Abstract Oxidative stress, driven by reactive oxygen species (ROS), poses a major challenge for organisms facing temperature fluctuations. This study provides the first direct in vivo measurements of ROS production in an insect, Chironomus riparius, across a broad range of ecologically relevant temperatures. We observed a U-shaped pattern of oxidative stress, with minimal ROS levels within an optimal thermal window (12-18C) and significantly elevated stress at both cold and warm extremes.