Mark A. Widdowson

Effect of biostimulants on 2,4,6-trinitrotoluene (TNT) degradation and bacterial community composition in contaminated aquifer sediment enrichments

Abstract2,4,6-Trinitrotoluene (TNT) is a toxic and persistent explosive compound occurring as a contaminant at numerous sites worldwide. Knowledge of the microbial dynamics driving TNT biodegradation is limited, particularly in native aquifer sediments where it poses a threat to water resources. The purpose of this study was to quantify the effect of organic amendments on anaerobic TNT biodegradation rate and pathway in an enrichment culture obtained from historically contaminated aquifer sediment and to compare the bacterial community dynamics. TNT readily biodegraded in all microcosms, with the highest biodegradation rate obtained under the lactate amended condition followed by ethanol amended and naturally occurring organic matter (extracted from site sediment) amended conditions. Although a reductive pathway of TNT degradation was observed across all conditions, denaturing gradient gel electrophoresis (DGGE) analysis revealed distinct bacterial community compositions. In all microcosms, Gram-negative γ- or β-Proteobacteria and Gram-positive Negativicutes or Clostridia were observed. A Pseudomonas sp. in particular was observed to be stimulated under all conditions. According to non-metric multidimensional scaling analysis of DGGE profiles, the microcosm communities were most similar to heavily TNT-contaminated field site sediment, relative to moderately and uncontaminated sediments, suggesting that TNT contamination itself is a major driver of microbial community structure. Overall these results provide a new line of evidence of the key bacteria driving TNT degradation in aquifer sediments and their dynamics in response to organic carbon amendment, supporting this approach as a promising technology for stimulating in situ TNT bioremediation in the subsurface.

Modeling Natural Attenuation of Chlorinated Ethenes Under Spatially Varying Redox Conditions

A three-dimensional model for the transport and reductive dechlorination of chlorinated ethenes in ground-water systems with variable redox conditions is demonstrated and applied to a pilot test for accelerated natural attenuation of trichloroethene (TCE). The rate and extent of biotransformation of TCE and chlorinated progeny is controlled by the dominant terminal electron accepting process (TEAP) that is simulated over space and time. The solute transport code, Sequential Electron Acceptor Model, 3D-transport, (SEAM3D) which simulates aerobic and sequential anaerobic biodegradation of organic carbon, is modified to implement the equations. Results of a generic model for TCE transport in ground-water systems with different redox conditions demonstrate that the degree of chlorinated ethene attenuation is influenced by background concentrations of aqueous- and solid-phase electron acceptors, but that model results are sensitive to other input parameters (inhibition coefficients, maximum rate of reductive dechlorination, biomass concentrations, and ground-water velocity). Simulation results of enhanced in situ bioremediation using dissolved organic carbon as a reducing agent show that spatial and temporal changes in the dominant TEAP and the subsequent rate of reductive dechlorination are adequately represented with the model. Initial concentrations of Fe(III) and the dechlorinating microbial population influence the simulated time lag observed during the pilot test.

Controls on mixing‐dependent denitrification in hyporheic zones induced by riverbed dunes: A steady state modeling study

The hyporheic zone is known to attenuate contaminants originating from surface water, yet the ability of the hyporheic zone to attenuate contaminants in upwelling groundwater plumes as they exit to surface water is less understood. We used MODFLOW and SEAM3D to simulate hyporheic flow cells induced by riverbed dunes and upwelling groundwater together with mixing-dependent denitrification of an upwelling nitrate ( NO3−) plume. Our base case modeled labile dissolved organic carbon (DOC) and dissolved oxygen (DO) advecting from surface water, and DO and NO3− advecting from groundwater, typical of certain agricultural areas. We conducted sensitivity analyses that showed mixing-dependent denitrification in the hyporheic zone increased with increasing hydraulic conductivity (K), decreasing lower boundary flux, and increasing DOC in surface water or NO3− in groundwater. Surface water DOC, groundwater NO3−, and K were the most sensitive parameters affecting mixing-dependent denitrification. Nonmixing-dependent denitrification also occurred when there was surface water NO3−, and its magnitude was often greater than mixing-dependent denitrification. Nevertheless, mixing-dependent reactions provide functions that nonmixing-dependent reactions cannot, with potential for hyporheic zones to attenuate upwelling NO3− plumes, depending on geomorphic, hydraulic, and biogeochemical conditions. Stream and river restoration efforts may be able to increase mixing-dependent reactions by promoting natural processes that promote bedform creation and augment labile carbon sources.

Biodegradation of a PAH Mixture by Native Subsurface Microbiota

Laboratory microcosm studies were conducted to estimate biodegradation rates for a mixture of five polycyclic aromatic hydrocarbon compounds (PAHs). Static microcosms were assembled using soil samples from two locations collected at a No. 2 fuel oil-contaminated site in the Atlantic Coastal Plain of Virginia. In microcosms from one location, five PAHs (acenaphthene, fluorene, phenanthrene, pyrene, and benzo(b)fluoranthene) biodegraded at net first-order rates of 1.08, 1.45, 1.13, 1.11, and 1.12 yr−1, respectively. No observable lag period was noted and degradation in live microcosms ceased with the depletion of oxygen and sulfate after 125 days. In microcosms from a second location, net first-order biodegradation rates after an approximately 2-month lag period were 2.41, 3.28, and 2.98 yr−1 for fluorene, phenanthrene, and pyrene, respectively. Acenaphthene and benzo(b)fluoranthene mass loss rates in the live microcosms were not statistically different from mass loss rates in control microcosms. Stoichiometric mass balance calculations indicate that the dominant PAH mass loss mechanism was aerobic biodegradation, while abiotic losses (attributed to micropore diffusion and oxidative coupling) ranged from 15 to 33% and biotic losses from sulfate-reduction accounted for 7 to 10% of PAH mass loss. Stoichiometric equations that include biomass yield are presented for PAH oxidation under aerobic and sulfate-reducing conditions.

Simplified methods for monitoring petroleum‐contaminated ground water and soil vapor

Portable meters and simplified gas Chromatographic (GC) techniques were investigated for monitoring volatile hydrocarbon (HC), CO2, and O2, concentrations in groundwater, exhaust gases, and soil vapor during in situ remediation using soil vapor extraction (SVE) and air sparging (AS). Results of groundwater samples analyzed in‐house using a headspace technique compared well to split samples analyzed by a certified analytical laboratory (r2 = 0.94). SVE exhaust gas HC and CO2 concentrations measured using a GT201 portable HC/O2 meter and a RA‐411A meter (GasTech), respectively, were highly correlated with in‐house laboratory GC analyses (r2 = 0.91). O2 concentrations fell in a small range and meter analyses were not well correlated with laboratory analyses. Results of soil gas monitoring were not as well correlated as those for exhaust gases for HC, CO2, or O2, perhaps due to environmental conditions such as changes in relative humidity or the wider range of soil gas values. Overall, the meters were good in...

In-situ determination of field-scale NAPL mass transfer coefficients: Performance, simulation and analysis

Better estimates of non-aqueous phase liquid (NAPL) mass, its persistence into the future, and the potential impact of source reduction are critical needs for determining the optimal path to clean up sites impacted by NAPLs. One impediment to constraining time estimates of source depletion is the uncertainty in the rate of mass transfer between NAPLs and groundwater. In this study, an innovative field test is demonstrated for the purpose of quantifying field-scale NAPL mass transfer coefficients (kl(N)) within a source zone of a fuel-contaminated site. Initial evaluation of the test concept using a numerical model revealed that the aqueous phase concentration response to the injection of clean groundwater within a source zone was a function of NAPL mass transfer. Under rate limited conditions, NAPL dissolution together with the injection flow rate and the radial distance to monitoring points directly controlled time of travel. Concentration responses observed in the field test were consistent with the hypothetical model results allowing field-scale NAPL mass transfer coefficients to be quantified. Site models for groundwater flow and solute transport were systematically calibrated and utilized for data analysis. Results show kl(N) for benzene varied from 0.022 to 0.60d(-1). Variability in results was attributed to a highly heterogeneous horizon consisting of layered media of varying physical properties.

Multicomponent NAPL Source Dissolution: Evaluation of Mass-Transfer Coefficients

Mass transfer rate coefficients were quantified by employing an inverse modeling technique to high-resolution aqueous phase concentration data observed following an experimental release of a multicomponent nonaqueous phase liquid (NAPL) at a field site. A solute transport model (SEAM3D) was employed to simulate advective-dispersive transport over time coupled to NAPL dissolution. Model calibration was demonstrated by accurately reproducing the observed breakthrough times and peak concentrations at multiple observation points, observed mass discharge at pumping wells, and the reported mass depletions for three soluble NAPL constituents. Vertically variable NAPL mass transfer coefficients were derived for each constituent using an optimized numerical solute transport model, ranging from 0.082 to 2.0 day(-1) across all constituents. Constituent-specific coefficients showed a positive correlation with liquid-phase diffusion coefficients. Application of a time-varying mass transfer coefficient as NAPL mass depleted showed limited sensitivity during which over 80% of the most soluble NAPL constituent dissolved from the source. Long-term simulation results, calibrated to the experimental data and rendered in terms of mass discharge versus source mass depletion, exhibited multistage behavior.

Stimulating In Situ Hydrogenotrophic Denitrification with Membrane-Delivered Hydrogen under Passive and Pumped Groundwater Conditions

A technology was developed to stimulate autotrophic biological denitrification by supplying hydrogen ( H2 ) to groundwater via gas-permeable membranes. The purpose of this project was to investigate this technology at field scale, determining whether it could be successfully scaled up from the laboratory. The field site was located in Becker, Minnesota and contained high levels of NO 3− ( 22.8±2.0 mg/L-N ) and dissolved oxygen (DO) ( 7±1 mg/L ) . Membranes installed in groundwater wells were successful in delivering H2 to the groundwater over the two-year operating period. Hydrogen stimulated microbial reduction of DO and NO 3− , degrading up to 6 mg/L DO and converting up to 10.0 mg/L NO 3− -N to NO 2− -N when operated passively. When recirculation pumps were installed performance in the field did not improve significantly because of mixing with more oxygenated water. However, complementary modeling studies showed that complete DO reduction and denitrification to N2 was possible but the zone of influence...

Sequential Electron Acceptor Model for Evaluation of in Situ Bioremediation of Petroleum Hydrocarbon Contaminants in Groundwatera

Mathematical development and model application is provided for a multiple substrate, sequential electron acceptor model, accounting for hydrodynamic transport, adsorption, and sequential oxygen/iron(III)-based biodegradation. Equations for iron(III)-based biodegradation of petroleum hydrocarbons are developed based on oxygen-inhibited Monod kinetics. The iron(III)-based biodegradation expressions were combined with earlier work by Widdowson and Aelion, to develop the two-dimensional, multiple substrate, oxygen/iron(III) sequential electron acceptor biodegradation model presented here. In addition to mathematical model development, simulations demonstrating the advantages of sequential electron acceptor and multiple substrate biodegradation models are provided. These simulations show that commonly-used single electron acceptor models may underpredict natural, in situ biodegradation potential at sites where indigenous microorganisms are capable of using multiple electron acceptors. Additional simulations show that, for contaminant plumes composed of constituents which biodegrade at different rates and under varying electron acceptor conditions, a multiple substrate model may allow more accurate prediction of both individual contaminant concentrations and the total amount of biodegraded contaminant. Considering that typical contaminant plumes are composed of multiple constituents with varying biodegradation properties and health risks, multiple substrate sequential electron acceptor models have the potential to provide more accurate tracking of individual constituent migration. The model was applied to a leaking UST site in Laurel Bay, South Carolina. Laboratory and monitoring well data presented in Landmeyer et al. have established that the petroleum hydrocarbon contaminants are present in the groundwater and are undergoing sequential oxygen-iron(III)-based biodegradation. Model simulations proved capable of reproducing the trends observed at the Laurel Bay site in that BTX contaminants were removed by sequential biodegradation, occurring first aerobically and subsequently anaerobically, and that iron(III)-reducing organisms biodegrade contaminants only in the absence of oxygen. The BTX compounds were individually but simultaneously modeled, allowing explicit modeling of specific contaminant biodegradation properties (e.g., toluene and xylene are assumed to degrade sequentially and benzene is assumed to degrade aerobically only). Although simulations presented here can reproduce trends observed at the Laurel Bay site, inclusion of additional electron acceptors and additional model calibration to data from this and other sites is necessary to improve and verify the models capability to predict the efficacy of intrinsic biodegradation of petroleum hydrocarbon contaminants in groundwater.

Kinetic and Pathway Modeling of Reductive 2,4,6-Trinitrotoluene Biodegradation with Different Electron Donors

AbstractA comprehensive model was applied to simulate a laboratory microcosm study of biodegradation rates and the branched production and loss of daughter products. The aim of the investigation was to evaluate the effect of electron donors (lactate, ethanol, and natural organic matter) on 2,4,6-trinitrotoluene (TNT) biodegradation rate and pathway in historically contaminated sediments undergoing biostimulation. Simulation results show overall TNT degradation rates for lactate-amended microcosms were greater than ethanol-amended microcosms by a factor of 1.7 and 3.0 times compared with natural organic matter amended microcosms. Differences in observed biomass concentrations (lactate>ethanol>unamended) were thought to be a contributing factor. TNT degradation pathway modeling included determination of branching coefficients representing whether the first nitro group reduction occurred in the ortho or para position. Branching coefficients were greater for the initial reduction of para (17–27% initial TNT c...