Mechanisms for the Reduction of Actinides and Tc(VII) in Geobacter sulfurreducens

The mechanism of the reduction of U(VI) and Cr(VI) has now been studied in detail. Cr(VI) is reduced by one-electron transfer reactions to Cr(III), via a cell-bound Cr(V) intermediate identified by EPR spectroscopy. Studies with a cytochrome c7 mutant demonstrate that the electron transfer chain includes this protein which may be the terminal reductase for Cr(VI). Potential mechanisms of inhibition of Cr(III) precipitation, involving complex formation with organic acids commonly used as electron donors for metal reduction in the subsurface have also been identified. We have also initiated a collaboration with computational chemists led by Prof Ian Hillier in Manchester, to model metal binding to cytochrome c7, and subsequent electron transfer from the enzyme to the metal quantum mechanically

Novel Imaging Techniques, Integrated with Mineralogical, Geochemical and Microbiological Characterization to Determine the Biogeochemical Controls....

Tc(VII) will be reduced and precipitated in FRC sediments under anaerobic conditions in batch experiments (progressive microcosms). The complementary microcosm experiments using low pH/nigh nitrate sediments from 3 (near FW 009) are imminent, with the sediment cores already shipped to Manchester. HYPOTHESIS 2. Tc(VII) reduction and precipitation can be visualized in discrete biogeochemical zones in sediment columns using 99mTc and a gamma-camera. Preliminary experiments testing the use of 99mTc as a radiotracer to address hypotheses 2 and 3 have suggested that the 99mTc associates with Fe(II)-bearing sediments in microcosms and stratified columns containing FRC sediments. Initial proof of concept microcosms containing Fe(II)-bearing, microbially-reduced FRC sediments were spiked with 99mTc and imaged using a gamma-camera. In comparison with oxic controls, 99mTc was significantly partitioned in the solid phase in Fe(III)-reducing sediments in batch experiments. Column experiments using FRC background area soil with stratified biogeochemical zones after stimulation of anaerobic processes through nutrient supplementation, suggested that 99mTc transport was retarded through areas of Fe(III) reduction. HYPOTHESIS 3. Sediment-bound reduced 99mTc can be solubilized by perturbations including oxidation coupled to biological nitrate reduction, and mobilization visualized in real-time using a gamma-camera. Significant progress has been made focusing on the impact of nitrate on the biogeochemical behavior of technetium. Additions of 100 mM nitrate to FRC sediment microcosms, which could potentially compete for electrons during metal reduction, inhibited the reduction of both Fe(III) and Tc(VII) completely. Experiments have also addressed the impact of high nitrate concentrations on Fe(II) and Tc(IV) in pre-reduced sediments, showing no significant resolubilization of Tc with the addition of 25 mM nitrate. A parallel set of experiments addressing the impact of aerobic conditions on the stability/solubility of Fe(II) and Tc(IV), found 80 % resolubilization of the Tc. Column experiments exploring this behavior are being planned. HYPOTHESIS 4 The mobility of 99mTc in the sediment columns can be modeled using a coupled speciation and transport code. Microbiological and geochemical characterization of the column experiments is ongoing and transport and geochemical modeling experiments are being planned

Novel imaging techniques, integrated with mineralogical, geochemical and microbiological characterizations to determine the biogeochemical controls on technetium mobility in FRC sediments

The objective of this research program was to take a highly multidisciplinary approach to define the biogeochemical factors that control technetium (Tc) mobility in FRC sediments. The aim was to use batch and column studies to probe the biogeochemical conditions that control the mobility of Tc at the FRC. Background sediment samples from Area 2 (pH 6.5, low nitrate, low {sup 99}Tc) and Area 3 (pH 3.5, high nitrate, relatively high {sup 99}Tc) of the FRC were selected (http://www.esd.ornl.gov/nabirfrc). For the batch experiments, sediments were mixed with simulated groundwater, modeled on chemical constituents of FRC waters and supplemented with {sup 99}Tc(VII), both with and without added electron donor (acetate). The solubility of the Tc was monitored, alongside other biogeochemical markers (nitrate, nitrite, Fe(II), sulfate, acetate, pH, Eh) as the 'microcosms' aged. At key points, the microbial communities were also profiled using both cultivation-dependent and molecular techniques, and results correlated with the geochemical conditions in the sediments. The mineral phases present in the sediments were also characterized, and the solid phase associations of the Tc determined using sequential extraction and synchrotron techniques. In addition to the batch sediment experiments, where discrete microbial communities with the potential to reduce and precipitate {sup 99}Tc will be separated in time, we also developed column experiments where biogeochemical processes were spatially separated. Experiments were conducted both with and without amendments proposed to stimulate radionuclide immobilization (e.g. the addition of acetate as an electron donor for metal reduction), and were also planned with and without competing anions at high concentration (e.g. nitrate, with columns containing Area 3 sediments). When the columns had stabilized, as determined by chemical analysis of the effluents, we used a spike of the short-lived gamma emitter {sup 99m}Tc (50-200 MBq; half life 6 hours) and its mobility was monitored using a {gamma}-camera. Incorporation of low concentrations of the long-lived 99Tc gave a tracer that can be followed by scintillation counting, should the metastable form of the radionuclide decay to below detection limits before the end of the experiment (complete immobilization or loss of the Tc from the column). After the Tc was reduced and immobilized, or passed through the system, the columns were dismantled carefully in an anaerobic cabinet and the pore water geochemistry and mineralogy of the columns profiled. Microbial community analysis was determined, again using molecular and culture-dependent techniques. Experimental results were also modeled using an established coupled speciation and transport code, to develop a predictive tool for the mobility of Tc in FRC sediments. From this multidisciplinary approach, we hoped to obtain detailed information on the microorganisms that control the biogeochemical cycling of key elements at the FRC, and we would also be able to determine the key factors that control the mobility of Tc at environmentally relevant concentrations at this site

The Impact of Gamma Radiation on Sediment Microbial Processes

Microbial communities have the potential to control the biogeochemical fate of some radionuclides in contaminated land scenarios or in the vicinity of a geological repository for radioactive waste. However, there have been few studies of ionizing radiation effects on microbial communities in sediment systems. Here, acetate and lactate amended sediment microcosms irradiated with gamma radiation at 0.5 or 30 Gy h(−1) for 8 weeks all displayed NO(3)(−) and Fe(III) reduction, although the rate of Fe(III) reduction was decreased in 30-Gy h(−1) treatments. These systems were dominated by fermentation processes. Pyrosequencing indicated that the 30-Gy h(−1) treatment resulted in a community dominated by two Clostridial species. In systems containing no added electron donor, irradiation at either dose rate did not restrict NO(3)(−), Fe(III), or SO(4)(2−) reduction. Rather, Fe(III) reduction was stimulated in the 0.5-Gy h(−1)-treated systems. In irradiated systems, there was a relative increase in the proportion of bacteria capable of Fe(III) reduction, with Geothrix fermentans and Geobacter sp. identified in the 0.5-Gy h(−1) and 30-Gy h(−1) treatments, respectively. These results indicate that biogeochemical processes will likely not be restricted by dose rates in such environments, and electron accepting processes may even be stimulated by radiation

Treatment of Alkaline Cr(VI)-Contaminated Leachate with an Alkaliphilic Metal-Reducing Bacterium

Chromium in its toxic Cr(VI) valence state is a common contaminant particularly associated with alkaline environments. A well-publicized case of this occurred in Glasgow, United Kingdom, where poorly controlled disposal of a cementitious industrial by-product, chromite ore processing residue (COPR), has resulted in extensive contamination by Cr(VI)-contaminated alkaline leachates. In the search for viable bioremediation treatments for Cr(VI), a variety of bacteria that are capable of reduction of the toxic and highly soluble Cr(VI) to the relatively nontoxic and less mobile Cr(III) oxidation state, predominantly under circumneutral pH conditions, have been isolated. Recently, however, alkaliphilic bacteria that have the potential to reduce Cr(VI) under alkaline conditions have been identified. This study focuses on the application of a metal-reducing bacterium to the remediation of alkaline Cr(VI)-contaminated leachates from COPR. This bacterium, belonging to the Halomonas genus, was found to exhibit growth concomitant to Cr(VI) reduction under alkaline conditions (pH 10). Bacterial cells were able to rapidly remove high concentrations of aqueous Cr(VI) (2.5 mM) under anaerobic conditions, up to a starting pH of 11. Cr(VI) reduction rates were controlled by pH, with slower removal observed at pH 11, compared to pH 10, while no removal was observed at pH 12. The reduction of aqueous Cr(VI) resulted in the precipitation of Cr(III) biominerals, which were characterized using transmission electron microscopy and energy-dispersive X-ray analysis (TEM-EDX) and X-ray photoelectron spectroscopy (XPS). The effectiveness of this haloalkaliphilic bacterium for Cr(VI) reduction at high pH suggests potential for its use as an in situ treatment of COPR and other alkaline Cr(VI)-contaminated environments

454 pyrosequencing assessment of biodegradative bacteria from thermal hydrolysis processes

Anaerobic treatment process is a cost-effective method for treating organic wastes, since the biogas formed can be used for heat/electricity production and the digester residues can be recycled for other applications. An innovative use of the digestate could be as biodegradative and methanogenic inoculum for the stimulation of methane production in gas-producing or depleted wells. The microbial communities involved in the biodegradation of petrochemical waste are similar to the indigenous microorganisms typically found in unconventional basins. These communities also follow the same cascade of reactions: from the initial breakdown of complex molecules to the production of intermediate compounds used by methanogens. This study carried out a culture-independent assessment of the bacterial community composition of a digestate from the Bran Sands Advanced Digestion Facility (Middleborough, UK) and compared the results with the microbial populations found in unconventional gas basins. The 454 pyrosequencing analyses revealed a bacterial community dominated by Thermotogae, Bacteroidia, Clostridia and Synergistia, which are typically found in unconventional gas systems. The classification of nucleotide sequence reads and assembled contigs revealed a genetic profile characteristic for an anaerobic microbial consortium running fermentative metabolic pathways. The assignment of numerous sequences was related to hydrocarbon decomposition and digestion of cellulosic material, which indicates that the bacterial community is engaged in hydrolysis of plant-derived material. The bacterial community composition suggest that the effluent of the digester can be used as a biodegradative inoculum for the stimulation of methane generation in unconventional wells, where events of microbial methanogenesis have been previously observed

Rapid and Deep Remission Induced by Blinatumomab for CD19-Positive Chronic Myeloid Leukemia in Lymphoid Blast Phase

In summary, we show rapid and deep remission induced by blinatumomab in CD19(+) blast phase CML. Clinicians may consider the use of bispecific T-cell engager therapy as a bridge to transplant. Additional studies are needed before expanding the US Food and Drug Administration indication of blinatumomab to include lymphoid blast phase CML