This study investigates the response of activated sludge that is fed with proteinaceous feed to silver (Ag + ) and continuous nanosilver (AgNPs) input. Substrate removal (COD) and respiration of activated sludge were examined in two reactors fed with only peptone as an organic substrate and inorganics. Results showed that substrate removal was not affected by the high concentrations of Ag + and AgNP neither in short-nor in long-term. It was concluded that peptone in the feed formed complexes with silver and reduced its negative effects. Also, in long-term, AgNP lost its effectiveness upon continuous aeration. Moreover, in long-term, biomass protected itself from the inhibitory effect of AgNP by regulating the production of Extracellular Polymeric Substances (EPS). Therefore, no negative impact of silver and nanosilver on substrate removal and change in sludge characteristics would be expected.