Comparison of uranium(VI) removal by Shewanella oneidensis MR-1 in flow and batch reactors

To better understand the interactions among metal contaminants, nutrients, and microorganisms in subsurface fracture-flow systems, biofilms of pure culture of Shewanella oneidensis MR-1 were grown in six fracture-flow reactors (FFRs) of different geometries. The spatial and temporal distribution of uranium and bacteria were examined using a tracer dye (brilliant blue FCF) and microscopy. The results showed that plugging by bacterial cells was dependent on the geometry of the reactor and that biofilms grown in FFRs had a limited U(VI)-reduction capacity. To quantify the U(VI)-reduction capacity of biofilms, batch experiments for U(VI) reduction were performed with repetitive U(VI) additions. U(VI)-reduction rates of stationary phase cultures decreased after each U(VI) addition. After the fourth U(VI) addition, stationary phase cultures treated with U(VI) with and without spent medium yielded gray and black precipitates, respectively. These gray and black U precipitates were analyzed using high-resolution transmission electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray diffraction. Data for randomly selected areas of black precipitates showed that reduced U particles (3-6 nm) were crystalline, whereas gray precipitates were a mixture of crystalline and amorphous solids. Results obtained in this study, including a dramatic limitation of S. oneidensis MR-1 and its biofilms to reduce U(VI) and plugging of FFRs, suggest that alternative organisms should be targeted for stimulation for metal immobilization in subsurface fracture-flow systems.