Living in a multi-stressor world: nitrate pollution and thermal stress interact to affect amphibian larvae.

The interaction of widespread stressors such as nitrate pollution and increasing temperatures associated with climate change are likely to affect aquatic ectotherms such as amphibians. The metamorphic and physiological traits of amphibian larvae during the critical onset of metamorphosis are particularly susceptible to these stressors. We used a crossed experimental design subjecting Rana temporaria larvae to four constant rearing temperatures (18, 22, 26, 28 °C) crossed with three environmentally relevant nitrate concentrations (0, 50, 100 mg×L-1) to investigate the interactive and individual effects of these stressors on metamorphic (i.e., growth and development) and physiological traits (i.e., metabolism and heat tolerance) at the onset of metamorphosis. Larvae exposed to elevated nitrate concentrations and thermal stress displayed increased metabolic rates but decreased developmental rate, highlighting interactive effects of these stressors. However, nitrate pollution alone had no effect on either metamorphic or physiological traits, suggesting that detoxification processes were sufficient to maintain homeostasis but not in combination with increased rearing temperatures. Furthermore, larvae exposed to nitrate displayed diminished abilities to exhibit temperature-induced plasticity in metamorphosis timing and heat tolerance, as well as reduced acclimation capacity in heat tolerance and an increased thermal sensitivity of metabolic rate to higher temperatures. These results highlight the importance of considering the exposure to multiple stressors when investigating how natural populations respond to global change.