Macroinvertebrate assemblages and environmental variables were evaluated at 45 stream sites throughout the Puget Sound Basin, Washington, USA. Environmental variables were measured at 3 spatial scales: reach, local, and whole watershed. Macroinvertebrate distributions were related to environmental variables using canonical correspondence analysis to determine which variables and spatial scales best explained the observed community composition and to identify biota-land cover optima. The calculation of a biota-land cover optimum was a 2-step process. First, an individual taxon's optimum was estimated for a particular land cover by weighting the mean value for that land cover by the abundance of that taxon at all sites. Second, the biota-land cover optimum was determined as the point at which the greatest numbers of taxa, at their calculated optima, appeared for a particular land cover. Sampling reaches were located on streams in watersheds with varying levels of forest, agriculture, and urban/suburban land cover that represented the full range of physical conditions typically found in Puget Sound streams. At the reach scale, taxa composition was correlated with conductivity and mean velocity. At the local and whole-watershed scales, taxa composition was correlated with % forest and agricultural land cover and % forest and bedrock land cover, respectively. For all of the scales, the dominant environmental variables represented an anthropogenic gradient. There was little difference in the amount of variability explained by each spatial scale. At the local-watershed scale, a biota-land cover optimum of ???80 to 90% forest land cover was identified. The total number of taxa at their optima declined rapidly as forest land cover within the local scale declined below 80 to 90%. At the whole-watershed scale, a biota-land cover optimum of 70 to 80% forest land cover was identified. The total number of taxa at their optima declined rapidly as forest land cover within the whole watershed declined below 70 to 80%. Our results suggest that macroinvertebrates can be used both as quantitative indicators of environmental conditions at multiple scales and indicators of land cover optima. Further examination of these optima could be used to establish priorities for conservation and restoration efforts.