[en] For the first time microbial tDNA could be isolated from 50 g unperturbed Mont Terri Opalinus Clay. Based on the analysis of the tDNA the bacterial diversity of the unperturbed clay is dominated by representatives of Firmicutes, Betaproteobacteria, and Bacteriodetes. Firmicutes also dominate after treatment of the clay with R2A medium. Bacteria isolated from Mont Terri Opalinus Clay on R2A medium were related to Sporomusa spp., Paenibacillus spp., and Clostridium spp. All further investigations are concentrated on the unique isolates Sporomusa sp. MT-2 and Paenibacillus sp. MT-2. Cells of the type Sporomusa sp. MT-2 and Paenibacillus sp. MT-2 were comprehensively analyzed in terms of growing, morphology, functional groups of the cell envelope, and cell membrane structure. Strong actinide(An)/lanthanide(Ln)-interactions with the Opalinus Clay isolates and the Aespoe-strain Pseudomonas fluorescens (CCUG 32456) could be determined within a broad pH range (2-8). The metals bind as a function of pH on protonated phosphoryl, carboxyl and deprotonated phosphoryl sites of the respective cell membrane. The thermodynamic surface complexation constants of bacterial An/Ln-species were determined and can be used in modeling programs. Depending on the used An different interaction mechanisms were found (U(VI): biosorption, partly biomineralisation; Cm(III): biosorption, indications for embedded Cm(III); Pu: biosorption, bioreduction and indications for embedded Pu). Different strategies of coping with U(VI) were observed comparing P. fluorescens planktonic cells and biofilms under the chosen experimental conditions. An enhanced capability of the biofilm to form meta-autunite in comparison to the planktonic cells was proven. Conclusively, the P. fluorescens biofilm is more efficient in U(VI) detoxification. In conclusion, Mont Terri Opalinus Clay contains bacterial communities, that may influence the speciation and hence the migration behavior of selected An/Ln under environmental conditions.

[en] The yeast KS5 (Rhodosporidium toruloides) was isolated by culture dependent method directly from the flooding water in Koenigstein (Germany). To compare the U tolerance and immobilization ability of the isolate a reference culture DSM 10134 (R. toruloides) was applied. Both cultures displayed the ability to tolerate high amounts of U, in contrast the reference KS5 showed a six-fold higher U tolerance in comparison to the reference strain. U immobilization studies displayed that both organisms are able to remove high amounts. The flooding water in Koenigstein has to be cleaned up for many years by an intensive waste water treatment plant. Possibly, with the help of natural occuring microorganisms the flooding water could be cleaned up using in situ bioremediation.

[en] In EF-TEM/EELS studies it was shown that U(VI) is sorbed mainly on the outer membrane of Acidovorax facilis. The results are supported by TRLFS measurements, which were performed on the pellet of the cells. In comparison to reference spectra of some cell membrane components, the measured emission spectra of the A. facilis pellet show the best agreement with those of the Uranyl-lipopolysaccharide-complex. Hence, it can be concluded that phosphoryl groups may be the main binding sites for uranyl, located in the lipopolysaccharide unit in the outer membrane.

[en] The former uranium mine Koenigstein (Germany) is currently in the process of controlled flooding by reason of remediation purposes. However, the flooding water still contains high concentrations of uranium and other heavy metals. For that reason the water has to be cleaned up by a conventional waste water treatment plant. The aim of this study was to investigate the interactions between anaerobic microorganisms and uranium for possible bioremediation approaches, which could be an great alternative for the intensive and expensive waste water treatment. EXAFS (extended X-ray absorption fine structure) and XANES (X-ray absorption near edge structure) measurements were performed and revealed a complete reduction of U(VI) to U(IV) only by adding 10 mM glycerol.

[en] XAS spectra of U and Np sorption biogenic ferrihydrite samples were compared to abiotic samples. The k"3-weighted χ-spectrum and its Fourier-transform of the studied biogenic ferrihydrite sample bears close resemblance to the bidentate edge-sharing innersphere sorption "1E complex, which is the main sorption species on abiotic ferrihydrite. Based on the shell fit analysis, the distances of the coordination shells U-O_e_q, U-O_a_x, and U-Fe are similar to those determined for abiotic ferrihydrite samples.

[en] In crystalline rock, the dominant transport medium for radionuclides is groundwater flowing through subsurface fractures. Since groundwater is containing microorganisms, fracture surfaces support biological growth of microbial communities, the so-called bio-films. The microbial diversity of these bio-films depends on the microbial consortia and the chemical composition of the fracture water. Subsurface bio-films have a significant effect on the adsorption capacity of host rock formations by forming a barrier between the rock surface and the groundwater. They can significantly affect subsurface biogeochemical interactions, leading to the immobilization and adsorption of radionuclides. Microbial studies were performed to evaluate the relevance of microbial processes for the immobilization of radionuclides in a deep crystalline repository for high-level radioactive waste. Studies were performed in Olkiluoto, in the rock characterization facility ONKALO in Finland, and in the Aespoe Hard Rock Laboratory (HRL) in Sweden. Massive 5-10-mm thick bio-films were observed in both sites attached to tunnel walls where groundwater was seeping from bedrock fractures. In experiments the effect of uranium on bio-films was studied on site in the ONKALO tunnel by adding UO₂(ClO₄)₂ with a final U-concentration of 1.0x10^-5 M to the fracture water in a self-constructed flow cell by using detached bio-film samples. bio-film specimens collected for transmission electron microscopy studies indicated that uranium in the bio-film was immobilized intracellularly in microorganisms as needle-shaped uranyl phosphate minerals, similar to meta-Autunite (Ca[UO₂]₂[PO₄]₂.10-12H₂O). In contrast, thermodynamic calculation of the theoretical predominant fields of uranium species and time-resolved laser fluorescence spectroscopy showed that the formation of aqueous uranium carbonate species Ca₂UO₂(CO₃)₃ and Mg₂UO₂(CO₃)₃ was predicted due to the high concentration of carbonate in the groundwater. At the Aespoe HRL (Sweden) Gallionella ferruginea dominated bio-films associated with bacteriogenic iron oxides (BIOS) and groundwater were sampled from an in situ continuous flow cell, which has been installed in a cavity of the main access tunnel. In laboratory sorption experiments UO₂(ClO₄)₂ and NpO₂ClO₄ were added to the BIOS bio-films in groundwater under aerobic conditions adjusting a final U(VI) concentration of 1.9x10^-5 M.U(VI) and 3.27x10^-5 M Np(V). The results of the experiments showed that in the BIOS bio-film the ferrous iron-oxidizing and stalk-forming bacterium Gallionella ferruginea is dominating the sorption process. The stalk represents an organic surface upon which Fe oxy-hydroxides can precipitate. Under the given pH conditions the uptake of uranium (85%) and Np (95%) depends predominantly on the high amount of ferrihydrite precipitated onto the stalks. The results showed that the combination of the biological material and Fe oxides created an abundant surface area for bioaccumulation and adsorption of radionuclides. (authors)