Influence of uranium (VI) on the metabolic activity of stable multispecies biofilms studied by oxygen microsensors and fluorescence microscopy

The effect of uranium added in ecologically relevant concentrations (1×10-5 M and 1×10-6 M) to stable multispecies biofilms was studied by electrochemical oxygen microsensors with tip diameters of 10 µm and by confocal laser fluorescence microscopy (CLSM). The microsensor profile measurements in the stable multispecies biofilms exposed to uranium showed that the oxygen concentration decreased faster with increasing biofilm depth compared to the uranium free biofilms. In the uranium containing biofilms, the oxygen consumption, calculated from the steady-state microprofiles, showed high consumption rates of up to 68 nmol cm-3s-1 in the top layer (0 - 70 µm) and much lower consumption rates in the lower zone of the biofilms. Staining experiments with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and 4,6-diamidino-2-phenylindole (DAPI) confirmed the high respiratory activities of the bacteria in the upper layer. Analysis of the amplified 16S rRNA gene fragments showed that the addition of uranium in ecological relevant concentrations did not change the bacterial diversity in the stable multispecies biofilms and is therefore not responsible for the different oxygen profiles in the biofilms. The fast decrease in the oxygen concentrations in the biofilm profiles showed that the bacteria in the top region of the biofilms, i.e. the metabolically most active biofilms zone, battle the toxic effects of aqueous uranium with an increased respiratory activity. This increased respiratory activity results in O2 depleted zones closer to the biofilm air interface which triggers redox processes leading to a precipitation of uranium(IV) solids and consequently to a removal of uranium from the aqueous phase.