(Uploaded by Plazi for the Bat Literature Project) The acoustic behaviour of Eptesicus serotinus was investigated in the field using a 13.5-m vertical, linear microphone array that allowed for simultaneous recordings at three different heights and for the calculation of flight altitude and distance from the array. Recordings were made at two locations that differed in bat species diversity. E. serotinus hunted on average at an altitude of 10.7 m (±2.7) at one location and 6.8 m (±3.6) at the other location. Search signals were 5–17 ms long depending on flight altitude, and consisted of two to three frequency-modulated harmonics. For bats flying below 8–10 m altitude, signal duration decreased with decreasing flight altitude, whereas signal interval, terminal frequency, peak frequency and frequency range of the first harmonic increased. Above 8–10 m flight altitude, the signal parameters were fairly constant. The –10 dB bandwidth and duty cycle did not change with flight altitude. Source levels were calculated to between 121 and 125 dB peSPL re 20 µPa at 10 cm. For bats flying higher than 9 m, the microphone placed 1.5 m above the ground recorded significantly reduced signal durations and frequency ranges of the first harmonic compared to the same signals recorded with the microphones at heights of 7 or 15 m. We caution the use of ground recordings to fully describe the echolocation signals of high-flying bats. We demonstrate that flight altitude significantly influences the structure of sonar signals from E. serotinus.