The optimal operation conditions in an anoxic sulfide oxidizing (ASO) bioreactor were investigated. The maximal loading rates (LRs) observed through decreasing hydraulic retention time (HRT) at fixed substrate concentration are higher than those by increasing substrate concentration at fixed HRT. The sulfide oxidation in ASO reactor was partially producing both sulfate and sulfur; but the amount of sulfate produced was approximately one third that of sulfur. The process was able to tolerate high sulfide concentration, as the sulfide removal percentage always remained near 99% when influent concentration was up to 580 mg/L. It tolerated relatively lower nitrate concentration because the removal percentage dropped to 85% when influent concentration was increased above 110 mg/L. The process can tolerate shorter HRT but careful operation is needed. Nitrate conversion was more sensitive to HRT than sulfide conversion since the process performance deteriorated abruptly when HRT was decreased from 3.12 h to 2.88 h. In order to avoid nitrite accumulation in the reactor, the influent sulfide and nitrate concentrations should be kept at 280 mg/L and 67.5 mg/L respectively. Present biotechnology is useful for removing sulfides from sewers and crude oil.