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Expansion and intensification of human land uses has resulted in the loss of all but remnant areas of natural cover over increasingly vast areas of land. Effective stream assessment in these regions thus requires indicators sensitive to ecosystem change in systems that are already highly stressed. The goal of this study was to determine if benthic macroinvertebrate communities in streams exposed to extensive anthropogenic stress are associated with land-use variables and demonstrate potential to be used as bioindicators for assessment of further land use modification. Using the Grand River in southern Ontario as our model, we sampled for benthic macroinvertebrates in 70 small (3–20 km 2 ) sub-watersheds that comprised a gradient of intensive land use (65–100% developed). Redundancy analysis and variance partitioning results indicated that macroinvertebrate taxonomy and traits were only weakly associated with environmental gradients with the strongest associations occurring at the reach scale. With strong anthropogenic filters present across the watershed, it is likely that the majority of benthic macroinvertebrate communities in the Grand River have experienced a homogenization towards tolerant taxa and traits that respond more to local habitat variables. Threshold Indicator Taxa Analysis (TITAN) revealed 19 valid taxa indicators and 32 valid trait indicators for one or more of six environmental variables (percent sub-watershed agriculture, percent buffer agriculture, pool substrate, sediment reduction, natural channel, and riparian width). However, individual taxon and community change points exhibited broad quantile scores indicating a gradual or random response to environmental variables. Comparison of traits and taxonomic associations with environmental variables showed minimal difference suggesting traits may not offer increased sensitivity to land-use stress. However, several traits did show potential to serve as diagnostic indicators for specific stressors associated with agricultural land use. Our ability to identify even a few indicators in a region with intensive human activity is promising for land managers charged with detecting biotic alterations in this stressed system. Further research developing indicators sensitive to intensification of anthropogenic stress in extensively developed landscapes is critical to ensuring managers have the tools needed to protect and enhance aquatic resources. [ABSTRACT FROM AUTHOR]

• Cumulative effects of human activities are widespread in developed regions. • We assessed effects of agriculture and sewage effluent on stream communities. • Macroinvertebrate and diatom assemblage conditions declined with nutrient enrichment. • Enrichment effects were conditional on upstream nutrient status. • Assessment design needs to account for complexities in multi-use catchments. Human impacts on stream ecosystems are expected to intensify with population growth and climate change. Decisive information on how stream communities respond to cumulative human impacts is therefore integral for protecting streams draining multi-use catchments. To determine cumulative influences of nutrient enrichment and assess more nuanced approaches for the evaluation of human impacts, we present results from one factorial and two gradient assessment designs applied to benthic algae and macroinvertebrate data from 14 mid-order streams in southern Ontario, Canada with pre-existing human impacts (i.e., sewage effluent and agriculture). We found that among stream variability in ecological indicators measured downstream of sewage effluent outfalls confounded our generalized factorial assessment and provided inconclusive information on a known human impact. Despite our gradient assessment also not having strong statistical support, accounting for the extent of nutrient enrichment associated with differences in sewage effluent and agricultural inputs revealed that larger longitudinal changes in stream communities were associated with increased nutrient enrichment. However, re-weighting our nutrient enrichment gradient based on upstream nutrient concentrations to account for nonlinearities in the response of stream communities to nutrient enrichment produced more robust assessment results that were consistent with predicted effects of nutrients on stream ecosystems. Thus, while our factorial assessment suggests that the communities are resistant to nutrients from cumulative human impacts, our targeted gradient assessment demonstrates that the effects of nutrient enrichment are highly conditional on upstream ecosystem conditions. Future assessments may need to go beyond traditional approaches (i.e., impact presence/absence) and more explicitly consider the environmental stressors and their associated complexities related to the impact under investigation. [ABSTRACT FROM AUTHOR]