Davis Mine Project

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Davis Mine Project

• By Jason Jean
• and Janice Wing

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Davis mine
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Drainage below tailings pile
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Tailings pile
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Drainage below tailings pile
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Drainage below another tailings pile
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Sample of pyrite at Davis Mine
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Stream confluence upstream from A4
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Stream confluence up close
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• Presentation Outline
• Reasons for our research project
• Methods
• Findings
• Obstacles
• Research continuation
• Impact on us

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

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

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Acquiring mine water from well
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Collection bottles for Davis Mine groundwater
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Measuring well water depth
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Time to think
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Sterilize bottles in the acid wash
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Preparing enrichment media under anoxic
conditions.
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pH adjustment
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• Sterilize Postgate and Pfennig media before
  inoculation in the autoclave.
• Davis Mine Well media was not sterilized in the
  autoclave before inoculation.

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Sediment from Well 1, 3, 4 were used
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Opening sediment jars and inoculating sediment
into media bottles under anoxic conditions
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Incubation and observation
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Storage and incubation of transfer bottles
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• 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.

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Enrichment bottles with Postgates medium at pH
3.0 4.0 5.0 7.0
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Enrichment bottles with Pfennigs medium at pH
3.0 4.0 5.0 7.0
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Davis Mine Well 1 media at pH 3.0 4.0 5.0
7.0
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Davis Mine Well 3 media at pH 3.0 4.0
5.0 7.0
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Davis Mine Well 4 media at pH 3.0 4.0
5.0 7.0
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Black precipitate formed in Postgate media of pH
7.0 inoculated with Desulfobulbus
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Black precipitate formed in transfers of Postgate
media at pH 7.0
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Black precipitate formed in transfers of Davis
Mine Well 1 media at pH 7.0
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• Microscopy using a live/dead bio-stain with this
  fluorescent microscope. Pictures were taken.

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Microscopy of Postgate pH 3.0 sample inoculated
with Well 1 sediment
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Microscopy of Postgate pH 4.0 inoculated with
Well 4 sediment
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Postgate at pH 5.0, sediment from Well 4
Cluster
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Postgate pH 7.0 sediment from Well 4
Thread-like filament
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Postgate pH 7.0 sediment from Well 4
Dark spots of pyrite forming
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What is the orange film and sediment? (Seen in
all Pfennig media, here the sample is pH 3.0)
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• Obstacles
• Time
• Equipment
• Anaerobic technique is rigorous

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Presence of oxygen
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Contamination by a fungus
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Tough going
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• 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

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• 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

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Questions ??
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The End