In Situ and Ex Situ Bioremediation Technologies for Groundwater Contaminated with Chlorinated Solvents



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In Situ and Ex Situ Bioremediation Technologies for Groundwater Contaminated with Chlorinated Solvents Michelle M. Lorah, U.S. Geological Survey Emily Majcher, Duane Graves, Geosyntec Consultants John Wrobel, Aberdeen Proving Ground

In situ test area, Seep 3-4W West Branch Plumes Canal Creek Groundwater Treatment Plant- Ex situ test East Branch Plume Canal Creek Area, Aberdeen Proving Ground, Maryland East Branch Plume

Chlorinated VOCs at West Branch Canal Creek and their anaerobic degradation pathways PtCA 1122- TeCA HCA PCE Parent VOCs in orange TCE CT Chlorinated ethanes: HCA= hexachloroethane PtCA= pentachloroethane 1122TeCA= 1,1,2,2-tetrachloroethane 112TCA 12DCE CF Chlorinated ethenes: PCE= tetrachloroethene TCE= trichloroethene 12DCA VC CO 2 MeCl Chlorinated methanes: CT= carbon tetrachloride CF= chloroform CA Ethane Ethene Methane

Natural Attenuation in Wetland

W W B23+ B30 On emon th TT eca eca +TeCA inc 1) inc1) 2) ubatio reduction pre-expo n: sure +TeC WB23 WB2+ +TeCA TeC TeC A WB2+ TeC A+TCA +DCE +DCE +VC +VC Te Te T+ CA + TCA D D + CA CA CE + V VC VC et et hene hene D ad a CE2 cetate ndh+ CE2 cetate ndh+ TCA2acetateandH+ DCEacetatandH+ VC+aceta 2 e VC+aceta V+ te 15 15 12 5 2 H H 2 Wes tbran chcon sortium Development of WBC-2 Microbial Consortium 8 liter 20 liter 100 liter

WBC-2 Substrate Range Chlorinated Ethanes, Ethenes, Methanes y 0.6 0.08 5 Concentration (mmoles) 0.5 0.4 0.3 0.2 0.1 ethene 1,1,2,2-TECA 1,1,2-TCA cis-1,2-dce trans-1,2-dce VC Ethene TCE Chlorinated Methanes (mmoles) 0.06 0.04 0.02 methane CTC CF DCM CM Methane 4 3 2 1 Methane (mmoles) 0 0 2 4 6 8 10 12 14 Days 0 0 0 5 10 15 20 25 30 Sediment-free culture with consistent dechlorination ability Regularly fed 5 mg/l TeCA, 112TCA, cis12dce, and lactate and ethanol (e - donors) Degrades carbon tetrachloride and chloroethene/ethane degradation unaffected WBC-2 degradation stable since 2003 Days

WBC-2 Microbial Composition Dehalococcoides Desulfovibrio WBC-2 Acidaminococcaceae (putative) No match Contains at least 17 and 21 distinct Bacteria phylotypes based on DGGE and clone library analysis, respectively Syntrophus Trichlorobacter Dehalobacter Spirochete Dehalococcoides sp. CBDB1 No match Dehalococcoides ethenogenes 195 Dehalococcoides sp. CBDB1 No match Acetobacterium (40%), Dehalobacter (19%), and Acidaminococcacea (9%) most prominent; Dehalococcodies 5% Diversity thought to support substrate range

Pilot-Test Area 3-4W3 Chloroform Tetrachloroethene

Reactive Mat Design and Monitoring Re-amendment System Below surface microwells Single-hole multilevel diffusion samplers and multilevel wells Pea Gravel Sand, Peat, Compost, Chitin, WBC2 Passive diffusion bags Sand, Peat, Compost, ZVI (option for CT) Geotextile Hydraulic Gradient (Artesian) Seep Area Wetland Sediment (10-15 ft thick) Aquifer (~40 ft thick) NOT TO SCALE

Flow-Through Column Tests - Biomat Simulate biomat layers Continuous upflow columns Establish similar discharge rates/concentrations to field Evaluate sediment only, compost/peat mix, and iron layer before compost/peat mix Measure VOCs, redox, culture behavior, hydraulic properties

Degradation of VOCs by WBC-2 seeded sediment in columns WBC-2 only WBC-2 and lactate

Bioreactive Mat Construction Sprayed WBC2 between each wheelbarrow load of material

Reactive Mat Location and Monitoring Network Top layer diffusion sampling grid

Reactive Mat VOC Removal Consistent vertical profiles of VOC reduction through mat, increased ethene/ethane 95 to 99% mass removal Depth (FT BLS) -1 1 3 5 7 9 11 13 um Concentration 0 2 4 6 8 VC cdce TCE PCE TeCA PCA HCA ETHENE ETHANE Depth (FT BLS) -1 1 3 5 7 9 11 13 um Concentration 0 50 100 150 MeCl CF CT

Sustainability of Degradation (A) (B) September 2008 VOCs non-detect (A) Consistent upward flow of VOC to mat over 1 year monitoring period (B) Consistent VOC degradation within reactive mat over 1 year monitoring period October 2008 sampling- still effective 4 years after emplacement.

Application of WBC-2 in an Anaerobic Bioreactor for the Canal Creek Groundwater Treatment Plant

Advantages of Fixed Film Anaerobic Bioreactor System Mechanically simple with low energy costs and maintenance Very low production of waste solids (sludge) Highly resistant to upsets and toxic shock Insignificant pretreatment of water (no bag filters needed) An anaerobic culture is available that can degrade the suite of chlorinated VOCs in the treatment plant influent WBC-2 has been grown previously on a common bioreactor support medium in a static batch, where it retained its full dechlorinating ability Extensive testing of WBC-2 in lab and field has shown its robustness and ability to obtain high rates of degradation

Treatment Plant Influent Contaminant (μg/l) (μm) 1,1,2,2-tetrachloroethane (TeCA) 190 1.13 1,1,2-trichloroethane (112TCA) 4.7 0.04 1,,2-dichloroethane (12DCA) 4.2 0.04 Carbon tetrachloride (CT) 15 0.10 Chloroform (CF) 5.6 0.05 Tetrachloroethene (PCE) 2.8 0.02 Trichloroethene (TCE) 130 0.99 Vinyl chloride (VC) 71 1.14 cis- and trans-1,2-dichloroethene (12DCE) 167 1.72 Total VOCs 590 4.5 Maximum Total VOCs 1,000 7.6

Rates for initial VOC= 40 μ M Contaminant Mean rate Constant (day -1 ) Degradation rates in mat columns- Half- Life (hour) Rate (μm/day) 1,1,2,2-tetrachloroethane (TeCA) 3.43 4.85 140 More than 4 times the degradation rate in static culture vessel Chloroform (CF) 2.31 7.20 92 Carbon tetrachloride (CT) 2.83 5.88 113 Tetrachloroethene (PCE) 1.40 11.9 56 Trichloroethene (TCE) 3.04 5.47 122

WBC-2 20L Vessel Filled with EXCEED Polyurethane Foam 2.5 45 VOCs 40 VOCs mmoles 2.0 1.5 1.0 VOCs Spiked: 5 mg/lteca, 112-TCA, cisdce Methane Ethene VOCs VOCs 35 30 25 20 15 Methane mmoles 0.5 10 5 0.0 0 5 10 15 20 25 30 35 40 45 50 55 60 Days 0 TCE cis-12dce trans-12dce VC Ethene TeCA 1,1,2-TCA 1,2-DCA Ethane Methane WBC-2 has been grown previously on a common bioreactor support medium in a static batch, where it retained its full dechlorinating ability

Bioreactor Study Objectives Conduct pre-design testing necessary to evaluate the effectiveness of WBC-2 for treating chlorinated VOCs in a bioreactor application optimal hydraulic retention time to achieve efficient VOC degradation and meet discharge criteria optimal electron donor nutrient and electron donor utilization rates oxygen consumption rate and design constraints to maintain anaerobic conditions in the bioreactor typical equilibration/growth rate for WBC-2 following a change in flow or upset to the system sludge production rates and handling requirements

Bench Test Bioreactor Bicarbonate Stock Solution Argon Nutrient Stock Solution Electron Donor Solution Sampling Ports ORP Electrode Peristaltic Pump Bioreactor Vessel 1 Peristaltic Pump Bioreactor Vessel 2 Bioreactor Vessel 3 Settled Sludge Sludge Separation Clarified Effluent Clarified Effluent internal info: path, date revised, author Groundwater Equalization Tank equalization vessel to mix influent with Schematic nutrients Diagram and of the donor 1-L pretreatment fixed film bioreactor Bioreactor System Two 1-L fixed film bioreactors for VOC removal Figure a 1-L separatory funnel for solids separation 4.1 KNOXVILLE, TENNESSEE 7 April 2009

Bioreactor Bench Test Results

Bioreactor Bench Test Results

Bioreactor: Donor Testing Microcosms show dissolvable packing peanuts (corn starch) and corn syrup are inexpensive possibilities Currently testing a lower grade lactate Small-scale hydrogen generators (electrolysis cells) have been made and will be placed in-line in one of the bioreactor bench tests

Conclusion: in situ bioremediation application of WBC-2 was successful and preliminary results show promise for ex situ application in bioreactors USGS Other Current Studies: WBC-2 application for RDX and perchlorate bioremediation, White Sands Missile Range, New Mexico WBC-2 application for chlorobenzenes bioremediation in wetland area, Standard Chlorine Superfund site, Delaware Cooperative Research and Development Agreement (CRADA) between USGS and GeoSyntec.

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