Recent media attention regarding microplastic pollution in the Tennessee River has highlighted the paucity of information regarding microplastic pollution and its effects on freshwater organisms. We developed a pilot study to quantify and compare microplastics ingestion by freshwater macroinvertebrates in urban and rural streams of the Upper Tennessee River Basin. We collected Tanback Crayfish Cambarus girardianus and Asian Clam Corbicula fluminea specimens from Third Creek in Knoxville, Tennessee, and Little River in Walland, Tennessee and dissolved all organic animal tissues using alkaline digestion. We examined the remaining filtrate under a microscope, counted the microplastics and listed their characteristics with respect to type, color and shape. We evaluated 25 crayfish and 7 clams. The highest number of microplastics found in a crayfish was 12 and the highest in a clam was 10. Both were from the Little River. The average number of microplastics found in specimens from Third Creek was 4.625 and the average from the Little River was 5.0. Overall, we found microplastics were higher in specimens from the Little River, but the range was similar to Third Creek. Most microplastics we identified we elongated fibers. Most contemporary studies regarding microplastics ingestion by animals have been focused on marine ecosystems. These results of our study, while somewhat inconclusive, demonstrate that microplastics ingestion by macroinvertebrates in freshwater ecosystems demands similar attention.