DOI: 10.5061/dryad.7d7wm37zk
title: Phenotypic traits evolution and morphological traits associated with echolocation calls in cryptic horseshoe bats (Rhinolophidae)

description: Bats provide an excellent case study for studying evolution due to their
 remarkable flight and echolocation capabilities. In this study, we sought
 to understand the phenotypic evolution of key traits in Rhinolophidae
 (horseshoe bats) using phylogenetic comparative methods. We aim to test
 the phylogenetic signals of traits and evaluated the best-fit evolutionary
 models given the data for each trait considering different traits may
 evolve under different models (i.e., Brownian Motion (BM),
 Ornstein-Uhlenbeck (OU) and Early Burst (EB)) and reconstruct ancestral
 character states. We examined how phenotypic characters are associated
 with echolocation calls and minimum detectable prey size. We measured 34
 traits of 10 Asian rhinolophids species (187 individuals). We found that
 the majority of traits showed a high phylogenetic signal based on
 Blomberg’s K and Pagel’s λ, but each trait may evolve under different
 evolutionary models. Sella traits were shown to evolve under stabilizing
 selection based on OU models, indicating sella traits have the tendency to
 move forward along the branches toward some medial value in equilibrium.
 Our findings highlight the importance of sella characters in association
 with echolocation call emissions in Rhinolophidae, as calls are important
 for spatial cognition and also influence dietary preferences. Minimum
 detectable prey size in Rhinolophidae was associated with call frequency,
 bandwidth, call duration, wingspan and wing surface area. Ultimately,
 understanding trait evolution requires sensitivity due to the differential
 selective pressures which may apply to different characteristics.
description: Bats were captured using harp
 traps and released after measurements, photograph, tissue samples from
 biopsies punch and echolocation calls were recorded. The morphological
 measurements were made following Chornelia et al. (2022) which included external morphological
 measurements (forearm/FA, headbody/HB, hindfoot/HF, tibia/TB, ears/E,
 tail/Tail and mass/BM, noseleaf width/NLW, noseleaf height/NLL). External
 measurements were based on standard small mammal measurements following
 Francis (2019) using digital callipers Mitutoyo Absolute Series-500
 (accuracy 0.01 mm) and body mass was measured with Pesola Spring Scale
 (Pesola® Präzisionswaagen AG, Switzerland) with a precision of 0.3%. For
 noseleaf measurements, we used photographic metrics following (Chornelia
 et al., 2022) including 14 characters (Internarial cup width/INCW,
 noseleaf area/NLA, sella area/SA, sella height/Sh, noseleaf length/NLL,
 noseleaf width/NLW, lancet-base width/LBW, lancet height/LH, lancet
 tip-height/LTH, lancet angle/Lang, lancet tip-connecting process/Ltcp,
 ratio of LTH/LH, ratio of LBW/LH, and sella base width/SBW). From wing
 photographs (taken on gridded paper) a total of 11 wing parameters were
 included in the analyses including wingspan/B, wing tip ratio/I, wing
 aspect ratio/AR, wing loading/WL, arm-wing length/Law, hand-wing length/
 Lhw, arm-wing total surfaces area/Saw, hand-wing total surface area/Shw,
 ratio of Lhw and Law/TL, ratio of Shw and Saw/TS and total surface area of
 wing/S. Echolocation calls
 was collected from captured individual in the field and were recorded from
 hand-released. Calls were recorded using a Pettersson M500-384 (Pettersson
 Electronic AB, Uppsala, Sweden (www.batsound.com) and calls sequences were analysed using BatSound ver4 (Pettersson Electronic AB, Uppsala, Sweden) at sampling rate of 44.1 kHz and spectrogram was set at 1,024 sampling sites in Fast Fourier Transform (FFT) in conjunction with Hanning window. 
description: Microsoft Office, FigTree and any text editor.