Microbial processes have shown promise for the remediation of uranium and nitrate in groundwater impacted by uranium mine tailings. This study investigated the inhibitory impact of uranium(VI) towards different microbial populations in anaerobic biofilms, including methanogenic, denitrifying, and uranium-reducing microorganisms, which are commonly found at uranium bioremediation sites. Results of batch activity bioassays indicated a very distinct level of toxicity depending on the targeted microbial community. U(VI) caused severe inhibition of acetoclastic methanogenesis as indicated by a 50 % inhibiting concentration (IC50) of only 0.16 mM. Denitrifying populations were also impacted by uranium, but their sensitivity depended on the electron donor utilized. Sulfur-oxidizing denitrifiers were the least affected (IC50 for denitrification activity = 0.32 mM), followed by H2- and acetate-utilizing denitrifiers (IC50 of 0.20 and 0.15 mM, respectively). In contrast, exposure to U(VI) concentrations up to 1.0 mM did not inhibit the rate of U(VI) bioreduction with H2 as electron donor in the presence or absence of nitrate. On the contrary, a considerable increase in the uranium-reducing activity of the denitrifying and methanogenic mixed cultures was observed with increasing uranium concentrations. The results suggest that microorganisms responsible for U(V) reduction could tolerate much higher uranium concentrations compared to the other microbial populations assayed.