Pre-copula acoustic behaviour of males in the malarial mosquitoes Anopheles coluzzii and Anopheles gambiae s.s. does not contribute to reproductive isolation.

We reveal that males of two members of the Anopheles gambiae s.l. species complex, Anopheles coluzzii and Anopheles gambiae s.s. (hereafter A. gambiae), which are both malaria vectors, perform a stereotypical acoustic behaviour in response to pure tones at frequencies that encompass the frequency range of the female's flight-tones. This behaviour resembles that described for Culex quinquefasciatus and consists of phonotactic flight initiated by a steep increase in wing-beat frequency (WBF) followed by rapid frequency modulation (RFM) of WBF when in close proximity to the sound source. RFM was elicited without acoustic feedback or the presence of a live female, but it appears to be a stereotypic behaviour in the immediate lead up to copula formation. RFM is an independent and different behavioural process from harmonic convergence interactions used by male-female pairs for mate recognition at earlier stages of mating. Acoustic threshold for RFM was used to plot behavioural audiograms from free-flying A coluzzii and A gambiae males. These audiograms were almost identical (minima ∼400 Hz) and encompassed the WBF ranges of A coluzzii (378-601 Hz) and A gambiae (373-590 Hz) females, indicating that males of the two species share similar frequency tuning and range. Furthermore, no differences were found between the two species in their WBFs, RFM behaviour or harmonic convergence ratios. These results indicate that assortative mating between A coluzzii and A gambiae is unlikely to be based on male-specific acoustic behaviours during RFM. The significance of these findings in relation to possible mechanisms for assortative mating is discussed.