Oxygen injection is often used to control biogenic production of hydrogen sulfide in sewers. Experiments were carried out on a laboratory system mimicking a rising main to investigate the impact of oxygen injection on anaerobic sewer biofilm activities. Oxygen injection (15-25mg O(2)/L per pump event) to the inlet of the system decreased the overall sulfide discharge levels by 65%. Oxygen was an effective chemical and biological oxidant of sulfide but did not cause a cessation in sulfide production, which continued in the deeper layers of the biofilm irrespective of the oxygen concentration in the bulk. Sulfide accumulation resumed instantaneously on depletion of the oxygen. Oxygen did not exhibit any toxic effect on sulfate reducing bacteria (SRB) in the biofilm. It further stimulated SRB growth and increased SRB activity in downstream biofilms due to increased availability of sulfate at these locations as the result of oxic conditions upstream. The oxygen uptake rate of the system increased with repeated exposure to oxygen, with concomitant consumption of organic carbon in the wastewater. These results suggest that optimization of oxygen injection is necessary for maximum effectiveness in controlling sulfide concentrations in sewers.