【Background】As pesticide formulations and dosages increase, acute toxicity tests are importment to find out their effects on non-target organisms for developing environmentally safe application methods. 【Method】The acute toxicity of emamectin benzoate, abamectin, metaflumizone, spirodiclofen, spirotetramat, azoxystrobin, difenoconazole azole, tebuconazole and 2,4-D dimethylamine salt to Bombyx mori and Eisenia fetida were tested, and the toxicity grades were evaluated according to the accepted standard protocol. 【Result】The median lethal concentration after exposure for 96 h of emamectin benzoate, abamectin, metaflumizone, spirodiclofen, spirotetramat, azoxystrobin, difenoconazole azole, tebuconazole and 2,4-D dimethylamine salt to B. mori were 2.05×10-3 a.i.mg·L-1, 8.59×10-4 a.i.mg·L-1, 2.79 a.i.mg·L-1, 250.48 a.i.mg·L-1, 11.52 a.i.mg·L-1, 272.18 a.i.mg·L-1, 2.50 a.i.mg·L-1, 1.93×10-2 a.i.mg·L-1 and 534.47 a.i.mg·L-1 respectively. The 14 days median lethal concentration to E. fetida were 11.05 a.i.mg·kg-1dw, 6.29 a.i.mg·kg-1dw, >100 a.i.mg·kg-1dw, >100 a.i.mg·kg-1dw, >100 a.i.mg·kg-1dw, >100 a.i.mg·kg-1dw, 99.13 a.i.mg·kg-1dw, 115.31 a.i.mg·kg-1dw, >100 a.i.mg·kg-1dw. Emamectin benzoate, abamectin and tebuconazole were severely toxic to B. mori, and metaflumizone, spirotetramat and difenoconazole had high toxicity, only azoxystrobin toxicity was low. Abamectin was moderately toxic to E. fetida, all other tested pesticides had low toxicity. 【Conclusion and significance】Significant differences between toxicity of pesticides to B. mori and E. fetida were found, which can serve as reference for the regulation and environmental protection.