Multiple interacting stressors influence development, growth, and morphology of Pacific Chorus Frog (Pseudacris regilla) larvae

Freshwater ecosystems are threatened by climate change and invasive species. Amphibians are important members of freshwater communities and are susceptible to negative effects of climate change and invasion. Further-more, both climate change and invasion can influence density of amphibian larvae at the microhabitat scale because of larval clustering. To understand the effects of climate change and invasion on Pacific Chorus Frog Pseudacris regilla (Baird and Girard, 1852) larvae, a 2 × 2 × 2 factorial experiment was carried out with a climate treatment (future/his-torical; simulated as changes in both temperature and hydroperiod), indirect exposure to the invasive Brook Stickleback Culaea inconstans (Kirtland, 1840) fish (chemical cues present/absent), and conspecific density (conspecific present/ absent). We measured tadpole survival and development rate along with variables related to body shape, including mass, growth, growth rate, body mass index, tail fin depth, tail muscle depth, tail muscle width, and rear limb length. We as-sessed the effects of 3 common and important stressors (climate, invasive species, and density) alone and in combination with the other stressors to explore their interactions. Climate change, as simulated by our study, had large effects on Pacific Chorus Frog tadpole development and body shape. Brook Stickleback chemical cues had smaller but important effects on body shape and development rate when combined with additional stressors. Competition because of presence of a conspecific tadpole also reduced survival and had large effects on body shape. Pairwise interactions were largely additive or antagonistic. Three-way stressor interactions were most often antagonistic, resulting in less-than-additive effects of the 3 stressors together. However, 3 synergistic 3-way emergent interactions were detected. When all 3 stress-ors were present, development rate was faster than expected but rear limb length and tail muscle width were smaller than expected. Our results illustrate that climate change and conspecific density have wide-ranging effects on amphibian growth, development, and morphology—effects which may carry over into the terrestrial stage and reduce fitness. Furthermore, our findings reveal complex stressor interactions that were not predictable from single stressor effects or pairwise stressor effects, illustrating the difficulty in predicting multiple stressor interactions in freshwater systems.