Static and push-pull methods using radon-222 to characterize dense nonaqueous phase liquid saturations Davis BM, Istok JD, Semprini L GROUND WATER 41 (4): 470-481 JUL-AUG 2003 Abstract: Naturally occurring radon in ground water can potentially be used as an in situ partitioning tracer to characterize dense nonaqueous phase liquid (DNAPL) saturations. The static method involves comparing radon concentrations in water samples from DNAPL-contaminated and noncontaminated portions of an aquifer, while the push-pull method involves the injection (push) and extraction (pull) of a radon-free test solution from a single well. In the presence of DNAPL, radon concentrations during the pull phase are retarded, with retardation manifested in greater dispersion of radon concentrations relative to a conservative tracer. The utility of these methods was investigated in the laboratory using a physical aquifer model (PAM). Static and push-pull tests were performed before and after contamination of the PAM sediment pack with trichloroethene (TCE), and after alcohol cosolvent flushing and pump-and-treat remediation. Numerical simulations were used to estimate the retardation factor for radon in push-pull tests. Radon partitioning was observed in static and push-pull tests conducted after TCE contamination. Calculated TCE saturations ranged up to 1.4% (static test) and 14.1% (push-pull test). Post-remediation tests showed decreases in TCE saturations. The results show that radon is sensitive to changes in DNAPL saturation in space and time. However, the methods are sensitive to DNAPL saturation heterogeneity, test location, sample size, and test design. The influence of these factors on test results, as well as the apparent overestimation of the retardation factor in push-pull tests, warrant further investigation. Comparison of trichloroethylene reductive dehalogenation by microbial communities stimulated on silicon-based organic compounds as slow-release anaerobic substrates Yu SH, Semprini L WATER RESEARCH 36 (20): 4985-4996 DEC 2002 Abstract: Microcosm studies were conducted to demonstrate the effectiveness of tetrabutoxysilane (TBOS) as a slow-release anaerobic substrate to promote reductive dehalogenation of trichloroethylene (TCE). The abiotic hydrolysis of TBOS and tetrakis(2ethylbutoxy)silane (TKEBS), and the biotic transformations of the hydrolysis products from both were also investigated. Comparison of TCE reductive dehalogenation was performed with microbial communities stimulated from three different sites: Site 300 Lawrence Livermore National Laboratory (LLNL), CA, Point Mugu Naval Weapons Facility, CA, and the Evanite site in Corvallis, OR. Poisoned microcosms showed that 1 mol of TBOS slowly and abiotically hydrolyzes to 4 mol of 1-butanol, while the live microcosms showed the 1-butanol ferments to butyrate and/or acetate, producing H-2. The hydrolysis of TBOS and TKEBS was abiotic and not enhanced by biotic processes under the anaerobic conditions of these tests. Hydrogen consumption was correlated with reductive dehalogenation, indicating it served as an electron donor for reductive dehalogenation. TBOS was found to be a slow-release anaerobic substrate to support long-term dechlorination of TCE to ethylene in Point Mugu microcosms, and in the LLNL microcosm bioaugmented with the Evanite culture. Electron mass balances showed most of the electron flow went into the creation of organic acids, especially acetate, and the production of methane. Electron efficiencies for reductive dechlorination were as high as 14% based on the electrons used for, dechlorination to the total electrons associated with the mass of TBOS and TKEBS hydrolyzed. Rates of TBOS hydrolysis increased with greater TBOS concentrations as a light nonaqueous-phase liquids (LNAPL). These results indicate that TBOS has promise as an effective anaerobic substrate for remediating a wide range of CAH concentrations at different CAH contaminated sites. (C) 2002 Elsevier Science Ltd. All rights reserved. Kinetic and inhibition studies for the aerobic cometabolism of 1,1,1-trichloroethane, 1,1-dichloroethylene, and 1,1-dichloroethane by a butane-grown mixed culture Kim Y, Arp DJ, Semprini L BIOTECHNOLOGY AND BIOENGINEERING 80 (5): 498-508 DEC 5 2002 Abstract: Batch kinetic and inhibition studies were performed for the aerobic cometabolism of 1,1,1-trichloroethane (1,1,1-TCA), 1,1-dichloroethylene (1,1-DCE), and 1,1dichloroethane (1,1-DCA) by a butane-grown mixed culture. These chlorinated aliphatic hydrocarbons (CAHs) are often found together as cocontaminants in groundwater. The maximum degradation rates (k(max)) and half-saturation coefficients (K-s) were determined in single compound kinetic tests. The highest k(max). was obtained for butane (2.6 mumol/mg TSS/ h) followed by 1,1-DCE (1.3 mumol/mg TSS/h), 1,1-DCA (0.49 mumol/mg TSS/h), and 1,1,1-TCA (0.19 mumol/mg TSS/ h), while the order of K-s from the highest to lowest was 1,1-DCA (19 muM), butane (19 muM), 1,1,1-TCA (12 muM) and 1,1-DCE (1.5 muM). The inhibition types were determined using direct linear plots, while inhibition coefficients (K-ic. and K-iu) were estimated by nonlinear least squares regression (NLSR) fits to the kinetic model of the identified inhibition type. Two different inhibition types were observed among the compounds. Competitive inhibition among CAHs was indicated from direct linear plots, and the CAHs also competitively inhibited butane utilization. 1,1-DCE was a stronger inhibitor than the other CAHs. Mixed inhibition of 1,1,1-TCA, 1,1-DCA, and 1,1-DCE transformations by butane was observed. Thus, both competitive and mixed inhibitions are important in cometabolism of CAHs by this butane culture. For competitive inhibition between CAHs, the ratio of the K-s values was a reasonable indicator of competitive inhibition observed. Butane was a strong inhibitor of CAH transformation, having a much lower inhibition coefficient than the Ks value of butane, while the CAHs were weak inhibitors of butane utilization. Model simulations of reactor systems where both the growth substrate and the CAHs are present indicate that reactor performance is significantly affected by inhibition type and inhibition coefficients. Thus, determining inhibition type and measuring inhibition coefficients is important in designing CAH treatment systems. (C) 2002 Wiley Periodicals, Inc. Push-pull partitioning tracer tests using radon-222 to quantify non-aqueous phase liquid contamination Davis BM, Istok JD, Semprini L JOURNAL OF CONTAMINANT HYDROLOGY 58 (1-2): 129-146 SEP 2002 Abstract: Naturally occurring radon in groundwater can be used as an in situ partitioning tracer for locating and quantifying non-aqueous phase liquid (NAPL) contamination in the subsurface. When combined with the single-well, push-pull test, this methodology has the potential to provide a low-cost alternative to inter-well partitioning tracer tests. During a push-pull test, a known volume of test solution (radon-free water containing a conservative tracer) is first injected ("pushed") into a well; flow is then reversed and the test solution/groundwater mixture is extracted ("pulled") from the same well. In the presence of NAPL radon transport is retarded relative to the conservative tracer. Assuming linear equilibrium partitioning, retardation factors for radon can be used to estimate NAPL saturations. The utility of this methodology was evaluated in laboratory and field settings. Laboratory push-pull tests were conducted in both non-contaminated and trichloroethene NAPL (TCE)-contaminated sediment. The methodology was then applied in wells located in non-contaminated and light non-aqueous phase liquid (LNAPL)-contaminated portions of an aquifer at a former petroleum refinery. The method of temporal moments and an approximate analytical solution to the governing transport equations were used to interpret breakthrough curves and estimate radon retardation factors; estimated retardation factors were then used to calculate TCE saturations. Numerical simulations were used to further investigate the behavior of the breakthrough curved. The laboratory and field push-pull tests demonstrated that radon retardation does occur in the presence of TCE and LNAPL and that radon retardation can be used to calculate TCE saturations. Laboratory injection-phase test results in TCEcontaminated sediment yielded radon retardation factors ranging from 1.1 to 1.5, resulting in calculated TCE saturations ranging from 0.2 to 0.9%. Laboratory extractionphase test results in the same sediment yielded a radon retardation factor of 5.0, with a calculated TCE saturation of 6.5%. Numerical simulation breakthrough curves provided reasonably good matches to the approximate analytical solution breakthrough curves. However, non-equilibrium radon partitioning and heterogeneous TCE distributions may affect the retardation factors and TCE saturation estimates. (C) 2002 Elsevier Science B.V. All rights reserved. A combined method for determining inhibition type, kinetic parameters, and inhibition coefficients for aerobic cometabolism of 1,1,1-trichloroethane by a butane-grown mixed culture Kim Y, Arp DJ, Semprini L BIOTECHNOLOGY AND BIOENGINEERING 77 (5): 564-576 MAR 5 2002 Abstract: A combined method for determining inhibition type, kinetic parameters, and inhibition coefficients is developed and presented. The method was validated by applying it to data obtained from batch kinetics of the aerobic cometabolism of 1,1,1-trichloroethane (1,1,1TCA) by a butane-grown mixed culture. The maximum degradation rates (k(max)) and half-saturation coefficients (K-s) were independently determined in single compound tests, and compared with those obtained from inhibition tests. The inhibition type was determined using direct linear plots at various substrate and inhibitor concentrations. Kinetic parameters (k(max) and K-s) and inhibition coefficients (K-ic and K-iu) were determined by nonlinear least squares regression (NLSR) fits of the inhibition model determined from the direct linear plots. Initial guesses of the kinetic parameters for NLSR were determined from linearized inhibition equations that were derived from the correlations between apparent maximum degradation rates (k(max)(app)) and/or the apparent half-saturation coefficient (K-s(app)) and the k(max), K-s, and inhibitor concentration (I-L) for each inhibition equation. Two different inhibition types were indicated from the direct linear plots: competitive inhibition of 1,1,1-TCA on butane degradation, and mixed inhibition of 1,1,1-TCA transformation by butane. Good agreement was achieved between independently measured k(max), and K-s values and those obtained from both NLSR and the linearized inhibition equations. The initial guesses of all the kinetic parameters determined from linear plots were in the range of the values estimated from NLSR analysis. Overall the results show that use of the direct linear plot method to identify the inhibition type, coupled with initial guesses from linearized plots for NLSR analysis, results in an accurate method for determining inhibition types and coefficients. Detailed studies with pure cultures and purified enzymes are needed to further demonstrate the utility of this method. (C) 2002 John Wiley Sons, Inc. Bioaugmentation of butane-utilizing microorganisms to promote cometabolism of 1,1,1-trichloroethane in groundwater microcosms Jitnuyanont P, Sayavedra-Soto LA, Semprini L BIODEGRADATION 12 (1): 11-22 2001 Abstract: The transformation of 1,1,1-trichloroethane (1,1,1-TCA) in ioaugmented and nonaugmented microcosms was evaluated. The microcosms contained roundwater and aquifer materials from a test site at Moffett Field, Sunnyvale, CA. The initial inoculum for bioaugmentation was a butane-utilizing enrichment from the subsurface of the Hanford DOE site. The non-augmented microcosm required 80 days of incubation before butane-utilization was observed while the augmented microcosms required 3 days. Initially the augmented microcosms were effective in transforming 1,1,1-TCA, but their transformation ability decreased after prolonged incubation. The non-augmented microcosms initially showed limited 1,1,1-TCA transformation but improved with time. After 440 days, both the non-augmented and augmented microcosms had similar transformation yields (0.04 mg 1,1,1-TCA/mg butane) and had similar microbial composition (DNA fingerprints). Subsequent microcosms, when bioaugmented with a Hanford enrichment that was repeatedly grown in 100% mineral media, did not effectively grow or transform 1,1,1-TCA under groundwater nutrient conditions. Microcosm tests to study the effect of mineral media on transformation ability were performed with the Hanford enrichment. Microcosms with 50% mineral media in groundwater most effectively utilized butane and transformed 1,1,1-TCA, while microcosms with groundwater only and microcosms with 5% mineral media in groundwater lost their 1,1,1-TCA transformation ability. DNA fingerprinting indicated shifts in the microbial composition with the different mineral media combinations. Successful bioaugmentation was achieved by enriching butane-utilizers from Moffett Field microcosms that were effective in groundwater with no mineral media added. The results suggest that successful in-situ bioaugmentation might be achieved through the addition of enriched cultures that perform well under subsurface nutrient conditions. In situ anaerobic transformation of trichlorofluoroethene in trichloroethenecontaminated groundwater Hageman KJ, Istok JD, Field JA, Buscheck TE, Semprini L ENVIRONMENTAL SCIENCE & TECHNOLOGY 35 (9): 1729-1735 MAY 1 2001 Abstract: Methods are needed to obtain in situ information on the transformation rates of trichloroethene (TCE), the most commonly detected organic groundwater contaminant. The objective of this research was to investigate the potential for determining TCE transformation rates in groundwater by measuring the transformation rate of its fluorinated surrogate, trichlorofluoroethene (TCFE). To explore this hypothesis, the in situ transport behavior, transformation pathway, and transformation rate of injected TCFE were determined in TCE-contaminated groundwater using single-well, push-pull tests. Although transport behavior varied between wells, TCFE, dichlorofluoroethene (DCFE), and TCE were transported similarly to each other. In the shallow water-hearing zone, TCFE was reductively dechlorinated to cis-DCFE, trans-DCFE, and (E)-1-chloro2-fluoroethene (CFE), while co-injected TCE was concurrently transformed to cisdichloroethene (DCE), trans-DCE, 1,1-DCE, and a trace amount of chloroethene (CE). With added formate and the injected TCFE concentration being a factor of 20 higher than that of TCE, the TCFE transformation rate ranged from 0.053 to 0.30 mu mol/L-day, while that of TCE ranged from 0.009 to 0.012 mu mol/L-day. Without added formate, the TCFE transformation rate decreased to 0.036 mu mol/L-day. In the deeper water-bearing zone, TCFE transformation occurred only after a lag time of 55 days with added formate. No TCFE transformation occurred in groundwater that had not previously been exposed to TCE. The potential applicability for TCFE as an in situ transport and transformation surrogate for TCE was demonstrated. Chlorinated solvent cometabolism by butane-grown mixed culture Kim Y, Arp DJ, Semprini L JOURNAL OF ENVIRONMENTAL ENGINEERING-ASCE 126 (10): 934-942 OCT 2000 Abstract: A survey of aerobic cometabolism of chlorinated aliphatic hydrocarbons by a butane- grown mixed culture was performed. The transformation of 1,1-dichloroethylene (1,1DCE) and cis-1,2-dichloroethylene (c-DCE) required O-2 and was inhibited by butane and inactivated by acetylene, indicating that a monooxygenase enzyme was Likely involved in the transformations. The initial transformation rates and the quantities of chlorinated aliphatic hydrocarbons transformed were inversely proportional to the chlorine contents within each group of chlorinated methanes, ethanes, and ethenes. Lower quantities of chloroform were transformed than chloromethane and dichloromethane, but chloroform transformation resulted in much higher cell inactivation. For the ethane group, chloroethane was most effectively transformed but caused significant cell inactivation. Dior trichloroethanes that have all chlorines on one carbon were more effectively transformed and caused less cell inactivation than the isomers that have chlorine on both carbons. For chlorinated ethenes, 1,1-DCE was most rapidly transformed, whereas trans- 1,2-dichloroethylene was not transformed. Vinyl chloride was transformed to the greatest extent, and very Limited transformation of trichloroethylene was observed. The I,1-DCE transformation caused greater cell inactivation than the transformation of the other chlorinated ethenes. Chloride release studies showed nearly complete oxidative dechlorination of chlorinated methanes and chloroethane, vinyl chloride and c-DCE (86% similar to 100%). and incomplete dechlorination of 1,1-dichloroethane, 1,2-dichloroethane, 1,1,1-trichloroethane, 1,1,2trichloroethane, and 1,1-DCE (37% similar to 75%) was observed. Laboratory, field and modeling studies of radon-222 as a natural tracer for monitoring NAPL contamination Semprini L, Hopkins OS, Tasker BR TRANSPORT IN POROUS MEDIA 38 (1-2): 223-240 JAN 2000 Abstract: The recently developed natural radon tracer method has potential as a rapid, low-cost, nondestructive, and noninvasive method for quantifying NAPL contamination. In the subsurface, radon-222 (radon) is produced by the decay of naturally occurring radium226 contained in the mineral fraction of aquifer solids. In groundwater radon occurs as a dissolved gas, with a half-life of 3.83 days. In the absence of NAPL, the radon concentration in groundwater quickly reaches a maximum value that is determined by the mineral composition of the aquifer solids, which controls the rate of radon emanation. In the presence of NAPL, however, the radon concentration in the groundwater is substantially reduced due to the preferential partitioning of radon into the organic NAPL phase. A simple equilibrium model and supporting laboratory studies show the reduction in radon concentration can be quantitatively correlated with residual NAPL saturation. Thus, by measuring the spatial distribution in radon it may be possible to identify locations where residual NAPL is present and to quantify the NAPL saturation. When the basic processes of partitioning, radon emanation from the aquifer solids, and first-order decay are incorporated into an advective/dispersive transport model, good agreement is obtained with the results of laboratory and field experiments. Model sensitivity analyses shows many factors can contribute to the radon concentration response, including the length of the NAPL zone, NAPL saturation, groundwater velocity, porosity, and radon emanation. Thus, care must be taken when applying the radon method to locate and quantify NAPL contamination in the subsurface. Diversity in butane monooxygenases among butane-grown bacteria Hamamura N, Storfa RT, Semprini L, Arp DJ APPLIED AND ENVIRONMENTAL MICROBIOLOGY 65 (10): 4586-4593 OCT 1999 Abstract: Butane monooxygenases of butane-grown Pseudomonas butanovora, Mycobacterium vaccae JOB5, and an environmental isolate, CF8, were compared at the physiological level. The presence of butane monooxygenases in these bacteria was indicated by the following results. (i) O-2 was required for butane degradation. (ii) 1-Butanol was produced during butane degradation. (iii) Acetylene inhibited both butane oxidation and 1-butanol production. The responses to the known monooxygenase inactivator, ethylene, and inhibitor, allyl thiourea (ATU), discriminated butane degradation among the three bacteria. Ethylene irreversibly inactivated butane oxidation by P. butanovora but not by RI. vaccae or CF8. In contrast, butane oxidation by only CF8 was strongly inhibited by ATU. In all three strains of butane-grown bacteria, specific polypeptides were labeled in the presence of [C-14]acetylene. The [C-14]acetylene labeling patterns were different among the three bacteria. Exposure of lactate-grown CFS and P. butanovora a and glucose-grown M. vaccae to butane induced butane oxidation activity as well as the specific acetylene-binding polypeptides. Ammonia was oxidized by all three bacteria. P. butanovora oxidized ammonia to hydroxylamine, while CF8 and M. vaccae produced nitrite. All three bacteria oxidized ethylene to ethylene oxide. Methane oxidation was not detected by any of the bacteria. The results indicate the presence of three distinct butane monooxygenases in butane-grown. P. butanovora, M. vaccae, and CF8. Anaerobic biotransformation of trichlorofluoroethene in groundwater microcosms Vancheeswaran S, Hyman MR, Semprini L ENVIRONMENTAL SCIENCE & TECHNOLOGY 33 (12): 2040-2045 JUN 15 1999 Abstract: The biological reduction of trichlorofluoroethene (TCFE) was investigated in anaerobic groundwater microcosms. TCFE was reductively dehalogenated by microorganisms to produce three dichlorofluoroethene isomers, with cis-1,2-dichlorofluoroethene (c-DCFE) being the main isomer formed. Further sequential biological transformation of these compounds to mono-chlorofluoroethene isomers was incomplete and occurred at much slower rates. The rates of TCFE reduction were compared to the rates of reduction of two common chlorinated solvents, perchloroethene (PCE) and trichloroethene (TCE), when present at similar concentrations. Aqueous concentrations ranged from 7.0 to 14.0 mg/L for TCFE and from 7.5 to 15.0 mg/L for PCE and TCE. Similar rates of PCE and TCE transformation relative to TCFE were observed in single-compound tests (PCE, TCE, and TCFE in separate microcosms) and when the contaminants were present together as mixtures in the microcosms. The close similarities between the time course and kinetics of TCFE degradation and the degradation of both PCE and TCE, when present at comparable initial concentrations, suggest that TCFE could potentially be used as a benign reactive tracer to measure in-situ rates of PCE and TCE transformation in contaminated environments. Abiotic and biological transformation of tetraalkoxysilanes and trichloroethene/cis1,2-dichloroethene cometabolism driven by tetrabutoxysilane-degrading microorganisms Vancheeswaran S, Halden RU, Williamson KJ, Ingle JD, Semprini L ENVIRONMENTAL SCIENCE & TECHNOLOGY 33 (7): 1077-1085 APR 1 1999 Abstract: Attenuation of silicon-based organic compounds (tetraalkoxysilanes) by abiotic hydrolysis and biological mineralization was investigated. At Lawrence Livermore National Laboratory site 300, tetraalkoxysilanes are present along with trichloroethene (TCE) as subsurface contaminants. Under abiotic conditions, the alkoxysilanes such as tetrabutoxysilane (TBOS) and tetrakis(2-ethylbutoxy)silane (TKEBS) hydrolyze to lbutanol and 2-ethylbutanol, respectively, and silicic acid. The rates of hydrolysis of TBOS and TKEBS were determined to evaluate the significance of the hydrolysis reaction in the attenuation process, and typical rates at pH 7, 30 degrees C, and 28 mu mol/L initial concentration were 0.32 and 0.048 mu mol/L/day, respectively. The TBOS hydrolysis reaction was observed to be acid- and base-catalyzed and independent of temperature from 15 to 30 degrees C. All hydrolysis experiments were conducted at concentrations above the solubility limit of TBOS and TKEBS, and the rate of hydrolysis increased with concentration of TBOS or TKEBS. An aerobic microbial culture from the local wastewater treatment plant that could grow and mineralize the alkoxysilanes was enriched. The enriched culture rapidly hydrolyzed TBOS and TKEBS and grew on the hydrolysis products. The microorganisms grown on TBOS cometabolized TCE and cis1,2-dichloroethene (c-DCE). TCE and c-DCE degradation was inhibited by acetylene, indicating that a monooxygenase was involved in the cometabolism process. Acetylene did not inhibit the hydrolysis of TBOS or the utilization of l-butanol, indicating that the above monooxygenase enzyme was not involved in the degradation of TBOS. Chloroform cometabolism by butane-grown CF8, Pseudomonas butanovora, and Mycobacterium vaccae JOB5 and methane-grown Methylosinus trichosporium OB3b Hamamura N, Page C, Long T, Semprini L, Arp DJ APPLIED AND ENVIRONMENTAL MICROBIOLOGY 63 (9): 3607-3613 SEP 1997 Abstract: Chloroform (CF) degradation by a butane-grown enrichment culture, CF8, was compared to that by butane-grown Pseudomonas butanovora and Mycobacterium vaccae JOB5 and to that by a known CF degrader, Methylosinus trichosporium OB3b. All three butanegrown bacteria were able to degrade CF at rates comparable to that of M. trichosporium. CF degradation by all four bacteria required O-2. Butane inhibited CF degradation by the butane-grown bacteria, suggesting that butane monooxygenase is responsible for CF degradation. P. butanovora required exogenous reductant to degrade CF, while CF8 and M. vaccae utilized endogenous reductants. Prolonged incubation with CF resulted in decreased CF degradation. CF8 and P. butanovora were more sensitive to CF than either M. trichosporium or M. vaccae. CF degradation by all three butane-grown bacteria was inactivated by acetylene, which is a mechanism-based inhibitor for several monooxygenases. Butane protected all three butane-grown bacteria from inactivation by acetylene, which indicates that the same monooxygenase is responsible for both CF and butane oxidation. CF8 and P. butanovora were able to degrade other chlorinated hydrocarbons, including trichloroethylene, 1,2-cis-dichloroethylene, and vinyl chloride. In addition, CF8 degraded 1,1,2-trichloroethane. The results indicate the potential of butane-grown bacteria for chlorinated hydrocarbon transformation. Model simulations in support of field scale design and operation of bioremediation based on cometabolic degradation Lang MM, Roberts PV, Semprini L GROUND WATER 35 (4): 565-573 JUL-AUG 1997 Abstract: This paper addresses questions fundamental to the design and operation of aquifer bioremediation based on cometabolic degradation. A model of a full-scale, in situ system for bioremediation of chlorinated ethenes relying on cometabolic degradation was developed and applied to a hypothetical aquifer being considered for a large-scale field demonstration of in situ bioremediation with recirculation. The model was used to identify feasible substrate (electron donor and electron acceptor) delivery schedules. Trichloroethylene (TCE) was the target contaminant. Methane and phenol were considered as electron donors. The delivery of the electron donors and the electron acceptor, oxygen, was varied to evaluate the rate and extent of bioremediation under different substrate delivery schedules. Maximum removal of TCE was predicted when substrates are delivered at ratios near the stoichiometric requirement of electron donor and acceptor for net microbial growth. Additionally, the decrease in TCE removal that results from using substrate delivery schedules other than those achieving the maximum removal of TCE was quantified. This decrease was greater for the methane-oxygen system because the two gaseous substrates compete for transfer into the recirculated ground water. If one substrate is introduced in excess of the amount required for net microbial growth, it accumulates, thus limiting the ability to introduce the second substrate. This imbalance both limits the introduction of the second substrate and accelerates the accumulation of the substrate added in excess. The phenol-oxygen system is less sensitive to deviation away from the best observed substrate delivery schedule because phenol is a relatively soluble liquid and its introduction does not compete with the mass transfer of oxygen. Strategies for the aerobic co-metabolism of chlorinated solvents Semprini L CURRENT OPINION IN BIOTECHNOLOGY 8 (3): 296-308 JUN 1997 Abstract: Recent field and laboratory studies have evaluated the potential for aerobic cometabolism of chlorinated solvents. Different co-metabolic substrates and different methods of application have been tried, including growing indigenous microbes in situ, and injecting into the soil subsurface strains grown in subsurface reactors for their ccmetabolic potential. There is much potential for using co-metabolism for treating a broad range of chlorinated aliphatic hydrocarbons. Recirculation wells have potential for adding soluble cc-metabolic substrates (i.e. phenol and toluene) into contaminated aquifers, while direct addition of gaseous substrates (i.e. methane and propane) into aquifers also holds promise. Aromatic substrates (phenol and toluene) are best used for treatment of chlorinated ethenes, whereas gaseous co-metabolic substrate (methane and propane) are better suited for the treatment of chlorinated methanes and ethanes. Many factors can enhance co-metabolic transformations, such as nutrients and available energy sources. (C) Current Biology Ltd. Acetylene Inhibition of Trichloroethene and Vinyl Chloride Reductive Dechlorination George Pon, Michael R. Hyman, and Lewis Semprini* ENVIRONMENTAL SCIENCE AND TECHNOLOGY Volume 37 pp3181-3188 (2003) Abstract: Kinetic studies reported here have shown that acetylene is a potent reversible inhibitor of reductive dehalogenation of trichloroethene (TCE) and vinyl chloride (VC) by a mixed dehalogenating anaerobic culture. The mixed culture was enriched from a contaminated site in Corvallis, OR, and exhibited methanogenic, acetogenic, and reductive dehalogenation activities. The H2-fed culture transformed TCE to ethene via cisdichloroethene (c-DCE) and VC as intermediates. Batch kinetic studies showed acetylene reversibly inhibited reduction of both TCE and VC, and the levels of inhibition were strongly dependent on acetylene concentrations in both cases. Acetylene concentrations of 192 and 12 M, respectively, were required to achieve 90% inhibition in rates of TCE and VC transformation at an aqueous concentration of 400 M. Acetylene also inhibited methane production (90% inhibition at 48 M) but did not inhibit H2-dependent acetate production. Mass balances conducted during the studies of VC inhibition showed that acetogenesis, VC transformation to ethene, and methane production were responsible for 52%, 47%, and 1% of the H2 consumption, respectively. The results indicate that halorespiration is the dominant process responsible for VC and TCE transformation and that dehalorespiring organisms are the target of acetylene inhibition. Acetylene has potential use as a reversible inhibitor to probe the biological activities of reductive dechlorination and methanogenesis. It can be added to inhibit reactions and then removed to permit reactions to proceed. Thus, it can be a powerful tool for investigating intrinsic and enhanced anaerobic remediation of chloroethenes at contaminated sites. The results also suggest that acetylene produced abiotically by reactions of chlorinated ethenes with zero-valent iron could inhibit the biological transformation of VC to ethene.