Shenandoah National Park contains hundreds of natural springs which help form the headwaters of three major watersheds that empty into Chesapeake Bay. Virginia Tech partnered with the National Park Service Inventory and Monitoring program to sample macroinvertebrates and environmental measures related to water quality, habitat availability, and human use at 63 springs within the park, of which, 59 could were assessed and analyzed. The goals of this inventory were to assess the distributions of macroinvertebrates across springs in the park. Additionally, we sought to determine whether macroinvertebrate diversity and the presence of rare taxa have been impacted by modifications made to springs and/or by visitor use. We identified 205 unique taxa across the 59 springs. Of those taxa, we identified 17 rare taxa using NatureServe’s Conservation Status Rank. Rare taxa were present in 32 springs, 19 of which were modified and 13 were unmodified. Class Insecta represented 90% of rare taxa that were mostly in orders Trichoptera, Plecoptera, Ephemeroptera and Odonata. There was no difference in total taxa richness or abundance between the spring types. Richness and abundance for all taxa and for rare taxa were greater, on average, in modified springs; however, these differences were not statistically significant. The diversity of spring types, locations and habitats appear to support a diversity of species that are not being negatively affected by visitor usage or spring box infrastructure. The springs provide a regional species pool at high density that are likely source populations for the headwater streams.