BACKGROUND: Inappropriate discharge of wastewaters and industrial effluents are becoming detrimental to the aquatic environment. The presence of toxic substances on wastewaters can be detected by physicochemical and biological methods. However, physicochemical methods do not give any information about biological toxicity. Therefore, in this study we tried to detect the presence of toxic substance on waters using sulfur-oxidizing bacteria (SOB) as a bioassay. MATERIALS AND RESULTS: The SOB biosensor was first stabilized using synthetic stream water and operated in both continuous and semi-continuous mode. When the SOB biosensor was operated in continuous mode, the effluent electrical conductivity (EC) stabilized at~1.72 dS/m. While in the case of semi-continuous, the EC stabilized at~0.6 dS/m. The SOB system was also operated at different reaction times to ascertain the shortest reaction time for monitoring the toxicity. Finally, the SOB biosensor was fed with nitrite as toxic substance. When 5 mg/L of nitrite was added to the SOB system, the EC decreased immediately. However, the EC recovered after few cycle. CONCLUSION: This study shows that the SOB biosensor can be used as warning system to protect aquatic environment from hazardous materials. Although SOB biosensor can not give specific information about the toxic substances, it can assess whether the water is toxic or not.