We found a new acoustic signal in Drosophila simulans (si) and D. melanogaster (me). It is a ‘rejection signal’ (RS) produced by adult males and young males and females in response to the courting behaviour of mature males who emit ‘pulse songs’ (i.e. love song: LS). It occurs most frequently in si, less in adults me except if the interacting males belong to different chemical morphs (i.e. temperate or equatorial population). There are no differences in the LS characteristics directed to various sexes and ages. The RSs produced by adult males or by young animals do not differ significantly either. They are emitted by neither virgin nor fecundated adult me females but a few times by virgin adult si females. The RS (like the LS) is a multipulse signal but intervals between pulses are about twice those of LS, around 90 ms for si and 80 ms for me. They are very irregular, as is the distribution of energy along the bandwidth mainly between 300 and 800 Hz for si and 200 and 600 Hz for me. The sound level of the RS is from 10 to 20 dB less than the LS. The RS seems to be linked to the ‘flicking’ behaviour produced by both wings, while the LS always corresponds to the so-called ‘wing vibration’.