Environmental impacts from ecotourism and outdoor recreation activities on terrestrial and aquatic ecosystems are well-reported in the literature, but less is known regarding the impacts of outdoor environmental education activities. Student activity during stream classes may cause substrate disruption and localized impacts on stream macroinvertebrates. We hypothesized that student activity would negatively impact macroinvertebrate community structure in three habitat types (riffle, run, pool) within a site regularly used for stream classes while no impact from student activity would occur in the same three habitat types within an unused site. We addressed the hypothesis by sampling macroinvertebrates monthly for one year in the riffles, runs, and pools at the class site and the unused site within a fourth order stream in central Ohio. The results indicated reduced macroinvertebrate abundance and richness in the riffle at the class site during periods with student activity and no differences between site types during periods without student activity. No impacts of stream classes on macroinvertebrate communities were observed in runs or pools. The results suggest that environmental education organizations should avoid repetitive use of the same site for their stream classes to avoid reductions of macroinvertebrate abundance and taxa richness that can impact the students’ educational experience.