Chloramphenicol (CAP) is a frequently detected environmental pollutant. In this study, an electroactive biofilm for CAP reduction was established by initially in the anode and then inverting to the cathode. The established biocathode could enhance the reduction of CAP to the nitro-group reduced CAP (AMCl2) and further dechlorinated form (AMCl), both had lost the antibacterial activity. The phylogenetic diversity of the acclimated biofilm was decreased after the polar inversion. Proportions of functional bacterial genera, including Geobacter, Desulfovibrio and Pseudomonas responsible for the bidirectional electron transfer and nitroaromatics reduction, had increased 28%, 104% and 43% in the cathode. The relatively high abundances (over 50%) of Geobacter in anode and cathode were rarely detected for the nitroaromatics reduction. This study provides new insights into the electroactive biofilm structure improvement by the polarity inversion strategy for refractory antibiotics degradation. (C) 2016 Elsevier Ltd. All rights reserved.