Comparing on-site to off-site biomass collection for Dehalococcoides biomarker gene quantification to predict in situ chlorinated ethene detoxification potential.

Biostimulation and bioaugmentation have emerged as constructive remedies for chlorinated ethene-contaminated aquifers, and a link between Dehalococcoides (Dhc) bacteria and chlorinated ethene detoxification has been established. To quantify Dhc biomarker genes, groundwater samples are shipped to analytical laboratories where biomass is collected on membrane filters by vacuum filtration for DNA extraction and quantitative real-time PCR analysis. This common practice was compared with a straightforward, on-site filtration approach to Sterivex cartridges. In initial laboratory studies with groundwater amended with known amounts of Dhc target cells, Sterivex cartridges yielded one-third of the total DNA and 9-18% of the Dhc biomarker gene copies compared with vacuum filtration. Upon optimization, DNA yields increased to 94 +/- 38% (+/-SD, n = 10), and quantification of Dhc biomarker genes exceeded the values obtained with the vacuum filtration procedure up to 5-fold. Both methods generated reproducible results when volumes containing >10(4) total Dhc target gene copies were collected. Analysis of on-site and off-site biomass collection procedures corroborated the applicability of the Sterivex cartridge for Dhc biomarker quantification in groundwater. Ethene formation coincided with Dhc cell titers of >2 x 10(6) L(-1) and high (i.e., >10(5)) abundance of the vinyl chloride reductive dehalogenase genes vcrA and/or bvcA; however, high Dhc cell titers alone were insufficient to predict ethene formation. Further, ethene formation occurred at sites with high Dhc cell titers but low or no detectable vcrA or bvcA genes, suggesting that other, not yet identified vinyl chloride reductive dehalogenases contribute to ethene formation. On-site biomass collection with Sterivex cartridges avoids problems associated with shipping groundwater and has broad applicability for biomarker monitoring in aqueous samples.

Pilot-scale demonstration of surfactant-enhanced PCE solubilization at the Bachman Road site. 1. Site characterization and test design.

A pilot-scale demonstration of surfactant-enhanced aquifer remediation (SEAR) was conducted to recover dense nonaqueous phase liquid (DNAPL) tetrachloroethene (PCE) from a sandy glacial outwash aquifer underlying a former dry cleaning facility at the Bachman Road site in Oscoda, MI. Part one of this two-part paper describes site characterization efforts and a comprehensive approach to SEAR test design, effectively integrating laboratory and modeling studies. Aquifer coring and drive point sampling suggested the presence of PCE-DNAPL in a zone beneath an occupied building. A narrow PCE plume emanating from the vicinity of this building discharges into Lake Huron. The shallow unconfined aquifer, characterized by relatively homogeneous fine-medium sand deposits, an underlying clay layer, and the absence of significant PCE transformation products, was judged suitable for the demonstration of SEAR. Tween 80 was selected for application based upon its favorable solubilization performance in batch and two-dimensional sand tank treatability studies, biodegradation potential, and regulatory acceptance. Three-dimensional flow and transport models were employed to develop a robust design for surfactant delivery and recovery. Physical and fiscal constraints led to an unusual hydraulic design, in which surfactant was flushed across the regional groundwater gradient, facilitating the delivery of concentrations of Tween 80 exceeding 1% (wt) throughout the treatment zone. The potential influence of small-scale heterogeneity on PCE-DNAPL distribution and SEAR performance was assessed through numerical simulations incorporating geostatistical permeability fields based upon available core data. For the examined conditions simulated PCE recoveries ranged from 94to 99%. The effluent treatment system design consisted of low-profile air strippers coupled with carbon adsorption to trap off-gas PCE and discharge of treated aqueous effluent to a local wastewater treatment plant. The systematic and comprehensive design methodology described herein may serve as a template for application at other DNAPL sites.

Pilot-scale demonstration of surfactant-enhanced PCE solubilization at the Bachman Road site. 2. System operation and evaluation.

A pilot-scale demonstration of surfactant-enhanced aquifer remediation (SEAR) was conducted during the summer of 2000 at the Bachman Road site in Oscoda, MI. Part two of this two-part paper describes results from partitioning and nonpartitioning tracer tests, SEAR operations, and post-treatment monitoring. For this field test, 68 400 L of an aqueous solution of 6% (wt) Tween 80 were injected to recover tetrachloroethene-nonaqueous phase liquid (PCE-DNAPL) from a shallow, unconfined aquifer. Results of a nonreactive tracer test, conducted prior to introducing the surfactant solution, demonstrate target zone sweep and hydraulic control, confirming design-phase model predictions. Partitioning tracer test results suggest PCE-DNAPL saturations of up to 0.74% within the pilot-scale treatment zone, consistent with soil core data collected during site characterization. Analyses of effluent samples taken from the extraction well during SEAR operations indicate that a total of 19 L of PCE and 95% of the injected surfactant were recovered. Post-treatment monitoring indicated that PCE concentrations at many locations within the treated zone were reduced by as much as 2 orders of magnitude from pre-SEAR levels and had not rebounded 450 days after SEAR operations ceased. Pilot-scale costs ($365 900) compare favorably with design-phase cost estimates, with approximately 10% of total costs attributable to the intense sampling density and frequency. Results of this pilot-scale test indicate that careful design and implementation of SEAR can result in effective DNAPL mass removal and a substantial reduction in aqueous concentrations within the treated source zone under favorable geologic conditions

Stimulated microbial reductive dechlorination following surfactant treatment at the Bachman Road site.

A pilot-scale demonstration of surfactant-enhanced aquifer remediation (SEAR) was conducted in July 2000 at the Bachman Road site located in Oscoda, MI. The Bachman aquifer is a shallow, relatively homogeneous, unconfined aquifer formation composed primarily of sandy glacial outwash with relatively low organic carbon content (0.02 wt %). A 6 wt % aqueous solution of Tween 80 (a nonionic, food-grade surfactant) was flushed through a localized dense nonaqueous phase liquid (DNAPL) source zone to recover approximately 19 L of tetrachloroethene (PCE). Post-treatment monitoring revealed PCE concentrations were reduced by up to 2 orders of magnitude within the source zone, and there was no evidence of concentration rebound after more than 450 d. Concentrations of PCE dechlorination products (trichloroethene, cis-1,2-dichloroethene) 450 d after SEAR operations ceased were more than 2 orders of magnitude greater than pretreatment values, suggesting stimulation of native dechlorination activity. Post-treatment monitoring detected increased concentrations of volatile fatty acids generated from the fermentation of residual-level Tween 80 surfactant. These field data suggest that Tween 80 not only induced and maintained anaerobiosis but also provided reducing equivalents to reductively dechlorinating populations present in the oligotrophic Bachman aquifer. Experience from this site supports application of staged treatment strategies that couple SEAR and microbial reductive dechlorination to enhance mass removal and reduce contaminant mass flux emanating from treated source zones.