Jonathan O. Sharp

N-Nitrosodimethylamine (NDMA) as a Drinking Water Contaminant: A Review

N-Nitrosodimethylamine (NDMA) is a member of a family of extremely potent carcinogens, the N-nitrosamines. Until recently, concerns about NDMA mainly focused on the presence of NDMA in food, consumer products, and polluted air. However, current concern focuses on NDMA as a drinking water contaminant resulting from reactions occurring during chlorination or via direct industrial contamination. Because of the relatively high concentrations of NDMA formed during wastewater chlorination, the intentional and unintentional reuse of municipal wastewater is a particularly important area of concern. Although ultraviolet (UV) treatment can effectively remove NDMA, there is considerable interest in the development of less expensive alternative treatment technologies. These alternative technologies include approaches for removing organic nitrogen-containing NDMA precursors prior to chlorination and the use of sunlight photolysis, and in situ bioremediation to remove NDMA and its precursors.

Non-uraninite Products of Microbial U(VI) Reduction

A promising remediation approach to mitigate subsurface uranium contamination is the stimulation of indigenous bacteria to reduce mobile U(VI) to sparingly soluble U(IV). The product of microbial uranium reduction is often reported as the mineral uraninite. Here, we show that the end products of uranium reduction by several environmentally relevant bacteria (Gram-positive and Gram-negative) and their spores include a variety of U(IV) species other than uraninite. U(IV) products were prepared in chemically variable media and characterized using transmission electron microscopy (TEM) and X-ray absorption spectroscopy (XAS) to elucidate the factors favoring/inhibiting uraninite formation and to constrain molecular structure/composition of the non-uraninite reduction products. Molecular complexes of U(IV) were found to be bound to biomass, most likely through P-containing ligands. Minor U(IV)-orthophosphates such as ningyoite [CaU(PO(4))(2)], U(2)O(PO(4))(2), and U(2)(PO(4))(P(3)O(10)) were observed in addition to uraninite. Although factors controlling the predominance of these species are complex, the presence of various solutes was found to generally inhibit uraninite formation. These results suggest a new paradigm for U(IV) in the subsurface, i.e., that non-uraninite U(IV) products may be found more commonly than anticipated. These findings are relevant for bioremediation strategies and underscore the need for characterizing the stability of non-uraninite U(IV) species in natural settings.

Bark beetle infestation impacts on nutrient cycling, water quality and interdependent hydrological effects

Bark beetle populations have drastically increased in magnitude over the last several decades leading to the largest-scale tree mortality ever recorded from an insect infestation on multiple wooded continents. When the trees die, the loss of canopy and changes in water and nutrient uptake lead to observable changes in hydrology and biogeochemical cycling. This review aims to synthesize the current research on the effects of the bark beetle epidemic on nutrient cycling and water quality while integrating recent and relevant hydrological findings, along with suggesting necessary future research avenues. Studies generally agree that snow depth will increase in infested forests, though the magnitude is uncertain. Changes in evapotranspiration are more variable as decreased transpiration from tree death may be offset by increased understory evapotranspiration and ground evaporation. As a result of such competing hydrologic processes that can affect watershed biogeochemistry along with the inherent variability of natural watershed characteristics, water quality changes related to beetle infestation are difficult to predict and may be regionally distinct. However, tree-scale changes to soil–water chemistry (N, P, DOC and base cation concentrations and composition) are being observed in association with beetle outbreaks which ultimately could lead to larger-scale responses. The different temporal and spatial patterns of bark beetle infestations due to different beetle and tree species lead to inconsistent infestation impacts. Climatic variations and large-scale watershed responses provide a further challenge for predictions due to spatial heterogeneities within a single watershed; conflicting reports from different regions suggest that hydrologic and water quality impacts of the beetle on watersheds cannot be generalized. Research regarding the subsurface water and chemical flow-paths and residence times after a bark beetle epidemic is lacking and needs to be rigorously addressed to best predict watershed or regional-scale changes to soil–water, groundwater, and stream water chemistry.

An Inducible Propane Monooxygenase Is Responsible for N-Nitrosodimethylamine Degradation by Rhodococcus sp. Strain RHA1

ABSTRACT Rhodococci are common soil heterotrophs that possess diverse functional enzymatic activities with economic and ecological significance. In this study, the correlation between gene expression and biological removal of the water contaminant N-nitrosodimethylamine (NDMA) is explored. NDMA is a hydrophilic, potent carcinogen that has gained recent notoriety due to its environmental persistence and emergence as a widespread micropollutant in the subsurface environment. In this study, we demonstrate that Rhodococcus sp. strain RHA1 can constitutively degrade NDMA and that activity toward this compound is enhanced by approximately 500-fold after growth on propane. Transcriptomic analysis of RHA1 and reverse transcriptase quantitative PCR assays demonstrate that growth on propane elicits the upregulation of gene clusters associated with (i) the oxidation of propane and (ii) the oxidation of substituted benzenes. Deletion mutagenesis of prmA, the gene encoding the large hydroxylase component of propane monooxygenase, abolished both growth on propane and removal of NDMA. These results demonstrate that propane monooxygenase is responsible for NDMA degradation by RHA1 and explain the enhanced cometabolic degradation of NDMA in the presence of propane.

Dissolved Organic Carbon Influences Microbial Community Composition and Diversity in Managed Aquifer Recharge Systems

ABSTRACT This study explores microbial community structure in managed aquifer recharge (MAR) systems across both laboratory and field scales. Two field sites, the Taif River (Taif, Saudi Arabia) and South Platte River (Colorado), were selected as geographically distinct MAR systems. Samples derived from unsaturated riverbed, saturated-shallow-infiltration (depth, 1 to 2 cm), and intermediate-infiltration (depth, 10 to 50 cm) zones were collected. Complementary laboratory-scale sediment columns representing low (0.6 mg/liter) and moderate (5 mg/liter) dissolved organic carbon (DOC) concentrations were used to further query the influence of DOC and depth on microbial assemblages. Microbial density was positively correlated with the DOC concentration, while diversity was negatively correlated at both the laboratory and field scales. Microbial communities derived from analogous sampling zones in each river were not phylogenetically significantly different on phylum, class, genus, and species levels, as determined by 16S rRNA gene pyrosequencing, suggesting that geography and season exerted less sway than aqueous geochemical properties. When field-scale communities derived from the Taif and South Platte River sediments were grouped together, principal coordinate analysis revealed distinct clusters with regard to the three sample zones (unsaturated, shallow, and intermediate saturated) and, further, with respect to DOC concentration. An analogous trend as a function of depth and corresponding DOC loss was observed in column studies. Canonical correspondence analysis suggests that microbial classes Betaproteobacteria and Gammaproteobacteria are positively correlated with DOC concentration. Our combined analyses at both the laboratory and field scales suggest that DOC may exert a strong influence on microbial community composition and diversity in MAR saturated zones.

Functional characterization of propane‐enhanced N‐nitrosodimethylamine degradation by two actinomycetales

Propane‐induced cometabolic degradation of n‐nitrosodimethylamine (NDMA) by two propanotrophs is characterized through kinetic, gene presence, and expression studies. After growth on propane, resting cells of Rhodococcus sp. RR1 possessed a maximum transformation rate (vmax,n) of 44 ± 5 µg NDMA (mg protein)−1 h−1; the rate for Mycobacterium vaccae (austroafricanum) JOB‐5 was modestly lower with vmax,n of 28 ± 3 µg NDMA (mg protein)−1 h−1. Both strains were capable of degrading environmentally relevant, trace quantities of NDMA to below the experimental limit of detection, calculated as 20 ng NDMA L−1. However, a comparison of half saturation constants (Ks,n) and NDMA degradation in the presence of propane revealed pronounced differences between the strains. The Ks,n for strain RR1 was 36 ± 10 µg NDMA L−1 while the propane concentration needed to inhibit NDMA rates by 50% (Kinh) occurred at 7,700 µg propane L−1 (R2 = 0.9669). In contrast, strain JOB‐5 had a markedly lower affinity for NDMA verses propane with a calculated Ks,n of 2,200 ± 1,000 µg NDMA L−1 and Kinh of 120 µg propane L−1 (R2 = 0.9895). Genomic and transcriptional investigations indicated that the functional enzymes involved in NDMA degradation and propane metabolism are different for each strain. For Rhodococcus sp. RR1, a putative propane monooxygenase (PrMO) was identified and implicated in NDMA oxidation. In contrast, JOB‐5 was not found to possess a PrMO homologue and two functionally analogous alkane monoxygenases (AlkMOs) were not induced by growth on propane. Differences between the PrMO in this Rhodococcus and the unidentified enzyme(s) in the Mycobacterium may explain differences in NDMA degradation and inhibition kinetics between these strains. Biotechnol. Bioeng. 2010;107: 924–932.

Biotransformation of Trace Organic Contaminants in Open-Water Unit Process Treatment Wetlands

The bottoms of shallow, open-water wetland cells are rapidly colonized by a biomat consisting of an assemblage of photosynthetic and heterotrophic microorganisms. To assess the contribution of biotransformation in this biomat to the overall attenuation of trace organic contaminants, transformation rates of test compounds measured in microcosms were compared with attenuation rates measured in a pilot-scale system. The biomat in the pilot-scale system was composed of diatoms (Staurosira construens) and a bacterial community dominated by β- and γ-Proteobacteria. Biotransformation was the dominant removal mechanism in the pilot-scale system for atenolol, metoprolol, and trimethoprim, while sulfamethoxazole and propranolol were attenuated mainly via photolysis. In microcosm experiments, biotransformation rates increased for metoprolol and propranolol when algal photosynthesis was supported by irradiation with visible light. Biotransformation rates increased for trimethoprim and sulfamethoxazole in the dark, when microbial respiration depleted dissolved oxygen concentrations within the biomat. During summer, atenolol, metoprolol, and propranolol were rapidly attenuated in the pilot-scale system (t1/2 < 0.5 d), trimethoprim and sulfamethoxazole were transformed more slowly (t1/2 ≈ 1.5-2 d), and carbamazepine was recalcitrant. The combination of biotransformation and photolysis resulted in overall transformation rates that were 10 to 100 times faster than those previously measured in vegetated wetlands, allowing for over 90% attenuation of all compounds studied except carbamazepine within an area similar to that typical of existing full-scale vegetated treatment wetlands.

Nitrate removal in shallow, open-water treatment wetlands.

The diffuse biomat formed on the bottom of shallow, open-water unit process wetland cells contains suboxic zones that provide conditions conducive to NO3(-) removal via microbial denitrification, as well as anaerobic ammonium oxidation (anammox). To assess these processes, nitrogen cycling was evaluated over a 3-year period in a pilot-scale wetland cell receiving nitrified municipal wastewater effluent. NO3(-) removal varied seasonally, with approximately two-thirds of the NO3(-) entering the cell removed on an annual basis. Microcosm studies indicated that NO3(-) removal was mainly attributable to denitrification within the diffuse biomat (i.e., 80 ± 20%), with accretion of assimilated nitrogen accounting for less than 3% of the NO3(-) removed. The importance of denitrification to NO3(-) removal was supported by the presence of denitrifying genes (nirS and nirK) within the biomat. While modest when compared to the presence of denitrifying genes, a higher abundance of the anammox-specific gene hydrazine synthase (hzs) at the biomat bottom than at the biomat surface, the simultaneous presence of NH4(+) and NO3(-) within the biomat, and NH4(+) removal coupled to NO2(-) and NO3(-) removal in microcosm studies, suggested that anammox may have been responsible for some NO3(-) removal, following reduction of NO3(-) to NO2(-) within the biomat. The annual temperature-corrected areal first-order NO3(-) removal rate (k20 = 59.4 ± 6.2 m yr(-1)) was higher than values reported for more than 75% of vegetated wetlands that treated water in which NO3(-) was the primary nitrogen species (e.g., nitrified secondary wastewater effluent and agricultural runoff). The inclusion of open-water cells, originally designed for the removal of trace organic contaminants and pathogens, in unit-process wetlands may enhance NO3(-) removal as compared to existing vegetated wetland systems.

Thioarsenic Species Associated with Increased Arsenic Release during Biostimulated Subsurface Sulfate Reduction

Introduction of acetate into groundwater at the Rifle Integrated Field Research Challenge (Rifle, CO) has been used for biostimulation aimed at immobilizing uranium. While a promising approach for lowering groundwater-associated uranium, a concomitant increase in soluble arsenic was also observed at the site. An array of field data was analyzed to understand spatial and temporal trends in arsenic release and possible correlations to speciation, subsurface redox conditions, and biogeochemistry. Arsenic release (up to 9 μM) was strongest under sulfate reducing conditions in areas receiving the highest loadings of acetate. A mixture of thioarsenate species, primarily trithioarsenate and dithioarsenate, were found to dominate arsenic speciation (up to 80%) in wells with the highest arsenic releases; thioarsenates were absent or minor components in wells with low arsenic release. Laboratory batch incubations revealed a strong preference for the formation of multiple thioarsenic species in the presence of the reduced precursors arsenite and sulfide. Although total soluble arsenic increased during field biostimulation, the termination of sulfate reduction was accompanied by recovery of soluble arsenic to concentrations at or below prestimulation levels. Thioarsenic species can be responsible for the transient mobility of sediment-associated arsenic during sulfidogenesis and should be considered when remediation strategies are implemented in sulfate-bearing, contaminated aquifers.

Enhanced biofilm production by a toluene-degrading Rhodococcus observed after exposure to perfluoroalkyl acids.

This study focuses on interactions between aerobic soil-derived hydrocarbon degrading bacteria and a suite of perfluorocarboxylic acids and perfluoroalkylsulfonates that are found in aqueous film-forming foams used for fire suppression. No effect on toluene degradation rate or induction time was observed when active cells of Rhodococcus jostii strain RHA1 were exposed to toluene and a mixture of perfluoroalkyl acids (PFAAs) including perfluorooctanoic acid (PFOA) and perfluorooctanesulfonate (PFOS) at concentrations near the upper bounds of groundwater relevance (11 PFAAs at 10 mg/L each). However, exposure to aqueous PFAA concentrations above 2 mg/L (each) was associated with enhanced aggregation of bacterial cells and significant increases in extracellular polymeric substance production. Flocculation was only observed during exponential growth and not elicited when PFAAs were added to resting incubations; analogous flocculation was also observed in soil enrichments. Aggregation was accompanied by 2- to 3-fold upregulation of stress-associated genes, sigF3 and prmA, during growth of this Rhodococcus in the presence of PFAAs. These results suggest that biological responses, such as microbial stress and biofilm formation, could be more prominent than suppression of co-contaminant biodegradation in subsurface locations where poly- and perfluoroalkyl substances occur with hydrocarbon fuels.