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Objectives:The purpose of this study is to evaluate the effects of heavy metals (Zn2+, Pb2+, Hg2+, Cd2+) toxicity in a real-time biosensor which is based on measuring the attenuation of light intensity emitted by Vibrio fischeri. Methods:The inhibition test included four heavy metals and the spiked effluent wastewater. The toxicity of the test samples was assessed by freeze-dried bacteria (Vibrio fischeri NRRL-B-11177). The pH of the samples was adjusted to 7±0.2 before conducting the toxicity tests in oder to eliminate potential pH effects. The results obtained from the real-time biosensor that can be measuring the toxicity of the samples as a function of the emitted light. within a short period time (2 min). And then EC20 and EC50 values was estimated from inhibition curves. Results and Discussion:In case of the inhibition test for dilluents of heavy metals, EC20 values were estimated as < 0.2 mg/L for Zn(II), 0.45 mg/L for Hg(II), 0.58 mg/L for Cd(II) and 1.95 mg/L for Pb(II) and EC50 values were estimated as 0.25 mg/L for Zn(II), 0.5~1.0 mg/L for Hg(II), 1.38 mg/L for Cd(II) and 3.76 mg/L for Pb(II). The sensitivity ranking of heavy metals was in order as Zn(II) > Hg(II), > Cd(II) > Pb(II). In case of the inhibition test for the spiked effluent wastewater, EC20 values were estimated as 0.38 mg/L for Hg(II), 0.58 mg/L for Zn(II), 1.45 mg/L for Pb(II) and 1.95 mg/L for Cd(II) and EC50 values were estimated as 0.53 mg/L for Hg(II), 1.13 mg/L for Zn(II), 6.44 mg/L for Pb(II) and 7.82 mg/L for Cd(II). The sensitivity ranking of the spiked effluent wastewater was in order as Hg(II) > Zn(II) > Pb(II) > Cd(II). Compared to the Emssion Limit Value (ELV) in Korea, the EC20 value of Zn(II) was found to be lower than ELV so that it was suitable for screening of Zn(II) as to meet the criteria of effluent wastewater. However, the other EC20 value of heavy metals can be used to control the unit processes and avoid accidental discharge. Conclusions:The assessment of the toxicity of four heavy metals and spiked samples with them was performed in this study, by real-time biosensor using bioluminescent bacteria. The EC20 value of the spiked sample with Zn(II) has been found to be 0.58 mg/L lower than its ELV so that it was considered as a suitable screening for determining whether or not to exceed the permissible limit value. The use of real-time biosensor is required in combination with physicochemical analyses for an integrated screening of the industrial effluent properties.