The amount of ultraviolet radiation (UVR) reaching freshwater organisms is predicted to increase with climate change. Exposure to UVR causes harmful DNA damage in amphibian larvae. This damage is repaired by enzymes. Cool temperatures slow DNA repair rates, compounding the negative impacts of UVR exposure. It is unknown whether amphibian species differ in the thermal sensitivity of UV-induced DNA damage or if larvae can acclimate to low temperatures to compensate for the effects of temperature on DNA repair. We reared larvae of three closely related Limnodynastid amphibian species and exposed them to a high UVBR dose (80 µW cm-2 for 2 hours) at a range of temperatures and sampled over the following 24 h period while UV-damaged DNA was repaired. Cool temperatures increased DNA damage in all three species. However, the magnitude of DNA damage incurred, the rates of DNA repair, and the thermal sensitivity of DNA repair rates differed between species. Striped marsh frog larvae were also acclimated to cooler or warmer temperatures prior to an acute high UVBR exposure. These larvae reared at low temperature had lower DNA damage, demonstrating capacity for acclimation to UV and temperature likely facilitated through physiological colour changes. Importantly, laboratory studies on UVR may not capture ecologically realistic UVR levels in freshwater ecosystems. We monitored UV index and temperature using a variety of methods to assess the ecological relevance of laboratory investigations into UVR effects on freshwater organisms within a local context. We found significant capacity for high UVR exposure and in amphibian habitats, but patterns of UV exposure dose and intensities were highly variable and not always reflective of laboratory-based studies. Our findings highlighted the need to carefully consider species-specific physiology, phenotypic plasticity, and ecological relevance when forecasting the effects of changing UVR on freshwater organismal physiology.