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Davis Mine Project

Davis Mine Project. By: Jason Jean and Janice Wing. Davis mine. Drainage below tailings pile. Tailings pile. Drainage below tailings pile. Drainage below another tailings pile. Sample of pyrite at Davis Mine. Stream confluence upstream from A4. Stream confluence up close.

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Davis Mine Project

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  1. Davis Mine Project By: Jason Jean and Janice Wing

  2. Davis mine

  3. Drainage below tailings pile

  4. Tailings pile

  5. Drainage below tailings pile

  6. Drainage below another tailings pile

  7. Sample of pyrite at Davis Mine

  8. Stream confluence upstream from A4

  9. Stream confluence up close

  10. Presentation Outline • Reasons for our research project • Methods • Findings • Obstacles • Research continuation • Impact on us

  11. Reasons for our research project • Personal • Professional • Enrichment media

  12. Methods • Sterile, anaerobic technique • pH adjustment • Inoculation • Incubation • Observations and Microscopy

  13. Acquiring mine water from well

  14. Collection bottles for Davis Mine groundwater

  15. Measuring well water depth

  16. Time to think

  17. Sterilize bottles in the acid wash

  18. Preparing enrichment media under anoxic conditions.

  19. pH adjustment

  20. Sterilize Postgate and Pfennig media before inoculation in the autoclave. • Davis Mine Well media was not sterilized in the autoclave before inoculation.

  21. Sediment from Well 1, 3, 4 were used

  22. Opening sediment jars and inoculating sediment into media bottles under anoxic conditions

  23. Incubation and observation

  24. Storage and incubation of transfer bottles

  25. Findings • Sulfur-reducing bacteria grew in Postgate pH 7.0 • Sulfur-reducing bacteria grew in Davis Mine Well 1 Groundwater media at pH 7.0 • Thread-like filaments were visible in sample jars that developed a black precipitate, but in no others. • Desulfobulbus was successfully cultivated in Postgate media but was not successful in Pfennig. • Orange film developed in Pfennig enrichment bottles.

  26. Enrichment bottles with Postgate’s medium at pH 3.0 4.0 5.0 7.0

  27. Enrichment bottles with Pfennig’s medium at pH 3.0 4.0 5.0 7.0

  28. Davis Mine Well 1 media at pH 3.0 4.0 5.0 7.0

  29. Davis Mine Well 3 media at pH 3.0 4.0 5.0 7.0

  30. Davis Mine Well 4 media at pH 3.0 4.0 5.0 7.0

  31. Black precipitate formed in Postgate media of pH 7.0 inoculated with Desulfobulbus

  32. Black precipitate formed in transfers of Postgate media at pH 7.0

  33. Black precipitate formed in transfers of Davis Mine Well 1 media at pH 7.0

  34. Microscopy using a live/dead bio-stain with this fluorescent microscope. Pictures were taken.

  35. Microscopy of Postgate pH 3.0 sample inoculated with Well 1 sediment

  36. Microscopy of Postgate pH 4.0 inoculated with Well 4 sediment

  37. Postgate at pH 5.0, sediment from Well 4 Cluster

  38. Postgate pH 7.0 sediment from Well 4 Thread-like filament

  39. Postgate pH 7.0 sediment from Well 4 Dark spots of pyrite forming

  40. What is the orange film and sediment? (Seen in all Pfennig media, here the sample is pH 3.0)

  41. Obstacles • Time • Equipment • Anaerobic technique is rigorous

  42. Presence of oxygen

  43. Contamination by a fungus

  44. Tough going

  45. Research continuation • Dilution transfers and DNA extraction • Continued incubation • Postgate media with a variety of donors • DM Well media with a variety of donors • Mercy • Iron-reducers

  46. Impact on us • Better understanding of long-range research • Lab and project research experience • Integrate into classroom curriculum aligned with state frameworks • Biology • Environmental Science

  47. Questions ??

  48. The End

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