Environmental Science and Pollution Research

, Volume 26, Issue 9, pp 9352–9364 | Cite as

Association of Eu(III) and Cm(III) onto an extremely halophilic archaeon

  • Miriam Bader
  • Henry Moll
  • Robin Steudtner
  • Henry Lösch
  • Björn Drobot
  • Thorsten Stumpf
  • Andrea CherkoukEmail author
Research Article


In addition to geological, geochemical, and geophysical aspects, also, microbial aspects have to be taken into account when considering the final storage of high-level radioactive waste in a deep geological repository. Rock salt is a potential host rock formation for such a repository. One indigenous microorganism, that is, common in rock salt, is the halophilic archaeon Halobacterium noricense DSM15987T, which was used in our study to investigate its interactions with the trivalent actinide curium and its inactive analogue europium as a function of time and concentration. Time-resolved laser-induced fluorescence spectroscopy was applied to characterize formed species in the micromolar europium concentration range. An extended evaluation of the data with parallel factor analysis revealed the association of Eu(III) to a phosphate compound released by the cells (F2/F1 ratio, 2.50) and a solid phosphate species (F2/F1 ratio, 1.80). The association with an aqueous phosphate species and a solid phosphate species was proven with site-selective TRLFS. Experiments with Cm(III) in the nanomolar concentration range showed a time- and pCH+-dependent species distribution. These species were characterized by red-shifted emission maxima, 600–602 nm, in comparison to the free Cm(III) aqueous ion, 593.8 nm. After 24 h, 40% of the luminescence intensity was measured on the cells corresponding to 0.18 μg Cm(III)/gDBM. Our results demonstrate that Halobacterium noricense DSM15987T interacts with Eu(III) by the formation of phosphate species, whereas for Cm(III), a complexation with carboxylic functional groups was also observed.


Halophilic archaeon Halobacterium noricense DSM15987T Europium and curium bioassociation Luminescence spectroscopy at high salinity Eu/Cm TRLFS Final storage of high-level radioactive waste 



The authors are indebted to the U.S. Department of Energy, Office of Basic Energy Sciences, for the use of 248Cm via the transplutonium element production facilities at Oak Ridge National Laboratory; 248Cm was made available as part of collaboration between HZDR and the Lawrence Berkeley National Laboratory (LBNL).


  1. Andrei AS, Banciu HL, Oren A (2012) Living with salt: metabolic and phylogenetic diversity of archaea inhabiting saline ecosystems. FEMS Microbiol Lett 330:1–9CrossRefGoogle Scholar
  2. Atkins PW (1998) Physical chemistry. Oxford University Press, OxfordGoogle Scholar
  3. Bader M, Müller K, Foerstendorf H, Drobot B, Schmidt M, Musat N, Swanson JS, Reed DT, Stumpf T, Cherkouk A (2017) Multistage bioassociation of uranium onto an extremely halophilic archaeon revealed by a unique combination of spectroscopic and microscopic techniques. J Hazard Mater 327:225–232CrossRefGoogle Scholar
  4. Bader M, Müller K, Foerstendorf H, Schmidt M, Simmons KA, Swanson JS, Reed DT, Stumpf T, Cherkouk A (2018) Comparative analysis of uranium bioassociation with halophilic bacteria and archaea. PloSOne 13:e0190953CrossRefGoogle Scholar
  5. Balda R, Fernandez J, Adam JL, Arriandiaga MA (1996) Time-resolved fluorescence-line narrowing and energy-transfer studies in a Eu3+-doped fluorophosphate glass. Phys Rev B 54:12076–12086CrossRefGoogle Scholar
  6. Binnemans K (2015) Interpretation of europium(III) spectra. Coord Chem Rev 295:1–45CrossRefGoogle Scholar
  7. Drobot B, Steudtner R, Raff J, Geipel G, Brendler V, Tsushima S (2015) Combining luminescence spectroscopy, parallel factor analysis and quantum chemistry to reveal metal speciation - a case study of uranyl(VI) hydrolysis. Chem Sci 6:964–972CrossRefGoogle Scholar
  8. Edelstein NM, Klenze R, Fanghänel T, Hubert S (2006) Optical properties of Cm(III) in crystals and solutions and their application to Cm(III) speciation. Coord Chem Rev 150:948–973CrossRefGoogle Scholar
  9. Ekman P, Jäger O (1993) Quantification of subnanomolar amounts of phosphate bound to seryl and threonyl residues in phosphoproteins using alkaline hydrolysis and malachite green. Anal Biochem 214:138–141CrossRefGoogle Scholar
  10. Fendrihan S, Legat A, Pfaffenhuemer M, Gruber C, Weidler G, Gerbl F, Stan-Lotter H (2006) Extremely halophilic archaea and the issue of long-term microbial survival. Rev Environ Sci Biotechnol 5:203–218CrossRefGoogle Scholar
  11. Francis AJ, Gillow JB, Dodge CJ, Harris R, Beveridge TJ, Papenguth HW (2004) Uranium association with halophilic and non-halophilic bacteria and archaea. Radiochim Acta 92:481–488CrossRefGoogle Scholar
  12. Gadd GM (2009) Biosorption: critical review of scientific rationale, environmental importance and significance for pollution treatment. J Chem Technol Biotechnol 84:13–28CrossRefGoogle Scholar
  13. Gaft M, Reisfeld R, Panczer G, Shoval S, Champagnon B, Boulon G (1997) Eu3+ luminescence in high-symmetry sites of natural apatite. J Lumin 72-4:572–574CrossRefGoogle Scholar
  14. Gillow JB, Dunn M, Francis AJ, Lucero DA, Papenguth HW (2000) The potential of subterranean microbes in facilitating actinde migration at the Grimsel Test Site and Waste Isolation Pilot Plant. Radiochim Acta 88:769–774CrossRefGoogle Scholar
  15. Görller-Walrand C, Fluyt L, Ceulemans A, Carnall W (1991) Magnetic dipole transitions as standards for Judd–Ofelt parametrization in lanthanide spectra. J Chem Phys 95:3099–3106CrossRefGoogle Scholar
  16. Gramain A, Diaz GC, Demergasso C, Lowenstein TK, McGenity TJ (2011) Archaeal diversity along a subterranean salt core from the Salar Grande (Chile). Environ Microbiol 13:2105–2121CrossRefGoogle Scholar
  17. Gruber C, Legat A, Pfaffenhuemer M, Radax C, Weidler G, Busse HJ, Stan-Lotter H (2004) Halobacterium noricense sp. nov., an archaeal isolate from a bore core of an alpine Permian salt deposit, classification of Halobacterium sp. NRC-1 as a strain of H. salinarum and emended description of H. salinarum. Extremophiles 8:431–439CrossRefGoogle Scholar
  18. Heller A, Barkleit A, Foerstendorf H, Tsushima S, Heim K, Bernhard G (2012) Curium (III) citrate speciation in biological systems: a europium (III) assisted spectroscopic and quantum chemical study. Dalton Trans 41:13969–13983CrossRefGoogle Scholar
  19. Holliday K, Handley-Sidhu S, Dardenne K, Renshaw J, Macaskie L, Walther C, Stumpf T (2012) A new incorporation mechanism for trivalent actinides into bioapatite: a TRLFS and EXAFS study. Langmuir 28:3845–3851CrossRefGoogle Scholar
  20. Horrocks WD Jr, Sudnick DR (1979) Lanthanide ion probes of structure in biology. Laser-induced luminescence decay constants provide a direct measure of the number of metal-coordinated water molecules. J Am Chem Soc 101:334–340CrossRefGoogle Scholar
  21. Hunsche U, Hampel A (1999) Rock salt - the mechanical properties of the host rock material for a radioactive waste repository. Eng Geol 52:271–291CrossRefGoogle Scholar
  22. Jordan N, Demnitz M, Losch H, Starke S, Brendler V, Huittinen N (2018) Complexation of trivalent lanthanides (Eu) and actinides (Cm) with aqueous phosphates at elevated temperatures. Inorg Chem 57:7015–7024CrossRefGoogle Scholar
  23. Karbowiak M, Hubert S (2000) Site-selective emission spectra of Eu3+: Ca-5(PO4)(3)F. J Alloys Compd 302:87–93CrossRefGoogle Scholar
  24. Kim JI, Rhee DS, Wimmer H, Buckau G, Klenze R (1993) Complexation of trivalente actinide ions (Am3+, Cm3+) with humic-acid - a comparison of different experimental methods. Radiochim Acta 62:35–43CrossRefGoogle Scholar
  25. Kimura T, Choppin GR (1994) Luminescence study on determination of the hydration number of Cm (III). J Alloys Compd 213:313–317CrossRefGoogle Scholar
  26. Kimura T, Choppin GR, Kato Y, Yoshida Z (1996) Determination of the hydration number of Cm(III) in various aqueous solutions. Radiochim Acta 72:61–64CrossRefGoogle Scholar
  27. Kimura T, Kato Y (1998) Luminescence study on hydration states of lanthanide (III)–polyaminopolycarboxylate complexes in aqueous solution. J Alloys Compd 275:806–810CrossRefGoogle Scholar
  28. Klimmek S (2003) Charakterisierung der Biosorption von Schwermetallen an Algen. Technischen Universität, BerlinGoogle Scholar
  29. Kuke S, Marmodee B, Eidner S, Schilde U, Kumke MU (2010) Intramolecular deactivation processes in complexes of salicylic acid or glycolic acid with Eu(III). Spectroc Acta Pt A-Molec Biomolec Spectr 75:1333–1340CrossRefGoogle Scholar
  30. Kümmel R, Worch E (1990) Adsorption aus wässrigen Lösungen. Dt. Verl. für Grundstoffindustrie, LeipzigGoogle Scholar
  31. Leuko S, Legat A, Fendrihan S, Stan-Lotter H (2004) Evaluation of the LIVE/DEAD BacLight kit for detection of extremophilic archaea and visualization of microorganisms in environmental hypersaline samples. Appl Environ Microbiol 70:6884–6886CrossRefGoogle Scholar
  32. Lloyd JR, Macaskie LE (2002): Biochemical basis of microbe-radionuclide interactions. In: Keith-Roach MJ , Livens FR (Editors), Interactions of microorganisms with radionuclides. Radioactivity in the environment. Elsevier, pp. 313–342Google Scholar
  33. Lumetta GJ, Thompson MC, Pennemann RA, Eller PG (2008 ): Curium. In: Morss LR, Edelstein NM, Fuger J (Editors), The chemistry of the actinide and transactinide chemistry - Third Edition. Springer, pp. 1397–1443Google Scholar
  34. Lütke L 2013: Interaction of selected actinides (U, Cm) with bacteria relevant to nuclear waste disposal., TU DresdenGoogle Scholar
  35. Macaskie LE, Bonthrone KM, Yong P, Goddard DT (2000) Enzymically mediated bioprecipitation of uranium by a Citrobacter sp.: a concerted role for exocellular lipopolysaccharide and associated phosphatase in biomineral formation. Microbiology-Sgm 146:1855–1867CrossRefGoogle Scholar
  36. McGenity TJ, Gemmell RT, Grant WD, Stan-Lotter H (2000) Origins of halophilic microorganisms in ancient salt deposits. Environ Microbiol 2:243–250CrossRefGoogle Scholar
  37. Moll H, Stumpf T, Merroun M, Rossberg A, Selenska-Pobell S, Bernhard G (2004) Time-resolved laser fluorescence spectroscopy study on the interaction of curium(III) with Desulfovibrio aspoensis DSM 10631. Environ Sci Technol 38:1455–1459CrossRefGoogle Scholar
  38. Moll H, Geipel G, Bernhard G (2005) Complexation of curium(III) by adenosine 5′-triphosphate (ATP): a time-resolved laser-induced fluorescence spectroscopy (TRLFS) study. Inorg Chim Acta 358:2275–2282CrossRefGoogle Scholar
  39. Moll H, Glorius M, Barkleit A, Rossberg A, Bernhard G (2009) The mobilization of actinides by microbial ligands taking into consideration the final storage of nuclear waste: interactions of selected actinides U(VI), Cm(III), and Np(V) with pyoverdins secreted by Pseudomonas fluorescens and related model compounds. In: Forschungszentrum Dresden-Rossendorf. Dresden, GermanyGoogle Scholar
  40. Moll H, Brendler V, Bernhard G (2011) Aqueous curium(III) phosphate species characterized by time-resolved laser-induced fluorescence spectroscopy. Radiochim Acta 99:775–782CrossRefGoogle Scholar
  41. Moll H, Lutke L, Barkleit A, Bernhard G (2013) Curium(III) speciation studies with cells of a groundwater strain of Pseudomonas fluorescens. Geomicrobiol J 30:337–346CrossRefGoogle Scholar
  42. Moll H, Lütke L, Bachvarova V, Cherkouk A, Selenska-Pobell S, Bernhard G (2014) Interactions of the Mont Terri Opalinus Clay isolate Sporomusa sp. MT-2.99 with curium (III) and europium (III). Geomicrobiol J 31:682–696CrossRefGoogle Scholar
  43. Ozaki T, Gillow JB, Francis AJ, Kimura T, Ohnuki T, Yoshida Z (2002) Association of Eu(III) and Cm(III) with Bacillus subtilis and Halobacterium salinarum. J Nucl Sci Techn Suppl 3:950–953CrossRefGoogle Scholar
  44. Ozaki T, Gillow JB, Kimura T, Ohnuki T, Yoshida Z, Francis AJ (2004) Sorption behavior of europium(III) and curium(III) on the cell surfaces of microorganisms. Radiochim Acta 92:741–748CrossRefGoogle Scholar
  45. Piriou B, Fahmi D, Dexpertghys J, Taitai A, Lacout JL (1987) Unusual fluorescent properties of Eu3+ in oxyapatites. J Lumin 39:97–103CrossRefGoogle Scholar
  46. Plancque G, Moulin V, Toulhoat P, Moulin C (2003) Europium speciation by time-resolved laser-induced fluorescence. Anal Chim Acta 478:11–22CrossRefGoogle Scholar
  47. Selenska-Pobell S, Merroun ML (2010) Accumulation of heavy metals by microorganisms: bimineralization and nanocluster formation. In: König H, Claus H, Varma A (eds) Prokaryotic cell wall compounds - structure and biochemistry. Springer-Verlag, Berlin Heidelberg, pp 483–500CrossRefGoogle Scholar
  48. Swanson JS, Reed DT, Ams DA, Norden D, Simmons KA (2012): Status report on the microbial characterization of halite and groundwater samples from the WIPP. Status Report LA-UR-12-22824Google Scholar
  49. 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
  50. Yong P, Macaskie LE, Sammons RL, Marquis PM (2004) Synthesis of nanophase hydroxyapatite by a Serratia sp from waste-water containing inorganic phosphate. Biotechnol Lett 26:1723–1730CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Resource EcologyHelmholtz-Zentrum Dresden - RossendorfDresdenGermany
  2. 2.Max Planck Institute of Molecular Cell Biology and Genetics, Tang LabDresdenGermany

Personalised recommendations