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

$125 x 10 6

Amount spent in the U.S. on marshmallows in 2005

What Is Life?

You are here Euryarchaeota Crenarchaeota A “Map” of Life Euryarchaeota Crenarchaeota N.R. Pace, Science, 1997

Molecular Method Env. Sample Bulk DNA/RNA Extraction Community rRNA/rDNA Clones PCR Cloning rRNA/rDNA Sequences Sequencing Phylogenetic Trees Databases

7150’ DH-3 Data Fungus Sulfate Reducing Bacteria

7025’ DH-1 Data Fungus Sulfate Reducing Bacteria

7025’ DH-1-Post Packer Data Fungus Sulfate Reducing Bacteria

7025’ DH-4-Post Packer Data Fungus Sulfate Reducing Bacteria

DH Chemistry

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 Folks Henderson 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 Method Env. Sample Contact Slide Bulk DNA/RNA Extraction Community rRNA/rDNA Clones PCR Cloning = Method= Result rRNA/rDNA Sequences / Databases Nucleic Acid Probes Sequencing Probe Design Whole-Cell Hybridization (FISH) Phylogenetic Trees Nucleic Acid Hybridization

DH-2, DH-3 + ???? To Do…. Water = 1.5’ PVC Pipe Inserted with Glass and/or Fe Growth Slides Anoxic H 2 O out

DH-1 Water Chemistry + Cultures SpeciesConcentration Si mM Ca2+8.7 mM Mg2+1.3 mM Al3+4.8 mM Na+7.9 mM Sr mM K+1.0 mM FeT1.5 mM Mn2+21 mM F-12 mM Cl-0.15 mM NO2-Not detected NO3-1.0 mM Br-Not detected PO43-Not detected SO42-48 mM DH-1 at 7150’, pH 5.8 Culture efforts: Swanner & Templeton, CU: DH-13 (Fe + Mn) DH-33 (Mn) DH-42 (Fe + Mn)