Seasonal variation in dengue virus transmission in northwestern Thailand is inversely related to the magnitude of diurnal temperature fluctuations, although mean temperature does not vary significantly across seasons. We tested the hypothesis that diurnal temperature fluctuations negatively influence epidemiologically important life-history traits of the primary dengue vector, Aedes aegypti (L.), compared with a constant 26°C temperature. A large diurnal temperature range (DTR) (≈18°C daily swing) extended immature development time (>1 d), lowered larval survival (≈6%), and reduced adult female reproductive output by 25% 14 d after blood feeding, relative to the constant 26°C temperature. A small DTR (≈8°C daily swing) led to a negligible or slightly positive effect on the life history traits tested. Our results indicate that there is a negative impact of large DTR on mosquito biology and are consistent with the hypothesis that, in at least some locations, large temperature fluctuations contribute to seasonal reduction in dengue virus transmission.