Bacterial diversity in Mont Terri Opalinus Clay and the influence of the bacterial Sporomusa sp. isolate on plutonium speciation


Bacterial diversity in Mont Terri Opalinus Clay and the influence of the bacterial Sporomusa sp. isolate on plutonium speciation

Moll, H.; Lütke, L.; Bachvarova, V.; Geissler, A.; Selenska-Pobell, S.; Bernhard, G.

The concept of geological disposal of nuclear waste comprises a detailed knowledge concerning potential host rock formations. One of such formations is the Opalinus Clay geologic layer of the Mont Terri Underground Rock Laboratory (Switzerland). Dominant bacterial strains from sites destined for future nuclear waste deposition have to be identified and investigated regarding their interaction mechanisms with soluble actinide ions.
For the first time microbial total DNA (tDNA) was isolated from 50 g unperturbed Mont Terri Opalinus Clay. Analysis of the tDNA revealed that the bacterial community of the unperturbed Opalinus Clay is dominated by representatives of Firmicutes, Betaproteobacteria, and Bacteriodetes. Representatives of Firmicutes completely overgrow the other members of the community after treatment of the clay with R2A medium. Bacteria isolated from Mont Terri Opalinus Clay on R2A medium were affiliated with different Sporomusa spp., Paenibacillus spp., and Clostridium spp..
After isolation, characterization, and cultivation, we studied the unknown interaction between plutonium in mixed oxidation states and cell-suspensions of one of the Sporomusa sp. MT-2 isolates. Accumulation experiments were performed in order to obtain information about the amount of Pu bound by the bacteria in dependence on the contact time and the initial plutonium concentration. The 242-Pu present in blank (no cells added), supernatant, and washed biomass suspension at pH 0 was analyzed using UV-vis-NIR spectroscopy, solvent extraction, and liquid scintillation counting (LSC). In addition experiments were performed by adding an electron donor (Na-pyruvate) in two concentrations 0.1 mM and 10 mM.
The amount of Pu sorbed by Sporomusa sp. cells increased with time. Steady state conditions were reached after approximately 200 h. The data could be successfully fitted to a bi-exponential law. The amount of Pu associated with Sporomusa sp. cells depends on the initial 242-Pu concentration. In the first step, a fast binding of the Pu(VI) and Pu(IV)-polymers onto the biomass occurred. Solvent extractions showed that 92 % of the initially present Pu(VI) is reduced to Pu(V) due to the activity of the cells within the first 48 h of contact time (no electron donor added). The corresponding redox potential in the cell suspensions dropped down to 300 mV compared to 780 mV measured in the blanks. Most of the formed Pu(V) dissolves from the cell envelope back to the aqueous solution due to the weak complexing properties of Pu(V). Good binding properties of Pu(IV)-polymers on functional groups of the Sporomusa sp. cell envelope were found (immobilization). In contrast to earlier measurements with Pseudomonas fluorescens (CCUG 32456A) cells clear indications for increased amounts of Pu(IV) and Pu(III) on the Sporomusa biomass were observed. The Pu oxidation state distributions as a function of time will be discussed in detail and the results of the system without addition of electron donors will be compared with the electron donor supplemented systems. Here differences in the Pu interaction mechanism were found.

Keywords: plutonium; bacteria; Sporomusa; speciation; absorption spectroscopy; solvent extraction

  • Contribution to proceedings
    MIGRATION 2013 - 14th International Conference on the Chemistry and Migration Behaviour of Actinides and Fission Products in the Geosphere, 08.-13.09.2013, Brighton, United Kingdom
    Proceedings of MIGRATION 2013
  • Poster
    MIGRATION 2013 - 14th International Conference on the Chemistry and Migration Behaviour of Actinides and Fission Products in the Geosphere, 08.-13.09.2013, Brighton, United Kingdom

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Publ.-Id: 18520