Subsets of nucleus accumbens (NAc) neurons process information about operant responses for drugs as well as natural rewards (food and water) by excitations and inhibitions in firing rate time-locked to the operant response. The degree to which ensembles of neurons exhibit similar firing patterns when encoding cues and operant responses across different reinforcer conditions will provide critical information regarding the functional organization of this nucleus. The present experiment evaluated the relative contribution of subsets of accumbens neurons that encode distinct features of lever press responding for ethanol vs. water. Electrophysiological recordings (n = 153 neurons) were made in the accumbens of rats trained on concurrent reinforcement schedules for ethanol and water throughout a self-administration session. During operant responding, 52% of neurons exhibited patterned discharges characterized by significant increases or decreases in firing rate of ± 1 s relative to lever presses for ethanol and/or water. Of these phasic cells, 85% discriminated between presses for ethanol and water (i.e. exhibited firing patterns unique to one reinforcer type), while 15% exhibited identical firing patterns relative to lever presses for both reinforcers. Notably, the data revealed that both high ethanol preference and spatially distinct lever positions contributed to the reinforcer specificity. Together, these data demonstrate that subsets of NAc neurons encode conditioned and instrumental aspects of ethanol vs. water reinforcement in well-trained rats, and that reinforcer preference and spatial cues are important components of this differential information processing.