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Defining stream reference conditions is integral to providing benchmarks to ecological perturbation. We quantified channel geometry, hydrologic and environmental variables, and macroinvertebrates in 62 low‐gradient, SE United States (U.S.) Sand Hills (Level IV ecoregion) sand‐bed streams. To identify hydrogeomorphic reference condition (HGM), we clustered channel geometry deviation from expectations given watershed area (Aws), resulting in two HGM groups discriminated by area at the top of bank (Atob) residuals <0.6 m2 and >0.6 m2 predicted to be HGM reference/nonreference streams, respectively. Two independent partial least squares discriminate analyses used (1) hydrologic/environmental variables and (2) macroinvertebrate mean trait values (mT) on 10 reference/nonreference stream pairs of similar Aws for classification validation. Nonreference streams had flashier hydrographs and altered flow magnitudes, lower organic matter, coarser substrate, higher pH/specific conductivity compared with reference streams. Macroinvertebrate assemblages corresponded to HGM groupings, with mT indicative of multivoltinism, collector‐gatherer functional feeding groups, fast current‐preference taxa, and lower Ephemeroptera, Plecoptera, and Trichoptera richness and biotic integrity in nonreference streams. HGM classifications in Sand Hills, sand‐bed streams were determined from channel geometry. This easily implemented classification is indicative of contemporary hydrologic disturbance resulting in contrasting macroinvertebrate assemblages.  相似文献   
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