1. Protected area networks for river ecosystems must account for the highly connected nature of river habitats and the fact that conditions in distant locations can influence downstream habitats and biota. We used Marxan conservation planning software to address the unique constraints of reserve design in river ecosystems and structure a reserve network to overcome key challenges to freshwater conservation.2. The range limits of 63 fish species in Mesoamerica were predicted and used in Marxan to design a network of conservation focal areas that encompasses 15% of the range of each species in areas with low risk of environmental degradation. Upstream risk intensity was estimated by propagating landscape-based sources of stress downstream alo...