Geology topics

Mary A. Voytek

Publications and source records attributed to Mary A. Voytek.

At least 19 recordsLinked to original sources

USGS2016

The effects of co-contaminants and native wetland sediments on the activity and dominant transformation mechanisms of a 1,1,2,2-tetrachloroethane (TeCA)-degrading enrichment culture

Bioremediation strategies, including bioaugmentation with chlorinated ethene-degrading enrichment cultures, have been successfully applied in the cleanup of subsurface environments contaminated with tetrachloroethene (PCE) and/or trichloroethene (TCE). However, these compounds are frequently found in the environment as components of mixtures that may also contain chlorinated ethanes and methanes. Under these conditions, the implementation of bioremediation may be complicated by inhibition effects, particularly when multiple dehalorespirers are present. We investigated the ability of the 1,1,2,2-tetrachloroethane (TeCA)-dechlorinating culture WBC-2 to biotransform TeCA alone, or a mixture of TeCA plus PCE and carbon tetrachloride (CT), in microcosms. The microcosms contained electron donors provided to biostimulate the added culture and sediment collected from a wetland where numerous “hotspots” of contamination with chlorinated solvent mixtures exist. The dominant TeCA biodegradation mechanism mediated by the WBC-2 culture in the microcosms was different in the presence of these wetland sediments than in the sediment-free enrichment culture or in previous WBC-2 bioaugmented microcosms and column tests conducted with wetland sediment collected at nearby sites. The co-contaminants and their daughter products also inhibited TeCA biodegradation by WBC-2. These results highlight the need to conduct biodegradability assays at new sites, particularly when multiple contaminants and dehalorespiring populations are present.

Chemosphere↗

USGS2015

Association of anatase (TiO2) and microbes: unusual fossilization effect or a potential biosignature?

We combined microbial paleontology and molecular biology methods to study the Eyreville B drill core from the 35.3-Ma-old Chesapeake Bay impact structure,Virginia, USA. The investigated sample is a pyrite vein collected from the 1353.81-1353.89 m depth interval, located within a section of biotite granite. The granite is a pre-impact rock that was disrupted by the impact event. A search for inorganic (mineral) biosignatures revealed the presence of micron-size rod morphologies of anatase (TiO2) embedded in chlorite coatings on pyrite grains. Neither the Acridine Orange microbial probe nor deoxyribonucleic acid (DNA) extraction followed by polymerase chain reaction (PCR) amplifi cation showed the presence of DNA or ribonucleic acid (RNA) at the location of anatase rods, implying the absence of viable cells in the investigated area. A Nile Red microbial probe revealed the presence of lipids in the rods. Because most of the lipids are resistant over geologic time spans, they are good biomarkers, and they are an indicator of biogenicity for these possibly 35-Ma-old microbial fossils. The mineral assemblage suggests that rod morphologies are associated with low-temperature (<100 °C) hydrothermal alteration that involved aqueous fl uids. The temporal constraints on the anatase fossils are still uncertain because pre-impact alteration of the granite and postimpact heating may have provided identical conditions for anatase precipitation and microbial preservation.

Geological Society of America Bulletin↗

USGS2014

Integration of stable carbon isotope, microbial community, dissolved hydrogen gas, and 2 H H 2 O tracer data to assess bioaugmentation for chlorinated ethene degradation in fractured rocks

An in situ bioaugmentation (BA) experiment was conducted to understand processes controlling microbial dechlorination of trichloroethene (TCE) in groundwater at the Naval Air Warfare Center (NAWC), West Trenton, NJ. In the BA experiment, an electron donor (emulsified vegetable oil and sodium lactate) and a chloro-respiring microbial consortium were injected into a well in fractured mudstone of Triassic age. Water enriched in 2 H was also injected as a tracer of the BA solution, to monitor advective transport processes. The changes in concentration and the δ 13 C of TCE, cis-dichloroethene (cis-DCE), and vinyl chloride (VC); the δ 2 H of water; changes in the abundance of the microbial communities; and the concentration of dissolved H2 gas compared to pre- test conditions, provided multiple lines of evidence that enhanced biodegradation occurred in the injection well and in two downgradient wells. For those wells where the biodegradation was stimulated intensively, the sum of the molar chlorinated ethene (CE) concentrations in post-BA water was higher than that of the sum of the pre-BA background molar CE concentrations. The concentration ratios of TCE/(cis-DCE + VC) indicated that the increase in molar CE concentration may result from additional TCE mobilized from the rock matrix in response to the oil injection or due to desorption/diffusion. The stable carbon isotope mass-balance calculations show that the weighted average 13 C isotope of the CEs was enriched for around a year compared to the background value in a two year monitoring period, an effective indication that dechlorination of VC was occurring. Insights gained from this study can be applied to efforts to use BA in other fractured rock systems. The study demonstrates that a BA approach can substantially enhance in situ bioremediation not only in fractures connected to the injection well, but also in the rock matrix around the well due to processes such as diffusion and desorption. Because the effect of the BA was intensive only in wells where an amendment was distributed during injection, it is necessary to adequately distribute the amendments throughout the fractured rock to achieve substantial bioremediation. The slowdown in BA effect after a year is due to some extend to the decrease abundant of appropriate microbes, but more likely the decreased concentration of electron donor.

New Jersey↗

USGS2013

Hyporheic zone denitrification: controls on effective reaction depth and contribution to whole-stream mass balance

Stream denitrification is thought to be enhanced by hyporheic transport but there is little direct evidence from the field. To demonstrate at a field site, we injected 15 NO 3 − , Br (conservative tracer), and SF 6 (gas exchange tracer) and compared measured whole-stream denitrification with in situ hyporheic denitrification in shallow and deeper flow paths of contrasting geomorphic units. Hyporheic denitrification accounted for between 1 and 200% of whole-stream denitrification. The reaction rate constant was positively related to hyporheic exchange rate (greater substrate delivery), concentrations of substrates DOC and nitrate, microbial denitrifier abundance (nirS), and measures of granular surface area and presence of anoxic microzones. The dimensionless product of the reaction rate constant and hyporheic residence time, λ hz τ hz define a Damköhler number, Da den-hz that was optimal in the subset of hyporheic flow paths where Da den-hz ≈ 1. Optimal conditions exclude inefficient deep pathways transport where substrates are used up and also exclude inefficient shallow pathways that require repeated hyporheic entries and exits to complete the reaction. The whole-stream reaction significance, R s (dimensionless), was quantified by multiplying Da den-hz by the proportion of stream discharge passing through the hyporheic zone. Together these two dimensionless metrics, one flow-path scale and the other reach-scale, quantify the whole-stream significance of hyporheic denitrification. One consequence is that the effective zone of significant denitrification often differs from the full depth of the hyporheic zone, which is one reason why whole-stream denitrification rates have not previously been explained based on total hyporheic-zone metrics such as hyporheic-zone size or residence time.

Indiana↗

USGS2013

The effect of coal bed dewatering and partial oxidation on biogenic methane potential

Coal formation dewatering at a site in the Powder River Basin was associated with enhanced potential for secondary biogenic methane determined by using a bioassay. We hypothesized that dewatering can stimulate microbial activity and increase the bioavailability of coal. We analyzed one dewatered and two water-saturated coals to examine possible ways in which dewatering influences coal bed natural gas biogenesis by looking at differences with respect to the native coal microbial community, coal-methane organic intermediates, and residual coal oxidation potential. Microbial biomass did not increase in response to dewatering. Small Subunit rRNA sequences retrieved from all coals sampled represented members from genera known to be aerobic, anaerobic and facultatively anaerobic. A Bray Curtis similarity analysis indicated that the microbial communities in water-saturated coals were more similar to each other than to the dewatered coal, suggesting an effect of dewatering. There was a higher incidence of long chain and volatile fatty acid intermediates in incubations of the dewatered coal compared to the water-saturated coals, and this could either be due to differences in microbial enzymatic activities or to chemical oxidation of the coal associated with O 2 exposure. Dilute H 2 O 2 treatment of two fractions of structural coal (kerogen and bitumen + kerogen) was used as a proxy for chemical oxidation by O 2 . The dewatered coal had a low residual oxidation potential compared to the water-saturated coals. Oxidation with 5% H 2 O 2 did increase the bioavailability of structural coal, and the increase in residual oxidation potential in the water saturated coals was approximately equivalent to the higher methanogenic potential measured in the dewatered coal. Evidence from this study supports the idea that coal bed dewatering could stimulate biogenic methanogenesis through partial oxidation of the structural organics in coal once anaerobic conditions are restored.

Montana;Wyoming↗

USGS2013

A biogeochemical and genetic survey of acetylene fermentation by environmental samples and bacterial isolates

Anoxic samples (sediment and groundwater) from 13 chemically diverse field sites were assayed for their ability to consume acetylene (C 2 H 2 ). Over incubation periods ranging from &tilde; 10 to 80 days, selected samples from 7 of the 13 tested sites displayed significant C2H2 removal. No significant formation of ethylene was noted in these incubations; therefore, C 2 H 2 consumption could be attributed to acetylene hydratase (AH) rather than nitrogenase activity. This putative AH (PAH) activity was observed in only 21% of the total of assayed samples, while amplification of AH genes from extracted DNA using degenerate primers derived from Pelobacter acetylenicus occurred in even fewer (9.8%) samples. Acetylene-fermenting bacteria were isolated as a pure culture from the sediments of a tidal mudflat in San Francisco Bay (SFB93) and as an enrichment culture from freshwater Searsville Lake (SV7). Comparison of 16S rDNA clone libraries revealed that SFB93 was closely related to P. carbolinicus, while SV7 consisted of several unrelated bacteria. AH gene was amplified from SFB93 but not SV7. The inability of the primers to generate amplicons in the SV7 enrichment, as well as from several of the environmental samples that displayed PAH activity, implied that either the primers were too highly constrained in their specificity or that there was a different type of AH gene in these environmental samples than occurs in P. acetylenicus. The significance of this work with regard to the search for life in the outer Solar System, where C 2 HL 2 is abundant, is discussed.

Geomicrobiology Journal↗

USGS2013

Molecular techniques to distinguish morphologically similar Hydrilla verticillata , Egeria densa , Elodea nuttallii , and Elodea canadensis

The four submerged aquatic species, hydrilla (Hydrilla verticillata [monoecious and dioecious]), Brazilian waterweed (Egeria densa), Canadian waterweed (Elodea canadensis), and western waterweed (Elodea nuttallii), are difficult to positively identify because of their morphological similarity to each other, resulting in possible misidentification. This limits our ability to understand their past and present distribution, which is important in aquatic plant management. We investigated a molecular technique to identify these species, which are problematic because of their invasive nature on multiple continents. Approximately 100 samples of these species, ranging in age from 40-yr-old herbarium samples to recently collected plants, were collected from regions across the United States. The distribution and range of the samples collected in this research were compared to those reported in the literature. We confirmed information on the current wide distribution of both hydrilla biotypes in the United States and discovered that hydrilla had actually invaded the waterways near Washington, DC 6 yr earlier than originally reported. In addition, we found evidence of the confusion, dating back to the 1980s, between Canadian waterweed and western waterweed in the mid-Atlantic region of the United States. Canadian waterweed was previously reported as common and western waterweed as rare; however, our samples indicate the opposite is true. This information indicates there is a need for investigators to anticipate the spread of hydrilla populations to northern U.S. waterways, where it will compete with existing plant species, including Canadian and western waterweeds. Our ability to confirm distribution and pace of spread of invasive and noninvasive species will improve with increased application of molecular techniques.

Journal of Aquatic Plant Management↗

USGS2012

Impact disruption and recovery of the deep subsurface biosphere

Although a large fraction of the world's biomass resides in the subsurface, there has been no study of the effects of catastrophic disturbance on the deep biosphere and the rate of its subsequent recovery. We carried out an investigation of the microbiology of a 1.76 km drill core obtained from the ~35 million-year-old Chesapeake Bay impact structure, USA, with robust contamination control. Microbial enumerations displayed a logarithmic downward decline, but the different gradient, when compared to previously studied sites, and the scatter of the data are consistent with a microbiota influenced by the geological disturbances caused by the impact. Microbial abundance is low in buried crater-fill, ocean-resurge, and avalanche deposits despite the presence of redox couples for growth. Coupled with the low hydraulic conductivity, the data suggest the microbial community has not yet recovered from the impact ~35 million years ago. Microbial enumerations, molecular analysis of microbial enrichment cultures, and geochemical analysis showed recolonization of a deep region of impact-fractured rock that was heated to above the upper temperature limit for life at the time of impact. These results show how, by fracturing subsurface rocks, impacts can extend the depth of the biosphere. This phenomenon would have provided deep refugia for life on the more heavily bombarded early Earth, and it shows that the deeply fractured regions of impact craters are promising targets to study the past and present habitability of Mars.

Astrobiology↗

USGS2011

Microbial community structure of hydrothermal deposits from geochemically different vent fields along the Mid-Atlantic Ridge

To evaluate the effects of local fluid geochemistry on microbial communities associated with active hydrothermal vent deposits, we examined the archaeal and bacterial communities of 12 samples collected from two very different vent fields: the basalt-hosted Lucky Strike (37°17'N, 32°16.3'W, depth 1600-1750m) and the ultramafic-hosted Rainbow (36°13'N, 33°54.1'W, depth 2270-2330m) vent fields along the Mid-Atlantic Ridge (MAR). Using multiplexed barcoded pyrosequencing of the variable region 4 (V4) of the 16S rRNA genes, we show statistically significant differences between the archaeal and bacterial communities associated with the different vent fields. Quantitative polymerase chain reaction (qPCR) assays of the functional gene diagnostic for methanogenesis ( mcr A), as well as geochemical modelling to predict pore fluid chemistries within the deposits, support the pyrosequencing observations. Collectively, these results show that the less reduced, hydrogen-poor fluids at Lucky Strike limit colonization by strict anaerobes such as methanogens, and allow for hyperthermophilic microaerophiles, like Aeropyrum . In contrast, the hydrogen-rich reducing vent fluids at the ultramafic-influenced Rainbow vent field support the prevalence of methanogens and other hydrogen-oxidizing thermophiles at this site. These results demonstrate that biogeographical patterns of hydrothermal vent microorganisms are shaped in part by large scale geological and geochemical processes.

Environmental Microbiology↗

USGS2010

A record of phosphorus dynamics in oligotrophic lake sediment

Historical phosphorus (P) dynamics were studied using sediment cores from three oligotrophic, acidic lakes in Maine, USA. Long-term oligotrophy of these lakes is consistent with high sediment aluminum (as Al(OH) 3 ) concentrations, as Al inhibits internal P loading, even under reducing conditions. The role of microbially-mediated reactions in controlling redox conditions was evaluated by estimating microbial biomass and relative abundance of specific functional groups. Sediments were fractionated using a sequential chemical extraction technique and all lakes met criteria for P retention based on threshold sediment concentrations of Al, Fe, and P fractions as determined by (Kopáček et al. ( 2005 ) Limnol Oceanogr 52: 1147–1155). Sediment NaOH-extractable molybdate-reactive P (rP) and non-reactive P (nrP) represent P associated with non-reducible phases, and organic matter-related P, respectively. Total P (TP) does not decrease with sediment depth, as is typical of eutrophic lake sediments; however, nrP/TP decreases and rP/TP increases for all three lakes, indicating nrP mineralization without any significant upward diffusion and release into the hypolimnion; i.e. diagenesis of P is conservative within the sediment. Two diagenetic models were developed based on nrP and rP concentrations as a function of sediment age. The first model assumes a first-order decay of nrP, the rate coefficient being a function of time, and represents irreversible nrP mineralization, where the produced PO 4 is permanently sequestered by the sediment. The second model assumes a first-order reversible transformation between nrP and rP, representing biotic mineralization of organic P followed by incorporation of inorganic P into microbial biomass. Both models reflect preservation of TP with no loss to overlying water. The rate coefficients give us insight into qualities of the sediment that have affected mineralization and sequestration of phosphorus throughout the 210 Pb-dateable history of each lake. Similar models could be constructed for other lakes to help reconstruct their trophic histories. Paleolimnological reconstruction of the sediment P record in oligotrophic lakes shows mineralization of nrP to rP, but unlike the case in eutrophic lake sediments, sediment TP is preserved in these sediments.

Maine↗

USGS2010

Organic intermediates in the anaerobic biodegradation of coal to methane under laboratory conditions

Organic intermediates in coal fluids produced by anaerobic biodegradation of geopolymers in coal play a key role in the production of methane in natural gas reservoirs. Laboratory biodegradation experiments on sub-bituminous coal from Texas, USA, were conducted using bioreactors to examine the organic intermediates relevant to methane production. Production of methane in the bioreactors was linked to acetate accumulation in bioreactor fluid. Long chain fatty acids, alkanes (C 19 –C 36 ) and various low molecular weight aromatics, including phenols, also accumulated in the bioreactor fluid and appear to be the primary intermediates in the biodegradation pathway from coal-derived geopolymers to acetate and methane.

Texas↗

USGS2010

Stimulation of methane generation from nonproductive coal by addition of nutrients or a microbial consortium

Biogenic formation of methane from coal is of great interest as an underexploited source of clean energy. The goal of some coal bed producers is to extend coal bed methane productivity and to utilize hydrocarbon wastes such as coal slurry to generate new methane. However, the process and factors controlling the process, and thus ways to stimulate it, are poorly understood. Subbituminous coal from a nonproductive well in south Texas was stimulated to produce methane in microcosms when the native population was supplemented with nutrients (biostimulation) or when nutrients and a consortium of bacteria and methanogens enriched from wetland sediment were added (bioaugmentation). The native population enriched by nutrient addition included Pseudomonas spp., Veillonellaceae , and Methanosarcina barkeri . The bioaugmented microcosm generated methane more rapidly and to a higher concentration than the biostimulated microcosm. Dissolved organics, including long-chain fatty acids, single-ring aromatics, and long-chain alkanes accumulated in the first 39 days of the bioaugmented microcosm and were then degraded, accompanied by generation of methane. The bioaugmented microcosm was dominated by Geobacter sp., and most of the methane generation was associated with growth of Methanosaeta concilii . The ability of the bioaugmentation culture to produce methane from coal intermediates was confirmed in incubations of culture with representative organic compounds. This study indicates that methane production could be stimulated at the nonproductive field site and that low microbial biomass may be limiting in situ methane generation. In addition, the microcosm study suggests that the pathway for generating methane from coal involves complex microbial partnerships.

Texas↗

USGS2009

Multi-scale measurements and modeling of denitrification in streams with varying flow and nitrate concentration in the upper Mississippi River basin, USA

Denitrification is an important net sink for NO 3 − in streams, but direct measurements are limited and in situ controlling factors are not well known. We measured denitrification at multiple scales over a range of flow conditions and NO 3 − concentrations in streams draining agricultural land in the upper Mississippi River basin. Comparisons of reach-scale measurements (in-stream mass transport and tracer tests) with local-scale in situ measurements (pore-water profiles, benthic chambers) and laboratory data (sediment core microcosms) gave evidence for heterogeneity in factors affecting benthic denitrification both temporally (e.g., seasonal variation in NO 3 − concentrations and loads, flood-related disruption and re-growth of benthic communities and organic deposits) and spatially (e.g., local stream morphology and sediment characteristics). When expressed as vertical denitrification flux per unit area of streambed ( U denit , in μmol N m −2 h −1 ), results of different methods for a given set of conditions commonly were in agreement within a factor of 2–3. At approximately constant temperature (~20 ± 4°C) and with minimal benthic disturbance, our aggregated data indicated an overall positive relation between U denit (~0–4,000 μmol N m −2 h −1 ) and stream NO 3 − concentration (~20–1,100 μmol L −1 ) representing seasonal variation from spring high flow (high NO 3 − ) to late summer low flow (low NO 3 − ). The temporal dependence of U denit on NO 3 − was less than first-order and could be described about equally well with power-law or saturation equations (e.g., for the unweighted dataset, U denit ≈26 * [NO 3 − ] 0.44 or U denit ≈640 * [NO 3 − ]/[180 + NO 3 − ]; for a partially weighted dataset, U denit ≈14 * [NO 3 − ] 0.54 or U denit ≈700 * [NO 3 − ]/[320 + NO 3 − ]). Similar parameters were derived from a recent spatial comparison of stream denitrification extending to lower NO 3 − concentrations (LINX2), and from the combined dataset from both studies over 3 orders of magnitude in NO 3 − concentration. Hypothetical models based on our results illustrate: (1) U denit was inversely related to denitrification rate constant ( k 1 denit , in day −1 ) and vertical transfer velocity ( v f,denit , in m day −1 ) at seasonal and possibly event time scales; (2) although k 1 denit was relatively large at low flow (low NO 3 − ), its impact on annual loads was relatively small because higher concentrations and loads at high flow were not fully compensated by increases in U denit ; and (3) although NO 3 − assimilation and denitrification were linked through production of organic reactants, rates of NO 3 − loss by these processes may have been partially decoupled by changes in flow and sediment transport. Whereas k 1 denit and v f,denit are linked implicitly with stream depth, NO 3 − concentration, and(or) NO 3 − load, estimates of U denit may be related more directly to field factors (including NO 3 − concentration) affecting denitrification rates in benthic sediments. Regional regressions and simulations of benthic denitrification in stream networks might be improved by including a non-linear relation between U denit and stream NO 3 − concentration and accounting for temporal variation.

Biogeochemistry↗

USGS2009

Pore-water chemistry from the ICDP-USGS coer hole in the Chesapeake Bay impact structure--Implications for paleohydrology, microbial habitat, and water resources

We investigated the groundwater system of the Chesapeake Bay impact structure by analyzing the pore-water chemistry in cores taken from a 1766-m-deep drill hole 10 km north of Cape Charles, Virginia. Pore water was extracted using high-speed centrifuges from over 100 cores sampled from a 1300 m section of the drill hole. The pore-water samples were analyzed for major cations and anions, stable isotopes of water and sulfate, dissolved and total carbon, and bioavailable iron. The results reveal a broad transition between fresh and saline water from 100 to 500 m depth in the post-impact sediment section, and an underlying syn-impact section that is almost entirely filled with brine. The presence of brine in the lowermost post-impact section and the trend in the dissolved chloride with depth suggest a transport process dominated by molecular diffusion and slow, compaction-driven, upward flow. Major ion results indicate residual effects of diagenesis from heating, and a pre-impact origin for the brine. High levels of dissolved organic carbon (6-95 mg/L) and the distribution of electron acceptors indicate an environment that may be favorable for microbial activity throughout the drilled section. The concentration and extent of the brine is much greater than had previously been observed, suggesting its occurrence may be common in the inner crater. However, groundwater flow conditions in the structure may reduce the salt-water-intrusion hazard associated with the brine.

Special Paper of the Geological Society of America↗

USGS2008

Bioassay for estimating the biogenic methane-generating potential of coal samples

Generation of secondary biogenic methane in coal beds is likely controlled by a combination of factors such as the bioavailability of coal carbon, the presence of a microbial community to convert coal carbon to methane, and an environment supporting microbial growth and methanogenesis. A set of treatments and controls was developed to bioassay the bioavailability of coal for conversion to methane under defined laboratory conditions. Treatments included adding a well-characterized consortium of bacteria and methanogens (enriched from modern wetland sediments) and providing conditions to support endemic microbial activity. The contribution of desorbed methane in the bioassays was determined in treatments with bromoethane sulfonic acid, an inhibitor of microbial methanogenesis. The bioassay compared 16 subbituminous coal samples collected from beds in Texas (TX), Wyoming (WY), and Alaska (AK), and two bituminous coal samples from Pennsylvania (PA). New biogenic methane was observed in several samples of subbituminous coal with the microbial consortium added, but endemic activity was less commonly observed. The highest methane generation [80 µmol methane/g coal (56 scf/ton or 1.75 cm 3 /g)] was from a south TX coal sample that was collected from a non-gas-producing well. Subbituminous coals from the Powder River Basin, WY and North Slope Borough, AK contained more sorbed (original) methane than the TX coal sample and generated 0–23 µmol/g (up to 16 scf/ton or 0.5 cm 3 /g) new biogenic methane in the bioassay. Standard indicators of thermal maturity such as burial depth, nitrogen content, and calorific value did not explain differences in biogenic methane among subbituminous coal samples. No original methane was observed in two bituminous samples from PA, nor was any new methane generated in bioassays of these samples. The bioassay offers a new tool for assessing the potential of coal for biogenic methane generation, and provides a platform for studying the mechanisms involved in this economically important activity.

International Journal of Coal Geology↗

USGS2008

Microbial consortia development and microcosm and column experiments for enhanced bioremediation of chlorinated volatile organic compounds, West Branch Canal Creek wetland area, Aberdeen Proving Ground, Maryland

Chlorinated solvents, including 1,1,2,2-tetrachloroethane, tetrachloroethene, trichloroethene, carbon tetrachloride, and chloroform, are reaching land surface in localized areas of focused ground-water discharge (seeps) in a wetland and tidal creek in the West Branch Canal Creek area, Aberdeen Proving Ground, Maryland. In cooperation with the U.S. Army Garrison, Aberdeen Proving Ground, Maryland, the U.S. Geological Survey is developing enhanced bioremediation methods that simulate the natural anaerobic degradation that occurs without intervention in non-seep areas of the wetland. A combination of natural attenuation and enhanced bioremediation could provide a remedy for the discharging ground-water plumes that would minimize disturbance to the sensitive wetland ecosystem. Biostimulation (addition of organic substrate or nutrients) and bioaugmentation (addition of microbial consortium), applied either by direct injection at depth in the wetland sediments or by construction of a permeable reactive mat at the seep surface, were tested as possible methods to enhance anaerobic degradation in the seep areas. For the first phase of developing enhanced bioremediation methods for the contaminant mixtures in the seeps, laboratory studies were conducted to develop a microbial consortium to degrade 1,1,2,2-tetrachloroethane and its chlorinated daughter products under anaerobic conditions, and to test biostimulation and bioaugmentation of wetland sediment and reactive mat matrices in microcosms. The individual components required for the direct injection and reactive mat methods were then combined in column experiments to test them under groundwater- flow rates and contaminant concentrations observed in the field. Results showed that both direct injection and the reactive mat are promising remediation methods, although the success of direct injection likely would depend on adequately distributing and maintaining organic substrate throughout the wetland sediment in the seep area. For bioaugmentation, two mixed anaerobic cultures, named the 'West Branch Consortia' (WBC-1 and WBC-2), were developed by enrichment of wetland sediment collected from two contaminated sites in the study area where rapid and complete reductive dechlorination naturally occurs. WBC are capable of degrading 1,1,2,2-tetrachloroethane, 1,1,2-trichloroethane, 1,2-dichloroethane, tetrachloroethene, trichloroethene, cis- and trans-1,2-dichloroethene, and vinyl chloride to the non-chlorinated end-products ethene and ethane. In addition, the column experiments showed that the consortia could completely degrade carbon tetrachloride and chloroform, although they were not grown on these contaminants. No other cultures are known that can degrade the broad mixture of chlorinated alkanes, alkenes, and methanes as shown for WBC. WBC-2 (suspended in the culture media) is capable of complete dechlorination of 50 micromolar 1,1,2,2-tetrachloroethane to ethene in 1 to 2 days with little transient accumulation of chlorinated daughter products. Only about 5 percent of the clones sequenced from WBC-1 and WBC-2 were related to dechlorinating bacteria that have been studied previously in culture, indicating the presence of unknown dechlorinators. Dehalococcoides spp. comprised about 1 percent of WBC-1 and WBC-2, which is minor compared to the population size of about 30 percent in other dechlorinating consortia for chlorinated alkenes. Although both WBC-1 and WBC-2 showed efficient degradation in laboratory tests in this study, long-term cultivation of WBC-1, which was developed using hydrogen as the organic substrate, was determined to be infeasible. Thus, WBC-2, cultivated with lactate as the organic substrate, would be used in future tests. Nutrient (ammonia and phosphate mixture) addition to anaerobic microcosms constructed with wetland sediment and ground water collected from the study area showed some enhancement in the degradation rate of 1,1,2,2-tetrachloroethane, but degrada

Maryland↗

USGS2006

Characterization of a microbial consortium capable of rapid and simultaneous dechlorination of 1,1,2,2-tetrachloroethane and chlorinated ethane and ethene intermediates:

Mixed cultures capable of dechlorinating chlorinated ethanes and ethenes were enriched from contaminated wetland sediment at Aberdeen Proving Ground (APG) Maryland. The “West Branch Consortium” (WBC-2) was capable of degrading 1,1,2,2-tetrachloroethane (TeCA), trichloroethene (TCE), cis and trans 1,2-dichloroethene (DCE), 1,1,2-trichloroethane (TCA), 1,2-dichloroethane, and vinyl chloride to nonchlorinated end products ethene and ethane. WBC-2 dechlorinated TeCA, TCA, and cis DCE rapidly and simultaneously. A Clostridium sp. phylogenetically closely related to an uncultured member of a TCE-degrading consortium was numerically dominant in the WBC-2 clone library after 11 months of enrichment in culture. Clostridiales, including Acetobacteria, comprised 65% of the bacterial clones in WBC-2, with Bacteroides (14%), and epsilon Proteobacteria (14%) also numerically important. Methanogens identified in the consortium were members of the class Methanomicrobia, which includes acetoclastic methanogens. Dehalococcoides did not become dominant in the culture, although it was present at about 1% in the microbial population. The WBC-2 consortium provides opportunities for the in situ bioremediation of sites contaminated with mixtures of chlorinated ethenes and ethanes.

Bioremediation Journal↗

USGS2006

Methods for measuring denitrification: Diverse approaches to a difficult problem

Denitrification, the reduction of the nitrogen (N) oxides, nitrate (NO 3 − ) and nitrite (NO 2 − ), to the gases nitric oxide (NO), nitrous oxide (N 2 O), and dinitrogen (N 2 ), is important to primary production, water quality, and the chemistry and physics of the atmosphere at ecosystem, landscape, regional, and global scales. Unfortunately, this process is very difficult to measure, and existing methods are problematic for different reasons in different places at different times. In this paper, we review the major approaches that have been taken to measure denitrification in terrestrial and aquatic environments and discuss the strengths, weaknesses, and future prospects for the different methods. Methodological approaches covered include (1) acetylene-based methods, (2) 15 N tracers, (3) direct N 2 quantification, (4) N 2 :Ar ratio quantification, (5) mass balance approaches, (6) stoichiometric approaches, (7) methods based on stable isotopes, (8) in situ gradients with atmospheric environmental tracers, and (9) molecular approaches. Our review makes it clear that the prospects for improved quantification of denitrification vary greatly in different environments and at different scales. While current methodology allows for the production of accurate estimates of denitrification at scales relevant to water and air quality and ecosystem fertility questions in some systems (e.g., aquatic sediments, well-defined aquifers), methodology for other systems, especially upland terrestrial areas, still needs development. Comparison of mass balance and stoichiometric approaches that constrain estimates of denitrification at large scales with point measurements (made using multiple methods), in multiple systems, is likely to propel more improvement in denitrification methods over the next few years.

Ecological Applications↗