To compensate the problems of chemical assay in detoxification of recalcitrant and a practical approach in selection of bioremediation bacteria, a simple and rapid toxicity evaluation system was constructed based on Lumbricus rubellus. Long term-culture and specific equipment are not necessary, and semi-quantitative analysis of toxicity at sub-lethal concentration is possible by measuring of dose-dependent increased yellowish secreted compounds. When the toxicity of endosulfan, its metabolites and structurally related chemicals were measured for 24 h, the results were coincided with previous reports. Toxicity was found in endosulfan, endosulfan sulfate, aldrin, and dieldrin, respectively. Rapid and economic selection of endosulfan-detoxifying bacteria was possible using our system. Klebsiella pneumoniae KE-1, K. oxytoca KE-8 and Pseudomonas sp. KS-2P, reported endosulfan degrading bacteria, ameliorated the endosulfan toxicity, whereas E. coli, B. subtilis and other bacteria failed to protect the toxicity of endosulfan in L. rubellus. Our results suggest that the constructed system is useful to selection of microorganism as well as toxicity evaluation against toxic recalcitrants.