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Testicular temperature highly correlates with scrotal temperature. It has been postulated that cycling is associated with increased scrotal temperatures with time and consecutively with impaired semen quality. The aim of this study was to evaluate the influence of moderate cycling on scrotal temperature during highly standardized conditions in an experimental lab. A total of 25 volunteers without a history of infertility and normal andrological examination were included for scrotal temperature evaluation. Scrotal temperatures were measured every minute with a portable data recorder connected with two thermistor temperature sensors, which were attached on either side of the scrotum. A further thermistor sensor was attached on the central surface of the bicycle saddle. Ambient temperature in the study room was adjusted to 22 °C throughout the whole experiment. All volunteers started the experiment at the same daytime. Clothing of the volunteers consisted of standardized cotton wool trousers and shirts fitting to body size. After acclimatization to the study room in a sitting posture, each volunteer cycled on an exercise cycle for 60 min with a power of 25 Watt representing a speed of 25.45 km/h respectively. The saddle surface temperature reached in the median 35.59 °C after 60 min cycling. Median values of scrotal temperatures increased from 35.75 °C at the beginning to 35.82 °C after 60 min for the left side and from 35.50 to 35.59 °C for the right side. No correlation between cycling duration and scrotal temperatures could be found using multivariateanova for repeated measurements. However, scrotal temperatures during cycling were significantly lower ( p < 0.001) compared with the last 10 min in sitting posture before starting cycling with a difference of 1.31 °C for the left and 1.46 °C for the right side. The present study suggests that moderate cycling under standardized conditions with a power of 25 Watt is not a major genital heat stress factor. [ABSTRACT FROM AUTHOR]