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. 2022 Jan 12;8(2):eabl4844. doi: 10.1126/sciadv.abl4844

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Copyright © 2022 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).

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Fig. 4. — (A) The relative frequency distribution of harmonic convergence (HC) event numbers (N) from 513 virtual (“lone”) pairs (1-min flights) served as reference for significance analyses in real (“live”) pairs. To have a probability P < 0.05 of having occurred by chance, a live pair must exhibit at least nine convergence events. Values below nine are statistically unremarkable (P > 0.05). The average convergence event number (see figs. S14 and S15) for both lone and live pairs is ~3. Inset: Harmonic convergence event for a lone pair (M1, male fundamental; F1, female fundamental; M2, male second harmonic; F3, female third harmonic). (B) Violin plots of convergence event numbers in live pairs (top) and playback pairs (bottom) compared to convergence event numbers in lone pairs. No statistical differences are observed. Inset: Harmonic convergence event for a live (top) and a playback (bottom) pair. (C) For each mosquito pair, convergence events at a given ratio depend on the pair’s mean distance (d) from that ratio. Here, d is calculated as mean of the absolute distances of a pair’s instantaneous flight tone ratios (over the 1-min flight) from the given harmonic ratio (here, 3:2). The sharpness of the distribution peak highlights the extreme noise sensitivity of harmonic convergence calculations. Pairs whose flight tone ratios are accidentally close to the harmonic convergence ratio produce convergence events by mere chance. Pairs whose flight tones are accidentally far from the harmonic convergence ratio will not produce convergence events at all. (D) Left: Linear plot showing convergence event numbers for a real (live) pair of tethered-flying mosquitoes if the circularized time traces of their flight tones are phase-shifted against each other (phase shift, ΔΦ, 0° is the real-time pairing). Right: Polar plot for the same data. (E) Polar plots of two other real (live) pairs. Real-time pairing yields suboptimal convergence numbers for all pairs shown; maxima of harmonic convergence counts are randomly distributed across the phase space.