Biodegradation of creosote and pentachlorophenol in contaminated groundwater: chemical and biological assessment.

Shake flask studies examined the rate and extent of biodegradation of pentachlorophenol (PCP) and 42 components of coal-tar creosote present in contaminated groundwater recovered from the American Creosote Works Superfund site, Pensacola, Fla. The ability of indigenous soil microorganisms to remove these contaminants from aqueous solutions was determined by gas chromatographic analysis of organic extracts of biotreated groundwater. Changes in potential environmental and human health hazards associated with the biodegradation of this material were determined at intervals by Microtox assays and fish toxicity and teratogenicity tests. After 14 days of incubation at 30 degrees C, indigenous microorganisms effectively removed 100, 99, 94, 88, and 87% of measured phenolic and lower-molecular-weight polycyclic aromatic hydrocarbons (PAHs) and S-heterocyclic, N-heterocyclic, and O-heterocyclic constituents of creosote, respectively. However, only 53% of the higher-molecular-weight PAHs were degraded; PCP was not removed. Despite the removal of a majority of the organic contaminants through biotreatment, only a slight decrease in the toxicity and teratogenicity of biotreated groundwater was observed. Data suggest that toxicity and teratogenicity are associated with compounds difficult to treat biologically and that one may not necessarily rely on indigenous microorganisms to effectively remove these compounds in a reasonable time span; to this end, alternative or supplemental approaches may be necessary. Similar measures of the toxicity and teratogenicity of treated material may offer a simple, yet important, guide to bioremediation effectiveness.     

Monitoring of Desulfitobacterium frappieri PCP-1 in pentachlorophenol-degrading anaerobic soil slurry reactors

Anaerobic biodegradation of pentachlorophenol (PCP) was studied in rotative bioreactors containing 200 g of PCP-contaminated soil and 250 ml of liquid medium. Reactors were bioaugmented with cells of Desulfitobacterium frappieri strain PCP-1, a bacterium able to dehalogenate PCP to 3-chlorophenol. Cells of strain PCP-1 were detected by quantitative PCR for at least 21 days in reactors containing 500 mg of PCP per kg of soil but disappeared after 21 days in reactors with 750 mg of PCP per kg of soil. Generally, PCP was completely removed in less than 9 days in soils contaminated with 189 mg of PCP per kg of soil. Sorption of PCP to soil organic matter reduced its toxicity and enhanced the survival of strain PCP-1. In some non-inoculated reactors, the indigenous microorganisms of some soils were also able to degrade PCP. These results suggest that anaerobic dechlorination of PCP in soils by indigenous PCP-degrading bacteria, or after augmentation with D. frappieri PCP-1, should be possible in situ and ex situ when the conditions are favourable for the survival of the degrading microorganisms.     

Adsorption of polycyclic aromatic hydrocarbons (PAHs) by soil particles: influence on biodegradability and biotoxicity

Summary Polycyclic aromatic hydrocarbon (PAH) biodegradation was investigated in contaminated soils from two different industrial sites under simulated land treatment conditions. Soil samples from a former impregnation plant (soil A) showed high degradation rates of PAHs by the autochthonous microorganisms, whereas PAHs in material of a closed-down coking plant (soil B) were not degraded even after inoculation with bacteria known to effectively degrade PAHs. As rapid PAH biodegradation in soil B was observed after PAHs were extracted and restored into the extracted soil material, the kind of PAH binding in soil B appears to completely prevent biodegradation. Sorption of PAHs onto extracted material of soil B follows a two-phase process (fast and slow); the latter is discussed in terms of migration of PAHs into soil organic matter, representing less accessible sites within the soil matrix. Such sorbed PAHs are suggested to be non-bioavailable and thus non-biodegradable. By eluting soil B with water, no biotoxicity, assayed as inhibition of bioluminescence, was detected in the aqueous phase. When treating soil A analogously, a distinct toxicity was observed, which was reduced relative to the amount of activated carbon added to the soil material. The data suggest that sorption of organic pollutants onto soil organic matter significantly affects biodegradability as well as biotoxicity.     

Potential use of organomineral complex (OMC) for bioremediation of pentachlorophenol (PCP) in soil

Adsorption/desorption characteristics for the organic pollutant pentachlorophenol (PCP) were determined for the organomineral complex (OMC) prepared in the laboratory with clay mineral (zeolite–clinoptilolite) and organic matter (humic acids), both natural products with excellent sorption properties. Sorption experiments were carried out in three characterized soil samples, Calcaro-haplic Chernozem, Gleyic Fluvisol, and Arenic Regosol. The results of this study indicate that OMC has better retention abilities than the clay minerals alone. Higher amounts of humic acids (HAs), bound to zeolite, enhance its potential to adsorb and retain PCP. An OMC containing approx. 5 mg HA g⁻¹ of zeolite possessed the best retention ability for PCP and presented an optimal economic solution from the preparation point of view. Then biodegradation of PCP was studied in the same types of sterilized soils bioaugmented with the bacterial isolate Comamonas testosteroni CCM 7350, with and without the addition of OMC. The immobilization effect of OMC in relation to PCP depends on the concentration of PCP and the content of organic carbon in the soil. The activity of the microorganisms and the effect of acid rain led to the gradual release and biodegradation of the irreversibly bound PCP, without any initial toxic effect on indigenous or bioaugmented microorganisms. OMC appeared to be a good adsorbant for PCP, with potential application in remediation technology. Fast and effective adsorption and low desorption may serve as a pretreatment step for bioremediation technology for reducing PCP content in soil and thus for reducing its potential toxicity, reducing bioavailability, and in this way facilitating biodegradation.     

Degradation of xenobiotic compounds in situ: Capabilities and limits

Abstract: Exploiting microorganisms for remediation of waste sites is a promising alternative to groundwater pumping and above ground treatment. The objective of in situ bioremediation is to stimulate the growth of indigenous or introduced microorganisms in regions of subsurface contamination, and thus to provide direct contact between microorganisms and the dissolved and sorbed contaminants for biotransformation. Subsurface microorganisms detected at a former manufactured gas plant site contaminated with coal tars mineralized significant amounts of naphthalene (8–43%) and phenanthrene (3–31%) in sediment-water microcosms incubated for 4 weeks under aerobic conditions. Evidence was obtained for naphthalene mineralization (8–13%) in the absence of oxygen in field samples. These data suggest that biodegradation of these compounds is occurring at the site, and the prospects are good for enhancing this biodegradation. Additional batch studies demonstrated that sorption of naphthalene onto aquifer materials reduced the extent and rate of biodegradation, indicating that desorption rate was controlling the biodegradation performance.     

Biodesulfurization of water-soluble coal-derived material by Rhodococcus rhodochrous IGTS8

Rhodococcus rhodochrous IGTS8 was previously isolated because of its ability to use coal as its sole source of sulfur for growth. Subsequent growth studies have revealed that IGTS8 is capable of using a variety of organosulfur compounds as sources of sulfur but not carbon. In this article, the ability of IGTS8 to selectively remove organic sulfur from water-soluble coal-derived material is investigated. The microbial removal of organic sulfur from coal requires microorganisms capable of cleaving carbon-sulfur bonds and the accessibility of these bonds to microorganisms. The use of water-soluble coal-derived material effectively overcomes the problem of accessibility and allows the ability of microorganisms to cleave carbon-sulfur bonds present in coal-derived material to be assessed directly. Three coals, two coal solubilization procedures, and two methods of biodesulfurization were examined. The results of these experiments reveal that the microbial removal of significant amounts of organic sulfur from water-soluble coal-derived material with treatment times as brief as 24 h is possible. Moreover, the carbon content and calorific value of biotreated products are largely unaffected. Biotreatment does result, however, in an increased hydrogen and nitrogen content and a decreased oxygen content of the coal-derived material. The aqueous supernatant obtained from biodesulfurization experiments does not contain sulfate, sulfite, or other forms of soluble sulfur at increased concentrations in comparison with control samples. Sulfur removed from water-soluble coal-derived material appears to be incorporated into biomass. (c) 1992 John Wiley & Sons, Inc.     

Assessment of Factors Governing Biodegradability of PAHs in Three Soils Aged Under Field Conditions

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants of great environmental concern due to their toxic, mutagenic and carcinogenic properties. This study correlates soil characteristics (i.e. soil organic matter, particle- and pore-size distribution) with extractability and toxicity data (LUMIStox, Ostracod) to investigate factors that govern biodegradability of PAHs in three historically contaminated soils. Desorption of PAHs occurred most readily from soil TA1 (82%), followed by soil AS3.7 (69%) and soil WG2 (20%). This is in line with toxicity data, as the soil in which the greatest contaminant desorption (SFE) was observed exhibited the highest toxicity (TA1). Of the three soils, pronounced biodegradation of 2-4-ring, and slight biodegradation of 5-ring PAHs was observed only in AS3.7, while no decrease of PAHs was reported for soils WG2 and TA1 during the degradation experiment. Strong sorption reduced pollutant bioavailability in WG2 and hence hampered biodegradation. By contrast, pollutant sorption was weak in TA1 and microbial activity was most likely inhibited due to high toxicity in this soil. Based on our results we conclude that biodegradation of PAHs in soils is determined by a number of phenomena with complex interactions between them. Consideration of a single factor will be misleading and may result in false prediction of the biodegradation potential.     

Effect of modified Fenton's reaction on microbial activity and removal of PAHs in creosote oil contaminated soil

This study describes the removal of polycyclic aromatic hydrocarbons (PAHs) from creosote oil contaminated soil by modified Fenton's reaction in laboratory-scale column experiments and subsequent aerobic biodegradation of PAHs by indigenous bacteria during incubation of the soil. The effect of hydrogen peroxide addition for 4 and 10 days and saturation of soil with H(2)O(2) on was studied. In both experiments the H(2)O(2) dosage was 0.4 g H(2)O(2)/g soil. In completely H(2)O(2)-saturated soil the removal of PAHs (44% within 4 days) by modified Fenton reaction was uniform over the entire soil column. In non-uniformly saturated soil, PAH removal was higher in completely saturated soil (52% in 10 days) compared to partially saturated soil, with only 25% in 10 days. The effect of the modified Fenton's reaction on the microbial activity in the soil was assessed based on toxicity tests towards Vibrio fischeri, enumeration of viable and dead cells, microbial extracellular enzyme activity, and oxygen consumption and carbon dioxide production during soil incubation. During the laboratory-scale column experiments, the toxicity of column leachate towards Vibrio fischeri increased as a result of the modified Fenton's reaction. The activities of the microbial extracellular enzymes acetate- and acidic phosphomono-esterase were lower in the incubated modified Fenton's treated soil compared to extracellular enzyme activities in untreated soil. Abundance of viable cells was lower in incubated modified Fenton treated soil than in untreated soil. Incubation of soil in serum bottles at 20 degrees C resulted in consumption of oxygen and formation of carbon dioxide, indicating aerobic biodegradation of organic compounds. In untreated soil 20-30% of the PAHs were biodegraded during 2 months of incubation. Incubation of chemically treated soil slightly increased PAH-removal compared to PAH-removal in untreated soil.     

Polycyclic aromatic hydrocarbons: environmental pollution and bioremediation

Polycyclic aromatic hydrocarbons (PAHs) are widely distributed and relocated in the environment as a result of the incomplete combustion of organic matter. Many PAHs and their epoxides are highly toxic, mutagenic and/or carcinogenic to microorganisms as well as to higher systems including humans. Although various physicochemical methods have been used to remove these compounds from our environment, they have many limitations. Xenobiotic-degrading microorganisms have tremendous potential for bioremediation but new modifications are required to make such microorganisms effective and efficient in removing these compounds, which were once thought to be recalcitrant. Metabolic engineering might help to improve the efficiency of degradation of toxic compounds by microorganisms. However, efficiency of naturally occurring microorganisms for field bioremediation could be significantly improved by optimizing certain factors such as bioavailability, adsorption and mass transfer. Chemotaxis could also have an important role in enhancing biodegradation of pollutants. Here, we discuss the problems of PAH pollution and PAH degradation, and relevant bioremediation efforts.     

Bioremediation of creosote-contaminated soil: a pilot-scale landfarming evaluation

A pilot-scale landfarming investigation of the effects of biostimulation and bioaugmentation on a creosote-contaminated (258.3 g kg^–1) mispah form (FAO: lithosol) soil, with a view to developing a cost-effective bioremediation methodology for creosote-contaminated soils was conducted in nine duplicate reactors, including two controls (Treatments 1 and 2). Treatments 3–9 were watered and aerated daily and Treatment 4–9 were monthly amended with mono-ammonium phosphate. Treatment 5–9 received further amendments as follows: Treatment 5, hydrogen peroxide; Treatment 6, indigenous microbial biosupplement; Treatment 7, sewage sludge; Treatment 8, cow manure; Treatment 9, poultry manure. Residual concentrations of creosote ranged between 29 and 215 g kg^–1 after sixteen weeks. The phenolics and the 2- and 3-ringed polyaromatic hydrocarbons (PAHs) were removed below detectable levels or to very low levels. The 4- and 5-ringed PAHs were removed by between 68 and 83%. Indigenous microbial biosupplement and sewage sludge were the most effective in creosote removal. Hydrogen peroxide did not significantly enhance microbial population and creosote removal. There was no significant difference between the results obtained from the treatments amended with organic manures. However, there was a significant difference between the effects of the organic manures and the indigenous microbial biosupplement. Results from this study suggests that a combination of the two treatment techniques (biostimulation and bioaugmentation) would be a better approach to treating soil contaminated with very high concentrations of creosote.     

Salt marsh plant-microorganism interaction in the presence of mixed contamination

In this study we investigated under laboratory conditions, the combined effect of the presence of two contaminants (Cu and PAHs) and plants (Halimione portulacoides) on a salt marsh microbial community, in terms of genetic structure, abundance and capacity to remove those contaminants. Plants changed the microbial community structure (evaluated by ARISA), but only in the treatments without PAHs. Also, in the presence of plants, Cu displayed lower toxicity (ToxScreen test) than in its absence. Nevertheless, the presence of plants interfered with the degradation (decreasing it up to 45%) of higher molecular weight PAHs by sediment microorganisms. This was in agreement with the lower microbial numbers (quantified by DAPI) observed in the presence of the plants, pointing for a competition between plants and microorganisms for nutrients. These results are new for salt marshes, and highlight the need for fertilization in order to obtain optimal effects of rhizoremediation for this type of compounds.     

Aerobic and Anaerobic Biodegradation of Aged Pentachlorophenol by Indigenous Microorganisms

Pentachlorophenol (PCP) use as a general biocide, particularly for treating wood, has led to widespread environmental contamination. Biodegradation has emerged as the main mechanism for PCP degradation in soil and groundwater and a key strategy for remediation. Examining the microbial biodegrading potential for PCP at a contaminated site is crucial in determining its fate. Hundreds of studies have been published on PCP microbial degradation, but few have described the biodegradation of PCP that has been in contact with soils for many years. The bioavailability of “aged” hydrophobic organics is a significant concern. PCP- and 2,3,4,6-tetrachlorophenol (2,3,4,6-TeCP)-contaminated soil samples from several depths at a former wood treatment site were placed under varying conditions in the laboratory to determine the anaerobic and aerobic potential for biodegradation of chlorophenols at the site. PCP biodegradation occurred in both anaerobic and aerobic soil samples. Rapid aerobic degradation occurred in samples spiked with 2- and 4-chlorophenol, but not with 3-chlorophenol. Reductive dechlorination of PCP in anaerobic samples resulted in the accumulation of 3-chlorophenol. In most anaerobic replicates, 3-chlorophenol was degraded with the appearance of detectable, but not quantifiable amounts of phenol. These results indicate excellent potential for remediation at the site using the indigenous microorganisms under both aerobic and anaerobic conditions. However, a fraction of the PCP was unavailable for degradation.     

Stable isotope probing identifies anthracene degraders under methanogenic conditions

Polycyclic aromatic hydrocarbons (PAHs) are common contaminants in groundwater. The remediation of PAH-contaminated groundwater often involves anaerobic biodegradation. The knowledge about the microorganisms responsible for PAH degradation in anaerobic subsurface environment is still lacking. DNA-based stable isotope probing (SIP) was applied to discover the microorganisms responsible for anaerobic anthracene degradation within microcosms inoculated with aquifer sediment from landfill leachate-contaminated site. Three phylotypes were identified as the degraders, all falling within the phylum Proteobacteria. Two anthracene degraders were classified within the genera Methylibium and Legionella, while another one was an unclassified Rhizobiales species. They all were first linked to PAH degradation. These findings also provide an illustration of the utility of SIP to discover the roles of uncultured microorganisms in PAH-degrading processes.     

Effect of Modified Fenton’s Reaction on Microbial Activity and Removal of PAHs in Creosote Oil Contaminated Soil

This study describes the removal of polycyclic aromatic hydrocarbons (PAHs) from creosote oil contaminated soil by modified Fenton’s reaction in laboratory-scale column experiments and subsequent aerobic biodegradation of PAHs by indigenous bacteria during incubation of the soil. The effect of hydrogen peroxide addition for 4 and 10 days and saturation of soil with H₂O₂ on was studied. In both experiments the H₂O₂ dosage was 0.4 g H₂O₂/g soil. In completely H₂O₂−saturated soil the removal of PAHs (44% within 4 days) by modified Fenton reaction was uniform over the entire soil column. In non-uniformly saturated soil, PAH removal was higher in completely saturated soil (52% in 10 days) compared to partially saturated soil, with only 25% in 10 days. The effect of the modified Fenton’s reaction on the microbial activity in the soil was assessed based on toxicity tests towards Vibrio fischeri, enumeration of viable and dead cells, microbial extracellular enzyme activity, and oxygen consumption and carbon dioxide production during soil incubation. During the laboratory-scale column experiments, the toxicity of column leachate towards Vibrio fischeri increased as a result of the modified Fenton’s reaction. The activities of the microbial extracellular enzymes acetate- and acidic phosphomono-esterase were lower in the incubated modified Fenton’s treated soil compared to extracellular enzyme activities in untreated soil. Abundance of viable cells was lower in incubated modified Fenton treated soil than in untreated soil. Incubation of soil in serum bottles at 20 °C resulted in consumption of oxygen and formation of carbon dioxide, indicating aerobic biodegradation of organic compounds. In untreated soil 20–30% of the PAHs were biodegraded during 2 months of incubation. Incubation of chemically treated soil slightly increased PAH-removal compared to PAH-removal in untreated soil.     

In vitro toxicity evaluation of organic extract of landfill soil and its detoxification by indigenous pyrene-degrading Bacillus sp. ISTPY1

Organic contaminants present in okhla landfill soil belong to the group polyaromatic hydrocarbons, pharmaceutical compounds, steroidal compounds, personal care products and their derivatives. An indigenous pyrene-degrading Bacillus sp. ISTPY1 was used to treat the Okhla landfill soil. GC–MS analysis of the organic extract before and after biodegradation with Bacillus sp. ISTPY1 showed the elimination of various polyaromatic hydrocarbons and other persistent aromatic compounds. Toxicity study was done on human hepato-carcinoma cell line HepG2 before and after treatment. The bacterium treated sample initially showed reduction in toxicity till 48 h. This was increased after 120 h due to formation of quinones intermediate and further decreased after 360 h. The LC50 value (MTT assay) also showed the same pattern. The reduction in Olive Tail Moment was observed after 360 h treatment. Result of the study indicated biodegradation and detoxification of major contaminants of Okhla landfill by Bacillus sp. ISTPY1.     

Maintenance and stability of introduced genotypes in groundwater aquifer material

Three indigenous groundwater bacterial strains and Pseudomonas putida harboring plasmids TOL (pWWO) and RK2 were introduced into experimentally contaminated groundwater aquifer microcosms. Maintenance of the introduced genotypes was measured over time by colony hybridization with gene probes of various specificity. On the basis of the results of colony hybridization quantitation of the introduced organisms and genes, all introduced genotypes were stably maintained at approximately 10(5) positive hybrid colonies g-1 of aquifer microcosm material throughout an 8-week incubation period. Concomitant removal of the environmental contaminants, viz., toluene, chlorobenzene, and styrene, in both natural (uninoculated) and inoculated aquifer microcosms was also demonstrated. The results indicate that introduced catabolic plasmids, as well as indigenous organisms, can be stably maintained in groundwater aquifer material without specific selective pressure for the introduced genotypes. These results have positive implications for in situ treatment and biodegradation in contaminated aerobic groundwater aquifers.     

Maintenance and stability of introduced genotypes in groundwater aquifer material.

Three indigenous groundwater bacterial strains and Pseudomonas putida harboring plasmids TOL (pWWO) and RK2 were introduced into experimentally contaminated groundwater aquifer microcosms. Maintenance of the introduced genotypes was measured over time by colony hybridization with gene probes of various specificity. On the basis of the results of colony hybridization quantitation of the introduced organisms and genes, all introduced genotypes were stably maintained at approximately 10(5) positive hybrid colonies g-1 of aquifer microcosm material throughout an 8-week incubation period. Concomitant removal of the environmental contaminants, viz., toluene, chlorobenzene, and styrene, in both natural (uninoculated) and inoculated aquifer microcosms was also demonstrated. The results indicate that introduced catabolic plasmids, as well as indigenous organisms, can be stably maintained in groundwater aquifer material without specific selective pressure for the introduced genotypes. These results have positive implications for in situ treatment and biodegradation in contaminated aerobic groundwater aquifers.     

Bioremediation of PAHs-contaminated soil through composting: Influence of bioaugmentation and biostimulation on contaminant biodegradation

The degradation of several polycyclic aromatic hydrocarbons (PAHs) in soil through composting was investigated. The selected PAHs included: fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo(a)anthracene, and chrysene, with concentrations simulating a real creosote sample. The degradation of PAHs (initial concentration 1 g of total PAHs kg⁻¹ dry soil) was assessed applying bioaugmentation with the white-rot fungi Trametes versicolor and biostimulation using compost of the source-selected organic fraction of municipal solid waste (OFMSW) and rabbit food as organic co-substrates. The process performance during 30 days of incubation was evaluated through different analyses including: dynamic respiration index (DRI), cumulative oxygen consumption during 5 days (AT₅), enzymatic activity, and fungal biomass. These analyses demonstrated that the introduced T. versicolor did not significantly enhance the degradation of PAHs. However, biostimulation was able to improve the PAHs degradation: 89% of the total PAHs were degraded by the end of the composting period (30 days) compared to the only 29.5% that was achieved by the soil indigenous microorganisms without any co-substrate (control, not amended). Indeed, the results showed that stable compost from the OFMSW has a greater potential to enhance the degradation of PAHs compared to non-stable co-substrates such as rabbit food.     

Dual substrate biodegradation of a nonionic surfactant and pentachlorophenol by Sphingomonas chlorophenolica RA2

The simultaneous biodegradation of the nonionic surfactant Tween 20 (Tw20) and pentachlorophenol (PCP) by Sphingomonas chlorophenolica sp. Strain RA2 (RA2) was measured. As a sole substrate, Tw20 biodegradation was best described by the Contois kinetic model. During concurrent biodegradation of Tw20 and PCP, the biodegradation rates of Tw20 were not significantly affected by 50 or 100 mg/L PCP, but were significantly inhibited by 500 mg/L PCP. Decreases in cell yield in the presence of PCP suggest that PCP was acting as an uncoupler. Cultures were pre-grown on PCP or Tw20 before degradation of PCP to evaluate enzyme induction effects, and long lags before PCP biodegradation after growth on Tw20 occurred. Although biokinetic models could accurately describe some of the data sets of RA2 growth and Tw20 and PCP degradation, finding a single set of kinetic parameters that predicted all dual substrate tests was not achieved. The complicating factors to modeling PCP and Tw20 interactions are described and may be more widely applicable to the biodegradation of toxic organic compounds in the presence of a biodegradable surfactant.     

Biodegradation of Trichloroethylene and Dichloromethane in Contaminated Soil and Groundwater

Soil column and serum bottle microcosm experiments were conducted to investigate the potential for in situ anaerobic bioremediation of trichloroethy lene (TCE) and dichloromethane (DCM) at the Pinellas site near Largo, Florida. Soil columns with continuous groundwater recycle were used to evaluate treatment with complex nutrients (casamino acids, methanol, lactate, sulfate); benzoate and sulfate; and methanol. The complex nutrients drove microbial dechlorination of TCE to ethene, whereas the benzoate/sulfate and methanol supported microbial dechlorination of TCE only to cis-1 ,2-dichloroethylene (cDCE). Microbial sulfate depletion in the benzoate/sulfate column allowed further dechlorination of cDCE to vinyl chloride. Serum bottle microcosms were used to investigate TCE dechlorination and DCM biodegradation in Pinellas soil slurries bioaugmented with liquid from the soil columns possessing TCE-dechlorinating activity and DCM biodegradation by indigenous microorganisms. Bioaugmented soil microcosms showed immediate TCE dechlorination in the microcosms with methanol or complex nutrients, but no dechlorination in the benzoate/sulfate microcosm. DCM biodegradation by indigenous microorganisms occurred in soil microcosms amended with either benzoate/sulfate or methanol, but not with complex nutrients. Bioaugmentation stimulated DCM biodegradation in both complex nutrient and methanol-amended microcosms, but appeared to inhibit DCM biodegradation in benzoate/sulfate-amended microcosms. TCE dechlorination occurred before DCM biodegradation in bioaugmented microcosms when both compounds were present.