We investigated biofouling in ultrafiltration (UF) for drinking water treatment and its control by backwashing with chlorinated-water or pure water. By using sodium azide to suppress biological growth, the relative contribution of biofouling to total fouling was estimated, and its value (5.3-56.0%) varied with the feed water, and increased with the increases of filtration time and membrane flux. The biofouling layer could partially remove biodegradable organic matter and ammonia (32.9-74.2%). Backwashing using chlorinated-water partly inactivated the microorganisms (23.8%) but increased the content of extracellular polymeric substances (7.7%) in the biofouling layer. In contrast, backwashing using pure water led to a looser and more porous fouling layer according to optical coherence tomography observation. Consequently, the latter was more effective in reducing fouling resistance (33.41% reduction) compared to backwashing by chlorinated-water (8.6%). These findings reveal the critical roles of biofouling in pollutants removal in addition to membrane permeability, which has important implications for addressing seasonal ammonia pollution. (C) 2018 Published by Elsevier B.V.