BackgroundUrban areas are often built along large rivers and surrounded by agricultural land. This may lead to small tributary streams that have agricultural headwaters and urbanized lower reaches. Our study objectives assessed are as follows: (1) landscape, geomorphic, and water quality variables that best explained variation in aquatic communities and their integrity in a stream system following this agricultural-to-urban land use gradient; (2) ways this land use gradient caused aquatic communities to differ from what would be expected for an idealized natural stream or other longitudinal gradients; and (3) whether the impacts of this land use gradient on aquatic communities would grow larger in a downstream direction through the agricultural and urban developments. Our study area was an impaired coldwater stream in Michigan, USA.ResultsMany factors structured the biological communities along the agricultural-to-urban land use gradient. Instream woody debris had the strongest relationship with EPT (Ephemeroptera, Plecoptera, and Trichoptera) abundance and richness and were most common in the lower, urbanized watershed. Fine streambed substrate had the strongest relationship with Diptera taxa and surface air breather macroinvertebrates and was dominant in agricultural headwaters. Fish community assemblage was influenced largely by stream flow and temperature regimes, while poor fish community integrity in lower urban reaches could be impacted by geomorphology and episodic urban pollution events. Scraping macroinvertebrates were most abundant in deforested, first-order agricultural headwaters, while EPT macroinvertebrate richness was the highest downstream of agricultural areas within the urban zone that had extensive forest buffers.ConclusionEnvironmental variables and aquatic communities would often not conform with what we would expect from an idealized natural stream. EPT richness improved downstream of agricultural areas. This shows promise for the recovery of aquatic systems using well-planned management in watersheds with this agricultural-to-urban land use pattern. Small patches of forest can be the key to conserving aquatic biodiversity in urbanized landscapes. These findings are valuable to an international audience of researchers and water resource managers who study stream systems following this common agricultural-to-urban land use gradient, the ecological communities of which may not conform with what is generally known about land use impacts to streams.