Bacterial interactions with uranium: An environmental perspective

The presence of actinides in radioactive wastes is of major concern because of their potential for migration from the waste repositories and long-term contamination of the environment. Studies have been and are being made on inorganic processes affecting the migration of radionuclides from these repositories to the environment but it is becoming increasingly evident that microbial processes are of importance as well. Bacteria interact with uranium through different mechanisms including, biosorption at the cell surface, intracellular accumulation, precipitation, and redox transformations (oxidation/reduction). The present study is intended to give a brief overview of the key processes responsible for the interaction of actinides e.g. uranium with bacterial strains isolated from different extreme environments relevant to radioactive repositories. Fundamental understanding of the interaction of these bacteria with U will be useful for developing appropriate radioactive waste treatments, remediation and long-term management strategies as well as for predicting the microbial impacts on the performance of the radioactive waste repositories.     

Microbial processes as key drivers for metal (im)mobilization along a redox gradient in the saturated zone

Two sites representing different aquifer types, i.e., Dommel (sandy) and Flémalle (gravelly loam) along the Meuse River, have been selected to conduct microcosm experiments. Various conditions ranging from aerobic over nitrate- to sulphate reducing were imposed. For the sandy aquifer, nitrate reducing conditions predominated, which specifically in the presence of a carbon source led to pH increases and enhanced Zn removal. For the calcareous gravelly loam, sulphate reduction was dominant resulting in immobilization of both Zn and Cd. For both aquifer types and almost all redox conditions, higher arsenic concentrations were measured in the groundwater. Analyses of different specific microbial populations by polymerase chain reaction (PCR) revealed the dominance of denitrifiers for the Dommel site, while sulfate reducing bacteria (SRB) were the prevailing population for all redox conditions in the Flémalle samples.     

The potential of lichens as long-term biomonitors of natural and artificial radionuclides

Lichens were used as biomonitors of Chernobyl fallout 137Cs, of cosmogenic 7Be and of radioactive members of the natural uranium and thorium decay chains. Samples were taken from two locations in France, including lichens sampled at different distances of a coal fired power plant and close to a uranium ore processing waste disposal site. All samples were analyzed gamma-spectrometrically after equilibrium concentrations of short-lived isotopes had been attained. Activity concentrations of the members of the uranium and thorium decay chains in Parmelia sulcata sampled 1994 decrease with distance from the plant, whereas in lichens taken at the waste disposal site a decrease with time was observed. Comparison of 7Be activity concentrations measured in lichens with atmospheric deposition rates confirms that P. sulcata can be used as a quantitative biomonitor of radioactive trace substances. Retention half-lives calculated with a simple one-compartment model are 2.6 +/- 1.2 years for cesium, which was detected in all samples even more than a decade after the Chernobyl accident, and of 0.7 (+/- 0.1) to 3.3 (+/- 0.7) years for lead. Consequences of our results for model identifiability and parameter estimation of a two-compartment model are discussed.     

Groundwater chemistry of the Okélobondo uraninite deposit area (Oklo, Gabon): two-dimensional reactive transport modelling

The stability of uranium-bearing minerals in natural environments is of interest to evaluate the feasibility of radioactive waste repositories. The uraninite bodies, UO2(s), in the Oklo district (Gabon) are the result of a natural fission process, which took place 1970 Ma ago. These deposits can be regarded as natural analogues for spent fuel. One of the uraninite bodies, the Okélobondo deposit, is located at a depth of 300 m. Groundwater samples from boreholes located at shallow depths (100-200 m) show neutral to basic pH, anoxic conditions (Eh = 0.10 to -0.05 V) and are saturated with respect to uraninite. In contrast, deeper samples collected in the vicinity of the ore body are oxidising (Eh = 0.32-0.47 V), slightly basic (pH = 7.0-8.5) and undersaturated with respect to uraninite. These oxidising conditions at depth, if present under repository conditions, may affect the stability of uranium oxide. In order to improve our understanding of the observed site geochemistry, the available information on the lithology and groundwater flow was integrated in a reactive transport model. The chemical composition and the pH-Eh values of the water sampled above and in the western side of the Okélobondo deposit can be explained by the interaction of meteoric recharge with pelites, dolomites and sandstones. The dissolution of Fe(II)-silicates and the oxidation of the Fe(II)-aqueous species maintained the pH-Eh distribution along the Fe(2+)-Fe(OH)3(am) equilibrium, with the result that uraninite does not dissolve. This may explain the lower uranium content in the water samples from pelites and dolomites above the Okélobondo deposit. The high Mn/Fe ratio and the high pH-Eh values of the water sampled at depth, close to the Okélobondo deposit, suggest a control by the Mn(2+)-MnOOH(s) equilibrium. This control is attributed to the dissolution of a large rhodochrosite, MnCO3(s), and manganite, MnOOH(s) deposit in the recharge area on the eastern side.     

Intercomparison of reactive transport models applied to UO2 oxidative dissolution and uranium migration

Oxidative dissolution of uranium dioxide (UO(2)) and the subsequent migration of uranium in a subsurface environment and an underground waste disposal have been simulated with reactive transport models. In these systems, hydrogeological and chemical processes are closely entangled and their interdependency has been analyzed in detail, notably with respect to redox reactions, kinetics of mineralogical evolution and hydrodynamic migration of species of interest. Different codes, where among CASTEM, CHEMTRAP and HYTEC, have been used as an intercomparison and verification exercise. Although the agreement between codes is satisfactory, it is shown that the discretization method of the transport equation (i.e. finite elements (FE) versus mixed-hybrid FE and finite differences) and the sequential coupling scheme may lead to systematic discrepancies.     

Intercomparison of reactive transport models applied to UO2 oxidative dissolution and uranium migration

Oxidative dissolution of uranium dioxide (UO₂) and the subsequent migration of uranium in a subsurface environment and an underground waste disposal have been simulated with reactive transport models. In these systems, hydrogeological and chemical processes are closely entangled and their interdependency has been analyzed in detail, notably with respect to redox reactions, kinetics of mineralogical evolution and hydrodynamic migration of species of interest.Different codes, where among CASTEM, CHEMTRAP and HYTEC, have been used as an intercomparison and verification exercise. Although the agreement between codes is satisfactory, it is shown that the discretization method of the transport equation (i.e. finite elements (FE) versus mixed-hybrid FE and finite differences) and the sequential coupling scheme may lead to systematic discrepancies.     

Effect of biogas generation on radon emissions from landfills receiving radium-bearing waste from shale gas development

Dramatic increases in the development of oil and natural gas from shale formations will result in large quantities of drill cuttings, flowback water, and produced water. These organic-rich shale gas formations often contain elevated concentrations of naturally occurring radioactive materials (NORM), such as uranium, thorium, and radium. Production of oil and gas from these formations will also lead to the development of technologically enhanced NORM (TENORM) in production equipment. Disposal of these potentially radium-bearing materials in municipal solid waste (MSW) landfills could release radon to the atmosphere. Risk analyses of disposal of radium-bearing TENORM in MSW landfills sponsored by the Department of Energy did not consider the effect of landfill gas (LFG) generation or LFG control systems on radon emissions. Simulation of radon emissions from landfills with LFG generation indicates that LFG generation can significantly increase radon emissions relative to emissions without LFG generation, where the radon emissions are largely controlled by vapor-phase diffusion. Although the operation of LFG control systems at landfills with radon source materials can result in point-source atmospheric radon plumes, the LFG control systems tend to reduce overall radon emissions by reducing advective gas flow through the landfill surface, and increasing the radon residence time in the subsurface, thus allowing more time for radon to decay. In some of the disposal scenarios considered, the radon flux from the landfill and off-site atmospheric activities exceed levels that would be allowed for radon emissions from uranium mill tailings.

Implications: Increased development of hydrocarbons from organic-rich shale formations has raised public concern that wastes from these activities containing naturally occurring radioactive materials, particularly radium, may be disposed in municipal solid waste landfills and endanger public health by releasing radon to the atmosphere. This paper analyses the processes by which radon may be emitted from a landfill to the atmosphere. The analyses indicate that landfill gas generation can significantly increase radon emissions, but that the actual level of radon emissions depend on the place of the waste, construction of the landfill cover, and nature of the landfill gas control system.     

Uranium transport around the reactor zone at Bangombé and Okélobondo (Oklo): examples of hydrogeological and geochemical model integration and data evaluation

The sites at Bangombé and Okélobondo (Oklo) in Gabon provide a unique opportunity to study the behaviour of products from natural nuclear reactions in the vicinity of reactor zones which were active around two billion years ago. The Commission of the European Communities initiated the Oklo Natural Analogue Programme. One of the principal aims was to study indications of present time migration of elements from the reactor zones under ambient conditions. The hydrogeological and hydrochemical data from the Oklo sites were modelled in order to better understand the geochemical behaviour of radionuclides in the natural system, by using independent models and by comparing the modelling outcome. Two modelling approaches were used: M3 code (hydrochemical mixing and mass balance model), developed by the Swedish Nuclear Fuel and Waste Management Company (SKB) and HYTEC (reactive transport model) developed by Ecole des Mines de Paris. Two different reactor zones were studied: Bangombé, a shallow site, the reactor being at 11 m depth, and OK84 at Okélobondo, situated at about 450 m depth, more comparable with a real repository location. This allowed the validation of modelling tools in two different sedimentary environments: one shallow, with a more homogeneous layering situated in an area of meteoric alteration, and the other offering the opportunity to study radionuclide migration from the reaction zone over a distance of 450 m through very heterogeneous sedimentary layers. The modeling results indicate that the chemical reactions retarding radionuclide transport are very different at the two sites. At Bangombé, the decomposition of organic material consumes oxygen and at Okélobondo the oxygen is consumed by inorganic reactions resulting, in both cases, in uranium retardation. Both modelling approaches (statistic with M3 code and deterministic with HYTEC code) could describe this situation. The goal of this exercise is to test codes which can help to describe and understand the processes taking place at the sites, validate the models with in situ data, and thus build confidence in the tools used for future site characterization. Ultimately, this allows identifying and selecting processes and parameters that can be used as input into repository performance assessment calculations and modelling exercises.     

Transport and fate of organic wastes in groundwater at the Stringfellow hazardous waste disposal site, southern California

In January 1999, wastewater influent and effluent from the pretreatment plant at the Stringfellow hazardous waste disposal site were sampled along with groundwater at six locations along the groundwater contaminant plume. The objectives of this sampling and study were to identify at the compound class level the unidentified 40-60% of wastewater organic contaminants, and to determine what organic compound classes were being removed by the wastewater pretreatment plant, and what organic compound classes persisted during subsurface waste migration. The unidentified organic wastes are primarily chlorinated aromatic sulfonic acids derived from wastes from DDT manufacture. Trace amounts of EDTA and NTA organic complexing agents were discovered along with carboxylate metabolites of the common alkylphenolpolyethoxylate plasticizers and nonionic surfactants. The wastewater pretreatment plant removed most of the aromatic chlorinated sulfonic acids that have hydrophobic neutral properties, but the p-chlorobenzene-sulfonic acid which is the primary waste constituent passed through the pretreatment plant and was discharged in the treated wastewaters transported to an industrial sewer. During migration in groundwater, p-chlorobenzenesulfonic acid is removed by natural remediation processes. Wastewater organic contaminants have decreased 3- to 45-fold in the groundwater from 1985 to 1999 as a result of site remediation and natural remediation processes. The chlorinated aromatic sulfonic acids with hydrophobic neutral properties persist and have migrated into groundwater that underlies the adjacent residential community.     

Kinetic modeling of microbially-driven redox chemistry of radionuclides in subsurface environments: coupling transport, microbial metabolism and geochemistry

Microbial reactions play an important role in regulating pore water chemistry as well as secondary mineral distribution in many subsurface systems and, therefore, may directly impact radionuclide migration in those systems. This paper presents a general modeling approach to couple microbial metabolism, redox chemistry, and radionuclide transport in a subsurface environment. To account for the likely achievement of quasi-steady state biomass accumulations in subsurface environments, a modification to the traditional microbial growth kinetic equation is proposed. The conditions for using biogeochemical models with or without an explicit representation of biomass growth are clarified. Based on the general approach proposed in this paper, the couplings of uranium reactions with biogeochemical processes are incorporated into computer code BIORXNTRN Version 2.0. The code is then used to simulate a subsurface contaminant migration scenario, in which a water flow containing both uranium and a complexing organic ligand is recharged into an oxic carbonate aquifer. The model simulation shows that Mn and Fe oxyhydroxides may vary significantly along a flow path. The simulation also shows that uranium(VI) can be reduced and therefore immobilized in the anoxic zone created by microbial degradation.     

Removal of chromium from synthetic plating waste by zero-valent iron and sulfate-reducing bacteria.

Experiments were conducted to evaluate the potential of zero-valent iron and sulfate-reducing bacteria (SRB) for reduction and removal of chromium from synthetic electroplating waste. The zero-valent iron shows promising results as a reductant of hexavalent chromium (Cr+6) to trivalent chromium (Cr+3), capable of 100% reduction. The required iron concentration was a function of chromium concentration in the waste stream. Removal of Cr+3 by adsorption or precipitation on iron leads to complete removal of chromium from the waste and was a slower process than the reduction of Cr+6. Presence SRB in a completely mixed batch reactor inhibited the reduction of Cr+6. In a fixed-bed column reactor, SRB enhanced chromium removal and showed promising results for the treatment of wastes with low chromium concentrations. It is proposed that, for waste with high chromium concentration, zero-valent iron is an efficient reductant and can be used for reduction of Cr+6. For low chromium concentrations, a SRB augmented zero-valent iron and sand column is capable of removing chromium completely.     

Radioactive waste forms stabilized by ChemChar gasification: characterization and leaching behavior of cerium, thorium, protactinium, uranium, and neptunium

The uses of a thermally reductive gasification process in conjunction with vitrification and cementation for the long-term disposal of low level radioactive materials have been investigated. gamma-ray spectroscopy was used for analysis of carrier-free protactinium-233 and neptunium-239 and a stoichiometric amount of cerium (observed cerium-141) subsequent to gasification and leaching, up to 48 days. High resolution ICP-MS was used to analyze the cerium, thorium, and uranium from 46 to 438 days of leaching. Leaching procedures followed the guidance of ASTM Procedure C 1220-92, Standard Test Method for Static Leaching of Monolithic Waste Forms for Disposal of Radioactive Waste. The combination of the thermally reductive pretreatment, vitrification and cementation produced a highly non-leachable form suitable for long-term disposal of cerium, thorium, protactinium, uranium, and neptunium.     

Large-scale modeling of reactive solute transport in fracture zones of granitic bedrocks

Final disposal of high-level radioactive waste in deep repositories located in fractured granite formations is being considered by several countries. The assessment of the safety of such repositories requires using numerical models of groundwater flow, solute transport and chemical processes. These models are being developed from data and knowledge gained from in situ experiments such as the Redox Zone Experiment carried out at the underground laboratory of Äspö in Sweden. This experiment aimed at evaluating the effects of the construction of the access tunnel on the hydrogeological and hydrochemical conditions of a fracture zone intersected by the tunnel. Most chemical species showed dilution trends except for bicarbonate and sulphate which unexpectedly increased with time. Molinero and Samper [Molinero, J. and Samper, J. Groundwater flow and solute transport in fracture zones: an improved model for a large-scale field experiment at Äspö (Sweden). J. Hydraul. Res., 42, Extra Issue, 157–172] presented a two-dimensional water flow and solute transport finite element model which reproduced measured drawdowns and dilution curves of conservative species. Here we extend their model by using a reactive transport which accounts for aqueous complexation, acid–base, redox processes, dissolution–precipitation of calcite, quartz, hematite and pyrite, and cation exchange between Na⁺ and Ca²⁺. The model provides field-scale estimates of cation exchange capacity of the fracture zone and redox potential of groundwater recharge. It serves also to identify the mineral phases controlling the solubility of iron. In addition, the model is useful to test the relevance of several geochemical processes. Model results rule out calcite dissolution as the process causing the increase in bicarbonate concentration and reject the following possible sources of sulphate: (1) pyrite dissolution, (2) leaching of alkaline sulphate-rich waters from a nearby rock landfill and (3) dissolution of iron monosulphides contained in Baltic seafloor sediments. Based on these results, microbially mediated processes are postulated as the most likely hypothesis to explain the measured increase of dissolved bicarbonates and sulphates after tunnel construction.     

Microbial community dynamics in uranium contaminated subsurface sediments under biostimulated conditions with high nitrate and nickel pressure

BACKGROUND, AIM, AND SCOPE: The subsurface at the Oak Ridge Field Research Center represents an extreme and diverse geochemical environment that places different stresses on the endogenous microbial communities, including low pH, elevated nitrate concentrations, and the occurrence of heavy metals and radionuclides, including hexavalent uranium [U(VI)]. The in situ immobilization of U(VI) in the aquifer can be achieved through microbial reduction to relatively insoluble U(IV). However, a high redox potential due to the presence of nitrate and the toxicity of heavy metals will impede this process. Our aim is to test biostimulation of the endogenous microbial communities to improve nitrate reduction and subsequent U(VI) reduction under conditions of elevated heavy metals. MATERIALS AND METHODS: Column experiments were used to test the possibility of using biostimulation via the addition of ethanol as a carbon source to improve nitrate reduction in the presence of elevated aqueous nickel. We subsequently analyzed the composition of the microbial communities that became established and their potential for U(VI) reduction and its in situ immobilization. RESULTS: Phylogenetic analysis revealed that the microbial population changed from heavy metal sensitive members of the actinobacteria, alpha- and gamma-proteobacteria to a community dominated by heavy metal resistant (nickel, cadmium, zinc, and cobalt resistant), nitrate reducing beta- and gamma-proteobacteria, and sulfate reducing Clostridiaceae. Coincidentally, synchrotron X-ray absorption spectroscopy analyses indicated that the resulting redox conditions favored U(VI) reduction transformation to insoluble U(IV) species associated with soil minerals and biomass. DISCUSSION: This study shows that the necessary genetic information to adapt to the implemented nickel stress resides in the endogenous microbial population present at the Oak Ridge FRC site, which changed from a community generally found under oligotrophic conditions to a community able to withstand the stress imposed by heavy metals, while efficiently reducing nitrate as electron donor. Once nitrate was reduced efficient reduction and in situ immobilization of uranium was observed. CONCLUSIONS: This study provides evidence that stimulating the metabolism of the endogenous bacterial population at the Oak Ridge FRC site by adding ethanol, a suitable carbon source, results in efficient nitrate reduction under conditions of elevated nickel, and a decrease of the redox potential such that sulfate and iron reducing bacteria are able to thrive and create conditions favorable for the reduction and in situ immobilization of uranium. Since we have found that the remediation potential resides within the endogenous microbial community, we believe it will be feasible to conduct field tests. RECOMMENDATIONS AND PERSPECTIVES: Biostimulation of endogenous bacteria provides an efficient tool for the successful in situ remediation of mixed-waste sites, particularly those co-contaminated with heavy metals, nitrate and radionuclides, as found in the United States and other countries as environmental legacies of the nuclear age.     

Geochemical evaluation of different groundwater-host rock systems for radioactive waste disposal

The geochemical suitability of a deep bedrock repository for radioactive waste disposal is determined by the composition of geomatrix and groundwater. Both influence radionuclide solubility, chemical buffer capacity and radionuclide retention. They also determine the chemical compatibility of waste forms, containers and backfill materials. Evaluation of different groundwater-host rock systems is performed by modeling the geochemical environments and the resulting radionuclide concentrations. In order to demonstrate the evaluation method, model calculations are applied to data sets available for various geological formations such as granite, clay and rocksalt. The saturation state of the groundwater-geomatrix system is found to be fundamental for the evaluation process. Hence, calculations are performed to determine if groundwater is in equilibrium with mineral phases of the geological formation. In addition, corrosion of waste forms in different groundwater is examined by means of reaction path modeling. The corrosion reactions change the solution compositions and pH, resulting in significant changes of radionuclide solubilities. The results demonstrate that geochemical modeling of saturation state and compatibility of the host formation environment with the radioactive waste proves to be a feasible tool for evaluation of various sites considered as deep underground repositories.     

Application of in situ biosparging to remediate a petroleum-hydrocarbon spill site: field and microbial evaluation

In this study, a full-scale biosparging investigation was conducted at a petroleum-hydrocarbon spill site. Field results reveal that natural attenuation was the main cause of the decrease in major contaminants [benzene, toluene, ethylbenzene, and xylenes (BTEX)] concentrations in groundwater before the operation of biosparging system. Evidence of the occurrence of natural attenuation within the BTEX plume includes: (1) decrease of DO, nitrate, sulfate, and redox potential, (2) production of dissolved ferrous iron, sulfide, methane, and CO(2), (3) decreased BTEX concentrations along the transport path, (4) increased microbial populations, and (5) limited spreading of the BTEX plume. Field results also reveal that the operation of biosparging caused the shifting of anaerobic conditions inside the plume to aerobic conditions. This variation can be confirmed by the following field observations inside the plume due to the biosparging process: (1) increase in DO, redox potential, nitrate, and sulfate, (2) decrease dissolved ferrous iron, sulfide, and methane, (3) increased total cultivable heterotrophs, and (4) decreased total cultivable anaerobes as well as methanogens. Results of polymerase chain reaction, denaturing gradient gel electrophoresis, and nucleotide sequence analysis reveal that three BTEX biodegraders (Candidauts magnetobacterium, Flavobacteriales bacterium, and Bacteroidetes bacterium) might exist at this site. Results show that more than 70% of BTEX has been removed through the biosparging system within a 10-month remedial period at an averaged groundwater temperature of 18 degrees C. This indicates that biosparging is a promising technology to remediate BTEX contaminated groundwater.     

Geochemical evaluation of different groundwater–host rock systems for radioactive waste disposal

The geochemical suitability of a deep bedrock repository for radioactive waste disposal is determined by the composition of geomatrix and groundwater. Both influence radionuclide solubility, chemical buffer capacity and radionuclide retention. They also determine the chemical compatibility of waste forms, containers and backfill materials. Evaluation of different groundwater–host rock systems is performed by modeling the geochemical environments and the resulting radionuclide concentrations. In order to demonstrate the evaluation method, model calculations are applied to data sets available for various geological formations such as granite, clay and rocksalt.The saturation state of the groundwater–geomatrix system is found to be fundamental for the evaluation process. Hence, calculations are performed to determine if groundwater is in equilibrium with mineral phases of the geological formation. In addition, corrosion of waste forms in different groundwater is examined by means of reaction path modeling. The corrosion reactions change the solution compositions and pH, resulting in significant changes of radionuclide solubilities. The results demonstrate that geochemical modeling of saturation state and compatibility of the host formation environment with the radioactive waste proves to be a feasible tool for evaluation of various sites considered as deep underground repositories.     

U-series disequilibria in a groundwater flow route as an indicator of uranium migration processes

U-series data relating to groundwater, fracture coatings and the adjoining rock matrix in a groundwater flow system at the Palmottu natural analogue site was examined. The aim was to obtain an experimental reference for migration modelling in a transport section defined within the flow system. The U-series reference obtained turned out to be a very useful tool for fine tuning the flow route and for migration mechanism considerations. The U-series data are well in line with other interpretations of the migration system.     

Pseudomonas fluorescens dynamics in the soil surface to subsurface transect

Microbial displacement in the soil is an important process for bioremediation and dispersal of wastewater pathogens. We evaluated cell movement in surface and subsurface red-yellow podzolic soil driven by advection and microbial motility and also survival of a microbial population at high pressure as is prevalent in deep soil layers. Pseudomonas fluorescens Br 12, resistant to rifampycin and kanamycin, was used as a model organism traceable in non-sterile soil. Our results showed that more than 40% of the P. fluorescens population survived under high pressure, and that microbial motility was not a major factor for its displacement in the soil. Cells were adsorbed in similar amounts to surface and subsurface soils, but more viable cells were present in the leachate of surface than in subsurface soils. The nature of this unexpected cell binding to the subsurface soil was studied by EPR, Mossbauer, NMR, and infrared techniques, suggesting iron had a weak interaction with microbes in soil. P. fluorescens movement in soil resulted mainly from convection forces rather than microbial motility. The transport of this bacterium along the transept toward groundwater encountered restricted viability, although it survived under high pressure conditions simulating those in deep soil layers.