Ammonium and nitrite are two substrates of anammox bacteria, but they are also inhibitors under high concentrations. The performance of two anaerobic ammonium-oxidizing (anammox) upflow biofilm (UBF) reactors was investigated. The results show that anammox UBFs become unstable under nitrogen loading rate (NLR) applied higher than 1.0 g/(L.d). The consumptions of acidity in the anammox reaction lead to the increase of pH, which is as high as 8.70-9.05. Free nitrous acid concentration is accompanied to be lower than the affinity constant of anammox bacteria, and then starvation effect appears. Moreover, free ammonia concentration increases to 57-178 mg/L, resulting in inhibitory effect on the anammox bacteria. Both negative effects contribute to the instability of the anammox bioreactors.