microbial diversity in drain hole waters of the henderson mine
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Microbial Diversity in Drain Hole Waters of the Henderson Mine. John R. Spear Jason Sahl Division of Environmental Science and Engineering Colorado School of Mines, Golden, CO [email protected]. $125 x 10 6. $125 x 10 6. Amount spent in the U.S. on marshmallows in 2005. What Is Life?. - PowerPoint PPT PresentationTRANSCRIPT
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Microbial Diversity in Drain Hole Waters of the Henderson Mine
John R. SpearJason Sahl
Division of Environmental Science and EngineeringColorado School of Mines, Golden, CO
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$125 x 106
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$125 x 106
Amount spent in the U.S. on marshmallows in 2005
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What Is Life?
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Euryarchaeota
Crenarchaeota
A “Map” of LifeEuryarchaeota
Crenarchaeota
N.R. Pace, Science, 1997
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Molecular MethodEnv. Sample
Bulk DNA/RNA
Extraction
CommunityrRNA/rDNA
rRNA/rDNAClones
PCR
Cloning
rRNA/rDNA Sequences
Sequencing
PhylogeneticTrees
Databases
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7150’ DH-3 Data
10%
14%
29%14%
3%
3%
17%
10%
Fungus; AscomycotaGreen Non-Sulfur or ChlorofexiDelta-ProteobacteriumBeta-ProteobacteriumAlpha-ProteobacteriumSpirocheteActinobacteriumFirmicutes; Clostridium
Fungus
Sulfate Reducing Bacteria
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7025’ DH-1 Data
Fungus
Sulfate Reducing Bacteria
73%
1%4%5%
16%
1%
B-Proteobacteria
Nitrosomonas
Acidobacteria
Henderson Ungrouped#3Bacteroidetes
Fungi
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7025’ DH-1-Post Packer Data
Fungus
Sulfate Reducing Bacteria
6%4%
1%
60%
14%
12%
1%1%
1%Henderson Ungrouped#1Henderson Ungrouped#3B-Proteobacteria
Chloroflexi
Chlorbi
Firmicutes
Nitrospira
OS-K
NBO1
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7025’ DH-4-Post Packer Data
Fungus
Sulfate Reducing Bacteria
3% 1%5%
7%
1%
39%
44%
Henderson Ungrouped#1Gamma-Proteo
Delta-Proteo
Acidobacteria
Chloroflexi
Chlorobi
Crenarchaeota
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DH Chemistry7025-D1 7025-P1 7025-D4 7025-P4
O2 (mg/L) 0.26 0.14 3.8 0.04Temp (oC) 38 40 35 35
pH 5.9 5.8 6.3 6DIC (mM) 29 16 3.2 20DOC (mM) 0.05 0.03 0.05 0.03
Total N (mM) 0.05 0.05 0.17 0.17SO4
2- (mM) 4.3 4 10 9.8NO3
- (mM) 0.03 0.017 0.086 0.047NO2
- (mM) 0.05 0.027 0.048 0.046
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HM
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Conclusions
Potential for 3 new groups (phyla) of Bacteria!!Something is happening with N!!Community shift pre- verses packed- drill
holes.Surprising diversity for a subsurface.Low abundance of both Archaea and Eucarya!Publish?
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Thanks To:
My Start-up money….Team Bio--Tom K., Jeff M., Alexis T.,
Betsy S., Jeff W., Kevin M.HUSEP FolksHenderson Mine and Chip deWolfe
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“If it wasn’t for the optimist,the pessimist would never know….
how happy he wasn’t.”
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Henderson, New Drill Core
Plan, to drill a core into the HUSEP facility area
For some portion of that:– Take a segment for biological consideration
• Sterile? BEADS + Tracer• Core collection--on site• Core sample sections to Socorro, CSM, CU
– Continue drilling to depth
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Henderson Mine, 2 Nov 05
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Molecular MethodEnv. SampleContact Slide
Bulk DNA/RNA
Extraction
CommunityrRNA/rDNA
rRNA/rDNAClones
PCR
Cloning
= Method = Result
rRNA/rDNASequences / Databases
Nucleic AcidProbes
Sequencing
ProbeDesign
Whole-CellHybridization
(FISH)
PhylogeneticTrees
Nucleic Acid
Hybridization
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GGCC
TACG
CCGG TACGGGCC ATGC
Polymerase Chain Reaction
CCGG TACG
GGCC ATGC
GGCC ATGC
CCGG TACG
GGCC ATGC
TACG
CCGG TACGGGCC
GGCC ATGC
TACG
CCGG TACG
GGCC
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DH-2, DH-3 + ???? To Do….
Water
Water
= 1.5’ PVC Pipe Inserted with Glass and/or Fe Growth Slides
Anoxic H2Oout
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DH-1 Water Chemistry + CulturesSpecies Concentration Si4+ 0.05 mMCa2+ 8.7 mMMg2+ 1.3 mMAl3+ 4.8 mMNa+ 7.9 mMSr2+ 0.04 mMK+ 1.0 mMFeT 1.5 mMMn2+ 21 mMF- 12 mMCl- 0.15 mMNO2- Not detectedNO3- 1.0 mMBr- Not detectedPO43- Not detectedSO42- 48 mM
DH-1 at 7150’, pH 5.8
Culture efforts:Swanner & Templeton, CU:
DH-1 3 (Fe + Mn)DH-3 3 (Mn)DH-4 2 (Fe + Mn)