Design, construction and application of an automated thermal chamber to determine critical thermal maxima in small anurans

Critical thermal maximum (CTMax) is a thermal threshold useful for understanding physiological boundaries and predicting anuran vulnerability to climate warming. Given the value of this metric, a comprehensive review of the literature on CTMax protocols (1962-2025) revealed widespread methodological inconsistencies and rare verification of ramping rate accuracy, essential to gauge the reliability of estimates and facilitate comparisons among studies. We engineered an inexpensive (<300 USD) automated CTMax chamber incorporating dual thermocouples, a programmable proportional integral derivative (PID) controller, and real-time software monitoring to address these deficiencies. Performance validation across eight trials demonstrated alignment between programmed and observed temperature ramping rate (0.40 vs. 0.39 °C/min; Z = 1.58, P > 0.05). As a proof-of-concept, we used the proposed system to generate CTMax estimates of six species of Eleutherodactylus in Puerto Rico, including first reports for E. brittoni, E. cochranae, E. wightmanae, and E. juanariveroi. CTMax values ranged from 36.30°C ± 0.52 (E. wightmanae) to 42.10°C ± 0.66 (E. cochranae), with warming tolerances ranging from 15.00°C ± 0.63 to 18.18°C ± 0.73. Our design provides an accurate, replicable, and ethical approach for CTMax measurement and enhances the capacity to assess small amphibian vulnerability to global warming.