Throughout the canyons of the Colorado and Uncompahgre Plateaus, water is a limited resource for wildlife, with patchy distribution and seasonal availability. Tributary creeks within these canyons drain into mainstem rivers, providing habitat and breeding sites for native amphibians. Yet, little is known about the diversity and distribution of amphibians that live in these harsh, dynamic environments. In addition, the rivers that border these canyon tributaries may serve as corridors for nonnative species and disease. The American Bullfrog (Lithobates catesbeianus) is a nonnative species in western Colorado known to prey on native amphibians and act as a reservoir for pathogens such as Batrachochytrium dendrobatidis (Bd). From 2019 to 2022, we surveyed for amphibians using visual encounter surveys (VES) and environmental DNA (eDNA) surveys throughout the McInnis Canyons National Conservation Area (MCNCA), the Dominguez–Escalante National Conservation Area (DENCA), and the Dolores River Canyon Wilderness Study Area (DRCWSA). Our primary goals were to document the diversity and distribution of native amphibians in the canyonlands and evaluate potential threats to these species from bullfrogs and Bd. We confirmed that sensitive species, such as the Great Basin Spadefoot (Spea intermontana) and the Northern Leopard Frog (Lithobates pipiens), inhabit these protected areas. In most cases, bullfrogs were not detected within ephemeral tributaries, but bullfrog DNA was detected in some tributaries at the confluence with the mainstem rivers. In Mee Canyon (MCNCA), however, bullfrogs were found within the tributary, up to 3 km from the Colorado River. A bullfrog individual removed from this canyon tested positive for Bd, and diet contents suggested that native amphibians are potential prey in this system. Nonnative predators and disease pose a threat to native amphibians, alongside environmental changes such as drought and hydrological shifts driven by ongoing climate change.