Determination of structural components, e.g. exopolymeric substances (EPS) within biofilms and their effect on the immobilisation of actinides (U) - Determination of pH, Eh, and dissolved oxygen and their influence on the actinide speciation

Fluorescent uranium(V) and uranium(VI) particles were observed for the first time in vivo by a combined laser fluorescence spectroscopy and confocal laser scanning microscopy approach in a living multispecies biofilm grown on biotite plates. Anaysis of amplified 16S rRNA fragments and fluorescence in situ hybridization were used to characterize the biofilm communities. Laser fluorescence spectroscopy was used to identify these particles. The particles showed either a characteristic fluorescence spectrum in the wavelength range of 415-475 nm, indicative for uranium(V), or in the range of 480-560 nm, which is typical for uranium(VI). Particles of uranium(V) as well as uranium(VI) were simultaneously observed in the biofilms. These uranium particles were attributed for uranium(VI) to biologically mediated precipitation and for uranium(V) to redox processes taking place within the biofilms. Concentration profiles of oxygen versus biofilm depths were measured in the biofilms by electrochemical microsensors with a tip diameter of 10µm. A motor-driven micromanipulator was used for moving downwards through the biofilm in 20 or 50µm steps. The microsensor results clearly showed that the vertical profiles of the O2 concentration within the biofilms are affected by the presence as well as by the concentration of U(VI) in the culture media. In the absence of uranium the O2 concentration in the well aerated biofilm decreased slightly. In contrast, O2 concentrations in the biofilms, which were exposed to different concentrations of uranium, decreased with increasing uranium concentration.