Acetate Limitation and Nitrite Accumulation during Denitrification

Nitrite accumulated in denitrifying activated sludge mixed liquor when the carbon and electron source, acetate, was limited. If acetate was added to obtain a carbon-to-nitrogen (C:N) ratio in the range of 2:1 to 3:1, nitrate was completely consumed at the same rate with no nitrite accumulation, indicating that nitrate concentration controlled the respiration rate as long as sufficient substrate was present. However, when acetate was reduced to a C:N ratio of 1:1, while nitrate continued to be consumed, >50% of the initial nitrate-nitrogen accumulated as nitrite and 29% persisted as nitrite throughout an endogenous denitrification period of 8–9 h. While nitrite accumulated during acetate-limited denitrification, the specific nitrate reduction rate increased significantly compared with the rate when excess acetate was provided as follows: 0.034 mg-NO3-N/mg-mixed liquid volatile suspended solids/h versus 0.023 mg-NO3-N/mg-mixed liquid volatile suspended solids/h, respectively. This may be explained by nitrate respiration out-competing nitrite respiration for limited acetate electrons. Complete restoration of balanced denitrification and elimination of nitrite accumulation during denitrification required several weeks after the C:N ratio was increased back to 2:1.