Actinide biocolloid formation in brine by halophilic bacteria

The authors examined the ability of a halophilic bacterium (WIPP 1A) isolated from the Waste Isolation Pilot Plant (WIPP) site to accumulate uranium in order to determine the potential for biocolloid facilitated actinide transport. The bacterial cell surface functional groups involved in the complexation of the actinide were determined by titration. Uranium, added as uranyl nitrate, was removed from solution at pH 5 by cells but at pH 7 and 9 very little uranium was removed due to its limited solubility. Although present as soluble species, uranyl citrate at pH 5, 7, and 9, and uranyl carbonate at pH 9 were not removed by the bacterium because they were not bioavailable due to their neutral or negative charge. Addition of uranyl EDTA to brine at pH 5, 7, and 9 resulted in the immediate precipitation of U. Transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDS) analysis revealed that uranium was not only associated with the cell surface but also accumulated intracellularly as uranium-enriched granules. Extended X-ray absorption fine structure (EXAFS) analysis of the bacterial cells indicated the bulk sample contained more than one uranium phase. Nevertheless these results show the potential for the formation of actinide bearing bacterial biocolloids that are strictly regulated by the speciation and bioavailability of the actinide

Compliance of the Waste Isolation Pilot Plant with 40 CFR 194.24(b)

This paper presents aspects of DOE`s demonstration of compliance with the EPA regulation of the Waste Isolation Pilot Plant (WIPP). The WIPP, a geologic repository for transuranic (TRU) waste, is located 2150 feet below the ground surface in a bedded salt formation about 20 miles east of Carlsbad, NM. Performance of the WIPP as a repository requires that releases to the accessible environment not exceed the limits of the regulation 40 CFR Part 191(1) either when the WIPP is undisturbed, or if there is intrusion into the repository by drilling. In 1996, the EPA promulgated 40 CFR Part 194(2): the implementing regulation for 40 CFR Part 191. The regulatory subsection addressed here, 40 CFR 194.24(b), directs the DOE to identify and analyze the components and characteristics of the TRU waste that can impact performance of the WIPP repository, and thereby possibly impact waste containment. DOE must also analyze those waste characteristics and components that will not affect repository performance

A historical review of Waste Isolation Pilot Plant backfill development

Backfills have been part of Sandia National Laboratories' [Sandia's] Waste Isolation Pilot Plant [WIPP] designs for over twenty years. Historically, backfill research at Sandia has depended heavily on the changing mission of the WIPP facility. Early testing considered heat producing, high level, wastes. Bentonite/sand/salt mixtures were evaluated and studies focused on developing materials that would retard brine ingress, sorb radionuclides, and withstand elevated temperatures. The present-day backfill consists of pure MgO [magnesium oxide] in a pelletized form and is directed at treating the relatively low contamination level, non-heat producing, wastes actually being disposed of in the WIPP. Its introduction was motivated by the need to scavenging CO{sub 2} [carbon dioxide] from decaying organic components in the waste. However, other benefits, such as a substantial desiccating capacity, are also being evaluated. The MgO backfill also fulfills a statutory requirement for assurance measures beyond those needed to demonstrate compliance with the US Environmental Protection Agency [EPA] regulatory release limits. However, even without a backfill, the WIPP repository design still operates within EPA regulatory release limits

Interaction of Plutonium with Bacteria in the Repository Environment

Microorganisms in the nuclear waste repository environment may interact with plutonium through (1) sorption, (2) intracellular accumulation, and (3) transformation speciation. These interactions may retard or enhance the mobility of Pu by precipitation reactions, biocolloid formation, or production of more soluble species. Current and planned radioactive waste repository environments, such as deep subsurface halite and granite formations, are considered extreme relative to life processes in the near-surface terrestrial environment. There is a paucity of information on the biotransformation of radionuclides by microorganisms present in such extreme environments. In order to gain a better understanding of the interaction of plutonium with microorganisms present in the waste repository sites we investigated a pure culture (Halomonas sp.) and a mixed culture of bacteria (Haloarcula sinaiiensis, Marinobacter hydrocarbonoclasticus, Altermonas sp., and a {gamma}-proteobacterium) isolated from the Waste Isolation Pilot Plant (WIPP) site and an Acetobacterium sp. from alkaline groundwater at the Grimsel Test Site in Switzerland

Laboratory evaluation of colloidal actinide transport at the Waste Isolation Pilot Plant (WIPP): 1. crushed-dolomite column flow...

Colloid-facilitated transport of Pu, Am, U, Th, and Np has been recognized as a potentially important phenomenon affecting the performance of the Waste Isolation Pilot Plant (WIPP) facility being developed for safe disposal of transuranic radioactive waste. In a human intrusion scenario, actinide-bearing colloidal particles may be released from the repository and be transported by brines (approximately 0.8 to 3 molal ionic strength) through the Culebra, a thin fractured microcrystalline (mean grain size 2 micrometers) dolomite aquifer overlying the repository. Transport experiments were conducted using sieved, uniformly packed crushed Culebra rock or nonporous dolomite cleavage rhombohedra. Experiments with mineral fragments and fixed and live WIPP-relevant bacteria cultures showed significant levels of retardation due to physical filtration effects. Humic substances were not attenuated by the Culebra dolomite. Comparison of elution curves of latex microspheres in columns prepared with microcrystalline rock and nonporous rock showed minimal effect of Culebra micropores on colloid transport. These data form part of the basis to parameterize numerical codes being used to evaluate the performance of the WIPP

Actinide Biocolloid Formation in Brine by Halophilic Bacteria

The authors examined the ability of a halophilic bacterium (WIPP 1A) isolated from the Waste Isolation Pilot Plant (WIPP) site to accumulate uranium in order to determine the potential for biocolloid facilitated actinide transport. The bacterial cell surface functional groups involved in the complexation of the actinide were determined by titration. Uranium, added as uranyl nitrate, was removed from solution at pH 5 by cells but at pH 7 and 9 very little uranium was removed due to its limited solubility. Although present as soluble species, uranyl citrate at pH 5, 7, and 9, and uranyl carbonate at pH 9 were not removed by the bacterium because they were not bioavailable due to their neutral or negative charge. Addition of uranyl EDTA to brine at pH 5, 7, and 9 resulted in the immediate precipitation of U. Transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDS) analysis revealed that uranium was not only associated with the cell surface but also accumulated intracellularly as uranium-enriched granules. Extended X-ray absorption fine structure (EXAFS) analysis of the bacterial cells indicated the bulk sample contained more than one uranium phase. Nevertheless these results show the potential for the formation of actinide bearing bacterial biocolloids that are strictly regulated by the speciation and bioavailability of the actinide

Actinide Biocolloid Formation in Brine by Halophilic Bacteria

We examined the ability of a halophilic bacterium (WFP 1A) isolated from the Waste Isolation Pilot Plant (WIPP) site to accumulate uranium in order to determine the potential for biocolloid facilitated actinide transport. The bacterial cell Surface functional groups involved in the complexation of the actinide were determined by titration. Uranium, added as uranyl nitrate, was removed from solution at pH 5 by cells but at pH 7 and 9 very little uranium was removed due to its limited volubility. Although present as soluble species, uranyl citrate at pH 5, 7, and 9, and uranyl carbonate at pH 9 were not removed by the bacterium because they were not bioavailable due to their neutral or negative charge. Addition of uranyl EDTA to brine at pH 5, 7, and 9 resulted in the immediate precipitation of U. Transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDS) analysis revealed that uranium was not only associated with the cell surface but also accumulated intracellulary as uranium-enriched granules. Extended X-ray absorption fine structure (EXAFS) analysis, of the bacterial cells indicated the bulk sample contained more than one uranium phase. Nevertheless these results show the potential for the formation of actinide bearing bacterial biocolloids that are strictly regulated by the speciation and bioavailability of the actinide

Role of Microbes as Biocolloids in the Transport of Actinides from a Deep Underground Radioactive Waste Repository

We investigated the interaction of dissolved actinides Th, U, Np Zgpu, and Am, with a pure and a mixed culture of halophilic bactezia isolated from the Waste Isolation H.Iot Plant repository under anaerobic conditions to evaluate their potentiaI transport as biocolloids from the waste site. The sizes of the bacterial cells studied ranged from ().54 x 0.48 pm to 7.7 x 0.67pm Using sequential mimofiltration, we determined the ~~ation of actinides with fi-ee-living (mobile) bacterial cells suspended in a fluid medium containing. NaCl or M=W12 brine, at various phaes of their growth cycIes. The number of suspended kcteria rangy-d born 106 to 109 cells ml-*. Tine amount of actinide associatd with the wspend~ cell fraction (cakzdated & mol cell-*) was very Iow: Th, 10-*2; U, 10-1s - 10-lS; - ~ Np, 1o-15- 10-19; Pu, 10-ls -10-21 ; and h, 10-1* - 10-*9 ; and it varied with the bacteihl - CUIture studied. l%e differe&es in the asswiation are amibuted to the extent of bioamxmdation and biosorption by the bacteria pH, the compo&on of the brine, and the speziation and bioavaiIability of the actinides

Solid state NMR and X-ray studies of the structural evolution of nanocrystalline zirconia

A combination of X-ray techniques [diffraction and Zr K-edge absorption (EXAFS and XANES)] and multinuclear (H-1, C-13, O-17) solid-state NMR spectroscopy is employed to follow in detail the structural development of nanocrystalline zirconia. O-17 magic-angle spinning NMR spectroscopy of sol-gel produced undoped ZrO2 shows unequivocally that oxygen sites in the initial gel are monoclinic-like. This result is consistent with X-ray absorption measurements, which also suggest that the structures of the initial amorphous phases of doped and undoped samples produced by the hydroxide-precipitation and sol-gel methods are very similar. On crystallization, the local structure of the crystalline component is tetragonal, but a significant fraction of the sample remains disordered. Heating to higher temperatures results in conversion to monoclinic zirconia in undoped samples at room temperature. For sol-gel-produced ZrO2, C-13 NMR shows that loss of all of the organic fragments occurs prior to crystallization. The H-1 NMR experiments determined that the proton content remains significant until well above the crystallization temperature, so that the composition is not accurately described as ZrO2 until > 500 degreesC