【Background】Propineb is an organic sulfur fungicide with protective broad spectrum but with long residual effect in soil. Being readily available, propineb is widely used for preventing plant diseases. It may have some negative effects on other organisms in the fields. 【Method】Toxicity of propineb on six untargeted species (Japanese quail, bee, silkworm, zebrafish, daphnia and earthworm) was tested by various indoor simulated tests, according to "Experimental Guideline for Environmental Safety Evaluation of Chemical Pesticides", and toxicity bioassay was also conducted. 【Result】The results showed that the toxicity of propineb to bird and bee was low. The LC50 (96 h) for silkworm was 0.3990 mg·L-1 and considered extremely toxic. The LC50 (96 h) for fish and the EC50 (48 h) for daphnia were considered medium in terms of toxicity with values of 1.4764 mg·L-1 and 2.7406 mg·L-1 respectively. and the LC50 (14 d) of propineb for earthworm was 29.0737 mg·kg-1, representing a low toxicity. 【Conclusion and significance】This study provides scientific evidence for risk assessment and environmental security management of propineb, and argument and scientific guidelines for its reasonable use.