Reductive dechlorination of the vinyl chloride surrogate chlorofluoroethene in TCE-contaminated groundwater

At many trichloroethene (TCE)-contaminated field sites, microbial transformation of TCE results in the accumulation of vinyl chloride (VC), a known carcinogen and neurotoxin. Quantitative tools are needed to determine the in situ rates of VC transformation to ethene in contaminated groundwater. For this study, E-/Z-chlorofluoroethene (E-/Z-CFE) was evaluated as a surrogate for VC in laboratory microcosm and field push-pull tests. Single-well push-pull tests were conducted at a TCE-contaminated field site by injecting E-/Z-CFE and monitoring for the formation of fluoroethene (FE) over a period of up to 80 days. The rates for VC transformation to ethene and E-CFE transformation to FE were within a factor of 2.7 for laboratory microcosm systems and all preferentiallytransformed E-CFE over Z-CFE. In the field, the in situ rates of FE production from injected E-CFE ranged from 0.0018 to 1.15 microM/day, while the in situ rates of E-CFE disappearance ranged from 0.17 to 0.99 microM/day. No significant Z-CFE transformation was observed in field tests, which indicated preferential utilization of E-CFE over Z-CFE under in situ field conditions. The results of this study indicate E-CFE as a potential surrogate for estimating the in situ rates of VC transformation.     

"Forced mass balance" technique for estimating in situ transformation rates of sorbing solutes in groundwater

A method for estimating in situ transformation rates of sorbing solutes in groundwater is presented. The method utilizes a novel data processing technique called "forced mass balance" (FMB) to remove the effects of transport processes from reactant and product concentrations measured during single-well, "push-pull" tests. The effectiveness of the FMB technique was evaluated by quantifying errors in derived rates obtained by applying FMB to simulated push-pull test data generated by a numerical model. Results from simulated tests indicated that errors in derived rates increase as the test duration, groundwater velocity, and ratio of reactant to product retardation factors increase. In addition, errors in derived rates increase as the reaction rate constant and aquifer dispersivity decrease. As a demonstration, the FMB technique was used to derive an in situ reductive dechlorination rate for trichlorofluoroethene (TCFE) using data from a field push-pull test. Error analyses indicated that the in situ TCFE transformation rate was underestimated by a factor of 1.1-2. Thus, the FMB technique makes it possible to estimate in situ transformation rates of sorbing solutes and when FMB is coupled with computer modeling to estimate errors in derived in situ rates.     

Anaerobic reductive dechlorination of 1-chloro-1-fluoroethene to track the transformation of vinyl chloride

1-Chloro-1-fluoroethene (1,1-CFE) was studied as a reactive tracer to quantify the anaerobic transformation of vinyl chloride (VC). Batch kinetic studies of 1,1-CFE and VC transformation were performed with an enrichment culture obtained from the Evanite site in Corvallis, OR. The culture is capable of completely transforming trichloroethene (TCE) through cis-dichloroethene (c-DCE) and VC to ethene. The culture also transforms fluorinated analogues, such as trichlorofluoroethene (TCFE), to fluoroethene (FE) as a final product. The transformation sequence of the fluorinated analogue was correlated with that achieved for the chlorinated ethene with the same degree of chloride substitution. For example, the production of 1,1-CFE, the major CFE isomer formed from TCFE transformation, was correlated with the production of VC from TCE transformation. Since the 1,1-CFE and its product, FE, have a distinct analytical signature, 1,1-CFE may be used as a reactive in situ tracer to evaluate the VC transformation potential. The half-saturated constants (K(S)) of VC and 1,1-CFE were 63 and 87 microM, respectively, while similar maximum utilization rates (kmaxX) of 334 and 350 microM/d were achieved. Acetylene inhibited both VC and 1,1-CFE transformation. A competitive inhibition model with the independently measured K(S) values used as the inhibition constants predicted rates of transformation of both VC and 1,1-CFE when both compounds were present. 1,1-CFE transformation was also tested with three different cultures. With all the cultures, 1,1-CFE transformation was associated with VC transformation to ethene, and the rates of transformation were comparable. The results demonstrated that 1,1-CFE was a good reactive surrogate for evaluating the rates of VC transformation.     

Isotopic and geochemical assessment of in situ biodegradation of chlorinated hydrocarbons

Currently there is no in situ method to detect and quantify complete mineralization of chlorinated hydrocarbons (CHCs) to CO2. Combined isotopic measurements in conjunction with traditional chemical techniques were used to assess in situ biodegradation of trichloroethylene (TCE) and carbon tetrachloride (CT). Vadose zone CHC, ethene, ethane, methane, O2, and CO2 concentrations were analyzed using gas chromatography over 114 days at the Savannah River Site. delta13C of CHC and delta13C and 14C of vadose zone CO2, sediment organic matter, and groundwater dissolved inorganic carbon (DIC)were measured. Intermediate metabolites of TCE and CT accounted for < or = 10% of total CHCs. Delta13C of cis-1,2-dichloroethylene (DCE) was always heavier than TCE indicating substantial DCE biodegradation. 14C-CO2 values ranged from 84 to 128 percent modern carbon (pMC), suggesting that plant root-respired CO2 was dominant. 14C-CO2 values decreased over time (up to 12 pMC), and contaminated groundwater 14C-DIC (76 pMC) was substantially depleted relative to the control (121 pMC). 14C provided a direct measure of complete CHC mineralization in vadose zone and groundwater in situ and may improve remediation strategies.     

In situ stimulation of groundwater denitrification with formate to remediate nitrate contamination

In situ stimulation of denitrification has been proposed as a mechanism to remediate groundwater nitrate contamination. In this study, sodium formate was added to a sand and gravel aquifer on Cape Cod, MA, to test whether formate could serve as a potential electron donor for subsurface denitrification. During 16- and 10-day trials, groundwater from an anoxic nitrate-containing zone (0.5-1.5 mM) was continuously withdrawn, amended with formate and bromide, and pumped back into the aquifer. Concentrations of groundwater constituents were monitored in multilevel samplers after up to 15 m of transport by natural gradient flow. Nitrate and formate concentrations were decreased 80-100% and 60-70%, respectively, with time and subsequent travel distance, while nitrite concentrations inversely increased. The field experiment breakthrough curves were simulated with a two-dimensional site-specific model that included transport, denitrification, and microbial growth. Initial values for model parameters were obtained from laboratory incubations with aquifer core material and then refined to fit field breakthrough curves. The model and the lab results indicated that formate-enhanced nitrite reduction was nearly 4-fold slower than nitrate reduction, but in the lab, nitrite was completely consumed with sufficient exposure time. Results of this study suggest that a long-term injection of formate is necessary to test the remediation potential of this approach for nitrate contamination and that adaptation to nitrite accumulation will be a key determinative factor.     

Field demonstration of DNAPL dehalogenation using emulsified zero-valent iron

This paper describes the results of the first field-scale demonstration conducted to evaluate the performance of nanoscale emulsified zero-valent iron (EZVI) injected into the saturated zone to enhance in situ dehalogenation of dense, nonaqueous phase liquids (DNAPLs) containing trichloroethene (TCE). EZVI is an innovative and emerging remediation technology. EZVI is a surfactant-stabilized, biodegradable emulsion that forms emulsion droplets consisting of an oil-liquid membrane surrounding zero-valent iron (ZVI) particles in water. EZVI was injected over a five day period into eight wells in a demonstration test area within a larger DNAPL source area at NASA's Launch Complex 34 (LC34) using a pressure pulse injection method. Soil and groundwater samples were collected before and after treatment and analyzed for volatile organic compounds (VOCs) to evaluate the changes in VOC mass, concentration and mass flux. Significant reductions in TCE soil concentrations (>80%) were observed at four of the six soil sampling locations within 90 days of EZVI injection. Somewhat lower reductions were observed at the other two soil sampling locations where visual observations suggest that most of the EZVI migrated up above the target treatment depth. Significant reductions in TCE groundwater concentrations (57 to 100%) were observed at all depths targeted with EZVI. Groundwater samples from the treatment area also showed significant increases in the concentrations of cis-1,2-dichloroethene (cDCE), vinyl chloride (VC) and ethene. The decrease in concentrations of TCE in soil and groundwater samples following treatment with EZVI is believed to be due to abiotic degradation associated with the ZVI as well as biodegradation enhanced by the presence of the oil and surfactant in the EZVI emulsion.     

Stable carbon isotope fractionation during enhanced in situ bioremediation of trichloroethene

Time-series stable carbon isotope monitoring of volatile organic compounds (VOCs) atthe Idaho National Engineering and Environmental Laboratory's (INEEL) field site Test Area North (TAN) was conducted during a pilot study to investigate the treatment potential of using lactate to stimulate in situ biologic reductive dechlorination of trichloroethene (TCE). The isotope ratios of TCE and its biodegradation byproducts, cis-dichloroethene (c-DCE), trans-dichloroethene (t-DCE), vinyl chloride (VC), and ethene, in groundwater samples collected during the pilot studywere preconcentrated with a combination of purge-and-trap and cryogenic techniques in order to allow for reproducible isotopic measurements of the low concentrations of these compounds in the samples (down to 0.04 microM, or 5 ppb, of TCE). Compound-specific stable isotope monitoring of chlorinated solvents clearly differentiated between the effects of groundwater transport, dissolution of DNAPL at the source, and enhanced bioremediation. Isotope data from all wells within the zone of lactate influence exhibited large kinetic isotope effects during the reduction of c-DCE to VC and VC to ethene. Despite these large effects, the carbon isotope ratio of ethene in all these wells reached the carbon isotope ratios of the initial dissolved TCE, confirming the complete conversion of dissolved TCEto ethene. Conversely, the carbon isotope ratios of t-DCE were only marginally affected during the study, indicating that minimal biologic degradation of t-DCE was occurring.     

Biotic and abiotic anaerobic transformations of trichloroethene and cis-1,2-dichloroethene in fractured sandstone

A fractured sandstone aquifer at an industrial site in southern California is contaminated with trichloroethene (TCE) and cis-1,2-dichloroethene (cis-DCE) to depths in excess of 244 m. Field monitoring data suggest that TCE is undergoing reduction to cis-DCE and that additional attenuation is occurring. However, vinyl chloride (VC) and ethene have not been detected in significant amounts, so that if transformation is occurring, a process other than reductive dechlorination must be responsible. The objective of this study was to evaluate the occurrence of biotic and abiotic transformation processes at this site for TCE, cis-DCE, and VC. Anaerobic microcosms were constructed with site groundwater and sandstone core samples. 14C-labeled compounds were used to detect transformation products (e.g., CO2 and soluble products) that are not readily identifiable by headspace analysis. The microcosms confirmed the occurrence of biotic reduction of TCE to cis-DCE, driven by electron donor in the groundwater and/or sandstone. VC and ethene were not detected. Following incubation periods up to 22 months, the distribution of 14C indicated statistically significant transformation of [14C]TCE and [14C]cis-DCE in live microcosms, to as high as 10% 14CO2 from TCE and 20% 14CO2 from cis-DCE. In autoclaved microcosms, significant transformation of [14C]TCE and [14C]cis-DCE also occurred; although some 14CO2 accumulated, the predominant 14C product was soluble and could not be stripped by N2 from an acidic solution (referred to as nonstrippable residue, or NSR). Characterization of the NSR by high-performance liquid and ion chromatography identified glycolate, acetate, and formate as significant components. These results suggest that a combination of abiotic and biotic transformation processes is responsible for attenuation of TCE and cis-DCE in the fractured sandstone aquifer. Tracking the distribution of 14C during the microcosm study was essential for observing these phenomena.     

Demonstration of combined zero-valent iron and electrical resistance heating for in situ trichloroethene remediation

The effectiveness of in situ treatment using zero-valent iron (ZVI) for nonaqueous phase or significant sediment-associated contaminant mass can be limited by relatively low rates of mass transfer to bring contaminants in contact with the reactive media. For a field test in a trichloroethene (TCE) source area, combining moderate-temperature subsurface electrical resistance heating with in situ ZVI treatment was shown to accelerate TCE treatment by a factor of about 4 based on organic daughter products and a factor about 8 based on chloride concentrations. A mass-discharge-based analysis was used to evaluate reaction, dissolution, and volatilization processes at ambient groundwater temperature (~10 °C) and as temperature was increased up to about 50 °C. Increased reaction and contaminant dissolution were observed with increased temperature, but vapor- or aqueous-phase migration of TCE out of the treatment zone was minimal during the test because reactions maintained low aqueous-phase TCE concentrations.     

Multifunctional iron-carbon nanocomposites through an aerosol-based process for the in situ remediation of chlorinated hydrocarbons

Spherical iron-carbon nanocomposites were developed through a facile aerosol-based process with sucrose and iron chloride as starting materials. These composites exhibit multiple functionalities relevant to the in situ remediation of chlorinated hydrocarbons such as trichloroethylene (TCE). The distribution and immobilization of iron nanoparticles on the surface of carbon spheres prevents zerovalent nanoiron aggregation with maintenance of reactivity. The aerosol-based carbon microspheres allow adsorption of TCE, thus removing dissolved TCE rapidly and facilitating reaction by increasing the local concentration of TCE in the vicinity of iron nanoparticles. The strongly adsorptive property of the composites may also prevent release of any toxic chlorinated intermediate products. The composite particles are in the optimal range for transport through groundwater saturated sediments. Furthermore, those iron-carbon composites can be designed at low cost, the process is amenable to scale-up for in situ application, and the materials are intrinsically benign to the environment.     

Multicomponent reactive transport in an in situ zero-valent iron cell

Data collected from a field study of in situ zero-valent iron treatment for TCE were analyzed in the context of coupled transport and reaction processes. The focus of this analysis was to understand the behavior of chemical components, including contaminants, in groundwater transported through the iron cell of a pilot-scale funnel and gate treatment system. A multicomponent reactive transport simulator was used to simultaneously model mobile and nonmobile components undergoing equilibrium and kinetic reactions including TCE degradation, parallel iron dissolution reactions, precipitation of secondary minerals, and complexation reactions. The resulting mechanistic model of coupled processes reproduced solution chemistry behavior observed in the iron cell with a minimum of calibration. These observations included the destruction of TCE and cis-1,2-DCE; increases in pH and hydrocarbons; and decreases in EH, alkalinity, dissolved O2 and CO2, and major ions (i.e., Ca, Mg, Cl, sulfate, nitrate). Mineral precipitation in the iron zone was critical to correctly predicting these behaviors. The dominant precipitation products were ferrous hydroxide, siderite, aragonite, brucite, and iron sulfide. In the first few centimeters of the reactive iron cell, these precipitation products are predicted to account for a 3% increase in mineral volume per year, which could have implications for the longevity of favorable barrier hydraulics and reactivity. The inclusion of transport was key to understanding the interplay between rates of transport and rates of reaction in the field.     

Effect of TCE concentration and dissolved groundwater solutes on NZVI-promoted TCE dechlorination and H2 evolution

Nanoscale zero-valent iron (NZVI) is used to remediate contaminated groundwater plumes and contaminant source zones. The target contaminant concentration and groundwater solutes (NO3-, Cl-, HCO3-, SO4(2-), and HPO4(2-)) should affect the NZVI longevity and reactivity with target contaminants, but these effects are not well understood. This study evaluates the effect of trichloroethylene (TCE) concentration and common dissolved groundwater solutes on the rates of NZVI-promoted TCE dechlorination and H2 evolution in batch reactors. Both model systems and real groundwater are evaluated. The TCE reaction rate constant was unaffected by TCE concentration for [TCE] < or = 0.46 mM and decreased by less than a factor of 2 for further increases in TCE concentration up to water saturation (8.4 mM). For [TCE] > or = 0.46 mM, acetylene formation increased, and the total amount of H2 evolved at the end of the particle reactive lifetime decreased with increasing [TCE], indicating a higher Fe0 utilization efficiency for TCE dechlorination. Common groundwater anions (5mN) had a minor effect on H2 evolution but inhibited TCE reduction up to 7-fold in increasing order of Cl- < SO4(2-) < HCO3- < HPO4(2). This order is consistent with their affinity to form complexes with iron oxide. Nitrate, a NZVI-reducible groundwater solute, present at 0.2 and 1 mN did not affect the rate of TCE reduction but increased acetylene production and decreased H2 evolution. NO3- present at > 3 mM slowed TCE dechlorination due to surface passivation. NO3- present at 5 mM stopped TCE dechlorination and H2 evolution after 3 days. Dissolved solutes accounted for the observed decrease of NZVI reactivity for TCE dechlorination in natural groundwater when the total organic content was small (< 1 mg/L).     

Pd-catalyzed TCE dechlorination in water: effect of [H2](aq) and H2-utilizing competitive solutes on the TCE dechlorination rate and product distribution

The aqueous-phase H2 concentration ([H2](aq)) and the presence of H2-utilizing competitive solutes affect TCE dechlorination efficiency in Pd-based in-well treatment reactors. The effect of [H2](aq) and H2-utilizing competing solutes (cis-DCE, trans-DCE, 1,1-DCE, dissolved oxygen (DO), nitrite, nitrate) on the TCE transformation rate and product distribution were evaluated using 100 mg/L of a powdered Pd-on-Al2O3 catalysts in batch reactors or 1.0 g of a 1.6-mm Pd-on-gamma-Al2O3 catalyst in column reactors. The TCE dechlorination rate constant decreased by 55% from 0.034 +/- 0.006 to 0.015 +/- 0.001 min-1 when the [H2](aq) decreased from 1000 to 100 microM and decreased sharply to 0.0007 +/- 0.0003 min-1 when the [H2](aq) decreased from 100 to 10 microM. Production of reactive chlorinated intermediates and C4-C6 radical coupling products increased with decreasing [H2](aq). At an [H2](aq) of 10 microM (P/Po = 0.01), DCE isomers and vinyl chloride accounted for as much as 9.8% of the TCE transformed at their maximum but disappeared thereafter, and C4-C6 radical coupling products accounted for as much as 18% of TCE transformed. The TCE transformation rate was unaffected by the presence of cis-DCE (202 microM), trans-DCE (89 microM), and 1,1-DCE (91 microM), indicating that these compounds do not compete with TCE for catalyst active sites. DO is twice as reactive as TCE but had no effect on TCE conversion in the column below a concentration of 370 microM (11.8 mg/L), indicating that DO and TCE will not compete for active catalyst sites at typical groundwater DO concentrations. TCE conversion in the column was reduced by as much as a factor of 10 at influent DO levels greater than 450 mM (14.3 mg/L) because the [H2](aq) fell below 100 microM due to H2 utilized in DO conversion. Nitrite reacts 2-5 times slower than TCE and reduced TCE conversion by less than 4% at a concentration of 6630 microM (305 mg/L). Nitrate was not reactive and did not effect TCE conversion at a concentration of 1290 microM (80 mg/L).     

Abiotic degradation of trichloroethylene under thermal remediation conditions

The degradation of trichloroethylene (TCE) to carbon dioxide (CO2) and chloride (Cl-) has been reported to occur during thermal remediation of subsurface environments. The effects of solid-phase composition and oxygen content on the chemical reactivity of TCE were evaluated in sealed ampules that were incubated at 22 and 120 degrees C for periods ranging from 4 to 40 days. For all treatments, no more than 15% of the initial amount of TCE was degraded, resulting in the formation of several non-chlorinated products including Cl-, CO2, carbon monoxide, glycolate, and formate. First-order rate coefficients for TCE disappearance ranged from 1.2 to 6.2 x 10(-3) day(-1) at 120 degrees C and were not dependent upon oxygen content orthe presence of Ottawa sand. However, the rate of TCE disappearance at 120 degrees C increased by more than 1 order-of-magnitude (1.6 to 5.3 x 10(-2) day(-1)), corresponding to a half-life of 13-44 days in ampules containing 1% (wt) goethite and Ottawa sand. These results indicate that the rate of TCE degradation in heated, three-phase systems is relatively insensitive to oxygen content, but may increase substantially in the presence of iron bearing minerals.     

Field evaluation of in situ source reduction of trichloroethylene in groundwater using bioenhanced in-well vapor stripping

Two technologies in combination, cometabolic bioremediation and in-well vapor stripping, were applied to reduce trichloroethylene (TCE) concentrations in groundwater at a contaminant source area without the need to pump contaminated groundwater to the surface for treatment. The vapor-stripping well reduced source TCE concentrations (as high as 6-9 mg/L) by over 95%. Effluent from the well then flowed to two bioremediation wells, where additional reductions of approximately 60% were achieved. TCE removal was extensively monitored (for research and not regulatory purposes) using an automated system that collected samples about every 45 min at 55 locations over an area of approximately 50 x 60 m2. During 4.5 months of system operation, total TCE mass removal was 8.1 kg, 7.1 kg of which resulted from in-well vapor stripping and 1.0 kg from biotreatment. The system reduced the average TCE concentration of about 3000 microg/L in the source-zone groundwater to about 250 microg/L in water leaving the treatment zone, effecting greater than 92% TCE removal. A 6 month rebound study after system operation ceased found TCE concentrations then increased significantly in the treatment zone due to diffusion from the fractured rock below and perhaps other processes, with mass increases of about 1.5 kg in the lower aquifer and 0.3 kg in the upper aquifer.     

Destruction efficiencies and dynamics of reaction fronts associated with the permanganate oxidation of trichloroethylene

Although potassium permanganate (KMnO4) flushing is commonly used to destroy chlorinated solvents in groundwater, many of the problems associated with this treatment scheme have not been examined in detail. We conducted a KMnO4 flushing experiment in a large sand-filled flow tank (L x W x D = 180 cm x 60 cm x 90 cm) to remove TCE emplaced as a DNAPL in a source zone. The study was specifically designed to investigate cleanup progress and problems of pore plugging associated with the dynamics of the solid-phase reaction front (i.e., MnO2) using chemical and optical monitoring techniques. Ambient flow through the source zone formed a plume of dissolved TCE across the flow tank. The volume and concentration of TCE plume diminished with time because of the in situ oxidation of the DNAPL source. The migration velocity of the MnO2 reaction front decreased with time, suggesting that the kinetics of the DNAPL oxidation process became diffusion-controlled because of the pore plugging. A mass balance calculation indicated that only approximately 18% of the total applied KMnO4 (MnO4- = 1250 mg/ L) participated in the oxidation reaction to destroy approximately 41% of emplaced TCE. Evidently, the efficiency of KMnO4 flushing scheme diminished with time due to pore plugging by MnO2 and likely CO2, particularly in the TCE source zone. In addition, the excess KMnO4 used for flushing may cause secondary aquifer contamination. One needs to be concerned about the efficacy of KMnO4 flushing in the field applications. Development of a new approach that can provide both contaminant destruction and plugging/ MnO4- control is required.     

Reductive dechlorination pathways of tetrachloroethylene and trichloroethylene and subsequent transformation of their dechlorination products by mackinawite (FeS) in the presence of metals

Because of frequent co-occurrence of metals with chlorinated organic pollutants, Fe(II), Co(II), Ni(II), and Hg(II) were evaluated for their impact on the dechlorination pathways of PCE and TCE and the subsequent transformation of the initial dechlorination products by FeS. PCE transforms to acetylene via beta-elimination, TCE via hydrogenolysis, and 1,1-DCE via alpha-elimination, while TCE transforms to acetylene via beta-elimination and cis-DCE and 1,1-DCE via hydrogenolysis. Acetylene subsequently transforms in FeS batches, but little transformation of cis-DCE and 1,1-DCE was observed. Branching ratio calculations indicate that the added metals decrease the reductive transformation of PCE and TCE via beta-elimination relative to hydrogenolysis, resulting in a higher production of the toxic DCE byproducts. Nonetheless, acetylene is generally the dominant product. Production of highly water-soluble compound(s) is suspected as a significant source for incomplete mass recoveries. In the transformation of PCE and TCE, the formation of unidentified product(s) is most significant in Co(II)-added FeS batches. Although nearly complete mass recoveries were observed in the other FeS batches, the subsequent transformation of acetylene would lead to the formation of unidentified product(s) over long time periods.     

Efficient degradation of TCE in groundwater using Pd and electro-generated H2 and O2: a shift in pathway from hydrodechlorination to oxidation in the presence of ferrous ions

Degradation of trichloroethylene (TCE) in simulated groundwater by Pd and electro-generated H(2) and O(2) is investigated in the absence and presence of Fe(II). In the absence of Fe(II), hydrodechlorination dominates TCE degradation, with accumulation of H(2)O(2) up to 17 mg/L. Under weak acidity, low concentrations of oxidizing ?OH radicals are detected due to decomposition of H(2)O(2), slightly contributing to TCE degradation via oxidation. In the presence of Fe(II), the degradation efficiency of TCE at 396 μM improves to 94.9% within 80 min. The product distribution proves that the degradation pathway shifts from 79% hydrodechlorination in the absence of Fe(II) to 84% ?OH oxidation in the presence of Fe(II). TCE degradation follows zeroth-order kinetics with rate constants increasing from 2.0 to 4.6 μM/min with increasing initial Fe(II) concentration from 0 to 27.3 mg/L at pH 4. A good correlation between TCE degradation rate constants and ?OH generation rate constants confirms that ?OH is the predominant reactive species for TCE oxidation. Presence of 10 mM Na(2)SO(4), NaCl, NaNO(3), NaHCO(3), K(2)SO(4), CaSO(4), and MgSO(4) does not significantly influence degradation, but sulfite and sulfide greatly enhance and slightly suppress degradation, respectively. A novel Pd-based electrochemical process is proposed for groundwater remediation.     

Trichloroethylene degradation by Ralstonia sp. KN1-10A constitutively expressing phenol hydroxylase: transformation products, NADH limitation, and product toxicity

Ralstonia sp. KN1-10A, which was constructed by inserting the tac promoter upstream of the phenol hydroxylase (PH) gene in the chromosomal DNA of the wild-type strain, Ralstonia sp. KN1, is a useful recombinant strain for eliminating trichloroethylene (TCE) from contaminated sites because it exhibits constitutive TCE oxidation activity. During TCE degradation by Ralstonia sp. KN1-10A, noxious chlorinated compounds, such as dichloroacetic acid, trichloroacetic acid, 2,2,2-trichloroethanol, and chloral, were not detected, and more than 95% of chlorine in TCE was released as chloride ions. Among the possible TCE transformation products, only carbon monoxide was detected, and its conversion percentage was 7 mol%. The addition of formate, which Ralstonia sp. KN1-10A could use as an exogenous electron donor, did not enhance the TCE degradation performance, suggesting that NADH depletion did not limit the degradation. The phenol degradation activity of Ralstonia sp. KN1-10A that previously degraded TCE was not markedly lower than that of cells not exposed to TCE, suggesting that Ralstonia sp. KN1-10A was not susceptible to product toxicity associated with TCE degradation. Furthermore, to clarify the mechanisms underlying TCE degradation by PH from Ralstonia sp. KN1, this enzyme was compared with another enzyme, a hybrid aromatic ring dioxygenase exhibiting a high TCE degradation activity in Escherichia coli and Pseudomonas sp. The initial TCE degradation rate of Ralstonia sp. KN1 (pKTP100), which produced PH, was 1 50 lower than that of Ralstonia sp. KN1 (pKTF200), which produced the hybrid aromatic ring dioxygenase. However, because of its lower product toxicity, the strain producing PH could degrade 2.3 times more TCE than that generated by the strain producing the hybrid aromatic ring dioxygenase.     

Inoculation of a DNAPL source zone to initiate reductive dechlorination of PCE

The ability to inoculate a PCE-NAPL source zone with no prior dechlorinating activity was examined using a near field-scale simulated aquifer. A known mass of PCE was added to establish a source zone, and the groundwater was depleted of oxygen using acetate and lactate prior to culture addition. An active and stable dechlorinating culture was used as an inoculum, and dechlorination activity was observed within 2 weeks following culture transfer. PCE reduction to TCE and cis-DCE was observed initially, and the formation of these compounds was accelerated by the addition of a long-term source of hydrogen (Hydrogen Releasing Compound). cis-DCE was the predominant chlorinated ethene present in the effluent after 225 days of operation, and production of VC and ethene lagged the formation of TCE and cis-OCE. However, dechlorination extent continued to improve over time, and VC eventually became a major product, suggesting that reinoculation was unnecessary. The detection of Dehalococcoides species in the source culture and in the simulated aquifer postinoculation indicated that the metabolic capability to dechlorinate beyond cis-DCE (t = 86 days and t = 245 days) was present. Elevated levels of TCE and cis-DCE were present in the source zone, but neither VC nor ethene were detected in the vicinity of NAPL. The results of this research indicated that adding dechlorinating cultures may be useful in the application of source zone bioremediation but that dechlorination beyond cis-DCE may be limited to regions downgradient of the source zone.