In a collaborative project, Pennsylvania’s Ohio River basin streams with rich freshwater mussel communities (reference) and depauperate communities (defaunated) were assessed for their mussel stocking potential. The survival and growth of hatchery-reared juvenile Lampsilis cardium in 13 study streams was evaluated. The juveniles were placed in flow through silos from July to September 2021 in seven reference and six defaunated streams. The streams with high survival and growth included those classified as defaunated, Mahoning River, Beaver River, Kiskiminetas River, and South Fork Tenmile Creek, and reference streams, French Creek, Allegheny River, Mahoning Creek and Sandy Creek. Reference streams with colder water temperatures, Tionesta Creek and Clarion River, had high survival but slower growth. Streams with low juvenile survival of included two reference streams, Little Mahoning Creek and Shenango River, and a defaunated stream, Dunkard Creek. A combination of low flow and high stream temperatures in Little Mahoning Creek likely led to the poor outcome for juveniles. In Shenango River and Dunkard Creek follow-up silo studies will be useful to determine why these streams suffered lower than anticipated survival. This is the first Pennsylvania study to use hatchery-raised juveniles placed into silos as a tool to help determine mussel recovery potential and several watersheds are identified that have stocking potential.