Interpreting empirical estimates of experimentally derived physiological and biological thermal limits in ectotherms

Whole-organism function is underpinned by physiological and biological processes, which respond to temperature over a range of time scales. Given that environmental temperature controls biological rates within ectotherms, different experimental protocols are needed to assess the ability of organisms to withstand extreme weather events versus gradual temperature change. Here we emphasize the importance of time in shaping ecological and evolutionary processes, and as an experimental parameter that is key when interpreting physiology studies reporting thermal limits. We discuss how acute and chronic thermal performance is underpinned by mechanisms operating at different time scales-resistance, acclimation, and adaptation. We offer definitions of common physiological and biological temperature metrics and identify challenges inherent to compiling the wealth of historical temperature limit data now available into meta-analytic frameworks. We use a case study, data across temperate fishes, to highlight that false positives may occur when differences in the thermal tolerances of species are in fact due to experimental protocols. We further illustrate that false negatives can arise if researchers fail to recognize differences in thermal limits of species emerging from macrophysiological approaches that are due to biological mechanisms. We strongly advocate for the careful design, interpretation, and reporting of experimental results to ensure that conclusions arising from data synthesis efforts are grounded in theory.