Bioassociation of uranium on halophilic archaea analyzed with microscopic and spectroscopic methods

Salt rock is one potential host rock formation for the final disposal of radioactive waste in Germany. Next to geochemical and physical parameters in salt rock indigenous microorganisms have to be taken into account for the safety performance of the deep geological repository. The halophilic archaeon Halobacterium noricense DSM-15987 is indigenous in salt rock and was originally isolated from a salt mine in Austria [1]. This extreme halophilic archaeon was used to study its interactions with uranium. ICP-MS analyses of the supernatant were performed to document the bioassociation kinetic of uranium to cells of Hbt. noricense, which showed a so far unknown behavior. More detailed information about the formed complexes were gained by using different spectroscopic and microscopic methods. All experiments with Hbt. noricense have to be performed at 3 M NaCl which leads to a limited selection of methods or makes their application complicated. Hence, different approaches to generate samples for electron microscopy have been performed and will be the focus of the talk. For example preparation steps like fixation as well as washing steps to get rid of the salt have been varied to obtain SEM and TEM samples which are unaffected by the preparation steps. The used electron microscopic methods have been a useful tool for various reasons. First, by using SEM coupled with EDX a localization of uranium was possible. Second, changes in cell agglomeration as well as uranium bioassociation over time were visible supporting the so far unknown multi-stage bioassociation process. In combination with spectroscopic tools like infrared spectroscopy and laser fluorescence spectroscopy the applied microscopic methods were essential for a better understanding of the bioassociation process of uranium to cells of the halophilic archaeon.