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Abstract: Biological indicators, particularly benthic macroinvertebrates, are widely used and effective measures of the impact of urbanization on stream ecosystems. A multimetric biological index of urbanization was developed using a large benthic macroinvertebrate dataset (n = 1,835) from the Baltimore, Maryland, metropolitan area and then validated with datasets from Cleveland, Ohio (n = 79); San Jose, California (n = 85); and a different subset of the Baltimore data (n = 85). The biological metrics used to develop the multimetric index were selected using several criteria and were required to represent ecological attributes of macroinvertebrate assemblages including taxonomic composition and richness (number...