated both shifts in the bacterial community and changes in Pu solubility as a result of changes in redox conditions. In the laboratory, redox transformation of both U and Pu by Geobacter sulfurreducens has been demonstrated and EXAFS spectroscopy has been used to understand the inability of G. sufurreducens to reduce Np(V). Fungi promote corrosion of metallic U alloy through production of a range of carboxylic acid metabolites, and are capable of translocating the dissolved U before precipitating it externally to the hyphae, as U(VI) phosphate phases. These examples illustrate the far-reaching but complex effects which microorganisms can have on actinide behaviour.

  18. Retention and transport of an anaerobic trichloroethene dechlorinating microbial culture in anaerobic porous media.

    Science.gov (United States)

    Zhang, Huixin; Ulrich, Ania C; Liu, Yang

    2015-06-01

    The influence of solution chemistry on microbial transport was examined using the strictly anaerobic trichloroethene (TCE) bioaugmentation culture KB-1(®). A column was employed to determine transport behaviors and deposition kinetics of three distinct functional species in KB-1(®), Dehalococcoides, Geobacter, and Methanomethylovorans, over a range of ionic strengths under a well-controlled anaerobic condition. A quantitative polymerase chain reaction (qPCR) was utilized to enumerate cell concentration and complementary techniques were implemented to evaluate cell surface electrokinetic potentials. Solution chemistry was found to positively affect the deposition rates, which was consistent with calculated Derjaguin-Landau-Verwey-Overbeek (DLVO) interaction energies. Retained microbial profiles showed spatially constant colloid deposition rate coefficients, in agreement with classical colloid filtration theory (CFT). It was interesting to note that the three KB-1(®) species displayed similar transport and retention behaviors under the defined experimental conditions despite their different cell electrokinetic properties. A deeper analysis of cell characteristics showed that factors, such as cell size and shape, concentration, and motility were involved in determining adhesion behavior. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Multi-heme Cytochromes in Shewanella oneidensis MR-1: Structures, functions and opportunities

    Energy Technology Data Exchange (ETDEWEB)

    Breuer, Marian; Rosso, Kevin M.; Blumberger, Jochen; Butt, Julea N.

    2014-11-05

    Multi-heme cytochromes are employed by a range of microorganisms to transport electrons over distances of up to tens of nanometers. Perhaps the most spectacular utilization of these proteins is in the reduction of extracellular solid substrates, including electrodes and insoluble mineral oxides of Fe(III) and Mn(III/IV), by species of Shewanella and Geobacter. However, multi-heme cytochromes are found in numerous and phylogenetically diverse prokaryotes where they participate in electron transfer and redox catalysis that contributes to biogeochemical cycling of N, S and Fe on the global scale. These properties of multi-heme cytochromes have attracted much interest and contributed to advances in bioenergy applications and bioremediation of contaminated soils. Looking forward there are opportunities to engage multi-heme cytochromes for biological photovoltaic cells, microbial electrosynthesis and developing bespoke molecular devices. As a consequence it is timely to review our present understanding of these proteins and we do this here with a focus on the multitude of functionally diverse multi-heme cytochromes in Shewanella oneidensis MR-1. We draw on findings from experimental and computational approaches which ideally complement each other in the study of these systems: computational methods can interpret experimentally determined properties in terms of molecular structure to cast light on the relation between structure and function. We show how this synergy has contributed to our understanding of multi-heme cytochromes and can be expected to continue to do so for greater insight into natural processes and their informed exploitation in biotechnologies.

  20. High Performance Bioanode Development for Fermentable Substrates via Controlled Electroactive Biofilm Growth

    Energy Technology Data Exchange (ETDEWEB)

    Ichihashi, Osamu [ORNL; Vishnivetskaya, Tatiana A [ORNL; Borole, Abhijeet P [ORNL

    2014-11-11

    A bioanode was optimized to generate current densities reaching 38.4 4.9 A m-2, which brings bioelectrochemical systems closer to commercial consideration. Glucose and lactate were fed together in a continuous or fed-batch mode. The current density increased from 2.3 A m-2 to 38.4 A m-2 over a 33 day period and remained stable thereafter. The coulombic efficiency ranged from 50% to 80%. A change in substrate concentration from 200 mg L-1 to 5 mg L-1 decreased maximum current density from 38.4 A m-2 to 12.3 A m-2. The anode consortia included Firmicutes (55.0%), Proteobacteria (41.8%) and Bacteroidetes (2.1%) constituting two potential electrogenic genera: Geobacter (6.8%) and Aeromonas (31.9%). The current production was found to be limited by kinetics during the growth period (33 days), and mass transfer, thereafter. The results indicate the necessity of removing spent biomass for efficient long term operation and treatment of wastewater streams.

  1. Conductive properties of methanogenic biofilms.

    Science.gov (United States)

    Li, Cheng; Lesnik, Keaton Larson; Liu, Hong

    2018-02-01

    Extracellular electron transfer between syntrophic partners needs to be efficiently maintained in methanogenic environments. Direct extracellular electron transfer via electrical current is an alternative to indirect hydrogen transfer but requires construction of conductive extracellular structures. Conductive mechanisms and relationship between conductivity and the community composition in mixed-species methanogenic biofilms are not well understood. The present study investigated conductive behaviors of methanogenic biofilms and examined the correlation between biofilm conductivity and community composition between different anaerobic biofilms enriched from the same inoculum. Highest conductivity observed in methanogenic biofilms was 71.8±4.0μS/cm. Peak-manner response of conductivity upon changes over a range of electrochemical potentials suggests that electron transfer in methanogenic biofilms occurs through redox driven super-exchange. The strong correlation observed between biofilm conductivity and Geobacter spp. in the metabolically diverse anaerobic communities suggests that the efficiency of DEET may provide pressure for microbial communities to select for species that can produce electrical conduits. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    Jones, A.-Andrew; Buie, Cullen

    2016-11-01

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

  4. Novel RuCoSe as non-platinum catalysts for oxygen reduction reaction in microbial fuel cells

    Science.gov (United States)

    Rozenfeld, Shmuel; Schechter, Michal; Teller, Hanan; Cahan, Rivka; Schechter, Alex

    2017-09-01

    Microbial electrochemical cells (MECs) are explored for the conversion of acetate directly to electrical energy. This device utilizes a Geobacter sulfurreducens anode and a novel RuCoSe air cathode. RuCoSe synthesized in selected compositions by a borohydride reduction method produces amorphous structures of powdered agglomerates. Oxygen reduction reaction (ORR) was measured in a phosphate buffer solution pH 7 using a rotating disc electrode (RDE), from which the kinetic current (ik) was measured as a function of potential and composition. The results show that ik of RuxCoySe catalysts increases in the range of XRu = 0.25 > x > 0.7 and y < 0.15 for all tested potentials. A poisoning study of RuCoSe and Pt catalysts in a high concentration acetate solution shows improved tolerance of RuCoSe to this fuel at acetate concentration ≥500 mM. MEC discharge plots under physiological conditions show that ∼ RuCo2Se (sample S3) has a peak power density of 750 mW cm-2 which is comparable with Pt 900 mW cm-2.

  5. Enhanced bioelectricity generation of air-cathode buffer-free microbial fuel cells through short-term anolyte pH adjustment.

    Science.gov (United States)

    Ren, Yueping; Chen, Jinli; Li, Xiufen; Yang, Na; Wang, Xinhua

    2018-04-01

    Short-term initial anolyte pH adjustment can relieve the performance deterioration of the single-chamber air-cathode buffer-free microbial fuel cell (BFMFC) caused by anolyte acidification. Adjusting the initial anolyte pH to 9 in 5 running cycles is the optimum strategy. The relative abundance of the electrochemically active Geobacter in the KCl-pH9-MFC anode biofilm increased from 59.01% to 75.13% after the short-term adjustment. The maximum power density (P max ) of the KCl-pH9-MFC was elevated from 316.4mW·m -2 to 511.6mW·m -2 , which was comparable with that of the PBS-MFC. And, after the short-term adjusting, new equilibrium between the anolyte pH and the anode biofilm electrochemical activity has been established in the BFMFC, which ensured the sustainability of the improved bioelectricity generation performance. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Microbiome characterization of MFCs used for the treatment of swine manure.

    Science.gov (United States)

    Vilajeliu-Pons, Anna; Puig, Sebastià; Pous, Narcís; Salcedo-Dávila, Inmaculada; Bañeras, Lluís; Balaguer, Maria Dolors; Colprim, Jesús

    2015-05-15

    Conventional swine manure treatment is performed by anaerobic digestion, but nitrogen is not treated. Microbial Fuel Cells (MFCs) allow organic matter and nitrogen removal with concomitant electricity production. MFC microbiomes treating industrial wastewaters as swine manure have not been characterized. In this study, a multidisciplinary approach allowed microbiome relation with nutrient removal capacity and electricity production. Two different MFC configurations (C-1 and C-2) were used to treat swine manure. In C-1, the nitrification and denitrification processes took place in different compartments, while in C-2, simultaneous nitrification-denitrification occurred in the cathode. Clostridium disporicum and Geobacter sulfurreducens were identified in the anode compartments of both systems. C. disporicum was related to the degradation of complex organic matter compounds and G. sulfurreducens to electricity production. Different nitrifying bacteria populations were identified in both systems because of the different operational conditions. The highest microbial diversity was detected in cathode compartments of both configurations, including members of Bacteroidetes, Chloroflexiaceae and Proteobacteria. These communities allowed similar removal rates of organic matter (2.02-2.09 kg COD m(-3)d(-1)) and nitrogen (0.11-0.16 kg Nm(-3)d(-1)) in both systems. However, they differed in the generation of electric energy (20 and 2 mW m(-3) in C-1 and C-2, respectively). Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Biotechnology for uranium extraction and environmental control

    International Nuclear Information System (INIS)

    Natarajan, K.A.

    2012-01-01

    India is looking forward to augmenting mining and extraction of uranium mineral for its nuclear energy needs. Being a radio-active mineral, mining and processing of uranium ore deposits need be carried out in an environmentally acceptable fashion. In this respect, a biotechnological approach holds great promise since it is environment-friendly, cost-effective and energy-efficient. There are several types of microorganisms which inhabit uranium ore bodies and biogenesis plays an important role in the mineralisation and transport of uranium-bearing minerals under the earth's crust. Uranium occurrences in India are only meagre and it becomes essential to tap effectively all the available resources. Uraninite and pitchblende occurring along with sulfide mineralisation such as pyrite are ideal candidates for bioleaching. Acidithiobacillus ferrooxidans present ubiquitously in the ore deposits can be isolated, cultured and utilised to bring about efficient acidic dissolution of uranium. Many such commercial attempts to extract uranium from even lean ores using acidophilic autotrophic bacteria have been made in different parts of the world. Anaerobes such a Geobacter and Sulfate Reducing Bacteria (SRB) can be effectively used in uranium mining for environmental control. Radioactive uranium mined wastes and tailing dumps can be cleaned and protected using microorganisms. In this lecture use of biotechnology in uranium extraction and bioremediation is illustrated with practical examples. Applicability of environment-friendly biotechnology for mining and extraction of uranium from Indian deposits is outlined. Commercial potentials for bioremediation in uranium-containing wastes are emphasised. (author)

  8. Change in microbial communities in acetate- and glucose-fed microbial fuel cells in the presence of light

    KAUST Repository

    Xing, Defeng; Cheng, Shaoan; Regan, John M.; Logan, Bruce E.

    2009-01-01

    Power densities produced by microbial fuel cells (MFCs) in natural systems are changed by exposure to light through the enrichment of photosynthetic microorganisms. When MFCs with brush anodes were exposed to light (4000 lx), power densities increased by 8-10% for glucose-fed reactors, and 34% for acetate-fed reactors. Denaturing gradient gel electrophoresis (DGGE) profiles based on the 16S rRNA gene showed that exposure to high light levels changed the microbial communities on the anodes. Based on 16S rRNA gene clone libraries of light-exposed systems the anode communities using glucose were also significantly different than those fed acetate. Dominant bacteria that are known exoelectrogens were identified in the anode biofilm, including a purple nonsulfur (PNS) photosynthetic bacterium, Rhodopseudomonas palustris, and a dissimilatory iron-reducing bacterium, Geobacter sulfurreducens. Pure culture tests confirmed that PNS photosynthetic bacteria increased power production when exposed to high light intensities (4000 lx). These results demonstrate that power production and community composition are affected by light conditions as well as electron donors in single-chamber air-cathode MFCs. © 2009 Elsevier B.V. All rights reserved.

  9. Molecular and cultivation-dependent analysis of metal-reducing bacteria implicated in arsenic mobilisation in south-east asian aquifers

    International Nuclear Information System (INIS)

    Hery, Marina; Gault, Andrew G.; Rowland, Helen A.L.; Lear, Gavin; Polya, David A.; Lloyd, Jonathan R.

    2008-01-01

    The reduction of sorbed As(V) to the potentially more mobile As(III) by As-respiring anaerobic bacteria has been implicated in the mobilisation of the toxic metalloid in aquifer sediments in SE Asia. However, there is currently only a limited amount of information on the identity of the organisms that can respire As(V) in these sediment systems. Here experiments are described that have targeted As(V)-respiring bacteria using cultivation-independent molecular techniques, and also more traditional microbiological approaches that have used growth media highly selective for organisms that can grow using arsenate as the sole electron acceptor supplied for anaerobic growth. The molecular techniques used have initially targeted DNA from microcosms displaying maximal rates of arsenate reduction, both with and without added electron donor. More recent studies from the authors' laboratory have used stable isotope probing techniques, targeting DNA from the active microbial fraction in microcosms labelled with [ 13 C]acetate supplied as an electron donor for arsenate reduction. Phylogenetic analyses using a highly conserved genetic marker (the 16S rRNA gene) have suggested the involvement of Sulfurospirillum and Geobacter species in arsenate-respiration, and this has been supported further by complimentary experiments using more traditional microbiological techniques. Additional research required to clarify the role of these organisms in the mobilisation of As in situ are discussed

  10. Recycling of drinking water treatment residue as an additional medium in columns for effective P removal from eutrophic surface water.

    Science.gov (United States)

    Wang, Changhui; Wu, Yu; Bai, Leilei; Zhao, Yaqian; Yan, Zaisheng; Jiang, Helong; Liu, Xin

    2018-07-01

    This study assesses the feasibility of recycling drinking water treatment residue (DWTR) to treat eutrophic surface water in a one-year continuous flow column test. Heat-treated DWTR was used as an additional medium (2%-4%) in columns in case excessive organic matter and N were released from the DWTR to surface water. The results indicated that with minimal undesirable effects on other water properties, DWTR addition substantially enhanced P removal, rendering P concentrations in treated water oligotrophic and treated water unsuitable for Microcystis aeruginosa breeding. Long-term stable P removal by DWTR-column treatment was mainly attributed to the relatively low P levels in raw water (cycles and multiple pollution control (e.g., Dechloromonas, Geobacter, Leucobacter, Nitrospira, Rhodoplanes, and Sulfuritalea); an apparent decrease in Mycobacterium with potential pathogenicity was observed in DWTR-columns. Regardless, limited denitrification of DWTR-columns was observed as a result of low bioavailability of C in surface water. This finding indicates that DWTR can be used with other methods to ensure denitrification for enhanced treatment effects. Overall, the use of DWTR as an additional medium in column systems can potentially treat eutrophic surface water. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. Tolerance of anaerobic bacteria to chlorinated solvents.

    Science.gov (United States)

    Koenig, Joanna C; Groissmeier, Kathrin D; Manefield, Mike J

    2014-01-01

    The aim of this research was to evaluate the effects of four chlorinated aliphatic hydrocarbons (CAHs), perchloroethene (PCE), carbon tetrachloride (CT), chloroform (CF) and 1,2-dichloroethane (1,2-DCA), on the growth of eight anaerobic bacteria: four fermentative species (Escherichia coli, Klebsiella sp., Clostridium sp. and Paenibacillus sp.) and four respiring species (Pseudomonas aeruginosa, Geobacter sulfurreducens, Shewanella oneidensis and Desulfovibrio vulgaris). Effective concentrations of solvents which inhibited growth rates by 50% (EC50) were determined. The octanol-water partition coefficient or log Po/w of a CAH proved a generally satisfactory measure of its toxicity. Most species tolerated approximately 3-fold and 10-fold higher concentrations of the two relatively more polar CAHs CF and 1,2-DCA, respectively, than the two relatively less polar compounds PCE and CT. EC50 values correlated well with growth rates observed in solvent-free cultures, with fast-growing organisms displaying higher tolerance levels. Overall, fermentative bacteria were more tolerant to CAHs than respiring species, with iron- and sulfate-reducing bacteria in particular appearing highly sensitive to CAHs. These data extend the current understanding of the impact of CAHs on a range of anaerobic bacteria, which will benefit the field of bioremediation.

  12. Microbial community analysis of anaerobic reactors treating soft drink wastewater.

    Directory of Open Access Journals (Sweden)

    Takashi Narihiro

    Full Text Available The anaerobic packed-bed (AP and hybrid packed-bed (HP reactors containing methanogenic microbial consortia were applied to treat synthetic soft drink wastewater, which contains polyethylene glycol (PEG and fructose as the primary constituents. The AP and HP reactors achieved high COD removal efficiency (>95% after 80 and 33 days of the operation, respectively, and operated stably over 2 years. 16S rRNA gene pyrotag analyses on a total of 25 biofilm samples generated 98,057 reads, which were clustered into 2,882 operational taxonomic units (OTUs. Both AP and HP communities were predominated by Bacteroidetes, Chloroflexi, Firmicutes, and candidate phylum KSB3 that may degrade organic compound in wastewater treatment processes. Other OTUs related to uncharacterized Geobacter and Spirochaetes clades and candidate phylum GN04 were also detected at high abundance; however, their relationship to wastewater treatment has remained unclear. In particular, KSB3, GN04, Bacteroidetes, and Chloroflexi are consistently associated with the organic loading rate (OLR increase to 1.5 g COD/L-d. Interestingly, KSB3 and GN04 dramatically decrease in both reactors after further OLR increase to 2.0 g COD/L-d. These results indicate that OLR strongly influences microbial community composition. This suggests that specific uncultivated taxa may take central roles in COD removal from soft drink wastewater depending on OLR.

  13. Establishment and metabolic analysis of a model microbial community for understanding trophic and electron accepting interactions of subsurface anaerobic environments

    Directory of Open Access Journals (Sweden)

    Yang Zamin K

    2010-05-01

    Full Text Available Abstract Background Communities of microorganisms control the rates of key biogeochemical cycles, and are important for biotechnology, bioremediation, and industrial microbiological processes. For this reason, we constructed a model microbial community comprised of three species dependent on trophic interactions. The three species microbial community was comprised of Clostridium cellulolyticum, Desulfovibrio vulgaris Hildenborough, and Geobacter sulfurreducens and was grown under continuous culture conditions. Cellobiose served as the carbon and energy source for C. cellulolyticum, whereas D. vulgaris and G. sulfurreducens derived carbon and energy from the metabolic products of cellobiose fermentation and were provided with sulfate and fumarate respectively as electron acceptors. Results qPCR monitoring of the culture revealed C. cellulolyticum to be dominant as expected and confirmed the presence of D. vulgaris and G. sulfurreducens. Proposed metabolic modeling of carbon and electron flow of the three-species community indicated that the growth of C. cellulolyticum and D. vulgaris were electron donor limited whereas G. sulfurreducens was electron acceptor limited. Conclusions The results demonstrate that C. cellulolyticum, D. vulgaris, and G. sulfurreducens can be grown in coculture in a continuous culture system in which D. vulgaris and G. sulfurreducens are dependent upon the metabolic byproducts of C. cellulolyticum for nutrients. This represents a step towards developing a tractable model ecosystem comprised of members representing the functional groups of a trophic network.

  14. Microbial Reduction of Fe(III) and SO42- and Associated Microbial Communities in the Alluvial Aquifer Groundwater and Sediments.

    Science.gov (United States)

    Lee, Ji-Hoon; Lee, Bong-Joo

    2017-11-25

    Agricultural demands continuously increased use of groundwater, causing drawdown of water table and need of artificial recharge using adjacent stream waters. River water intrusion into groundwater can alter the geochemical and microbiological characteristics in the aquifer and subsurface. In an effort to investigate the subsurface biogeochemical activities before operation of artificial recharge at the test site, established at the bank of Nakdong River, Changwon, South Korea, organic carbon transported from river water to groundwater was mimicked and the effect on the indigenous microbial communities was investigated with the microcosm incubations of the groundwater and subsurface sediments. Laboratory incubations indicated microbial reduction of Fe(III) and sulfate. Next-generation Illumina MiSeq sequences of V4 region of 16S rRNA gene provided that the shifts of microbial taxa to Fe(III)-reducing and/or sulfate-reducing microorganisms such as Geobacter, Albidiferax, Desulfocapsa, Desulfuromonas, and Desulfovibrio were in good correlation with the sequential flourishment of microbial reduction of Fe(III) and sulfate as the incubations progressed. This suggests the potential role of dissolved organic carbons migrated with the river water into groundwater in the managed aquifer recharge system on the indigenous microbial community composition and following alterations of subsurface biogeochemistry and microbial metabolic activities.

  15. Electro-active bio-films: formation, characterization and mechanisms

    International Nuclear Information System (INIS)

    Parot, Sandrine

    2007-01-01

    Some bacteria, which are able to exchange electrons with a conductive material without mediator form on conductive surfaces electro-active bio-films. This bacterial property has been recently discovered (2001). Objectives of this work are to develop electro-active bio-films in various natural environments from indigenous flora, then through complementary electrochemical techniques (chrono-amperometry and cyclic voltammetry), to evaluate electro-activity of isolates coming from so-formed bio-films and to characterize mechanisms of electron transfer between bacteria and materials. First, electro-active bio-films have been developed under chrono-amperometry in garden compost and in water coming from Guyana mangrove. These bio-films were respectively able to use an electrode as electron acceptor (oxidation) or as electron donor (reduction). In compost, results obtained in chrono-amperometry and cyclic voltammetry suggest a two-step electron transfer: slow substrate consumption, then rapid electron transfer between bacteria and the electrode. Thereafter, the ability to reduce oxygen was demonstrated with cyclic voltammetry for facultative aerobic isolates from compost bio-films (Enterobacter spp. and Pseudomonas spp.) and for aerobic isolates obtained from marine electro-active bio-films (Roseobacter spp. in majority). Finally, bio-films inducing current increase in chrono-amperometry were developed in bioreactor with synthetic medium from a pure culture of isolates. Hence, for the first time, electro-activity of several anaerobic strains of Geobacter bremensis isolated from compost bio-films was highlighted. (author) [fr

  16. Application of meta-transcriptomics and –proteomics to analysis of in situ physiological state

    Directory of Open Access Journals (Sweden)

    Allan eKonopka

    2012-05-01

    Full Text Available Analysis of the growth-limiting factor or environmental stressors affecting microbes in situ is of fundamental importance but analytically difficult. Microbes can reduce in situ limiting nutrient concentrations to sub-micromolar levels, and contaminated ecosystems may contain multiple stressors. The patterns of gene or protein expression by microbes in nature can be used to infer growth limitations, because they are regulated in response to environmental conditions. Experimental studies under controlled conditions in the laboratory provide the physiological underpinnings for developing these physiological indicators. Although regulatory networks may differ among specific microbes, there are some broad principles that can be applied, related to limiting nutrient acquisition, resource allocation, and stress responses. As technologies for transcriptomics and proteomics mature, the capacity to apply these approaches to complex microbial communities will accelerate. In particular, global proteomics reflect expressed catalytic activities. Furthermore, the high mass accuracy of some proteomic approaches allows mapping back to specific microbial strains. For example, at the Rifle IFRC field site in Western Colorado, the physiological status of Fe(III-reducing populations has been tracked over time. Members of a subsurface clade within the Geobacter predominated during carbon amendment to the subsurface environment. At the functional level, proteomic identifications produced inferences regarding (i temporal changes in anabolism and catabolism of acetate, (ii the onset of N2 fixation when N became limiting, and (iii expression of phosphate transporters during periods of intense growth. The application of these approaches in situ can lead to discovery of novel physiological adaptations.

  17. Biological reduction of iron to the elemental state from ochre deposits of Skelton Beck in Northeast England

    Directory of Open Access Journals (Sweden)

    Pattanathu K S M Rahman

    2014-06-01

    Full Text Available Ochre, consequence of acid mine drainage, is iron oxides-rich soil pigments that can be found in the water drainage from historic base metal and coal mines. The anaerobic strains of Geobacter sulfurreducens and Shewanella denitrificans were used for the microbial reduction of iron from samples of ochre collected from Skelton Beck (Saltburn Orange River, NZ 66738 21588 in Northeast England. The aim of the research was to determine the ability of the two anaerobic bacteria to reduce the iron present in ochre and to determine the rate of the reduction process. The physico-chemical changes in the ochre sample after the microbial reduction process were observed by the production of zero-valent iron which was later confirmed by the detection of elemental Fe in XRD spectrum. The XRF results revealed that 69.16% and 84.82% of iron oxide can be reduced using G. sulfurreducens and S. denitrificans respectively after 8 days of incubation. These results could provide the basis for the development of a biohydrometallurgical process for the production of elemental iron from ochre sediments.

  18. Response of methane production via propionate oxidation to carboxylated multiwalled carbon nanotubes in paddy soil enrichments

    Directory of Open Access Journals (Sweden)

    Jianchao Zhang

    2018-01-01

    Full Text Available Carboxylated multiwalled carbon nanotubes (MWCNTs-COOH have become a growing concern in terms of their fate and toxicity in aqueous environments. Methane (CH4 is a major product of organic matter degradation in waterlogged environments. In this study, we determined the effect of MWCNTs-COOH on the production of CH4 from propionate oxidation in paddy soil enrichments. The results showed that the methanogenesis from propionate degradation was accelerated in the presence of MWCNTs-COOH. In addition, the rates of CH4 production and propionate degradation increased with increasing concentrations of MWCNTs-COOH. Scanning electron microscopy (SEM observations showed that the cells were intact and maintained their structure in the presence of MWCNTs-COOH. In addition, SEM and fluorescence in situ hybridization (FISH images revealed that the cells were in direct contact with the MWCNTs and formed cell-MWCNTs aggregates that contained both bacteria and archaea. On the other hand, nontoxic magnetite nanoparticles (Fe3O4 had similar effects on the CH4 production and cell integrity as the MWCNTs-COOH. Compared with no nanomaterial addition, the relative abundances of Geobacter and Methanosarcina species increased in the presence of MWCNTs-COOH. This study suggests that MWCNTs-COOH exerted positive rather than cytotoxic effects on the syntrophic oxidation of propionate in paddy soil enrichments and affected the bacterial and archaeal community structure at the test concentrations. These findings provide novel insight into the consequences of nanomaterial release into anoxic natural environments.

  19. Diversity and function of the microbial community on anodes of sediment microbial fuel cells fueled by root exudates

    Energy Technology Data Exchange (ETDEWEB)

    Cabezas da Rosa, Angela

    2010-11-26

    Anode microbial communities are essential for current production in microbial fuel cells. Anode reducing bacteria are capable of using the anode as final electron acceptor in their respiratory chain. The electrons delivered to the anode travel through a circuit to the cathode where they reduce oxygen to water generating an electric current. A novel type of sediment microbial fuel cell (SMFC) harvest energy from photosynthetically derived compounds released through the roots. Nothing is known about anode microbial communities of this type of microbial fuel cell. This work consists of three parts. The first part focuses on the study of bacterial and archaeal community compositions on anodes of SMFCs fueled by rice root exudates. By using terminal restriction fragment length polymorphism (T-RFLP), a profiling technique, and cloning / sequencing of 16S rRNA, we determined that the support type used for the plant (vermiculite, potting soil or rice field soil) is an important factor determining the composition of the microbial community. Finally, by comparing microbial communities of current producing anodes and non-current producing controls we determined that Desulfobulbus- and Geobacter-related populations were probably most important for current production in potting soil and rice field soil SMFCs, respectively. However, {delta}-proteobacterial Anaeromyxobacter spp., unclassified {delta}-proteobacteria and Anaerolineae were also part of the anode biofilm in rice field soil SMFCs and these populations might also play a role in current production. Moreover, distinct clusters of Geobacter and Anaeromyxobacter populations were stimulated by rice root exudates. Regarding Archaea, uncultured Euryarchaea were abundant on anodes of potting soil SMFCs indicating a potential role in current production. In both, rice field soil and potting soil SMFCs, a decrease of Methanosaeta, an acetotrophic methanogen, was detected on current producing anodes. In the second part we focused

  20. Uranium Immobilization in Wetland Soils

    Science.gov (United States)

    Jaffe, Peter R.; Koster van Groos, Paul G.; Li, Dien; Chang, Hyun-Shik; Seaman, John C.; Kaplan, Daniel I.; Peacock, Aaron D.; Scheckel, Kirk

    2014-05-01

    In wetlands, which are a major feature at the groundwater-surface water interface, plants deliver oxygen to the subsurface to keep root tissue aerobic. Some of this oxygen leaches into the rhizosphere where it will oxidize iron that typically precipitates on or near roots. Furthermore, plans provide carbon via root exudates and turnover, which in the presence of the iron oxides drives the activity of heterotrophic iron reducers in wetland soils. Oxidized iron is an important electron acceptor for many microbially-driven transformations, which can affect the fate and transport of several pollutants. It has been shown that heterotrophic iron reducing organisms, such as Geobacter sp., can reduce water soluble U(VI) to insoluble U(IV). The goal of this study was to determine if and how iron cycling in the wetland rhizosphere affects uranium dynamics. For this purpose, we operated a series of small-scale wetland mesocosms in a greenhouse to simulate the discharge of uranium-contaminated groundwater to surface waters. The mesocosms were operated with two different Fe(II) loading rates, two plant types, and unplanted controls. The mesocosms contained zones of root exclusion to differentiate between the direct presence and absence of roots in the planted mesocosms. The mesocosms were operated for several month to get fully established, after which a U(VI) solution was fed for 80 days. The mesocosms were then sacrificed and analyzed for solid-associated chemical species, microbiological characterization, micro-X-ray florescence (µ-XRF) mapping of Fe and U on the root surface, and U speciation via X-ray Absorption Near Edge Structure (XANES). Results showed that bacterial numbers including Geobacter sp., Fe(III), as well as total uranium, were highest on roots, followed by sediments near roots, and lowest in zones without much root influence. Results from the µ-XRF mapping on root surfaces indicated a strong spatial correlation between Fe and U. This correlation was

  1. Strong Regionality and Dominance of Anaerobic Bacterial Taxa Characterize Diazotrophic Bacterial Communities of the Arcto-Alpine Plant Species Oxyria digyna and Saxifraga oppositifolia.

    Science.gov (United States)

    Kumar, Manoj; van Elsas, Jan Dirk; Nissinen, Riitta

    2017-01-01

    Arctic and alpine biomes are most often strongly nitrogen-limited, and hence biological nitrogen fixation is a strong driver of these ecosystems. Both biomes are characterized by low temperatures and short growing seasons, but they differ in seasonality of solar radiation and in soil water balance due to underlying permafrost in the Arctic. Arcto-alpine plant species are well-adapted to the low temperatures that prevail in their habitats, and plant growth is mainly limited by the availability of nutrients, in particular nitrogen, due to slow mineralization. Nitrogen fixing bacteria are likely important for plant growth in these habitats, but very little is known of these bacteria or forces shaping their communities. In this study, we characterized the potential nitrogen fixing bacterial (PNFB) communities associated with two arcto-alpine pioneer plant species, Oxyria digyna (mountain sorrel) and Saxifraga oppositifolia (blue saxifrage), in three climate regions. Both of these plants readily colonize low nutrient mineral soils. Our goal was to investigate how climate (region) and, on the other hand, host plant and plant species shape these communities. To our knowledge, this is the first comprehensive study describing PNFB communities associated with pioneer plants in different arcto-alpine biomes. Replicate samples were taken from two arctic regions, Kilpisjärvi and Ny-Ålesund, and one alpine region, Mayrhofen. In these, the PNFB communities in the bulk and rhizosphere soils and the plant endospheres were characterized by nifH -targeted PCR and massive parallel sequencing. The data revealed strong effects of climatic region on the dominating nitrogen fixers. Specifically, nifH sequences related to Geobacter (δ- Proteobacteria ) were present in high relative abundances in the nitrogen-fixing communities in the Mayrhofen and Kilpisjärvi regions, while members of the Clostridiales prevailed in the Kilpisjärvi and Ny-Ålesund regions. The bulk and rhizosphere soil

  2. Strong Regionality and Dominance of Anaerobic Bacterial Taxa Characterize Diazotrophic Bacterial Communities of the Arcto-Alpine Plant Species Oxyria digyna and Saxifraga oppositifolia

    Directory of Open Access Journals (Sweden)

    Manoj Kumar

    2017-10-01

    Full Text Available Arctic and alpine biomes are most often strongly nitrogen-limited, and hence biological nitrogen fixation is a strong driver of these ecosystems. Both biomes are characterized by low temperatures and short growing seasons, but they differ in seasonality of solar radiation and in soil water balance due to underlying permafrost in the Arctic. Arcto-alpine plant species are well-adapted to the low temperatures that prevail in their habitats, and plant growth is mainly limited by the availability of nutrients, in particular nitrogen, due to slow mineralization. Nitrogen fixing bacteria are likely important for plant growth in these habitats, but very little is known of these bacteria or forces shaping their communities. In this study, we characterized the potential nitrogen fixing bacterial (PNFB communities associated with two arcto-alpine pioneer plant species, Oxyria digyna (mountain sorrel and Saxifraga oppositifolia (blue saxifrage, in three climate regions. Both of these plants readily colonize low nutrient mineral soils. Our goal was to investigate how climate (region and, on the other hand, host plant and plant species shape these communities. To our knowledge, this is the first comprehensive study describing PNFB communities associated with pioneer plants in different arcto-alpine biomes. Replicate samples were taken from two arctic regions, Kilpisjärvi and Ny-Ålesund, and one alpine region, Mayrhofen. In these, the PNFB communities in the bulk and rhizosphere soils and the plant endospheres were characterized by nifH-targeted PCR and massive parallel sequencing. The data revealed strong effects of climatic region on the dominating nitrogen fixers. Specifically, nifH sequences related to Geobacter (δ-Proteobacteria were present in high relative abundances in the nitrogen-fixing communities in the Mayrhofen and Kilpisjärvi regions, while members of the Clostridiales prevailed in the Kilpisjärvi and Ny-Ålesund regions. The bulk and

  3. Dissecting the functional role of key residues in triheme cytochrome PpcA: a path to rational design of G. sulfurreducens strains with enhanced electron transfer capabilities.

    Directory of Open Access Journals (Sweden)

    Leonor Morgado

    Full Text Available PpcA is the most abundant member of a family of five triheme cytochromes c7 in the bacterium Geobacter sulfurreducens (Gs and is the most likely carrier of electrons destined for outer surface during respiration on solid metal oxides, a process that requires extracellular electron transfer. This cytochrome has the highest content of lysine residues (24% among the family, and it was suggested to be involved in e-/H(+ energy transduction processes. In the present work, we investigated the functional role of lysine residues strategically located in the vicinity of each heme group. Each lysine was replaced by glutamine or glutamic acid to evaluate the effects of a neutral or negatively charged residue in each position. The results showed that replacing Lys9 (located near heme IV, Lys18 (near heme I or Lys22 (between hemes I and III has essentially no effect on the redox properties of the heme groups and are probably involved in redox partner recognition. On the other hand, Lys43 (near heme IV, Lys52 (between hemes III and IV and Lys60 (near heme III are crucial in the regulation of the functional mechanism of PpcA, namely in the selection of microstates that allow the protein to establish preferential e-/H(+ transfer pathways. The results showed that the preferred e-/H(+ transfer pathways are only established when heme III is the last heme to oxidize, a feature reinforced by a higher difference between its reduction potential and that of its predecessor in the order of oxidation. We also showed that K43 and K52 mutants keep the mechanistic features of PpcA by establishing preferential e-/H+ transfer pathways at lower reduction potential values than the wild-type protein, a property that can enable rational design of Gs strains with optimized extracellular electron transfer capabilities.

  4. Responses of microbial community functional structures to pilot-scale uranium in situ bioremediation

    Energy Technology Data Exchange (ETDEWEB)

    Xu, M.; Wu, W.-M.; Wu, L.; He, Z.; Van Nostrand, J.D.; Deng, Y.; Luo, J.; Carley, J.; Ginder-Vogel, M.; Gentry, T.J.; Gu, B.; Watson, D.; Jardine, P.M.; Marsh, T.L.; Tiedje, J.M.; Hazen, T.C.; Criddle, C.S.; Zhou, J.

    2010-02-15

    A pilot-scale field test system with an inner loop nested within an outer loop was constructed for in situ U(VI) bioremediation at a US Department of Energy site, Oak Ridge, TN. The outer loop was used for hydrological protection of the inner loop where ethanol was injected for biostimulation of microorganisms for U(VI) reduction/immobilization. After 2 years of biostimulation with ethanol, U(VI) levels were reduced to below drinking water standard (<30 {micro}gl{sup -1}) in the inner loop monitoring wells. To elucidate the microbial community structure and functions under in situ uranium bioremediation conditions, we used a comprehensive functional gene array (GeoChip) to examine the microbial functional gene composition of the sediment samples collected from both inner and outer loop wells. Our study results showed that distinct microbial communities were established in the inner loop wells. Also, higher microbial functional gene number, diversity and abundance were observed in the inner loop wells than the outer loop wells. In addition, metal-reducing bacteria, such as Desulfovibrio, Geobacter, Anaeromyxobacter and Shewanella, and other bacteria, for example, Rhodopseudomonas and Pseudomonas, are highly abundant in the inner loop wells. Finally, the richness and abundance of microbial functional genes were highly correlated with the mean travel time of groundwater from the inner loop injection well, pH and sulfate concentration in groundwater. These results suggest that the indigenous microbial communities can be successfully stimulated for U bioremediation in the groundwater ecosystem, and their structure and performance can be manipulated or optimized by adjusting geochemical and hydrological conditions.

  5. Transport of poly(acrylic acid) coated 2-line ferrihydrite nanoparticles in saturated aquifer sediments for environmental remediation

    Science.gov (United States)

    Xiang, Aishuang; Zhou, Sheng; Koel, Bruce E.; Jaffé, Peter R.

    2014-04-01

    Groundwater remediation using iron oxide and zero-valent iron nanoparticles (NPs) can be effective, but is limited in many applications due to the NP strong retention in groundwater-saturated porous media after injection, the passivation of the porous surface, and the high cost of nanomaterials versus macro scale iron. In this study, we investigated transport of bare and polymer-coated 2-line ferrihydrite NPs (30-300 nm) in saturated aquifer sediments. The influence of poly(acrylic acid) (PAA) polymer coatings was studied on the colloidal stability and transport in sediments packed column tests simulating groundwater flow in saturated sediments. In addition, the influence of calcium cations was investigated by transport measurements using sediments with calcium concentrations in the aqueous phase ranging from 0.5 (typical for most sediments) to 2 mM. Measurements were also made of zeta potential, hydrodynamic diameter, polymer adsorption and desorption properties, and bio-availability of PAA-coated NPs. We found that NP transport through the saturated aquifer sediments was improved by PAA coating and that the transport properties could be tuned by adjusting the polymer concentration. We further discovered that PAA coatings enhanced NP transport, compared to bare NPs, in all calcium-containing experiments tested, however, the presence of calcium always exhibited a negative effect on NP transport. In tests of bioavailability, the iron reduction rate of the coated and bare NPs by Geobacter sulfurreducens was the same, which shows that the PAA coating does not significantly reduce NP Fe(III) bioavailability. Our results demonstrate that much improved transport of iron oxide NP can be achieved in saturated aquifer sediments by introducing negatively charged polyelectrolytes and optimizing polymer concentrations, and furthermore, these coated NPs retain their bioavailability that is needed for applications in bio-environmental remediation.

  6. Phase Preference by Active, Acetate-Utilizing Bacteria at the Rifle, CO Integrated Field Research Challenge Site

    International Nuclear Information System (INIS)

    Kerkhoff, Lee; Williams, Kenneth H.; Long, Philip E.; McGuinness, L.

    2011-01-01

    Uranium contaminated groundwaters are a legacy concern for the U.S. Department of Energy. Previous experiments at the Rifle, Colorado Integrated Field Challenge (IFC) site have demonstrated that field-scale addition of acetate to groundwater reduces the ambient soluable uranium concentration, sequestering the radionuclide as uraninite. However, questions remain regarding which microorganism(s) are consuming this acetate and if active groundwater microorganisms are different from active particle-associated bacteria. In this report, 13-C acetate was used to assess the active microbes that synthesize DNA on 3 size fractions (coarse sand, fines (8-approximately 150 micron), groundwater (0.2-8 micron)) over a 24 -day time frame. Results indicated a stronger signal from 13-C acetate associated with the 'fines' fraction compared with smaller amounts of 13-C uptake on the sand fraction and groundwater samples during the SIP incubations. TRFLP analysis of this 13-C-labeled DNA, indicated 31+ 9 OTU's with 6 peaks dominating the active profiles (166, 187, 210, 212, and 277 bp peaks using MnlI). Cloning/sequencing of the amplification products indicated a Geobacter-like group (187, 210, 212 bp) primarily synthesized DNA from acetate in the groundwater phase, an alpha Proteobacterium (166 bp) primarily grew on the fines/sands, and an Acinetobacter sp. (277 bp) utilized much of the 13C acetate in both groundwater and particle-associated phases. These findings will help to delineate the acetate utilization patterns of bacteria during field-scale acetate addition and can lead to improved methods for stimulating distinct microbial populations in situ.

  7. Cathodic microbial community adaptation to the removal of chlorinated herbicide in soil microbial fuel cells.

    Science.gov (United States)

    Li, Yue; Li, Xiaojing; Sun, Yang; Zhao, Xiaodong; Li, Yongtao

    2018-04-05

    The microbial fuel cell (MFC) that uses a solid electrode as the inexhaustible electron acceptor is an innovative remediation technology that simultaneously generates bioelectricity. Chlorinated pollutants are better metabolized by reductive dechlorination in proximity to the cathode. Here, the removal efficiency of the herbicide metolachlor (ML) increased by 262 and 176% in soil MFCs that were spiked with 10 (C10) and 20 mg/kg (C20) of ML, respectively, relative to the non-electrode controls. The bioelectricity output of the C10 and C20 increased by over two- and eightfold, respectively, compared to that of the non-ML control, with maximum current densities of 49.6 ± 2.5 (C10) and 78.9 ± 0.6 mA/m 2 (C20). Based on correlations between ML concentrations and species abundances in the MFCs, it was inferred that Azohydromonas sp., Sphingomonas sp., and Pontibacter sp. play a major role in ML removal around the cathode, with peak removal efficiencies of 56 ± 1% (C10) and 58 ± 1% (C20). Moreover, Clostridium sp., Geobacter sp., Bacillus sp., Romboutsia sp., and Terrisporobacter sp. may be electricigens or closely related microbes due to the significant positive correlation between the bioelectricity generation levels and their abundances around the anode. This study suggests that a directional adaptation of the microbial community has taken place to increase both the removal of chlorinated herbicides around the cathode and the generation of bioelectricity around the anode in bioelectrochemical remediation systems.

  8. Significant Association between Sulfate-Reducing Bacteria and Uranium-Reducing Microbial Communities as Revealed by a Combined Massively Parallel Sequencing-Indicator Species Approach▿ †

    Science.gov (United States)

    Cardenas, Erick; Wu, Wei-Min; Leigh, Mary Beth; Carley, Jack; Carroll, Sue; Gentry, Terry; Luo, Jian; Watson, David; Gu, Baohua; Ginder-Vogel, Matthew; Kitanidis, Peter K.; Jardine, Philip M.; Zhou, Jizhong; Criddle, Craig S.; Marsh, Terence L.; Tiedje, James M.

    2010-01-01

    Massively parallel sequencing has provided a more affordable and high-throughput method to study microbial communities, although it has mostly been used in an exploratory fashion. We combined pyrosequencing with a strict indicator species statistical analysis to test if bacteria specifically responded to ethanol injection that successfully promoted dissimilatory uranium(VI) reduction in the subsurface of a uranium contamination plume at the Oak Ridge Field Research Center in Tennessee. Remediation was achieved with a hydraulic flow control consisting of an inner loop, where ethanol was injected, and an outer loop for flow-field protection. This strategy reduced uranium concentrations in groundwater to levels below 0.126 μM and created geochemical gradients in electron donors from the inner-loop injection well toward the outer loop and downgradient flow path. Our analysis with 15 sediment samples from the entire test area found significant indicator species that showed a high degree of adaptation to the three different hydrochemical-created conditions. Castellaniella and Rhodanobacter characterized areas with low pH, heavy metals, and low bioactivity, while sulfate-, Fe(III)-, and U(VI)-reducing bacteria (Desulfovibrio, Anaeromyxobacter, and Desulfosporosinus) were indicators of areas where U(VI) reduction occurred. The abundance of these bacteria, as well as the Fe(III) and U(VI) reducer Geobacter, correlated with the hydraulic connectivity to the substrate injection site, suggesting that the selected populations were a direct response to electron donor addition by the groundwater flow path. A false-discovery-rate approach was implemented to discard false-positive results by chance, given the large amount of data compared. PMID:20729318

  9. Significant association between sulfate-reducing bacteria and uranium-reducing microbial communities as revealed by a combined massively parallel sequencing-indicator species approach.

    Science.gov (United States)

    Cardenas, Erick; Wu, Wei-Min; Leigh, Mary Beth; Carley, Jack; Carroll, Sue; Gentry, Terry; Luo, Jian; Watson, David; Gu, Baohua; Ginder-Vogel, Matthew; Kitanidis, Peter K; Jardine, Philip M; Zhou, Jizhong; Criddle, Craig S; Marsh, Terence L; Tiedje, James M

    2010-10-01

    Massively parallel sequencing has provided a more affordable and high-throughput method to study microbial communities, although it has mostly been used in an exploratory fashion. We combined pyrosequencing with a strict indicator species statistical analysis to test if bacteria specifically responded to ethanol injection that successfully promoted dissimilatory uranium(VI) reduction in the subsurface of a uranium contamination plume at the Oak Ridge Field Research Center in Tennessee. Remediation was achieved with a hydraulic flow control consisting of an inner loop, where ethanol was injected, and an outer loop for flow-field protection. This strategy reduced uranium concentrations in groundwater to levels below 0.126 μM and created geochemical gradients in electron donors from the inner-loop injection well toward the outer loop and downgradient flow path. Our analysis with 15 sediment samples from the entire test area found significant indicator species that showed a high degree of adaptation to the three different hydrochemical-created conditions. Castellaniella and Rhodanobacter characterized areas with low pH, heavy metals, and low bioactivity, while sulfate-, Fe(III)-, and U(VI)-reducing bacteria (Desulfovibrio, Anaeromyxobacter, and Desulfosporosinus) were indicators of areas where U(VI) reduction occurred. The abundance of these bacteria, as well as the Fe(III) and U(VI) reducer Geobacter, correlated with the hydraulic connectivity to the substrate injection site, suggesting that the selected populations were a direct response to electron donor addition by the groundwater flow path. A false-discovery-rate approach was implemented to discard false-positive results by chance, given the large amount of data compared.

  10. Chlorine isotope effects from isotope ratio mass spectrometry suggest intramolecular C-Cl bond competition in trichloroethene (TCE) reductive dehalogenation.

    Science.gov (United States)

    Cretnik, Stefan; Bernstein, Anat; Shouakar-Stash, Orfan; Löffler, Frank; Elsner, Martin

    2014-05-20

    Chlorinated ethenes are prevalent groundwater contaminants. To better constrain (bio)chemical reaction mechanisms of reductive dechlorination, the position-specificity of reductive trichloroethene (TCE) dehalogenation was investigated. Selective biotransformation reactions (i) of tetrachloroethene (PCE) to TCE in cultures of Desulfitobacterium sp. strain Viet1; and (ii) of TCE to cis-1,2-dichloroethene (cis-DCE) in cultures of Geobacter lovleyi strain SZ were investigated. Compound-average carbon isotope effects were -19.0‰ ± 0.9‰ (PCE) and -12.2‰ ± 1.0‰ (TCE) (95% confidence intervals). Using instrumental advances in chlorine isotope analysis by continuous flow isotope ratio mass spectrometry, compound-average chorine isotope effects were measured for PCE (-5.0‰ ± 0.1‰) and TCE (-3.6‰ ± 0.2‰). In addition, position-specific kinetic chlorine isotope effects were determined from fits of reactant and product isotope ratios. In PCE biodegradation, primary chlorine isotope effects were substantially larger (by -16.3‰ ± 1.4‰ (standard error)) than secondary. In TCE biodegradation, in contrast, the product cis-DCE reflected an average isotope effect of -2.4‰ ± 0.3‰ and the product chloride an isotope effect of -6.5‰ ± 2.5‰, in the original positions of TCE from which the products were formed (95% confidence intervals). A greater difference would be expected for a position-specific reaction (chloride would exclusively reflect a primary isotope effect). These results therefore suggest that both vicinal chlorine substituents of TCE were reactive (intramolecular competition). This finding puts new constraints on mechanistic scenarios and favours either nucleophilic addition by Co(I) or single electron transfer as reductive dehalogenation mechanisms.

  11. Effects of 1,1,1-Trichloroethane and Triclocarban on Reductive Dechlorination of Trichloroethene in a TCE-Reducing Culture.

    Science.gov (United States)

    Wen, Li-Lian; Chen, Jia-Xian; Fang, Jia-Yi; Li, Ang; Zhao, He-Ping

    2017-01-01

    Chlorinated compounds were generally present in the environment due to widespread use in the industry. A short-term study was performed to evaluate the effects of 1,1,1- trichloroethane (TCA) and triclocarban (TCC) on trichloroethene (TCE) removal in a reactor fed with lactate as the sole electron donor. Both TCA and TCC inhibited TCE reduction, but the TCC had a more pronounced effect compared to TCA. The TCE-reducing culture, which had never been exposed to TCA before, reductively dechlorinated TCA to 1,1-dichloroethane (DCA). Below 15 μM, TCA had little effect on the transformation of TCE to cis -dichloroethene (DCE); however, the reduction of cis -DCE and vinyl chloride (VC) were more sensitive to TCA, and ethene production was completely inhibited when the concentration of TCA was above 15 μM. In cultures amended with TCC, the reduction of TCE was severely affected, even at concentrations as low as 0.3 μM; all the cultures stalled at VC, and no ethene was detected. The cultures that fully transformed TCE to ethene contained 5.2-8.1% Dehalococcoides . Geobacter and Desulfovibrio , the bacteria capable of partially reducing TCE to DCE, were detected in all cultures, but both represented a larger proportion of the community in TCC-amended cultures. All cultures were dominated by Clostridium _sensu_stricto_7, a genus that belongs to Firmicutes with proportions ranging from 40.9% (in a high TCC (15 μM) culture) to 88.2%. Methanobacteria was detected at levels of 1.1-12.7%, except in cultures added with 15 and 30 μM TCA, in which they only accounted for ∼0.4%. This study implies further environmental factors needed to be considered in the successful bioremediation of TCE in contaminated sites.

  12. Optimizing Cr(VI) and Tc(VII) remediation through nano-scale biomineral engineering

    International Nuclear Information System (INIS)

    Cutting, R.S.; Coker, V.S.; Telling, N.D.; Kimber, R.L.; Pearce, C.I.; Ellis, B.; Lawson, R; van der Laan, G.; Pattrick, R.A.D.; Vaughan, D.J.; Arenholz, E.; Lloyd, J.R.

    2009-01-01

    To optimize the production of biomagnetite for the bioremediation of metal oxyanion contaminated waters, the reduction of aqueous Cr(VI) to Cr(III) by two biogenic magnetites and a synthetic magnetite was evaluated under batch and continuous flow conditions. Results indicate that nano-scale biogenic magnetite produced by incubating synthetic schwertmannite powder in cell suspensions of Geobacter sulfurreducens is more efficient at reducing Cr(VI) than either biogenic nano-magnetite produced from a suspension of ferrihydrite 'gel' or synthetic nano-scale Fe 3 O 4 powder. Although X-ray Photoelectron Spectroscopy (XPS) measurements obtained from post-exposure magnetite samples reveal that both Cr(III) and Cr(VI) are associated with nanoparticle surfaces, X-ray Magnetic Circular Dichroism (XMCD) studies indicate that some Cr(III) has replaced octahedrally coordinated Fe in the lattice of the magnetite. Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES) measurements of total aqueous Cr in the associated solution phase indicated that, although the majority of Cr(III) was incorporated within or adsorbed to the magnetite samples, a proportion (∼10-15 %) was released back into solution. Studies of Tc(VII) uptake by magnetites produced via the different synthesis routes also revealed significant differences between them as regards effectiveness for remediation. In addition, column studies using a γ-camera to obtain real time images of a 99m Tc(VII) radiotracer were performed to visualize directly the relative performances of the magnetite sorbents against ultra-trace concentrations of metal oxyanion contaminants. Again, the magnetite produced from schwertmannite proved capable of retaining more (∼20%) 99m Tc(VII) than the magnetite produced from ferrihydrite, confirming that biomagnetite production for efficient environmental remediation can be fine-tuned through careful selection of the initial Fe(III) mineral substrate supplied to Fe

  13. Abiotic versus biotic iron mineral transformation studied by a miniaturized backscattering Mössbauer spectrometer (MIMOS II), X-ray diffraction and Raman spectroscopy

    Science.gov (United States)

    Markovski, C.; Byrne, J. M.; Lalla, E.; Lozano-Gorrín, A. D.; Klingelhöfer, G.; Rull, F.; Kappler, A.; Hoffmann, T.; Schröder, C.

    2017-11-01

    Searching for biomarkers or signatures of microbial transformations of minerals is a critical aspect for determining how life evolved on Earth, and whether or not life may have existed in other planets, including Mars. In order to solve such questions, several missions to Mars have sought to determine the geochemistry and mineralogy on the Martian surface. This research includes the two miniaturized Mössbauer spectrometers (MIMOS II) on board the Mars Exploration Rovers Spirit and Opportunity, which have detected a variety of iron minerals on Mars, including magnetite (Fe2+Fe3+2O4) and goethite (α-FeO(OH)). On Earth, both minerals can derive from microbiological activity (e.g. through dissimilatory iron reduction of ferrihydrite by Fe(III)-reducing bacteria). Here we used a lab based MIMOS II to characterize the mineral products of biogenic transformations of ferrihydrite to magnetite by the Fe(III)-reducing bacteria Geobacter sulfurreducens. In combination with Raman spectroscopy and X-ray diffraction (XRD), we observed the formation of magnetite, goethite and siderite. We compared the material produced by biogenic transformations to abiotic samples in order to distinguish abiotic and biotic iron minerals by techniques that are or will be available onboard Martian based laboratories. The results showed the possibility to distinguish the abiotic and biotic origin of the minerals. Mossbauer was able to distinguish the biotic/abiotic magnetite with the interpretation of the geological context (Fe content mineral assemblages and accompanying minerals) and the estimation of the particle size in a non-destructive way. The Raman was able to confirm the biotic/abiotic principal peaks of the magnetite, as well as the organic principal vibration bands attributed to the bacteria. Finally, the XRD confirmed the particle size and mineralogy.

  14. Arsenic(V) reduction in relation to Iron(III) transformation and molecular characterization of the structural and functional microbial community in sediments of a basin-fill aquifer in Northern Utah.

    Science.gov (United States)

    Mirza, Babur S; Muruganandam, Subathra; Meng, Xianyu; Sorensen, Darwin L; Dupont, R Ryan; McLean, Joan E

    2014-05-01

    Basin-fill aquifers of the Southwestern United States are associated with elevated concentrations of arsenic (As) in groundwater. Many private domestic wells in the Cache Valley Basin, UT, have As concentrations in excess of the U.S. EPA drinking water limit. Thirteen sediment cores were collected from the center of the valley at the depth of the shallow groundwater and were sectioned into layers based on redoxmorphic features. Three of the layers, two from redox transition zones and one from a depletion zone, were used to establish microcosms. Microcosms were treated with groundwater (GW) or groundwater plus glucose (GW+G) to investigate the extent of As reduction in relation to iron (Fe) transformation and characterize the microbial community structure and function by sequencing 16S rRNA and arsenate dissimilatory reductase (arrA) genes. Under the carbon-limited conditions of the GW treatment, As reduction was independent of Fe reduction, despite the abundance of sequences related to Geobacter and Shewanella, genera that include a variety of dissimilatory iron-reducing bacteria. The addition of glucose, an electron donor and carbon source, caused substantial shifts toward domination of the bacterial community by Clostridium-related organisms, and As reduction was correlated with Fe reduction for the sediments from the redox transition zone. The arrA gene sequencing from microcosms at day 54 of incubation showed the presence of 14 unique phylotypes, none of which were related to any previously described arrA gene sequence, suggesting a unique community of dissimilatory arsenate-respiring bacteria in the Cache Valley Basin.

  15. Biotransformations Involved in Sustained Reductive Removal of Uranium in Contaminated Aquifers. Final report

    International Nuclear Information System (INIS)

    Lovley, Derek R.

    2008-01-01

    The studies completed under this grant significantly advanced the understanding and design of strategies for in situ uranium bioremediation. Novel strategies identified show promise to make in situ uranium bioremediation technically simpler and less expensive. As detailed, important findings included: (1) Development of an electron donor delivery strategy to prolong the in situ activity of Geobacter species and enhance the removal of uranium from the groundwater; (2) Demonstration that reproducible year-to-year field experiments were possible at the ERSP study site in Rifle, CO, making hypothesis-driven field experimentation possible; (3) Elucidation of the geochemical and microbiological heterogeneities with the subsurface during in situ uranium bioremediation, which must be accounted for to accurately model the bioremediation process; (4) The discovery that most of the U(VI) contamination at the Rifle site is sediment-associated rather than mobile in the groundwater, as previously considered; (5) The finding that unlike soluble U(VI), sediment-associated U(VI) is not microbially reducible; (6) The demonstration that electrodes may be an effective alternative to acetate as an electron donor to promote microbial U(VI) reduction in the subsurface with the added benefit that electrode-promoted microbial U(VI) reduction offers the possibility of removing the immobilized uranium from the subsurface; and (7) The finding that, after extended acetate inputs, U(VI) continues to be removed from groundwater long after the introduction of acetate into the subsurface is terminated and that this appears to be due to adsorption onto biomass. This potentially will make in situ uranium bioremediation much less expensive than previously envisioned.

  16. Using DNA-Stable Isotope Probing to Identify MTBE- and TBA-Degrading Microorganisms in Contaminated Groundwater.

    Science.gov (United States)

    Key, Katherine C; Sublette, Kerry L; Duncan, Kathleen; Mackay, Douglas M; Scow, Kate M; Ogles, Dora

    2013-01-01

    Although the anaerobic biodegradation of methyl tert -butyl ether (MTBE) and tert -butyl alcohol (TBA) has been documented in the laboratory and the field, knowledge of the microorganisms and mechanisms involved is still lacking. In this study, DNA-stable isotope probing (SIP) was used to identify microorganisms involved in anaerobic fuel oxygenate biodegradation in a sulfate-reducing MTBE and TBA plume. Microorganisms were collected in the field using Bio-Sep® beads amended with 13 C 5 -MTBE, 13 C 1 -MTBE (only methoxy carbon labeled), or 13 C 4 -TBA. 13 C-DNA and 12 C-DNA extracted from the Bio-Sep beads were cloned and 16S rRNA gene sequences were used to identify the indigenous microorganisms involved in degrading the methoxy group of MTBE and the tert -butyl group of MTBE and TBA. Results indicated that microorganisms were actively degrading 13 C-labeled MTBE and TBA in situ and the 13 C was incorporated into their DNA. Several sequences related to known MTBE- and TBA-degraders in the Burkholderiales and the Sphingomonadales orders were detected in all three 13 C clone libraries and were likely to be primary degraders at the site. Sequences related to sulfate-reducing bacteria and iron-reducers, such as Geobacter and Geothrix , were only detected in the clone libraries where MTBE and TBA were fully labeled with 13 C, suggesting that they were involved in processing carbon from the tert -butyl group. Sequences similar to the Pseudomonas genus predominated in the clone library where only the methoxy carbon of MTBE was labeled with 13 C. It is likely that members of this genus were secondary degraders cross-feeding on 13 C-labeled metabolites such as acetate.

  17. Time-dependent bacterial community and electrochemical characterizations of cathodic biofilms in the surfactant-amended sediment-based bioelectrochemical reactor with enhanced 2,3,4,5-tetrachlorobiphenyl dechlorination.

    Science.gov (United States)

    Wan, Hui; Yi, Xiaoyun; Liu, Xiaoping; Feng, Chunhua; Dang, Zhi; Wei, Chaohai

    2018-05-01

    Applying an electric field to stimulate the microbial reductive dechlorination of polychlorinated biphenyls (PCBs) represents a promising approach for bioremediation of PCB-contaminated sites. This study aimed to demonstrate the biocathodic film-facilitated reduction of PCB 61 in a sediment-based bioelectrochemical reactor (BER) and, more importantly, the characterizations of electrode-microbe interaction from microbial and electrochemical perspectives particularly in a time-dependent manner. The application of a cathodic potential (-0.45 V vs. SHE) significantly improved the rate and extent of PCB 61 dechlorination compared to the open-circuit scenario (without electrical stimulation), and the addition of an external surfactant further increased the dechlorination, with Tween 80 exerting more pronounced effects than rhamnolipid. The bacterial composition of the biofilms and the bioelectrochemical kinetics of the BERs were found to be time-dependent and to vary considerably with the incubation time and slightly with the coexistence of an external surfactant. Excellent correlations were observed between the dechlorination rate and the relative abundance of Dehalogenimonas, Dechloromonas, and Geobacter, the dechlorination rate and the cathodic current density recorded from the chronoamperometry tests, and the dechlorination rate and the charge transfer resistance derived from the electrochemical impedance tests, with respect to the 120 day-operation. After day 120, PCB 61 was resistant to further appreciable reduction, but substantial hydrogen production was detected, and the bacterial community and electrochemical parameters observed on day 180 were not distinctly different from those on day 120. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Bicarbonate Impact on U(VI) Bioreduction in a Shallow Alluvial Aquifer

    Energy Technology Data Exchange (ETDEWEB)

    Long, Philip E.; Williams, Kenneth H.; Davis, James A.; Fox, Patricia M.; Wilkins, Michael J.; Yabusaki, Steven B.; Fang, Yilin; Waichler, Scott R.; Berman, Elena S.; Gupta, Manish; Chandler, Darrell P.; Murray, Christopher J.; Peacock, Aaron D.; Giloteaux, L.; Handley, Kim M.; Lovley, Derek R.; Banfield, Jillian F.

    2015-02-01

    Field-scale biostimulation and desorption tracer experiments conducted in a uranium (U) contaminated, shallow alluvial aquifer have provided insight into the coupling of microbiology, biogeochemistry, and hydrogeology that control U mobility in the subsurface. Initial experiments successfully tested the concept that Fe-reducing bacteria such as Geobacter sp. could enzymatically reduce soluble U(VI) to insoluble U(IV) during in situ electron donor amendment (Anderson et al. 2003, Williams et al. 2011). In parallel, in situ desorption tracer tests using bicarbonate amendment demonstrated rate-limited U(VI) desorption (Fox et al. 2012). These results and prior laboratory studies underscored the importance of enzymatic U(VI)-reduction and suggested the ability to combine desorption and bioreduction of U(VI). Here we report the results of a new field experiment in which bicarbonate-promoted uranium desorption and acetate amendment were combined and compared to an acetate amendment-only experiment in the same experimental plot. Results confirm that bicarbonate amendment to alluvial aquifer desorbs U(VI) and increases the abundance of Ca-uranyl-carbonato complexes. At the same time, that the rate of acetate-promoted enzymatic U(VI) reduction was greater in the presence of added bicarbonate in spite of the increased dominance of Ca-uranyl-carbonato aqueous complexes. A model-simulated peak rate of U(VI) reduction was ~3.8 times higher during acetate-bicarbonate treatment than under acetate-only conditions. Lack of consistent differences in microbial community structure between acetate-bicarbonate and acetate-only treatments suggest that a significantly higher rate of U(VI) reduction the bicarbonate-impacted sediment may be due to a higher intrinsic rate of microbial reduction induced by elevated concentrations of the bicarbonate oxyanion. The findings indicate that bicarbonate amendment may be useful in improving the engineered bioremediation of uranium in aquifers.

  19. Bicarbonate impact on U(VI) bioreduction in a shallow alluvial aquifer

    Science.gov (United States)

    Long, Philip E.; Williams, Kenneth H.; Davis, James A.; Fox, Patricia M.; Wilkins, Michael J.; Yabusaki, Steven B.; Fang, Yilin; Waichler, Scott R.; Berman, Elena S. F.; Gupta, Manish; Chandler, Darrell P.; Murray, Chris; Peacock, Aaron D.; Giloteaux, Ludovic; Handley, Kim M.; Lovley, Derek R.; Banfield, Jillian F.

    2015-02-01

    Field-scale biostimulation and desorption tracer experiments conducted in a uranium (U) contaminated, shallow alluvial aquifer have provided insight into the coupling of microbiology, biogeochemistry, and hydrogeology that control U mobility in the subsurface. Initial experiments successfully tested the concept that Fe-reducing bacteria such as Geobacter sp. could enzymatically reduce soluble U(VI) to insoluble U(IV) during in situ electron donor amendment (Anderson et al., 2003; Williams et al., 2011). In parallel, in situ desorption tracer tests using bicarbonate amendment demonstrated rate-limited U(VI) desorption (Fox et al., 2012). These results and prior laboratory studies underscored the importance of enzymatic U(VI)-reduction and suggested the ability to combine desorption and bioreduction of U(VI). Here we report the results of a new field experiment in which bicarbonate-promoted uranium desorption and acetate amendment were combined and compared to an acetate amendment-only experiment in the same experimental plot. Results confirm that bicarbonate amendment to alluvial aquifer sediments desorbs U(VI) and increases the abundance of Ca-uranyl-carbonato complexes. At the same time, the rate of acetate-promoted enzymatic U(VI) reduction was greater in the presence of added bicarbonate in spite of the increased dominance of Ca-uranyl-carbonato aqueous complexes. A model-simulated peak rate of U(VI) reduction was ∼3.8 times higher during acetate-bicarbonate treatment than under acetate-only conditions. Lack of consistent differences in microbial community structure between acetate-bicarbonate and acetate-only treatments suggest that a significantly higher rate of U(VI) reduction in the bicarbonate-impacted sediment may be due to a higher intrinsic rate of microbial reduction induced by elevated concentrations of the bicarbonate oxyanion. The findings indicate that bicarbonate amendment may be useful in improving the engineered bioremediation of uranium in

  20. Syntrophic interactions improve power production in formic acid fed MFCs operated with set anode potentials or fixed resistances

    KAUST Repository

    Sun, Dan; Call, Douglas F.; Kiely, Patrick D.; Wang, Aijie; Logan, Bruce E.

    2011-01-01

    Formic acid is a highly energetic electron donor but it has previously resulted in low power densities in microbial fuel cells (MFCs). Three different set anode potentials (-0.30, -0.15, and +0.15V; vs. a standard hydrogen electrode, SHE) were used to evaluate syntrophic interactions in bacterial communities for formic acid degradation relative to a non-controlled, high resistance system (1,000Ω external resistance). No current was generated at -0.30V, suggesting a lack of direct formic acid oxidation (standard reduction potential: -0.40V). More positive potentials that allowed for acetic acid utilization all produced current, with the best performance at -0.15V. The anode community in the -0.15V reactor, based on 16S rDNA clone libraries, was 58% Geobacter sulfurreducens and 17% Acetobacterium, with lower proportions of these genera found in the other two MFCs. Acetic acid was detected in all MFCs suggesting that current generation by G. sulfurreducens was dependent on acetic acid production by Acetobacterium. When all MFCs were subsequently operated at an external resistance for maximum power production (100Ω for MFCs originally set at -0.15 and +0.15V; 150Ω for the control), they produced similar power densities and exhibited the same midpoint potential of -0.15V in first derivative cyclic voltammetry scans. All of the mixed communities converged to similar proportions of the two predominant genera (ca. 52% G. sulfurreducens and 22% Acetobacterium). These results show that syntrophic interactions can be enhanced through setting certain anode potentials, and that long-term performance produces stable and convergent communities. © 2011 Wiley Periodicals, Inc.

  1. Syntrophic interactions improve power production in formic acid fed MFCs operated with set anode potentials or fixed resistances

    KAUST Repository

    Sun, Dan

    2011-10-24

    Formic acid is a highly energetic electron donor but it has previously resulted in low power densities in microbial fuel cells (MFCs). Three different set anode potentials (-0.30, -0.15, and +0.15V; vs. a standard hydrogen electrode, SHE) were used to evaluate syntrophic interactions in bacterial communities for formic acid degradation relative to a non-controlled, high resistance system (1,000Ω external resistance). No current was generated at -0.30V, suggesting a lack of direct formic acid oxidation (standard reduction potential: -0.40V). More positive potentials that allowed for acetic acid utilization all produced current, with the best performance at -0.15V. The anode community in the -0.15V reactor, based on 16S rDNA clone libraries, was 58% Geobacter sulfurreducens and 17% Acetobacterium, with lower proportions of these genera found in the other two MFCs. Acetic acid was detected in all MFCs suggesting that current generation by G. sulfurreducens was dependent on acetic acid production by Acetobacterium. When all MFCs were subsequently operated at an external resistance for maximum power production (100Ω for MFCs originally set at -0.15 and +0.15V; 150Ω for the control), they produced similar power densities and exhibited the same midpoint potential of -0.15V in first derivative cyclic voltammetry scans. All of the mixed communities converged to similar proportions of the two predominant genera (ca. 52% G. sulfurreducens and 22% Acetobacterium). These results show that syntrophic interactions can be enhanced through setting certain anode potentials, and that long-term performance produces stable and convergent communities. © 2011 Wiley Periodicals, Inc.

  2. Characterization of bacterial and archaeal communities in air-cathode microbial fuel cells, open circuit and sealed-off reactors

    KAUST Repository

    Chehab, Noura A.

    2013-06-18

    A large percentage of organic fuel consumed in a microbial fuel cell (MFC) is lost as a result of oxygen transfer through the cathode. In order to understand how this oxygen transfer affects the microbial community structure, reactors were operated in duplicate using three configurations: closed circuit (CC; with current generation), open circuit (OC; no current generation), and sealed off cathodes (SO; no current, with a solid plate placed across the cathode). Most (98 %) of the chemical oxygen demand (COD) was removed during power production in the CC reactor (maximum of 640 ± 10 mW/m 2), with a low percent of substrate converted to current (coulombic efficiency of 26.5 ± 2.1 %). Sealing the cathode reduced COD removal to 7 %, but with an open cathode, there was nearly as much COD removal by the OC reactor (94.5 %) as the CC reactor. Oxygen transfer into the reactor substantially affected the composition of the microbial communities. Based on analysis of the biofilms using 16S rRNA gene pyrosequencing, microbes most similar to Geobacter were predominant on the anodes in the CC MFC (72 % of sequences), but the most abundant bacteria were Azoarcus (42 to 47 %) in the OC reactor, and Dechloromonas (17 %) in the SO reactor. Hydrogenotrophic methanogens were most predominant, with sequences most similar to Methanobacterium in the CC and SO reactor, and Methanocorpusculum in the OC reactors. These results show that oxygen leakage through the cathode substantially alters the bacterial anode communities, and that hydrogenotrophic methanogens predominate despite high concentrations of acetate. The predominant methanogens in the CC reactor most closely resembled those in the SO reactor, demonstrating that oxygen leakage alters methanogenic as well as general bacterial communities. © 2013 Springer-Verlag Berlin Heidelberg.

  3. Multiple paths of electron flow to current in microbial electrolysis cells fed with low and high concentrations of propionate

    KAUST Repository

    Rao, Hari Ananda

    2016-03-03

    Microbial electrolysis cells (MECs) provide a viable approach for bioenergy generation from fermentable substrates such as propionate. However, the paths of electron flow during propionate oxidation in the anode of MECs are unknown. Here, the paths of electron flow involved in propionate oxidation in the anode of two-chambered MECs were examined at low (4.5 mM) and high (36 mM) propionate concentrations. Electron mass balances and microbial community analysis revealed that multiple paths of electron flow (via acetate/H2 or acetate/formate) to current could occur simultaneously during propionate oxidation regardless of the concentration tested. Current (57–96 %) was the largest electron sink and methane (0–2.3 %) production was relatively unimportant at both concentrations based on electron balances. At a low propionate concentration, reactors supplemented with 2-bromoethanesulfonate had slightly higher coulombic efficiencies than reactors lacking this methanogenesis inhibitor. However, an opposite trend was observed at high propionate concentration, where reactors supplemented with 2-bromoethanesulfonate had a lower coulombic efficiency and there was a greater percentage of electron loss (23.5 %) to undefined sinks compared to reactors without 2-bromoethanesulfonate (11.2 %). Propionate removal efficiencies were 98 % (low propionate concentration) and 78 % (high propionate concentration). Analysis of 16S rRNA gene pyrosequencing revealed the dominance of sequences most similar to Geobacter sulfurreducens PCA and G. sulfurreducens subsp. ethanolicus. Collectively, these results provide new insights on the paths of electron flow during propionate oxidation in the anode of MECs fed with low and high propionate concentrations.

  4. Isolation of phyllosilicate-iron redox cycling microorganisms from an illite-smectite rich hydromorphic soil.

    Science.gov (United States)

    Shelobolina, Evgenya; Konishi, Hiromi; Xu, Huifang; Benzine, Jason; Xiong, Mai Yia; Wu, Tao; Blöthe, Marco; Roden, Eric

    2012-01-01

    The biogeochemistry of phyllosilicate-Fe redox cycling was studied in a Phalaris arundinacea (reed canary grass) dominated redoximorphic soil from Shovelers Sink, a small glacial depression near Madison, WI. The clay size fraction of Shovelers Sink soil accounts for 16% of the dry weight of the soil, yet contributes 74% of total Fe. The dominant mineral in the clay size fraction is mixed layer illite-smectite, and in contrast to many other soils and sediments, Fe(III) oxides are present in low abundance. We examined the Fe biogeochemistry of Shovelers Sink soils, estimated the abundance of Fe redox cycling microorganisms, and isolated in pure culture representative phyllosilicate-Fe oxidizing and reducing organisms. The abundance of phyllosilicate-Fe reducing and oxidizing organisms was low compared to culturable aerobic heterotrophs. Both direct isolation and dilution-to-extinction approaches using structural Fe(II) in Bancroft biotite as a Fe(II) source, and O(2) as the electron acceptor, resulted in recovery of common rhizosphere organisms including Bradyrhizobium spp. and strains of Cupriavidus necator and Ralstonia solanacearum. In addition to oxidizing biotite and soluble Fe(II) with O(2), each of these isolates was able to oxidize Fe(II) in reduced NAu-2 smectite with [Formula: see text] as the electron acceptor. Oxidized NAu-2 smectite or amorphous Fe(III) oxide served as electron acceptors for enrichment and isolation of Fe(III)-reducing microorganisms, resulting in recovery of a strain related to Geobacter toluenoxydans. The ability of the recovered microorganisms to cycle phyllosilicate-Fe was verified in an experiment with native Shovelers Sink clay. This study confirms that Fe in the native Shovelers Sink clay is readily available for microbial redox transformation and can be cycled by the Fe(III)-reducing and Fe(II)-oxidizing microorganisms recovered from the soil.

  5. Isolation of Phyllosilicate–Iron Redox Cycling Microorganisms from an Illite–Smectite Rich Hydromorphic Soil

    Science.gov (United States)

    Shelobolina, Evgenya; Konishi, Hiromi; Xu, Huifang; Benzine, Jason; Xiong, Mai Yia; Wu, Tao; Blöthe, Marco; Roden, Eric

    2012-01-01

    The biogeochemistry of phyllosilicate–Fe redox cycling was studied in a Phalaris arundinacea (reed canary grass) dominated redoximorphic soil from Shovelers Sink, a small glacial depression near Madison, WI. The clay size fraction of Shovelers Sink soil accounts for 16% of the dry weight of the soil, yet contributes 74% of total Fe. The dominant mineral in the clay size fraction is mixed layer illite–smectite, and in contrast to many other soils and sediments, Fe(III) oxides are present in low abundance. We examined the Fe biogeochemistry of Shovelers Sink soils, estimated the abundance of Fe redox cycling microorganisms, and isolated in pure culture representative phyllosilicate–Fe oxidizing and reducing organisms. The abundance of phyllosilicate–Fe reducing and oxidizing organisms was low compared to culturable aerobic heterotrophs. Both direct isolation and dilution-to-extinction approaches using structural Fe(II) in Bancroft biotite as a Fe(II) source, and O2 as the electron acceptor, resulted in recovery of common rhizosphere organisms including Bradyrhizobium spp. and strains of Cupriavidus necator and Ralstonia solanacearum. In addition to oxidizing biotite and soluble Fe(II) with O2, each of these isolates was able to oxidize Fe(II) in reduced NAu-2 smectite with NO3- as the electron acceptor. Oxidized NAu-2 smectite or amorphous Fe(III) oxide served as electron acceptors for enrichment and isolation of Fe(III)-reducing microorganisms, resulting in recovery of a strain related to Geobacter toluenoxydans. The ability of the recovered microorganisms to cycle phyllosilicate–Fe was verified in an experiment with native Shovelers Sink clay. This study confirms that Fe in the native Shovelers Sink clay is readily available for microbial redox transformation and can be cycled by the Fe(III)-reducing and Fe(II)-oxidizing microorganisms recovered from the soil. PMID:22493596

  6. Chlorine Isotope Effects from Isotope Ratio Mass Spectrometry Suggest Intramolecular C-Cl Bond Competition in Trichloroethene (TCE Reductive Dehalogenation

    Directory of Open Access Journals (Sweden)

    Stefan Cretnik

    2014-05-01

    Full Text Available Chlorinated ethenes are prevalent groundwater contaminants. To better constrain (biochemical reaction mechanisms of reductive dechlorination, the position-specificity of reductive trichloroethene (TCE dehalogenation was investigated. Selective biotransformation reactions (i of tetrachloroethene (PCE to TCE in cultures of Desulfitobacterium sp. strain Viet1; and (ii of TCE to cis-1,2-dichloroethene (cis-DCE in cultures of Geobacter lovleyi strain SZ were investigated. Compound-average carbon isotope effects were −19.0‰ ± 0.9‰ (PCE and −12.2‰ ± 1.0‰ (TCE (95% confidence intervals. Using instrumental advances in chlorine isotope analysis by continuous flow isotope ratio mass spectrometry, compound-average chorine isotope effects were measured for PCE (−5.0‰ ± 0.1‰ and TCE (−3.6‰ ± 0.2‰. In addition, position-specific kinetic chlorine isotope effects were determined from fits of reactant and product isotope ratios. In PCE biodegradation, primary chlorine isotope effects were substantially larger (by −16.3‰ ± 1.4‰ (standard error than secondary. In TCE biodegradation, in contrast, the product cis-DCE reflected an average isotope effect of −2.4‰ ± 0.3‰ and the product chloride an isotope effect of −6.5‰ ± 2.5‰, in the original positions of TCE from which the products were formed (95% confidence intervals. A greater difference would be expected for a position-specific reaction (chloride would exclusively reflect a primary isotope effect. These results therefore suggest that both vicinal chlorine substituents of TCE were reactive (intramolecular competition. This finding puts new constraints on mechanistic scenarios and favours either nucleophilic addition by Co(I or single electron transfer as reductive dehalogenation mechanisms.

  7. Biostimulation of anaerobic BTEX biodegradation under fermentative methanogenic conditions at source-zone groundwater contaminated with a biodiesel blend (B20).

    Science.gov (United States)

    Ramos, Débora Toledo; da Silva, Márcio Luis Busi; Chiaranda, Helen Simone; Alvarez, Pedro J J; Corseuil, Henry Xavier

    2013-06-01

    Field experiments were conducted to assess the potential for anaerobic biostimulation to enhance BTEX biodegradation under fermentative methanogenic conditions in groundwater impacted by a biodiesel blend (B20, consisting of 20 % v/v biodiesel and 80 % v/v diesel). B20 (100 L) was released at each of two plots through an area of 1 m(2) that was excavated down to the water table, 1.6 m below ground surface. One release was biostimulated with ammonium acetate, which was added weekly through injection wells near the source zone over 15 months. The other release was not biostimulated and served as a baseline control simulating natural attenuation. Ammonium acetate addition stimulated the development of strongly anaerobic conditions, as indicated by near-saturation methane concentrations. BTEX removal began within 8 months in the biostimulated source zone, but not in the natural attenuation control, where BTEX concentrations were still increasing (due to source dissolution) 2 years after the release. Phylogenetic analysis using quantitative PCR indicated an increase in concentration and relative abundance of Archaea (Crenarchaeota and Euryarchaeota), Geobacteraceae (Geobacter and Pelobacter spp.) and sulfate-reducing bacteria (Desulfovibrio, Desulfomicrobium, Desulfuromusa, and Desulfuromonas) in the biostimulated plot relative to the control. Apparently, biostimulation fortuitously enhanced the growth of putative anaerobic BTEX degraders and associated commensal microorganisms that consume acetate and H2, and enhance the thermodynamic feasibility of BTEX fermentation. This is the first field study to suggest that anaerobic-methanogenic biostimulation could enhance source zone bioremediation of groundwater aquifers impacted by biodiesel blends.

  8. Intercellular wiring enables electron transfer between methanotrophic archaea and bacteria.

    Science.gov (United States)

    Wegener, Gunter; Krukenberg, Viola; Riedel, Dietmar; Tegetmeyer, Halina E; Boetius, Antje

    2015-10-22

    The anaerobic oxidation of methane (AOM) with sulfate controls the emission of the greenhouse gas methane from the ocean floor. In marine sediments, AOM is performed by dual-species consortia of anaerobic methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB) inhabiting the methane-sulfate transition zone. The biochemical pathways and biological adaptations enabling this globally relevant process are not fully understood. Here we study the syntrophic interaction in thermophilic AOM (TAOM) between ANME-1 archaea and their consortium partner SRB HotSeep-1 (ref. 6) at 60 °C to test the hypothesis of a direct interspecies exchange of electrons. The activity of TAOM consortia was compared to the first ANME-free culture of an AOM partner bacterium that grows using hydrogen as the sole electron donor. The thermophilic ANME-1 do not produce sufficient hydrogen to sustain the observed growth of the HotSeep-1 partner. Enhancing the growth of the HotSeep-1 partner by hydrogen addition represses methane oxidation and the metabolic activity of ANME-1. Further supporting the hypothesis of direct electron transfer between the partners, we observe that under TAOM conditions, both ANME and the HotSeep-1 bacteria overexpress genes for extracellular cytochrome production and form cell-to-cell connections that resemble the nanowire structures responsible for interspecies electron transfer between syntrophic consortia of Geobacter. HotSeep-1 highly expresses genes for pili production only during consortial growth using methane, and the nanowire-like structures are absent in HotSeep-1 cells isolated with hydrogen. These observations suggest that direct electron transfer is a principal mechanism in TAOM, which may also explain the enigmatic functioning and specificity of other methanotrophic ANME-SRB consortia.

  9. Diversity of Metabolically Active Bacteria in Water-Flooded High-Temperature Heavy Oil Reservoir

    Directory of Open Access Journals (Sweden)

    Tamara N. Nazina

    2017-04-01

    Full Text Available The goal of this work was to study the overall genomic diversity of microorganisms of the Dagang high-temperature oilfield (PRC and to characterize the metabolically active fraction of these populations. At this water-flooded oilfield, the microbial community of formation water from the near-bottom zone of an injection well where the most active microbial processes of oil degradation occur was investigated using molecular, cultural, radiotracer, and physicochemical techniques. The samples of microbial DNA and RNA from back-flushed water were used to obtain the clone libraries for the 16S rRNA gene and cDNA of 16S rRNA, respectively. The DNA-derived clone libraries were found to contain bacterial and archaeal 16S rRNA genes and the alkB genes encoding alkane monooxygenases similar to those encoded by alkB-geo1 and alkB-geo6 of geobacilli. The 16S rRNA genes of methanogens (Methanomethylovorans, Methanoculleus, Methanolinea, Methanothrix, and Methanocalculus were predominant in the DNA-derived library of Archaea cloned sequences; among the bacterial sequences, the 16S rRNA genes of members of the genus Geobacillus were the most numerous. The RNA-derived library contained only bacterial cDNA of the 16S rRNA sequences belonging to metabolically active aerobic organotrophic bacteria (Tepidimonas, Pseudomonas, Acinetobacter, as well as of denitrifying (Azoarcus, Tepidiphilus, Calditerrivibrio, fermenting (Bellilinea, iron-reducing (Geobacter, and sulfate- and sulfur-reducing bacteria (Desulfomicrobium, Desulfuromonas. The presence of the microorganisms of the main functional groups revealed by molecular techniques was confirmed by the results of cultural, radioisotope, and geochemical research. Functioning of the mesophilic and thermophilic branches was shown for the microbial food chain of the near-bottom zone of the injection well, which included the microorganisms of the carbon, sulfur, iron, and nitrogen cycles.

  10. Effects of 1,1,1-Trichloroethane and Triclocarban on Reductive Dechlorination of Trichloroethene in a TCE-Reducing Culture

    Directory of Open Access Journals (Sweden)

    Li-Lian Wen

    2017-08-01

    Full Text Available Chlorinated compounds were generally present in the environment due to widespread use in the industry. A short-term study was performed to evaluate the effects of 1,1,1- trichloroethane (TCA and triclocarban (TCC on trichloroethene (TCE removal in a reactor fed with lactate as the sole electron donor. Both TCA and TCC inhibited TCE reduction, but the TCC had a more pronounced effect compared to TCA. The TCE-reducing culture, which had never been exposed to TCA before, reductively dechlorinated TCA to 1,1-dichloroethane (DCA. Below 15 μM, TCA had little effect on the transformation of TCE to cis-dichloroethene (DCE; however, the reduction of cis-DCE and vinyl chloride (VC were more sensitive to TCA, and ethene production was completely inhibited when the concentration of TCA was above 15 μM. In cultures amended with TCC, the reduction of TCE was severely affected, even at concentrations as low as 0.3 μM; all the cultures stalled at VC, and no ethene was detected. The cultures that fully transformed TCE to ethene contained 5.2–8.1% Dehalococcoides. Geobacter and Desulfovibrio, the bacteria capable of partially reducing TCE to DCE, were detected in all cultures, but both represented a larger proportion of the community in TCC-amended cultures. All cultures were dominated by Clostridium_sensu_stricto_7, a genus that belongs to Firmicutes with proportions ranging from 40.9% (in a high TCC (15 μM culture to 88.2%. Methanobacteria was detected at levels of 1.1–12.7%, except in cultures added with 15 and 30 μM TCA, in which they only accounted for ∼0.4%. This study implies further environmental factors needed to be considered in the successful bioremediation of TCE in contaminated sites.

  11. Microbial electro-catalysis in fuel cell

    International Nuclear Information System (INIS)

    Dumas, Claire

    2007-01-01

    Microbial fuel cells (MFC) are devices that ensure the direct conversion of organic matter into electricity using bacterial bio-films as the catalysts of the electrochemical reactions. This study aims at improving the comprehension of the mechanisms involved in electron transfer pathways between the adhered bacteria and the electrodes. This optimization of the MFC power output could be done, for example, in exploring and characterizing various electrode materials. The electrolysis experiments carried out on Geobacter sulfurreducens deal with the microbial catalysis of the acetate oxidation, on the one hand, and the catalysis of the fumarate reduction on the other hand. On the anodic side, differences in current densities appeared on graphite, DSA R and stainless steel (8 A/m 2 , 5 A/m 2 and 0.7 A/m 2 respectively). These variations were explained more by materials roughness differences rather than their nature. Impedance spectroscopy study shows that the electro-active bio-film developed on stainless steel does not seem to modify the evolution of the stainless steel oxide layer, only the imposed potential remains determining. On the cathodic side, stainless steel sustained current densities more than twenty times higher than those obtained with graphite electrodes. The adhesion study of G. sulfurreducens on various materials in a flow cell, suggests that the bio-films resist to the hydrodynamic constraints and are not detached under a shear stress threshold value. The installation of two MFC prototypes, one in a sea station and the other directly in Genoa harbour (Italy) confirms some results obtained in laboratory and were promising for a MFC scale-up. (author) [fr

  12. Microbial Community Structure and Function Indicate the Severity of Chromium Contamination of the Yellow River

    Directory of Open Access Journals (Sweden)

    Yaxin Pei

    2018-01-01

    Full Text Available The Yellow River is the most important water resource in northern China. In the recent past, heavy metal contamination has become severe due to industrial processes and other anthropogenic activities. In this study, riparian soil samples with varying levels of chromium (Cr pollution severity were collected along the Gansu industrial reach of the Yellow River, including samples from uncontaminated sites (XC, XGU, slightly contaminated sites (LJX, XGD, and heavily contaminated sites (CG, XG. The Cr concentrations of these samples varied from 83.83 mg⋅kg-1 (XGU to 506.58 mg⋅kg-1 (XG. The chromate [Cr (VI] reducing ability in the soils collected in this study followed the sequence of the heavily contaminated > slightly contaminated > the un-contaminated. Common Cr remediation genes chrA and yieF were detected in the XG and CG samples. qRT-PCR results showed that the expression of chrA was up-regulated four and threefold in XG and CG samples, respectively, whereas the expression of yieF was up-regulated 66- and 7-fold in the same samples after 30 min treatment with Cr (VI. The copy numbers of chrA and yieF didn’t change after 35 days incubation with Cr (VI. The microbial communities in the Cr contaminated sampling sites were different from those in the uncontaminated samples. Especially, the relative abundances of Firmicutes and Bacteroidetes were higher while Actinobacteria was lower in the contaminated group than uncontaminated group. Further, potential indicator species, related to Cr such as Cr-remediation genera (Geobacter, PSB-M-3, Flavobacterium, and Methanosarcina; the Cr-sensitive genera (Skermanella, Iamia, Arthrobacter, and Candidatus Nitrososphaera were also identified. These data revealed that Cr shifted microbial composition and function. Further, Cr (VI reducing ability could be related with the expression of Cr remediation genes.

  13. Molecular evolution of the nif gene cluster carrying nifI1 and nifI2 genes in the Gram-positive phototrophic bacterium Heliobacterium chlorum.

    Science.gov (United States)

    Enkh-Amgalan, Jigjiddorj; Kawasaki, Hiroko; Seki, Tatsuji

    2006-01-01

    A major nif cluster was detected in the strictly anaerobic, Gram-positive phototrophic bacterium Heliobacterium chlorum. The cluster consisted of 11 genes arranged within a 10 kb region in the order nifI1, nifI2, nifH, nifD, nifK, nifE, nifN, nifX, fdx, nifB and nifV. The phylogenetic position of Hbt. chlorum was the same in the NifH, NifD, NifK, NifE and NifN trees; Hbt. chlorum formed a cluster with Desulfitobacterium hafniense, the closest neighbour of heliobacteria based on the 16S rRNA phylogeny, and two species of the genus Geobacter belonging to the Deltaproteobacteria. Two nifI genes, known to occur in the nif clusters of methanogenic archaea between nifH and nifD, were found upstream of the nifH gene of Hbt. chlorum. The organization of the nif operon and the phylogeny of individual and concatenated gene products showed that the Hbt. chlorum nif operon carrying nifI genes upstream of the nifH gene was an intermediate between the nif operon with nifI downstream of nifH (group II and III of the nitrogenase classification) and the nif operon lacking nifI (group I). Thus, the phylogenetic position of Hbt. chlorum nitrogenase may reflect an evolutionary stage of a divergence of the two nitrogenase groups, with group I consisting of the aerobic diazotrophs and group II consisting of strictly anaerobic prokaryotes.

  14. Evaluation of organic matter removal and electricity generation by using integrated microbial fuel cells for wastewater treatment.

    Science.gov (United States)

    Yamashita, Takahiro; Ishida, Mitsuyoshi; Ogino, Akifumi; Yokoyama, Hiroshi

    2016-01-01

    A floating all-in-one type of microbial fuel cell (Fa-MFC) that allows simple operation and installation in existing wastewater reservoirs for decomposition of organic matter was designed. A prototype cell was constructed by fixing a tubular floater to an assembly composed of a proton-exchange membrane and an air-cathode. To compare anode materials, carbon-cloth anodes or carbon-brush anodes were included in the assembly. The fabricated assemblies were floated in 1-L beakers filled with acetate medium. Both reactors removed acetate at a rate of 133-181 mg/L/d. The Fa-MFC quipped with brush anodes generated a 1.7-fold higher maximum power density (197 mW/m(2)-cathode area) than did that with cloth anodes (119 mW/m(2)-cathode area). To evaluate the performance of the Fa-MFCs on more realistic substrates, artificial wastewater, containing peptone and meat extract, was placed in a 2-L beaker, and the Fa-MFC with brush anodes was floated in the beaker. The Fa-MFC removed the chemical oxygen demand of the wastewater at a rate of 465-1029 mg/L/d, and generated a maximum power density of 152 mW/m(2)-cathode area. When the Fa-MFC was fed with actual livestock wastewater, the biological oxygen demand of the wastewater was removed at a rate of 45-119 mg/L/d, with electricity generation of 95 mW/m(2)-cathode area. Bacteria related to Geobacter sulfurreducens were predominantly detected in the anode biofilm, as deduced from the analysis of the 16S rRNA gene sequence.

  15. Enzymology of Electron Transport: Energy Generation with Geochemical Consequences

    Energy Technology Data Exchange (ETDEWEB)

    Dichristina, Thomas J.; Fredrickson, Jim K.; Zachara, John M.

    2005-12-20

    Dissimilatory metal-reducing bacteria (DMRB) are important components of the microbial community residing in redox-stratified freshwater and marine environments. DMRB occupy a central position in the biogeochemical cycles of metals, metalloids and radionuclides, and serve as catalysts for a variety of other environmentally important processes including biomineralization, biocorrosion, bioremediation and mediators of ground water quality. DMRB are presented, however, with a unique physiological challenge: they are required to respire anaerobically on terminal electron acceptors which are either highly insoluble (e.g., Fe(III)- and Mn(IV)-oxides) and reduced to soluble end-products or highly soluble (e.g., U(VI) and Tc(VII)) and reduced to insoluble end-products. To overcome physiological problems associated with metal and radionuclide solubility, DMRB are postulated to employ a variety of novel respiratory strategies not found in other gram-negative bacteria which respire on soluble electron acceptors such as O2, NO3- and SO42-. The novel respiratory strategies include (1) direct enzymatic reduction at the outer membrane, (2) electron shuttling pathways and (3) metal solubilization by exogenous or bacterially-produced organic ligands followed by reduction of soluble organic-metal compounds. The first section of this chapter highlights the latest findings on the enzymatic mechanisms of metal and radionuclide reduction by two of the most extensively studied DMRB (Geobacter and Shewanella), with particular emphasis on electron transport chain enzymology. The second section emphasizes the geochemical consequences of DMRB activity, including the direct and indirect effects on metal solubility, the reductive transformation of Fe- and Mn-containing minerals, and the biogeochemical cycling of metals at redox interfaces in chemically stratified environments.

  16. Enzymology of Electron Transport: Energy Generation with Geochemical Consequences

    International Nuclear Information System (INIS)

    Dichristina, Thomas J.; Fredrickson, Jim K.; Zachara, John M.

    2005-01-01

    Dissimilatory metal-reducing bacteria (DMRB) are important components of the microbial community residing in redox-stratified freshwater and marine environments. DMRB occupy a central position in the biogeochemical cycles of metals, metalloids and radionuclides, and serve as catalysts for a variety of other environmentally important processes including biomineralization, biocorrosion, bioremediation and mediators of ground water quality. DMRB are presented, however, with a unique physiological challenge: they are required to respire anaerobically on terminal electron acceptors which are either highly insoluble (e.g., Fe(III)- and Mn(IV)-oxides) and reduced to soluble end-products or highly soluble (e.g., U(VI) and Tc(VII)) and reduced to insoluble end-products. To overcome physiological problems associated with metal and radionuclide solubility, DMRB are postulated to employ a variety of novel respiratory strategies not found in other gram-negative bacteria which respire on soluble electron acceptors such as O2, NO3- and SO42-. The novel respiratory strategies include (1) direct enzymatic reduction at the outer membrane, (2) electron shuttling pathways and (3) metal solubilization by exogenous or bacterially-produced organic ligands followed by reduction of soluble organic-metal compounds. The first section of this chapter highlights the latest findings on the enzymatic mechanisms of metal and radionuclide reduction by two of the most extensively studied DMRB (Geobacter and Shewanella), with particular emphasis on electron transport chain enzymology. The second section emphasizes the geochemical consequences of DMRB activity, including the direct and indirect effects on metal solubility, the reductive transformation of Fe- and Mn-containing minerals, and the biogeochemical cycling of metals at redox interfaces in chemically stratified environments

  17. The genome of Pelobacter carbinolicus reveals surprising metabolic capabilities and physiological features

    Energy Technology Data Exchange (ETDEWEB)

    Aklujkar, Muktak [University of Massachusetts, Amherst; Haveman, Shelley [University of Massachusetts, Amherst; DiDonatoJr, Raymond [University of Massachusetts, Amherst; Chertkov, Olga [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Land, Miriam L [ORNL; Brown, Peter [University of Massachusetts, Amherst; Lovley, Derek [University of Massachusetts, Amherst

    2012-01-01

    Background: The bacterium Pelobacter carbinolicus is able to grow by fermentation, syntrophic hydrogen/formate transfer, or electron transfer to sulfur from short-chain alcohols, hydrogen or formate; it does not oxidize acetate and is not known to ferment any sugars or grow autotrophically. The genome of P. carbinolicus was sequenced in order to understand its metabolic capabilities and physiological features in comparison with its relatives, acetate-oxidizing Geobacter species. Results: Pathways were predicted for catabolism of known substrates: 2,3-butanediol, acetoin, glycerol, 1,2-ethanediol, ethanolamine, choline and ethanol. Multiple isozymes of 2,3-butanediol dehydrogenase, ATP synthase and [FeFe]-hydrogenase were differentiated and assigned roles according to their structural properties and genomic contexts. The absence of asparagine synthetase and the presence of a mutant tRNA for asparagine encoded among RNA-active enzymes suggest that P. carbinolicus may make asparaginyl-tRNA in a novel way. Catabolic glutamate dehydrogenases were discovered, implying that the tricarboxylic acid (TCA) cycle can function catabolically. A phosphotransferase system for uptake of sugars was discovered, along with enzymes that function in 2,3-butanediol production. Pyruvate: ferredoxin/flavodoxin oxidoreductase was identified as a potential bottleneck in both the supply of oxaloacetate for oxidation of acetate by the TCA cycle and the connection of glycolysis to production of ethanol. The P. carbinolicus genome was found to encode autotransporters and various appendages, including three proteins with similarity to the geopilin of electroconductive nanowires. Conclusions: Several surprising metabolic capabilities and physiological features were predicted from the genome of P. carbinolicus, suggesting that it is more versatile than anticipated.

  18. Fast microbial reduction of ferrihydrite colloids from a soil effluent

    Science.gov (United States)

    Fritzsche, Andreas; Bosch, Julian; Rennert, Thilo; Heister, Katja; Braunschweig, Juliane; Meckenstock, Rainer U.; Totsche, Kai U.

    2012-01-01

    Recent studies on the microbial reduction of synthetic iron oxide colloids showed their superior electron accepting property in comparison to bulk iron oxides. However, natural colloidal iron oxides differ in composition from their synthetic counterparts. Besides a potential effect of colloid size, microbial iron reduction may be accelerated by electron-shuttling dissolved organic matter (DOM) as well as slowed down by inhibitors such as arsenic. We examined the microbial reduction of OM- and arsenic-containing ferrihydrite colloids. Four effluent fractions were collected from a soil column experiment run under water-saturated conditions. Ferrihydrite colloids precipitated from the soil effluent and exhibited stable hydrodynamic diameters ranging from 281 (±146) nm in the effluent fraction that was collected first and 100 (±43) nm in a subsequently obtained effluent fraction. Aliquots of these oxic effluent fractions were added to anoxic low salt medium containing diluted suspensions of Geobacter sulfurreducens. Independent of the initial colloid size, the soil effluent ferrihydrite colloids were quickly and completely reduced. The rates of Fe2+ formation ranged between 1.9 and 3.3 fmol h-1 cell-1, and are in the range of or slightly exceeding previously reported rates of synthetic ferrihydrite colloids (1.3 fmol h-1 cell-1), but greatly exceeding previously known rates of macroaggregate-ferrihydrite reduction (0.07 fmol h-1 cell-1). The inhibition of microbial Fe(III) reduction by arsenic is unlikely or overridden by the concurrent enhancement induced by soil effluent DOM. These organic species may have increased the already high intrinsic reducibility of colloidal ferrihydrite owing to quinone-mediated electron shuttling. Additionally, OM, which is structurally associated with the soil effluent ferrihydrite colloids, may also contribute to the higher reactivity due to increasing solubility and specific surface area of ferrihydrite. In conclusion, ferrihydrite

  19. Comparative Microbiome Analysis of a Fusarium Wilt Suppressive Soil and a Fusarium Wilt Conducive Soil From the Châteaurenard Region

    Directory of Open Access Journals (Sweden)

    Katarzyna Siegel-Hertz

    2018-04-01

    Full Text Available Disease-suppressive soils are soils in which specific soil-borne plant pathogens cause only limited disease although the pathogen and susceptible host plants are both present. Suppressiveness is in most cases of microbial origin. We conducted a comparative metabarcoding analysis of the taxonomic diversity of fungal and bacterial communities from suppressive and non-suppressive (conducive soils as regards Fusarium wilts sampled from the Châteaurenard region (France. Bioassays based on Fusarium wilt of flax confirmed that disease incidence was significantly lower in the suppressive soil than in the conducive soil. Furthermore, we succeeded in partly transferring Fusarium wilt-suppressiveness to the conducive soil by mixing 10% (w/w of the suppressive soil into the conducive soil. Fungal diversity differed significantly between the suppressive and conducive soils. Among dominant fungal operational taxonomic units (OTUs affiliated to known genera, 17 OTUs were detected exclusively in the suppressive soil. These OTUs were assigned to the Acremonium, Chaetomium, Cladosporium, Clonostachys, Fusarium, Ceratobasidium, Mortierella, Penicillium, Scytalidium, and Verticillium genera. Additionally, the relative abundance of specific members of the bacterial community was significantly higher in the suppressive and mixed soils than in the conducive soil. OTUs found more abundant in Fusarium wilt-suppressive soils were affiliated to the bacterial genera Adhaeribacter, Massilia, Microvirga, Rhizobium, Rhizobacter, Arthrobacter, Amycolatopsis, Rubrobacter, Paenibacillus, Stenotrophomonas, and Geobacter. Several of the fungal and bacterial genera detected exclusively or more abundantly in the Fusarium wilt-suppressive soil included genera known for their activity against F. oxysporum. Overall, this study supports the potential role of known fungal and bacterial genera in Fusarium wilt suppressive soils from Châteaurenard and pinpoints new bacterial and fungal

  20. Effect of short-term alkaline intervention on the performance of buffer-free single-chamber microbial fuel cell.

    Science.gov (United States)

    Yang, Na; Ren, Yueping; Li, Xiufen; Wang, Xinhua

    2017-06-01

    Anolyte acidification is a drawback restricting the electricity generation performance of the buffer-free microbial fuel cells (MFC). In this paper, a small amount of alkali-treated anion exchange resin (AER) was placed in front of the anode in the KCl mediated single-chamber MFC to slowly release hydroxyl ions (OH - ) and neutralize the H + ions that are generated by the anodic reaction in two running cycles. This short-term alkaline intervention to the KCl anolyte has promoted the proliferation of electroactive Geobacter sp. and enhanced the self-buffering capacity of the KCl-AER-MFC. The pH of the KCl anolyte in the KCl-AER-MFC increased and became more stable in each running cycle compared with that of the KCl-MFC after the short-term alkaline intervention. The maximum power density (P max ) of the KCl-AER-MFC increased from 307.5mW·m -2 to 542.8mW·m -2 , slightly lower than that of the PBS-MFC (640.7mW·m -2 ). The coulombic efficiency (CE) of the KCl-AER-MFC increased from 54.1% to 61.2% which is already very close to that of the PBS-MFC (61.9%). The results in this paper indicate that short-term alkaline intervention to the anolyte is an effective strategy to further promote the performance of buffer-free MFCs. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. The Role of Soil Organic Matter, Nutrients, and Microbial Community Structure on the Performance of Microbial Fuel Cells

    Science.gov (United States)

    Rooney-Varga, J. N.; Dunaj, S. J.; Vallino, J. J.; Hines, M. E.; Gay, M.; Kobyljanec, C.

    2011-12-01

    Microbial fuel cells (MFCs) offer the potential for generating electricity, mitigating greenhouse gas emissions, and bioremediating pollutants through utilization of a plentiful, natural, and renewable resource: soil organic carbon. In the current study, we analyzed microbial community structure, MFC performance, and soil characteristics in different microhabitats (bulk soil, anode, and cathode) within MFCs constructed from agricultural or forest soils in order to determine how soil type and microbial dynamics influence MFC performance. MFCs were constructed with soils from agricultural and hardwood forest sites at Harvard Forest (Petersham, MA). The bulk soil characteristics were analyzed, including polyphenols, short chain fatty acids, total organic C and N, abiotic macronutrients, N and P mineralization rates, CO2 respiration rates, and MFC power output. Microbial community structure of the anodes, cathodes, and bulk soils was determined with molecular fingerprinting methods, which included terminal restriction length polymorphism (T-RFLP) analysis and 16S rRNA gene sequencing analysis. Our results indicated that MFCs constructed from agricultural soil had power output about 17 times that of forest soil-based MFCs and respiration rates about 10 times higher than forest soil MFCs. Agricultural soil MFCs had lower C:N ratios, polyphenol content, and acetate concentrations than forest soil MFCs, suggesting that active agricultural MFC microbial communities were supported by higher quality organic carbon. Microbial community profile data indicate that the microbial communities at the anode of the high power MFCs were less diverse than in low power MFCs and were dominated by Deltaproteobacteria, Geobacter, and, to a lesser extent, Clostridia, while low-power MFC anode communities were dominated by Clostridia. These data suggest that the presence of organic carbon substrate (acetate) was not the major limiting factor in selecting for highly electrogenic microbial

  2. Characterization of bacterial and archaeal communities in air-cathode microbial fuel cells, open circuit and sealed-off reactors

    KAUST Repository

    Chehab, Noura A.; Li, Dong; Amy, Gary L.; Logan, Bruce E.; Saikaly, Pascal

    2013-01-01

    A large percentage of organic fuel consumed in a microbial fuel cell (MFC) is lost as a result of oxygen transfer through the cathode. In order to understand how this oxygen transfer affects the microbial community structure, reactors were operated in duplicate using three configurations: closed circuit (CC; with current generation), open circuit (OC; no current generation), and sealed off cathodes (SO; no current, with a solid plate placed across the cathode). Most (98 %) of the chemical oxygen demand (COD) was removed during power production in the CC reactor (maximum of 640 ± 10 mW/m 2), with a low percent of substrate converted to current (coulombic efficiency of 26.5 ± 2.1 %). Sealing the cathode reduced COD removal to 7 %, but with an open cathode, there was nearly as much COD removal by the OC reactor (94.5 %) as the CC reactor. Oxygen transfer into the reactor substantially affected the composition of the microbial communities. Based on analysis of the biofilms using 16S rRNA gene pyrosequencing, microbes most similar to Geobacter were predominant on the anodes in the CC MFC (72 % of sequences), but the most abundant bacteria were Azoarcus (42 to 47 %) in the OC reactor, and Dechloromonas (17 %) in the SO reactor. Hydrogenotrophic methanogens were most predominant, with sequences most similar to Methanobacterium in the CC and SO reactor, and Methanocorpusculum in the OC reactors. These results show that oxygen leakage through the cathode substantially alters the bacterial anode communities, and that hydrogenotrophic methanogens predominate despite high concentrations of acetate. The predominant methanogens in the CC reactor most closely resembled those in the SO reactor, demonstrating that oxygen leakage alters methanogenic as well as general bacterial communities. © 2013 Springer-Verlag Berlin Heidelberg.

  3. Inhibition of bacterial U(VI) reduction by calcium

    International Nuclear Information System (INIS)

    Brooks, Scott C.; Fredrickson, Jim K.; Carroll, S. L.; Kennedy, David W.; Zachara, John M.; Plymale, Andrew E.; Kelly, S. D.; Kemner, K. M.; Fendorf, S.

    2003-01-01

    The rapid kinetics of bacterial U(VI) reduction and low solubility of uraninite (UO2,cr) make this process an attractive option for removing uranium from groundwater. Nevertheless, conditions that may promote or inhibit U(VI) reduction are not well-defined. Recent descriptions of Ca-UO2-CO3 complexes indicate that these species may dominate the aqueous speciation of U(VI) in many environments. We monitored the bacterial reduction of U(VI) in bicarbonate-buffered solution in the presence and absence of Ca. XAFS measurements confirmed the presence of a Ca-U(VI)-CO3 complex in the initial solutions containing calcium. Calcium, at millimolar concentrations (0.45-5 mM), caused a significant decrease in the rate and extent of bacterial U(VI) reduction. Both facultative (Shewanella putrefaciens strain CN32) and obligate (Desulfovibrio desulfuricans, Geobacter sulfurreducens) anaerobic bacteria were affected by the presence of calcium. Reduction of U(VI) ceased when the calculated system Eh re ached -0.046+/- 0.001 V, based on the Ca2UO2(CO3)(3) -- > UO2,cr couple. The results are consistent with the hypothesis that U is a less energetically favorable electron acceptor when the Ca-UO2-CO3 complexes are present. The results do not support Ca inhibition caused by direct interactions with the cells or with the electron donor as the reduction of fumarate or Tc(VII)O-4(-) under identical conditions was unaffected by the presence of Ca

  4. Isolation of microorganisms involved in reduction of crystalline iron(III) oxides in natural environments.

    Science.gov (United States)

    Hori, Tomoyuki; Aoyagi, Tomo; Itoh, Hideomi; Narihiro, Takashi; Oikawa, Azusa; Suzuki, Kiyofumi; Ogata, Atsushi; Friedrich, Michael W; Conrad, Ralf; Kamagata, Yoichi

    2015-01-01

    Reduction of crystalline Fe(III) oxides is one of the most important electron sinks for organic compound oxidation in natural environments. Yet the limited number of isolates makes it difficult to understand the physiology and ecological impact of the microorganisms involved. Here, two-stage cultivation was implemented to selectively enrich and isolate crystalline iron(III) oxide reducing microorganisms in soils and sediments. Firstly, iron reducers were enriched and other untargeted eutrophs were depleted by 2-years successive culture on a crystalline ferric iron oxide (i.e., goethite, lepidocrocite, hematite, or magnetite) as electron acceptor. Fifty-eight out of 136 incubation conditions allowed the continued existence of microorganisms as confirmed by PCR amplification. High-throughput Illumina sequencing and clone library analysis based on 16S rRNA genes revealed that the enrichment cultures on each of the ferric iron oxides contained bacteria belonging to the Deltaproteobacteria (mainly Geobacteraceae), followed by Firmicutes and Chloroflexi, which also comprised most of the operational taxonomic units (OTUs) identified. Venn diagrams indicated that the core OTUs enriched with all of the iron oxides were dominant in the Geobacteraceae while each type of iron oxides supplemented selectively enriched specific OTUs in the other phylogenetic groups. Secondly, 38 enrichment cultures including novel microorganisms were transferred to soluble-iron(III) containing media in order to stimulate the proliferation of the enriched iron reducers. Through extinction dilution-culture and single colony isolation, six strains within the Deltaproteobacteria were finally obtained; five strains belonged to the genus Geobacter and one strain to Pelobacter. The 16S rRNA genes of these isolates were 94.8-98.1% identical in sequence to cultured relatives. All the isolates were able to grow on acetate and ferric iron but their physiological characteristics differed considerably in

  5. In situ bioreduction of technetium and uranium in a nitrate-contaminated aquifer

    International Nuclear Information System (INIS)

    IstokD, Jonathan; Senko, J.M.; Krumholz, Lee R.; Watson, David B.; Bogle, Mary Anna; Peacock, Aaron D.; Change, Y.J.; White, David C.

    2004-01-01

    The potential to stimulate an indigenous microbial community to reduce a mixture of U(VI) and Tc(VII) in the presence of high (120 mM) initial NO 3 - co-contamination was evaluated in a shallow unconfined aquifer using a series of single-well, push-pull tests. In the absence of added electron donor, NO 3 - , Tc(VII), and U(VI) reduction was not detectable. However, in the presence of added ethanol, glucose, or acetate to serve as electron donor, rapid NO 3 - utilization was observed. The accumulation of NO 2 - , the absence of detectable NH 4 + accumulation, and the production of N 2 O during in situ acetylene-block experiments suggest that NO 3 - was being consumed via denitrification. Tc(VII) reduction occurred concurrently with NO 3 - reduction, but U(VI) reduction was not observed until two or more donor additions resulted in iron-reducing conditions, as detected by the production of Fe(II). Reoxidation/remobilization of U(IV) was also observed in tests conducted with high (120 mM) but not low (1 mM) initial NO 3 - concentrations and not during acetylene-block experiments conducted with high initial NO 3 - . These results suggest that NO 3 - -dependent microbial U(IV) oxidation may inhibit or reverse U(VI) reduction and decrease the stability of U(IV) in this environment. Changes in viable biomass, community composition, metabolic status, and respiratory state of organisms harvested from down-well microbial samplers deployed during these tests were consistent with the conclusions that electron donor additions resulted in microbial growth, the creation of anaerobic conditions, and an increase in activity of metal-reducing organisms (e.g., Geobacter). The results demonstrate that it is possible to stimulate the simultaneous bioreduction of U(VI) and Tc(VII) mixtures commonly found with NO 3 - co-contamination at radioactive waste sites.

  6. Distribution of Anaerobic Hydrocarbon-Degrading Bacteria in Soils from King George Island, Maritime Antarctica.

    Science.gov (United States)

    Sampaio, Dayanna Souza; Almeida, Juliana Rodrigues Barboza; de Jesus, Hugo E; Rosado, Alexandre S; Seldin, Lucy; Jurelevicius, Diogo

    2017-11-01

    Anaerobic diesel fuel Arctic (DFA) degradation has already been demonstrated in Antarctic soils. However, studies comparing the distribution of anaerobic bacterial groups and of anaerobic hydrocarbon-degrading bacteria in Antarctic soils containing different concentrations of DFA are scarce. In this study, functional genes were used to study the diversity and distribution of anaerobic hydrocarbon-degrading bacteria (bamA, assA, and bssA) and of sulfate-reducing bacteria (SRB-apsR) in highly, intermediate, and non-DFA-contaminated soils collected during the summers of 2009, 2010, and 2011 from King George Island, Antarctica. Signatures of bamA genes were detected in all soils analyzed, whereas bssA and assA were found in only 4 of 10 soils. The concentration of DFA was the main factor influencing the distribution of bamA-containing bacteria and of SRB in the analyzed soils, as shown by PCR-DGGE results. bamA sequences related to genes previously described in Desulfuromonas, Lautropia, Magnetospirillum, Sulfuritalea, Rhodovolum, Rhodomicrobium, Azoarcus, Geobacter, Ramlibacter, and Gemmatimonas genera were dominant in King George Island soils. Although DFA modulated the distribution of bamA-hosting bacteria, DFA concentration was not related to bamA abundance in the soils studied here. This result suggests that King George Island soils show functional redundancy for aromatic hydrocarbon degradation. The results obtained in this study support the hypothesis that specialized anaerobic hydrocarbon-degrading bacteria have been selected by hydrocarbon concentrations present in King George Island soils.

  7. Light-dependent electrogenic activity of cyanobacteria.

    Directory of Open Access Journals (Sweden)

    John M Pisciotta

    2010-05-01

    Full Text Available Cyanobacteria account for 20-30% of Earth's primary photosynthetic productivity and convert solar energy into biomass-stored chemical energy at the rate of approximately 450 TW [1]. These single-cell microorganisms are resilient predecessors of all higher oxygenic phototrophs and can be found in self-sustaining, nitrogen-fixing communities the world over, from Antarctic glaciers to the Sahara desert [2].Here we show that diverse genera of cyanobacteria including biofilm-forming and pelagic strains have a conserved light-dependent electrogenic activity, i.e. the ability to transfer electrons to their surroundings in response to illumination. Naturally-growing biofilm-forming photosynthetic consortia also displayed light-dependent electrogenic activity, demonstrating that this phenomenon is not limited to individual cultures. Treatment with site-specific inhibitors revealed the electrons originate at the photosynthetic electron transfer chain (P-ETC. Moreover, electrogenic activity was observed upon illumination only with blue or red but not green light confirming that P-ETC is the source of electrons. The yield of electrons harvested by extracellular electron acceptor to photons available for photosynthesis ranged from 0.05% to 0.3%, although the efficiency of electron harvesting likely varies depending on terminal electron acceptor.The current study illustrates that cyanobacterial electrogenic activity is an important microbiological conduit of solar energy into the biosphere. The mechanism responsible for electrogenic activity in cyanobacteria appears to be fundamentally different from the one exploited in previously discovered electrogenic bacteria, such as Geobacter, where electrons are derived from oxidation of organic compounds and transported via a respiratory electron transfer chain (R-ETC [3], [4]. The electrogenic pathway of cyanobacteria might be exploited to develop light-sensitive devices or future technologies that convert solar

  8. Mercury methylation coupled to iron reduction by dissimilatory iron-reducing bacteria.

    Science.gov (United States)

    Si, Youbin; Zou, Yan; Liu, Xiaohong; Si, Xiongyuan; Mao, Jingdong

    2015-03-01

    Iron reduction and mercury methylation by dissimilatory iron-reducing bacteria (DIRB), Geobacter sulfurreducens and Shewanella oneidensis, were studied, and the relationship of mercury methylation coupled to iron reduction was determined. The ability of both bacteria for reducing iron was tested, and Fe(III) reduction occurred with the highest rate when ferric oxyhydroxide was used as a terminal electron acceptor. G. sulfurreducens had proven to mediate the production of methylmercury (MeHg), and a notable increase of MeHg following the addition of inorganic Hg was observed. When the initial concentration of HgCl2 was 500nM, about 177.03nM of MeHg was determined at 8d after G. sulfurreducens inoculation. S. oneidensis was tested negligible for Hg methylation and only 12.06nM of MeHg was determined. Iron reduction could potentially influence Hg methylation rates. The increase in MeHg was consistent with high rate of iron reduction, indicating that Fe(III) reduction stimulated the formation of MeHg. Furthermore, the net MeHg concentration increased at low Fe(III) additions from 1.78 to 3.57mM, and then decreased when the added Fe(III) was high from 7.14 to 17.85mM. The mercury methylation rate was suppressed with high Fe(III) additions, which might have been attributable to mercury complexation and low availability. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Optimizing Cr(VI) and Tc(VII) remediation through nano-scale biomineral engineering

    Energy Technology Data Exchange (ETDEWEB)

    Cutting, R. S.; Coker, V. S.; Telling, N. D.; Kimber, R. L.; Pearce, C. I.; Ellis, B.; Lawson, R; van der Laan, G.; Pattrick, R.A.D.; Vaughan, D.J.; Arenholz, E.; Lloyd, J. R.

    2009-09-09

    To optimize the production of biomagnetite for the bioremediation of metal oxyanion contaminated waters, the reduction of aqueous Cr(VI) to Cr(III) by two biogenic magnetites and a synthetic magnetite was evaluated under batch and continuous flow conditions. Results indicate that nano-scale biogenic magnetite produced by incubating synthetic schwertmannite powder in cell suspensions of Geobacter sulfurreducens is more efficient at reducing Cr(VI) than either biogenic nano-magnetite produced from a suspension of ferrihydrite 'gel' or synthetic nano-scale Fe{sub 3}O{sub 4} powder. Although X-ray Photoelectron Spectroscopy (XPS) measurements obtained from post-exposure magnetite samples reveal that both Cr(III) and Cr(VI) are associated with nanoparticle surfaces, X-ray Magnetic Circular Dichroism (XMCD) studies indicate that some Cr(III) has replaced octahedrally coordinated Fe in the lattice of the magnetite. Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES) measurements of total aqueous Cr in the associated solution phase indicated that, although the majority of Cr(III) was incorporated within or adsorbed to the magnetite samples, a proportion ({approx}10-15 %) was released back into solution. Studies of Tc(VII) uptake by magnetites produced via the different synthesis routes also revealed significant differences between them as regards effectiveness for remediation. In addition, column studies using a {gamma}-camera to obtain real time images of a {sup 99m}Tc(VII) radiotracer were performed to visualize directly the relative performances of the magnetite sorbents against ultra-trace concentrations of metal oxyanion contaminants. Again, the magnetite produced from schwertmannite proved capable of retaining more ({approx}20%) {sup 99m}Tc(VII) than the magnetite produced from ferrihydrite, confirming that biomagnetite production for efficient environmental remediation can be fine-tuned through careful selection of the initial Fe(III) mineral

  10. Fe-phyllosilicate redox cycling organisms from a redox transition zone in Hanford 300 Area sediments

    Directory of Open Access Journals (Sweden)

    Jason eBenzine

    2013-12-01

    Full Text Available Microorganisms capable of reducing or oxidizing structural iron (Fe in Fe-bearing phyllosilicate minerals were enriched and isolated from a subsurface redox transition zone at the Hanford 300 Area site in eastern Washington, USA. Both conventional and in situ i-chip enrichment strategies were employed. One Fe(III-reducing Geobacter (G. bremensis strain R1, Deltaproteobacteria and six Fe(II phyllosilicate-oxidizing isolates from the Alphaproteobacteria (Bradyrhizobium japonicum strains 22, is5, and in8p8, Betaproteobacteria (Cupriavidus necator strain A5-1, Dechloromonas agitata strain is5, and Actinobacteria (Nocardioides sp. strain in31 were recovered. The G. bremensis isolate grew by oxidizing acetate with the oxidized form of NAu-2 smectite as the electron acceptor. The Fe(II-oxidizers grew by oxidation of chemically reduced smectite as the energy source with nitrate as the electron acceptor. The Bradyrhizobium isolates could also carry out aerobic oxidation of biotite. This is the first report of the recovery of a Fe(II-oxidizing Nocardioides, and to date only one other Fe(II-oxidizing Bradyrhizobium is known. The 16S rRNA gene sequences of the isolates were similar to ones found in clone libraries from Hanford 300 sediments and groundwater, suggesting that such organisms may be present and active in situ. Whole genome sequencing of the isolates is underway, the results of which will enable comparative genomic analysis of mechanisms of extracellular phyllosilicate Fe redox metabolism, and facilitate development of techniques to detect the presence and expression of genes associated with microbial phyllosilicate Fe redox cycling in sediments.

  11. Similar diversity of Alphaproteobacteria and nitrogenase gene amplicons on two related Sphagnum mosses

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    Anastasia eBragina

    2012-01-01

    Full Text Available Sphagnum mosses represent a main component in ombrotrophic wetlands. They harbor a specific and diverse microbial community with essential functions for the host. To understand extend and degree of host specificity, Sphagnum fallax and S. angustifolium, two phylogenetically closely related species, which show distinct habitat preference with respect to the nutrient level, were analyzed by a multifaceted approach. Microbial fingerprints obtained by PCR-SSCP (single-strand conformation polymorphism using universal, group-specific and functional primers were highly similar. Similarity was confirmed for colonization patterns obtained by fluorescence in situ hybridization (FISH coupled with confocal laser scanning microscopy (CLSM: Alphaproteobacteria were the main colonizers inside the hyaline cells of Sphagnum leaves. A deeper survey of Alphaproteobacteria by 16S rRNA gene amplicon sequencing reveals a high diversity with Acidocella, Acidisphaera, Rhodopila and Phenylobacterium as major genera for both mosses. Pathogen defense and nitrogen fixation are important functions of Sphagnum-associated bacteria, which are fulfilled by microbial communities of both Sphagna in a similar way. NifH libraries of Sphagnum-associated microbial communities were characterized by high diversity and abundance of Alphaproteobacteria but contained also diverse amplicons of other taxa, e.g. Cyanobacteria, Geobacter and Spirochaeta. Statistically significant differences between the microbial communities of both Sphagnum species could not be discovered in any of the experimental approach. Our results show that the same close relationship, which exists between the physical, morphological and chemical characteristics of Sphagnum mosses and the ecology and function of bog ecosystems, also connects moss plantlets with their associated bacterial communities.

  12. Enhanced anaerobic dechlorination of polychlorinated biphenyl in sediments by bioanode stimulation

    International Nuclear Information System (INIS)

    Yu, Hui; Feng, Chunhua; Liu, Xiaoping; Yi, Xiaoyun; Ren, Yuan; Wei, Chaohai

    2016-01-01

    The application of a low-voltage electric field as an electron donor or acceptor to promote the bioremediation of chlorinated organic compounds represents a promising technology meeting the demand of developing an efficient and cost-effective strategy for in situ treatment of PCB-contaminated sediments. Here, we reported that bioanode stimulation with an anodic potential markedly enhanced dechlorination of 2,3,4,5-tetrachlorobiphenyl (PCB 61) contained in the sediment at an electronic waste recycling site of Qingyuan, Guangdong, China. The 110-day incubation of the bioanode with a potential poised at 0.2 V relative to saturated calomel electrode enabled 58% transformation of the total PCB 61 at the initial concentration of 100 μmol kg"−"1, while only 23% was reduced in the open-circuit reference experiment. The introduction of acetate to the bioelectrochemical reactor (BER) further improved PCB 61 transformation to 82%. Analysis of the bacterial composition showed significant community shifts in response to variations in treatment. At phylum level, the bioanode stimulation resulted in substantially increased abundance of Actinobacteria, Bacteroidetes, and Chloroflexi either capable of PCB dechlorination, or detected in the PCB-contaminated environment. At genus level, the BER contained two types of microorganisms: electrochemically active bacteria (EAB) represented by Geobacter, Ignavibacterium, and Dysgonomonas, and dechlorinating bacteria including Hydrogenophaga, Alcanivorax, Sedimentibacter, Dehalogenimonas, Comamonas and Vibrio. These results suggest that the presence of EAB can promote the population of dechlorinating bacteria which are responsible for PCB 61 transformation. - Highlights: • A bioelectrochemical reactor (BER) was constructed for anaerobic PCB dechlorination. • Bioanode stimulation substantially enhanced dechlorination of PCB 61. • Electrochemically active bacteria and dechlorinating bacteria coexisted in the BER. - Bioanode

  13. Covariance of bacterioplankton composition and environmental variables in a temperate delta system

    Science.gov (United States)

    Stepanauskas, R.; Moran, M.A.; Bergamaschi, B.A.; Hollibaugh, J.T.

    2003-01-01

    We examined seasonal and spatial variation in bacterioplankton composition in the Sacramento-San Joaquin River Delta (CA) using terminal restriction fragment length polymorphism (T-RFLP) analysis. Cloned 16S rRNA genes from this system were used for putative identification of taxa dominating the T-RFLP profiles. Both cloning and T-RFLP analysis indicated that Actinobacteria, Verrucomicrobia, Cytophaga-Flavobacterium and Proteobacteria were the most abundant bacterioplankton groups in the Delta. Despite the broad variety of sampled habitats (deep water channels, lakes, marshes, agricultural drains, freshwater and brackish areas), and the spatial and temporal differences in hydrology, temperature and water chemistry among the sampling campaigns, T-RFLP electropherograms from all samples were similar, indicating that the same bacterioplankton phylotypes dominated in the various habitats of the Delta throughout the year. However, principal component analysis (PCA) and partial least-squares regression (PLS) of T-RFLP profiles revealed consistent grouping of samples on a seasonal, but not a spatial, basis. ??-Proteobacteria related to Ralstonia, Actinobacteria related to Microthrix, and ??-Proteobacteria identical to the environmental Clone LD12 had the highest relative abundance in summer/fall T-RFLP profiles and were associated with low river flow, high pH, and a number of optical and chemical characteristics of dissolved organic carbon (DOC) indicative of an increased proportion of phytoplankton-produced organic material as opposed to allochthonous, terrestrially derived organic material. On the other hand, Geobacter-related ??-Proteobacteria showed a relative increase in abundance in T-RFLP analysis during winter/spring, and probably were washed out from watershed soils or sediment. Various phylotypes associated with the same phylogenetic division, based on tentative identification of T-RFLP fragments, exhibited diverse seasonal patterns, suggesting that ecological

  14. Teste triplo biofísico: um novo parâmetro ultra-sonográfico para prognóstico em gestações iniciais Biophysical triple test: a new ultrasonographic parameter for prognostic evaluation in early pregnancies

    Directory of Open Access Journals (Sweden)

    Joaquim R. M. Coelho

    1999-01-01

    Full Text Available Objetivos: atrasos no desenvolvimento do saco gestacional (SG, no tamanho do botão embrionário (CCN, assim como freqüências cardíacas embrionárias (FCE baixas podem ser considerados fatores de mau prognóstico na evolução da gestação. O objetivo deste trabalho foi avaliar a utilização destes 3 parâmetros em conjunto, o que denominamos Teste Triplo Biofísico (TTB. Metodo: foram avaliadas 35 gestações únicas provenientes de fertilização assistida por injeção intracitoplasmática de espermatozóide (ICSI. Todos os exames de ultra-som foram realizados por um único examinador, após 4-5 semanas da transferência de embriões (6-7 semanas de gestação, com equipamento modelo Synergy da Diasonics, sonda transvaginal de 7,0 MHz. O SG foi medido em seu maior diâmetro transverso, o CCN foi medido no sentido sagital e a FCE pelo modo B-M e Doppler. Na análise estatística foi utilizado o teste de Fisher. Resultados: considerou-se como parâmetros alterados : SG Purpose: it has been demonstrated that delays in gestational sac development, in crown-rump length and low embryonic heart rate could be indicators of poor pregnancy outcome. The purpose was to evaluate the use of these three parameters together, which we named Biophysical Triple Test (BTT. Method: thirty-five singleton pregnancies following IVF treatment with intracytoplasmatic sperm injection (ICSI were studied. All ultrasonographic scans were performed by the same examiner, 4-5 weeks after embryo transference (6-7 week gestation, using a Diasonics equipment, model Synergy, with a 7.0-MHz transvaginal transducer. The gestational sac (GS was measured at the longest transverse diameter; the crown-rump length (CRL was measured at the sagittal plan and the embryonic heart rate (EHR was calculated from frozen B and M-mode and Doppler. The statistical test used was Fisher's test. Results: the following parameters were considered abnormal: GS < 15.4 mm, CRL < 3.9 mm, EHR < 100

  15. Novel Insights Into Microbial Uranium Reduction and Immobilization

    Science.gov (United States)

    Loeffler, F. E.; Fletcher, K.; Thomas, S.; Kemner, K. M.; Boyanov, M.; Sanford, R.

    2010-12-01

    Many ferric iron- and manganese oxide-reducing bacteria affect the oxidation state and complexation of toxic radionuclides in subsurface environments. Relevant to uranium (U) speciation are bacteria that reduce predominantly water-soluble and mobile U(VI) to U(IV), which has reduced solubility and typically forms the uraninite (UO2) mineral. Gram-negative model organisms including Shewanella spp., Geobacter spp., and more recently Anaeromyxobacter spp. use U(VI) as growth-supporting electron acceptor; however, the biomass yields are lower than predicted based on the theoretical free energy changes associated with U(VI)-to-U(IV) reduction. Recent findings demonstrated that U(VI) reduction is not limited to Gram-negative bacteria, and members of the genus Desulfitobacterium, which are commonly found in soil and subsurface environments, share the ability to reduce U(VI). Interestingly, extended X-ray absorption fine structure (EXAFS) analysis demonstrated that the U(IV) produced in cultures of five Desulfitobacterium spp. was not UO2 but rather a phase or mineral composed of mononuclear U(IV) atoms. Since the properties of the reduced product influence U(IV) fate, knowledge of the diversity of U reduction mechanisms and the stability of the end products is desirable for controlling and predicting U fate. For example, UO2 is susceptible to reoxidation by oxidants, and oxic/anoxic interface processes are controlling the stability of the precipitated material. In other words, metal reducers that thrive at the oxic/anoxic interface are likely key players affecting long-term U fate. Anaeromyxobacter spp. are facultative microaerophiles and grow with oxygen as electron acceptor at partial pressures equal to or below 0.18 atm. Thus, Anaeromyxobacter are uniquely adapted to life at the oxic-anoxic interface where they consume oxygen and take advantage of oxidized metal species including U(VI) as electron acceptors. The application of 16S rRNA gene-targeted qPCR approaches

  16. Graphite anode surface modification with controlled reduction of specific aryl diazonium salts for improved microbial fuel cells power output.

    Science.gov (United States)

    Picot, Matthieu; Lapinsonnière, Laure; Rothballer, Michael; Barrière, Frédéric

    2011-10-15

    Graphite electrodes were modified with reduction of aryl diazonium salts and implemented as anodes in microbial fuel cells. First, reduction of 4-aminophenyl diazonium is considered using increased coulombic charge density from 16.5 to 200 mC/cm(2). This procedure introduced aryl amine functionalities at the surface which are neutral at neutral pH. These electrodes were implemented as anodes in "H" type microbial fuel cells inoculated with waste water, acetate as the substrate and using ferricyanide reduction at the cathode and a 1000 Ω external resistance. When the microbial anode had developed, the performances of the microbial fuel cells were measured under acetate saturation conditions and compared with those of control microbial fuel cells having an unmodified graphite anode. We found that the maximum power density of microbial fuel cell first increased as a function of the extent of modification, reaching an optimum after which it decreased for higher degree of surface modification, becoming even less performing than the control microbial fuel cell. Then, the effect of the introduction of charged groups at the surface was investigated at a low degree of surface modification. It was found that negatively charged groups at the surface (carboxylate) decreased microbial fuel cell power output while the introduction of positively charged groups doubled the power output. Scanning electron microscopy revealed that the microbial anode modified with positively charged groups was covered by a dense and homogeneous biofilm. Fluorescence in situ hybridization analyses showed that this biofilm consisted to a large extent of bacteria from the known electroactive Geobacter genus. In summary, the extent of modification of the anode was found to be critical for the microbial fuel cell performance. The nature of the chemical group introduced at the electrode surface was also found to significantly affect the performance of the microbial fuel cells. The method used for

  17. Methane-Fueled Syntrophy through Extracellular Electron Transfer: Uncovering the Genomic Traits Conserved within Diverse Bacterial Partners of Anaerobic Methanotrophic Archaea.

    Science.gov (United States)

    Skennerton, Connor T; Chourey, Karuna; Iyer, Ramsunder; Hettich, Robert L; Tyson, Gene W; Orphan, Victoria J

    2017-08-01

    The anaerobic oxidation of methane by anaerobic methanotrophic (ANME) archaea in syntrophic partnership with deltaproteobacterial sulfate-reducing bacteria (SRB) is the primary mechanism for methane removal in ocean sediments. The mechanism of their syntrophy has been the subject of much research as traditional intermediate compounds, such as hydrogen and formate, failed to decouple the partners. Recent findings have indicated the potential for extracellular electron transfer from ANME archaea to SRB, though it is unclear how extracellular electrons are integrated into the metabolism of the SRB partner. We used metagenomics to reconstruct eight genomes from the globally distributed SEEP-SRB1 clade of ANME partner bacteria to determine what genomic features are required for syntrophy. The SEEP-SRB1 genomes contain large multiheme cytochromes that were not found in previously described free-living SRB and also lack periplasmic hydrogenases that may prevent an independent lifestyle without an extracellular source of electrons from ANME archaea. Metaproteomics revealed the expression of these cytochromes at in situ methane seep sediments from three sites along the Pacific coast of the United States. Phylogenetic analysis showed that these cytochromes appear to have been horizontally transferred from metal-respiring members of the Deltaproteobacteria such as Geobacter and may allow these syntrophic SRB to accept extracellular electrons in place of other chemical/organic electron donors. IMPORTANCE Some archaea, known as anaerobic methanotrophs, are capable of converting methane into carbon dioxide when they are growing syntopically with sulfate-reducing bacteria. This partnership is the primary mechanism for methane removal in ocean sediments; however, there is still much to learn about how this syntrophy works. Previous studies have failed to identify the metabolic intermediate, such as hydrogen or formate, that is passed between partners. However, recent analysis of

  18. Microbial Dynamics During a Temporal Sequence of Bioreduction Stimulated by Emulsified Vegetable Oil

    Science.gov (United States)

    Schadt, C. W.; Gihring, T. M.; Yang, Z.; Wu, W.; Green, S.; Overholt, W.; Zhang, G.; Brandt, C. C.; Campbell, J. H.; Carroll, S. C.; Criddle, C.; Jardine, P. M.; Lowe, K.; Mehlhorn, T.; Kostka, J. E.; Watson, D. B.; Brooks, S. C.

    2011-12-01

    microbial community. Bacterial richness rebounded after nine months, although community composition remained distinct from the pre-amendment conditions. Subsequent to the experiment we have isolated several of these organisms into pure culture including representatives of probable new species of Geobacter, Desulforegula and Desulfovibrio. A hypothesized model for the functioning of these limited communities will be verified in the laboratory using defined combinations of isolates from the field where possible. These results demonstrated EVO serves as an effective electron donor source for in situ U(VI) bioreduction, and subsurface EVO degradation and metal reduction was likely mediated by successive identifiable guilds of organisms.

  19. Persistence of uranium groundwater plumes: Contrasting mechanisms at two DOE sites in the groundwater-river interaction zone

    Science.gov (United States)

    Zachara, John M.; Long, Philip E.; Bargar, John; Davis, James A.; Fox, Patricia; Fredrickson, Jim K.; Freshley, Mark D.; Konopka, Allan E.; Liu, Chongxuan; McKinley, James P.; Rockhold, Mark L.; Williams, Kenneth H.; Yabusaki, Steve B.

    2013-04-01

    We examine subsurface uranium (U) plumes at two U.S. Department of Energy sites that are located near large river systems and are influenced by groundwater-river hydrologic interaction. Following surface excavation of contaminated materials, both sites were projected to naturally flush remnant uranium contamination to levels below regulatory limits (e.g., 30 μg/L or 0.126 μmol/L; U.S. EPA drinking water standard), with 10 years projected for the Hanford 300 Area (Columbia River) and 12 years for the Rifle site (Colorado River). The rate of observed uranium decrease was much lower than expected at both sites. While uncertainty remains, a comparison of current understanding suggests that the two sites have common, but also different mechanisms controlling plume persistence. At the Hanford 300 A, the persistent source is adsorbed U(VI) in the vadose zone that is released to the aquifer during spring water table excursions. The release of U(VI) from the vadose zone and its transport within the oxic, coarse-textured aquifer sediments is dominated by kinetically-limited surface complexation. Modeling implies that annual plume discharge volumes to the Columbia River are small (oxidation of naturally reduced, contaminant U(IV) in the saturated zone and a continuous influx of U(VI) from natural, up-gradient sources influence plume persistence. Rate-limited mass transfer and surface complexation also control U(VI) migration velocity in the sub-oxic Rifle groundwater. Flux of U(VI) from the vadose zone at the Rifle site may be locally important, but it is not the dominant process that sustains the plume. A wide range in microbiologic functional diversity exists at both sites. Strains of Geobacter and other metal reducing bacteria are present at low natural abundance that are capable of enzymatic U(VI) reduction in localized zones of accumulated detrital organic carbon or after organic carbon amendment. Major differences between the sites include the geochemical nature of

  20. Persistence of uranium groundwater plumes: contrasting mechanisms at two DOE sites in the groundwater-river interaction zone.

    Science.gov (United States)

    Zachara, John M; Long, Philip E; Bargar, John; Davis, James A; Fox, Patricia; Fredrickson, Jim K; Freshley, Mark D; Konopka, Allan E; Liu, Chongxuan; McKinley, James P; Rockhold, Mark L; Williams, Kenneth H; Yabusaki, Steve B

    2013-04-01

    We examine subsurface uranium (U) plumes at two U.S. Department of Energy sites that are located near large river systems and are influenced by groundwater-river hydrologic interaction. Following surface excavation of contaminated materials, both sites were projected to naturally flush remnant uranium contamination to levels below regulatory limits (e.g., 30 μg/L or 0.126 μmol/L; U.S. EPA drinking water standard), with 10 years projected for the Hanford 300 Area (Columbia River) and 12 years for the Rifle site (Colorado River). The rate of observed uranium decrease was much lower than expected at both sites. While uncertainty remains, a comparison of current understanding suggests that the two sites have common, but also different mechanisms controlling plume persistence. At the Hanford 300 A, the persistent source is adsorbed U(VI) in the vadose zone that is released to the aquifer during spring water table excursions. The release of U(VI) from the vadose zone and its transport within the oxic, coarse-textured aquifer sediments is dominated by kinetically-limited surface complexation. Modeling implies that annual plume discharge volumes to the Columbia River are small (oxidation of naturally reduced, contaminant U(IV) in the saturated zone and a continuous influx of U(VI) from natural, up-gradient sources influence plume persistence. Rate-limited mass transfer and surface complexation also control U(VI) migration velocity in the sub-oxic Rifle groundwater. Flux of U(VI) from the vadose zone at the Rifle site may be locally important, but it is not the dominant process that sustains the plume. A wide range in microbiologic functional diversity exists at both sites. Strains of Geobacter and other metal reducing bacteria are present at low natural abundance that are capable of enzymatic U(VI) reduction in localized zones of accumulated detrital organic carbon or after organic carbon amendment. Major differences between the sites include the geochemical nature of

  1. Identification of a c-Type Cytochrome Specific for Manganese Dioxide (MnO2) Reduction in Anaeromyxobacter dehalogenans Strain 2CP-C

    Science.gov (United States)

    Pfiffner, S. M.; Nissen, S.; Liu, X.; Chourey, K.; Vishnivetskaya, T. A.; Hettich, R.; Loeffler, F.

    2014-12-01

    Anaeromyxobacter dehalogenans is a metabolically versatile Deltaproteobacterium and conserves energy from the reduction of various electron acceptors, including insoluble MnO2 and ferric oxides/oxyhydroxides (FeOOH). The goal of this study was to identify c-type cytochromes involved in electron transfer to MnO2. The characterization of deletion mutants has revealed a number of c-type cytochromes involved in electron transfer to solid metal oxides in Shewanella spp. and Geobacter spp; however, a genetic system for Anaeromyxobacter is not available. The A. dehalogenans str. 2CP-C genome encodes 68 putative c-type cytochromes, which all lack functional assignments. To identify c-type cytochromes involved in electron transfer to solid MnO2, protein expression profiles of A. dehalogenans str. 2CP-C cells grown with acetate as electron donor and MnO2, ferric citrate, FeOOH, nitrate or fumarate as electron acceptors were compared. Whole cell proteomes were analyzed after trypsin proteolysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Distinct c-type cytochrome expression patterns were observed with cells grown with different electron acceptors. A. dehalogenans str. 2CP-C grown with MnO2 expressed 25 out of the 68 c-type cytochromes encoded on the genome. The c-type cytochrome Adeh_1278 was only expressed in strain 2CP-C grown with MnO2. Reverse transcription PCR confirmed that the Adeh_1278 gene was transcribed in MnO2-grown cells but not in cells grown with other terminal electron acceptors. The expression of the Adeh_1278 gene correlated with Mn(IV) reduction activity. Adeh_1278 has three heme binding motifs and is predicted to be located in the periplasm. The identification of Adeh_1278 as a protein uniquely expressed when MnO2 serves as electron acceptor suggests its utility as a biomarker for MnO2 reduction. This example demonstrates the value of the LC-MS/MS approach for identifying specific proteins of interest and making functional assignments

  2. Comparison of lactate, formate, and propionate as hydrogen donors for the reductive dehalogenation of trichloroethene in a continuous-flow column

    Science.gov (United States)

    Azizian, Mohammad F.; Marshall, Ian P. G.; Behrens, Sebastian; Spormann, Alfred M.; Semprini, Lewis

    2010-04-01

    A continuous-flow column study was conducted to analyze the reductive dehalogenation of trichloroethene (TCE) with aquifer material with high content of iron oxides. The column was bioaugmented with the Point Mugu (PM) culture, which is a mixed microbial enrichment culture capable of completely transforming TCE to ethene (ETH). We determined whether lactate, formate, or propionate fermentation resulted in more effective dehalogenation. Reductive dehalogenation, fermentation, and sulfate, Fe(III), and Mn(IV) reduction were all exhibited within the column. Different steady-states of dehalogenation were achieved based on the concentration of substrates added, with effective transformation to ETH obtained when ample electron donor equivalents were provided. Most of the metabolic reducing equivalents were channeled to sulfate, Fe(III), and Mn(IV) reduction. When similar electron reducing equivalents were added, the most effective dehalogenation was achieved with formate, with 14% of the electron equivalents going towards dehalogenation reactions, compared to 6.5% for lactate and 9.6% for propionate. Effective dehalogenation was maintained over 1000 days of column operation. Over 90% of electron equivalents added could be accounted for by the different electron accepting processes in the column, with 50% associated with soluble and precipitated Fe(II) and Mn(II). Bulk Fe(III) and Mn(IV) reduction was rather associated with lactate and propionate addition than formate addition. Sulfate reduction was a competing electron acceptor reaction with all three electron donors. DNA was extracted from solid coupon samples obtained during the course of the experiment and analyzed using 16S rRNA gene clone libraries and quantitative PCR. Lactate and propionate addition resulted in a significant increase in Geobacter, Spirochaetes, and Desulfitobacterium phylotypes relative to " Dehalococcoides" when compared to formate addition. Results from the molecular biological analyses support

  3. Multi-Scale Monitoring and Prediction of System Responses to Biostimulation

    International Nuclear Information System (INIS)

    Susan Hubbard; Jill Banfield; Jinsong Chen; Mark Conrad; Jenny Druhan; Andreas Englert; Andreas Kemna; LiLi; Phil Long; Michael O'Brien; Dimtrios Ntarlagiannis; Yves Personna; Steve Pride; Lee Slater; Carl Steefel; Ken William

    2007-01-01

    To advance solutions needed for remediation of DOE contaminated sites, approaches are needed that can elucidate and predict reactions associated with coupled biological, geochemical, and hydrological processes over a variety of spatial scales and in heterogeneous environments. Our previous laboratory experimental experiments, which were conducted under controlled and homogeneous conditions, suggest that geophysical methods have the potential for elucidating system transformations that often occur during remediation. Examples include tracking the onset and aggregation of precipitates associated with sulfate reduction using seismic and complex resistivity methods (Williams et al., 2005; Ntarlagiannis et al., 2005) as well as estimating the volume of evolved gas associated with denitrification using radar velocity. These exciting studies illustrated that geophysical responses correlated with biogeochemical changes, but also that multiple factors could impact the geophysical signature and thus a better understanding as well as integration tools were needed to advance the techniques to the point where they can be used to provide quantitative estimates of system transformations. Our current research includes theoretical, numerical, and experimental investigations, performed at the laboratory and the field scales, to determine if geophysical methods can be used to uniquely monitor system transformations. Our work is geared toward the Uranium Mill Tailings site at Rifle, CO, site (Figure 1), where ERSP-sponsored investigations are exploring the efficacy of electron-donor amendments for facilitating sustainable microbial reduction of U(VI) to U(IV) through a series of local-scale field experiments conducted in 2002-2005 (Anderson et al., 2003; Vrionis et al, 2005) Early experiments at the site showed that U(VI) loss from groundwater occurred synchronously with growth of Geobacter after acetate amendment, and illustrated the importance of maintaining iron-reducing conditions

  4. Spatial and temporal dynamics of organohalide-respiring bacteria in a heterogeneous PCE-DNAPL source zone.

    Science.gov (United States)

    Cápiro, Natalie L; Löffler, Frank E; Pennell, Kurt D

    2015-11-01

    Effective treatment of sites contaminated with dense non-aqueous phase liquids (DNAPLs) requires detailed understanding of the microbial community responses to changes in source zone strength and architecture. Changes in the spatial and temporal distributions of the organohalide-respiring Dehalococcoides mccartyi (Dhc) strains and Geobacter lovleyi strain SZ (GeoSZ) were examined in a heterogeneous tetrachloroethene- (PCE-) DNAPL source zone within a two-dimensional laboratory-scale aquifer flow cell. As part of a combined remedy approach, flushing with 2.3 pore volumes (PVs) of 4% (w/w) solution of the nonionic, biodegradable surfactant Tween® 80 removed 55% of the initial contaminant mass, and resulted in a PCE-DNAPL distribution that contained 51% discrete ganglia and 49% pools (ganglia-to-pool ratio of 1.06). Subsequent bioaugmentation with the PCE-to-ethene-dechlorinating consortium BDI-SZ resulted in cis-1,2-dichloroethene (cis-DCE) formation after 1 PV (ca. 7 days), while vinyl chloride (VC) and ethene were detected 10 PVs after bioaugmentation. Maximum ethene yields (ca. 90 μM) within DNAPL pool and ganglia regions coincided with the detection of the vcrA reductive dehalogenase (RDase) gene that exceeded the Dhc 16S rRNA genes by 2.0±1.3 and 4.0±1.7 fold in the pool and ganglia regions, respectively. Dhc and GeoSZ cell abundance increased by up to 4 orders-of-magnitude after 28 PVs of steady-state operation, with 1 to 2 orders-of-magnitude increases observed in close proximity to residual PCE-DNAPL. These observations suggest the involvement of these dechlorinators the in observed PCE dissolution enhancements of up to 2.3 and 6.0-fold within pool and ganglia regions, respectively. Analysis of the solid and aqueous samples at the conclusion of the experiment revealed that the highest VC (≥155 μM) and ethene (≥65 μM) concentrations were measured in zones where Dhc and GeoSZ were predominately attached to the solids. These findings demonstrate

  5. Fermentative Bacteria Influence the Competition between Denitrifiers and DNRA Bacteria

    Directory of Open Access Journals (Sweden)

    Eveline M. van den Berg

    2017-09-01

    Full Text Available Denitrification and dissimilatory reduction to ammonium (DNRA are competing nitrate-reduction processes that entail important biogeochemical consequences for nitrogen retention/removal in natural and man-made ecosystems. The nature of the available carbon source and electron donor have been suggested to play an important role on the outcome of this microbial competition. In this study, the influence of lactate as fermentable carbon source on the competition for nitrate was investigated for varying ratios of lactate and nitrate in the influent (Lac/N ratio. The study was conducted in an open chemostat culture, enriched from activated sludge, under strict anoxia. The mechanistic explanation of the conversions observed was based on integration of results from specific batch tests with biomass from the chemostat, molecular analysis of the biomass enriched, and a computational model. At high Lac/N ratio (2.97 mol/mol both fermentative and respiratory nitrate reduction to ammonium occurred, coupled to partial oxidation of lactate to acetate, and to acetate oxidation respectively. Remaining lactate was fermented to propionate and acetate. At a decreased Lac/N ratio (1.15 mol/mol, the molar percentage of nitrate reduced to ammonium decreased to 58%, even though lactate was supplied in adequate amounts for full ammonification and nitrate remained the growth limiting compound. Data evaluation at this Lac/N ratio suggested conversions were comparable to the higher Lac/N ratio, except for lactate oxidation to acetate that was coupled to denitrification instead of ammonification. Respiratory DNRA on acetate was likely catalyzed by two Geobacter species related to G. luticola and G. lovleyi. Two Clostridiales members were likely responsible for lactate fermentation and partial lactate fermentation to acetate coupled to fermentative DNRA. An organism related to Propionivibrio militaris was identified as the organism likely responsible for denitrification. The

  6. In Situ Bioreduction of Uranium (VI) to Submicromolar Levels and Reoxidation by Dissolved Oxygen

    International Nuclear Information System (INIS)

    Wu, Weimin; Carley, Jack M.; Luo, Jian; Ginder-Vogel, Matthew A.; Cardenas, Erick; Leigh, Mary Beth; Hwang, Chaichi; Kelly, Shelly D.; Ruan, Chuanmin; Wu, Liyou; Van Nostrand, Joy; Gentry, Terry J.; Lowe, Kenneth Alan; Mehlhorn, Tonia L.; Carroll, Sue L.; Luo, Wensui; Fields, Matthew Wayne; Gu, Baohua; Watson, David B.; Kemner, Kenneth M.; Marsh, Terence; Tiedje, James; Zhou, Jizhong; Fendorf, Scott; Kitanidis, Peter K.; Jardine, Philip M.; Criddle, Craig

    2007-01-01

    Groundwater within Area 3 of the U.S. Department of Energy (DOE) Environmental Remediation Sciences Program (ERSP) Field Research Center at Oak Ridge, TN (ORFRC) contains up to 135 (micro)M uranium as U(VI). Through a series of experiments at a pilot scale test facility, we explored the lower limits of groundwater U(VI) that can be achieved by in-situ biostimulation and the effects of dissolved oxygen on immobilized uranium. Weekly 2 day additions of ethanol over a 2-year period stimulated growth of denitrifying, Fe(III)-reducing, and sulfate-reducing bacteria, and immobilization of uranium as U(IV), with dissolved uranium concentrations decreasing to low levels. Following sulfite addition to remove dissolved oxygen, aqueous U(VI) concentrations fell below the U.S. Environmental Protection Agency maximum contaminant limit (MCL) for drinking water ( -1 or 0.126 (micro)M). Under anaerobic conditions, these low concentrations were stable, even in the absence of added ethanol. However, when sulfite additions stopped, and dissolved oxygen (4.0-5.5 mg L -1 ) entered the injection well, spatially variable changes in aqueous U(VI) occurred over a 60 day period, with concentrations increasing rapidly from <0.13 to 2.0 (micro)M at a multilevel sampling (MLS) well located close to the injection well, but changing little at an MLS well located further away. Resumption of ethanol addition restored reduction of Fe(III), sulfate, and U(VI) within 36 h. After 2 years of ethanol addition, X-ray absorption near-edge structure spectroscopy (XANES) analyses indicated that U(IV) comprised 60-80% of the total uranium in sediment samples. At the completion of the project (day 1260), U concentrations in MLS wells were less than 0.1 (micro)M. The microbial community at MLS wells with low U(VI) contained bacteria that are known to reduce uranium, including Desulfovibrio spp. and Geobacter spp., in both sediment and groundwater. The dominant Fe(III)-reducing species were Geothrix spp

  7. Biotransformation involved in sustained reductive removal of uranium in contaminant aquifers

    International Nuclear Information System (INIS)

    Lovley, Derek R.

    2005-01-01

    This report summarizes progress made from August 2004 to July 2005. During this period research focused primarily on obtaining a better understanding of the factors controlling the reduction of U(VI) during in situ uranium bioremediation as well as investigating the potential for using electrodes as an alternative electron donor to promote in situ uranium reduction. Analysis of the 2003 experiment at the field study site in Rifle, CO was completed. The results demonstrated the substantial heterogeneity of the zone undergoing bioremediation, both in terms of geochemistry and microbiology. The lack of U(VI) reduction under sulfate-reducing conditions was clearly documented. The need for more detailed sampling both with time and with depth in the aquifer was demonstrated. For the first time a comparison between the composition of the microbial community in the sediments and the microbes in the corresponding groundwater was attempted. The findings from this study are important not only in further demonstrating the potential for in situ uranium bioremediation, but also for indicating how methods and sampling approaches should be improved in the future. A manuscript summarizing these findings has been accepted for publication in Applied and Environmental Microbiology. In summer of 2004 a new field experiment was conducted at the Rifle site. A novel feature of this study was much more intensive sampling in order to better define the progression of microbial processes during in situ uranium bioremediation. The results demonstrated that stimulation of in situ uranium bioremediation with added acetate was a repeatable phenomenon and that U(VI) reduction was clearly linked to the presence and activity of microorganisms in the family Geobacteraceae. A manuscript summarizing these results is in preparation. A surprising result of the field studies at the Rifle site was that although Geobacter species actively reduced U(VI) in the groundwater, removing it from solution, a high

  8. Comparative Genomic Analysis of Neutrophilic Iron(II Oxidizer Genomes for Candidate Genes in Extracellular Electron Transfer

    Directory of Open Access Journals (Sweden)

    Shaomei He

    2017-08-01

    Full Text Available Extracellular electron transfer (EET is recognized as a key biochemical process in circumneutral pH Fe(II-oxidizing bacteria (FeOB. In this study, we searched for candidate EET genes in 73 neutrophilic FeOB genomes, among which 43 genomes are complete or close-to-complete and the rest have estimated genome completeness ranging from 5 to 91%. These neutrophilic FeOB span members of the microaerophilic, anaerobic phototrophic, and anaerobic nitrate-reducing FeOB groups. We found that many microaerophilic and several anaerobic FeOB possess homologs of Cyc2, an outer membrane cytochrome c originally identified in Acidithiobacillus ferrooxidans. The “porin-cytochrome c complex” (PCC gene clusters homologous to MtoAB/PioAB are present in eight FeOB, accounting for 19% of complete and close-to-complete genomes examined, whereas PCC genes homologous to OmbB-OmaB-OmcB in Geobacter sulfurreducens are absent. Further, we discovered gene clusters that may potentially encode two novel PCC types. First, a cluster (tentatively named “PCC3” encodes a porin, an extracellular and a periplasmic cytochrome c with remarkably large numbers of heme-binding motifs. Second, a cluster (tentatively named “PCC4” encodes a porin and three periplasmic multiheme cytochromes c. A conserved inner membrane protein (IMP encoded in PCC3 and PCC4 gene clusters might be responsible for translocating electrons across the inner membrane. Other bacteria possessing PCC3 and PCC4 are mostly Proteobacteria isolated from environments with a potential niche for Fe(II oxidation. In addition to cytochrome c, multicopper oxidase (MCO genes potentially involved in Fe(II oxidation were also identified. Notably, candidate EET genes were not found in some FeOB, especially the anaerobic ones, probably suggesting EET genes or Fe(II oxidation mechanisms are different from the searched models. Overall, based on current EET models, the search extends our understanding of bacterial EET and

  9. Initial development and structure of biofilms on microbial fuel cell anodes

    Directory of Open Access Journals (Sweden)

    Keller Jürg

    2010-04-01

    Full Text Available Abstract Background Microbial fuel cells (MFCs rely on electrochemically active bacteria to capture the chemical energy contained in organics and convert it to electrical energy. Bacteria develop biofilms on the MFC electrodes, allowing considerable conversion capacity and opportunities for extracellular electron transfer (EET. The present knowledge on EET is centred around two Gram-negative models, i.e. Shewanella and Geobacter species, as it is believed that Gram-positives cannot perform EET by themselves as the Gram-negatives can. To understand how bacteria form biofilms within MFCs and how their development, structure and viability affects electron transfer, we performed pure and co-culture experiments. Results Biofilm viability was maintained highest nearer the anode during closed circuit operation (current flowing, in contrast to when the anode was in open circuit (soluble electron acceptor where viability was highest on top of the biofilm, furthest from the anode. Closed circuit anode Pseudomonas aeruginosa biofilms were considerably thinner compared to the open circuit anode (30 ± 3 μm and 42 ± 3 μm respectively, which is likely due to the higher energetic gain of soluble electron acceptors used. The two Gram-positive bacteria used only provided a fraction of current produced by the Gram-negative organisms. Power output of co-cultures Gram-positive Enterococcus faecium and either Gram-negative organisms, increased by 30-70% relative to the single cultures. Over time the co-culture biofilms segregated, in particular, Pseudomonas aeruginosa creating towers piercing through a thin, uniform layer of Enterococcus faecium. P. aeruginosa and E. faecium together generated a current of 1.8 ± 0.4 mA while alone they produced 0.9 ± 0.01 and 0.2 ± 0.05 mA respectively. Conclusion We postulate that this segregation may be an essential difference in strategy for electron transfer and substrate capture between the Gram-negative and the Gram

  10. Temporal Microbial Community Dynamics in Microbial Electrolysis Cells – Influence of Acetate and Propionate Concentration

    KAUST Repository

    Rao, Hari Ananda

    2017-07-20

    Microbial electrolysis cells (MECs) are widely considered as a next generation wastewater treatment system. However, fundamental insight on the temporal dynamics of microbial communities associated with MEC performance under different organic types with varied loading concentrations is still unknown, nevertheless this knowledge is essential for optimizing this technology for real-scale applications. Here, the temporal dynamics of anodic microbial communities associated with MEC performance was examined at low (0.5 g COD/L) and high (4 g COD/L) concentrations of acetate or propionate, which are important intermediates of fermentation of municipal wastewaters and sludge. The results showed that acetate-fed reactors exhibited higher performance in terms of maximum current density (I: 4.25 ± 0.23 A/m), coulombic efficiency (CE: 95 ± 8%), and substrate degradation rate (98.8 ± 1.2%) than propionate-fed reactors (I: 2.7 ± 0.28 A/m; CE: 68 ± 9.5%; substrate degradation rate: 84 ± 13%) irrespective of the concentrations tested. Despite of the repeated sampling of the anodic biofilm over time, the high-concentration reactors demonstrated lower and stable performance in terms of current density (I: 1.1 ± 0.14 to 4.2 ± 0.21 A/m), coulombic efficiency (CE: 44 ± 4.1 to 103 ± 7.2%) and substrate degradation rate (64.9 ± 6.3 to 99.7 ± 0.5%), while the low-concentration reactors produced higher and dynamic performance (I: 1.1 ± 0.12 to 4.6 ± 0.1 A/m; CE: 52 ± 2.5 to 105 ± 2.7%; substrate degradation rate: 87.2 ± 0.2 to 99.9 ± 0.06%) with the different substrates tested. Correlating reactor\\'s performance with temporal dynamics of microbial communities showed that relatively similar anodic microbial community composition but with varying relative abundances was observed in all the reactors despite differences in the substrate and concentrations tested. Particularly, Geobacter was the predominant bacteria on the anode biofilm of all MECs over time suggesting its

  11. Microbially enhanced dissolution and reductive dechlorination of PCE by a mixed culture: Model validation and sensitivity analysis

    Science.gov (United States)

    Chen, Mingjie; Abriola, Linda M.; Amos, Benjamin K.; Suchomel, Eric J.; Pennell, Kurt D.; Löffler, Frank E.; Christ, John A.

    2013-08-01

    Reductive dechlorination catalyzed by organohalide-respiring bacteria is often considered for remediation of non-aqueous phase liquid (NAPL) source zones due to cost savings, ease of implementation, regulatory acceptance, and sustainability. Despite knowledge of the key dechlorinators, an understanding of the processes and factors that control NAPL dissolution rates and detoxification (i.e., ethene formation) is lacking. A recent column study demonstrated a 5-fold cumulative enhancement in tetrachloroethene (PCE) dissolution and ethene formation (Amos et al., 2009). Spatial and temporal monitoring of key geochemical and microbial (i.e., Geobacter lovleyi and Dehalococcoides mccartyi strains) parameters in the column generated a data set used herein as the basis for refinement and testing of a multiphase, compositional transport model. The refined model is capable of simulating the reactive transport of multiple chemical constituents produced and consumed by organohalide-respiring bacteria and accounts for substrate limitations and competitive inhibition. Parameter estimation techniques were used to optimize the values of sensitive microbial kinetic parameters, including maximum utilization rates, biomass yield coefficients, and endogenous decay rates. Comparison and calibration of model simulations with the experimental data demonstrate that the model is able to accurately reproduce measured effluent concentrations, while delineating trends in dechlorinator growth and reductive dechlorination kinetics along the column. Sensitivity analyses performed on the optimized model parameters indicate that the rates of PCE and cis-1,2-dichloroethene (cis-DCE) transformation and Dehalococcoides growth govern bioenhanced dissolution, as long as electron donor (i.e., hydrogen flux) is not limiting. Dissolution enhancements were shown to be independent of cis-DCE accumulation; however, accumulation of cis-DCE, as well as column length and flow rate (i.e., column residence time

  12. Microbial Community Response to Carbon Substrate Amendment in Mercury Impacted Sediments: Implications on Microbial Methylation of Mercury.

    Science.gov (United States)

    Elias, D. A.; Somenahally, A. C.; Moberly, J. G.; Hurt, R. A., Jr.; Brown, S. D.; Podar, M.; Palumbo, A. V.; Gilmour, C. C.

    2015-12-01

    Methylmercury (MeHg) is a neurotoxic and bio-accumulative product of the microbial methylation of inorganic mercury (Hg(II)). Methylating organisms are now known to exist in almost all anaerobic niches including fermentation, Fe(III)- and sulfate- reduction as well as methanogenesis. The study objective was to determine the effect of different carbon sources on the microbial community and methylating populations in particular along a Hg contaminated creek. Sediment cores from upstream and downstream at the Hg contaminated East Fork Poplar Creek (EFPC), Oak Ridge TN, and a background site were sectioned by depth, and Hg-methylation potential (HgMP) assays were performed using stable isotope spikes. Sediments from the lowest depth possessed the highest in-situ activity. Replicate samples were amended with different carbon substrates (cellulose, acetate, propionate, lactate, ethanol and methanol), spiked with stable isotopes for HgMP assays and incubated for 24hrs. Sequencing of the 16S rRNA gene was performed to determine alterations in Bacterial and Archaeal population dynamics. Additionally, bioinformatics and our new qualitative and quantitative hgcAB primers were utilized to determine microbial community structure alterations and correlate organism and gene abundance with altered MeHg generation. HgMP was significantly reduced in cellulose amended sediments while acetate and propionate slightly decreased HgMP in both sites. Methanol, ethanol and lactate increased the HgMP in EFPC downstream while cellulose amendment significantly decreased the Proteobacteria, and the Firmicutes increased but none are currently known to produce MeHg. Geobacter bemidjiensis in particular significantly decreased in cellulose amended sediments in all three sites from being predominant in-situ. This suggests that in EFPC downstream and background sites, the prevalent Hg-methyaltors might be Deltaprotebacteria, since upstream, cellulose amendment did not reduce HgMP even though

  13. Microbial Populations of Stony Meteorites: Substrate Controls on First Colonizers

    Directory of Open Access Journals (Sweden)

    Alastair W. Tait

    2017-06-01

    Full Text Available Finding fresh, sterilized rocks provides ecologists with a clean slate to test ideas about first colonization and the evolution of soils de novo. Lava has been used previously in first colonizer studies due to the sterilizing heat required for its formation. However, fresh lava typically falls upon older volcanic successions of similar chemistry and modal mineral abundance. Given enough time, this results in the development of similar microbial communities in the newly erupted lava due to a lack of contrast between the new and old substrates. Meteorites, which are sterile when they fall to Earth, provide such contrast because their reduced and mafic chemistry commonly differs to the surfaces on which they land; thus allowing investigation of how community membership and structure respond to this new substrate over time. We conducted 16S rRNA gene analysis on meteorites and soil from the Nullarbor Plain, Australia. We found that the meteorites have low species richness and evenness compared to soil sampled from directly beneath each meteorite. Despite the meteorites being found kilometers apart, the community structure of each meteorite bore more similarity to those of other meteorites (of similar composition than to the community structure of the soil on which it resided. Meteorites were dominated by sequences that affiliated with the Actinobacteria with the major Operational Taxonomic Unit (OTU classified as Rubrobacter radiotolerans. Proteobacteria and Bacteroidetes were the next most abundant phyla. The soils were also dominated by Actinobacteria but to a lesser extent than the meteorites. We also found OTUs affiliated with iron/sulfur cycling organisms Geobacter spp. and Desulfovibrio spp. This is an important finding as meteorites contain abundant metal and sulfur for use as energy sources. These ecological findings demonstrate that the structure of the microbial community in these meteorites is controlled by the substrate, and will not

  14. The microbial ferrous wheel in a neutral pH groundwater seep

    Directory of Open Access Journals (Sweden)

    Eric eRoden

    2012-05-01

    Full Text Available Evidence for microbial Fe redox cycling was documented in a circumneutral pH groundwater seep near Bloomington, Indiana. Geochemical and microbiological analyses were conducted at two sites, a semi-consolidated microbial mat and a floating puffball structure. In situ voltammetric microelectrode measurements revealed steep opposing gradients of O2 and Fe(II at both sites, similar to other groundwater seep and sedimentary environments known to support microbial Fe redox cycling. The puffball structure showed an abrupt increase in dissolved Fe(II just at its surface (~ 5 cm depth, suggesting an internal Fe(II source coupled to active Fe(III reduction. MPN enumerations detected microaerophilic Fe(II-oxidizing bacteria (FeOB and dissimilatory Fe(III-reducing bacteria (FeRB at densities of 102-105 cells mL-1 in samples from both sites. In vitro Fe(III reduction experiments revealed the potential for immediate reduction (no lag period of native Fe(III oxides. Conventional full-length 16S rRNA gene clone libraries were compared withhigh throughput barcode sequencing of the V1, V4 or V6 variable regions of 16S rRNA genes in order to evaluate the extent to which new sequencing approaches could provide enhanced insight into the composition of Fe redox cycling microbial community structure. The composition of the clone libraries suggested a lithotroph-dominated microbial community centered around taxa related to known FeOB (e.g. Gallionella, Sideroxydans, Aquabacterium. Sequences related to recognized FeRB (e.g. Rhodoferax, Aeromonas, Geobacter, Desulfovibrio were also well represented. Overall, sequences related to known FeOB and FeRB accounted for 88 and 59% of total clone sequences in the mat and puffball libraries, respectively. Taxa identified in the barcode libraries showed partial overlap with the clone libraries, but were not always consistent across different variable regions and sequencing platforms. However, the barcode libraries provided

  15. Role of Sulfhydryl Sites on Bacterial Cell Walls in the Biosorption, Mobility and Bioavailability of Mercury and Uranium

    Energy Technology Data Exchange (ETDEWEB)

    Myneni, Satish C. [Princeton Univ., NJ (United States); Mishra, Bhoopesh [Princeton Univ., NJ (United States); Fein, Jeremy [Princeton Univ., NJ (United States)

    2009-04-01

    The goal of this exploratory study is to provide a quantitative and mechanistic understanding of the impact of bacterial sulfhydryl groups on the bacterial uptake, speciation, methylation and bioavailability of Hg and redox changes of uranium. The relative concentration and reactivity of different functional groups present on bacterial surfaces will be determined, enabling quantitative predictions of the role of biosorption of Hg under the physicochemical conditions found at contaminated DOE sites.The hypotheses we propose to test in this investigation are as follows- 1) Sulfhydryl groups on bacterial cell surfaces modify Hg speciation and solubility, and play an important role, specifically in the sub-micromolar concentration ranges of metals in the natural and contaminated systems. 2) Sulfhydryl binding of Hg on bacterial surfaces significantly influences Hg transport into the cell and the methylation rates by the bacteria. 3) Sulfhydryls on cell membranes can interact with hexavalent uranium and convert to insoluble tetravalent species. 4) Bacterial sulfhydryl surface groups are inducible by the presence of metals during cell growth. Our studies focused on the first hypothesis, and we examined the nature of sulfhydryl sites on three representative bacterial species: Bacillus subtilis, a common gram-positive aerobic soil species; Shewanella oneidensis, a facultative gram-negative surface water species; and Geobacter sulfurreducens, an anaerobic iron-reducing gram-negative species that is capable of Hg methylation; and at a range of Hg concentration (and Hg:bacterial concentration ratio) in which these sites become important. A summary of our findings is as follows- Hg adsorbs more extensively to bacteria than other metals. Hg adsorption also varies strongly with pH and chloride concentration, with maximum adsorption occurring under circumneutral pH conditions for both Cl-bearing and Cl-free systems. Under these conditions, all bacterial species tested exhibit

  16. Harnessing microbial subsurface metal reduction activities to synthesise nanoscale cobalt ferrite with enhanced magnetic properties

    Energy Technology Data Exchange (ETDEWEB)

    Coker, Victoria S.; Telling, Neil D.; van der Laan, Gerrit; Pattrick, Richard A.D.; Pearce, Carolyn I.; Arenholz, Elke; Tuna, Floriana; Winpenny, Richard E.P.; Lloyd, Jonathan R.

    2009-03-24

    Nanoscale ferrimagnetic particles have a diverse range of uses from directed cancer therapy and drug delivery systems to magnetic recording media and transducers. Such applications require the production of monodisperse nanoparticles with well-controlled size, composition, and magnetic properties. To fabricate these materials purely using synthetic methods is costly in both environmental and economical terms. However, metal-reducing microorganisms offer an untapped resource to produce these materials. Here, the Fe(III)-reducing bacterium Geobacter sulfurreducens is used to synthesize magnetic iron oxide nanoparticles. A combination of electron microscopy, soft X-ray spectroscopy, and magnetometry techniques was employed to show that this method of biosynthesis results in high yields of crystalline nanoparticles with a narrow size distribution and magnetic properties equal to the best chemically synthesized materials. In particular, it is demonstrated here that cobalt ferrite (CoFe{sub 2}O{sub 4}) nanoparticles with low temperature coercivity approaching 8 kOe and an effective anisotropy constant of {approx} 10{sup 6} erg cm{sup -3} can be manufactured through this biotechnological route. The dramatic enhancement in the magnetic properties of the nanoparticles by the introduction of high quantities of Co into the spinel structure represents a significant advance over previous biomineralization studies in this area using magnetotactic bacteria. The successful production of nanoparticulate ferrites achieved in this study at high yields could open up the way for the scaled-up industrial manufacture of nanoparticles using environmentally benign methodologies. Production of ferromagnetic nanoparticles for pioneering cancer therapy, drug delivery, chemical sensors, catalytic activity, photoconductive materials, as well as more traditional uses in data storage embodies a large area of inorganic synthesis research. In particular, the addition of transition metals other than

  17. Harnessing microbial subsurface metal reduction activities to synthesize nanoscale cobalt ferrite with enhanced magnetic properties

    International Nuclear Information System (INIS)

    Coker, Victoria S.; Telling, Neil D.; van der Laan, Gerrit; Pattrick, Richard A.D.; Pearce, Carolyn I.; Arenholz, Elke; Tuna, Floriana; Winpenny, Richard E.P.; Lloyd, Jonathan R.

    2009-01-01

    Nanoscale ferrimagnetic particles have a diverse range of uses from directed cancer therapy and drug delivery systems to magnetic recording media and transducers. Such applications require the production of monodisperse nanoparticles with well-controlled size, composition, and magnetic properties. To fabricate these materials purely using synthetic methods is costly in both environmental and economical terms. However, metal-reducing microorganisms offer an untapped resource to produce these materials. Here, the Fe(III)-reducing bacterium Geobacter sulfurreducens is used to synthesize magnetic iron oxide nanoparticles. A combination of electron microscopy, soft X-ray spectroscopy, and magnetometry techniques was employed to show that this method of biosynthesis results in high yields of crystalline nanoparticles with a narrow size distribution and magnetic properties equal to the best chemically synthesized materials. In particular, it is demonstrated here that cobalt ferrite (CoFe 2 O 4 ) nanoparticles with low temperature coercivity approaching 8 kOe and an effective anisotropy constant of ∼ 10 6 erg cm -3 can be manufactured through this biotechnological route. The dramatic enhancement in the magnetic properties of the nanoparticles by the introduction of high quantities of Co into the spinel structure represents a significant advance over previous biomineralization studies in this area using magnetotactic bacteria. The successful production of nanoparticulate ferrites achieved in this study at high yields could open up the way for the scaled-up industrial manufacture of nanoparticles using environmentally benign methodologies. Production of ferromagnetic nanoparticles for pioneering cancer therapy, drug delivery, chemical sensors, catalytic activity, photoconductive materials, as well as more traditional uses in data storage embodies a large area of inorganic synthesis research. In particular, the addition of transition metals other than Fe into the structure

  18. Sustainable resource recovery and energy conversion processes using microbial electrochemical technologies

    Science.gov (United States)

    Yates, Matthew D.

    Microbial Electrochemical Technologies (METs) are emerging technological platforms for the conversion of waste into usable products. METs utilize naturally occurring bacteria, called exoelectrogens, capable of transferring electrons to insoluble terminal electron acceptors. Electron transfer processes in the exoelectrogen Geobacter sulfurreducens were exploited here to develop sustainable processes for synthesis of industrially and socially relevant end products. The first process examined was the removal of soluble metals from solution to form catalytic nanoparticles and nanoporous structures. The second process examined was the biocatalytic conversion of electrons into hydrogen gas using electrons supplied directly to an electrode. Nanoparticle formation is desirable because materials on the nanoscale possess different physical, optical, electronic, and mechanical properties compared to bulk materials. In the first process, soluble palladium was used to form catalytic palladium nanoparticles using extracellular electron transfer (EET) processes of G. sulfurreducens, typically the dominant member of mixedculture METs. Geobacter cells reduced the palladium extracellularly using naturally produced pili, which provided extracellular adsorption and reduction sites to help delay the diffusion of soluble metals into the cell. The extracellular reduction prevented cell inactivation due to formation of intracellular particles, and therefore the cells could be reused in multiple palladium reduction cycles. A G. sulfurreducens biofilm was next investigated as a biotemplate for the formation of a nanoporous catalytic palladium structure. G. sulfurreducens biofilms have a dense network of pili and extracellular cytochromes capable of high rates of electron transfer directly to an electrode surface. These pili and cytochromes provide a dense number of reduction sites for nanoparticle formation without the need for any synthetic components. The cells within the biofilm also can

  19. An Updated Model for the Anomalous Resistivity of LNAPL Plumes in Sandy Environments

    Science.gov (United States)

    Sauck, W. A.; Atekwana, E. A.; Werkema, D. D.

    2006-05-01

    and the soil samples for bacterial assays. Those columns to which diesel oil had been added (4) clearly showed the resistivity to decrease, the TDS of water samples to increase, and a radical change in bacterial populations favoring the oil degraders to occur. In the laboratory, most of the significant changes occurred in the time span of 4 to 6 months. The third development that may have a bearing on anomalous conductivity was the recent discovery that certain bacteria, (e.g., Geobacter sulfurreducens) have hairlike structures called pili that are excellent electrical conductors. These pili (nano-wires), longer than the bacterial body itself, are apparently used for electron transfer by the bacteria. In sufficient numbers, they could conceivably alter the electrical resistivity of a volume of earth. In summary, our understanding for the cause of anomalous conductive zones associated with areas impacted by LNAPL spills has been significantly improved by these recent developments.

  20. Characterization of microbial communities in former neutral uranium mines in Saxony and studies on the microbial immobilization of uranium and arsenic

    International Nuclear Information System (INIS)

    Gagell, Corinna

    2015-01-01

    Abandoned uranium mines contribute significantly to the emission of contaminants such as uranium and arsenic into partly densely populated regions due to emerging flood water. To get a deeper understanding of ongoing processes in underground environments and for the development of alternative strategies to conventional, cost-intensive water treatment, the objective of this thesis was to characterize microbial communities from three former uranium mines in Saxony, namely Poehla, Schlema, and Zobes representing different flooding stages and to investigate the microbial influence on the mobility of uranium and arsenic. To find out which microorganisms could affect hydrochemical processes in underground environments, the diversity and compostion of microbial communities was investigated by pyrosequencing of a 16S rRNA gene (16S rDNA) fragment together with CARD-FISH. Though cluster analyses showed that planktonic communities differed with regard to bacterial composition between the three uranium mines, all were dominated by chemolithotrophic sulfur oxidizers of Betaproteobacteria with members of genus Thiobacillus and Sulfuritalea and Epsilonproteobacteria belonging to Sulfuricurvum and Sulfurimonas. Unlike planktonic communities, in situ biofilms grown on BACTRAPs during three month of exposition in flood water consisted of metal and sulfate reducing Deltaproteobacteria to a substantial or even dominant proportion based on pyrosequencing results. In biofilm communities from Zobes mainly Geobacter sp. were detected which are known Fe(III)- and U(VI)-reducing bacteria. Although CARD-FISH analysis revealed that Archaea represented only a very small part of the planktonic communities, planktonic Euryarchaeota of the Thermoprotei class were detected in all mines by pyrosequencing. In planktonic communities and 3-month biofilms of Poehla and Zobes methanogenic Crenarchaeota, especially Methanobacteria and partially Methanomicrobia, were determined, too. 16S rRNA analysis

  1. Diversity and ubiquity of bacteria capable of utilizing humic substances as electron donors for anaerobic respiration.

    Science.gov (United States)

    Coates, John D; Cole, Kimberly A; Chakraborty, Romy; O'Connor, Susan M; Achenbach, Laurie A

    2002-05-01

    Previous studies have demonstrated that reduced humic substances (HS) can be reoxidized by anaerobic bacteria such as Geobacter, Geothrix, and Wolinella species with a suitable electron acceptor; however, little is known of the importance of this metabolism in the environment. Recently we investigated this metabolism in a diversity of environments including marine and aquatic sediments, forest soils, and drainage ditch soils. Most-probable-number enumeration studies were performed using 2,6-anthrahydroquinone disulfonate (AHDS), an analog for reduced HS, as the electron donor with nitrate as the electron acceptor. Anaerobic organisms capable of utilizing reduced HS as an electron donor were found in all environments tested and ranged from a low of 2.31 x 10(1) in aquifer sediments to a high of 9.33 x 10(6) in lake sediments. As part of this study we isolated six novel organisms capable of anaerobic AHDS oxidation. All of the isolates coupled the oxidation of AHDS to the reduction of nitrate with acetate (0.1 mM) as the carbon source. In the absence of cells, no AHDS oxidation was apparent, and in the absence of AHDS, no cell density increase was observed. Generally, nitrate was reduced to N(2). Analysis of the AHDS and its oxidized form, 2,6-anthraquinone disulfonate (AQDS), in the medium during growth revealed that the anthraquinone was not being biodegraded as a carbon source and was simply being oxidized as an energy source. Determination of the AHDS oxidized and nitrate reduced accounted for 109% of the theoretical electron transfer. In addition to AHDS, all of these isolates could also couple the oxidation of reduced humic substances to the reduction of nitrate. No HS oxidation occurred in the absence of cells and in the absence of a suitable electron acceptor, demonstrating that these organisms were capable of utilizing natural HS as an energy source and that AHDS serves as a suitable analog for studying this metabolism. Alternative electron donors included

  2. Bacterial Community Analysis, New Exoelectrogen Isolation and Enhanced Performance of Microbial Electrochemical Systems Using Nano-Decorated Anodes

    Science.gov (United States)

    Xu, Shoutao

    Microbial electrochemical systems (MESs) have attracted much research attention in recent years due to their promising applications in renewable energy generation, bioremediation, and wastewater treatment. In a MES, microorganisms interact with electrodes via electrons, catalyzing oxidation and reduction reactions at the anode and the cathode. The bacterial community of a high power mixed consortium MESs (maximum power density is 6.5W/m2) was analyzed by using denature gradient gel electrophoresis (DGGE) and 16S DNA clone library methods. The bacterial DGGE profiles were relatively complex (more than 10 bands) but only three brightly dominant bands in DGGE results. These results indicated there are three dominant bacterial species in mixed consortium MFCs. The 16S DNA clone library method results revealed that the predominant bacterial species in mixed culture is Geobacter sp (66%), Arcobacter sp and Citrobacter sp. These three bacterial species reached to 88% of total bacterial species. This result is consistent with the DGGE result which showed that three bright bands represented three dominant bacterial species. Exoelectrogenic bacterial strain SX-1 was isolated from a mediator-less microbial fuel cell by conventional plating techniques with ferric citrate as electron acceptor under anaerobic conditions. Phylogenetic analysis of the 16S rDNA sequence revealed that it was related to the members of Citrobacter genus with Citrobacter sp. sdy-48 being the most closely related species. The bacterial strain SX-1 produced electricity from citrate, acetate, glucose, sucrose, glycerol, and lactose in MFCs with the highest current density of 205 mA/m2 generated from citrate. Cyclic voltammetry analysis indicated that membrane associated proteins may play an important role in facilitating electron transfer from the bacteria to the electrode. This is the first study that demonstrates that Citrobacter species can transfer electrons to extracellular electron acceptors

  3. Metaproteomics Identifies the Protein Machinery Involved in Metal and Radionuclide Reduction in Subsurface Microbiomes and Elucidates Mechanisms and U(VI) Reduction Immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Pfiffner, Susan M. [Univ. of Tennessee, Knoxville, TN (United States); Löffler, Frank [Univ. of Tennessee, Knoxville, TN (United States); Ritalahti, Kirsti [Univ. of Tennessee, Knoxville, TN (United States); Sayler, Gary [Univ. of Tennessee, Knoxville, TN (United States); Layton, Alice [Univ. of Tennessee, Knoxville, TN (United States); Hettich, Robert [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2015-08-31

    analyses, and gene expression studies to support the metaproteomics characterizations. Growth experiments of target microorganisms (Anaeromyxobacter, Shewanella, Geobacter) revealed tremendous respiratory versatility, as evidenced by the ability to utilize a range of electron donors (e.g. acetate, hydrogen, pyruvate, lactate, succinate, formate) and electron acceptors (e.g. nitrate, fumarate, halogenated phenols, ferric iron, nitrous oxide, etc.). In particular, the dissimilatory metabolic reduction of metals, including radionuclides, by target microorganisms spurred interest for in situ bioremediation of contaminated soils and sediments. Distinct c-type cytochrome expression patterns were observed in target microorganisms grown with the different electron acceptors. For each target microorganism, the core proteome covered almost all metabolic pathways represented by their corresponding pan-proteomes. Unique proteins were detected for each target microorganism, and their expression and possible functionalities were linked to specific growth conditions through proteomics measurements. Optimization of the proteomic tools included in-depth comprehensive metagenomic and metaproteomic analyses on a limited number of samples. The optimized metaproteomic analyses were then applied to Oak Ridge IFRC field samples from the slow-release substrate biostimulation. Metaproteomic analysis and pathway mapping results demonstrated the distinct effects of metal and non-metal growth conditions on the proteome expression. With these metaproteomic tools, we identified which previously hypothetical metabolic pathways were active during the analyzed time points of the slow release substrate biostimulation. Thus, we demonstrated the utility of these tools for site assessment, efficient implementation of bioremediation and long-term monitoring. This research of detailed protein analysis linked with metal reduction activity did (1) show that c-type cytochrome isoforms, previously associated with

  4. Microbiology and optimization of hydrogen fermentation and bioelectricity production

    Energy Technology Data Exchange (ETDEWEB)

    Makinen, A.

    2013-11-01

    yield. Pentose fermentation was accompanied by production of acetate, butyrate and formate, while in hexose fermentation the main soluble end product was lactate. In CSTR Hisarkoy enrichment culture produced H{sub 2} from xylose with the maximum average H{sub 2} yield and production rate of 1.97 mol/mol xylose and 7.3 mmol/h/L, respectively, at suboptimal temperature of 45 deg C for meso- and thermophiles. At 45 deg C microbial community consisted of only two bacterial strains affiliated to Clostridium acetobutylicum and Cirtobacter freundii. An exoeletrogenic culture was enriched on xylose from compost sample in MFCs. In enrichment phase electricity production in MFCs was accompanied with ethanol production. The main bacterium responsible of xylose degradation was xylanolytic Ruminobacillus xylanolyticum and the main bacteria responsible for electricity production were denitrifiers Paracoccus pantotrophus, Comamonas denitrificans and Alicycliphilus denitrificans. Anode potential had a significant effect on current production in MFCs. Compost enrichment culture was able to produce electricity from xylose at poised anode potential of 0.4 V vs standard hydrogen electrode (SHE) having the maximum current density and Coulombic efficiency (CE) of 1.65 A/m{sup 2} and 37 %, respectively. Lower anode potentials of 0.1 or -0.2 V vs SHE didn't lead to current production. Optimum operational parameters for bioelectricity production from xylose by compost enrichment culture were without mixing, external resistance of 100 ohm, 0.5 g/L xylose and pH 7. High current density and CE of 1.74 A/m{sup 2} and 82 %, respectively, were obtained. This is the highest CE obtained with xylose in two-chamber MFC reported in the literature. This very efficient exoelectrogenic culture was dominated by Geobacter species, including G. sulfurreducens, which were enriched on anode biofilm. Xylose fermenters, including Escherichia coli, Sphaerochaeta sp. TQ1, and Bacteroides sp. were present in

  5. Hydrogeologic characterization and assessment of bioremediation of chlorinated benzenes and benzene in wetland areas, Standard Chlorine of Delaware, Inc. Superfund Site, New Castle County, Delaware, 2009-12

    Science.gov (United States)

    Lorah, Michelle M.; Walker, Charles W.; Baker, Anna C.; Teunis, Jessica A.; Emily Majcher,; Brayton, Michael J.; Raffensperger, Jeff P.; Cozzarelli, Isabelle M.

    2015-01-01

    desorption from the sediments.When highly reducing, methanogenic, or sulfate-reducing conditions existed in the wetland groundwater, molar composition of the volatile organic compounds (VOCs) showed that chlorobenzene and benzene were predominant, indicating biodegradation of the chlorinated benzenes through reductive dechlorination pathways. Temporal changes in redox conditions between 2009 and 2011–12 have shifted the locations in the wetland study area where reductive dechlorination is evident. Microbial community analyses of sediment showed relatively high cell numbers and diversity of populations (Dehalococcoides, Dehalobacter, Desulfitobacterium, and Geobacter) that are known to contain species capable of reductive dechlorination, confirming groundwater geochemistry evidence of the occurrence of reductive dechlorination. Natural attenuation was not sufficient, however, to reduce total VOC concentrations along upward groundwater flowpaths in the wetland sediments, most likely due to the additional source of contaminants in the upper sediments. In situ microcosms that were unamended except for the addition of 13C-labeled contaminants in some treatments, confirmed that the native microbial community was able to biodegrade the higher chlorinated benzenes through reductive dechlorination and that 1,2-dichlorobenzene, chlorobenzene, and benzene could be degraded to carbon dioxide through oxidation pathways. Microcosms that were bioaugmented with the anaerobic dechlorinating consortium WBC-2 and deployed in the wetland sediments showed reductive dechlorination of tri-, di-, and monochlorobenzene, and 13C-chlorobenzene treatments showed complete degradation of chlorobenzene to carbon dioxide under anaerobic conditions.Experiments with a continuous flow, fixed-film bioreactor seeded with native microorganisms in groundwater from the wetland area showed both aerobic and anaerobic biodegradation of dichlorobenzenes, monochlorobenzene, and benzene, although

  6. Microbial Oxidation of Hg(0) - Its Effect on Hg Stable Isotope Fractionation and Methylmercury Production

    Energy Technology Data Exchange (ETDEWEB)

    Yee, Nathan [Rutgers Univ., New Brunswick, NJ (United States); Barkay, Tamar [Rutgers Univ., New Brunswick, NJ (United States); Reinfelder, John [Rutgers Univ., New Brunswick, NJ (United States)

    2016-06-28

    relationship between Hg concentrations and rates of denitrification in enrichment cultures. In part III of our project, we examined in more detail the effects of microbial interactions on Hg transformations. We discovered that both sulfate reducing and iron reducing bacteria coexist in freshwater sediments and both microbial groups contribute to mercury methylation. We showed that mercury methylation by sulfate reducing and iron reducing bacteria are temporally and spatially separated processes. We also discovered that methanogens can methylate mercury. We showed that Methanospirillum hungatei JF-1 methylated Hg at comparable rates, but with higher yields, than those observed for sulfate-reducing bacteria and iron-reducing bacteria. Finally, we demonstrated that syntrophic interactions between different microbial groups increase mercury methylation rates. We showed that Hg methylation rates are stimulated via inter-species hydrogen and acetate transfer (i) from sulfate-reducing bacteria to methanogens and (ii) from fermenters to the sulfate-reducing bacteria. In part IV of the project, we studied Hg bioavailability and Hg isotope fractionation. We demonstrated that thiol-bound Hg is bioavailable to mercury resistant bacteria. We found that uptake of Hg from Hg-glutathione and Hg-cysteine complexes does not require functioning glutathione and cystine/cysteine transport systems. We demonstrated that a wide range of methylmercury complexes (e.g. MeHgOH, MeHg-cysteine, and MeHg-glutathione) are bioavailable to mercury resistant bacteria. The rate of MeHg demethylation varies more between different species of mercury resistant bacteria than among MeHg complexes. We showed that microbial demethylation of MeHg depends more on the species of microorganism than on the types and relative concentrations of thiols or other MeHg ligands present. Finally, we demonstrated that Hg methylation by Geobacter sulfurreducens PCA and Desulfovibrio desulfuricans ND132 imparts mass