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Metsulfuron-methyl is one of the most used sulfonylurea herbicides, being applied alone in pre-emergence and with a mineral oil (as adjuvant) in post-emergence. In risk assessment of pesticides, ecotoxicity tests have been applied to assess the effects of products and mixtures under laboratory conditions, but they are limited in their ecological relevance when compared to field assessments. Considering the differences between laboratory and field exposure, and the lack of data on the effects of metsulfuron-methyl in natural soils, this study consisted in a set of tests to assess the ecotoxicity of this herbicide applied alone, combined with an adjuvant (mineral oil) and the adjuvant applied alone, both under laboratory and field exposure, with artificial and natural soil respectively. Reproduction tests with four non-target soil invertebrates species were performed in laboratory, while two experiments were performed in field evaluating avoidance behaviour, feeding activity, mesofauna abundance and pesticide residual. Laboratory results showed that metsulfuron-methyl alone is not a threat to soil fauna on the recommended doses. However, the presence of mineral oil as adjuvant showed ecotoxicity to Eisenia andrei, Enchytraeus crypticus and Proisotoma minuta on laboratory tests. Field evaluations indicated that metsulfuron-methyl and the adjuvant do not impaired the feeding activity of the soil fauna. The low abundance of native communities could be related to soil management. Results showed that laboratory and field evaluations are necessary to better understanding of the effects of pesticides to soil fauna and adjuvant addition should be considered on pesticides risk assessment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)