Perchlorate and nitrate were reduced simultaneously in fixed biofilm reactors. Reduction of 1000 mu g L-1 perchlorate decreased slightly with the addition of 10-16 mg L-1 NO3-N when excess acetate was supplied while denitrification was complete. When influent acetate was reduced by 50% to well below the stoichiometric requirement, perchlorate reduction decreased by 70% while denitrification decreased by only 20%, suggesting that competition for electrons by nitrate was a factor in inhibition. Reduction of nitrate was favored over perchlorate, even though reactor biofilm had been enriched under perch lorate-reducing conditions for 10 months. When excess acetate was restored, perchlorate and nitrate returned to initial levels. The average most probable numbers of perchlorate- and nitrate-reducing bacteria during excess substrate operation were not significantly different and ranged between 2.0 x 10(5) and 7.9 x 10(5) cells cm(-2) media surface area. The effect of nitrate on chloride generation by suspensions of perchlorate-reducing populations was studied using a chloride ion probe. The rate of reduction of 2 mM perchlorate decreased by 30% in the presence of 2 mM nitrate when excess acetate was added. When acetate was limited, perchlorate reduction decreased by 70% in the presence of equi-molar nitrate. (C) 2008 Elsevier B.V. All rights reserved.