Lotic systems within the Great Plains are characterized by highly fluctuating conditions through both space and time. Fishes inhabiting these systems have adopted specific life-history strategies to survive in such environments; however, anthropogenic disturbance to prairie streams has resulted in declines and extirpation of many native stream fishes. Terrestrial protected areas (i.e., parks and reserves) are designated to support native flora and fauna and, it is assumed, to provide protection to native fishes. We assessed the presence and relative abundance of stream fish populations within protected areas along the Niobrara River in western Nebraska based on data collected during 1979, 1989, 2008, and 2011. The spatial extent of protection, landscape changes resulting in degraded physiochemical parameters, and introduced species may reduce the effectiveness of these terrestrial protected areas in protecting native fishes in Great Plains stream environments.