Letter | Published:

Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal

Nature volume 407, pages 897900 (19 October 2000) | Download Citation

Subjects

Abstract

Bacteria have been found associated with a variety of ancient samples1, however few studies are generally accepted due to questions about sample quality and contamination. When Cano and Borucki2 isolated a strain of Bacillus sphaericus from an extinct bee trapped in 25–30 million-year-old amber, careful sample selection and stringent sterilization techniques were the keys to acceptance. Here we report the isolation and growth of a previously unrecognized spore-forming bacterium (Bacillus species, designated 2-9-3) from a brine inclusion within a 250 million-year-old salt crystal from the Permian Salado Formation. Complete gene sequences of the 16S ribosomal DNA show that the organism is part of the lineage of Bacillus marismortui and Virgibacillus pantothenticus. Delicate crystal structures and sedimentary features indicate the salt has not recrystallized since formation. Samples were rejected if brine inclusions showed physical signs of possible contamination. Surfaces of salt crystal samples were sterilized with strong alkali and acid before extracting brines from inclusions. Sterilization procedures reduce the probability of contamination to less than 1 in 10 9.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , & Halobacteria: the evidence for longevity. Extremophiles 2, 279–287 (1998).

  2. 2.

    & Revival and identification of bacterial spores in 25 to 40 million year old Dominican amber. Science 268, 1060–1064 (1995).

  3. 3.

    et al. Halococcus salifodinae sp. nov., an archaeal isolate from an Austrian salt mine. Int. J. Syst. Bacteriol. 44, 774–780 (1994).

  4. 4.

    & in General and Applied Aspects of Halophilic Bacteria. Vol. 201 (ed. Rodriguez-Valera, F.) 53–62 (Plenum, New York, 1991).

  5. 5.

    , & Archaeal halophiles (halobacteria) from two British salt mines. J. Gen. Microbiol. 139, 1077 –1081 (1993).

  6. 6.

    et al. Diversity of microorganisms isolated from amber. Microb. Ecol. 38, 58–68 (1999).

  7. 7.

    et al. Staphylococcus succinus sp. nov., isolated from Dominican amber. Int. J. Syst. Bacteriol. 48, 511– 518 (1998).

  8. 8.

    & in Microbiolology and Biogeochemistry of Hypersaline Environments (ed. Oren, A.) 53–74 (CRC, Boca Raton, Florida, 1999 ).

  9. 9.

    & in Enigmatic and Extreme Microorganisms. (ed. Seckbach, J.) 387– 398 (Kluwer, Delft, 1998).

  10. 10.

    Upper Permian conodonts and other microfossils from the Pinery and Lamar Limestone Members of the Bell Canyon Formation and from the Rustler Formation, west Texas. Thesis, Ohio State Univ. (1978).

  11. 11.

    Paleontology of the Rustler Formation, Culberson County, Texas. J. of Paleontol. 27, 679–702 (1953).

  12. 12.

    , , , & 40/39 Ar and U/Pb SHRIMP dating of latest Permian tephras in the Midland Basin Texas. EOS 77, 794 (1996).

  13. 13.

    , & 40Ar/39Ar dating of langbeinite [K2Mg2(SO4)3] in late Permian evaporites of the Salado Formation, Southeastern New Mexico, USA. Mineral. Mag. 62A, 1253– 1254 (1998).

  14. 14.

    , & The problem of distinguishing between primary and secondary features in evaporites. Sixth Int. Symp. On Salt 1, 11–39 (1983).

  15. 15.

    The fluids in salt. Amer. Mineral. 69, 413 –439 (1984).

  16. 16.

    & Criteria for recognition of salt-pan evaporites. Sedimentol. 32, 627–644 (1985).

  17. 17.

    Origin of depositional cycles in a Permian “saline giant”: the Salado (McNutt zone) evaporites of New Mexico and Texas. Geol. Soc. Am. Bull. 100, 592–608 (1988).

  18. 18.

    & Geological mapping of the air intake shaft at the Waste Isolation Pilot Plant. Report no. DOE/WIPP 90-051, 1–90 (U. S. Department of Energy, Carlsbad NM, 1990).

  19. 19.

    & Syndepositional origin of potash evaporites: petrographic and fluid inclusion evidence. Am. J. Science 290, 1–42 (1990).

  20. 20.

    & in Geological and Hydrological Studies of Evaporites in the Northern Delaware Basin for the Waste Isolation Pilot Plant (WIPP), New Mexico (eds Powers, D. W., Holt, R. M., Beauheim, R. L. & Rempe, N.). Geol. Soc. Am. Guidebook 14, 45–78 (1990).

  21. 21.

    & Synsedimentary dissolution pits of halite of the Permian Salado Formation, southeastern New Mexico. J. Sed. Petrol. 55, 769–773 (1985).

  22. 22.

    , , , & 40Ar/39Ar laser microprobe dating of polyhalite from bedded, late Permian evaporites. EOS 76, S285 (1995).

  23. 23.

    , , & Distribution and diversity of halophilic bacteria in a subsurface salt formation. Extremophiles 2, 321–331 (1998).

  24. 24.

    , , & Development of a protocol to retrieve microorganisms from ancient salt crystals. Geomicrobiol. (in the press).

  25. 25.

    , & E. Cell wall and phospholipid composition and their contribution to the salt tolerance of Halomonas elongata. J. Bacteriol. 160, 879–883 (1984).

  26. 26.

    , , & Bacillus marismortui sp. nov., a new moderately halophilic species from the Dead Sea. Int. J. Syst. Bacteriol. 49, 521–530 ( 1999).

  27. 27.

    et al. Virgibacillus: a new genus to accommodate Bacillus pantothenticus (Proom and Knight 1950). Emended description of Virgibacillus pantothenticus. Int. J. Syst. Bacteriol. 48, 99–106 (1998).

  28. 28.

    , , , & Bacillus salexigens sp. nov., a new moderately halophilic Bacillus species. Int. J. Syst. Bacteriol. 47, 735–741 ( 1997).

  29. 29.

    , , & Gracilibacillus gen nov., with description of Gracilibacillus halotolerans gen. nov. sp. nov.; transfer of Bacillus dipsosauri to Gracilibacillus dipsosauri comb. nov., and Bacillus salexigens to the genus Salibacillus gen. nov., as Salibacillus salexigens comb. nov. Int. J. Syst. Bacteriol. 49, 821–831 (1999).

  30. 30.

    & Analysis of Frankia evolutionary radiation using glnII sequences. FEMS Microbiol. Lett. 177, 29–34 (1999).

Download references

Acknowledgements

The authors acknowledge the following people who helped obtain the crystal samples for this research: D. Belski, N. Rempe, R. Carrasco, T. Garcia, D. Acevedo, S. Britain, E. Keyser, B. Kinsall, A. Morin and T. Padilla. This research was supported by the US National Science Foundation: Life in Extreme Environments Program (EAR Lexen).

Author information

Affiliations

  1. *Department of Biology, West Chester University, West Chester, Pennsylvania 19383 , USA

    • Russell H. Vreeland
    •  & William D. Rosenzweig
  2. †Consulting Geologist, Box 87, Anthony, Texas 79821, USA

    • Dennis W. Powers

Authors

  1. Search for Russell H. Vreeland in:

  2. Search for William D. Rosenzweig in:

  3. Search for Dennis W. Powers in:

Corresponding author

Correspondence to Russell H. Vreeland.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/35038060

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.