Dr. John G. Woiwode, Principal Investigator AquaMatrix International, Inc.

1. The Use of Intensively Cultured Fish and Halophytic Plants for the Bioremediation of Coal Bed Methane (CBM) Discharge Waters Dr. John G. Woiwode, Principal Investigator AquaMatrix International, Inc.

2. Grant Writing • The Story • The Plot • The Bottleneck • The Hypothesis • The Experimental Design • The Conclusions

3. Phase 1 Feasibility of an idea, a widget Vs the Feasibility of a Concept Phase 2 Development of a Commercial Prototype SBIR

4. Aquaculture Industry Participation • Essential that the fish are not perceived as tainted • Essential that there are no public health concerns

5. Bioaccumulation • FDA regulates As, Hg, Pb, Ni, Cr and Cd • B, Ba, Fe and Mn also high in solution • Hydrocarbons

6. Experimental Design • 90 day culture • RBT • 3 Aquaria Test, 3 Control • Assay anterior epaxial muscle

7. Results • No Bioaccumulation of: As, Cd, Cr, Pb, Hg, Ni • No Bioaccumulation of Fe, Mn, B • “Significant” Bioaccumulation of Ba • No Bioaccumulation of 200 organic hydrocarbons • Growth Commercially Acceptable

8. The Direct Use of Coal Bed Methane Discharge Water for Intensive Aquaculture Applications U.S. Department of Agriculture

9. Commercial Prototype Development • Degas, regas issues

10. Commercial Fish Production • Modular • Carbonates, pH; Recirculating • Degas/regas; Cations • Density and Growth

11. CBM and Fish Discharge • Use of Fish Manures • CBM Discharge is Contentious • CBM Discharge as Irrigant

12. Halophytes • Aggressively Consume Salts • Commercial Species: Oil Seed Fiber Fodder Forage

13. The Use of Halophytic Plants for the Bioremediation of Coal Bed Methane Discharge Waters National Science Foundation

14. Experimental Design • Three Plant Species: Four Wing Salt Bush, Malt Barley and Crested Wheat grass • Four Water Treatments: CBM water, CBM water with inorganic fertilizer, CBM water with fish manures, and control of spring water • Four repetitions

15. Results

16. Results The plant tissue contained: • An order of magnitude higher Na levels when irrigated with CBM water. • 2-300% greater growth when irrigated with CBM water and fish manure. • yet an additional 200-300% more Na/kg when irrigated with CBM with fish manures.

17. Results • Total sodium removal by four wing salt bush grown with CBM water and fish manures was 18.3 kg/acre, and 38.5 kg/acre with malting barley. • Soil remained below the target SAR of 10 for all CBM irrigated treatments.

18. Four Wing Salt Bush • Sodium uptake was significant into the leaves • Use of Fish Manures enhanced the uptake of Sodium

19. CBM Discharge Implications • Soil structure appears to improve • Discharge permitting may be affected, as the classification could be changed from “pollutant” to “agricultural discharge” after going through fish • Divalent Cations, Carbonates • Sodium uptake from soil and water by Halophytes, further enhanced by Fish Manures

20. Implications • Halophytes irrigated with CBM waters and fish manures uptake significant Sodium from the water and soil. • Malt Barley irrigated with CBM water and fish manures could produce a high value commercial crop. • Landowners would welcome a beneficial forage species grown on their property. Forage animals would be able to derive significant nutrition from select plants irrigated with CBM discharge water. • Noxious weeds encroaching on discharge areas would be displaced by organized agriculture of halophytic plants. • Surface soils would ultimately contain less salt.

21. The Use of Halophytes and Fish Production for the Bioremediation of Coal Bed Methane Discharge Waters • Field trials of nine plant species irrigated with CBM discharge, combined with manures from a commercial fish farm. • The larger vision: The Agronomic Model of Fish and Halophytes