Arokiasamy J. Francis

XPS and XANES Studies of Uranium Reduction by Clostridium sp.

Speciation of uranium in cultures of Clostridium sp. by X-ray absorption near-edge spectroscopy (XANES) at the National Synchrotron Light Source and by X-ray photoelectron spectroscopy (XPS) showed that U(VI) was reduced to U(IV). In addition to U(IV), a lower oxidation state of uranium, most probably U(III), was detected by XANES in the bacterial cultures. Reduction of uranium occurred only in the presence of growing or resting cells. Organic acid metabolites, the extracellular components of the culture medium, and heat-killed cells failed to reduce uranium under anaerobic conditions. The addition of uranyl acetate or uranyl nitrate (>210[mu]M U) to the culture medium retarded the growth of the bacteria as evidenced by an increase in the lag period before resumption of growth, by decreases in turbidity, and in the total production of gas and organic acid metabolites. These results show that uranium in wastes can be stabilized by the action of anaerobic bacteria. 31 refs., 2 figs., 3 tabs.

Biotransformation of uranium and other actinides in radioactive wastes

Microorganisms affect the solubility, bioavailability, and mobility of actinides in radioactive wastes. Under appropriate conditions, actinides are solubilized or stabilized by the direct enzymatic or indirect nonenzymatic actions of microorganisms. Biotransformation of various forms of uranium (ionic, inorganic, and organic complexes) by aerobic and anaerobic microorganisms has been extensively studied, whereas limited information is available on other important actinides (Th, Np, Pu, and Am). Fundamental information on the mechanisms of biotransformation of actinides by microbes under various environmental conditions will be useful in predicting the long-term performance of waste repositories and in developing strategies for waste management and remediation of contaminated sites.

Uranium association with halophilic and non-halophilic bacteria and archaea

Summary We determined the association of uranium with bacteria isolated from the Waste Isolation Pilot Plant (WIPP), Carlsbad, New Mexico, and compared this with known strains of halophilic and non-halophilic bacteria and archaea. Examination of the cultures by transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDS) showed uranium accumulation extracellularly and/or intracellularly to a varying degree. In Pseudomonas fluorescens and Bacillus subtilis uranium was associated with the cell surface and in the latter it was present as irregularly shaped grains. In Halobacterium halobium, the only archeon studied here, uranium was present as dense deposits and with Haloanaerobium praevalens as spikey deposits. Halomonas sp. isolated from the WIPP site accumulated uranium both extracellularly on the cell surface and intracellularly as electron-dense discrete granules. Extended X-ray absorption fine structure (EXAFS) analysis of uranium with the halophilic and non-halophilic bacteria and archaea showed that the uranium present in whole cells was bonded to an average of 2.4±0.7 phosphoryl groups at a distance of 3.65±0.03 Å. Comparison of whole cells of Halomonas sp. with the cell wall fragments of lysed cells showed the presence of a uranium bidentate complex at 2.91±0.03 Å with the carboxylate group on the cell wall, and uranyl hydroxide with U-U interaction at 3.71±0.03 Å due to adsorption or precipitation reactions; no U-P interaction was observed. Addition of uranium to the cell lysate of Halomonas sp. resulted in the precipitation of uranium due to the inorganic phosphate produced by the cells. These results show that the phosphates released from bacteria bind a significant amount of uranium. However, the bacterially immobilized uranium was readily solubilized by bicarbonate with concurrent release of phosphate into solution.

Photodegradation of uranium-citrate complex with uranium recovery

Upon exposure to visible light, uranyl citrate complex showed photodegradation of citric acid to acetic acid and carbon dioxide, with the precipitation of uranium as uranium trioxide (UO[sub 3][center dot]2H[sub 2]O). The initial pH and presence of oxygen affected the rate and extent of photochemical degradation of the complex, the formation of intermediate organic degradation products, and uranium speciation. Under aerobic conditions at pH 3.5, acetic, acetoacetic, 3-oxoglutaric, and malonic acids and acetone were detected; at pH 6.0, 3-oxoglutaric and acetic acids were present. The uranyl U(VI) ion was reduced to uranous U(IV) ion and was subsequently reoxidized to the hexavalent form and precipitated out of solution as uranium trioxide. Uranium trioxide precipitate was insoluble at near-neutral pH and was soluble in acidic pH (<4.1). Under anaerobic conditions, the uranyl citrate complex showed only partial (57%) degradation, and uranium was present in the reduced form as U(IV). Excess citric acid retarded the precipitation of uranium. 26 refs., 9 figs., 1 tab.

Denitrification in deep subsurface sediments

Abstract Dissimilatory nitrate reduction (denitrification) in subsurface sediments by indigenous microflora was investigated in samples obtained over a range of depths from 0 to 289 m. Denitrifying activity in sediment samples retrieved from similar stratigraphic horizons at four different sites was determined by measuring the accumulation of N2O using the acetylene blockage technique. Denitrification was detected in unamended samples which received only prereduced deionized water at almost all depths in all sediments sampled. The surface sediments showed the highest denitrification activity. In the deeper sediments, denitrifying activity was much higher in saturated sandy samples and lower or absent in drier clay samples. Addition of nitrate enhanced denitrification activity in all samples from below the water table down to the maximum depth sampled (289 m), while addition of a carbon (succinate) source in general had no stimulatory effect. These results show that denitrifying microorganisms were present...

Microbial transformations of radioactive wastes and environmental restoration through bioremediation

Abstract Bioremediation stabilizes and reclaims radionuclides and toxic metals from contaminated materials, soils, sediments, or wastes. The mechanism of microbial transformations of the radionuclides and toxic metals commonly found in energy wastes are summarized, and two processes for treating such wastes are described. In one process, anaerobic bacteria are used to concentrate, contain and stabilize the toxic metals and radionuclides in the waste, with a concurrent reduction in its volume. In the second process, the radionuclides and toxic metals are extracted from the wastes with citric acid which is then subjected to biodegradation, followed by photodegradation to recover the metals.

Toxicity of various anions associated with methoxyethyl methyl imidazolium-based ionic liquids on Clostridium sp.

We investigated the effects on the growth of the anaerobic bacterium, Clostridium sp., of the ionic liquid, 1-methoxyethyl-3-methyl imidazolium [MOEMIM](+), derived from imidazolium cation and paired with one of a variety of counter-ions, viz., tetrafluoroborate [BF₄]⁻, hexafluorophosphate [PF₆]⁻(,) trifluoroacetate [CF₃COO]⁻, bis(trifluoromethane)sulfonamide [Tf₂N]⁻, methane sulfonate [OMS], and 1-butyl-3-methyl imidazolium tetrafluoroborate [BMIM][BF₄]. These anions, in association with [MOEMIM](+) lowered the growth rate of the bacterium, showing the following trend: [Tf₂N]⁻ ≧ [PF₆]⁻ > [BF₄]⁻ > [CF₃COO]⁻ > [OMS]⁻. Anions incorporating fluorine were more toxic than those without it, and their toxicity rose with an increase in the number of fluorine atoms. Also, [MOEMIM](+)[BF₄]⁻ was less toxic than [BMIM](+)[BF₄]⁻, probably due to the presence of a methoxyethyl functional group integrated in the cation side chain.

Toxicity of imidazolium- and pyridinium-based ionic liquids and the co-metabolic degradation of N-ethylpyridinium tetrafluoroborate

We examined the effects of the ionic liquids (ILs), 1-butyl-3-methylimidazolium hexafluorophosphate [BMIM][PF₆], N-ethylpyridinium tetrafluoroborate [EtPy][BF₄], and N-ethylpyridinium trifluoroacetate [EtPy][CF₃COO] on Pseudomonas fluorescens, a ubiquitous soil bacterium. In the presence of 0.5- and 1% of [BMIM][PF₆] or [EtPy][CF₃COO] the growth of bacteria was inhibited, whereas exposing them to 1% [EtPy][BF₄] increased the lag period wherein bacteria adapt to growth conditions before continuing to grow. However, at higher concentrations (5% and 10%), no growth was observed. The inhibitory effects were evident by a decrease in the optical density of the culture, a decline in the consumption of the carbon source, citric acid, and a change in the size of the bacterium. At concentrations below 1%, [EtPy][BF₄] was metabolized by P. fluorescens in the presence of citric acid. Oxidation of the side alkyl-chain of [EtPy][BF₄] caused the accumulation of N-hydroxylethylpyridinium and pyridinium as major degradation products.

Biodegradation of pyridinium-based ionic liquids by an axenic culture of soil Corynebacteria

We investigated the biodegradation of ionic liquids N-ethylpyridinium tetrafluoroborate [EtPy]+[BF4]−, N-ethylpyridinium trifluoroacetate [EtPy]+[CF3COO]−, and 1-butyl-3-methylimidazolium hexafluorophosphate [BMIM]+[PF6]− by a soil bacterium isolated by an enrichment-culture technique. The bacterium identified as Corynebacterium sp. degraded the N-ethylpyridinium cation in the first two compounds when present as its sole carbon and nitrogen source without any obvious effects of the anion; however, [BMIM]+[PF6]− was not metabolized. We observed cleavage of the pyridinium ring and identified the resulting metabolites by ESI/MS/MS. We propose a degradation pathway.

Alkyl-methylimidazolium ionic liquids affect the growth and fermentative metabolism of Clostridium sp.

In this study, the effect of ionic liquids, 1-ethyl-3-methylimidazolium acetate [EMIM][Ac], 1-ethyl-3-methylimidazolium diethylphosphate [EMIM][DEP], and 1-methyl-3-methylimidazolium dimethylphosphate [MMIM][DMP] on the growth and glucose fermentation of Clostridium sp. was investigated. Among the three ionic liquids tested, [MMIM][DMP] was found to be least toxic. Growth of Clostridium sp. was not inhibited up to 2.5, 4 and 4 g L(-1) of [EMIM][Ac], [EMIM][DEP] and [MMIM][DMP], respectively. [EMIM][Ac] at <2.5 g L(-1), showed hormetic effect and stimulated the growth and fermentation by modulating medium pH. Total organic acid production increased in the presence of 2.5 and 2 g L(-1) of [EMIM][Ac] and [MMIM][DMP]. Ionic liquids had no significant influence on alcohol production at <2.5 g L(-1). Total gas production was affected by ILs at ≥ 2.5 g L(-1) and varied with type of methylimidazolium IL. Overall, the results show that the growth and fermentative metabolism of Clostridium sp. is not impacted by ILs at concentrations below 2.5 g L(-1).