Potential of the TCE-degrading endophyte Pseudomonas putida W619-TCE to improve plant growth and reduce TCE phytotoxicity and evapotranspiration in poplar cuttings

The TCE-degrading poplar endophyte Pseudomonas putida W619-TCE was inoculated in poplar cuttings, exposed to 0, 200 and 400 mg l⁻¹ TCE, that were grown in two different experimental setups. During a short-term experiment, plants were grown hydroponically in half strength Hoagland nutrient solution and exposed to TCE for 3 days. Inoculation with P. putida W619-TCE promoted plant growth, reduced TCE phytotoxicity and reduced the amount of TCE present in the leaves. During a mid-term experiment, plants were grown in potting soil and exposed to TCE for 3 weeks. Here, inoculation with P. putida W619-TCE had a less pronounced positive effect on plant growth and TCE phytotoxicity, but resulted in strongly reduced amounts of TCE in leaves and roots of plants exposed to 400 mg l⁻¹ TCE, accompanied by a lowered evapotranspiration of TCE. Dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA), which are known intermediates of TCE degradation, were not detected.     

A micromodel analysis of factors influencing NAPL removal by surfactant foam flooding

A methodology to study the trichloroethylene (TCE) and dodecane removal in porous media by surfactant foams (SF) was presented by using etched-glass micromodels. The purpose of this work was to systematically evaluate the impact of various physicochemical factors such as gas fraction (GF), surfactant concentration, pore structure and nonaqueous phase liquid (NAPL) types on NAPL removal during SF flooding. The TCE displacement by SF was dependent on the gas fraction of SF. Low GFs (50% and 66%) were more efficient for TCE removal and sweep efficiencies than a high GF (85%). An increase in TCE removal was observed with increasing surfactant concentration at a fixed GF. TCE removal by SF flooding appeared to be dependent more to the value of Capillary number rather than to the concentration of surfactant solution. The effect of the pore heterogeneity was evaluated by employing two different types of micromodels. The Capillary number is an important parameter in the determination of sweep efficiency or gas saturation of SF in a nonhomogeneous porous medium. However, the TCE removal from a nonhomogeneous porous medium may not be associated with sweep efficiency. The initial configuration of residual TCE blobs in a nonhomogeneous porous medium would also be influential in displacing TCE. Sweep efficiencies and pressure responses of two NAPL systems (TCE and dodecane) were monitored to evaluate foam stability when the foam contacts the NAPLs. Stable foam contacting with TCE is implied, while it appears that dodecane cause the SF to collapse. All results indicate that the Capillary number (a ratio of viscous forces to capillary forces) is the most important parameter for TCE removal by SF flooding. Micromodel visualizations of water, surfactant and SF floods were showed and also discussed.     

Mechanistics of trichloroethylene mineralization by the white-rot fungus Trametes versicolor

The white-rot fungus Trametes versicolor degraded trichloroethylene (TCE), a highly oxidized chloroethene, and produced 2,2,2-trichloroethanol and carbon dioxide as the main products of degradation, based on the results obtained using [13C]-TCE as the substrate. For a range of concentrations of TCE between 2 and 20 mg l(-1), 53% of the theoretical maximum chloride expected from complete degradation of TCE was observed. Laccase was shown to be induced by TCE, but did not appear to play a role in TCE degradation. Cytochrome P-450 appears to be involved in TCE degradation, as evidenced by marked inhibition of degradation of TCE in the presence of 1-aminobenzotriazole, a known inhibitor of cytochrome P-450. Our results suggested that chloral (trichloroacetaldehyde) was an intermediate of the TCE degradation pathway. The results indicate that the TCE degradation pathway in T. versicolor appears to be similar to that previously reported in mammals and is mechanistically quite different from bacterial TCE degradation.     

Increased leaf area expansion of hybrid poplar in elevated CO2. From controlled environments to open-top chambers and to FACE.

We examined the response of hybrid poplar to elevated CO2 in contrasting growth environments: controlled environment chamber (CE). open-top chamber (OTC) and poplar free air CO2 enrichment (POPFACE) in order to compare short versus long-term effects and to determine whether generalisations in response are possible for this fast growing tree. Leaf growth, which for poplar is an important determinant of stemwood productivity was followed in all environments, as were the determinants of leaf growth-cell expansion and cell production. Elevated CO2 (550-700 micromol mol(-1), depending on environment) resulted in an increase in final leaf size for Populus trichocarpa x Populus deltoides (Populus x interamericana) and P. deltoides x Populus nigra (Populus x euramericana), irrespective of whether plants were exposed during a short-term CE glasshouse study (90 days), a long-term OTC experiment (3 years) or during the first year of a POPFACE experiment. An exception was observed in the closed canopy POPFACE experiment, where final leaf size remained unaltered by CO2. Increased leaf extension rate was observed in elevated CO2 in all experiments, at some point during leaf development, as determined by leaf length. Again the exception were the POPFACE experiment, where effects were not statistically significant. Leaf production and specific leaf area (SLA) were increased and decreased, respectively, on five out of six occasions, although both were only statistically significant on two occasions and interestingly for SLA never in the FACE experiment. Although both cell expansion and cell production were sensitive to CO2 concentration, effects appeared highly dependent on growth environment and genotype. However, increased leaf cell expansion in elevated CO2 was often associated with changes in the biophysical properties of the cell wall, usually increased cell wall plasticity. This research has shown that enhanced leaf area development was a consistent response to elevated CO2 but that the magnitude of this response is likely to decline, in long-term exposure to elevated CO2. Effects on SLA and leaf production suggest that CE and OTC experiments may not always provide good predictors of the 'qualitative' effects of elevated CO2 in long-term ecosystem experiments.     

The modelling of trichloroethene photodegradation in Brij 35 surfactant by two-stage reaction.

Various clean-up technologies have been developed for the removal and/or destruction of trichloroethene (TCE) in the subsurface. Surfactant-aided soil washing followed by photodegradation could be a promising approach to such a task. The modelling of TCE photodegradation by UV in Brij 35 surfactant micelles is therefore investigated. Two stages of TCE degradation are observed in surfactant Brij 35 systems. A lag phase is observed at the commencement of the degradation, but the duration of the lag phase is significantly reduced as the initial pH increases. As the overall decay of TCE is also found to be faster at higher pH levels, it is suggested that the free radical reaction is dominant at high pH levels, and the formation of lag phases is mainly due to the deficiency of free radicals at lower pH levels. Since the period of the lag phase gradually decreases with the increase of initial pH level, and the two pseudo first-order reaction constants (one for the lag phase and one for the subsequent fast decay) for TCE decay in both stages are also pH dependent, a non-steady-state mathematical model is developed for the prediction of TCE photodegradation in Brij 35 solutions, in which the remaining fraction of TCE (C/C0) in the system can be determined at any instant by using a simple parameter of the initial system pH.     

Reciprocal effects of platinum and lead on the water household of poplar cuttings

In order to study the accumulation rates and effects of platinum as influenced by lead, experiments were performed with poplar cuttings in a growth chamber. The heavy metals were added at a final concentration of 34.8 ppb each to nutrient solutions as PtCl₄ and Pb(NO₃)₂. The variants were 1) control; 2) permanent Pt treatment for 6 weeks; 3) pretreatment with Pt plus subsequent treatment with Pb (three weeks each), and 4) heavy metal application in inverse order to variant 3. The experiments revealed that platinum accumulates in the roots of poplar cuttings to a higher degree than lead. It is translocated from the roots to other plant parts to an extremely low degree. Lead is displaced from the roots by subsequent Pt treatment. Insoluble platinum was found to be associated especially on the cell walls of the rhizodermis and exodermis of the root tips. Accumulation of platinum in the roots leads to a gradual depletion of the plants’ water supply. The disturbance of the water household causes a reduction of the transpirational surface, lowered transpiration rates and enhanced root growth. All these alterations are induced as a means of coping water stress. From the results of this experiment, the conclusion can be drawn that, under the chosen experimental conditions, platinum manifests a higher toxicity than lead in plant roots because of its higher accumulation rates.     

An in situ study of the effect of nitrate on the reduction of trichloroethylene by granular iron

The effect of nitrate on the reduction of TCE by commercial granular iron was investigated in column experiments designed to allow for the in situ monitoring of the iron surface film with Raman spectroscopy. Three column experiments were conducted; one with an influent solution of 100 mg/l nitrate+1.5 mg/l TCE, and two control columns, one saturated directly with 100 mg/l nitrate solution, the other pre-treated with Millipore water prior to the introduction of a 100 mg/l nitrate solution. In the presence of nitrate, TCE adsorbed onto the iron, but there was little TCE reduction to end-products ethene and ethane. The iron used (Connelly, GPM, Chicago) is a product typical of those used in permeable granular iron walls. The material is covered by an air-formed high-temperature oxidation film, consisting of an inner layer of Fe(3)O(4), and an outer, passive layer of Fe(2)O(3). In the control column pre-treated with Millipore water, the passive Fe(2)O(3) layer was removed upon contact with the water in a manner consistent with an autoreduction reaction. In the TCE+nitrate column and the direct nitrate saturation column, nitrate interfered with the removal of the passive layer and maintained conditions such that high valency protective corrosion species, including Fe(2)O(3) and FeOOH, were stable at the iron surface. The lack of TCE reduction is explained by the presence of these species, as they inhibit both mechanisms proposed for TCE reduction by iron, including catalytic hydrogenation, and direct electron transfer.     

Cometabolic degradation kinetics of TCE and phenol by Pseudomonas putida

Modeling of cometabolic kinetics is important for better understanding of degradation reaction and in situ application of bio-remediation. In this study, a model incorporated cell growth and decay, loss of transformation activity, competitive inhibition between growth substrate and non-growth substrate and self-inhibition of non-growth substrate was proposed to simulate the degradation kinetics of phenol and trichloroethylene (TCE) by Pseudomonas putida. All the intrinsic parameters employed in this study were measured independently, and were then used for predicting the batch experimental data. The model predictions conformed well to the observed data at different phenol and TCE concentrations. At low TCE concentrations (<2 mg l(-1)), the models with or without self-inhibition of non-growth substrate both simulated the experimental data well. However, at higher TCE concentrations (>6 mg l(-1)), only the model considering self-inhibition can describe the experimental data, suggesting that a self-inhibition of TCE was present in the system. The proposed model was also employed in predicting the experimental data conducted in a repeated batch reactor, and good agreements were observed between model predictions and experimental data. The results also indicated that the biomass loss in the degradation of TCE below 2 mg l(-1) can be totally recovered in the absence of TCE for the next cycle, and it could be used for the next batch experiment for the degradation of phenol and TCE. However, for higher concentration of TCE (>6 mg l(-1)), the recovery of biomass may not be as good as that at lower TCE concentrations.     

Identification of a novel hydroxylated metabolite of 2,2′,3,5′,6-pentachlorobiphenyl formed in whole poplar plants

Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants consisting of 209 congeners. Oxidation of several PCB congeners to hydroxylated PCBs (OH-PCBs) in whole poplar plants has been reported before. Moreover, 2,2′,3,5′,6-pentachlorobiphenyl (PCB95), as a chiral congener, has been previously shown to be atropselectively taken up and transformed in whole poplar plants. The objective of this study was to determine if PCB95 is atropselectively metabolized to OH-PCBs in whole poplar plants. Two hydroxylated PCB95s were detected by high-performance liquid chromatography-mass spectrometry in the roots of whole poplar plants exposed to racemic PCB95 for 30 days. The major metabolite was confirmed to be 4′-hydroxy-2,2′,3,5′,6-pentachlorobiphenyl (4′-OH-PCB95) by gas chromatography-mass spectrometry (GC-MS) using an authentic reference standard. Enantioselective analysis showed that 4′-OH-PCB95 was formed atropselectively, with the atropisomer eluting second on the Nucleodex β-PM column (E2-4′-OH-PCB95) being slightly more abundant in the roots of whole poplar plants. Therefore, PCB95 can at least be metabolized into 4′-OH-PCB95 and another unknown hydroxylated PCB95 (as a minor metabolite) in whole poplar plants. Both atropisomers of 4′-OH-PCB95 are formed, but E2-4′-OH-PCB95 has greater atropisomeric enrichment in the roots of whole poplar plants. A comparison with mammalian biotransformation studies indicates a distinctively different metabolite profile of OH-PCB95 metabolites in whole poplar plants. Our observations suggest that biotransformation of chiral PCBs to OH-PCBs by plants may represent an important source of enantiomerically enriched OH-PCBs in the environment.     

Real-time reverse transcription PCR analysis of trichloroethylene-regulated toluene dioxygenase expression in Pseudomonas putida F1

Toluene dioxygenase (tod) is a multicomponent enzyme system in Pseudomonas putida F1. Tod can mediate the degradation of Trichloroethylene (TCE), a widespread pollutant. In this study, we try to explore the TCE-regulated tod expression by using real-time qRT-PCR. The minimal culture media were supplemented with glucose, toluene, or a mixture of glucose/toluene respectively as carbon and energy sources. The TCE was injected into each medium after a 12-hour incubation period. The TCE injection severely affected bacterial growth when cultured with toluene or toluene/glucose mixtures. The cell density dropped 61 % for bacteria growing in toluene and 36 % for bacteria in the glucose/toluene mixture after TCE injection, but the TCE treatment had little effect on bacteria supplied with glucose alone. The decrease in cell number was caused by the cytotoxicity of the TCE metabolized by tod. The results from the real-time qRT-PCR revealed that TCE was capable of inducing tod expression in a toluene-dependent manner and that the tod expression level increased 50 times in toluene and 3 times in the toluene/glucose mixture after 6 hours of TCE treatment. Furthermore, validation of the rpoD gene as a reference gene for P. putida F1 was performed in this study, providing a valuable foundation for future studies to use real-time qRT-PCR in the analysis of the P. putida F1 strain.     

硫酸盐还原条件下三氯乙烯的降解研究

三氯乙烯(TCE)是地下水和土壤中主要的有机污染物之一.以受TCE等氯代烃严重污染的地下泥样为材料,采用室内模拟的方法.研究硫酸盐初始质量浓度为100、500、2 500 mg/L时.硫酸盐对TCE降解的影响.结果表明.高浓度硫酸盐更有利于TCE的降解;在硫酸盐还原条件下,通过添加2-溴乙基磺酸钠抑制产甲烷菌.发现产甲烷菌在TCE的降解过程中起到很重要的作用;硫酸盐浓度变化与TCE降解趋势一致.     

Entry and Movement in Vegetation of Lead Derived From Air and Soil Sources

The body of information presented in this paper is directed to individuals concerned with the state of our knowledge on lead uptake and translocation by plants and its subsequent effects. Lead, a non-essential element, is taken up by many plant species primarily via their roots. Large amounts of lead are deposited on plant foliage and most remains as a topical deposit but foliar uptake has been demonstrated. To date it has been assumed that soil lead levels above 1000 ppm are required to cause observable plant effects. Environmental variables, plant age, and species are very important determinants of lead uptake. The few studies done with plant age and speciation, however, provide no clear generalization. Increasing soil lead availability increases plant uptake. Plant uptake decreases with increasing soil phosphorus, organic content, and pH. The lack of observable lead intoxication of native and agricultural plants is surprising in light of evidence that lead concentrations as low as 1 ppm have a profound effect on events associated with photosynthesis and respiration, this has been explained because even though large amounts of lead may be taken up by plant roots they are immobilized by dictyosome vesicles and deposited in the cell wall. It has been suggested that a similar process is operative throughout the plant. The extant data provide no basis for any fear that lead is an imminent dietary hazard to man.     

Evaluation of trichloroethene recovery processes in heterogeneous aquifer cells flushed with biodegradable surfactants

The ability of two biodegradable surfactants, polyoxyethylene (20) sorbitan monooleate (Tween 80) and sodium dihexyl sulfosuccinate (Aerosol MA), to recover a representative dense non-aqueous-phase liquid (DNAPL), trichloroethene (TCE), from heterogeneous porous media was evaluated through a combination of batch and aquifer cell experiments. An aqueous solution containing 3.3% Aerosol MA, 8% 2-propanol and 6 g/l CaCl(2) yielded a weight solubilization ratio (WSR) of 1.21 g TCE/g surfactant, with a corresponding liquid-liquid interfacial tension (IFT) of 0.19 dyn/cm. Flushing of aquifer cells containing a TCE-DNAPL source zone with approximately two pore volumes of the AMA formulation resulted in substantial (>30%) mobilization of TCE-DNAPL. However, a TCE mass recovery of 81% was achieved when the aqueous-phase flow rate was sufficient to displace the mobile TCE-DNAPL toward the effluent well. Aqueous solutions of Tween 80 exhibited a greater capacity to solubilize TCE (WSR=1.74 g TCE/g surfactant) and exerted markedly less reduction in IFT (10.4 dyn/cm). These data contradict an accepted empirical correlation used to estimate IFT values from solubilization capacity, and indicate a unique capacity of T80 to form concentrated TCE emulsions. Flushing of aquifer cells with less than 2.5 pore volumes of a 4% T80 solution achieved TCE mass recoveries ranging from 66 to 85%, with only slight TCE-DNAPL mobilization (<5%) occurring when the total trapping number exceeded 2 x 10(-5). These findings demonstrate the ability of Tween 80 and Aerosol MA solutions to efficiently recover TCE from a heterogeneous DNAPL source zone, and the utility of the total trapping number as a design parameter for a priori prediction of DNAPL mobilization and bank angle formation when flushing with low-IFT solutions. Given their potential to stimulate microbial reductive dechlorination at low concentrations, these surfactants are well-suited for remedial action plans that couple aggressive mass removal followed by enhanced bioremediation to treat chlorinated solvent source zones.     

The rate improvement and modeling of trichloroethene photodegradation by acetone sensitizer in surfactant solution

The photodecay of trichloroethene (TCE) in surfactant solution by the help of photosensitizer (acetone, ACE) was investigated and modeled. Apart from the direct photodegradation, photosensitization is presumed to be one of the major mechanisms contributing to overall decay. Quantum yields of TCE photodecay in solution with surfactant Brij 35 and optimal ACE dosage are about 25 times higher than in Brij 35 alone. However, with an excess ACE dosage, ACE will act as a light barrier and attenuate the light intensity available for TCE photodegradation. TCE photodegradation follows a two-stage kinetics, in which a lag phase is followed by a fast decay. Mathematical models were developed for the prediction of the two-stage photodegradation, in which the remaining fraction of TCE (C/C0) in the system becomes predictable.     

Carbon isotope fractionation during reductive dechlorination of TCE in batch experiments with iron samples from reactive barriers

Reductive dechlorination of trichloroethene (TCE) by zero-valent iron produces a systematic enrichment of 13C in the remaining substrate that can be described using a Rayleigh model. In this study, fractionation factors for TCE dechlorination with iron samples from two permeable reactive barriers (PRBs) were established in batch experiments. Samples included original unused iron as well as material from a barrier in Belfast after almost 4 years of operation. Despite the variety of samples, carbon isotope fractionations of TCE were remarkably similar and seemed to be independent of iron origin, reaction rate, and formation of precipitates on the iron surfaces. The average enrichment factor for all experiments was -10.1 per thousand (+/- 0.4 per thousand). These results indicate that the enrichment factor provides a powerful tool to monitor the reaction progress, and thus the performance, of an iron-reactive barrier over time. The strong fractionation observed may also serve as a tool to distinguish between insufficient residence time in the wall and a possible bypassing of the wall by the plume, which should result in an unchanged isotopic signature of the TCE. Although further work is necessary to apply this stable isotope method in the field, it has potential to serve as a unique monitoring tool for PRBs based on zero-valent iron.     

Clonal differences in survival capacity, copper and zinc accumulation, and correlation with leaf polyamine levels in poplar: A large-scale field trial on heavily polluted soil

Three ex situ collections of poplar clones from natural populations of Populus alba and P. nigra growing in northern Italy were assessed for their genetic dissimilarity (GD) by means of amplified fragment length polymorphism (AFLP). The high GD evidenced within populations was exploited for screening 168 clones in a field trial on heavy metal-polluted soil. After one growth season, clonal differences in plant survival and growth were observed. On the basis of performance, six clones were singled out, and used to evaluate copper and zinc accumulation in different organs. Clonal differences in metal concentrations were most evident for leaves and stems; one clone of P. alba (AL35) had a distinctly higher concentration of both metals in the roots. Leaf polyamine (putrescine, spermidine, spermine) profiles correlated with tissue metal concentrations, depending on the clone, plant organ and metal. In particular, the high metal-accumulating clone AL35 exhibited a dramatically higher concentration of free and conjugated putrescine. Overall, the results indicate that, given the high GD of Populus even within populations, it is possible to identify genotypes best suited for soil clean-up, and useful also for investigating physiological markers associated with high metal accumulation/tolerance     

Evaluation of Vegetation Near Coal-burning Power Plants in Southwestern Pennsylvania. II. Ozone Injury on Foliage of Hybrid Poplar

The objective of this study was to determine if the incidence or severity of foliar injury induced by regional, ambient ozone was influenced by local emissions from a complex of coal-burning power plants in southwestern Pennsylvania. Plantings of an ozonesensitive hybrid poplar clone {Populus maximowizii x trichocarpa, clone NE 388) were established in 1972 at various distances and directions from the power plants. Foliar injury caused by ambient ozone was evaluated annually from 1973 to 1990 in early to mid- August. Data are presented for the 12-year period, 1979 to 1990 inclusive, for which the most complete data sets were available. Injury from ambient ozone varied spatially and temporally, but with little relationship to power plant location. There was an apparent negative relationship between emission trends and ozone-induced symptoms, but only for one power plant. The correlation between annual mean levels of ozone-induced stipple and frequency of days (per year) with a 1-hr ozone maximum exceeding 0.04 ppm was weak, but significant. Ozone-induced bifacial necrosis was not observed on the foliage of the hybrid poplar during the drought year of 1988 in spite of record high levels of ozone; however, ozoneinduced stipple was observed.     

Laboratory column studies for evaluating a barrier system for providing oxygen and substrate for TCE biodegradation

The industrial solvent trichloroethylene (TCE) is among the most ubiquitous chlorinated compounds found in groundwater contamination. The objective of this study was to develop a biobarrier system containing oxygen-organic releasing material to enhance the aerobic cometabolism of TCE in situ. The oxygen-organic material, which contains calcium peroxide and peat, is able to release oxygen and primary substrates continuously upon contact with water. Batch experiments were conducted to design and identify the components of the oxygen-organic releasing material, and evaluate the oxygen and organic substrate (presented as COD equivalent) release from the designed oxygen-organic material. The observed oxygen and chemical oxygen demand (COD) release rates were approximately 0.0246 and 0.052 mg/d/g of material, respectively. A laboratory-scale column experiment was then conducted to evaluate the feasibility of this proposed system for the bioremediation of TCE-contaminated groundwater. This system was performed using a series of continuous-flow glass columns including a soil column, an oxygen-organic material column, followed by two consecutive soil columns. Aerobic acclimated sludges were inoculated in all three soil columns to provide microbial consortia for TCE biodegradation. Simulated TCE-contaminated groundwater with a flow rate of 0.25 l/day was pumped into this system. Effluent samples from each column were analyzed for TCE and other indicating parameters (e.g., pH, dissolved oxygen). Results show that the decreases in TCE concentrations were observed over a 4-month operating period. Up to 99% of TCE removal efficiency was obtained in this passive system. Results indicate that the continuously released oxygen and organic substrates from the oxygen-organic materials enhanced TCE biotransformation. Thus, the biobarrier treatment scheme has the potential to be developed into an environmentally and economically acceptable remediation technology.     

Self-inhibition can limit biologically enhanced TCE dissolution from a TCE DNAPL

Biodegradation of trichloroethene (TCE) near a Dense Non Aqueous Phase Liquid (DNAPL) can enhance the dissolution rate of the DNAPL by increasing the concentration gradient at the DNAPL-water interface. Two-dimensional flow-through sand boxes containing a TCE DNAPL and inoculated with a TCE dechlorinating consortium were set up to measure this bio-enhanced dissolution under anaerobic conditions. The total mass of TCE and daughter products in the effluent of the biotic boxes was 3-6 fold larger than in the effluent of the abiotic box. However, the mass of daughter products only accounted for 19-55% of the total mass of chlorinated compounds in the effluent, suggesting that bio-enhanced dissolution factors were maximally 1.3-2.2. The enhanced dissolution most likely primarily resulted from variable DNAPL distribution rather than biodegradation. Specific dechlorination rates previously determined in a stirred liquid medium were used in a reactive transport model to identify the rate limiting factors. The model adequately simulated the overall TCE degradation when predicted resident microbial numbers approached observed values and indicated an enhancement factor for TCE dissolution of 1.01. The model shows that dechlorination of TCE in the 2D box was limited due to the short residence time and the self-inhibition of the TCE degradation. A parameter sensitivity analysis predicts that the bio-enhanced dissolution factor for this TCE source zone can only exceed a value of 2 if the TCE self-inhibition is drastically reduced (when a TCE tolerant dehalogenating community is present) or if the DNAPL is located in a low-permeable layer with a small Darcy velocity.