Microbial cultures originated from an oil reservoir were used in three biofilm reactors and effects of sulphide and nitrate loading rates and molar loading ratio on the removal of sulphide, nitrate and acetate, and composition of end products were investigated. Application of biofilms improved sulphide and nitrate removal rates significantly when compared with freely suspended cells. Maximum sulphide and nitrate removal rates under autotrophic conditions were 30.0 and 24.4 mM h(-1) respectively (residence time: 0.5 h). Oxidation of acetate occurred only at nitrate to sulphide molar loading ratios around 0.7 or higher when nitrate was present at levels higher than that required for oxidation of sulphide to sulphur. Conversion of sulphide to sulphate increased from 0% to 66% as nitrate to sulphide molar loading ratio was increased from 0.34 to 3.98. The highest nitrate and acetate removal rates in the bioreactor operated under heterotrophic conditions were 183.2 and 88.0 mM h(-1), respectively (residence time: 0.8 h). (c) 2010 Elsevier Ltd. All rights reserved.