The dataset comprise environmental characterization for 86 streams in the Turvo-Grande and São José dos Dourados basins, Upper Paraná River basin, northwestern of São Paulo State, Brazil. We selected those streams considering the environmental gradient in the region (i.e., pasture-sugarcane transition). In each stream we quantified 31 local descriptors related with marginal vegetation (e.g., marginal grasses, roots, shrubs), water properties (e.g., water temperature, pH), instream structures (e.g., litter, wood debris), stream channel morphology (e.g., width, depth, flow), habitat composition and heterogeneity (e.g., mesohabitat, substrate composition). In addition, for each stream catchment (i.e., catchment area delimited upstream from the sampled stream reach), we measured 30 descriptors related with land-use composition and heterogeneity quantified through (i) the number of the land-use classes (i.e., richness), (ii) diversity of land-use classes (i.e., Shannon index), and (iii) the relative proportion of each land-use class. For a detailed description of how variables were obtained see Molina et al. (2017); Roa-Fuentes &amp; Casatti (2017); Roa-Fuentes et al. (2019, 2020). A data matrix is provided in txt extension and a script with exploratory data analysis in R software (R Development Core Team 2020). <strong>References</strong> <em>- Molina, M. C., C.A. Roa-Fuentes, J.O. Zeni &amp; L. Casatti, 2017. The effects of land use at different spatial scales on instream features in agricultural streams. Limnologica 65: 14–21. </em> <em>- R Development Core Team, 2020. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL http://www.R-project.org/</em> <em>- Roa-Fuentes, C. A. &amp; L. Casatti, 2017. Influence of environmental features at multiple scales and spatial structure on stream fish communities in a tropical agricultural region. Journal of Freshwater Ecology 32: 281–295.</em> <em>- Roa-Fuentes, C. A., J. Heino, M. V. Cianciaruso, S. Ferraz, J. O. Zeni &amp; L. Casatti, 2019. Taxonomic, phylogenetic and functional β-diversity patterns of stream fish in tropical agroecosystems. Freshwater Biology 64: 447–460. </em> - <em> Roa-Fuentes, C.A., Heino, J., Zeni, J.O. et al. Importance of local and landscape variables on multiple facets of stream fish biodiversity in a Neotropical agroecosystem. Hydrobiologia (2020). https://doi.org/10.1007/s10750-020-04396-7 </em>