Mean daily temperatures predict the thermal limits of malaria transmission better than hourly rate summation.

Temperature shapes the geographic distribution, seasonality, and magnitude of mosquito-borne disease outbreaks. Models predicting transmission often use mosquito and pathogen thermal responses measured at constant temperatures. However, mosquitoes live in fluctuating temperatures. Rate summation--non-linear averaging of trait values measured at constant temperatures-is commonly used to infer performance in fluctuating environments, but its accuracy is rarely validated. We measured three traits that impact transmission-bite rate, survival, fecundity-in a malaria mosquito (Anopheles stephensi) across three diurnal temperature ranges (0, 9, and 12 °C). We compared transmission thermal suitability models with temperature-trait relationships observed under constant temperatures, fluctuating temperatures, and those predicted by rate summation. We mapped results across An. stephenesi's native Asian and invasive African ranges. We found: 1) daily temperature fluctuation trait values substantially differ from both constant temperature experiments and rate summation; 2) rate summation partially captured decreases in performance near thermal optima, yet incorrectly predicted increases near thermal limits; and 3) while thermal suitability across constant temperatures did not perfectly capture fluctuating environments, it was better than rate summation for estimating and mapping thermal limits. Our study provides insight into methods for predicting mosquito-borne disease risk and emphasizes the need to improve understanding of organismal performance under fluctuating conditions.