Abstract Environmental factors act at multiple spatial scales in a hierarchical manner to shape the organization of biota. However, the relative influence of different scale-related factors is poorly known, especially in Atlantic Forest Blackwater streams. Therefore, we herein aimed to evaluate local, landscape and spatial factors that shape fish assemblages in 14 blackwater restinga coastal Atlantic Forest streams under natural conditions and verify species occurrence patterns among four sub-basins during the low-precipitation season. When we combined local, landscape and spatial factors, variance partitioning explained a high proportion of variation in species matrix. Local variables pH and Total Dissolved Solids explained most of the variability, and these were the most important factors in determining fish community structure. Significant differences in fish assemblage structure among the four sub-basins were observed, and Mimagoniates microlepis, Phalloceros harpagos, and Hollandichthys multifasciatus were the species that most contributed to this dissimilarity. The important contribution of local predictors, the high number of endemic species herein recorded, the presence of an endangered species (Spintherobolus broccae), and near pristine conditions, may be used as baseline conditions for the assessment of similar environments.
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