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The relative contributions of adaptation and genetic drift to morphological diversification of the skulls of echolocating mammals were investigated using two horseshoe bat species, Rhinolophus simulator and R. cf. simulator, as test cases. We used 3D geometric morphometrics to compare the shapes of skulls of the two lineages collected at various localities in southern Africa. Size and shape variation was predominantly attributed to selective forces; the between‐population variance (B) was not proportional to the within‐population variance (W). Modularity was evident in the crania of R. simulator but absent in the crania of R. cf. simulator and the mandibles of both species. The skulls of the two lineages thus appeared to be under different selection pressures, despite the overlap in their distributions. Difference in the crania of R. cf. simulator was centered largely on the nasal dome region of R. cf. simulator but on the cranium and mandibles of R. simulator. It is likely that the size and shape of the nasal dome, which acts as a frequency‐dependent acoustic horn, is more crucial in R. cf. simulator than in R. simulator because of the higher echolocation frequencies used by R. cf. simulator. A larger nasal dome in R. cf. simulator would allow the emission of higher intensity pulses, resulting in comparable detection distances to that of R. simulator. In contrast, selection pressure is probably more pronounced on the mandibles and cranium of R. simulator to compensate for the loss in bite force because of its elongated rostrum. The predominance of selection probably reflects the stringent association between environment and the optimal functioning of phenotypic characters associated with echolocation and feeding in bats., The relative contributions of adaptation and drift to morphological diversification of the crania of echolocating mammals were investigated using two horseshoe bat species, Rhinolophus simulator and R. cf. simulator, as test cases. Shape variation was predominantly attributed to selective forces. The predominance of selection probably reflects the stringent association between environment and the optimal functioning of phenotypic characters associated with echolocation and feeding in bats.

The performance of vocal motor control is crucial for vocal communication; however, its evolution and ecological drivers are largely unknown. Doppler shift compensation (DSC), which has been found only in bats, represents an excellent model for studying the mechanisms and evolution of vocal control. We tested whether species that are more dependent on DSC for pulse–echo separation have a more precise DSC and whether changes in environmental clutter would result in adjustments in DSC performance. We recorded the echolocation and flight behaviour of the greater horseshoe bat, Rhinolophus ferrumequinum, and the great leaf-nosed bat, Hipposideros armiger, flying through a corridor and a window in turn. The DSC precision in H. armiger was significantly lower than that of R. ferrumequinum; the offset between the reference frequency and resting frequency, however, was not significantly different between the two species. We also studied the DSC of H. armiger flying through windows of different sizes. Their flight speed decreased significantly with reduced window size. However, DSC precision and the offset did not vary significantly with window size. Our results suggest that DSC is more precise in rhinolophids, which have a higher demand for pulse–echo separation by DSC, than in hipposiderids. Changes in environmental clutter do not necessarily result in adjustments of DSC performance. Functional importance associated with high-performance vocal control may be critical in shaping the precision of vocal control in the course of evolution. Instead, habitat clutter encountered by vocalizing animals may not play an important role.

The order Chiroptera (bats) is the second largest group of mammals, composed of more than 1,300 species. Although powered flight and echolocation in bats have attracted many biologists, diversity in bat facial morphology has been almost neglected. Some bat species have a "nose leaf," a leaf-like epithelial appendage around their nostrils. The nose leaf appears to have been acquired at least three times independently in bat evolution, and its morphology is highly diverse among bats species. Internal tissue morphology of nose-leaves has been investigated through histological analyses of late-stage fetuses of some bat species possessing the nose leaf. However, the proximate factors that bring about chiropteran nose-leaves have not been identified. As an initial step to address the question above, we describe the normal embryonic development of the greater horseshoe bat Rhinolophus ferrumequinum, and examine development of the tissues associated with their nose leaf during embryogenesis through histological analyses. We found that the nose leaf of R. ferrumequinum is formed through two phases. First, the primordium of the nose leaf appears as two tissue bulges aligned top and bottom on the face at embryonic stages 15-16. Second, the sub-regions of the nose leaf are differentiated through ingrowth as well as outgrowth of the epithelium at stage 17. In embryogenesis of Carollia perspicillata, a phyllostomid species with a nose leaf, the nose leaf primordium is formed as a small tissue bulge on the nostril at stage 17. This tissue bulge grows into a dorsally projected thin epithelial structure. Such differences in the nose leaf developmental process between chiropteran lineages may suggest that distinct developmental mechanisms have been employed in each lineage's nose leaf evolution.

Horseshoe bats (Family Rhinolophidae) show an impressive array of morphological traits associated with use of high duty cycle echolocation calls that they emit via their nostrils (nasophonation). Delicate maxilloturbinal bones inside the nasal fossa of horseshoe bats have a unique elongated strand-like shape unknown in other mammals. Maxilloturbinal strands also vary considerably in length and cross-sectional shape. In other mammals, maxilloturbinals help direct respired air and prevent respiratory heat and water loss. We investigated whether strand-shaped maxilloturbinals in horseshoe bats perform a similar function to those of other mammals, or whether they were shaped for a role in nasophonation. Using histology, we studied the mucosa of the nasal fossa in Rhinolophus lepidus, which we compared with Hipposideros lankadiva (Hipposideridae) and Megaderma lyra (Megadermatidae). Using micro-CT scans of 30 horseshoe bat species, we quantified maxilloturbinal surface area and skull shape within a phylogenetic context. Histological results showed horseshoe bat maxilloturbinals are covered in a thin, poorly vascularized, sparsely ciliated mucosa poorly suited for preventing respiratory heat and water loss. Maxilloturbinal surface area was correlated with basicranial width, but exceptionally long and dorsoventrally flat maxilloturbinals did not show enhanced surface area for heat and moisture exchange. Skull shape variation appears to be driven by structures linked to nasophonation, including maxilloturbinals. Resting echolocation call frequency better predicted skull shape than did skull size, and was specifically correlated with dimensions of the rostral inflations, palate, and maxilloturbinals. These traits appear to form a morphological complex, indicating a nasophonatory role for the strand-shaped rhinolophid maxilloturbinals. Anat Rec, 2018. © 2018 American Association for Anatomy.

The relative contributions of adaptation and drift to morphological diversification of the crania of echolocating mammals was investigated using two horseshoe bat species, Rhinolophus simulator and R. cf. simulator as test cases. We used 3D geometric morphometrics to compare the shapes of skulls of the two lineages collected at various localities in southern Africa. Shape variation was predominantly attributed to selective forces; the between population variance (B) was not proportional to the within population variance (W). Modularity was evident in the crania of R. simulator but absent in the crania of R. cf. simulator and the mandibles of both species. The skulls of the two lineages thus appeared to be under different selection pressures, despite the overlap in their distributions. Selection acted mainly on the nasal dome region of R. cf. simulator whereas selection acted more on the cranium and mandibles than on the nasal domes of R. simulator. Probably the relatively higher echolocation frequencies used by R. cf. simulator, the shape of the nasal dome, which acts as a frequency dependent acoustic horn, is more crucial than in R. simulator, allowing maximization of the intensity of the emitted calls and resulting in comparable detection distances. In contrast, selection pressure is probably more pronounced on the mandibles and cranium of R. simulator to compensate for the loss in bite force because of its elongated rostrum. The predominance of selection probably reflects the stringent association between environment and the optimal functioning of phenotypic characters associated with echolocation and feeding in bats.

Recently diverged taxa with contrasting phenotypes offer opportunities for unravelling the genetic basis of phenotypic variation in nature. Horseshoe bats are a speciose group that exhibit a derived form of high-duty cycle echolocation in which the inner ear is finely tuned to echoes of the narrowband call frequency. Here, by focusing on three recently diverged subspecies of the intermediate horseshoe bat (Rhinolophus affinis) that display divergent echolocation call frequencies, we aim to identify candidate loci putatively involved in hearing frequency variation. We used de novo transcriptome sequencing of two mainland taxa (himalayanusandmacrurus) and one island taxon (hainanus) to compare expression profiles of thousands of genes. By comparing taxa with divergent call frequencies (around 15 kHz difference), we identified 252 differentially expressed genes, of which six have been shown to be involved in hearing or deafness in human/mouse. To obtain further validation of these results, we applied quantitative reverse transcription–PCR to the candidate geneFBXL15and found a broad association between the level of expression and call frequency across taxa. The genes identified here represent strong candidate loci associated with hearing frequency variation in bats.

Cochlear morphology has been regarded as one of the key traits to understand the origin and evolution of echolocation in bats, given its functionality and performance for receiving echolocation sonar. While numerous researchers have compared adult-stage morphology, few have studied the prenatal development of the cochlea. Here, we provide the first detailed three-dimensional description of the prenatal cranial development in bats, using Rhinolophus thomasi as a model, with particular interest to the petrosal which houses the cochlea. Results revealed that among all cranial bones the onset of the ossification of the petrosal is earlier in R. thomasi when compared to other reported mammals. Generally, the cochlea reaches adult size and shape before or around birth in placental mammals including bats, but we found that its shape and size growths continue until maturity in Rhinolophus species. The relationship of cochlear size and skull size is maintained constant throughout the postnatal ontogeny to adulthood in Rhinolophus, a pattern previously reported neither in any other bats nor other mammals. The peculiar developmental pattern in Rhinolophus possibly allows them to form their characteristically large cochlea and facilitate their distinctive echolocation behavior. A recent study reported that non-echolocating Pteropodidae shares a similar prenatal cochlear size to laryngeal echolocating bats. The apparent resemblance of fetal cochlear size was proposed to be a vestigial signal of large cochlear size in the last common ancestor of bats and thus as supporting evidence for the single origin of laryngeal echolocation. However, results from the present observations suggest that limited aspects of the cochlear development were captured in this previous investigation and that the resulting interpretations may be questionable. We point out that diversity and patterns of cochlear development among bats are still not resolved, and the controversy on the origins of laryngeal echolocation is still open to discussion.

Drift, selection, or their combined effects commonly drive geographic variation in traits. Clarifying the relative roles of each process is a long-standing research goal in evolutionary biology. Acoustic signals of bats are a phenotypic characteristic that plays an important role in social organization and species recognition. We extensively sampled the Chinese rufous horseshoe bat (Rhinolophus sinicus) throughout China and Vietnam and reconstructed a species phylogeny to better understand the patterns and causes of the geographic variation of acoustic signals. Our results showed that the resting frequency (RF) of calls varied with latitude, sex, and distance among different colony locations. RF differences were not correlated with genetic distance (based on only one nuclear locus and the mitochondrial locus), climatic factors (mean annual temperature and mean annual precipitable water), or body size, although differences in calls increased with distance among various populations. This suggests that cultural drift may play more important roles than genetic drift and acoustic adaptation in shaping acoustic differences within regions in R. sinicus.

Background Although the sensory drive hypothesis can explain the geographic variation in echolocation frequencies of some bat species, the molecular mechanisms underlying this phenomenon are still unclear. The three lineages of greater horseshoe bat (Rhinolophus ferrumequinum) in China (northeast, central-east, and southwest) have significant geographic variation in resting frequencies (RF) of echolocation calls. Because their cochleae have an acoustic fovea that is highly sensitive to a narrow range of frequencies, we reported the transcriptomes of cochleae collected from three genetic lineages of R. ferrumequinum, which is an ideal organism for studying geographic variation in echolocation signals, and tried to understand the mechanisms behind this bat phenomenon by analyzing gene expression and sequence variation. Results A total of 8190 differentially expressed genes (DEGs) were identified. We identified five modules from all DEGs that were significantly related to RF or forearm length (FL). DEGs in the RF-related modules were significantly enriched in the gene categories involved in neural activity, learning, and response to sound. DEGs in the FL-related modules were significantly enriched in the pathways related to muscle and actin functions. Using 21,945 single nucleotide polymorphisms, we identified 18 candidate unigenes associated with hearing, five of which were differentially expressed among the three populations. Additionally, the gene ERBB4, which regulates diverse cellular processes in the inner ear such as cell proliferation and differentiation, was in the largest module. We also found 49 unigenes that were under positive selection from 4105 one-to-one orthologous gene pairs between the three R. ferrumequinum lineages and three other Chiroptera species. Conclusions The variability of gene expression and sequence divergence at the molecular level might provide evidence that can help elucidate the genetic basis of geographic variation in echolocation signals of greater horseshoe bats.

The diversity of bats worldwide includes large numbers of cryptic species, partly because divergence in acoustic traits such as echolocation calls are under stronger selection than differences in visual appearance in these nocturnal mammals. Island faunas often contain disproportionate numbers of endemic species, and hence we might expect cryptic, endemic species to be discovered relatively frequently in bats inhabiting islands. Species are best defined when multiple lines of evidence supports their diagnosis. Here we use morphometric, acoustic, and molecular phylogenetic data to show that a horseshoe bat in the Andaman Islands is distinct in all three aspects, supporting its description as a newly described endemic species. We recommend investigation into possible new and endemic bat species on islands by using integrated approaches that provide independent lines of evidence for taxonomic distinctiveness. We provide a formal description of the new species –Rhinolophus andamanensisDobson, 1872.

The geographical variation in animals’ social acoustical signals can reduce recognition across populations and may thus promote assortative mating, reproductive isolation and speciation. However, the social consequences of geographical variation in acoustical signals serving an ecological function are poorly known. Bat echolocation calls are considered to have a dual function; they are used not only for orientation and foraging but also for communication. In this study, we studied the behavioural response of Rhinolophus ferrumequinum to geographical variation in echolocation calls. Using habituation-dishabituation playback experiments, we found that all tested bats from northeast China exhibited obvious responses after switching playback from their own population to those from central east China and southwest China. Using two-choice playback experiments, we showed that the bats from northeast China responded more strongly to echolocation calls from their own population than to those from central east China. Our results demonstrate that Rhinolophus ferrumequinum is able to discriminate between echolocation calls of its local population and a foreign one and that local echolocation calls are preferred over foreign ones in a two-choice context. This study supports the communicative potential of bat echolocation calls and provides insight into the discrimination ability and behavioural preference of bats with respect to geographical variation in echolocation calls. This study provides behavioural evidence for the communicative role in echolocation calls and indicates that a dual function of echolocation calls exists more commonly in bats. Moreover, this study provides insight into the discrimination ability and preference of bats with respect to frequency variation in echolocation calls and contributes to a better understanding of how geographical evolution in ecological acoustical signals may affect the ability of recognition between populations.

The noseleaf and pinnae of horseshoe bats (Rhinolophus ferrumequinum) have both been shown to actively deform during biosonar operation. Since these baffle structures directly affect the properties of the animals biosonar system, this work mimics horseshoe bat sonar system with the goal of developing a platform to study the dynamic sensing principles horseshoe bats employ. Consequently, two robotic devices were developed to mimic the dynamic emission and reception characteristics of horseshoe bats. The noseleaf and pinnae shapes were modeled as smooth blanks matched to digital representations of a horseshoe bat specimens noseleaf and pinnae. Local shape features mimicking structures on the pinnae and noseleaf were added digitally. Flexible baffles with local shape feature combinations were manufactured and paired with actuation mechanisms to mimic pinnae and noseleaf deformations in vivo. Two noseleaves with and without local shape features were considered. Each noseleaf baffle was mounted to a platform called the dynamic emission head to actuate three surface elements of the baffle. Similarly, 12 pinna realizations composed of combinations of three local shape features were mounted to a platform called the dynamic reception head to deform the left and right pinnae independently. Motion of the noseleaf and pinnae were synchronized to the incoming and outgoing sonar waveform, and the joint time-frequency properties of the noseleaf and pinnae local feature combinations and pairs of pinnae and noseleaf thereof were characterized across spatial direction. Amplitude modulations to the outgoing and incoming sonar pulse information across spatial direction were observed for all pinnae and noseleaf local shape feature combinations. Peak modulation variance generated by motion of the pinnae and combinations of the noseleaf and pinnae approached a white Gaussian noise variance bound. It was found the dynamic emitter generated less modulation than either the combined or reception scenarios.

Geographic variation in sensory traits is usually influenced by adaptive processes because these traits are involved in crucial life-history aspects including orientation, communication, lineage recognition and mate choice. Studying this variation can therefore provide insights into lineage diversification. According to the Sensory Drive Hypothesis, lineage diversification may be driven by adaptation of sensory systems to local environments. It predicts that acoustic signals vary in association with local climatic conditions so that atmospheric attenuation is minimized and transmission of the signals maximized. To test this prediction, we investigated the influence of climatic factors (specifically relative humidity and temperature) on geographic variation in the resting frequencies of the echolocation pulses of Geoffroy's horseshoe bat, Rhinolophus clivosus. If the evolution of phenotypic variation in this lineage tracks climate variation, human induced climate change may lead to decreases in detection volumes and a reduction in foraging efficiency. A complex non-linear interaction between relative humidity and temperature affects atmospheric attenuation of sound and principal components composed of these correlated variables were, therefore, used in a linear mixed effects model to assess their contribution to observed variation in resting frequencies. A principal component composed predominantly of mean annual temperature (factor loading of -0.8455) significantly explained a proportion of the variation in resting frequency across sites (P < 0.05). Specifically, at higher relative humidity (around 60%) prevalent across the distribution of R. clivosus, increasing temperature had a strong negative effect on resting frequency. Climatic factors thus strongly influence acoustic signal divergence in this lineage, supporting the prediction of the Sensory Drive Hypothesis. The predicted future increase in temperature due to climate change is likely to decrease the detection volume in echolocating bats and adversely impact their foraging efficiency.

Communication is a fundamental component of evolutionary change because of its role in mate choice and sexual selection. Acoustic signals are a vital element of animal communication and sympatric species may use private frequency bands to facilitate intraspecific communication and identification of conspecifics (acoustic communication hypothesis, ACH). If so, animals should show increasing rates of misclassification with increasing overlap in frequency between their own calls and those used by sympatric heterospecifics. We tested this on the echolocation of the horseshoe bat, Rhinolophus capensis, using a classical habituation–dishabituation experiment in which we exposed R. capensis from two phonetic populations to echolocation calls of sympatric and allopatric horseshoe bat species (Rhinolophus clivosus and Rhinolophus damarensis) and different phonetic populations of R. capensis. As predicted by the ACH, R. capensis from both test populations were able to discriminate between their own calls and calls of the respective sympatric horseshoe bat species. However, only bats from one test population were able to discriminate between calls of allopatric heterospecifics and their own population when both were using the same frequency. The local acoustic signalling assemblages (ensemble of signals from sympatric conspecifics and heterospecifics) of the two populations differed in complexity as a result of contact with other phonetic populations and sympatric heterospecifics. We therefore propose that a hierarchy of discrimination ability has evolved within the same species. Frequency alone may be sufficient to assess species membership in relatively simple acoustic assemblages but the ability to use additional acoustic cues may have evolved in more complex acoustic assemblages to circumvent misidentifications as a result of the use of overlapping signals. When the acoustic signal design is under strong constraints as a result of dual functions and the available acoustic space is limited because of co-occurring species, species discrimination is mediated through improved sensory acuity in the receiver.

Divergence in phenotypic traits may arise from the interaction of different evolutionary forces, including different kinds of selection (e.g., ecological), genetic drift, and phenotypic plasticity. Sensory systems play an important role in survival and reproduction, and divergent selection on such systems may result in lineage diversification. Such diversification could be largely influenced by selection in different environments as a result of isolation by environment (IbE). We investigated this process using geographic variation in the resting echolocation frequency of the horseshoe bat species, Rhinolophus damarensis, as a test case. Bats were sampled along a latitudinal gradient ranging from 16°S to 32°S in the arid western half of southern Africa. We measured body size and peak resting frequencies (RF) from handheld individual bats. Three hypotheses for the divergence in RF were tested: (1) James’ Rule, (2) IbE, and (3) genetic drift through isolation by distance (IbD) to isolate the effects of body size, local climatic conditions, and geographic distance, respectively, on the resting frequency of R. damarensis. Our results did not support genetic drift because there was no correlation between RF variation and geographic distance. Our results also did not support James' Rule because there was no significant relationship between (1) geographic distances and RF, (2) body size and RF, or (3) body size and climatic variables. Instead, we found support for IbE in the form of a correlation between RF and both region and annual mean temperature, suggesting that RF variation may be the result of environmental discontinuities. The environmental discontinuities coincided with previously reported genetic divergence. Climatic gradients in conjunction with environmental discontinuities could lead to local adaptation in sensory signals and directed dispersal such that gene flow is restricted, allowing lineages to diverge. However, our study cannot exclude the role of processes like phenotypic plasticity in phenotypic variation.

The auditory system of the Korean greater horseshoe bat (Rhinolophus ferrumequinum korai, RFK) is adapted to its own echolocation signal, which consist of constant frequency (CF) element and frequency modulated (FM) element. In contrast, the Japanese long-fingered bat (Miniopterus schreibersii fuliginosus, MSF) emits FM signals. In the present study, the characteristics of stereocilia in RFK (a CF/FM bat) and MSF (a FM bat) were studied in the apical turn of the cochlea where the lower frequencies are transduced. Stereocilia lengths and numbers were quantitatively measured in RFK by scanning electron microscopy and compared with those of MSF. Each inner hair cells (IHCs) of RFK possessed three rows of stereocilia, whereas MSF possessed five rows of stereocilia. Gradients in stereocilia lengths and numbers of stereocilia of the IHCs of RFK were found to be less pronounced and fewer, respectively, than those of MSF. Each outer hair cells (OHCs) possessed three rows of stereocilia in both species. OHCs stereocilia in RFK that distinguished it from MSF were a shorter length and a greater number of stereocilia. These features suggest that the apical cochleas of RFK are adapted for the processing of higher frequency echolocation calls rather than that of MSF.

Mutual recognition is the product of species coexistence, and has direct effects on survival and reproduction of animals. Bats are able to discriminate between sympatric different heterospecifics based on their echolocation calls, which has been shown both in free-flying and captive bats. To date, however, the factors that may determine the behavioral responses of bats to echolocation calls from sympatric heterospecifics have rarely been tested, especially under well-controlled conditions in captive bats. Hence, we aimed at tackling this question by performing playback experiments (habituation–dishabituation) with three horseshoe bat species within the constant-frequency bat guild, which included big-eared horseshoe bats (Rhinolophus macrotis), Blyth’s horseshoe bats (Rhinolophus lepidus), and Chinese horseshoe bats (Rhinolophus sinicus). We studied the behavioral responses of these three species to echolocation calls of conspecifics, to other two species, and to another heterospecifics bat, Stoliczka’s trident bat (Asellisus stoliczkanus), which also belongs to this guild. We found that the three rhinolophid species displayed a series of distinct behaviors to heterospecific echolocation but few to conspecific calls after habituation, suggesting that they may have been able to discriminate sympatric heterospecific echolocation calls from those of conspecifics. Interestingly, the behavioral responses to heterospecific calls were positively correlated with the interspecific overlap index in trophic niche, whereas call design had only a minor effect. This implies that the behavioral responses of these bats to heterospecific echolocation calls may be related to the degree of interspecific food competition.

Acoustic signals are important to the biology of animals, mediating crucial activities such as social interactions (communication) as well as orientation and foraging (echolocation). Many signals used in communication are vocal, which are especially important in nocturnal animals such as bats where social interactions occur in darkness. Despite this, little is known about the social calls and behaviours of echolocating bats. To better understand their social and acoustic behaviour, we compiled an ethogram and list of vocalisations for Geoffroy's horseshoe bat (Rhinolophus clivosus). We kept three non-contemporaneous and short-term captive groups to capture interactions and social calls using simultaneous video and audio recordings. The resultant ethogram was comprised of 40 unique types of behaviour, both social and non-social. Social calls (n = 255) were assigned to different call types by their frequency-time contour and categorised by their behaviours, situational categories and functional contexts (affiliative or agonistic). From the calls observed, four acoustically distinct call types were identified: (i) cascading/rising frequency-modulated (FM) calls (n = 26), (ii) oscillatory FM calls (n = 140), (iii) noisy screech calls (n = 68), and (iv) whistle calls (n = 21) (GLM: F30, 711 = 24.28, P < 0.001). Call types showed only weak associations with certain behaviours, situational categories or functional contexts. However, calls with specific acoustic attributes accompanied the behaviour of wing swat [discriminant function analysis (DFA) classification success: 76%; GLM: F20, 108 = 4.12, P < 0.001] and situational category of flight (DFA classification success: 82%; GLM: F20, 136 = 2.97, P < 0.001). An analysis of acoustic attributes across all call types showed weak associations with functional context (Affiliative DFA classification success: 6%). Only one acoustic parameter (peak frequency) had a slight significant difference between calls emitted during agonistic and affiliative interactions (GLM: F10, 73 = 2.30, P < 0.05; post-hoc unequal n: P = 0.044). In addition to the description of distinct call types, we provide evidence of transitional calls in this bat species in which a social call transitions seamlessly into an echolocation pulse. This study gives a first glimpse into the behaviours and social vocalisations produced by R. clivosus. Basic behavioural data such as these may facilitate the design of experiments that allow greater insight into the social organisation of bats.

Animal communication follows many coding schemes. Less is known about the coding strategy for signal length and rates of use in animal vocal communication. A generalized brevity (negative relation between signal length and frequency of use) is innovatively explored but remains controversial in animal vocal communication. We tested brevity for short-range social and distress sounds from four echolocating bats: adult black-bearded tomb bat Taphozous melanopogon, Mexican free-tailed bat Tadarida brasiliensis, adult greater horseshoe bat Rhinolophus ferrumequinum, and adult least horseshoe bat Rhinolophus pusillus. There was a negative association between duration and number of social but not distress calls emitted. The most frequently emitted social calls were brief, while most distress calls were long. Brevity or lengthiness was consistently selected in vocal communications for each species. Echolocating bats seem to have convergent coding strategy for communication calls. The results provide the evidence of efficient coding in bat social vocalizations, and lay the basis of future researches on the convergence for neural control on bats' communication calls.

Geographic variation can be an indicator of still poorly understood evolutionary processes such as adaptation and drift. Sensory systems used in communication play a key role in mate choice and species recognition. Habitat-mediated (i.e. adaptive) differences in communication signals may therefore lead to diversification. We investigated geographic variation in echolocation calls of African horseshoe bats, Rhinolophus simulator and R . swinnyi in the context of two adaptive hypotheses: 1) James' Rule and 2) the Sensory Drive Hypothesis. According to James' Rule body-size should vary in response to relative humidity and temperature so that divergence in call frequency may therefore be the result of climate-mediated variation in body size because of the correlation between body size and call frequency. The Sensory Drive Hypothesis proposes that call frequency is a response to climate-induced differences in atmospheric attenuation and predicts that increases in atmospheric attenuation selects for calls of lower frequency. We measured the morphology and resting call frequency (RF) of 111 R . simulator and 126 R . swinnyi individuals across their distributional range to test the above hypotheses. Contrary to the prediction of James' Rule, divergence in body size could not explain the variation in RF. Instead, acoustic divergence in RF was best predicted by latitude, geography and climate-induced differences in atmospheric attenuation, as predicted by the Sensory Drive Hypothesis. Although variation in RF was strongly influenced by temperature and humidity, other climatic variables (associated with latitude and altitude) as well as drift (as suggested by a positive correlation between call variation and geographic distance, especially in R . simulator ) may also play an important role.

Horseshoe bats (Rhinolophidae), Old World leaf-nosed bats (Hipposideridae), and several species of moustached bats (Mormoopidae), including Parnell’s moustached bat (Pteronotus parnellii), Paraguayan moustached bat (Pteronotus paraguayensis) and Mesoamerican moustached bats (Pteronotus mesoamericanus) emit pulses with long constant frequency (CF) components and have evolved mechanisms for separating the pulse and echo in frequency. This approach to echolocation—high duty cycle (HDC) echolocation—depends largely on Doppler shift compensation (DSC). In 1968, Schnitzler discovered that the greater horseshoe bat (Rhinolophus ferrumequinum) compensates for flight-induced Doppler shifts in the CF component of echoes by adjusting their call frequency, ensuring a stable echo frequency during flight. This significant behavioral adaptation is supported by an acoustic fovea, a striking morphological and physiological specialization occurring from the peripheral to the central auditory system in HDC bats. The auditory fovea has neurons with high sensitivity to the narrow frequency range of the CF components of echolocation calls of HDC bats. Doppler shift compensation of the frequency that dominates the echolocation calls maintains the frequency within the narrow range of the acoustic fovea. This combination of CF calls, HDC echolocation, and DSC allows fine-frequency analysis of acoustic glints in echoes from insects fluttering their wings, which makes these bats very effective at detecting flutter.

Echolocation calls produced by bats contain information on the emitter that could be used in social interactions. Nevertheless, little is known about the whether such information actually is being used by bats. Here we tested the ability of two horseshoe bat species (Rhinolophus mehelyi and R. euryale; n = 20 each) to discriminate conspecific male from conspecific female echolocation calls and vice versa. Behavioural data from a habituation-discrimination paradigm revealed that both species were able to recognize the sex of conspecifics from their calls, providing evidence that cues on the sex of the emitter are encoded in echolocation signals. While our data support that the sex-specific acoustic cues are not (just) the horseshoe bats' constant-frequency call part, their nature remains enigmatic. This is the second study to show that bats can recognize the caller's gender from echolocation calls and the first to do so for ‘high-duty-cycle bats’; those that use calls with a prominent constant-frequency (C...

Horseshoe bats emit biosonar pulses through the nostrils and diffract the outgoing ultrasonic pulses with baffles, so-called “noseleaves,” that surround the nostrils. The noseleaves have complex static geometries and can furthermore undergo dynamic shape changes during emission of the biosonar pulses. The posterior noseleaf part, the lancet, has been shown to carry out anterior-posterior flicking motions during biosonar emissions with average lancet tip displacements of about 1 mm. Here, the acoustic effects of the interplay between the lancet furrows and shape change (lancet rotation) on the emission beam were investigated using the animated digital models obtained from the noseleaves of greater horseshoe bats (Rhinolophus ferrumequinum). It was found that forward lancet rotations increase the amount of sound energy allocated to secondary amplitude maxima (sidelobes) in the beampattern, but only in the presence of the furrows. The interaction between static and dynamic features can be readily quantified by roughness (standard deviation about local mean) of the amplitude distribution of the beampatterns. This effect goes beyond the static impact of the furrows on the width of the mainlobe. It could allow the bats to send out their pulses through a sequence of qualitatively different beampatterns.

In this study, we analyzed the pulse-duration, pulse-interval and peak-frequency of echolocation call in three species as Rhinolophus ferrumequinum, Pipistrellus abramus, and Myotis macrodactylus. The peak frequency and pulse duration for above mentioned species were 69 kHz, 47 kHz and 49 kHz and , and for R. ferrumequinum, P. abramus and M. macrodactylus, respectively. The pulse intervals for R. ferrumequinum, P. abramus and M. macrodactylus were , and , respectively. The pulse pattern of R. ferrumequinum was setting into a short FM call and linked to long CF call and went through the short FM call again. The pulse pattern of M. macrodactylus was comprised with serial short FM call and the CF call was not checked up in accordance with the spectrogram analysis. The long FM call and short CF call got join together for the P. abramus and the peak frequency was checked up at the pulse ending as CF call.

The origin and meaning of echolocation call frequency variation within rhinolophid bats is not well understood despite an increasing number of allopatric and sympatric examples being documented. A bimodal distribution of mean regional call frequency within the Okinawa-jima Island population of Rhinolophus cornutus pumilus (Rhinolophidae) provided a unique opportunity to investigate geographic call frequency variation early in its development. Individual resting echolocation frequencies, partial mitochondrial DNA D-loop sequences and genotypes from six microsatellite loci were obtained from 288 individuals in 11 colonies across the entire length of the island, and nearby Kume-jima Island. Acoustic differences (5-8 kHz) observed between the north and south regions have been maintained despite evidence of sufficient nuclear gene flow across the middle of the island. Significant subdivision of maternally inherited D-loop haplotypes suggested a limitation of movement of females between regions, but not within the regions, and was evidence of female philopatry. These results support a 'maternal transmission' hypothesis whereby the difference in the constant frequency (CF) component between the regions is maintained by mother-offspring transmission of CF, the restricted dispersal of females between regions and small effective population size. We suggest that the mean 5-8 kHz call frequency difference between the regions might develop through random cultural drift.

A cryptic species of the big-eared horseshoe bat (Rhinolophus macrotis) was identified in Jiangxi Province, China, based on significant differences in echolocation frequencies and morphology. Consistent with the bimodal distribution of body sizes of R. macrotis specimens obtained from the same cave, we now consider this population to be comprised of two putative species; a large and a small form, with dominant echolocation call frequencies of 49 kHz and 65 kHz, respectively. Cytochrome b sequences of these two phonic forms diverged by 3.16–3.25%, a similar level of divergence to that between the large form and the outgroup, R. rex (3.33–3.77%). These differences strongly suggest that the two phonic forms are distinct species. We also found that the wing loading and aspect ratio of the small form was much lower than that of the large form, suggesting that the small form is capable of foraging in denser forest. Without dietary evidence, the ecological significance of the observed difference in echo...

Bat-and-moth is a good model system for understanding predator-prey interactions resulting from interspecific coevolution. Night-flying insects have been under predation pressure from echolocating bats for 65Myr, pressuring vulnerable moths to evolve ultrasound detection and evasive maneuvers as counter tactics. Past studies of defensive behaviors against attacking bats have been biased toward noctuoid moth responses to short duration pulses of low-duty-cycle (LDC) bat calls. Depending on the region, however, moths have been exposed to predation pressure from high-duty-cycle (HDC) bats as well. Here, we reveal that long duration pulse of the sympatric HDC bat (e.g., greater horseshoe bat) is easily detected by the auditory nerve of Japanese crambid moths (yellow peach moth and Asian corn borer) and suppress both mate-finding flights of virgin males and host-finding flights of mated females. The hearing sensitivities for the duration of pulse stimuli significantly dropped non-linearly in both the two moth species as the pulse duration shortened. These hearing properties support the energy integrator model; however, the threshold reduction per doubling the duration has slightly larger than those of other moth species hitherto reported. And also, Asian corn borer showed a lower auditory sensitivity and a lower flight suppression to short duration pulse than yellow peach moth did. Therefore, flight disruption of moth might be more frequently achieved by the pulse structure of HDC calls. The combination of long pulses and inter-pulse intervals, which moths can readily continue detecting, will be useful for repelling moth pests.

Most regions of the world support moth species that use caves as diurnal roosts and/or as overwintering and mating sites. Although subterranean roosts are often also occupied by insectivorous bats, nothing is known about moth–bat interaction at these sites. The frequency of cohabitation and the dynamics of moth–bat interaction at roosts were examined during surveys at 30 known bat roosts in eastern Australia; 15 disused mines and 15 natural caves. Moths and bats cohabited in 20 roosts (67%); however, only two species of eared moths, Speiredonia spectans and S. mutabilis (Noctuidae), were observed. These large (wingspan up to 75 mm) species show a classic escape response to ultrasound. Moths used roosts occupied by bats with both frequency modulated (FM) (four species) and constant frequency (CF) (two species) echolocation calls. Signal frequencies of these bats had ranges of 23–25 and 44–71 kHz. Moths were absent from roosts occupied by high-frequency (> 140 kHz) CF bats. Predator–prey interaction was assessed by studying a sub-sample of six roosts occupied by S. spectans and the horseshoe bat Rhinolophus megaphyllus. Speiredonia spectans and R. megaphyllus cohabited roosts throughout the year. Bats captured and consumed moths in these roosts. The numbers of moth wings on the floor of roosts were counted to assess rates of predation. Bats consumed 209 moths at the six roosts over a 12-month period, representing 19% of the moth population at the start of the study. The study establishes that moth–bat cohabitation of roosts is relatively common in eastern Australia and that bats do capture moths in these sites. Bat predation seems to be a significant source of mortality of moths that occupy subterranean roosts.

Factors influencing prey selection by insectivorous bats are poorly understood but may be related to interspecific differences in echolocation call structure. The diet of Eptesicus fuscus was compared with prey abundance across a foraging season to examine prey selection by a bat species which has an echolocation call dominated by a frequency-modulated (FM) component. Guano samples were collected on 18 occasions from a single colony between May and September 2000. Abundance of aerial insects was measured using a black-light trap on nights when guano was collected. Regression analysis was used to test the hypotheses that E. fuscus: (1) does not take prey in proportion to their relative abundance; (2) forages preferentially for beetles in a manner predicted by foraging theory. Relative abundance (= proportion in relation to other prey types) of prey in trap samples was not a good predictor of prey in the diet across periods or within periods, which suggests selective foraging. Beetles (42–96% of the diet by per cent volume) were consistently eaten in higher proportions than their relative abundance. The absolute abundance of beetles (= abundance irrespective of the abundance of other prey types) explained 60% of the variation in beetle use with more beetles eaten when they were abundant. Absolute beetle abundance explained 66%, 53% and 47% of the variation in the use of three other prey types, with more of these prey eaten when beetle abundance was low. Finally, absolute beetle abundance explained 60% of the variation in diet breadth, which increased when beetle abundance was low. Our results agree with predictions derived from foraging theory and support the hypothesis that E. fuscus forages selectively for beetles, consuming a wider variety of prey when beetle abundance is low. These results are similar to those found for the greater horseshoe bat Rhinolophus ferrumequinum which has an echolocation call dominated by a constant-frequency (CF) component, and suggest that some FM bats may discriminate among prey taxa, at least at the ordinal level.

Bats have evolved unique mechanisms by which to navigate and hunt in their environments. Some species of the microchiroptera, such as members of the horseshoe bat family (Rhinolophidae), which live in particularly challenging, cluttered environments, have been observed to move their peripheral structures for emission and reception (noseleaf and pinna) during echolocation very rapidly, e.g., within a tenth of a second. With these behaviors, the bats could create time-variant properties in their biosonar systems to provide additional degrees of freedom to be used for enhancing the encoding of sensory information. To make full use of this opportunity, a tight sensorimotor integration is likely to be required. In order to explore the possible role of sensorimotor integration in a dynamic biosonar system, we have developed biomimetic robotic models to replicate the effects of the dynamics of the baffle structures. Data obtained with these systems indicates that a coordinated emitter and receiver dynamics can b...

One of the most efficient mechanisms to optimize signal-to-noise ratios is the Lombard effect - an involuntary rise in call amplitude due to ambient noise. It is often accompanied by changes in the spectro-temporal composition of calls. We examined the effects of broadband-filtered noise on the spectro-temporal composition of horseshoe bat echolocation calls, which consist of a constant-frequency component and an initial and terminal frequency-modulated portion. We find that the frequency-modulated components became larger for almost all noise conditions, whereas the bandwidth of the constant-frequency component increased only when broadband-filtered noise was centered on or above the calls' dominant or fundamental frequency. This indicates that ambient noise modifies the associated acoustic parameters of the Lombard-effect, such as spectro-temporal features, independently and could significantly affect the bat's ability to detect and locate targets. Our findings may be of significance in evaluating the impact of environmental noise on echolocation behavior in bats.

Doppler shift compensation (DSC) behaviour in horseshoe bats is a remarkable example of sensorimotor feedback that stabilizes the echo frequency at the bat's optimum hearing range regardless of motion-induced frequency shifts in the echoes. Searching for a related neural interface, the nucleus of the central acoustic tract (NCAT) was investigated in the echolocating horseshoe bat, Rhinolophus rouxi, using various neurophysiological and tracer methods. The NCAT receives bilateral auditory input from the cochlear nuclei and sends projections to regions outside the classical acoustic pathway like the pretectal area or the superior colliculus. The binaural input is excitatory from the contralateral and inhibitory from the ipsilateral ear to 53% of the units, and auditory responses were biased to frontal and contralateral directions. The best frequencies of NCAT neurons match a narrow range above the main frequency component of the bat's species-specific echolocation call (62% of the units), and the neurons exhibit extremely sharp tuning (Q10dB up to 632). DSC is degraded by unilateral electrical or pharmacological microstimulation of the NCAT, and heavily impaired by unilateral lesion of the region. Altogether, the efferents of the NCAT to prevocal areas, the tuning of its neurons to the DSC-relevant echo frequency range, and the possibility to affect DSC by manipulation of the NCAT, support the assumption that the nucleus plays an important role in audio-vocal control in the horseshoe bat.

Echolocation signals of horseshoe bats (Rhinolophidae) consist of a relatively long component of constant frequency (CF) which is preceded by an initial frequency-modulated (iFM) component and followed by a terminal frequency-modulated (tFM) component. To examine the role of these components in echolocation, four bats were trained to fly from a perch to a landing bar. A dual camera system allowed reconstruction of the flight paths in three dimensions. Echolocation signals were recorded, analyzed, and correlated with the flight behavior of the bats. It was confirmed that during flight the bats compensate the Doppler shifts which are produced by their own flight movement. In free flight they emit per wing beat one single signal of long duration, with little variation in the three signal components. In approach flight the bats reduce pulse duration and interval with decreasing target range. The iFM is not varied with respect to target range, suggesting that this component plays little role in the processing of echolocating a target of interest. The bandwidth of the tFM component is increased while its duration is shortened in proportion to decreasing target range, so that the signal-echo overlap of the FM component is avoided down to a target distance of 15 cm. These concurrent changes suggest that the tFM component is used for ranging. During the last 60 cm of the approach the bats compensated for the increase of echo SPL by lowering the emission level of the CF component by 6-9 dB and that of the tFM component by 9-11 dB per halving of range. The specific signal structure of horseshoe bats is discussed as an adaptation for the hunting of fluttering insects in highly cluttered environments.

We previously reported that band-pass filtered noise (BFN, bandwidth 20 kHz) affected the echolocation behavior of horseshoe bats in different ways depending on which frequencies within the bats’ hearing range BFN was centered. We found that call amplitudes only increased when BFN was centered on the dominant frequency of the bats' calls. In contrast, call frequencies were shifted for all BFN stimuli centered on or below the dominant frequency of echolocation calls including when BFN was centered at 20 kHz (BFN20), which is far below the range of the bats’ echolocation call frequencies. In the present study we focused on masking effects of BFN20 and used it as a model stimulus for anthropogenic noise to investigate how this noise affected call parameters of echolocation calls, such as their frequency, amplitude, duration and rate. We show that only call frequencies shifted in response to masking BFN20, whereas all other call parameters tested exhibited no noise-related changes. Our findings provide an empirical basis to quantitatively predict the impact of anthropogenic noise on echolocation behavior in bats. They also emphasize the need for a better understanding of the impact of anthropogenic noise on bioacoustic communication and orientation systems in general.

Prey pursuit by an echolocating bat was studied theoretically and experimentally. First, a mathematical model was proposed to describe the flight dynamics of a bat and a single prey. In this model, the flight angle of the bat was affected by [Formula: see text] angles related to the flight path of the single moving prey, that is, the angle from the bat to the prey and the flight angle of the prey. Numerical simulation showed that the success rate of prey capture was high, when the bat mainly used the angle to the prey to minimize the distance to the prey, and also used the flight angle of the prey to minimize the difference in flight directions of itself and the prey. Second, parameters in the model were estimated according to experimental data obtained from video recordings taken while a Japanese horseshoe bat (Rhinolphus derrumequinum nippon) pursued a moving moth (Goniocraspidum pryeri) in a flight chamber. One of the estimated parameter values, which represents the ratio in the use of the [Formula: see text] angles, was consistent with the optimal value of the numerical simulation. This agreement between the numerical simulation and parameter estimation suggests that a bat chooses an effective flight path for successful prey capture by using the [Formula: see text] angles. Finally, the mathematical model was extended to include a bat and [Formula: see text] prey. Parameter estimation of the extended model based on laboratory experiments revealed the existence of bat's dynamical attention towards [Formula: see text] prey, that is, simultaneous pursuit of [Formula: see text] prey and selective pursuit of respective prey. Thus, our mathematical model contributes not only to quantitative analysis of effective foraging, but also to qualitative evaluation of a bat's dynamical flight strategy during multiple prey pursuit.

Bats of the family Rhinolophidae emit their echolocation calls through their nostrils and feature elaborate noseleaves shaping the directionality of the emissions. The calls of these bats consist of a long constant-frequency component preceded and/or followed by short frequency-modulated sweeps. While Rhinolophidae are known for their physiological specializations for processing the constant frequency part of the calls, previous evidence suggests that the noseleaves of these animals are tuned to the frequencies in the frequency modulated components of the calls. In this paper, we seek further support for this hypothesis by simulating the emission beam pattern of the bat Rhinolophus formosae. Filling the furrows of lancet and removing the basal lappets (i.e., two flaps on the noseleaf) we find that these conspicuous features of the noseleaf focus the emitted energy mostly for frequencies in the frequency-modulated components. Based on the assumption that this component of the call is used by the bats for ranging, we develop a qualitative model to assess the increase in performance due to the furrows and/or the lappets. The model confirms that both structures decrease the ambiguity in selecting relevant targets for ranging. The lappets and the furrows shape the emission beam for different spatial regions and frequency ranges. Therefore, we conclude that the presented evidence is in line with the hypothesis that different parts of the noseleaves of Rhinolophidae are tuned to different frequency ranges with at least some of the most conspicuous ones being tuned to the frequency modulated components of the callsthus yielding strong evidence for the sensory importance of the component.

Echolocating bats can fly without colliding with obstacles by synchronizing acoustic guidance and flight control. To research how bats adapt echolocation as they became familiar with the spaces, we made CF-FM bats (Japanese horseshoe bat) exhibit S-sharp flight repeatedly in a chamber arranged acoustically transparent chains. We found (1) flight path was changed continuously to reduce its curvature as the bats repeated the flights, resulting that the maximum flight speed increased from 2.7 to 3.0 m/s in the 1st trial to 3.2 to 3.8 m/s in the 12th (last) trial for all three bats, (2) pulse emission was reduced by 45% (66/148 pulses) from the first to the last trial, (3) they shifted the pulse direction dynamically relative to their flight direction in the 1st flight whereas the pulse direction was shifted smoothly, the bats emitted more intense pulses toward the intended flight direction as they became familiar with the space. When acrylic boards were arranged, the bats could not pass through the obstacle ...

Echolocation sounds of Rhinolophus ferrumequinum nippon as they approached a fluttering moth (Goniocraspidum pryeri) were investigated using an on-board telemetry microphone (Telemike). In 40% of the successful moth-capture flights, the moth exhibited distinctive evasive flight behavior, but the bat pursued the moth by following its flight path. When the distance to the moth was approximately 3-4 m, the bats increased the duration of the pulses to 65-95 ms, which is 2-3 times longer than those during landing flight (30-40 ms). The mean of 5.8 long pulses were emitted before the final buzz phase of moth capture, without strengthening the sound pressure level. The mean duration of long pulses (79.9 ± 7.9 ms) corresponded to three times the fluttering period of G. pryeri (26.5 × 3 = 79.5 ms). These findings indicate that the bats adjust the pulse duration to increase the number of temporal repetitions of fluttering information rather than to produce more intense sonar sounds to receive fine insect echoes. The bats exhibited Doppler-shift compensation for echoes returning from large static objects ahead, but not for echoes from target moths, even though the bats were focused on capturing the moths. Furthermore, the echoes of the Telemike recordings from target moths showed spectral glints of approximately 1-1.5 kHz caused by the fluttering of the moths but not amplitude glints because of the highly acoustical attenuation of ultrasound in the air, suggesting that spectral information may be more robust than amplitude information in echoes during moth capturing flight.

The greater horseshoe bat, Rhinolophus ferrumequinum, is a model species in echolocation studies, and emits calls containing long constant-frequency (CF) components. The bats have auditory systems tuned sharply to frequencies close to the resting CF (RF) values. Call frequency and neural processing are both flexible within individual bats which use this mode of echolocation. The simple structure of the calls makes them ideal for sonographic analysis. Here, in a large-scale and long-term analysis of changes in the vocalizations of bats we show that: (i) the calls of R. ferrumequinum aged 1-28 years vary seasonally and over a lifetime in a predictable manner; and (ii) an infant's RF is at least partly determined by the RF of its mother. We consider the relative importance of genetic and learning factors upon the correlation between RFS of mothers and offspring.

This study describes variation patterns in the constant frequency of echolocation calls emitted at rest and when not flying (“resting frequency” RF) of the least horseshoe bat, Rhinolophus pusillus, on a broad geographical scale and in response to local climatic variables. Significant differences in RF were observed among populations throughout the species range in Mainland China, and this variation was positively and significantly related to climate conditions, especially environmental humidity, but the variability was only weakly associated with geographical distance. Sex dimorphism in the RF of R. pusillus may imply that female and male might keep their frequencies within a narrow range for sex recognition. Moreover, bats adjusted resting frequency to humidity, which may imply partitioning diet by prey size or the influence of rainfall noise. The results indicate that bats adjust echolocation call frequency to adapt to environmental conditions. Therefore, environmental selection shape the diversity of echolocation call structure of R. pusillus in geographically separated populations, and conservation efforts should focus on changes in local climate and effects of environmental noise.

We investigated intraspecific variation in echolocation calls of the Cape horseshoe bat, Rhinolophus capensis, by comparing echolocation and associated morphological parameters among individuals from three populations of this species. The populations were situated in the center and at the western and eastern limits of the distribution of R. capensis. The latter two populations were situated in ecotones between vegetation biomes. Ecotone populations deviated slightly from the allometric relationship between body size and peak frequency for the genus, and there was no relationship between these variables within R. capensis. Nasal chamber length was the best predictor of peak frequency but not correlated with body size. The evolution of echolocation thus appears to have been uncoupled from body size in R. capensis. Furthermore, females used higher frequencies than males, which imply a potential social role for peak frequency. The differences in peak frequency may have originated from random founder effects and then compounded by genetic drift and/or natural selection. The latter may have acted directly on peak frequency altering skull parameters involved in echolocation independently of body size, resulting in the evolution of local acoustic signatures.

The origin and maintenance of intraspecific variation in vocal signals is important for population divergence and speciation. Where vocalizations are transmitted by vertical cultural inheritance, similarity will reflect co-ancestry, and thus vocal divergence should reflect genetic structure. Horseshoe bats are characterized by echolocation calls dominated by a constant frequency component that is partly determined by maternal imprinting. Although previous studies showed that constant frequency calls are also influenced by some non-genetic factors, it is not known how frequency relates to genetic structure. To test this, we related constant frequency variation to genetic and non-genetic variables in the Formosan lesser horseshoe bat (Rhinolophus monoceros). Recordings of bats from across Taiwan revealed that females called at higher frequencies than males; however, we found no effect of environmental or morphological factors on call frequency. By comparison, variation showed clear population structure, with frequencies lower in the centre and east, and higher in the north and south. Within these regions, frequency divergence was directional and correlated with geographical distance, suggesting that call frequencies are subject to cultural drift. However, microsatellite clustering analysis showed that broad differences in constant frequency among populations corresponded to discontinuities in allele frequencies resulting from vicariant events. Our results provide evidence that the processes shaping genetic subdivision have concomitant consequences for divergence in echolocation call frequency.

The biosonar system of bats utilizes physical baffle shapes around the sites of ultrasound emission for diffraction-based beam forming. Among these shapes, some extreme cases have evolved that include a long noseleaf protrusion (sella) in a species of horseshoe bat. We have evaluated the acoustic cost function associated with sella length with a computational physics approach and found that the extreme length can be predicted accurately from a fiducial point on this function. This suggests that some extreme cases of biological morphology can be explained from their physical function alone. European Union (CILIA project) Natural Science Foundation of China 10774092 Shandong University (Taishan Fund)

Responses of auditory neurons to complex stimuli were recorded in the dorsal belt region of the auditory cortex of two taxonomically unrelated bat species, Rhinolophus rouxi and Pteronotus parnellii parnellii, both showing Doppler shift compensation behaviour. As in P.p.parnellii (Suga et al., J. Neurophysiol., 49, 1573 - 1626, 1983), cortical neurons of R.rouxi show facilitated responses to pairs of pure tones or frequency modulations. Best frequencies for the two components lie near the first and second harmonic of the echolocation call but are in most cases not harmonically related. Neurons facilitated by pairs of pure tones show little dependence on the delay between the stimuli, whereas pairs of frequency modulations evoke best facilitated responses at distinct best delays between 1 and 10 ms. Facilitated neurons are found in distinct portions of the dorsal cortical belt region, with a segregation of facilitated neurons responding to pure tones and to frequency modulations. Non-facilitated neurons are found throughout the field. Neurons are topographically aligned with increasing best delays along a rostrocaudal axis. The best delays between 2 and 4 ms are largely overrepresented numerically, and occupy approximately 56% of the cortical area containing facilitated neurons. A functional interpretation of the large overrepresentation of best delays approximately 3 ms is proposed. Facilitated neurons are located almost entirely within layer V of the dorsal field.

The present study examines duration-sensitive neurons in the inferior colliculus (IC) of the least horseshoe bat, Rhinolophus pusillus, from China. In contrast to other bat species tested for duration selectivity so far, echolocation pulses emitted by horseshoe bats are generally longer and composed of a long constant-frequency (CF) component followed by a short downward frequency-modulated (FM) sweep (CF-FM pulse). We used combined CF-FM pulses to analyze the differential effects that these two pulse components had on the duration tuning in neurons of the horseshoe bat's IC. Consistent with results from other mammals, duration-sensitive neurons found in the least horseshoe bat fall into three main classes: short-pass, band-pass, and long-pass. Using a CF stimulus alone, 54% (51/95) of all IC neurons showed at least one form of duration selectivity at one or more stimulus intensities. In 65 of the 95 IC neurons tested with CF pulses, we were also able to test their duration selectivity for a combined CF-FM pulse, which increased the ratio of duration-sensitive neurons to 66% (43/65). Seven to 15 neurons that failed to show duration tuning for CF bursts became duration sensitive for CF-FM pulses, with most of them exhibiting short-pass (depending on stimulus intensity, between 4 and 8 neurons) or band-pass tuning (1–3 neurons). Increasing stimulus intensities did not affect the duration tuning in 53% (23/43) of duration-sensitive neurons for CF bursts and in about 26% (7/27) for CF-FM stimuli. In the remaining neurons, increasing sound levels generally reduced the ratio of duration-sensitive neurons to 33% for CF and 37% for CF-FM stimulation. In those that remained duration sensitive, louder CF bursts shortened best durations in band-pass neurons and cutoff durations in short- and long-pass neurons, whereas louder CF-FM stimuli reduced the cutoff durations only in short-pass neurons. Bandwidths of band-pass neurons were not significantly affected by any stimulus configuration, with only a slight trend for increasing bandwidths for louder CF bursts (but not CF-FM stimuli). Best durations and cutoff durations reached higher values than those in the other bat species examined so far and roughly match the longer durations of echolocation pulses emitted by horseshoe bats. Therefore presentation of a CF-FM stimulus improved the duration sensitivity in IC neurons by increasing the ratio of duration-tuned neurons and making them less susceptible to changes in signal intensity.

The Okinawan least horseshoe bat, Rhinolophus pumilus, is a cave-dwelling species endemic to the central and southern Ryukyus, Japan. We analyzed variation in the constant frequency (CF) of the echolocation call and in forearm length (FAL) of this species on Okinawa-jima Island on the basis of data for 479 individuals from 11 caves scattered over the island. CF values in samples from six caves, all located in the southwestern half of Okinawa-jima, were significantly higher than those in samples from five caves in the northeastern half of the island. Also, FAL was significantly greater in the latter group than in the former group, although the ranges of variation in this character substantially overlapped between the two groups. These results suggest substantial differentiation between R. pumilus populations on Okinawa-jima. The implications of our findings for the conservation of this endangered bat species are briefly discussed.

Considering neurobiological aspects of visual system in birds and particularly in owls, echolocation in horseshoe bat and well known visual system in primates/humans we present their common features that concerns content scanning capabilities. In that we see various fulfilments of the same evolutionary concept in quite different ecology frames. Proper comparison is given, showing how sophisticated are certain ecologically determined features of particular scanning process, and its relation to the phenomenon of attention, whatever how highly embedded in brain functions. We gave the explanation of neurobiological background of such mechanism.

Whereas echolocation in horseshoe bats is well studied, virtually nothing is known about characteristics and function of their communication calls. Therefore, the communication calls produced by a group of captive adult greater horseshoe bats were recorded during various social interactions in a free-flight facility. Analysis revealed that this species exhibited an amazingly rich repertoire of vocalizations varying in numerous spectro-temporal aspects. Calls were classified into 17 syllable types (ten simple syllables and seven composites). Syllables were combined into six types of simple phrases and four combination phrases. The majority of syllables had durations of more than 100 ms with multiple harmonics and fundamental frequencies usually above 20 kHz, although some of them were also audible to humans. Preliminary behavioral observations indicated that many calls were emitted during direct interaction with and in response to social calls from conspecifics without requiring physical contact. Some echolocation-like vocalizations also appeared to clearly serve a communication role. These results not only shed light upon a so far widely neglected aspect of horseshoe bat vocalizations, but also provide the basis for future studies on the neural control of the production of communicative vocalizations in contrast to the production of echolocation pulse sequences.

The 71 species of horseshoe bat (genus Rhinolophus) use echolocation calls with long constant-frequency (CF) components to detect and localize fluttering insects which they seize in aerial captures or glean from foliage. Here we describe ground-gleaning as an additional prey-capture strategy for horseshoe bats. This study presents the first record and experimental evidence for ground-gleaning in the little-studied Blasius’ horseshoe bat (Rhinolophus blasii). The gleaning bouts in a flight tent included landing, quadrupedal walking and take-off from the ground. The bats emitted echolocation calls continuously during all phases of prey capture. Both spontaneously and in a choice experiment, all six individuals attacked only fluttering insects and never motionless prey. These data suggest that R. blasii performs ground-gleaning largely by relying on the same prey-detection strategy and echolocation behaviour that it and other horseshoe bats use for aerial hawking. We also studied the Mediterranean horseshoe bat (R. euryale) in the flight tent. All four individuals never gleaned prey from the ground, though they appeared to be well able to detect fluttering moths on the ground. It is not known yet whether ground-gleaning plays a role in Mehely’s horseshoe bat (R. mehelyi). In a performance test, we measured the ability of these three European species of “middle-sized” horseshoe bats (R. euryale, R. mehelyi and R. blasii) to take-off from the ground. All were able to take flight even in a confined space; i.e. the willingness to ground-glean in R. blasii is not related to a superior take-off performance. In contrast to ground-gleaning bats of other phylogenetic lineages, R. blasii appears not to be a specialist, but rather shows a remarkable behavioural flexibility in prey-capture strategies and abilities. We suggest that the key innovation of CF echolocation paired with behavioural flexibility in foraging strategies might explain the evolutionary success of Rhinolophus as the second largest genus of bat.

Evolutionary divergence between species is facilitated by ecological shifts, and divergence is particularly rapid when such shifts also promote assortative mating1,2,3. Horseshoe bats are a diverse Old World family (Rhinolophidae) that have undergone a rapid radiation in the past 5 million years4. These insectivorous bats use a predominantly pure-tone echolocation call matched to an auditory fovea (an over-representation of the pure-tone frequency in the cochlea and inferior colliculus5,6) to detect the minute changes in echo amplitude and frequency generated when an insect flutters its wings7. The emitted signal is the accentuated second harmonic of a series in which the fundamental and remaining harmonics are filtered out8. Here we show that three distinct, sympatric size morphs of the large-eared horseshoe bat (Rhinolophus philippinensis) echolocate at different harmonics of the same fundamental frequency. These morphs have undergone recent genetic divergence, and this process has occurred in parallel more than once9. We suggest that switching harmonics creates a discontinuity in the bats' perception of available prey that can initiate disruptive selection1. Moreover, because call frequency in horseshoe bats has a dual function in resource acquisition and communication, ecological selection on frequency might lead to assortative mating and ultimately reproductive isolation and speciation, regardless of external barriers to gene flow1,2,3.