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Agricultural land use is a primary driver of environmental impacts on streams. However, the causal processes that shape these impacts operate through multiple pathways and at several spatial scales. This complexity undermines the development of more effective management approaches, and illustrates the need for more in‐depth studies to assess the mechanisms that determine changes in stream biodiversity. Here we present results of the most comprehensive multi‐scale assessment of the biological condition of streams in the Amazon to date, examining functional responses of fish assemblages to land use. We sampled fish assemblages from two large human‐modified regions, and characterized stream conditions by physical habitat attributes and key landscape‐change variables, including density of road crossings (i.e. riverscape fragmentation), deforestation, and agricultural intensification. Fish species …