On the reliable estimation of sequential Monod kinetic parameters.

While dozens of studies have attempted to estimate the Monod kinetic parameters of microbial reductive dechlorination, published values in the literature vary by 2-6 orders of magnitude. This lack of consensus can be attributed in part to limitations of both experimental design and parameter estimation techniques. To address these issues, Hamiltonian Monte Carlo was used to produce more than one million sets of realistic simulated microcosm data under a variety of experimental conditions. These data were then employed in model fitting experiments using a number of parameter estimation algorithms for determining Monod kinetic parameters. Analysis of data from conventional triplicate microcosms yielded parameter estimates characterized by high collinearity, resulting in poor estimation accuracy and precision. Additionally, confidence intervals computed by commonly used classical regression analysis techniques contained true parameter values much less frequently than their nominal confidence levels. Use of an alternative experimental design, requiring the same number of analyses as conventional experiments but comprised of microcosms with varying initial chlorinated ethene concentrations, is shown to result in order-of-magnitude decreases in parameter uncertainty. A Metropolis algorithm which can be run on a typical personal computer is demonstrated to return more reliable parameter interval estimates.

Quantifying Impacts of Microcosm Mass Loss on Kinetic Constant Estimation.

Microcosm experiments to assess microbial reductive dechlorination of chlorinated aliphatic hydrocarbons typically experience 5-50% mass loss due to frequent sampling events and diffusion through septa. A literature review, however, reveals that models fit to such experiments for kinetic constant estimation have generally failed to account for experimental mass loss. To investigate possible resultant bias in best-fit parameters, a series of numerical experiments was conducted in which Monod kinetic models with and without mass loss were fit to more than 1300 synthetic data sets, generated using published microcosm data. Models that failed to account for mass loss resulted in significant fitted parameter bias. Bias ranged from 5 to 45% of the parameter magnitude for Monte Carlo simulations with low (approximately 10%) mass loss to 20-120% for simulations with high (approximately 40%) mass loss. In addition, for high mass loss simulations, best-fit values consistently fell along the bounds of the optimization range. These results suggest that failure to properly account for mass loss in microcosms may lead to inaccurate estimation of kinetic constants and may explain some of the literature-reported variability in these parameters. A model is presented that provides a method for including sampling and diffusional mass losses to improve kinetic constant estimation accuracy.

Exploration of processes governing microbial reductive dechlorination in a heterogeneous aquifer flow cell.

Although microbial reductive dechlorination (MRD) has proven to be an effective approach for in situ treatment of chlorinated ethenes, field implementation of this technology is complicated by many factors, including subsurface heterogeneity, electron donor availability, and distribution of microbial populations. This work presents a coupled experimental and mathematical modeling study designed to explore the influence of heterogeneity on MRD and to assess the suitability of microcosm-derived rate parameters for modeling complex heterogeneous systems. A Monod-based model is applied to simulate a bioremediation experiment conducted in a laboratory-scale aquifer cell packed with aquifer material from the Commerce Street Superfund site in Williston, VT. Results reveal that (uncalibrated) model application of microcosm-derived dechlorination and microbial growth rates for transformation of trichloroethene (TCE), cis-1,2-dichloroethene (cis-DCE), and vinyl chloride (VC) reproduced observed aquifer cell concentration levels and trends. Mean relative errors between predicted and measured effluent concentrations were quantified as 6.7%, 27.0%, 41.5%, 32.0% and 21.6% over time for TCE, cis-DCE, VC, ethene and total volatile fatty acids (fermentable electron donor substrate and carbon source), respectively. The time-averaged extent of MRD (i.e., ethene formation) was well-predicted (4% underprediction), with modeled MRD exhibiting increased deviation from measured values under electron donor limiting conditions (maximum discrepancy of 14%). In contrast, simulations employing a homogeneous (uniform flow) domain resulted in underprediction of MRD extent by an average of 13%, with a maximum discrepancy of 45%. Model sensitivity analysis suggested that trace amounts of natural dissolved organic carbon served as an important fermentable substrate, providing up to 69% of the reducing equivalents consumed for MRD under donor-limiting conditions. Aquifer cell port concentration data and model simulations revealed that ethene formation varied spatially within the domain and was associated with regions of longer residence times. These results demonstrate the strong influence of subsurface heterogeneity on the accuracy of MRD predictions, and highlight the importance of subsurface characterization and the incorporation of flow field uncertainty in model applications for successful design and assessment of in situ bioremediation.

Development and application of a screening model for evaluating bioenhanced dissolution in DNAPL source zones.

In-situ bioremediation, a widely applied treatment technology for source zones contaminated with dense non-aqueous phase liquids (DNAPLs), has proven economical and reasonably efficient for long-term management of contaminated sites. Successful application of this remedial technology, however, requires an understanding of the complex interaction of transport, mass transfer, and biotransformation processes. The bioenhancement factor, which represents the ratio of DNAPL mass transfer under microbially active conditions to that which would occur under abiotic conditions, is commonly used to quantify the effectiveness of a particular bioremediation remedy. To date, little research has been directed towards the development and validation of methods to predict bioenhancement factors under conditions representative of real sites. This work extends an existing, first-order, bioenhancement factor expression to systems with zero-order and Monod kinetics, representative of many source-zone scenarios. The utility of this model for predicting the bioenhancement factor for previously published laboratory and field experiments is evaluated. This evaluation demonstrates the applicability of these simple bioenhancement factors for preliminary experimental design and analysis, and for assessment of dissolution enhancement in ganglia-contaminated source zones. For ease of application, a set of nomographs is presented that graphically depicts the dependence of bioenhancement factor on physicochemical properties. Application of these nomographs is illustrated using data from a well-documented field site. Results suggest that this approach can successfully capture field-scale, as well as column-scale, behavior. Sensitivity analyses reveal that bioenhanced dissolution will critically depend on in-situ biomass concentrations.

Effect of NAPL Source Morphology on Mass Transfer in the Vadose Zone.

The generation of vapor-phase contaminant plumes within the vadose zone is of interest for contaminated site management. Therefore, it is important to understand vapor sources such as non-aqueous-phase liquids (NAPLs) and processes that govern their volatilization. The distribution of NAPL, gas, and water phases within a source zone is expected to influence the rate of volatilization. However, the effect of this distribution morphology on volatilization has not been thoroughly quantified. Because field quantification of NAPL volatilization is often infeasible, a controlled laboratory experiment was conducted in a two-dimensional tank (28 cm × 15.5 cm × 2.5 cm) with water-wet sandy media and an emplaced trichloroethylene (TCE) source. The source was emplaced in two configurations to represent morphologies encountered in field settings: (1) NAPL pools directly exposed to the air phase and (2) NAPLs trapped in water-saturated zones that were occluded from the air phase. Airflow was passed through the tank and effluent concentrations of TCE were quantified. Models were used to analyze results, which indicated that mass transfer from directly exposed NAPL was fast and controlled by advective-dispersive-diffusive transport in the gas phase. However, sources occluded by pore water showed strong rate limitations and slower effective mass transfer. This difference is explained by diffusional resistance within the aqueous phase. Results demonstrate that vapor generation rates from a NAPL source will be influenced by the soil water content distribution within the source. The implications of the NAPL morphology on volatilization in the context of a dynamic water table or climate are discussed.

Microbially enhanced dissolution and reductive dechlorination of PCE by a mixed culture: model validation and sensitivity analysis.

Reductive dechlorination catalyzed by organohalide-respiring bacteria is often considered for remediation of non-aqueous phase liquid (NAPL) source zones due to cost savings, ease of implementation, regulatory acceptance, and sustainability. Despite knowledge of the key dechlorinators, an understanding of the processes and factors that control NAPL dissolution rates and detoxification (i.e., ethene formation) is lacking. A recent column study demonstrated a 5-fold cumulative enhancement in tetrachloroethene (PCE) dissolution and ethene formation (Amos et al., 2009). Spatial and temporal monitoring of key geochemical and microbial (i.e., Geobacter lovleyi and Dehalococcoides mccartyi strains) parameters in the column generated a data set used herein as the basis for refinement and testing of a multiphase, compositional transport model. The refined model is capable of simulating the reactive transport of multiple chemical constituents produced and consumed by organohalide-respiring bacteria and accounts for substrate limitations and competitive inhibition. Parameter estimation techniques were used to optimize the values of sensitive microbial kinetic parameters, including maximum utilization rates, biomass yield coefficients, and endogenous decay rates. Comparison and calibration of model simulations with the experimental data demonstrate that the model is able to accurately reproduce measured effluent concentrations, while delineating trends in dechlorinator growth and reductive dechlorination kinetics along the column. Sensitivity analyses performed on the optimized model parameters indicate that the rates of PCE and cis-1,2-dichloroethene (cis-DCE) transformation and Dehalococcoides growth govern bioenhanced dissolution, as long as electron donor (i.e., hydrogen flux) is not limiting. Dissolution enhancements were shown to be independent of cis-DCE accumulation; however, accumulation of cis-DCE, as well as column length and flow rate (i.e., column residence time), strongly influenced the extent of reductive dechlorination. When cis-DCE inhibition was neglected, the model over-predicted ethene production ten-fold, while reductions in residence time (i.e., a two-fold decrease in column length or two-fold increase in flow rate) resulted in a more than 70% decline in ethene production. These results suggest that spatial and temporal variations in microbial community composition and activity must be understood to model, predict, and manage bioenhanced NAPL dissolution.

Degradation product partitioning in source zones containing chlorinated ethene dense non-aqueous-phase liquid.

Abiotic and biotic reductive dechlorination with chlorinated ethene dense non-aqueous-phase liquid (DNAPL) source zones can lead to significant fluxes of complete and incomplete transformation products. Accurate assessment of in situ rates of transformation and the potential for product sequestration requires knowledge of the distribution of these products among the solid, aqueous, and organic liquid phases present within the source zone. Here we consider the fluid-fluid partitioning of two of the most common incomplete transformation products, cis-1,2-dichloroethene (cis-DCE) and vinyl chloride (VC). The distributions of cis-DCE and VC between the aqueous phase and tetrachloroethene (PCE) and trichloroethene (TCE) DNAPLs, respectively, were quantified at 22 °C for the environmentally relevant, dilute range. The results suggest that partition coefficients (concentration basis) for VC and cis-DCE are 70 ± 1 L(aq)/L(TCE DNAPL) and 105 ± 1 L(aq)/L(PCE DNAPL,) respectively. VC partitioning data (in the dilute region) were reasonably approximated using the Raoult's law analogy for liquid-liquid equilibrium. In contrast, data for the partitioning of cis-DCE were well described only when well-parametrized models for the excess Gibbs free energy were employed. In addition, available vapor-liquid and liquid-liquid data were employed with our measurements to assess the temperature dependence of the cis-DCE and VC partition coefficients. Overall, the results suggest that there is a strong thermodynamic driving force for the reversible sequestration of cis-DC and VC within DNAPL source zones. Implications of this partitioning include retardation during transport and underestimation of the transformation rates observed through analysis of aqueous-phase samples.

Predicting DNAPL mass discharge from pool-dominated source zones.

Models that link simplified descriptions of dense non-aqueous phase liquid (DNAPL) source zone architecture with predictions of mass flux can be effective screening tools for evaluation of source zone management strategies. Recent efforts have focused on the development and implementation of upscaled models to approximate the relationship between mass removal and flux-averaged, down-gradient contaminant concentration (or mass flux) reduction. The efficacy of these methods has been demonstrated for ganglia-dominated source zones. This work extends these methods to source zones dominated by high-saturation DNAPL pools. An existing upscaled mass transfer model was modified to reproduce dissolution behavior in pool-dominated scenarios by employing a two-domain (ganglia and pools) representation of the source zone. The two-domain upscaled model is parameterized using the initial fraction of the source zone that exists as pool regions, the initial fraction of contaminant eluting from these pool regions, and the flux-averaged down-gradient contaminant concentration. Comparisons of model predictions with a series of three-dimensional source zone numerical simulations and data from two-dimensional aquifer cell experiments demonstrate the ability of the model to predict DNAPL dissolution from ganglia- and pool-dominated source zones for all levels of mass recovery.

An assembly model for simulation of large-scale ground water flow and transport.

When managing large-scale ground water contamination problems, it is often necessary to model flow and transport using finely discretized domains--for instance (1) to simulate flow and transport near a contamination source area or in the area where a remediation technology is being implemented; (2) to account for small-scale heterogeneities; (3) to represent ground water-surface water interactions; or (4) some combination of these scenarios. A model with a large domain and fine-grid resolution will need extensive computing resources. In this work, a domain decomposition-based assembly model implemented in a parallel computing environment is developed, which will allow efficient simulation of large-scale ground water flow and transport problems using domain-wide grid refinement. The method employs common ground water flow (MODFLOW) and transport (RT3D) simulators, enabling the solution of almost all commonly encountered ground water flow and transport problems. The basic approach partitions a large model domain into any number of subdomains. Parallel processors are used to solve the model equations within each subdomain. Schwarz iteration is applied to match the flow solution at the subdomain boundaries. For the transport model, an extended numerical array is implemented to permit the exchange of dispersive and advective flux information across subdomain boundaries. The model is verified using a conventional single-domain model. Model simulations demonstrate that the proposed model operated in a parallel computing environment can result in considerable savings in computer run times (between 50% and 80%) compared with conventional modeling approaches and may be used to simulate grid discretizations that were formerly intractable.

Experimental evaluation and mathematical modeling of microbially enhanced tetrachloroethene (PCE) dissolution.

Experiments to assess metabolic reductive dechlorination (chlororespiration) at high concentration levels consistent with the presence of free-phase tetrachloroethene (PCE) were performed using three PCE-to-cis-1,2-dichloroethene (cis-DCE) dechlorinating pure cultures (Sulfurospirillum multivorans, Desulfuromonas michiganensis strain BB1, and Geobacter lovleyi strain SZ) and Desulfitobacterium sp. strain Viet1, a PCE-to-trichloroethene (TCE) dechlorinating isolate. Despite recent evidence suggesting bacterial PCE-to-cis-DCE dechlorination occurs at or near PCE saturation (0.9-1.2 mM), all cultures tested ceased dechlorinating at approximately 0.54 mM PCE. In the presence of PCE dense nonaqueous phase liquid (DNAPL), strains BB1 and SZ initially dechlorinated, but TCE and cis-DCE production ceased when aqueous PCE concentrations reached inhibitory levels. For S. multivorans, dechlorination proceeded at a rate sufficient to maintain PCE concentrations below inhibitory levels, resulting in continuous cis-DCE production and complete dissolution of the PCE DNAPL. A novel mathematical model, which accounts for loss of dechlorinating activity at inhibitory PCE concentrations, was developed to simultaneously describe PCE-DNAPL dissolution and reductive dechlorination kinetics. The model predicted that conditions corresponding to a bioavailability number (Bn) less than 1.25 x 10(-2) will lead to dissolution enhancement with the tested cultures, while conditions corresponding to a Bn greater than this threshold value can result in accumulation of PCE to inhibitory dissolved-phase levels, limiting PCE transformation and dissolution enhancement. These results suggest that microorganisms incapable of dechlorinating at high PCE concentrations can enhance the dissolution and transformation of PCE from free-phase DNAPL.

Coupling aggressive mass removal with microbial reductive dechlorination for remediation of DNAPL source zones: a review and assessment.

The infiltration of dense non-aqueous-phase liquids (DNAPLs) into the saturated subsurface typically produces a highly contaminated zone that serves as a long-term source of dissolved-phase groundwater contamination. Applications of aggressive physical-chemical technologies to such source zones may remove > 90% of the contaminant mass under favorable conditions. The remaining contaminant mass, however, can create a rebounding of aqueous-phase concentrations within the treated zone. Stimulation of microbial reductive dechlorination within the source zone after aggressive mass removal has recently been proposed as a promising staged-treatment remediation technology for transforming the remaining contaminant mass. This article reviews available laboratory and field evidence that supports the development of a treatment strategy that combines aggressive source-zone removal technologies with subsequent promotion of sustained microbial reductive dechlorination. Physical-chemical source-zone treatment technologies compatible with posttreatment stimulation of microbial activity are identified, and studies examining the requirements and controls (i.e., limits) of reductive dechlorination of chlorinated ethenes are investigated. Illustrative calculations are presented to explore the potential effects of source-zone management alternatives. Results suggest that, for the favorable conditions assumed in these calculations (i.e., statistical homogeneity of aquifer properties, known source-zone DNAPL distribution, and successful bioenhancement in the source zone), source longevity may be reduced by as much as an order of magnitude when physical-chemical source-zone treatment is coupled with reductive dechlorination.

Effects of carbonate species on the kinetics of dechlorination of 1,1,1-trichloroethane by zero-valent iron.

The effect of precipitates on the reactivity of iron metal (Fe0) with 1,1,1-trichloroethane (TCA) was studied in batch systems designed to model groundwaters that contain dissolved carbonate species (i.e., C(IV)). At representative concentrations for high-C(IV) groundwaters (approximately 10(-2) M), the pH in batch reactors containing Fe0 was effectively buffered until most of the aqueous C(IV) precipitated. The precipitate was mainly FeCO3 (siderite) but may also have included some carbonate green rust. Exposure of the Fe0 to dissolved C(IV) accelerated reduction of TCA, and the products formed under these conditions consisted mainly of ethane and ethene, with minor amounts of several butenes. The kinetics of TCA reduction were first-order when C(IV)-enhanced corrosion predominated but showed mixed-order kinetics (zero- and first-order) in experiments performed with passivated Fe0 (i.e., before the onset of pitting corrosion and after repassivation by precipitation of FeCO3). All these data were described by fitting a Michaelis-Menten-type kinetic model and approximating the first-order rate constant as the ratio of the maximum reaction rate (Vm) and the concentration of TCA at half of the maximum rate (K(1/2)). The decrease in Vm/K(1/2) with increasing C(IV) exposure time was fit to a heuristic model assuming proportionality between changes in TCA reduction rate and changes in surface coverage with FeCO3.