1. By: Jason Jean and Janice Wing
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 atpH

27. Enrichment bottles with Pfennig’s medium atpH

28. Davis Mine Well 1 media atpH

29. Davis Mine Well 3 media atpH

30. Davis Mine Well 4 media atpH

31. Black precipitate formed in Postgate media of pH 7
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