Microbial Nanowires: Is the Subsurface Hardwired ?

The Earth\u27s shallow subsurface results from integrated biological, geochemical, and physical processes. Methods are sought to remotely assess these interactive processes, especially those catalysed by micro-organisms. Using saturated sand columns and the metal reducing bacterium Shewanella oneidensis MR-1, we show that electrically conductive appendages called bacterial nanowires are directly associated with electrical potentials. No significant electrical potentials were detectable in columns inoculated with mutant strains that produced non-conductive appendages. Scanning electron microscopy imaging revealed a network of nanowires linking cells-cells and cells to mineral surfaces, hardwiring the entire length of the column. We hypothesize that the nanowires serve as conduits for transfer of electrons from bacteria in the anaerobic part of the column to bacteria at the surface that have access to oxygen, akin to a biogeobattery. These results advance understanding of the mechanisms of electron transport in subsurface environments and of how microorganisms cycle geologic material and share energy

Composition, Reactivity, and Regulation of Extracellular Metal-Reducing Structures(Nanowires) Produced by Dissimilatory Metal Reducing Bacteria

Image collection of composition, reactivity, and regulation of extracellular metal-reducing structures (nanowires) produced by dissimilatory metal reducing bacteria

Functionally Stable and Phylogenetically Diverse Microbial Enrichments from Microbial Fuel Cells during Wastewater Treatment

Microbial fuel cells (MFCs) are devices that exploit microorganisms as biocatalysts to recover energy from organic matter in the form of electricity. One of the goals of MFC research is to develop the technology for cost-effective wastewater treatment. However, before practical MFC applications are implemented it is important to gain fundamental knowledge about long-term system performance, reproducibility, and the formation and maintenance of functionally-stable microbial communities. Here we report findings from a MFC operated for over 300 days using only primary clarifier effluent collected from a municipal wastewater treatment plant as the microbial resource and substrate. The system was operated in a repeat-batch mode, where the reactor solution was replaced once every two weeks with new primary effluent that consisted of different microbial and chemical compositions with every batch exchange. The turbidity of the primary clarifier effluent solution notably decreased, and 97% of biological oxygen demand (BOD) was removed after an 8–13 day residence time for each batch cycle. On average, the limiting current density was 1000 mA/m2, the maximum power density was 13 mW/m2, and coulombic efficiency was 25%. Interestingly, the electrochemical performance and BOD removal rates were very reproducible throughout MFC operation regardless of the sample variability associated with each wastewater exchange. While MFC performance was very reproducible, the phylogenetic analyses of anode-associated electricity-generating biofilms showed that the microbial populations temporally fluctuated and maintained a high biodiversity throughout the year-long experiment. These results suggest that MFC communities are both self-selecting and self-optimizing, thereby able to develop and maintain functional stability regardless of fluctuations in carbon source(s) and regular introduction of microbial competitors. These results contribute significantly toward the practical application of MFC systems for long-term wastewater treatment as well as demonstrating MFC technology as a useful device to enrich for functionally stable microbial populations

The influence of cultivation methods on Shewanella oneidensis physiology and proteome expression

High-throughput analyses that are central to microbial systems biology and ecophysiology research benefit from highly homogeneous and physiologically well-defined cell cultures. While attention has focused on the technical variation associated with high-throughput technologies, biological variation introduced as a function of cell cultivation methods has been largely overlooked. This study evaluated the impact of cultivation methods, controlled batch or continuous culture in bioreactors versus shake flasks, on the reproducibility of global proteome measurements in Shewanellaoneidensis MR-1. Variability in dissolved oxygen concentration and consumption rate, metabolite profiles, and proteome was greater in shake flask than controlled batch or chemostat cultures. Proteins indicative of suboxic and anaerobic growth (e.g., fumarate reductase and decaheme c-type cytochromes) were more abundant in cells from shake flasks compared to bioreactor cultures, a finding consistent with data demonstrating that “aerobic” flask cultures were O2 deficient due to poor mass transfer kinetics. The work described herein establishes the necessity of controlled cultivation for ensuring highly reproducible and homogenous microbial cultures. By decreasing cell to cell variability, higher quality samples will allow for the interpretive accuracy necessary for drawing conclusions relevant to microbial systems biology research

Electron Transport in the Dissimilatory Iron Reducer, GS-15

Mechanisms for electron transport to Fe(III) were investigated in GS-15, a novel anaerobic microorganism which can obtain energy for growth by coupling the complete oxidation of organic acids or aromatic compounds to the reduction of Fe(III) to Fe(II). The results indicate that Fe(III) reduction proceeds through a type b cytochrome and a membrane-bound Fe(III) reductase which is distinct from the nitrate reductase

Reduction Kinetics of Fe(III), Co(III), U(VI), Cr(VI), and Tc(VII) in Cultures of Dissimilatory Metal-Reducing Bacteria

The reduction kinetics of Fe(III)citrate, Fe(III)NTA, Co(III)EDTA−, U(VI)O22+, Cr(VI)O42−, and Tc(VII)O4− were studied in cultures of dissimilatory metal reducing bacteria (DMRB): Shewanella alga strain BrY, Shewanella putrefaciens strain CN32, Shewanella oneidensis strain MR-1, and Geobacter metallireducens strain GS-15. Reduction rates were metal specific with the following rate trend: Fe (III)citrate ≥ Fe(III)NTA \u3e Co(III)EDTA− ›› UO2 2+ \u3e CrO42− \u3e TcO4−, except for CrO42− when H2 was used as electron donor. The metal reduction rates were also electron donor dependent with faster rates observed for H2 than lactate− for all Shewanella species despite higher initial lactate (10 mM) than H2 (0.48 mM). The bioreduction of CrO42− was anomalously slower compared to the other metals with H2 as an electron donor relative to lactate and reduction ceased before all the CrO42− had been reduced. Transmission electron microscopic (TEM) and energy-dispersive spectroscopic (EDS) analyses performed on selected solids at experiment termination found precipitates of reduced U and Tc in association with the outer cell membrane and in the periplasm of the bacteria. The kinetic rates of metal reduction were correlated with the precipitation of reduced metal phases and their causal relationship discussed. The experimental rate data were well described by a Monod kinetic expression with respect to the electron acceptor for all metals except CrO42−, for which the Monod model had to be modified to account for incomplete reduction. However, the Monod models became statistically over-parametrized, resulting in large uncertainties of their parameters. A first-order approximation to the Monod model also effectively described the experimental results, but the rate coefficients exhibited far less uncertainty. The more precise rate coefficients of the first-order model provided a better means than the Monod parameters, to quantitatively compare the reduction rates between metals, electron donors, and DMRB species

MICROBIALLY PROMOTED SOLUBILIZATION OF STEEL CORROSION PRODUCTS AND FATE OF ASSOCIATED ACTINIDES

The research is designed to develop a safe and effective biological approach for decontaminating mild and stainless steels that were used in the production, transport, and storage of radioactive materials