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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
Presentation Outline
•Reasons for our research project
•Methods
•Findings
•Obstacles
•Research continuation
•Impact on us
Reasons for our research project
•Personal
•Professional
•Enrichment media
Methods
• Sterile, anaerobic technique
• pH adjustment
• Inoculation
• Incubation
• Observations and Microscopy
Acquiring mine water from well
Collection bottles for Davis Mine groundwater
Measuring well water depth
Time to think
Sterilize bottles in the acid wash
Preparing enrichment media under anoxic conditions.
pH adjustment
Sterilize Postgate and Pfennig media before inoculation in the autoclave. Davis Mine Well media was not sterilized in the autoclave before inoculation.
Sediment from Well 1, 3, 4 were used
Opening sediment jars and inoculating sediment into media bottles under anoxic conditions
Incubation and observation
Storage and incubation of transfer bottles
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.
Enrichment bottles with Postgate’s medium at
pH 3.0 4.0 5.0 7.0
Enrichment bottles with Pfennig’s medium at
pH 3.0 4.0 5.0 7.0
Davis Mine Well 1 media at
pH 3.0 4.0 5.0 7.0
Davis Mine Well 3 media at
pH 3.0 4.0 5.0 7.0
Davis Mine Well 4 media at
pH 3.0 4.0 5.0 7.0
Black precipitate formed in Postgate media of pH 7.0 inoculated with Desulfobulbus
Black precipitate formed in transfers of Postgate media at pH 7.0
Black precipitate formed in transfers of Davis Mine Well 1 media at pH 7.0
Microscopy using a live/dead bio-stain with this fluorescent microscope. Pictures were taken.
Microscopy of Postgate pH 3.0 sample inoculated with Well 1 sediment
Microscopy of Postgate pH 4.0 inoculated with Well 4 sediment
Postgate at pH 5.0, sediment from Well 4
Cluster
Postgate pH 7.0 sediment from Well 4
Thread-like filament
Postgate pH 7.0 sediment from Well 4
Dark spots of pyrite forming
What is the orange film and sediment?
(Seen in all Pfennig media, here the sample is pH 3.0)
Obstacles
• Time
• Equipment
• Anaerobic technique is rigorous
Presence of oxygen
Contamination by a fungus
Tough going
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
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
Questions ??
The End