The protocol described by Staunton et al., [28] was used to record the WBF of untethered virgin male and female 3–4 day old, Ae. aegypti from each strain. A single male or female mosquito was aspirated from a rearing cage into a vial (60 mL aspirator vial, Bioquip, 2809V, Compton, CA) to record ten seconds of uninterrupted flight (i.e., without landing and resting on the sides of the container). Flight sound was recorded using a TASCAM portable handheld recorder (DR- 22WL; Montebello, California). The sound file was recorded as a high resolution.wav file (32 bit), which was analysed using Audacity software version 2.3.2 ( The frequency spectrum of each recording was visualised, and the mean WBF determined from the individual peaks of the fundamental frequency. WBF was recorded for at least 30 male and 30 female individuals from each of the S-Cairns, R-BC and R-TL strains. All recordings were randomised and taken between 0600 and 1800 h.

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