We tested macroinvertebrate assemblages collected from 1979–2015 for temporal variation in structure and for impacts of the Clean Water Act of 1974. Collections were at ten sites on the mainstem of the West Fork White River. We used family-level taxonomy for macroinvertebrates that resulted in 77 families and 92,477 individuals. Macroinvertebrate families were further classified by trophic and tolerance traits and tested for temporal variation. We defined river reaches as upstream, urban, and downstream of Muncie, Indiana for analyses. Taxonomic richness increased over the study. A nonmetric multidimensional scaling (NMDS) analysis identified high temporal variation as assemblage structure differed among decades. Spatial analyses using NMDS indicated significant differences by river location upstream, urban, and downstream. NMDS and Analysis of Similarities (ANOSIM) by trophic relationship and tolerance values did not result in significant temporal or spatial patterns. Our results show the macroinvertebrate assemblages of the West Fork White River improved, likely due to implementation of the Clean Water Act.