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Anthropogenic ozone depletion has led to a 2–5% increase in ultraviolet B radiation (UVBR) levels reaching the earth's surface. Exposure to UVBR causes harmful DNA damage in amphibians, but this is minimized by DNA repair enzymes such as thermally sensitive cyclobutane pyrimidine dimer (CPD)-photolyase, with cool temperatures slowing repair rates. It is unknown whether amphibian species differ in the repair response to a given dose of UVBR across temperatures. We reared larvae of three species ( Limnodynastes peronii , Limnodynastes tasmaniensis and Platyplectrum ornatum ) at 25°C and acutely exposed them to 80 µW cm −2 UVBR for 2 h at either 20°C or 30°C. UVBR-mediated DNA damage was measured as larvae repaired damage in photoreactive light at their exposure temperatures. Cool temperatures increased DNA damage in two species and slowed DNA repair rate in P. ornatum . The magnitude of DNA damage incurred from UVBR was species-specific. Platyplectrum ornatum had the lowest CPDs and DNA repair rates, and the depressive effects of low temperature on photorepair were greater in L. tasmaniensis . Considering the susceptibility of most aquatic organisms to UVBR, this research highlighted a need to consider the complexity of species-specific physiology when forecasting the influence of changing UVBR and temperature in aquatic ecosystems.