Freshwater ecosystems are facing a deepening biodiversity crisis. Developing robust indicators to assess ecological integrity across large spatial scales and identifying the specific threats and pathways of impairment are thus critically needed if we are to inform freshwater conservation strategies. Here we present the first comprehensive threat assessment across the Colorado River Basin – one of the largest and most endangered river basins in North America – using a spatial framework accounting for the wide range of human activities (land uses, transportation infrastructure, exploitative activities, water withdrawals), pathways (local footprint, overland runoff, upstream cumulative effects), and spatial extent of influence (valley bottom, catchment and river network) known to affect the ecological integrity of riverine ecosystems. We quantified and mapped 69 individual threat indices with geospatial tools for each permanent, ephemeral, and intermittent stream segment within the Basin, encompassing a total of >1,067,700 river kilometers. We further aggregated these indices into components of water quality (diffuse and point-source pollution), hydrology (flow regulation/uses and climate change), and physical system (connectivity and geomorphology). To demonstrate the potential of our framework to inform spatial planning decision processes, we examined the typical combinations of threats experienced by different hydrologic areas and stream segment types, identified candidate watersheds for habitat restoration and enhancement where hotspots of biodiversity and threat overlapped, and assessed the associations between threat indices and in situ measurements of ecological integrity describing a suite of biological (benthic macroinvertebrate, fish), chemical (total nitrogen load, water conductivity), hydrological (flow alteration) and physical indicators (streambed stability, instream habitat complexity). Our assessment highlights clear disparities in term of overall degree of threat that result from different combinations and contributions of individual stressors, with different priorities emerging for perennial versus intermittent or ephemeral stream segments, and between the upper and lower parts of the Basin. Importantly, we showed that our threat indices were generally correlated with biological, chemical, hydrological and physical indicators of ecological integrity they were intended to capture. In addition to its implications for the conservation and management of the highly imperiled Colorado River Basin, our case study illustrates how multi-faceted threat mapping can be used to assess the ecological integrity of riverine ecosystems in the absence of spatially extensive in situ measurements.
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