Determining the Reliability of Measuring Mercury Cycling Gene Abundance with Correlations with Mercury and Methylmercury Concentrations.

Methylmercury (MeHg) is a bioaccumulative toxic contaminant in many ecosystems, but factors governing its production are poorly understood. Recent work has shown that the anaerobic microbial conversion of mercury (Hg) to MeHg requires the Hg-methylation genes hgcAB and that these genes can be used as biomarkers in PCR-based estimators of Hg-methylator abundance. In an effort to determine reliable methods for assessing hgcA abundance and diversity and linking them to MeHg concentrations, multiple approaches were compared including metagenomic shotgun sequencing, 16S rRNA gene pyrosequencing and cloning/sequencing hgcAB gene products. Hg-methylator abundance was also determined by quantitative hgcA qPCR amplification and metaproteomics for comparison to the above measurements. Samples from eight sites were examined covering a range of total Hg (HgT; 0.03-14 mg kg-1 dry wt. soil) and MeHg (0.05-27 µg kg-1 dry wt. soil) concentrations. In the metagenome and amplicon sequencing of hgcAB diversity, the Deltaproteobacteria were the dominant Hg-methylators while Firmicutes and methanogenic Archaea were typically ∼50% less abundant. This was consistent with metaproteomics estimates where the Deltaproteobacteria were steadily higher. The 16S rRNA gene pyrosequencing did not have sufficient resolution to identify hgcAB+ species. Metagenomic and hgcAB results were similar for Hg-methylator diversity and clade-specific qPCR-based approaches for hgcA are only appropriate when comparing the abundance of a particular clade across various samples. Weak correlations between Hg-methylating bacteria and soil Hg concentrations were observed for similar environmental samples, but overall total Hg and MeHg concentrations poorly correlated with Hg-cycling genes.

Methylmercury sorption onto engineered materials.

Four commercially available sorbents (BioChar (BC), ThiolSAMMS® (TS), SediMite (SM), and Organoclay™ PM-199 (OC-199)) were tested for their ability to sorb methylmercury (MeHg) and MeHg complexed with dissolved organic matter (DOM). Testing sorption behavior with DOM is more representative of the environmental conditions and mercury speciation expected during in-situ remediation efforts. Isotherms were fit using a robust, iterative re-weighting scheme. This fitting approach improves upon the traditionally used indirect sorption method by removing the dependence between aqueous and solid phase concentrations in isotherm fitting. Developed isotherms show that without DOM, BC, TS, and SM adsorbed similar amounts of MeHg while OC-199 sorbed substantially less MeHg. Below an equilibrium concentration of 5.6 ng L-1 BC was the best performing sorbent, between 5.6 and 20.9 ng L-1 SM sorbed the most MeHg, and above an equilibrium concentration of 20.9 ng L-1 TS outperformed the other sorbents. BC and OC-199 showed indication of MeHg sorption saturation over the tested concentration range of 3.5-680 ng L-1. With DOM, SM outperformed the other sorbents at equilibrium concentrations less than 0.98 ng L-1 and TS was the superior MeHg:DOM sorbent at higher concentrations. MeHg:DOM sorption was controlled by DOM-sorbent interactions. DOM decreased MeHg sorption onto BC and SM whereas TS exhibited similar sorption with and without DOM. OC-199 had slightly higher MeHg uptake with DOM. East Fork Poplar Creek (EFPC), an industrially Hg contaminated site, was used as a case study example to build a relationship between aqueous and fish MeHg concentrations and subsequently compare the cost of sorbent materials required to meet regulatory objectives. For this case study, SM provided the most cost-effective sorbent option for in-situ remediation efforts to reduce aqueous MeHg concentrations.

Carbon Amendments Alter Microbial Community Structure and Net Mercury Methylation Potential in Sediments.

Neurotoxic methylmercury (MeHg) is produced by anaerobic Bacteria and Archaea possessing the genes hgcAB, but it is unknown how organic substrate and electron acceptor availability impacts the distribution and abundance of these organisms. We evaluated the impact of organic substrate amendments on mercury (Hg) methylation rates, microbial community structure, and the distribution of hgcAB+ microbes with sediments. Sediment slurries were amended with short-chain fatty acids, alcohols, or a polysaccharide. Minimal increases in MeHg were observed following lactate, ethanol, and methanol amendments, while a significant decrease (∼70%) was observed with cellobiose incubations. Postincubation, microbial diversity was assessed via 16S rRNA amplicon sequencing. The presence of hgcAB+ organisms was assessed with a broad-range degenerate PCR primer set for both genes, while the presence of microbes in each of the three dominant clades of methylators (Deltaproteobacteria, Firmicutes, and methanogenic Archaea) was measured with clade-specific degenerate hgcA quantitative PCR (qPCR) primer sets. The predominant microorganisms in unamended sediments consisted of Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria Clade-specific qPCR identified hgcA+Deltaproteobacteria and Archaea in all sites but failed to detect hgcA+Firmicutes Cellobiose shifted the communities in all samples to ∼90% non-hgcAB-containing Firmicutes (mainly Bacillus spp. and Clostridium spp.). These results suggest that either expression of hgcAB is downregulated or, more likely given the lack of 16S rRNA gene presence after cellobiose incubation, Hg-methylating organisms are largely outcompeted by cellobiose degraders or degradation products of cellobiose. These results represent a step toward understanding and exploring simple methodologies for controlling MeHg production in the environment.IMPORTANCE Methylmercury (MeHg) is a neurotoxin produced by microorganisms that bioacummulates in the food web and poses a serious health risk to humans. Currently, the impact that organic substrate or electron acceptor availability has on the mercury (Hg)-methylating microorganisms is unclear. To study this, we set up microcosm experiments exposed to different organic substrates and electron acceptors and assayed for Hg methylation rates, for microbial community structure, and for distribution of Hg methylators. The sediment and groundwater was collected from East Fork Poplar Creek in Oak Ridge, TN. Amendment with cellobiose (a lignocellulosic degradation by-product) led to a drastic decrease in the Hg methylation rate compared to that in an unamended control, with an associated shift in the microbial community to mostly nonmethylating Firmicutes This, along with previous Hg-methylating microorganism identification methods, will be important for identifying strategies to control MeHg production and inform future remediation strategies.

Temporal Dynamics of In-Field Bioreactor Populations Reflect the Groundwater System and Respond Predictably to Perturbation.

Temporal variability complicates testing the influences of environmental variability on microbial community structure and thus function. An in-field bioreactor system was developed to assess oxic versus anoxic manipulations on in situ groundwater communities. Each sample was sequenced (16S SSU rRNA genes, average 10,000 reads), and biogeochemical parameters are monitored by quantifying 53 metals, 12 organic acids, 14 anions, and 3 sugars. Changes in dissolved oxygen (DO), pH, and other variables were similar across bioreactors. Sequencing revealed a complex community that fluctuated in-step with the groundwater community and responded to DO. This also directly influenced the pH, and so the biotic impacts of DO and pH shifts are correlated. A null model demonstrated that bioreactor communities were driven in part not only by experimental conditions but also by stochastic variability and did not accurately capture alterations in diversity during perturbations. We identified two groups of abundant OTUs important to this system; one was abundant in high DO and pH and contained heterotrophs and oxidizers of iron, nitrite, and ammonium, whereas the other was abundant in low DO with the capability to reduce nitrate. In-field bioreactors are a powerful tool for capturing natural microbial community responses to alterations in geochemical factors beyond the bulk phase.

Development and Validation of Broad-Range Qualitative and Clade-Specific Quantitative Molecular Probes for Assessing Mercury Methylation in the Environment.

Two genes, hgcA and hgcB, are essential for microbial mercury (Hg) methylation. Detection and estimation of their abundance, in conjunction with Hg concentration, bioavailability, and biogeochemistry, are critical in determining potential hot spots of methylmercury (MeHg) generation in at-risk environments. We developed broad-range degenerate PCR primers spanning known hgcAB genes to determine the presence of both genes in diverse environments. These primers were tested against an extensive set of pure cultures with published genomes, including 13 Deltaproteobacteria, nine Firmicutes, and nine methanogenic Archaea genomes. A distinct PCR product at the expected size was confirmed for all hgcAB(+) strains tested via Sanger sequencing. Additionally, we developed clade-specific degenerate quantitative PCR (qPCR) primers that targeted hgcA for each of the three dominant Hg-methylating clades. The clade-specific qPCR primers amplified hgcA from 64%, 88%, and 86% of tested pure cultures of Deltaproteobacteria, Firmicutes, and Archaea, respectively, and were highly specific for each clade. Amplification efficiencies and detection limits were quantified for each organism. Primer sensitivity varied among species based on sequence conservation. Finally, to begin to evaluate the utility of our primer sets in nature, we tested hgcA and hgcAB recovery from pure cultures spiked into sand and soil. These novel quantitative molecular tools designed in this study will allow for more accurate identification and quantification of the individual Hg-methylating groups of microorganisms in the environment. The resulting data will be essential in developing accurate and robust predictive models of Hg methylation potential, ideally integrating the geochemistry of Hg methylation to the microbiology and genetics of hgcAB IMPORTANCE: The neurotoxin methylmercury (MeHg) poses a serious risk to human health. MeHg production in nature is associated with anaerobic microorganisms. The recent discovery of the Hg-methylating gene pair, hgcA and hgcB, has allowed us to design and optimize molecular probes against these genes within the genomic DNA for microorganisms known to methylate Hg. The protocols designed in this study allow for both qualitative and quantitative assessments of pure-culture or environmental samples. With these protocols in hand, we can begin to study the distribution of Hg-methylating organisms in nature via a cultivation-independent strategy.

Periphyton Biofilms Influence Net Methylmercury Production in an Industrially Contaminated System.

Mercury (Hg) methylation and methylmercury (MMHg) demethylation activity of periphyton biofilms from the industrially contaminated East Fork Poplar Creek, Tennessee (EFPC) were measured during 2014-2016 using stable Hg isotopic rate assays. 201HgII and MM202Hg were added to intact periphyton samples in ambient streamwater and the formation of MM201Hg and loss of MM202Hg were monitored over time and used to calculate first-order rate potentials for methylation and demethylation. The influences of location, temperature/season, light exposure and biofilm structure on methylation and demethylation potentials were examined. Between-site differences in net methylation for samples collected from an upstream versus downstream location were driven by differences in the demethylation rate potential (kd). In contrast, the within-site temperature-dependent difference in net methylation was driven by changes in the methylation rate potential (km). Samples incubated in the dark had lower net methylation due to lower km values than those incubated in the light. Disrupting the biofilm structure decreased km and resulted in lower net methylation. Overall, the measured rates resulted in a net excess of MMHg generated which could account for 3.71-7.88 mg d-1 MMHg flux in EFPC and suggests intact, actively photosynthesizing periphyton biofilms harbor zones of MMHg production.

Role of morphological growth state and gene expression in Desulfovibrio africanus strain Walvis Bay mercury methylation.

The biogeochemical transformations of mercury are a complex process, with the production of methylmercury, a potent human neurotoxin, repeatedly demonstrated in sulfate- and Fe(III)-reducing as well as methanogenic bacteria. However, little is known regarding the morphology, genes, or proteins involved in methylmercury generation. Desulfovibrio africanus strain Walvis Bay is a Hg-methylating δ-proteobacterium with a sequenced genome and has unusual pleomorphic forms. In this study, a relationship between the pleomorphism and Hg methylation was investigated. Proportional increases in the sigmoidal (regular) cell form corresponded with increased net MeHg production but decreased when the pinched cocci (persister) form became the major morphotype. D. africanus microarrays indicated that the ferrous iron transport genes (feoAB), as well as ribosomal genes and several genes whose products are predicted to have metal binding domains (CxxC), were up-regulated during exposure to Hg in the exponential phase. Whereas no specific methylation pathways were identified, the finding that Hg may interfere with iron transport and the correlation of growth-phase-dependent morphology with MeHg production are notable. The identification of these relationships between differential gene expression, morphology, and the growth-phase dependence of Hg transformations suggests that actively growing cells are primarily responsible for methylation, and so areas with ample carbon and electron-acceptor concentrations may also generate a higher proportion of methylmercury than more oligotrophic environments. The observation of increased iron transporter expression also suggests that Hg methylation may interfere with iron biogeochemical cycles.

Ecosystem service benefits to water users from perennial biomass production.

Although many agree that a transition to renewable energy sources is needed to avoid the climate consequences of continued reliance on fossil sources, price is a barrier. For renewable energy sources, including bioenergy, penetrating energy markets depends on lowering prices to compete with the price of fossil sources, but the tools used in decision making, such as supply curves, exclude non-market benefits from ecosystem services. Here, we extend the economic concept of an economic supply curve to account for ecosystem services co-produced with perennial biomass. We developed three new types of supply curves to visualize the increased supply of biomass ('sustainable supply') with sufficient water-quality benefits to offset biomass production costs. Using these tools, we show that the value of water-quality improvements could significantly reduce the break-even price of perennial feedstocks if it were available to farmers. In the most optimistic case, nearly half of potential biomass supply in a large tributary of the Mississippi river basin carried water purification value exceeding the cost of biomass production. Furthermore, adding the value to swimmers and waders offset production cost for over 90% of potential supply. Simulated benefits were context specific. For example, total value for water drinkers peaked at an intermediate level of fertilizer application. Geographically, benefits were highest in the eastern portion of the river basin. This research shows where the sustainable supply is needed and can generate value; the next step is to match this supply with credit buyers. Efforts to internalize the values of ecosystem services into biomass prices could help to meet Biden administration targets to meet 100% of sustainable aviation fuels.

Genome sequence of the mercury-methylating and pleomorphic Desulfovibrio africanus Strain Walvis Bay.

Desulfovibrio africanus strain Walvis Bay is an anaerobic sulfate-reducing bacterium capable of producing methylmercury (MeHg), a potent human neurotoxin. The mechanism of methylation by this and other organisms is unknown. We present the 4.2-Mb genome sequence to provide further insight into microbial mercury methylation and sulfate-reducing bacteria.

Genome sequence of the mercury-methylating strain Desulfovibrio desulfuricans ND132.

Desulfovibrio desulfuricans strain ND132 is an anaerobic sulfate-reducing bacterium (SRB) capable of producing methylmercury (MeHg), a potent human neurotoxin. The mechanism of methylation by this and other organisms is unknown. We present the 3.8-Mb genome sequence to provide further insight into microbial mercury methylation.

Mercury and other heavy metals influence bacterial community structure in contaminated Tennessee streams.

High concentrations of uranium, inorganic mercury [Hg(II)], and methylmercury (MeHg) have been detected in streams located in the Department of Energy reservation in Oak Ridge, TN. To determine the potential effects of the surface water contamination on the microbial community composition, surface stream sediments were collected 7 times during the year, from 5 contaminated locations and 1 control stream. Fifty-nine samples were analyzed for bacterial community composition and geochemistry. Community characterization was based on GS 454 FLX pyrosequencing with 235 Mb of 16S rRNA gene sequence targeting the V4 region. Sorting and filtering of the raw reads resulted in 588,699 high-quality sequences with lengths of >200 bp. The bacterial community consisted of 23 phyla, including Proteobacteria (ranging from 22.9 to 58.5% per sample), Cyanobacteria (0.2 to 32.0%), Acidobacteria (1.6 to 30.6%), Verrucomicrobia (3.4 to 31.0%), and unclassified bacteria. Redundancy analysis indicated no significant differences in the bacterial community structure between midchannel and near-bank samples. Significant correlations were found between the bacterial community and seasonal as well as geochemical factors. Furthermore, several community members within the Proteobacteria group that includes sulfate-reducing bacteria and within the Verrucomicrobia group appeared to be associated positively with Hg and MeHg. This study is the first to indicate an influence of MeHg on the in situ microbial community and suggests possible roles of these bacteria in the Hg/MeHg cycle.

A limited microbial consortium is responsible for extended bioreduction of uranium in a contaminated aquifer.

Subsurface amendments of slow-release substrates (e.g., emulsified vegetable oil [EVO]) are thought to be a pragmatic alternative to using short-lived, labile substrates for sustained uranium bioimmobilization within contaminated groundwater systems. Spatial and temporal dynamics of subsurface microbial communities during EVO amendment are unknown and likely differ significantly from those of populations stimulated by soluble substrates, such as ethanol and acetate. In this study, a one-time EVO injection resulted in decreased groundwater U concentrations that remained below initial levels for approximately 4 months. Pyrosequencing and quantitative PCR of 16S rRNA from monitoring well samples revealed a rapid decline in groundwater bacterial community richness and diversity after EVO injection, concurrent with increased 16S rRNA copy levels, indicating the selection of a narrow group of taxa rather than a broad community stimulation. Members of the Firmicutes family Veillonellaceae dominated after injection and most likely catalyzed the initial oil decomposition. Sulfate-reducing bacteria from the genus Desulforegula, known for long-chain fatty acid oxidation to acetate, also dominated after EVO amendment. Acetate and H(2) production during EVO degradation appeared to stimulate NO(3)(-), Fe(III), U(VI), and SO(4)(2-) reduction by members of the Comamonadaceae, Geobacteriaceae, and Desulfobacterales. Methanogenic archaea flourished late to comprise over 25% of the total microbial community. Bacterial diversity rebounded after 9 months, although community compositions remained distinct from the preamendment conditions. These results demonstrated that a one-time EVO amendment served as an effective electron donor source for in situ U(VI) bioreduction and that subsurface EVO degradation and metal reduction were likely mediated by successive identifiable guilds of organisms.

Importance of data management in a long-term biological monitoring program.

The long-term Biological Monitoring and Abatement Program (BMAP) has always needed to collect and retain high-quality data on which to base its assessments of ecological status of streams and their recovery after remediation. Its formal quality assurance, data processing, and data management components all contribute to meeting this need. The Quality Assurance Program comprehensively addresses requirements from various institutions, funders, and regulators, and includes a data management component. Centralized data management began a few years into the program when an existing relational database was adapted and extended to handle biological data. The database's main data tables and several key reference tables are described. One of the most important related activities supporting long-term analyses was the establishing of standards for sampling site names, taxonomic identification, flagging, and other components. The implemented relational database supports the transmittal of data to the Oak Ridge Environmental Information System (OREIS) as the permanent repository. We also discuss some limitations to our implementation. Some types of program data were not easily accommodated in the central systems, and many possible data-sharing and integration options are not easily accessible to investigators. From our experience we offer data management advice to other biologically oriented long-term environmental sampling and analysis programs.

Long-term benthic macroinvertebrate community monitoring to assess pollution abatement effectiveness.

The benthic macroinvertebrate community of East Fork Poplar Creek (EFPC) in East Tennessee was monitored for 18 years to evaluate the effectiveness of a water pollution control program implemented at a major United States (U.S.) Department of Energy facility. Several actions were implemented to reduce and control releases of pollutants into the headwaters of the stream. Four of the most significant actions were implemented during different time periods, which allowed assessment of each action. Macroinvertebrate samples were collected annually in April from three locations in EFPC (EFK24, EFK23, and EFK14) and two nearby reference streams from 1986 through 2003. Significant improvements occurred in the macroinvertebrate community at the headwater sites (EFK24 and EFK23) after implementation of each action, while changes detected 9 km further downstream (EFK14) could not be clearly attributed to any of the actions. Because the stream was impacted at its origin, invertebrate recolonization was primarily limited to aerial immigration, thus, recovery has been slow. As recovery progressed, abundances of small pollution-tolerant taxa (e.g., Orthocladiinae chironomids) decreased and longer lived taxa colonized (e.g., hydropsychid caddisflies, riffle beetles, Baetis). While assessments lasting three to four years may be long enough to detect a response to new pollution controls at highly impacted locations, more time may be needed to understand the full effects. Studies on the effectiveness of pollution controls can be improved if impacted and reference sites are selected to maximize spatial and temporal trending, and if a multidisciplinary approach is used to broadly assess environmental responses (e.g., water quality trends, invertebrate and fish community assessments, toxicity testing, etc.).

Increased nitrogen use efficiency in crop production can provide economic and environmental benefits.

Potential economic and environmental benefits of increasing nitrogen-use efficiency (NUE) are widely recognized but scarcely quantified. This study quantifies the effects of increased NUE on 1) the national agricultural economy using a simulation model of US agriculture and 2) regional water quality effects using a biogeochemical model for the Arkansas-White-Red river basin. National economic effects are reported for NUE improvement scenarios of 10%, 20%, 50%, and 100%, whereas regional water quality effects are estimated for a 20% NUE improvement scenario in the Arkansas-White-Red river basin. Simulating a 20% increase in NUE in row crops is shown to reduce N requirements by 1.4 million tonnes y-1 and increase farmer net profits by 1.6% ($743 million) per year by 2026 over the baseline simulation for the same period. For each 10% increase in NUE, annual farm revenues for commodity crops increased over the baseline by approximately $350 million per year by 2026. Changes in crop prices and land-use relative to the baseline were less than 2%. This suggests a net benefit even though fertilizer cost savings can result in increased cultivation of land, i.e., 'Jevon's paradox'. Results from the biogeochemical model of the Arkansas-White-Red river basin suggest that a 20% increase in NUE corresponds to a 5.72% reduction in nitrate loadings to freshwaters, with higher reductions in agricultural watersheds. The value of these reductions was estimated as $43 ha-1, for a total of $15.3 to 136.7 million yr-1 in avoided water treatment costs. After estimating the social value of increased NUE, we conclude with a discussion of potential strategies to increase efficiency and the research needed to achieve this goal. These include perennialization of the agricultural landscape, genetic crop improvement, targeted fertilizer application, and manipulation of the plant-root microbiome.

Microbial community changes in response to ethanol or methanol amendments for U(VI) reduction.

Microbial community responses to ethanol, methanol, and methanol plus humics amendments in relationship to U(VI) bioreduction were studied in laboratory microcosm experiments using sediments and ground water from a uranium-contaminated site in Oak Ridge, TN. The type of carbon source added, the duration of incubation, and the sampling site influenced the bacterial community structure upon incubation. Analysis of 16S rRNA gene clone libraries indicated that (i) bacterial communities found in ethanol- and methanol-amended samples with U(VI) reduction were similar due to the presence of Deltaproteobacteria and Betaproteobacteria (members of the families Burkholderiaceae, Comamonadaceae, Oxalobacteraceae, and Rhodocyclaceae); (ii) methanol-amended samples without U(VI) reduction exhibited the lowest diversity and the bacterial community contained 69.2 to 92.8% of the family Methylophilaceae; and (iii) the addition of humics resulted in an increase of phylogenetic diversity of Betaproteobacteria (Rodoferax, Polaromonas, Janthinobacterium, Methylophilales, and unclassified) and Firmicutes (Desulfosporosinus and Clostridium).

Cropland carbon fluxes in the United States: increasing geospatial resolution of inventory-based carbon accounting.

Net annual soil carbon change, fossil fuel emissions from cropland production, and cropland net primary production were estimated and spatially distributed using land cover defined by NASA's moderate resolution imaging spectroradiometer (MODIS) and by the USDA National Agricultural Statistics Service (NASS) cropland data layer (CDL). Spatially resolved estimates of net ecosystem exchange (NEE) and net ecosystem carbon balance (NECB) were developed. The purpose of generating spatial estimates of carbon fluxes, and the primary objective of this research, was to develop a method of carbon accounting that is consistent from field to national scales. NEE represents net on-site vertical fluxes of carbon. NECB represents all on-site and off-site carbon fluxes associated with crop production. Estimates of cropland NEE using moderate resolution (approximately 1 km2) land cover data were generated for the conterminous United States and compared with higher resolution (30-m) estimates of NEE and with direct measurements of CO2 flux from croplands in Illinois and Nebraska, USA. Estimates of NEE using the CDL (30-m resolution) had a higher correlation with eddy covariance flux tower estimates compared with estimates of NEE using MODIS. Estimates of NECB are primarily driven by net soil carbon change, fossil fuel emissions associated with crop production, and CO2 emissions from the application of agricultural lime. NEE and NECB for U.S. croplands were -274 and 7 Tg C/yr for 2004, respectively. Use of moderate- to high-resolution satellite-based land cover data enables improved estimates of cropland carbon dynamics.

Characterization of archaeal community in contaminated and uncontaminated surface stream sediments.

Archaeal communities from mercury and uranium-contaminated freshwater stream sediments were characterized and compared to archaeal communities present in an uncontaminated stream located in the vicinity of Oak Ridge, TN, USA. The distribution of the Archaea was determined by pyrosequencing analysis of the V4 region of 16S rRNA amplified from 12 streambed surface sediments. Crenarchaeota comprised 76% of the 1,670 archaeal sequences and the remaining 24% were from Euryarchaeota. Phylogenetic analysis further classified the Crenarchaeota as a Freshwater Group, Miscellaneous Crenarchaeota group, Group I3, Rice Cluster VI and IV, Marine Group I and Marine Benthic Group B; and the Euryarchaeota into Methanomicrobiales, Methanosarcinales, Methanobacteriales, Rice Cluster III, Marine Benthic Group D, Deep Sea Hydrothermal Vent Euryarchaeota 1 and Eury 5. All groups were previously described. Both hydrogen- and acetate-dependent methanogens were found in all samples. Most of the groups (with 60% of the sequences) described in this study were not similar to any cultivated isolates, making it difficult to discern their function in the freshwater microbial community. A significant decrease in the number of sequences, as well as in the diversity of archaeal communities was found in the contaminated sites. The Marine Group I, including the ammonia oxidizer Nitrosopumilus maritimus, was the dominant group in both mercury and uranium/nitrate-contaminated sites. The uranium-contaminated site also contained a high concentration of nitrate, thus Marine Group I may play a role in nitrogen cycle.

Donor-dependent extent of uranium reduction for bioremediation of contaminated sediment microcosms.

Bioremediation of uranium was investigated in microcosm experiments containing contaminated sediments from Oak Ridge, Tennessee to explore the importance of electron donor selection for uranium reduction rate and extent. In these experiments, all of the electron donors, including ethanol, glucose, methanol, and methanol with added humic acids, stimulated the reduction and immobilization of aqueous uranium by the indigenous microbial community. Uranium loss from solution began after the completion of nitrate reduction but essentially concurrent with sulfate reduction. When electron donor concentrations were normalized for their equivalent electron donor potential yield, the rates of uranium reduction were nearly equivalent for all treatments (0.55-0.95 micromol L(-1) d(-1)). Uranium reduction with methanol proceeded after a 15-d longer lag time relative to that of ethanol or glucose. Significant differences were not found with the inclusion of humic acids. The extent of U reduction in sediment slurries measured by XANES at various time periods after the start of the experiment increased in the order of ethanol (5-7% reduced at 77 and 153 d), glucose (49% reduced at 53 d), and methanol (93% reduced at 90 d). The microbial diversity of ethanol- and methanol-amended microcosms in their late stage of U reduction was analyzed with 16S rRNA gene amplification. Members of the Geobacteraceae were found in all microcosms as well as other potential uranium-reducing organisms, such as Clostridium and Desulfosporosinus. The effectiveness of methanol relative to ethanol at reducing aqueous and sediment-hosted uranium suggests that bioremediation strategies that encourage fermentative poising of the subsurface to a lower redox potential may be more effective for long-term uranium immobilization as compared with selecting an electron donor that is efficiently metabolized by known uranium-reducing microorganisms.

Energy use and carbon dioxide emissions from cropland production in the United States, 1990-2004.

Changes in cropland production and management influence energy consumption and emissions of CO(2) from fossil-fuel combustion. A method was developed to calculate on-site and off-site energy and CO(2) emissions for cropping practices in the United States at the county scale. Energy consumption and emissions occur on-site from the operation of farm machinery and occur off-site from the manufacture and transport of cropland production inputs, such as fertilizers, pesticides, and agricultural lime. Estimates of fossil-fuel consumption and associated CO(2) emissions for cropping practices enable (i) the monitoring of energy and emissions with changes in land management and (ii) the calculation and balancing of regional and national carbon budgets. Results indicate on-site energy use and total energy use (i.e., the sum of on-site and off-site) on U.S. croplands in 2004 ranged from 1.6 to 7.9 GJ ha(-1) yr(-1) and from 5.5 to 20.5 GJ ha(-1) yr(-1), respectively. On-site and total CO(2) emissions in 2004 ranged from 23 to 176 kg C ha(-1) yr(-1) and from 91 to 365 kg C ha(-1) yr(-1), respectively. During the period of this analysis (1990-2004), national total energy consumption for crop production ranged from 1204 to 1297 PJ yr(-1) (Petajoule = 1 x 10(15) Joule) with associated total fossil CO(2) emissions ranging from 21.5 to 23.2 Tg C yr(-1) (Teragram = 1 x 10(12) gram). The annual proportion of on-site CO(2) to total CO(2) emissions changed depending on the diversity of crops planted. Adoption of reduced tillage practices in the United States from 1990 to 2004 resulted in a net fossil emissions reduction of 2.4 Tg C.