Resumo. The present study aimed to evaluate the production of seedlings of Mimosa setosa from the use of different combinations of substrates, namely, cattle manure, poultry litter, quail manure and commercial substrate. The experiment was conducted in the Nursery Center for Agricultural Sciences, at the Federal University of Espírito Santo state, and mounted in a completely randomized design (CRD). When the seedlings reached five months after sowing, the following characteristics were measured: total height, collar diameter, plant height/collar diameter ratio, dry mass of shoot, dry mass of root, total dry mass and dry mass ratio of shoot/ root dry mass, along with the Dickson quality index (IQD). It was found that, for the production of seedlings of Mimosa setosa, it is recommended a formulation consisted by 25% of commercial substrate + 35% of poultry litter + 40% of subsoil, having this one the best averages in morphological characteristics total height, collar diameter, plant height/collar diameter ratio, dry mass of shoot, dry mass of root and total dry mass.