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Princeton University, University of the Free State, South Africa,

From bacterial finches to bactivorous meiofauna : Microbial biogeography of the deep crustal biosphere. T.C. Onstott 1 , C. Chehoud 1 , E. van Heerden 2 , D. Litthauer 2 , A. Jugdave 2 , K. Albertyn 2 , T. Kieft 3 , G. Borgonie 4 , A. García-Moyano 5 and D. Chivian 6.

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Princeton University, University of the Free State, South Africa,

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  1. From bacterial finches to bactivorousmeiofauna: Microbial biogeography of the deep crustal biosphere T.C. Onstott1, C. Chehoud1, E. van Heerden2, D. Litthauer2, A. Jugdave2, K. Albertyn2, T. Kieft3,G. Borgonie4, A. García-Moyano5 and D. Chivian6 Princeton University, University of the Free State, South Africa, New Mexico Institute of Mining and Technology, University of Ghent, Belgium, University of Bergen, Bergen, Norway, Lawrence Berkeley National Lab, USA (DCChivian@lbl.gov)

  2. Evander How long can deep, sequestered microbial communities survive?

  3. Northam Platinum Mine – Bushveld Igneous Complex Age: 2054 Ma Cr, PGE, V Temp: 42°C – 53°C pH: 8 – 11 Barnes et al., 2004; Scoateset al., 2008; Wangeret al., 2008

  4. 16S rRNA gene sequence analysis • 1,916 sequences deposited in NCBI from 58 sites and 84 sites+time points. • 983 OTUs identified at 97% identity • 718 OTUs occur only once • 16S rRNA gene sequences classified as contaminants because • Present in 10 sites of mining water, mine air and acid mine water • Present in same OTU as found in mining water, mine air and acid mine water • Guilt by association – closely associated with contaminant OTUs on phylogenetic tree. • 279 OTUs labeled as contaminants • %contaminated sequences varied from 0 to 97% with 8 sites+time points with ≥50% contaminants were removed from canonical analysis. • Taxonomic identification to the family level with 80% confidence.

  5. 279 Contaminant OTUs out of 982 total OTUs. Guilt by association

  6. Family Richness in 8 Mines 1

  7. Mining Water from Kloof, Driefontein, Mponeng and Evander Beatrix (Lin et al. 2006) Observed OTU’s Evander 2 and 8 shaft combined Kloof, Mponeng and Driefontein combined Sampled Sequences

  8. Mine Archaeal Family Compositions

  9. Mine Bacterial Family Composition

  10. Mine Bacterial Family Composition

  11. biofilms rock cartridges borehole water Species number Depth (kmbls.)

  12. Eveness Depth (kmbls.)

  13. Only 50 OTUs occur at more than one site and only 3 OTUs are distributed across the basin. Sorensen Similarity Index Horizontal Distance (km)

  14. Tree Showing the Distances between the OTUs

  15. 3.3 km 2.0 km 3.2 km 1.7 km 2.8 km 2.7 km 1 km 1.5 km

  16. Conclusion The lack of overlap in OTUs between 37 fractures sampled to date suggests an “desert oases” biogeographical pattern that could encompass ~8,000 OTUs within the Witwatersrand Basin. A combination of pyrosequencing and rock filters may reveal a greater diversity – hopefully with the help of the CoDL. Singe cell genomes of the “finches” will enable a more direct correlation of genome structure, environment and geographic dispersion.

  17. EstavanHeerden Tom Kieft Derek Litthauer Duane Moser GaetanBorgonie

  18. Acknowledgements NSF - Geobiologyand Low-Temperature Geochemistry Program Sloan Foundation through the Deep Carbon Observatory

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