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The identification and characterization of genes involved in the microbial oxidation of arsenite will contribute to our understanding of factors controlling As cycling in natural systems. Towards this goal, we recently characterized the widespread occurrence of aerobic arsenite oxidase genes (aroA-like) from pure-culture bacterial isolates, soils, sediments and geothermal mats, but were unable to detect these genes in all geothermal systems where we have observed microbial arsenite oxidation. Consequently, the objectives of the current study were to measure arsenite-oxidation rates in geochemically diverse thermal habitats in Yellowstone National Park (YNP) ranging in pH from 2.6 to 8, and to identify corresponding 16S rRNA and aroA genotypes associated with these arsenite-oxidizing environments. Geochemical analyses, including measurement of arsenite-oxidation rates within geothermal outflow channels, were combined with 16S rRNA gene and aroA functional gene analysis using newly designed primers to capture previously undescribed aroA-like arsenite oxidase gene diversity. The majority of bacterial 16S rRNA gene sequences found in acidic (pH 2.6-3.6) Fe-oxyhydroxide microbial mats were closely related to Hydrogenobaculum spp. (members of the bacterial order Aquificales), while the predominant sequences from near-neutral (pH 6.2-8) springs were affiliated with other Aquificales including Sulfurihydrogenibium spp., Thermocrinis spp. and Hydrogenobacter spp., as well as members of the Deinococci, Thermodesulfobacteria and beta-Proteobacteria. Modified primers designed around previously characterized and newly identified aroA-like genes successfully amplified new lineages of aroA-like genes associated with members of the Aquificales across all geothermal systems examined. The expression of Aquificales aroA-like genes was also confirmed in situ, and the resultant cDNA sequences were consistent with aroA genotypes identified in the same environments. The aroA sequences identified in the current study expand the phylogenetic distribution of known Mo-pterin arsenite oxidase genes, and suggest the importance of three prominent genera of the order Aquificales in arsenite oxidation across geochemically distinct geothermal habitats ranging in pH from 2.6 to 8.

Microbial reduction of arsenate [As(V)] to arsenite [As(III)] and the subsequent effects on As mobilization in contaminated mine tailings were studied under transport conditions. Molecular analysis of bacterial populations and traditional isolation techniques were used in conjunction with column experiments designed to observe relationships among pH (limed vs unlimed treatments), redox potential (Pt electrode), and mobilization of As. Liming increased pH values from approximately 4 to 8, resulting in a 5-fold increase in total As eluted from sterile columns. Elution of As from limed columns was further enhanced by microbial activity. As(III) was the predominant As species eluted from oxic, nonsterile columns. Conversely, in sterile treatments, As(V) was the predominant valence state in column effluent. Denaturing gradient gel electrophoresis coupled with sequence and phylogenetic analysis of 16S rRNA gene segments revealed that liming of the mine tailings stimulated specific Caulobacter-, Sphingomonas-, and Rhizobium-like populations. Pure culture isolates of these bacteria demonstrated the ability to rapidly reduce As(V) in aerated serum bottles. An intracellular As detoxification pathway was implicated in the reduction of As(V) by these isolates. These results indicate that microbial reduction of As(V) in As-contaminated soils may occur under aerobic conditions over relatively short time scales resulting in enhanced As mobilization.

Arsenic concentrations in 850 km of the Madison and Upper Missouri Rivers exceed the Montana State human health standard (0.24 Mu M). In addition, ground water As concentrations in the northern portion of the Lower Madison River Valley are above the federal drinking water standard (0.67 Mu M) and correlate with high soluble As levels in overlying soils. The objectives of this study were to determine processes affecting As solubility in soils of the Madison and Upper Missouri River Basins and assess potential impacts of long-term irrigation with As-rich river water on As mobility. Sixteen irrigated (>20 yr of irrigation) and nonirrigated (never irrigated) soils were sampled in fourmajor regions adjacent to the Madison and Upper Missouri Rivers. There were no significant differences (Alpha = 0.05) in total or solubleAs levels between irrigated and nonirrigated soils within any of thefour regions. In addition, sorption coefficients (Kd values) in six paired soils selected for additional chemical characterization were not significantly different (Alpha = 0.05) between irrigated and nonirrigated soils. Ammonium oxalate extractable Fe and Mn concentrations were found to be positively correlated with Kd and Olsen P concentration was found to be negatively correlated with Kd based on a multiple linear regression (r2 = 0.92). Sequential extractions performed for a subset of soils showed that labile As concentrations were similar between irrigated and nonirrigated soils. Soils that had been irrigated for >100 yr had additional As sorption capacities of at least 3 mg kg -1 in saturated column studies following 30 yr of simulated irrigation. Based onthis suite of data, it was determined that irrigation history has not significantly affected As solubility or attenuation capacities in soils of the study area. [ABSTRACT FROM AUTHOR]