Acute toxic effects of three typical organic pollutants 1 ,2,4-trichlorobenzene (1,2,4-TCB), nitrobenzene and chlorpyrifos were investigated using Tetrahymena thermophila and Limnodrilus hoffmeisteri as living test organisms under laboratory conditions. The results showed that with the increase of pollutants' concentration and the extension of time, toxicity of the three kinds of pollutants significantly enhanced, and the mortality of two kinds of aquatic organisms also had a rising trend, and an obvious dose-effect relationship. The 96 h-LC50. values of 1 ,2, 4-TCB, nitrobenzene and chlorpyrifos were 71.88, 285.76, and 5.50 mg x L(-1) for L. hoffmeisteri and 15.58, 140.22, and 14.69 mg x L(-1) for T. thermophila. These results showed that the toxicity among the three typical pollutants to T. thermophila was 1 , 2,4-TCB > chlorpyrifos > nitrobenzene. Findings were able to provide more information on water quality criteria and more data on their toxicity to indigenous aquatic organisms in China.