Sustained In Situ DetoxificaTon of Priority Chloroorganic Pollutants - PowerPoint PPT Presentation

Sustained In Situ DetoxificaTon of Priority Chloroorganic Pollutants Frank Löffler Center for Environmental Microbiology Department of Microbiology Department of Civil & Environmental Engineering Bioscience Division, Oak Ridge NaKonal Laboratory

Saltwater Oceans 97.5% Freshwater 2.5% Ice Caps and Glaciers 79% Groundwater 20% 1% Water in Lakes 52% Water in Soil 38% Water in Rivers Water Vapor Water in Organisms in Atmosphere

Contaminated Sites in the U.S. 1,322 Superfund sites 3,747 RCRA sites >450,000 Brownfields 53% 55% of a Superfund or 166 M ILLION 3 OF ALL Live Including a RCRA CorrecKve U.S. OF THE C HILDREN Within P EOPLE M ILES P OPULATION AcKon Site, Equal to UNDER A GE 5 ... and in Europe EPA, 2013 Majority of sites impacted with chlorinated compounds 3

Chlorinated Ethenes as Respiratory Electron Acceptors 2[H] 2[H] 2[H] 2[H] -173 -169 -139 -149 H Cl HCl HCl HCl HCl Energy Energy Energy Energy Organohalide RespiraTon (Respiratory Reductive Dechlorination) ∆ G°' kJ/reaction with H 2 as the electron donor 4

Chlorinated Ethenes are Major Risk Drivers H Cl PCE TCE cis -DCE VC Ethene 5 ppb 5 ppb 7 ppb 2 ppb Human carcinogen Human carcinogen Parkinson’s disease Premature birth 5

Contaminant Source Zones and Plumes • Suspected or proven human carcinogens • Increased risk of Parkinson Disease • Premature birth Unsaturated TCE Zone Capillary Fringe Water Table Low Permeability Groundwater Lenses Flow Dissolved-Phase Plume DNAPL Pools Confining Layer 6

PopulaTons Involved in ReducTve DechlorinaTon of Chlorinated Ethenes Dehalococcoides mccartyi Geobacter lovleyi, Dehalobacter, Sulfurospirillum, Desulfuromonas, Desulfitobacterium He et al. 2003, Nature, 424:62 Löffler et al. 2013, IJSEM, 63:625 7

Novel Dechlorinators from PrisTne Sources 160 VC and ethene µmol/boWle VC 140 Respiked VC Microcosms 120 100 Ethene 80 60 40 20 Grape pomace 0 0 20 40 60 80 100 120 140 160 180 200 220 compost Enrichment Time (days) Cultures 8

PrisTne Environments are Reservoirs for Novel RDase Genes 16S Amplicon Sequencing Metagenome Sequencing 1 st VC 2 nd VC Enrichment AddiTon AddiTon Culture Others 32.1 16.5 VadinCA02 1.3 2.8 Clostridium 10.1 13.6 Pelotomaculum 1 1.6 Bacteroidetes 4.5 6.8 Unclassified WPS-2 1.8 9.4 Dehalogenimonas 47.4 46.1 Oscillospira 0.1 0.08 Geobacter 0.1 0.22 Treponema 0.7 2.6 Dehalococcoides 0 0 Acetobacterium 0.9 0.3 9 Molenda et al. 2016, AEM 82:40

Phylogeny of Novel VC Dechlorinator Organohalide-Respiring Chloroflexi 10

PCBs CEs 11

1173 GW samples, 111 sites 849 samples: Dhc & Dhgm 65%: Dhgm outnumber Dhc PCBs CEs c DCE 143 159 914 859 53 44 VC Ethene TCE 474 694 1169 902 205 280 1,1- 116 816 DCE Yi et al. ISME J. In Revision 12

Plume Teatment - BioreacTve Barriers BiosKmulaKon BioaugmentaKon MERL Oregon State University 13

Simplified Model of Organohalide RespiraTon H 2 Dehalococcoides 2 H + mccartyi strain BAV1 H + 2 e - R-Cl + H + Corrinoid' ATP Cofactor' R-H + Cl - H + 14

Dhc : Strict Requirement for Corrinoid Dhc strain BAV1 cDCE No B 12 VC cDCE stall ETH Limited B 12 [1 µ g/L] VC stall Complete Sufficient B 12 Dechlorination [25 µ g/L] (Detoxification) 15

Lower Bases Upper ligand Benzimidazole (Bza) type OH OCH 3 OCH 3 N N N N N N CH 3 Corrin ring CH 3 CH 3 N N N N N N CH 3 Nucleobase type N N N CH 3 N SCH 3 N N N N N N N N N N N N NH 2 OH NH 2 NH 2 O O Lower base N SCH 3 N SCH 3 N NH 2 N N N O N N NH N N N NH 2 NH 2 O Phenol type Naphthimidazole N O O CH 3 Dimethylbenzimidazole (DMB) N R = CN = Cyanocobalamin (Vitamin B 12 )

Who Supplies the Corrinoid? Dehalococcoides mccartyi Geobacter lovleyi ? Corrinoid + Corrinoid – 17

Co-Culture Experiments: Corrinoid Producer / Dhc mccartyi Corrinoid Cobamide Dhc producing auxotroph mccartyi microbe Geobacter spp. Strain BAV1 Strain FL2 ? Dhc dechlorination activity? 18

Summary of Co-Culture Experiments Co-Cultures Dhc Growth Corrinoid Producer Dhc Strains Geobacter lovleyi BAV1, FL2 + Geobacter sulfurreducens BAV1, FL2 - Sporomusa sp. BAV1, FL2, GT - Acetobacterium sp. BAV1, FL2 - Clostridium ace?cum BAV1, FL2 - Methanosarcina barkeri BAV1, FL2, GT - Yan et al. 2012. Appl. Environ. Microbiol. 78:6630-6636 Yan et al. 2013. Phil. Trans. R. Soc. B. 368, 20120320 19

Summary of Co-Culture Experiments Co-Cultures Dhc Dhc Growth Growth Corrinoid Producer Dhc Strains with DMB Geobacter lovleyi BAV1, FL2 + + Geobacter sulfurreducens BAV1, FL2 - + Sporomusa sp. BAV1, FL2, GT - + Acetobacterium sp. BAV1, FL2 - + Clostridium ace?cum BAV1, FL2 - ND Methanosarcina barkeri BAV1, FL2, GT - + Yan et al. 2012. Appl. Environ. Microbiol. 78:6630-6636 Yan et al. 2013. Phil. Trans. R. Soc. B. 368, 20120320 20

Lower Bases Benzimidazole (Bza) type OH OCH 3 OCH 3 N N N N N N CH 3 CH 3 CH 3 N N N N N N CH 3 Nucleobase type N N N CH 3 N SCH 3 N N N N N N N N N N N N NH 2 OH NH 2 NH 2 O O N SCH 3 N SCH 3 N NH 2 N N N O N N NH N N N NH 2 NH 2 O Phenol type Naphthimidazole N O O CH 3 N

Lower Base Affects DechlorinaTon DechlorinaTon rates [mmoles Cl - L -1 d -1 ] Strain BAV1 (BvcA) cDCE, VC and ethene [ µ moles/bottle] [DMB]Cba [Ben]Cba Time (days) Yan et al. 2016. ISME J. 10:1092 22

Lower Base Affects DechlorinaTon DechlorinaTon rates [mmoles Cl - L -1 d -1 ] Strain GT (VcrA) cDCE, VC and ethene [ µ moles/bottle] [DMB]Cba [Ben]Cba Time (days) Yan et al. 2016. ISME J. 10:1092 23

Corrinoid QuanTty and Quality MaWer Methanogens N N Pseudo-vitamin B 12 N N ( Methanococcales ) NH 2 cDCE OH N Factor III ( Methanosarcina ) N No B 12 VC ETH OCH 3 N Factor III m ( Clostridium ) N OCH 3 Limited B 12 N Acetogens ( Clostridium ) N [1 µ g/L] CH 3 CH 3 Cobalamin N ( Acetobacterium ) N CH 3 Sufficient B 12 p -Cresolyl cobamide O CH 3 ( Sporomusa ) [25 µ g/L] Phenolyl cobamide O ( Sporomusa ) SRB CH 3 N ( Desulfobulbus ) N 24

Corrinoid ProducTon Under Different Redox CondiTons Third Creek Site Knoxville, TN Sediment • Metal-manufacturing factory • Third Creek sediment receiving chlorinated solvents (predominantly TCE) from underlying fractures GW Commerce Street Superfund Site MW-7 Williston, Vermont (outside Burlington) • MulK-tenant industrial park • Soil and groundwater contaminants include TCE, cis -DCE, petroleum hydrocarbons and metals (chromium, cadmium, and nickel) 25

Corrinoid ProducTon Under Different Redox CondiTons Corrinoid ExtracTon and IdenTficaTon 26

The Redox Condition Affects Corrinoid Quantity ��������������������������� � �������� � � � � � � � � Lactate Iron Methanogenic Nitrate Sulfate fermenTng reducing reducing reducing 27

The Redox Condition Affects Corrinoid Quality ��������������������������� � �������� � DMB-Cba Bza-Cba � Factor III MeBza-Cba � Phe-Cba P-Phe-Cba � � � � � Lactate Iron Methanogenic Nitrate Sulfate fermenTng reducing reducing reducing 28

Corrinoids Produced by the Community Under Different Redox Conditions Corrinoids(( Lactate( Iron(( Sulfate( Nitrate( Methanogenic( produced( fermen2ng( reducing( reducing( reducing( - + + + + - - + - - - - - + X - - + - - - X - + - - - X - + + - - X + + - - - X 29

Plume Teatment - BioreacTve Barriers CH 4 More BiosKmulaKon Eectron Donor VC stall MERL Oregon State University 30

Team DechlorinaTon Dr. Jun Yan Dr. Fadime Kara Murdoch Dr. Devrim Kaya Yongchao Yin Yongchao Xie Steve Higgins Cindy Swil Microbial Insights, Inc. Burcu Şimşir, Meng Bi, Yi Yang, Dora Ogles-Taggart, BreW Baldwin UTK Chemistry Tuls University Shawn Campagna Natalie Capiro, Kurt Pennell Oak Ridge NaTonal Laboratory University of Toronto Georgia Tech Bob Hehch Elizabeth Edwards Kostas KonstanTnidis