microbial diversity in drain hole waters of the henderson mine
DESCRIPTION
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
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
$125 x 106
$125 x 106
Amount spent in the U.S. on marshmallows in 2005
What Is Life?
You are here
Euryarchaeota
Crenarchaeota
A “Map” of LifeEuryarchaeota
Crenarchaeota
N.R. Pace, Science, 1997
Molecular MethodEnv. Sample
Bulk DNA/RNA
Extraction
CommunityrRNA/rDNA
rRNA/rDNAClones
PCR
Cloning
rRNA/rDNA Sequences
Sequencing
PhylogeneticTrees
Databases
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
7025’ DH-1 Data
Fungus
Sulfate Reducing Bacteria
73%
1%4%5%
16%
1%
B-Proteobacteria
Nitrosomonas
Acidobacteria
Henderson Ungrouped#3Bacteroidetes
Fungi
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
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
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
HM
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?
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
“If it wasn’t for the optimist,the pessimist would never know….
how happy he wasn’t.”
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
Henderson Mine, 2 Nov 05
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
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
DH-2, DH-3 + ???? To Do….
Water
Water
= 1.5’ PVC Pipe Inserted with Glass and/or Fe Growth Slides
Anoxic H2Oout
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)