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Anaerobic bioreactors that can support simultaneous microbial processes of denitrification and methanogenesis are of interest to nutrient nitrogen removal. However, an important concern is the potential toxicity of nitrate (NO(3) (-)) and nitrite (NO(2) (-)) to methanogenesis. The methanogenic toxicity of the NO (x) (-) compounds to anaerobic granular biofilms and municipal anaerobic digested sludge with two types of substrates, acetate and hydrogen, was studied. The inhibition was the severest when the NO (x) (-) compounds were still present in the media (exposure period). During this period, 95% or greater inhibition of methanogenesis was evident at the lowest concentrations of added NO(2) (-) tested (7.6-10.2 mg NO(2) (-)-N l(-1)) or 8.3-121 mg NO(3) (-)-N l(-1) of added NO(3) (-), depending on substrate and inoculum source. The inhibition imparted by NO(3) (-) was not due directly to NO(3) (-) itself, but instead due to reduced intermediates (e.g., NO(2) (-)) formed during the denitrification process. The toxicity of NO (x) (-) was found to be reversible after the exposure period. The recovery of activity was nearly complete at low added NO (x) (-) concentrations; whereas the recovery was only partial at high added NO (x) (-) concentrations. The recovery is attributed to the metabolism of the NO (x) (-) compounds. The assay substrate had a large impact on the rate of NO(2) (-) metabolism. Hydrogen reduced NO(2) (-) slowly such that NO(2) (-) accumulated more and as a result, the toxicity was greater compared to acetate as a substrate. The final methane yield was inversely proportional to the amount of NO (x) (-) compounds added indicating that they were the preferred electron acceptors compared to methanogenesis.

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