Activated sludges were acclimated to p-nitrophenol (PNP) in two operational modes, a batch and a continuous. The operational mode of the PNP acclimation of activated sludges strongly affected the physiological characteristics of predominant microorganisms responsible for PNP degradation. Predominant PNP degraders in the sludge in batch mode (Sludge B) had lower PNP affinity and were relatively insensitive to PNP concentration. Those of the sludge in continuous mode (Sludge C), on the other hand, had very high PNP affinity and were sensitive to PNP. MPN enumeration of PNP degraders in sludge B and C using media with different PNP concentrations (0.05, 0.2, 0.5 and 2.0 mM) supported the above results. Medium with 0.2 mM of PNP did not recover PNP degraders in sludge C well, while it recovered PNP degraders in sludge B as well as the medium with 0.05 mM did. When switching from one operational mode to the other, the predominant population in sludge B shifted to the sensitive group, but that of sludge C did not shift at the given loading of PNP, showing relative resistance to inhibitive concentration.