Pilot-scale in situ bioremediation of uranium in a highly contaminated aquifer. 1. Conditioning of a treatment zone.

To evaluate the potential for in situ bioremediation of U(VI) to sparingly soluble U(IV), we constructed a pilot test facility at Area 3 of the U.S. Department of Energy Natural and Accelerated Bioremediation Research (NABIR) Field Research Center (FRC) in Oak Ridge, TN. The facility is adjacent to the former S-3 Ponds which received trillions of liters of acidic plating wastes. High levels of uranium are present, with up to 800 mg kg(-1) in the soil and 84-210 microM in the groundwater. Ambient groundwater has a highly buffered pH of approximately 3.4 and high levels of aluminum (12-13 mM), calcium (22-25 mM), and nitrate (80-160 mM). Adjusting the pH of groundwater to approximately 5 within the aquifer would deposit extensive aluminum hydroxide precipitate. Calcium is present in the groundwater at levels that inhibit U(VI) reduction, but its removal by injection of a high pH solution would generate clogging precipitate. Nitrate also inhibits U(VI) reduction and is present at such high concentrations that its removal by in situ denitrification would generate large amounts of N2 gas and biomass. To establish and maintain hydraulic control, we installed a four well recirculation system parallel to geologic strike, with an inner loop nested within an outer loop. For monitoring, we drilled three boreholes perpendicular to strike across the inner loop and installed multilevel sampling tubes within them. A tracer pulse with clean water established travel times and connectivity between wells and enabled the assessment of contaminant release from the soil matrix. Subsequently, a highly conductive region of the subsurface was prepared for biostimulation by removing clogging agents and inhibitors and increasing pH. For 2 months, groundwater was pumped from the hydraulically conductive zone; treated to remove aluminum, calcium, and nitrate, and supplemented with tap water; adjusted to pH 4.3-4.5; then returned to the hydraulically conductive zone. This protocol removed most of the aqueous aluminum and calcium. The pH of the injected treated water was then increased to 6.0-6.3. With additional flushing, the pH of the extracted water gradually increased to 5.5-6.0, and nitrate concentrations fell to 0.5-1.0 mM. These conditions were judged suitable for biostimulation. In a companion paper (Wu et al., Environ. Sci. Technol. 2006, 40, 3978-3987), we describe the effects of ethanol addition on in situ denitrification and U(VI) reduction and immobilization.

Pilot-scale in situ bioremedation of uranium in a highly contaminated aquifer. 2. Reduction of u(VI) and geochemical control of u(VI) bioavailability.

In situ microbial reduction of soluble U(VI) to sparingly soluble U(IV) was evaluated at the site of the former S-3 Ponds in Area 3 of the U.S. Department of Energy Natural and Accelerated Bioremediation Research Field Research Center, Oak Ridge, TN. After establishing conditions favorable for bioremediation (Wu, et al. Environ. Sci. Technol. 2006, 40, 3988-3995), intermittent additions of ethanol were initiated within the conditioned inner loop of a nested well recirculation system. These additions initially stimulated denitrification of matrix-entrapped nitrate, but after 2 months, aqueous U levels fell from 5 to approximately 1 microM and sulfate reduction ensued. Continued additions sustained U(VI) reduction over 13 months. X-ray near-edge absorption spectroscopy (XANES) confirmed U(VI) reduction to U(IV) within the inner loop wells, with up to 51%, 35%, and 28% solid-phase U(IV) in sediment samples from the injection well, a monitoring well, and the extraction well, respectively. Microbial analyses confirmed the presence of denitrifying, sulfate-reducing, and iron-reducing bacteria in groundwater and sediments. System pH was generally maintained at less than 6.2 with low bicarbonate level (0.75-1.5 mM) and residual sulfate to suppress methanogenesis and minimize uranium mobilization. The bioavailability of sorbed U(VI) was manipulated by addition of low-level carbonate (< 5 mM) followed by ethanol (1-1.5 mM). Addition of low levels of carbonate increased the concentration of aqueous U, indicating an increased rate of U desorption due to formation of uranyl carbonate complexes. Upon ethanol addition, aqueous U(VI) levels fell, indicating that the rate of microbial reduction exceeded the rate of desorption. Sulfate levels simultaneously decreased, with a corresponding increase in sulfide. When ethanol addition ended but carbonate addition continued, soluble U levels increased, indicating faster desorption than reduction. When bicarbonate addition stopped, aqueous U levels decreased, indicating adsorption to sediments. Changes in the sequence of carbonate and ethanol addition confirmed that carbonate-controlled desorption increased bioavailability of U(VI) for reduction.

Stimulating the anaerobic biodegradation of explosives by the addition of hydrogen or electron donors that produce hydrogen.

The anaerobic biodegradation of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), and 2,4,6-trinitrotoluene (TNT) by a methanogenic mixed culture was investigated. Microcosms containing a basal medium and the mixed culture were amended with ethanol, propylene glycol (PG), butyrate or hydrogen gas as the electron donor and a mixture of TNT (50 microM), RDX (25 microM), and HMX (8 microM). After 29 days TNT and RDX were completely transformed to unidentified endproducts in the bottles amended with ethanol, hydrogen, or PG, while 53%, 40%, and 22% of the HMX was transformed, respectively. There was no loss of RDX or HMX in the electron donor unamended control bottles. The ethanol and PG were transformed to near stoichiometric amounts of acetate and propionate, suggesting the immediate electron donor supporting the transformation of the explosives was the H2 evolved during the metabolism of the parent substrate. Our findings suggest that the addition of H2 or electron donors that produce H2 may be a useful strategy for enhancing the anaerobic biodegradation of explosives in contaminated groundwater and soils.

Anaerobic treatment of pinkwater in a fluidized bed reactor containing GAC.

Pinkwater is generated during the handling and demilitarization of conventional explosives. This listed hazardous waste contains dissolved trinitrotoluene (TNT) and cyclo trimethylene trinitramine (RDX), as well as some by-products. It represents the largest quantity of hazardous waste generated by the operations support command, and its treatment produces a by-product hazardous waste--spent granular activated carbon (GAC). Anaerobic treatment in a fluidized bed reactor (FBR) containing GAC is an emerging technology for organic compounds resistant to aerobic biological treatment. Bench scale batch studies using an anaerobic consortium of bacteria fed ethanol as the sole electron donor demonstrated the transformation of TNT to triaminotoluene (TAT), which then degrades to undetectable end products. RDX is sequentially degraded to nitroso-, dinitroso-, trinitroso- and hydroxylaminodinitroso-RDX before the triazine ring is presumably cleaved, forming methanol and formaldehyde as major end products. The bacterial members of the anaerobic consortia are typically found in sludge digesters at municipal or industrial wastewater treatment plants. The results of a pilot scale evaluation of this process that was conducted at McAlester Army Ammunition Plant (MCAAP, OK) over a 1 year period are reported in this paper. The pilot test experienced wide fluctuations in influent concentrations, representative of true field conditions. The FBR was a 20 in. (51 cm) diameter column with an overall height of 15 ft (4.9 m) and a bed of GAC occupying 11 ft (3.4m). Water was recirculated through the column continuously at 30 gpm (114 l/min) to keep the GAC fluidized, and pinkwater for treatment was pumped into the recirculation line. Several flowrates were evaluated to determine the proper mass loading rate (mass of TNT and RDX per reactor volume per time, kg/m(3) per day) which the reactor could handle while meeting the discharge limitations. Based on the tests performed, a 1 gpm (3.785 l/min) rate in the 188 gal (710 l) volume of the fluidized GAC bed was determined to consistently meet the discharge requirements. This information was used to develop a cost estimate for a system capable of treating the total effluent currently produced at MCAAP. The cost of installing and operating this system was compared to the cost of GAC adsorption for MCAAP at current pinkwater generation rates. The GAC-FBR system had an annual operating cost of approximately US$ 19K, compared to US$ 71 K annually for GAC adsorption. When including the amortization of the capital equipment required for the GAC-FBR, the payback period for installation of this new process was estimated at 3.7 years.