Polar Biology

, Volume 37, Issue 1, pp 47–59

Penetration of fluorescent microspheres into the NEEM (North Eemian) Greenland ice core to assess the probability of microbial contamination

Original Paper

Abstract

The validation of microbiological results from non-aseptically drilled deep ice cores is challenging because exogenous microbial cells can be transported into the core interior and compromise the existing microbial populations. The NEEM (North Eemian) ice core in Greenland provided a first-time opportunity to use fluorescent microspheres as tracers for assessing potential microbial contamination of glacial ice. We developed specific procedures to coat the surfaces of selected NEEM core samples representing bubbly (93–100 m), brittle (633–644 m) and clathrated (1,730 and 2,050 m) ice with melamine-based carboxylated fluorescent microspheres and tracked periodically their penetration into the core interior for 2.5 years using flow cytometry. Sufficient ice surface coating was achieved by immersing retrieved cores in plastic bags containing suspensions of pre-counted 1- and 10-μm microspheres or by down-hole microsphere deployment in plastic sleeves attached to the drill barrel and liberated during drilling. We examined the relationship between microspheres penetration and ice core depth, structure and time after coating. One consistent observation for all cores (except the brittle ice) was that removing a few millimeters of the outer layer drastically reduced microsphere counts, independent of timing, indicating that penetration was mostly limited to the surface layers. Any deeper penetration was found to be microsphere size dependent. The brittle ice showed significant microsphere penetration possibly due to microfractures. Overall, the use of fluorescent microspheres as tracers and microbial surrogates proved to be a sensitive approach for testing potential contamination during deep core projects.

Keywords

Ice core NEEM Greenland Microbial contamination Fluorescent microspheres Penetration 

References

  1. Alekhina IA, Marie D, Petit J, Lukin V, Zubkov V, Bulat S (2007) Molecular analysis of bacterial diversity in kerosene-based drilling fluid from the deep ice borehole at Vostok. East Antarct FEMS Microbiol Ecol 59:289–299CrossRefGoogle Scholar
  2. Amato P, Doyle S, Christner BC (2009) Macromolecular synthesis by yeasts under frozen conditions. Environ Microbiol 11:589–596PubMedCrossRefGoogle Scholar
  3. Barletta RE, Priscu JC, Mader HM, Jones WL, Roe CH (2012) Chemical analysis of ice vein microenvironments: II. Analysis of glacial samples from Greenland and Antarctica. J Glaciol 58:212, 1109–1118Google Scholar
  4. Binder T, Weikusat I, Freitag J, Garbe CS, Wagenbach D, Kipfstuhl S (2013) Microstructure through an ice sheet. Mater Sci Forum 753:481–484CrossRefGoogle Scholar
  5. Christner B, Mikucki J, Foreman C, Denson J, Priscu J (2005) Glacial ice cores: a model system for developing extraterrestrial decontamination protocols. Icarus 174:284–572CrossRefGoogle Scholar
  6. Colwell ES, Stormberg GJ, Phelbs TJ, Birnbaum SA, McKinley J, Rawson SA, Veverla C, Goodwin S, Long PE, Russell BF, Garland T, Thompson D, Skinner P, Grover S (1992) Innovative techniques for collection of saturated and unsaturated subsurface basalts and sediments for microbiological characterization. J Microbiol Methods 15:279–292CrossRefGoogle Scholar
  7. Dahl-Jensen D, Albert MR, Aldahan A, NEEM Community et al (2013) Eemian interglacial reconstructed from a Greenland folded ice core. Nature 493(7433):489–494CrossRefGoogle Scholar
  8. Dani KGS, Mader HM, Wolff EW, Wadham JL (2012) Modeling the liquid-water veins system within polar ice sheets as a potential microbial habitat. Earth Planet Sci Lett 333–334:238–249CrossRefGoogle Scholar
  9. Gerasimoff M (2003) Drilling fluid observations and recommendations for U.S. Polar Program, Wais cores Drilling Project. Report, Space Science and Engineering Center, University of Wisconsin, Madison, pp 1–33Google Scholar
  10. Gow AJ, Meese DM, Alley RB, Fitzpatrick JJ, Anandadrishnan S, Woods GA, Elder BC (1997) Physical and structural properties of the Greenland Ice Sheet Project 2 ice core: a review. J Geophys Res 102(C12): 26559–26575Google Scholar
  11. Haldeman DL, Amy PS, Russel CE, Jacobson R (1995) Comparison of drilling and mining as methods for obtaining microbiological samples from the deep subsurface. J Microbiol Methods 21:305–316CrossRefGoogle Scholar
  12. Harvey R, Metge D, Sheets R, Jaspers J (2011) Fluorescent microspheres as surrogates in evaluating the efficacy of riverbank filtration for removing Cryptosporidium parvum oocysts and other pathogens. In: Ray C, Shamrukh M (eds) Riverbank filtration for water security in desert countries. Springer, Berlin, pp 81–96Google Scholar
  13. House CH, Cragg BA, Teske A, Leg 210 Scientific Party (2003) Drilling contamination tests during ODP Leg 201 using chemical and particulate tracers. In: D’Hondt SL, Jorgensen BB, Miller DJ et al (eds) Proc ODP, Init Repts, vol 201, pp 1–19Google Scholar
  14. Juck D, Whissell G, Steven B, Pollard W, McKay CP, Greer C, Whyte L (2005) Utilization of fluorescent microspheres and a green fluorescent protein-marked strain for assessment of microbiological contamination of permafrost and ground ice core samples from the Canadian high Arctic. Appl Environ Microbiol 71:1035PubMedCentralPubMedCrossRefGoogle Scholar
  15. Larsen LBC, Sheldon SG, Steffensen JP (2011) NEEM Field season 2011 of deep ice core drilling and core and processing, July 2010. Copenhagen, University of Copenhagen, pp 1–107. http://neem.dk/documentation/2011
  16. Mader HM, Pettitt ME, Wadham JL, Wolff EW, Parkes RJ (2006) Subsurface ice as a microbial habitat. Geology 34:169–172CrossRefGoogle Scholar
  17. Metge DW, Harvey RW, Anders R, Rosenberry DO, Seymour D, Jasperse J (2007) Use of carboxylated microspheres to assess transport potential of Cryptosporidium parvum oocysts at the Russian River water supply facility, Sonoma County, California. Geomicrobiol J 24(3):231–245Google Scholar
  18. Miteva VI, Brenchley JB (2005) Detection and isolation of ultrasmall microorganisms from a 120,000 years old Greenland glacier ice core. Appl Environ Microbiol 71:7806–7818PubMedCentralPubMedCrossRefGoogle Scholar
  19. Phelbs TJ, Frederickson JK (2002) Drilling, coring and sampling subsurface environments. In Hurst et al. (eds) Manual of environmental microbiology, 2nd edn. ASM Press, Washington DC, pp 679–695Google Scholar
  20. Price PB (2000) A habitat for psychrophiles in deep Antarctic ice. Proc Natl Acad Sci USA 97:1247–1251PubMedCrossRefGoogle Scholar
  21. Rasmussen SO, Abbott P, Blunier T, Bourne A, Brook E, Buchardt SL, Buizert C, Chappellaz J, Clausen HB, Cook E, Davies S, Guillevic M, Kipfstuhl S, Laepple T, Dahl-Jensen D, Seierstad IK, Severinghaus JP, Steffensen JP, Stowasser C, Svensson A, Vallelonga P, Vinther BM, Wilhelms F, Winstrup M (2013) A first chronology for the NEEM ice core. Clim Past Discuss 9:2967–3013. doi:10.5194/cpd-9-2967-2013 Google Scholar
  22. Rogers SO, Theraisnathan V, Ma LJ, Zhao Y, Zhang G, Shin S, Castelo J, Starmer W (2004) Comparisons of protocols for decontamination of environmental ice samples for biological and molecular examinations. Appl Environ Microbiol 70:2540–2544PubMedCentralPubMedCrossRefGoogle Scholar
  23. Rogers SO, Ma J, Zhao Y, Theraisnathan V, Shin S G, Zhang G, Catranis CM, Starmer WT, Castelo JD (2005) Recommendations for elimination of contaminants and authentication of isolates in ancient ice cores. In: Castello JD, Rogers SO (eds) Life in ancient ice. Princeton University Press, Princeton, pp 5–22Google Scholar
  24. Rohde RA, Price PB (2007) Diffusion-controlled metabolism for long-term survival of single isolated microorganisms trapped within ice crystals. Proc Natl Acad Sci USA 104(42):16592–16597PubMedCrossRefGoogle Scholar
  25. Smith DC, Spivak AJ, Fisk MR, Haveman SA, Staudigel H (2000) Tracer-based estimates of drilling-induced microbial contamination of deep sea crust. Geomicrobiol J 17:207–219CrossRefGoogle Scholar
  26. Talalay PG (2011) Drilling fluids for deep coring in central Antarctica. Technical report PRC 12-01 Polar Res. Center at Jilin University, China, pp 1–31Google Scholar
  27. Willerslev E, Hansen AJ, Pointar HN (2004) Isolation of nucleic acids and cultures from fossil ice and permafrost. Trends Ecol Evol 9:141–147CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular BiologyThe Pennsylvania State UniversityUniversity ParkUSA
  2. 2.Department of Geosciences, Earth and Environment Systems InstituteThe Pennsylvania State UniversityUniversity ParkUSA

Personalised recommendations