Searching for Microbes and DNA in Ancient Halite

  • Russell H. VreelandEmail author


David Hume (1711–1767) is credited with coining the phrase “A wise man, therefore, proportions his belief to the evidence.” This was later brought to a more modern understanding by Marcello Truzzi as “An extraordinary claim requires extraordinary proof” (Truzzi 1978). Certainly such an admonition should be considered by any and all scientists attempting to carry out research on ancient living microbes or DNA. Such supporting evidence should include several factors with publications and reports often seeming to be more about the evidence than about the discovery. The key bits of supporting information described here range from the overall geological pedigree of the formation and the individual sample, the sterilization assurance levels and even the design of the laboratory in which the work occurred. This chapter outlines how to establish these various types of evidence in order to support claims that any isolate or DNA sequence is as old as the crystal in which it was found. Adherence to these ideas will never make claims of ancient life immediately accepted. However, experience has shown that having these various bits of information on hand will at least make the debate more likely to have a positive outcome.


Sterility Assurance Live Microbes Ancient Material Ancient Salt Microbal Ance 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Bobst AL, Lowenstein TK, Jordan TE, Godfrey LV, Hein MC, Ku TL, Luo S (2001) A 106 kapaleoclimate record from the Salar de Atacama, northern Chile. Palaeogeo Palaeoclimatol Palaeoecol 173:21–42CrossRefGoogle Scholar
  2. Cano RJ, Borucki MK (1995) Revival and identification of bacterial spores in 25–40 million year old Dominican Amber. Science 268:1060–1064CrossRefGoogle Scholar
  3. Chernikoff S (1999) Geology, 2nd edn. Houghton Mifflin Co., New YorkGoogle Scholar
  4. Dombrowski H (1963) Bacteria from Paleozoic salt deposits. Annu NY Acad Sci 108:453–460CrossRefGoogle Scholar
  5. Dombrowski H (1966) Geological problems in the question of living bacteria in Paleozoic salt deposits. Second Symposium on Salt 1:215–220Google Scholar
  6. Farrell MF, Turner HG (1932) Bacteria in anthracite coal. J Bacteriol 23:155–162PubMedPubMedCentralGoogle Scholar
  7. Fish SA, Shepherd TJ, McGenity TJ, Grant WD (2002) Recovery of 16S ribosomal RNA gene fragments from ancient halite. Nature 417:432–436CrossRefGoogle Scholar
  8. Grant WD, Gemmell RT, McGenity TJ (1998) Halobacteria: the evidence for longevity. Extremophiles 2:279–287CrossRefGoogle Scholar
  9. Graur D, Pupko T (2001) The Permian bacterium that isn’t. Mol Biol Evol 18:1143–1146CrossRefGoogle Scholar
  10. Griffith JD, Wilcox S, Powers DW, Nelson R, Baxter B (2008) Discovery of abundant cellulose microfibers encased in 250 Ma Permian halite: a macromolecular target in the search for life on other planets. Astrobiology 8:215–228CrossRefGoogle Scholar
  11. Hazen RM, Roedder E (2001) How old are bacteria from the Permian age? Nature 411:155CrossRefGoogle Scholar
  12. Holt RM, Powers DW (1990) Geologic mapping of the air intake shaft at the Waste Isolation Pilot Plant. U.S. Department of Energy, Carslbad, NM, DOE/WIPP 90-051Google Scholar
  13. Kiminek G, Bada JL, Pogliano K, Ward JF (2003) Radiation-dependent limit for the viability of bacterial spores in halite fluid inclusions and on Mars. Radiat Res 159:722–729Google Scholar
  14. Lipman CB (1931) Living microorganisms in ancient rocks. J Bacteriol 22:183–198PubMedPubMedCentralGoogle Scholar
  15. Lipman CB (1937) Bacteria in coal. J Bacteriol 34:483–488PubMedPubMedCentralGoogle Scholar
  16. Lowenstein TK, Li J, Brown C, Roberts SM, Ku Tl, Luo S, Yang W (1999) 200 k.y. paleoclimate record from Death Valley salt core. Geology 27:3–6CrossRefGoogle Scholar
  17. Mileikowsky C, Cucinotta FA, Wilson JA, Gladman B, Lindegren L, Melosh J, Rickman H, Valtonen M, Zheng JQ (2000) Natural transfer of viable microbes in space 1. from Mars to Earth and Earth to Mars. Icarus 145:391–427CrossRefGoogle Scholar
  18. Mormile MR, Biesen MA, Gutierrez MC, Ventosa A, Pavolvich JB, Onstott TC, Frederickson JK (2003) Isolation of Halobacterium salinarum retrieved directly from halite brine inclusions. Environ Microbiol 5:1094–1102CrossRefGoogle Scholar
  19. Nicastro AJ, Vreeland RH, Rosenzweig WD (2002) Limits imposed by ionizing radiation on the long term survival of trapped bacterial spores. J Radiation Biology 78:891–901CrossRefGoogle Scholar
  20. Nickle DC, Learn GH, Rain MW, Mullins JI, Mittler JE (2002) Curiously modern DNA for a “250 million-year-old” bacterium. J Mol Evol 54:134–137CrossRefGoogle Scholar
  21. Norton CF, Grant WD (1988) Survival of halobacteria within fluid inclusions in salt crystals. J Gen Microbiol 134:1365–1373Google Scholar
  22. Norton CF, McGenity TJ, Grant WD (1993) Archaealhalophilies (halobacteria) from two British salt mines. J Gen Microbiol 139:1077–1081CrossRefGoogle Scholar
  23. Park JS, Cho BC, Lowenstein TK, Timofeeff MN, Rosenzweig WD, Vreeland RH (2009) Ancient primary salts confirm long term DNA survival and show aspects of microbial evolution and ecology. Geobiology 7:515–523CrossRefGoogle Scholar
  24. Powers DW, Vreeland RH, Rosenzweig WD (2001) Biogeology—How old are bacteria from the Permian age?—Reply Nature 411:155–156CrossRefGoogle Scholar
  25. Radax C, Gruber C, Stan-Lotter H (2001) Novel haloarchaeal 16S rRNA gene sequences from Alpine Permo-Triassic rock salt. Extremophiles 5:221–228CrossRefGoogle Scholar
  26. Reiser R, Tasch P (1960) Investigation of the viability of osmophile bacteria of great geological age. Trans Kans Acad Sci 63:31–34CrossRefGoogle Scholar
  27. Rosenzweig WD, Woish J, Petersen J, Vreeland RH (2000) Development of a protocol to retrieve microorganisms from ancient salt crystals. Geomicrobiology 17:185–192CrossRefGoogle Scholar
  28. Satterfield CL, Lowenstein TK, Vreeland RH, Rosenzweig WD, Powers DW (2005) New evidence for 250Ma age of halophilic bacterium from a Permian salt crystal. Geology 33:265–268CrossRefGoogle Scholar
  29. Schubert BA, Lowenstein TK, Timofeeff MN (2009a) Microscopic identification of prokaryotes in modern and ancient halite, Saline Valley and Death Valley CA. Astrobiology 9:467–482CrossRefGoogle Scholar
  30. Schubert BA, Lowenstein TK, Timofeeff MN, Parker MA (2009b) How do prokaryotes survive in fluid inclusions in halite for 30 k.y.? Geology 37:1059–1062CrossRefGoogle Scholar
  31. Stan-Lotter H, McGenity TJ, Legat A, Denner EBM, Glaser K, Stetter KO, Wanner G (1999) Very similar strains of Halococcus salifodinae are found in geographically separated Permo-Triassic salt deposits. Microbiology 145:3565–3574CrossRefGoogle Scholar
  32. Stan-Lotter H, Radax C, Gruber C, McGenity TJ, Legat A, Wanner G, Denner EBM (2000) The distribution of viable microorganisms in Permo-Triassic rock salt. In: Geertman RM (ed) SALT 2000. 8th World Salt Symposium Amsterdam. Elsevier, Amsterdam, The NetherlandsGoogle Scholar
  33. Stan-Lotter H, Radax C, McGenity TJ, Legat A, Pfaffenhuemer M, Wieland H, Gruber C, Denner EBM (2001) From intraterrestrials to extraterrestrials—viable haloarchaea in ancient salt deposits. In: Ventosa A (ed) HalophilesSpringer, pp 89–102Google Scholar
  34. Tadros ME, Tucker MD (2003) Formulations for neutralization of chemical and biological toxants. US Patent 6:566–574 B1Google Scholar
  35. Tasch P (1960) Paleoecological observations of the Wellington salt (Hutchinson member). Trans Kans Acad Sci 63:24–30CrossRefGoogle Scholar
  36. Tasch P (1963) Dead and viable fossil salt bacteria. UnivWichita Bull 39:3–7Google Scholar
  37. Truzzi M (1978) On the extraordinary: an attempt at clarification. Zetetic Scholar 1:11Google Scholar
  38. Vreeland RH, Powers DW (1998) Microbiological considerations for sampling ancient salt formations. In Oren A (ed) Biology and geochemistry of hypersaline environments. CRC Press Series on Life in Extreme and Unusual Environments. Chap. 5, p 53–73Google Scholar
  39. Vreeland RH, Rosenzweig WD (1998) Microorganisms in ancient salt formations: Possibilities and potentials. In: Seckbach J (ed) Enigmatic microorganisms and extreme environments. Kluwer, DordrechtGoogle Scholar
  40. Vreeland RH, Piselli AF Jr, McDonnough S, Meyers SS (1998) Distribution and diversity of halophilic bacteria in a subsurface salt formation. Extremophiles 2:321–331CrossRefGoogle Scholar
  41. Vreeland RH, Rosenzweig WD, Powers DW (2000) Isolation of a 250 million year old bacterium from primary salt crystals. Nature 408:897–900CrossRefGoogle Scholar
  42. Vreeland RH, Rosenzweig WD (2002) The question of uniqueness of ancient bacteria. J Ind Microbiol Biotechnol 28:32–41CrossRefGoogle Scholar
  43. Vreeland RH (2006) Extraction of microorganisms from ancient crystals. In: Rainey F, Oren A (eds) Methods in microbiology, vol. 35. Elsevier, The Netherlands, p 553–567CrossRefGoogle Scholar
  44. Vreeland RH, Lowenstein T, Timofeeff M, Satterfield C, DiFerdinando J, Jones J, Monson A, Rosenzweig WD, Cho BC, Park JS, Wallace A, Grant WD (2007) The isolation of live 125 million year old haloarchaea. Geomicrobiol J 24:274–282CrossRefGoogle Scholar
  45. Zolensky ME, Bodnar RJ, Gibson EK, Nyquist LE, Reese Y, Shih CY, Wiesman H (1999) Asteroidal water within fluid inclusion bearing halite in an H5 chondrite Monahans (1998). Science 285:1377–1379CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  1. 1.Belle HavenUSA

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