1. (Total and)Methyl Mercury Export from Denitrifying Bioreactors in Tile-Drained Fields of Central Illinois Robert J.M. Hudson, NRES Richard A.C. Cooke, ABE University of Illinois at Urbana-Champaign

2. Acknowledgements Illinois Sustainable Technology Center Funding for the project Patience with our pace Brian Vermillion (foremerly NRES) MeHg analysis Richard Cooke group Access to field sites and assistance with sampling Expertise

3. A Tale of Two Elements Both have complex biogeochemical cycles involving chemical species in multiple phases and oxidation states: Nitrogen Nitrogen N 2 (g), NO 3 - (aq), NH 4 + (aq), Soil Organic N, etc.Mercury Hg(g), Hg 2+ (aq), CH 3 Hg + (aq ), HgS(s), etc. A Tale of Two Elements

4. Pollutant Sources and Impacts Nitrogen Non-point source pollutant derived from fertilizer overuse Nitrate exported from agricultural fields impacts: Gulf of Mexico ecosystem (hypoxia) Water quality for communities that obtain drinking water from rivers in agricultural areasMercury Non-point source pollutant generated via coal combustion and waste incineration Methylmercury strongly bioaccumulates in food webs: Fish consumption is main source of human exposure to Hg. Leading cause of freshwater fish consumption advisories in the U.S. Significant percentage of U.S. women of childbearing age have blood Hg higher than the USEPA “Threshold Level”. A Tale of Two Elements

5. Anaerobic Processing Key processes that determine the environmental impacts of each element occur in anaerobic environments: Denitrification (alleviates impacts) Denitrification (alleviates impacts) NO 3 - (aq)  NO 2 - (aq)  N 2 (g) Mercury methylation (greatly increases impacts) Hg 2+ (aq)  CH 3 Hg + (aq) A Tale of Two Elements

6. O2O2 Respiration Photosynthesis Reduction O 2 NO 3 - MnO 2 (s) FeO x (s) SO 4 2- Sediments &Hyporheic Groundwater Hg(SH) 2 Methylation MeHg NO 3 - H 2 O N 2 Mn 2+ Fe 2+ H 2 S MeHg Oxidation SRB Hg II Plants Trophic Transfer Hg II Anaerobic Transformations in Aquatic Ecosystems Nitrate in surface waters inhibits Hg methylation since denitrifiers can out compete Fe- and Sulfate-reducing bacteria for energy (reduced carbon compounds). A Tale of Two Elements

7. Methylation Theoretical Considerations Bioavailability of Hg II in Natural Ecosystems RSH + Hg(OH) 2 + 2 H +  RS-Hg (unavailble) H 2 S + RS-Hg  Hg(SH) 2 Sulfate-reducing bacteria control this reaction by producing H2S. SRB also have the right enzymes to methylate. Fe reducers do as well, but may depend on SRB to make Hg II bioavailable.

8. Clear Pond, Kickapoo State Park Profile (August 2010) MeHg Analysis at UIUC by W.Y. Chen

9. Landscape-Scale Effects of Natural Anaerobic Ecosystems on Mercury Cycling Brigham et al. (2009)

10. Constructed Anaerobic Ecosystems

11. Denitrifying Bioreactors: Constructed Anaerobic Ecosystems Pioneered by Richard Cooke, ABE-UIUC. Designed to reduce nitrate export from tile-drained fields. Economical Small footprint Wood chips are inexpensive Minimal cost of water control structures (<$10k) Little time required to operate/maintain Field tests show their high efficiency at NO 3 - removal. Denitrifying Bioreactors

12. Monticello Bioreactor Site Denitrifying Bioreactors

14. Capacity Control Structure Woodchips Diversion Structure Second Generation Bioreactors Denitrifying Bioreactors

15. Andrus et al. (2010) Denitrifying Bioreactors

16. Capacity control structure Up to soil surface Side View Trench bottom 1’ below tile invert 5’ section of non-perforated tile Length dependent on treatment area Diversion structure Top View 5’ Soil backfill 10’ Wide Denitrifying Bioreactors

18. Biogeochemistry Denitrifying Bioreactors Abrus et al.

19. Bioreactor Methylmercury Study Objective 1.Determine whether denitrifying bioreactors produce methylmercury due to anoxic environments formed within bioreactor. 2.Determine total Hg export flux. 3.Compare to natural levels in environment.

20. Bioreactor Methylmercury Study Design Synoptic sampling design – Collect samples after storm events and other periods when tiles are flowing – Inlet and outlet sampled simultaneously – Sampled periodically from summer 2008-June 2009 – Sampled again in summer 2010. – Preserved by filtration/acidification or freezing. Analyze – Dissolved Methylmercury (MeHg) – Dissolved organic carbon – Sulfate, nitrate, chloride – Only a limited number of samples have been analyzed to date.

21. Sampling Bioreactors Inlet Samples Outlet Samples

22. UIUC Method for Hg Speciation Analysis Shade and Hudson (2005) Environmental Science and Technology Shade, Hudson, et al. patent (circa 2007) Vermillion and Hudson (2007). Analytical and Bioanalytical Chemistry. Preparation method for water samples.

23. pH-Modulated Thiol-Thione Switch After Laws (1970) + Thiol Resin -S-H + Thiol Resin -S- -S-H + pH<2: Desorption Favored pH > 3: Adsorption Favored H+H+ H H University of Illinois at Urbana-Champaign

24. Thiourea Complexes of Hg 2+ and CH 3 Hg + + 2+ MeHg + Hg 2+ University of Illinois at Urbana-Champaign

25. Mercury-thiourea complex ion chromatography Shade and Hudson, ES&T 2005 University of Illinois at Urbana-Champaign

26. Dissolved methylmercury: Thiourea-catalyzed solid phase extraction Vermillion and Hudson Analytical and Bioanalytical Chemistry (2007) Brian Vermillion University of Illinois at Urbana-Champaign

27. Validation of Sediment and Biota Sample Preparations Biota: Leaching of tissues in acidic TU (Shade, ES&T 2008) Sediments: H 2 SO 4 +KBr Digestion/Toluene Extraction (Vermillion, Shade, and Hudson, in prep)

28. Comparison of Methods for Analysis of Dissolved MeHg Ultraclean sample collection Sample preservation Extraction from Sample Matrix Pre- concentration/ Loading Chromatographic Separation of Hg species Sensitive detection Distillation________ TU-Catalyzed SPE Ethylation /Purge & Trap __________ Elute SPE /Online Trap Gas Chromatography _____________ Ion Chromatography AFS_________AFS University of Illinois at Urbana-Champaign

29. Intercomparison with Distillation/Ethylation ICP-MS Trent University (Hintelmann)

30. Intercomparison with USGS Wisconsin District Mercury Lab (Krabbenhoft)

31. Bioreactor Study Results

32. Dissolved MeHg in Bioreactor Inlets -Eight non-detects -Six samples contained detectable MeHg - Maximum: 0.16 ng/L - Maximum: 0.16 ng/L - Average:0.09 ng/L - Average:0.09 ng/L

33. Dissolved MeHg in Bioreactor Outlets

34. Farm Progress City Bioreactor

35. Nitrate Removal

37. “Typical” Hg Levels Groundwater Total Hg is very low (<0.5 ng/L) MeHg is often non-detectable. Surface Waters Total Hg is usually 1-5 ng/L in unpolluted systems MeHg is usually <0.5 ng/L in unpolluted systems

38. Possible Relationship between Methylmercury Production to Sulfate Consumption in Bioreactors

39. Dissolved Organic Carbon Dissolved organic carbon is known to be a carrier of mercury and methylmercury. DOC in inlets: Geometric mean of 1.6 mg-C/L DOC in outlets: Geometric mean of 16 mg-C/L Exhibits seasonality

40. Summary of Results to Date Levels in bioreactor discharge are much higher than in tile drain water entering bioreactors. Methylmercury is clearly produced in bioreactors. Some combination of increase in DOC and decrease in sulfate may be responsible. Levels in bioreactor discharge are much higher than typical surface water values (0.1-0.3 ng/L).

41. What is source of Hg? Tile water: Groundwater is typically very low except when preferential flow occurs Could accumulate on wood chips during high flow (aerobic conditions) and be released under high flow conditions Wood chips: Trees accumulate Hg from air and soil water in the wood

42. Methylation Theoretical Considerations Bioavailability of Hg II in Natural Ecosystems RSH + Hg(OH) 2 + 2 H +  RS-Hg (unavailble) H 2 S + RS-Hg  Hg(SH) 2 Sulfate-reducing bacteria control this reaction by producing H2S. SRB also have the right enzymes to methylate. Fe reducers do as well, but may depend on SRB to make Hg II bioavailable.

43. Questions What controls the extent of methylation? – Amount of sulfate in tile drainage? – Extent of anoxia? – Flow rate? Can reactors be designed to minimize MeHg formation?

44. Capacity control structure Up to soil surface Side View Trench bottom 1’ below tile invert 5’ section of non-perforated tile Length dependent on treatment area Diversion structure Top View 5’ Soil backfill 10’ Wide Trench bottom at tile invert Denitrifying Bioreactors

45. Implications MeHg is produced in some bioreactors. Effect on MeHg fluxes in watersheds needs to be considered An apparent trade-off: Reducing nitrate levels to make water drinkable may make the waters downstream unfishable.

46. Implications (cont.) Reduction of MeHg production likely can be attained by adjusting design: Eliminate ponding zones Use of low-Hg wood (hypothesis)