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USGS2010

Laboratory chalcopyrite oxidation by Acidithiobacillus ferrooxidans: Oxygen and sulfur isotope fractionation

Laboratory experiments were conducted to simulate chalcopyrite oxidation under anaerobic and aerobic conditions in the absence or presence of the bacterium Acidithiobacillus ferrooxidans. Experiments were carried out with 3 different oxygen isotope values of water (?? 18 O H2O ) so that approach to equilibrium or steady-state isotope fractionation for different starting conditions could be evaluated. The contribution of dissolved O 2 and water-derived oxygen to dissolved sulfate formed by chalcopyrite oxidation was unambiguously resolved during the aerobic experiments. Aerobic oxidation of chalcopyrite showed 93 ?? 1% incorporation of water oxygen into the resulting sulfate during the biological experiments. Anaerobic experiments showed similar percentages of water oxygen incorporation into sulfate, but were more variable. The experiments also allowed determination of sulfate-water oxygen isotope fractionation, ?? 18 O SO4-H2O , of ~ 3.8??? for the anaerobic experiments. Aerobic oxidation produced apparent ?? SO4-H2O values (6.4???) higher than the anaerobic experiments, possibly due to additional incorporation of dissolved O 2 into sulfate. ?? 34 S SO4 values are ~ 4??? lower than the parent sulfide mineral during anaerobic oxidation of chalcopyrite, with no significant difference between abiotic and biological processes. For the aerobic experiments, a small depletion in ?? 34 S SO4 of ~- 1.5 ?? 0.2??? was observed for the biological experiments. Fewer solids precipitated during oxidation under aerobic conditions than under anaerobic conditions, which may account for the observed differences in sulfur isotope fractionation under these contrasting conditions. ?? 2009 Elsevier B.V.

Chemical Geology↗

USGS2020

Sequential biodegradation of 1,2,4-trichlorobenzene at oxic-anoxic groundwater interfaces in model laboratory columns

Halogenated organic solvents such as chlorobenzenes (CBs) are frequent groundwater contaminants due to legacy spills. When contaminated anaerobic groundwater discharges into surface water through wetlands and other transition zones, aeration can occur from various physical and biological processes at shallow depths, resulting in oxic-anoxic interfaces (OAIs). This study investigated the potential for 1,2,4-trichlorobenzene (1,2,4-TCB) biodegradation at OAIs. A novel upflow column system was developed to create stable anaerobic and aerobic zones, simulating a natural groundwater OAI. Two columns containing (1) sand and (2) a mixture of wetland sediment and sand were operated continuously for 295 days with varied doses of 0.14-1.4 mM sodium lactate (NaLac) as a model electron donor. Both column matrices supported anaerobic reductive dechlorination and aerobic degradation of 1,2,4-TCB spatially separated between anaerobic and aerobic zones. Reductive dechlorination produced a mixture of di- and monochlorobenzene daughter products, with estimated zero-order dechlorination rates up to 31.3 µM/hr. Aerobic CB degradation, limited by available dissolved oxygen, occurred for 1,2,4-TCB and all dechlorinated daughter products. Initial reductive dechlorination did not enhance the overall observed extent or rate of subsequent aerobic CB degradation. Increasing NaLac dose increased the extent of reductive dechlorination, but suppressed aerobic CB degradation at 1.4 mM NaLac due to increased oxygen demand. 16S-rRNA sequencing of biofilm microbial communities revealed strong stratification of functional anaerobic and aerobic organisms between redox zones including the sole putative reductive dechlorinator detected in the columns, Dehalobacter. The sediment mixture column supported enhanced reductive dechlorination compared to the sand column at all tested NaLac doses and growth of Dehalobacter populations up to 4.1×108 copies/g (51% relative abundance), highlighting the potential benefit of sediments in reductive dechlorination processes. Results from these model systems suggest both substantial anaerobic and aerobic CB degradation can co-occur along the OAI at contaminated sites where bioavailable electron donors and oxygen are both present.

Journal of Contaminant Hydrology↗

USGS2015

A pan-Arctic synthesis of CH 4 and CO 2 production from anoxic soil incubations

Permafrost thaw can alter the soil environment through changes in soil moisture, frequently resulting in soil saturation, a shift to anaerobic decomposition, and changes in the plant community. These changes, along with thawing of previously frozen organic material, can alter the form and magnitude of greenhouse gas production from permafrost ecosystems. We synthesized existing methane (CH 4 ) and carbon dioxide (CO 2 ) production measurements from anaerobic incubations of boreal and tundra soils from the geographic permafrost region to evaluate large-scale controls of anaerobic CO 2 and CH 4 production and compare the relative importance of landscape-level factors (e.g., vegetation type and landscape position), soil properties (e.g., pH, depth, and soil type), and soil environmental conditions (e.g., temperature and relative water table position). We found fivefold higher maximum CH 4 production per gram soil carbon from organic soils than mineral soils. Maximum CH 4 production from soils in the active layer (ground that thaws and refreezes annually) was nearly four times that of permafrost per gram soil carbon, and CH 4 production per gram soil carbon was two times greater from sites without permafrost than sites with permafrost. Maximum CH 4 and median anaerobic CO 2 production decreased with depth, while CO 2 :CH 4 production increased with depth. Maximum CH 4 production was highest in soils with herbaceous vegetation and soils that were either consistently or periodically inundated. This synthesis identifies the need to consider biome, landscape position, and vascular/moss vegetation types when modeling CH 4 production in permafrost ecosystems and suggests the need for longer-term anaerobic incubations to fully capture CH 4 dynamics. Our results demonstrate that as climate warms in arctic and boreal regions, rates of anaerobic CO 2 and CH 4 production will increase, not only as a result of increased temperature, but also from shifts in vegetation and increased ground saturation that will accompany permafrost thaw.

Global Change Biology↗

USGS2015

Hydrogeologic characterization and assessment of bioremediation of chlorinated benzenes and benzene in wetland areas, Standard Chlorine of Delaware, Inc. Superfund Site, New Castle County, Delaware, 2009-12

Wetlands at the Standard Chlorine of Delaware, Inc. Superfund Site (SCD) in New Castle County, Delaware, are affected by contamination with chlorobenzenes and benzene from past waste storage and disposal, spills, leaks, and contaminated groundwater discharge. In cooperation with the U.S. Environmental Protection Agency, the U.S. Geological Survey began an investigation in June 2009 to characterize the hydrogeology and geochemistry in the wetlands and assess the feasibility of monitored natural attenuation and enhanced bioremediation as remedial strategies. Groundwater flow in the wetland study area is predominantly vertically upward in the wetland sediments and the underlying aquifer, and groundwater discharge accounts for a minimum of 47 percent of the total discharge for the subwatershed of tidal Red Lion Creek. Thus, groundwater transport of contaminants to surface water could be significant. The major contaminants detected in groundwater in the wetland study area included benzene, monochlorobenzene, and tri- and di-chlorobenzenes. Shallow wetland groundwater in the northwest part of the wetland study area was characterized by high concentrations of total chlorinated benzenes and benzene (maximum about 75,000 micrograms per liter [μg/L]), low pH, and high chloride. In the northeast part of the wetland study area, wetland groundwater had low to moderate concentrations of total chlorinated benzenes and benzene (generally not greater than 10,000 μg/L), moderate pH, and high sulfate concentrations. Concentrations in the groundwater in excess of 1 percent of the solubility of the individual chlorinated benzenes indicate that a contaminant source is present in the wetland sediments as dense nonaqueous phase liquids (DNAPLs). Consistently higher contaminant concentrations in the shallow wetland groundwater than deeper in the wetland sediments or the aquifer also indicate a continued source in the wetland sediments, which could include dissolution of DNAPLs and desorption from the sediments. When highly reducing, methanogenic, or sulfate-reducing conditions existed in the wetland groundwater, molar composition of the volatile organic compounds (VOCs) showed that chlorobenzene and benzene were predominant, indicating biodegradation of the chlorinated benzenes through reductive dechlorination pathways. Temporal changes in redox conditions between 2009 and 2011–12 have shifted the locations in the wetland study area where reductive dechlorination is evident. Microbial community analyses of sediment showed relatively high cell numbers and diversity of populations ( Dehalococcoides , Dehalobacter , Desulfitobacterium , and Geobacter ) that are known to contain species capable of reductive dechlorination, confirming groundwater geochemistry evidence of the occurrence of reductive dechlorination. Natural attenuation was not sufficient, however, to reduce total VOC concentrations along upward groundwater flowpaths in the wetland sediments, most likely due to the additional source of contaminants in the upper sediments. In situ microcosms that were unamended except for the addition of 13 C-labeled contaminants in some treatments, confirmed that the native microbial community was able to biodegrade the higher chlorinated benzenes through reductive dechlorination and that 1,2-dichlorobenzene, chlorobenzene, and benzene could be degraded to carbon dioxide through oxidation pathways. Microcosms that were bioaugmented with the anaerobic dechlorinating consortium WBC-2 and deployed in the wetland sediments showed reductive dechlorination of tri-, di-, and monochlorobenzene, and 13 C-chlorobenzene treatments showed complete degradation of chlorobenzene to carbon dioxide under anaerobic conditions. Experiments with a continuous flow, fixed-film bioreactor seeded with native microorganisms in groundwater from the wetland area showed both aerobic and anaerobic biodegradation of dichlorobenzenes, monochlorobenzene, and benzene, although monochlorobenzene and benzene degradation rates decreased under anaerobic conditions compared to aerobic conditions. In two bioreactors with established biofilms of WBC-2, percent removals of all chlorinated benzene compounds (medians of 86 to 94 percent) under anaerobic conditions were as high as those observed for the bioreactors seeded only with native microorganisms from the site groundwater, and benzene removal was greater in the WBC-2 bioaugmented bioreactors. The high percent removals in the WBC-2 bioreactors without the need for an acclimation period indicates that the same dechlorinators are involved in the chlorinated benzene degradation as those for the chlorinated ethanes and ethenes that the culture was developed to degrade. The ability of the WBC-2 culture to completely reduce the chlorinated benzenes and benzene, even in the presence of high sulfate and sulfide concentrations, is unique for known dechlorinating cultures. The availability of the established culture WBC-2, as well as the ability of the native wetland microbial community to degrade the site contaminants under anaerobic and aerobic conditions, provides flexibility in considering bioremediation options for the wetland areas at SCD.

Delaware↗

USGS2007

Oxygen and sulfur isotope systematics of sulfate produced by bacterial and abiotic oxidation of pyrite

To better understand reaction pathways of pyrite oxidation and biogeochemical controls on ??18O and ??34S values of the generated sulfate in acid mine drainage (AMD) and other natural environments, we conducted a series of pyrite oxidation experiments in the laboratory. Our biological and abiotic experiments were conducted under aerobic conditions by using O2 as an oxidizing agent and under anaerobic conditions by using dissolved Fe(III)aq as an oxidant with varying ??18OH2O values in the presence and absence of Acidithiobacillus ferrooxidans. In addition, aerobic biological experiments were designed as short- and long-term experiments where the final pH was controlled at ???2.7 and 2.2, respectively. Due to the slower kinetics of abiotic sulfide oxidation, the aerobic abiotic experiments were only conducted as long term with a final pH of ???2.7. The ??34SSO4 values from both the biological and abiotic anaerobic experiments indicated a small but significant sulfur isotope fractionation (???-0.7???) in contrast to no significant fractionation observed from any of the aerobic experiments. Relative percentages of the incorporation of water-derived oxygen and dissolved oxygen (O2) to sulfate were estimated, in addition to the oxygen isotope fractionation between sulfate and water, and dissolved oxygen. As expected, during the biological and abiotic anaerobic experiments all of the sulfate oxygen was derived from water. The percentage incorporation of water-derived oxygen into sulfate during the oxidation experiments by O2 varied with longer incubation and lower pH, but not due to the presence or absence of bacteria. These percentages were estimated as 85%, 92% and 87% from the short-term biological, long-term biological and abiotic control experiments, respectively. An oxygen isotope fractionation effect between sulfate and water (??18 OSO4 s(-) H2 O) of ???3.5??? was determined for the anaerobic (biological and abiotic) experiments. This measured ??18 OSO42 - s(-) H2 O value was then used to estimate the oxygen isotope fractionation effects (??18 OSO42 - s(-) O2) between sulfate and dissolved oxygen in the aerobic experiments which were -10.0???, -10.8???, and -9.8??? for the short-term biological, long-term biological and abiotic control experiments, respectively. Based on the similarity between ??18OSO4 values in the biological and abiotic experiments, it is suggested that ??18OSO4 values cannot be used to distinguish biological and abiotic mechanisms of pyrite oxidation. The results presented here suggest that Fe(III)aq is the primary oxidant for pyrite at pH < 3, even in the presence of dissolved oxygen, and that the main oxygen source of sulfate is water-oxygen under both aerobic and anaerobic conditions. ?? 2007 Elsevier Ltd. All rights reserved.

Geochimica et Cosmochimica Acta↗

USGS2003

Inverse modeling of BTEX dissolution and biodegradation at the Bemidji, MN crude-oil spill site

The U.S. Geological Survey (USGS) solute transport and biodegradation code BIOMOC was used in conjunction with the USGS universal inverse modeling code UCODE to quantify field-scale hydrocarbon dissolution and biodegradation at the USGS Toxic Substances Hydrology Program crude-oil spill research site located near Bemidji, MN. This inverse modeling effort used the extensive historical data compiled at the Bemidji site from 1986 to 1997 and incorporated a multicomponent transport and biodegradation model. Inverse modeling was successful when coupled transport and degradation processes were incorporated into the model and a single dissolution rate coefficient was used for all BTEX components. Assuming a stationary oil body, we simulated benzene, toluene, ethylbenzene, m , p -xylene, and o -xylene (BTEX) concentrations in the oil and ground water, respectively, as well as dissolved oxygen. Dissolution from the oil phase and aerobic and anaerobic degradation processes were represented. The parameters estimated were the recharge rate, hydraulic conductivity, dissolution rate coefficient, individual first-order BTEX anaerobic degradation rates, and transverse dispersivity. Results were similar for simulations obtained using several alternative conceptual models of the hydrologic system and biodegradation processes. The dissolved BTEX concentration data were not sufficient to discriminate between these conceptual models. The calibrated simulations reproduced the general large-scale evolution of the plume, but did not reproduce the observed small-scale spatial and temporal variability in concentrations. The estimated anaerobic biodegradation rates for toluene and o -xylene were greater than the dissolution rate coefficient. However, the estimated anaerobic biodegradation rates for benzene, ethylbenzene, and m , p -xylene were less than the dissolution rate coefficient. The calibrated model was used to determine the BTEX mass balance in the oil body and groundwater plume. Dissolution from the oil body was greatest for compounds with large effective solubilities (benzene) and with large degradation rates (toluene and o -xylene). Anaerobic degradation removed 77% of the BTEX that dissolved into the water phase and aerobic degradation removed 17%. Although goodness-of-fit measures for the alternative conceptual models were not significantly different, predictions made with the models were quite variable.

Minnesota↗

USGS1988

Microbial transformations of azaarenes in creosite-contaminated soil and ground water: Laboratory and field studies

Azaarenes or aromatic nitrogen heterocycles are a class of compounds found in wood-preservative wastes containing creosote. The fate and movement of these compounds in contaminated aquifers is not well understood. Water-quality studies in an aquifer contaminated with creosote near Pensacola, Florida, indicated that ground water was contaminated with several azaarenes and their oxygenated and alkylated derivatives, suggesting that these oxygenated compounds may be products of microbial transformation reactions. Accordingly, laboratory studies were designed to investigate the fate of these compounds. Under aerobic conditions, soil pseudomonads isolated from creosote-contaminated soil converted quinoline to 2(1H)quinoline that subsequently was degraded to unknown products. A methanogenic consortium isolated from an anaerobic sewage digestor, in presence of ground-water and creosote-contaminated soil, converted quinoline, isoquinoline, and 4-methylquinoline to their respective oxygenated analogs. In addition, N-, C-, and O-methylated analogs of oxygenated azaarenes were identified by gas chromatography-mass spectrometry (GC-MS) in aerobic cultures. Under the experimental conditions, 2-methylquinoline was biorefractory. Presence of similar biotransformation products in anaerobic cultures and contaminated ground water from the Pensacola site provided further evidence that these compounds indeed were mivrobial transformation products. Stable isotope labeling studies indicated that the source of the oxygen atom for this hydroxylation reaction under aerobic and anaerobic conditions was water. A mechanism was proposed for this hydroxylation reaction. Whereas parent azaarenes are biodegradable in both anaerobic and aerobic zones, oxygenated and alkylated analogs are more biorefractory and, hence, persistent in anaerobic zones of contaminated aquifers.

Water Science and Technology↗

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↗

USGS2026

Treatability study to evaluate bioremediation of trichloroethene at Site K, former Twin Cities Army Ammunition Plant, Arden Hills, Minnesota, 2020–22

Executive Summary Chlorinated solvents, including trichloroethene (TCE) and other chlorinated volatile organic compounds (cVOCs), are widespread contaminants that can be treated by bioremediation approaches that enhance anaerobic reductive dechlorination. Reductive dechlorination can be enhanced either through the addition of an electron donor (biostimulation) or the addition of a known dechlorinating culture (bioaugmentation) along with an electron donor. Although bioremediation has been applied at many TCE-contaminated groundwater sites, application in source zones at sites where residual dense nonaqueous phase liquid (DNAPL) is present is more limited. In this study, laboratory and field treatability tests were completed to evaluate the potential application of anaerobic bioremediation for a shallow groundwater plume containing TCE in a perched alluvial aquifer at Site K, former Twin Cities Army Ammunition Plant, Arden Hills, Minnesota, which was on the National Priorities List as the New Brighton/Arden Hills Superfund site until 2019. In addition to the presence of residual DNAPL at the site, temporal variability in groundwater flow directions and input of oxygenated recharge were possible complicating factors for the application of enhanced anaerobic biodegradation in the shallow plume. The Site K plume extends beneath the footprint of Building 103, which was demolished in 2006, and soil excavations to a maximum depth of 6 feet (ft) below ground surface in 2014 were known to leave some deeper contaminated soil in place in the TCE source area. Groundwater treatment at the site, formalized as part of the 1997 Record of Decision, has been in operation since 1986 and consists of an extraction trench at the downgradient edge of the plume to collect groundwater, which is then pumped to an on-site air stripper. Groundwater concentrations in the plume have been relatively stable since treatment began, indicating a continued source of TCE in the aquifer. The desire for a destructive remedy that would enhance the removal of cVOCs in the aquifer at Site K and shorten the remediation timeframe led the U.S. Army to request that the U.S. Geological Survey conduct a groundwater treatability study to assess bioremediation. This report describes the U.S. Geological Survey bioremediation treatability study conducted during 2020–22, including pre-design site characterization to assist in formulating the bioremediation approach, laboratory experiments to support the design of the field pilot test, and implementation and 1-year performance monitoring results for the pilot test. Pre-design site characterization included the collection of soil cores for cVOC analysis and lithologic descriptions and the re-installment of three wells to obtain hydrologic measurements and initial groundwater chemistry. Relatively flat head gradients were measured at the site, and substantial decreases in water-level elevations occurred from spring to summer (May–July 2021). Continuous water-level monitoring indicated a rapid response to precipitation. Groundwater flow velocities were consistently less than 0.5 foot per day, and the pilot bioremediation test was therefore designed with short lateral distances (about 5 ft) between injection and individual monitoring points. Soil analyses confirmed that high volatile organic compound contamination was left in place in the source area. The highest concentrations were near or in clay at the base of the perched aquifer. Concentrations of cVOCs measured in the replaced wells were consistent with historical data and had a maximum TCE concentration of 57,700 micrograms per liter (μg/L), indicative of nearby residual DNAPL based on the general rule of observed concentrations exceeding 1 percent of solubility. The primary TCE daughter product detected was 1,2-cis-dichloroethene (cisDCE), which indicated limited reductive dechlorination in the plume. Groundwater in both the source and downgradient areas was relatively reducing during the pre-design characterization, particularly in the source area where methane concentrations greater than 400 μg/L were measured. Initial laboratory tests conducted using native aquifer microorganisms from the three replacement wells showed that anaerobic TCE biodegradation rates were low when biostimulated with the addition of sodium lactate as an electron donor, also known as a carbon donor, and resulted in the production of only cisDCE. Addition of a known dechlorinating culture, WBC-2, however, resulted in rapid biodegradation and production of ethene, verifying complete reductive dechlorination of TCE. Microcosms constructed with aquifer soil collected from the site were used to evaluate other electron donors besides lactate to support reductive dechlorination by WBC-2, including corn syrup as an alternative fast-release compound and whey, soy-based vegetable oil, and 3-D Microemulsion (Regenesis, San Clemente, California) as slow-release compounds. First-order rate constants for total organic chlorine removal in these WBC-2 amended microcosms were greatest with either lactate or vegetable oil as the donor, ranging between 0.061 and 0.047 per day or corresponding half-lives of 11–15 days. Testing of commercial products in other WBC-2-bioaugmented microcosms led to selection for the field pilot test of an emulsified vegetable oil product that also contained some sodium lactate as a fast-release donor. Delaying the addition of WBC-2 relative to the donor in the microcosms resulted in the most rapid overall biodegradation rates. The selected design for the pilot test utilized three separate test plots, each about 30-ft wide and 60-ft long: plots GS1 and GS2 in the source area of the plume and plot GS3 in the downgradient area of the plume near the excavation trench. Each test plot had one injection well, one monitoring well upgradient from the injection point, and 12 surrounding monitoring wells in a grid to capture variable groundwater flow directions. Donor injections, which included a bromide tracer, were completed in October 2021, immediately following baseline sampling, and the WBC-2 culture was injected about 40 days later, between November 30 and December 2, 2021. Performance monitoring conducted until December 2022 included hydrologic measurements and analyses of cVOCs, redox-sensitive constituents, dissolved organic carbon, bromide, volatile fatty acids, compound-specific carbon isotopes, and microbial communities. The biogeochemical data collected during the pilot tests in the three treatment plots showed that enhanced, complete reductive dechlorination of cVOCs in the groundwater was achieved in the GS1 and GS3 plots. In contrast, evidence of distribution of the injected amendments and subsequent biodegradation was limited in GS2, which was in an area of more heterogeneous soil lithology and low water table elevations. The molar composition of volatile organic compounds in the GS1 and GS3 plots was dominated by ethene in wells that were reached by the injected amendments by the end of the monitoring period. In the GS1 and GS3 plots, similar patterns were observed of cVOC concentrations decreasing to near detection levels, or below, at some wells sampled in July and October 2022, whereas ethene became dominant and indicated sustained complete reductive dechlorination. Baseline cVOC concentrations were more than a factor of 10 higher in the groundwater in the GS1 plot than in GS3, but no apparent inhibition of complete dechlorination occurred. As expected from the initial pre-design site data and the laboratory experiments, enhanced dissolution of residual DNAPL coupled to biodegradation was evident in the GS1 plot, where a marked increase in dichloroethene (DCE) above the initial baseline and upgradient TCE and DCE concentrations occurred. DCE concentrations subsequently declined where DNAPL dissolution was evident, concurrent with production of vinyl chloride and then predominantly ethene. Thus, overall biodegradation rates outpaced the DNAPL dissolution and desorption and DCE production in the source area. This success in complete degradation to predominantly ethene was achieved even in areas where the DCE concentrations reached a maximum of about 30,000 μg/L. Compound specific isotope analysis of carbon in TCE, cisDCE, trans-1,2-dichloroethene, and vinyl chloride was conducted to provide another line of evidence of the occurrence and extent of anaerobic biodegradation. Along a flow path in each plot that was affected by the injected amendments, carbon isotopes in the TCE and daughter cVOCs in the groundwater became isotopically heavier, indicating biodegradation. Enhanced biodegradation rates calculated from the field tests in GS1 and GS3 showed half-lives of 36.9–75.3 days for DCE degradation and 9.48–38.5 days for ethene production. Notably, these ethene production rates calculated from the field tests are consistent with the results of WBC-2-bioaugmented microcosms amended with either lactate or vegetable oil, which had half-lives for total organic chlorine removal that ranged from 11 to 15 days. These rates indicated rapid enhanced biodegradation, which is promising for application of a full-scale bioremediation remedy. Ultimately, however, the mass of residual or sorbed TCE in the aquifer that remains accessible for dissolution and biodegradation would likely control the time required for a full-scale bioremediation effort to achieve performance goals for TCE and cisDCE specified in the Record of Decision for Site K. The field pilot tests showed that the relatively low hydraulic head gradients and temporal changes in groundwater flow directions in the shallow aquifer would add complexity to a full-scale bioremediation effort. The radius of influence (ROI) at GS1 and GS3 (16.3 ft and 12.7 ft, respectively) were close to the design ROI of 15 ft. The estimated ROI at GS2 was about four times the design ROI, but may be less reliable at this location owing to groundwater flow direction. In addition, the low temperatures following WBC-2 injection in late November to early December 2021, in combination with the low hydraulic head gradients, were probably major factors in the delay observed before the onset of enhanced biodegradation following injection of the culture. Additional test injections could be beneficial to optimize the timing of donor and culture injections with the variable temperatures and hydraulic head in the shallow aquifer.

Minnesota↗

USGS1998

Microbial mineralization of VC and DCE under different terminal electron accepting conditions

Production of 14CO2 from [1,2-14C] dichloroethene (DCE) or [1,2-14C] vinyl chloride (VC) was quantified in aquifer and stream-bed sediment microcosms to evaluate the potential for microbial mineralization as a pathway for DCE and VC biodegradation under aerobic, Fe(III)-reducing, SO4-reducing, and methanogenic conditions. Mineralization of [1,2-14C] DCE and [1,2-14C] VC to 14CO2 decreased under increasingly reducing conditions, but significant mineralization was observed for both sediments even under anaerobic conditions. VC mineralization decreased in the order of aerobic > Fe(III)-reducing > SO4-reducing > methanogenic conditions. For both sediments, VC mineralization was greater than DCE mineralization under all electron-accepting conditions examined. For both sediments, DCE mineralization was at least two times greater under aerobic conditions than under anaerobic conditions. Although significant microbial mineralization of DCE was observed under anaerobic conditions, recovery of 14CO2 did not differ substantially between anaerobic treatments.

Anaerobe↗

USGS2013

Residence time control on hot moments of net nitrate production and uptake in the hyporheic zone

The retention capacity for biologically available nitrogen within streams can be influenced by dynamic hyporheic zone exchange, a process that may act as either a net source or net sink of dissolved nitrogen. Over 5 weeks, nine vertical profiles of streambed chemistry (NO 3 - and NH 4 + ) were collected above two beaver dams along with continuous high-resolution vertical hyporheic flux data. The results indicate a non-linear relation of net NO 3 - production followed by net uptake in the hyporheic zone as a function of residence time. This Lagrangian-based relation is consistent through time and across varied morphology (bars, pools, glides) above the dams, even though biogeochemical and environmental factors varied. The empirical continuum between net NO 3 - production and uptake and residence time is useful for identifying two crucial residence time thresholds: the transition to anaerobic respiration, which corresponds to the time of peak net nitrate production, and the net sink threshold, which is defined by a net uptake in NO 3 - relative to streamwater. Short-term hyporheic residence time variability at specific locations creates hot moments of net production and uptake, enhancing NO 3 - production as residence times approach the anaerobic threshold, and changing zones of net NO 3 - production to uptake as residence times increase past the net sink threshold. The anaerobic and net sink thresholds for beaver-influenced streambed morphology occur at much shorter residence times (1.3 h and 2.3 h, respectively) compared to other documented hyporheic systems, and the net sink threshold compares favorably to the lower boundary of the anaerobic threshold determined for this system with the new oxygen Damkohler number. The consistency of the residence time threshold values of NO 3 - cycling in this study, despite environmental variability and disparate morphology, indicates that NO 3 - hot moment dynamics are primarily driven by changes in physical hydrology and associated residence times.

Hydrological Processes↗

USGS1998

Field and laboratory evidence for intrinsic biodegradation of vinyl chloride contamination in a Fe(III)-reducing aquifer

Intrinsic bioremediation of chlorinated ethenes in anaerobic aquifers previously has not been considered feasible, due, in large part, to 1) the production of vinyl chloride during microbial reductive dechlorination of higher chlorinated contaminants and 2) the apparent poor biodegradability of vinyl chloride under anaerobic conditions. In this study, a combination of field geochemical analyses and laboratory radiotracer ([1,2-14C] vinyl chloride) experiments was utilized to assess the potential for intrinsic biodegradation of vinyl chloride contamination in an Fe(III)-reducing, anaerobic aquifer. Microcosm experiments conducted under Fe(III)-reducing conditions with material from the Fe(III)-reducing, chlorinated-ethene contaminated aquifer demonstrated significant oxidation of [1,2-14C] vinyl chloride to 14CO2 with no detectable production of ethene or other reductive dehalogenation products. Rates of degradation derived from the microcosm experiments (0.9-1.3% d-1) were consistent with field-estimated rates (0.03-0.2% d-1) of apparent vinyl chloride degradation. Field estimates of apparent vinyl chloride biodegradation were calculated using two distinct approaches; 1) a solute dispersion model and 2) a mass balance assessment. These findings demonstrate that degradation under Fe(III) reducing conditions can be an environmentally significant mechanism for intrinsic bioremediation of vinyl chloride in anaerobic ground-water systems.

Journal of Contaminant Hydrology↗

USGS2001

Effect of redox conditions on MTBE biodegradation in surface water Sediments

Microbial degradation of methyl tert-butyl ether (MTBE) was observed in surface water-sediment microcosms under anaerobic conditions. The efficiency and products of anaerobic MTBE biodegradation were dependent on the predominant terminal electron-accepting conditions. In the presence of substantial methanogenic activity, MTBE biodegradation was nominal and involved reduction of MTBE to the toxic product, tert-butyl alcohol (TBA). In the absence of significant methanogenic activity, accumulation of [14C]TBA generally decreased, and mineralization of [U-14C]MTBE to 14CO2 generally increased as the oxidative potential of the predominant terminal electron acceptor increased in the order of SO4, Fe(III), Mn(IV) < NO3 < O2. Microbial mineralization of MTBE to CO2 under Mn(IV)or SO4-reducing conditions has not been reported previously. The results of this study indicate that microorganisms inhabiting the sediments of streams and lakes can degrade MTBE effectively under a range of anaerobic terminal electron-accepting conditions. Thus, anaerobic bed sediment microbial processes may provide a significant environmental sink for MTBE in surface water systems throughout the United States.

Environmental Science & Technology↗

USGS2008

Persistence of pathogenic prion protein during simulated wastewater treatment processes

Transmissible spongiform encephalopathies (TSEs, prion diseases) are a class of fatal neurodegenerative diseases affecting a variety of mammalian species including humans. A misfolded form of the prion protein (PrP TSE) is the major, if not sole, component of the infectious agent. Prions are highly resistant to degradation and to many disinfection procedures suggesting that, if prions enter wastewater treatment systems through sewers and/or septic systems (e.g., from slaughterhouses, necropsy laboratories, rural meat processors, private game dressing) or through leachate from landfills that have received TSE-contaminated material, prions could survive conventional wastewater treatment Here, we report the results of experiments examining the partitioning and persistence of PrPTSE during simulated wastewater treatment processes including activated and mesophilic anaerobic sludge digestion. Incubation with activated sludge did not result in significant PrPTSE degradation. PrPTSE and prion infectivity partitioned strongly to activated sludge solids and are expected to enter biosolids treatment processes. A large fraction of PrPTSE survived simulated mesophilic anaerobic sludge digestion. The small reduction in recoverable PrPTSE after 20-d anaerobic sludge digestion appeared attributable to a combination of declining extractability with time and microbial degradation. Our results suggest that if prions were to enter municipal wastewater treatment systems, most would partition to activated sludge solids, survive mesophilic anaerobic digestion, and be present in treated biosolids. ?? 2008 American Chemical Society.

Environmental Science & Technology↗

USGS2005

Microbial H2 cycling does not affect δ2H values of ground water

Stable hydrogen-isotope values of ground water (δ 2 H) and dissolved hydrogen concentrations (H (2(aq) ) were quantified in a petroleum-hydrocarbon contaminated aquifer to determine whether the production/consumption of H 2 by subsurface microorganisms affects ground water &delta 2 H values. The range of &delta 2 H observed in monitoring wells sampled (-27.8 &permil; c to -15.5 &permil; c ) was best explained, however, by seasonal differences in recharge temperature as indicated using ground water δ 18 O values, rather than isotopic exchange reactions involving the microbial cycling of H 2 during anaerobic petroleum-hydrocarbon biodegradation. The absence of a measurable hydrogen-isotope exchange between microbially cycled H 2 and ground water reflects the fact that the amount of H 2 available from the anaerobic decomposition of petroleum hydrocarbons is small relative to the amount of hydrogen present in water, even though milligram per liter concentrations of readily biodegradable contaminants are present at the study site. Additionally, isotopic fractionation calculations indicate that in order for H 2 cycling processes to affect δ 2 H values of ground water, relatively high concentrations of H 2 (>0.080 M) would have to be maintained, considerably higher than the 0.2 to 26 nM present at this site and characteristic of anaerobic conditions in general. These observations suggest that the conventional approach of using δ 2 H and δ 18 O values to determine recharge history is appropriate even for those ground water systems characterized by anaerobic conditions and extensive microbial H 2 cycling.

Ground Water↗

USGS2001

Potential contribution of microbial degradation to natural attenuation of MTBE in surface water systems

The potential contribution of in situ biodegradation as a mechanism for natural attenuation of MTBE in surface water was studied. Surface water sediments from streams and lakes at 11 sites throughout the US. Microbial degradation of [U-14C] MTBE was observed in surface-water-sediment microcosms under anaerobic conditions, but the efficiency and products of anaerobic MTBE biodegradation were strongly dependent on the predominant terminal electron accepting conditions. In the presence of substantial methanogenic activity, MTBE biodegradation was nominal and involved reduction of MTBE to t-butanol (TBA). Under more oxidizing conditions, minimal accumulation of 14C-TBA and significant mineralization of [U-14C] MTBE to 14CO2 were observed. Microorganisms inhabiting the bed sediments of streams and lakes could degrade MTBE effectively under a range of anaerobic terminal electron accepting conditions. Thus, anaerobic bed sediment microbial processes also might contribute to natural attenuation of MTBE in surface water systems throughout the US. This is an abstract of a paper presented at the 222nd ACS National Meting (Chicago, IL 8/26-30/2001).

ACS Division of Environmental Chemistry, Preprints↗

USGS1993

Geobacter metallireducens gen. nov. sp. nov., a microorganism capable of coupling the complete oxidation of organic compounds to the reduction of iron and other metals

The gram-negative metal-reducing microorganism, previously known as strain GS-15, was further characterized. This strict anaerobe oxidizes several short-chain fatty acids, alcohols, and monoaromatic compounds with Fe(III) as the sole electron acceptor. Furthermore, acetate is also oxidized with the reduction of Mn (IV), U (VI), and nitrate. In whole cell suspensions, the c -type cytochrome(s) of this organism was oxidized by physiological electron acceptors and also by gold, silver, mercury, and chromate. Menaquinone was recovered in concentrations comparable to those previously found in gram-negative sulfate reducers. Profiles of the phospholipid ester-linked fatty acids indicated that both the anaerobic desaturase and the branched pathways for fatty acid biosynthesis were operative. The organism contained three lipopolysaccharide hydroxy fatty acids which have not been previously reported in microorganisms, but have been observed in anaerobic freshwater sediments. The 16S rRNA sequence indicated that this organism belongs in the delta proteobacteria. Its closest known relative is Desulfuromonas acetoxidans . The name Geobacter metallireducens is proposed.

Archives of Microbiology↗

USGS2000

Microbial degradation of chloroethenes in groundwater systems

The chloroethenes, tetrachloroethene (PCE) and trichloroethene (TCE) are among the most common contaminants detected in groundwater systems. As recently as 1980, the consensus was that chloroethene compounds were not significantly biodegradable in groundwater. Consequently, efforts to remediate chloroethene-contaminated groundwater were limited to largely unsuccessful pump-and-treat attempts. Subsequent investigation revealed that under reducing conditions, aquifer microorganisms can reductively dechlorinate PCE and TCE to the less chlorinated daughter products dichloroethene (DCE) and vinyl chloride (VC). Although recent laboratory studies conducted with halorespiring microorganisms suggest that complete reduction to ethene is possible, in the majority of groundwater systems reductive dechlorination apparently stops at DCE or VC. However, recent investigations conducted with aquifer and stream-bed sediments have demonstrated that microbial oxidation of these reduced daughter products can be significant under anaerobic redox conditions. The combination of reductive dechlorination of PCE and TCE under anaerobic conditions followed by anaerobic microbial oxidation of DCE and VC provides a possible microbial pathway for complete degradation of chloroethene contaminants in groundwater systems.

Hydrogeology Journal↗