In Europe, spreading organic wastes to fertilize soils is an alternative commonly used instead of chemical fertilizers. Through their contributions of nutrients and organic matter, these wastes promote plant growth and thus agricultural production. However, these organic amendments can also contain mineral and organic pollutants requiring chemical and ecotoxicological analyses to guarantee their harmlessness on soil and its organisms during spreading. The purpose of this study was to assess the potential toxicity of three organic amendments from diferent sources (sewage sludge, dairy cow manure, dairy cow slurry) by performing chemical analyses and acute toxicity tests on three types of organism: earthworms, plants, soil microbial communities. Chemical analysis revealed a higher content of certain pharmaceuticals, polycyclic aromatic hydrocarbons and metals in sewage sludge in comparison with the two other types of organic wastes. The ecotoxicological assessment showed a dose-dependent efect on soil organisms for the three organic amendments with higher toxic efects during the exposure tests with a soil amended with dairy cow slurry. However, at realistic spreading doses (10 and 20 g kg−1 dry weight of organic amendments) on a representative exposed soil, organic amendment did not show any toxicity in the three organisms studied and had positive efects such as increased earthworm biomass, increased plant root growth and earthworm behavior showing attraction for organic amendment. On the contrary, exposure assays carried out on a limited substrate like sandy soil showed increased toxicity of organic amendments on plant germination and root growth. Overall,bthe ecotoxicological analysis revealed greater toxicity for soil organisms during the amendment of cow slurry, contrary to the chemical analysis which showed the potential high risk of spreading sewage sludge due to the presence of a higher quantity of pollutants. The analysis of the chemical composition and use of acute toxicity tests is the frst essential step for assessing the ecotoxicological risk of spreading organic amendments on soil organisms. In addition to standard tests, the study suggests using a representative soil in acute toxicity tests to avoid overestimating the toxic efects of these organic amendments.