According to the acoustic adaptation hypothesis, environmental and biogeographic factors such as atmospheric humidity can influence divergence of acoustic signals and speciation in high duty-cycle echolocating bats (e.g. Rhinolophus sp.), although this remains disputed. In this study we tested the hypothesis that Resting Frequency (RF) would decrease with increasing humidity along a large latitudinal gradient (6°-21° N), for four Rhinolophus species with different evolutionary histories, in the Western Ghats-Sri Lanka (WGSL) region.We conducted acoustic recordings and compiled published information on RFs of stationary Rhinolophus indorouxi, R. rouxi, R. beddomei, and R. lepidus from 40 roosts in 18 localities of the WGSL. These data comprised of recordings made with different devices and with different settings. Hence, due to the unknown measurement error involved in the recorded RFs, it was not possible to conduct conventional regression analyses to test our hypotheses.Hence, we qualitatively assessed effects of Relative Humidity (RH) and other environmental variables by interpreting only the sign, but not the magnitude of the RF responses (from the slopes of generalized least squares regression models). We also tested how RF and RH varied across biogeographic zones, and with bat body size. RFs of the Miocene-diverged species R.indorouxi and R. rouxi were higher at lower RH, as expected. In contrast, RF of the Pleistocene-diverged species R. beddomei and R. lepidus were higher at higher RH. Elevation and rainfall also emerged as important predictors of RF variation in these species. Bat body 2 size differed in dry and humid regions of the WGSL. RF variation was not consistent across biogeographic zones. The cryptic, phonically differentiated sibling species R. indorouxi and R. rouxi co-occurred only in mid-elevation zones along the Western Ghats escarpment. The variable but significant influences of humidity and correlated factors on RF suggest the importance of environmentally mediated acoustic divergence in different Rhinolophus species in the WGSL. We propose some hypotheses on interacting effects of environmental and phylogenetic factors on acoustic divergence in Rhinolophus bats of the WGSL. These ideas could be further tested with phylogenetic and acoustic studies, as more consistent and comparable data on these species become available in the future.