A membrane-aerated biofilm reactor (MABR) was studied for the treatment of wastewater containing acetonitrile, a typical organonitrile compound. The MABR used hydrophobic hollow fiber membranes as the diffusers for bubbleless aeration as well as the carriers for biofilm growth. The objectives were to prevent the stripping-loss of acetonitrile during aeration and to achieve acetonitrile biodegradation plus nitrogen removal simultaneously in a single biolfilm on the membranes. In the MABR, oxygen and substrates were supplied to the biofilm from opposite sides, in contrast to those from the same side in conventional biofilm bioreactors. Operational factors, including surface loading rate and upflow fluid velocity in the bioreactor, on the effect of acetonitrile biodegradation performance were examined. The profiles of dissolved oxygen concentration and microbial activities and populations in the biofilm were investigated. Experimental results showed that, with the adapted microorganisms, removal of acetonitrile at approximately 98.6 and 83.3%, in terms of total organic carbon and total nitrogen, were achieved at a surface loading rate (in terms of membrane surface) of up to 11.29 g acetonitrile/ m 2 · d with an upflow fluid velocity of 12 cm/s and a hydraulic retention time of 30 h. The biofilm on the membranes developed an average thickness of about 1.6 mm in the steady state and consisted of oxic/anoxic/anaerobic zones that provided different functions for acetonitrile degradation, nitrification, and denitrification. The acetonitrile-degrading bacteria in the MABR appeared to secrete more extracellular polymeric substances that enhanced the attachment and development of the biofilm on the membranes. The study demonstrated the potential of using the MABR for the treatment of organonitrile wastewater.