Normal sperm production depends on a testicular temperature below body temperature, but the thermogenic effects of daily life activities are not well known. We described the association between scrotal temperature and daily activities in 101 males using a non-invasive method for measuring scrotal temperature. A thermistor was attached to the underwear and the temperature of the scrotal skin was logged by a portable device every 5 min for 24h. Participants reported sedentary position and activities at work and during spare time in a questionnaire. Scrotal temperature was strongly correlated with sedentary work position with a dose-effect association (<1 h sedentary: 33.1 degrees C, >6 h sedentary: 34.7 degrees C, median values). The variation in sedentary work accounted for 31.5% of the variation in median temperature during the entire 24h. Sedentary position during spare time did not correlate with scrotal temperature. Median temperature at night was 1.2 degrees C higher than during the daytime. No effect was found for size or reported tightness of the underwear. In a model experiment, the deviance between testicular and scrotal temperature was estimated as maximally 0.1-0.6 degrees C, depending on the type of activity. Measuring scrotal temperature provides a valid estimate of testicular temperature and is feasible in large cohorts. We conclude that work position is an important determinant of testicular temperature.