Hearing sensitivity: An underlying mechanism for niche differentiation in gleaning bats.

Tropical ecosystems are known for high species diversity. Adaptations permitting niche differentiation enable species to coexist. Historically, research focused primarily on morphological and behavioral adaptations for foraging, roosting, and other basic ecological factors. Another important factor, however, is differences in sensory capabilities. So far, studies mainly have focused on the output of behavioral strategies of predators and their prey preference. Understanding the coexistence of different foraging strategies, however, requires understanding underlying cognitive and neural mechanisms. In this study, we investigate hearing in bats and how it shapes bat species coexistence. We present the hearing thresholds and echolocation calls of 12 different gleaning bats from the ecologically diverse Phyllostomid family. We measured their auditory brainstem responses to assess their hearing sensitivity. The audiograms of these species had similar overall shapes but differed substantially for frequencies below 9 kHz and in the frequency range of their echolocation calls. Our results suggest that differences among bats in hearing abilities contribute to the diversity in foraging strategies of gleaning bats. We argue that differences in auditory sensitivity could be important mechanisms shaping diversity in sensory niches and coexistence of species.

Hearing sensitivity and amplitude coding in bats are differentially shaped by echolocation calls and social calls.

Differences in auditory perception between species are influenced by phylogenetic origin and the perceptual challenges imposed by the natural environment, such as detecting prey- or predator-generated sounds and communication signals. Bats are well suited for comparative studies on auditory perception since they predominantly rely on echolocation to perceive the world, while their social calls and most environmental sounds have low frequencies. We tested if hearing sensitivity and stimulus level coding in bats differ between high and low-frequency ranges by measuring auditory brainstem responses (ABRs) of 86 bats belonging to 11 species. In most species, auditory sensitivity was equally good at both high- and low-frequency ranges, while amplitude was more finely coded for higher frequency ranges. Additionally, we conducted a phylogenetic comparative analysis by combining our ABR data with published data on 27 species. Species-specific peaks in hearing sensitivity correlated with peak frequencies of echolocation calls and pup isolation calls, suggesting that changes in hearing sensitivity evolved in response to frequency changes of echolocation and social calls. Overall, our study provides the most comprehensive comparative assessment of bat hearing capacities to date and highlights the evolutionary pressures acting on their sensory perception.

Psychophysical evidence for auditory motion parallax.

Distance is important: From an ecological perspective, knowledge about the distance to either prey or predator is vital. However, the distance of an unknown sound source is particularly difficult to assess, especially in anechoic environments. In vision, changes in perspective resulting from observer motion produce a reliable, consistent, and unambiguous impression of depth known as motion parallax. Here we demonstrate with formal psychophysics that humans can exploit auditory motion parallax, i.e., the change in the dynamic binaural cues elicited by self-motion, to assess the relative depths of two sound sources. Our data show that sensitivity to relative depth is best when subjects move actively; performance deteriorates when subjects are moved by a motion platform or when the sound sources themselves move. This is true even though the dynamic binaural cues elicited by these three types of motion are identical. Our data demonstrate a perceptual strategy to segregate intermittent sound sources in depth and highlight the tight interaction between self-motion and binaural processing that allows assessment of the spatial layout of complex acoustic scenes.

Biosonar spatial resolution along the distance axis: revisiting the clutter interference zone.

Many echolocating bats forage close to vegetation - a chaotic arrangement of prey and foliage where multiple targets are positioned behind one another. Bats excel at determining distance: they measure the delay between the outgoing call and the returning echo. In their auditory cortex, delay-sensitive neurons form a topographic map, suggesting that bats can resolve echoes of multiple targets along the distance axis - a skill crucial for the forage-amongst-foliage scenario. We tested this hypothesis combining an auditory virtual reality with formal psychophysics: we simulated a prey item embedded in two foliage elements, one in front of and one behind the prey. The simulated spacing between 'prey' (target) and 'foliage' (maskers) was defined by the inter-masker delay (IMD). We trained Phyllostomus discolor bats to detect the target in the presence of the maskers, systematically varying both loudness and spacing of the maskers. We show that target detection is impaired when maskers are closely spaced (IMD<1 ms), but remarkably improves when the spacing is increased: the release from masking is approximately 5 dB for intermediate IMDs (1-3 ms) and increases to over 15 dB for large IMDs (≥9 ms). These results are comparable to those from earlier work on the clutter interference zone of bats (Simmons et al., 1988). They suggest that prey would enjoy considerable acoustic protection from closely spaced foliage, but also that the range resolution of bats would let them 'peek into gaps'. Our study puts target ranging into a meaningful context and highlights the limitations of computational topographic maps.

Vocal production learning in the pale spear-nosed bat, Phyllostomus discolor.

Vocal production learning (VPL), or the ability to modify vocalizations through the imitation of sounds, is a rare trait in the animal kingdom. While humans are exceptional vocal learners, few other mammalian species share this trait. Owing to their singular ecology and lifestyle, bats are highly specialized for the precise emission and reception of acoustic signals. This specialization makes them ideal candidates for the study of vocal learning, and several bat species have previously shown evidence supportive of vocal learning. Here we use a sophisticated automated set-up and a contingency training paradigm to explore the vocal learning capacity of pale spear-nosed bats. We show that these bats are capable of directional change of the fundamental frequency of their calls according to an auditory target. With this study, we further highlight the importance of bats for the study of vocal learning and provide evidence for the VPL capacity of the pale spear-nosed bat.

Optic and echo-acoustic flow interact in bats.

Echolocating bats are known to fly and forage in complete darkness, using the echoes of their actively emitted calls to navigate and to detect prey. However, under dim light conditions many bats can also rely on vision. Many flying animals have been shown to navigate by optic flow information and, recently, bats were shown to exploit echo-acoustic flow to navigate through dark habitats. Here, we show for the bat Phyllostomus discolor that, in lighted habitats where self-motion-induced optic flow is strong, optic and echo-acoustic flow interact to guide navigation. Echo-acoustic flow showed a surprisingly strong effect compared with optic flow. We thus demonstrate multimodal interaction between two far-ranging spatial senses, vision and echolocation, available in this combination almost exclusively in bats and toothed whales. Our results highlight the importance of merging information from different sensory systems in a sensory-specialist animal to successfully navigate and hunt under difficult conditions.

The percept of reverberation is not affected by visual room impression in virtual environments.

Humans possess mechanisms to suppress distracting early sound reflections, summarized as the precedence effect. Recent work shows that precedence is affected by visual stimulation. This paper investigates possible effects of visual stimulation on the perception of later reflections, i.e., reverberation. In a highly immersive audio-visual virtual reality environment, subjects were asked to quantify reverberation in conditions where simultaneously presented auditory and visual stimuli either match in room identity, sound source azimuth, and sound source distance, or diverge in one of these aspects. While subjects reliably judged reverberation across acoustic environments, the visual room impression did not affect reverberation estimates.

Human Exploration of Enclosed Spaces through Echolocation.

Some blind humans have developed echolocation, as a method of navigation in space. Echolocation is a truly active sense because subjects analyze echoes of dedicated, self-generated sounds to assess space around them. Using a special virtual space technique, we assess how humans perceive enclosed spaces through echolocation, thereby revealing the interplay between sensory and vocal-motor neural activity while humans perform this task. Sighted subjects were trained to detect small changes in virtual-room size analyzing real-time generated echoes of their vocalizations. Individual differences in performance were related to the type and number of vocalizations produced. We then asked subjects to estimate virtual-room size with either active or passive sounds while measuring their brain activity with fMRI. Subjects were better at estimating room size when actively vocalizing. This was reflected in the hemodynamic activity of vocal-motor cortices, even after individual motor and sensory components were removed. Activity in these areas also varied with perceived room size, although the vocal-motor output was unchanged. In addition, thalamic and auditory-midbrain activity was correlated with perceived room size; a likely result of top-down auditory pathways for human echolocation, comparable with those described in echolocating bats. Our data provide evidence that human echolocation is supported by active sensing, both behaviorally and in terms of brain activity. The neural sensory-motor coupling complements the fundamental acoustic motor-sensory coupling via the environment in echolocation.SIGNIFICANCE STATEMENT Passive listening is the predominant method for examining brain activity during echolocation, the auditory analysis of self-generated sounds. We show that sighted humans perform better when they actively vocalize than during passive listening. Correspondingly, vocal motor and cerebellar activity is greater during active echolocation than vocalization alone. Motor and subcortical auditory brain activity covaries with the auditory percept, although motor output is unchanged. Our results reveal behaviorally relevant neural sensory-motor coupling during echolocation.

Flutter sensitivity in FM bats. Part I: delay modulation.

Echolocating bats measure target distance by the time delay between call and echo. Target movement such as the flutter of insect wings induces delay modulations. Perception of delay modulations has been studied extensively in bats, but only concerning how well bats discriminate flutter frequencies, never with regard to flutter magnitude. We used an auditory virtual reality approach to generate changes in echo delay that were independent of call repetition rate, mimicking fluttering insect wings. We show that in the frequency-modulating (FM) bat Phyllostomus discolor, the sensitivity for modulations in echo delay depends on the rate of the modulation, with bats being most sensitive at modulation rates below 20 Hz and above 50 Hz. The very short duration of their calls compels FM bats to evaluate slow modulations (< about 100 Hz) across entire echo sequences. This makes them susceptible to interference between their own call repetition rate and the modulation rate. We propose that this phenomenon constitutes an echo-acoustic wagon-wheel effect. We further demonstrate how at high modulation rates, flutter sensitivity could be rescued by using spectral and temporal cues introduced by Doppler distortions. Thus, Doppler distortions may play a crucial role in flutter sensitivity in the hundreds of FM species worldwide.

Flutter sensitivity in FM bats. Part II: amplitude modulation.

Bats use echolocation to detect targets such as insect prey. The echolocation call of frequency-modulating bats (FM bats) typically sweeps through a broad range of frequencies within a few milliseconds. The large bandwidth grants the bat high spatial acuity in depicting the target. However, the extremely short call duration and the overall low duty cycle of call emission impair the bat's capability to detect e.g. target movement. Nonetheless, FM bats constitute more than 80% of all echolocating species and are able to navigate and forage in an environment full of moving targets. We used an auditory virtual reality approach to generate changes in echo amplitude reflective of fluttering insect wings independently from other confounding parameters. We show that the FM bat Phyllostomus discolor successfully detected these modulations in echo amplitude and that their performance increased with the rate of the modulation, mimicking faster insect wing-beats. The ability of FM bats to detect amplitude modulations of echoes suggests a release from the trade-off between spatial and temporal acuity and highlights the diversity of selective pressures working on the echolocation system of bats.

Volitional control of social vocalisations and vocal usage learning in bats.

Bats are gregarious, highly vocal animals that possess a broad repertoire of social vocalisations. For in-depth studies of their vocal behaviours, including vocal flexibility and vocal learning, it is necessary to gather repeatable evidence from controlled laboratory experiments on isolated individuals. However, such studies are rare for one simple reason: eliciting social calls in isolation and under operant control is challenging and has rarely been achieved. To overcome this limitation, we designed an automated setup that allows conditioning of social vocalisations in a new context and tracks spectro-temporal changes in the recorded calls over time. Using this setup, we were able to reliably evoke social calls from temporarily isolated lesser spear-nosed bats (Phyllostomus discolor). When we adjusted the call criteria that could result in a food reward, bats responded by adjusting temporal and spectral call parameters. This was achieved without the help of an auditory template or social context to direct the bats. Our results demonstrate vocal flexibility and vocal usage learning in bats. Our setup provides a new paradigm that allows the controlled study of the production and learning of social vocalisations in isolated bats, overcoming limitations that have, until now, prevented in-depth studies of these behaviours.

Fast sensory-motor reactions in echolocating bats to sudden changes during the final buzz and prey intercept.

Echolocation is an active sense enabling bats and toothed whales to orient in darkness through echo returns from their ultrasonic signals. Immediately before prey capture, both bats and whales emit a buzz with such high emission rates (≥ 180 Hz) and overall duration so short that its functional significance remains an enigma. To investigate sensory-motor control during the buzz of the insectivorous bat Myotis daubentonii, we removed prey, suspended in air or on water, before expected capture. The bats responded by shortening their echolocation buzz gradually; the earlier prey was removed down to approximately 100 ms (30 cm) before expected capture, after which the full buzz sequence was emitted both in air and over water. Bats trawling over water also performed the full capture behavior, but in-air capture motions were aborted, even at very late prey removals (<20 ms = 6 cm before expected contact). Thus, neither the buzz nor capture movements are stereotypical, but dynamically adapted based on sensory feedback. The results indicate that echolocation is controlled mainly by acoustic feedback, whereas capture movements are adjusted according to both acoustic and somatosensory feedback, suggesting separate (but coordinated) central motor control of the two behaviors based on multimodal input. Bat echolocation, especially the terminal buzz, provides a unique window to extremely fast decision processes in response to sensory feedback and modulation through attention in a naturally behaving animal.

Echo-acoustic scanning with noseleaf and ears in phyllostomid bats.

The mammalian visual system is highly directional and mammals typically employ rapid eye movements to scan their environment. Both sound emission and hearing in echolocating bats are directional but not much is known about how bats use ear movements and possibly movements of the sound-emitting structures to scan space. Here, we investigated in a tightly controlled behavioural experiment how Phyllostomusdiscolor bats employ their echolocation system while being moved through differently structured environments: we monitored and reconstructed both a close-up of the facial structures in 3D, including the motile noseleaf and outer ears, and the sonar-beam of the bat while it was moved along reflectors. Despite the simple linear movement of the bats in the setup, the bats pointed their beam quite variably in azimuth with a standard deviation of about ±20 deg. This variation arises from yaw-type head rotations. Video analyses show that the bat's noseleaf twitches with every echolocation call. Second, we show that the bat's ears are raised to a rather stereotypical head-centred position with every echolocation call. Surprisingly, P. discolor can adjust the timing and the magnitude of these ear movements to the distance of the reflectors with millisecond precision. Our findings reveal echolocation-specific specialisations as well as general principles of scanning and stabilisation of a directional remote sense. The call-correlated movements of the facial structures may lead to a higher directionality of the echolocation system and may enable the bats to adjust their echo-acoustic gaze to dynamic environments.

The Lombard effect emerges early in young bats: implications for the development of audio-vocal integration.

Auditory feedback plays an important role in vocal learning and, more generally, in fine-tuning the acoustic features of communication signals. So far, only a few studies have assessed the developmental onset of auditory feedback. The Lombard effect, a well-studied audio-vocal phenomenon, refers to an increase in vocal loudness of a subject in response to an increase in background noise. Here, we studied the time course of the Lombard effect in developing bats, Phyllostomus discolor We show that infant bats produced louder vocalizations in noise than in silence at an age of only 2 weeks. In contrast, the infant bats' morphology and vocalizations changed gradually until 2 months of age. Furthermore, we found that the Lombard magnitude, i.e. how much the bats increased their vocal loudness in noise relative to silence, correlated positively with the age of the infant bats. We conclude that the Lombard effect features an early developmental origin, indicating a fast maturation of the underlying neural circuits for audio-vocal feedback.

Dependence of auditory spatial updating on vestibular, proprioceptive, and efference copy signals.

Humans localize sounds by comparing inputs across the two ears, resulting in a head-centered representation of sound-source position. When the head moves, information about head movement must be combined with the head-centered estimate to correctly update the world-centered sound-source position. Spatial updating has been extensively studied in the visual system, but less is known about how head movement signals interact with binaural information during auditory spatial updating. In the current experiments, listeners compared the world-centered azimuthal position of two sound sources presented before and after a head rotation that depended on condition. In the active condition, subjects rotated their head by ∼35° to the left or right, following a pretrained trajectory. In the passive condition, subjects were rotated along the same trajectory in a rotating chair. In the cancellation condition, subjects rotated their head as in the active condition, but the chair was counter-rotated on the basis of head-tracking data such that the head effectively remained fixed in space while the body rotated beneath it. Subjects updated most accurately in the passive condition but erred in the active and cancellation conditions. Performance is interpreted as reflecting the accuracy of perceived head rotation across conditions, which is modeled as a linear combination of proprioceptive/efference copy signals and vestibular signals. Resulting weights suggest that auditory updating is dominated by vestibular signals but with significant contributions from proprioception/efference copy. Overall, results shed light on the interplay of sensory and motor signals that determine the accuracy of auditory spatial updating.

Biomechanical control of vocal plasticity in an echolocating bat.

Many animal species adjust the spectral composition of their acoustic signals to variable environments. However, the physiological foundation of such spectral plasticity is often unclear. The source-filter theory of sound production, initially established for human speech, applies to vocalizations in birds and mammals. According to this theory, adjusting the spectral structure of vocalizations could be achieved by modifying either the laryngeal/syringeal source signal or the vocal tract, which filters the source signal. Here, we show that in pale spear-nosed bats, spectral plasticity induced by moderate level background noise is dominated by the vocal tract rather than the laryngeal source signal. Specifically, we found that with increasing background noise levels, bats consistently decreased the spectral centroid of their echolocation calls up to 3.2 kHz, together with other spectral parameters. In contrast, noise-induced changes in fundamental frequency were small (maximally 0.1 kHz) and were inconsistent across individuals. Changes in spectral centroid did not correlate with changes in fundamental frequency, whereas they correlated negatively with changes in call amplitude. Furthermore, while bats consistently increased call amplitude with increasing noise levels (the Lombard effect), increases in call amplitude typically did not lead to increases in fundamental frequency. In summary, our results suggest that at least to a certain degree echolocating bats are capable of adjusting call amplitude, fundamental frequency and spectral parameters independently.

Echo-acoustic flow affects flight in bats.

Flying animals need to react fast to rapid changes in their environment. Visually guided animals use optic flow, generated by their movement through structured environments. Nocturnal bats cannot make use of optic flow, but rely mostly on echolocation. Here, we show that bats exploit echo-acoustic flow to negotiate flight through narrow passages. Specifically, bats' flight between lateral structures is significantly affected by the echo-acoustic salience of those structures, independent of their physical distance. This is true even though echolocation, unlike vision, provides explicit distance cues. Moreover, the bats reduced the echolocation sound levels in stronger flow, probably to compensate for the increased summary target strength of the lateral reflectors. However, bats did not reduce flight velocity under stronger echo-acoustic flow. Our results demonstrate that sensory flow is a ubiquitous principle for flight guidance, independent of the fundamentally different peripheral representation of flow across the senses of vision and echolocation.

Responses of the Human Inner Ear to Low-Frequency Sound.

The perceptual insensitivity to low frequency (LF) sound in humans has led to an underestimation of the physiological impact of LF exposure on the inner ear. It is known, however, that intense, LF sound causes cyclic changes of indicators of inner ear function after LF stimulus offset, for which the term "Bounce" phenomenon has been coined.Here, we show that the mechanical amplification of hair cells (OHCs) is significantly affected after the presentation of LF sound. First, we show the Bounce phenomenon in slow level changes of quadratic, but not cubic, distortion product otoacoustic emissions (DPOAEs). Second, Bouncing in response to LF sound is seen in slow, oscillating frequency and correlated level changes of spontaneous otoacoustic emissions (SOAEs). Surprisingly, LF sound can induce new SOAEs which can persist for tens of seconds. Further, we show that the Bounce persists under free-field conditions, i.e. without an in-ear probe occluding the auditory meatus. Finally, we show that the Bounce is affected by contralateral acoustic stimulation synchronised to the ipsilateral LF sound. These findings clearly demonstrate that the origin of the Bounce lies in the modulation of cochlear amplifier gain. We conclude that activity changes of OHCs are the source of the Bounce, most likely caused by a temporary disturbance of OHC calcium homeostasis. In the light of these findings, the effects of long-duration, anthropogenic LF sound on the human inner ear require further research.

A novel approach identifies the first transcriptome networks in bats: a new genetic model for vocal communication.

BACKGROUND: Bats are able to employ an astonishingly complex vocal repertoire for navigating their environment and conveying social information. A handful of species also show evidence for vocal learning, an extremely rare ability shared only with humans and few other animals. However, despite their potential for the study of vocal communication, bats remain severely understudied at a molecular level. To address this fundamental gap we performed the first transcriptome profiling and genetic interrogation of molecular networks in the brain of a highly vocal bat species, Phyllostomus discolor. RESULTS: Gene network analysis typically needs large sample sizes for correct clustering, this can be prohibitive where samples are limited, such as in this study. To overcome this, we developed a novel bioinformatics methodology for identifying robust co-expression gene networks using few samples (N=6). Using this approach, we identified tissue-specific functional gene networks from the bat PAG, a brain region fundamental for mammalian vocalisation. The most highly connected network identified represented a cluster of genes involved in glutamatergic synaptic transmission. Glutamatergic receptors play a significant role in vocalisation from the PAG, suggesting that this gene network may be mechanistically important for vocal-motor control in mammals. CONCLUSION: We have developed an innovative approach to cluster co-expressing gene networks and show that it is highly effective in detecting robust functional gene networks with limited sample sizes. Moreover, this work represents the first gene network analysis performed in a bat brain and establishes bats as a novel, tractable model system for understanding the genetics of vocal mammalian communication.

Biosonar navigation above water II: exploiting mirror images.

As in vision, acoustic signals can be reflected by a smooth surface creating an acoustic mirror image. Water bodies represent the only naturally occurring horizontal and acoustically smooth surfaces. Echolocating bats flying over smooth water bodies encounter echo-acoustic mirror images of objects above the surface. Here, we combined an electrophysiological approach with a behavioral experimental paradigm to investigate whether bats can exploit echo-acoustic mirror images for navigation and how these mirrorlike echo-acoustic cues are encoded in their auditory cortex. In an obstacle-avoidance task where the obstacles could only be detected via their echo-acoustic mirror images, most bats spontaneously exploited these cues for navigation. Sonar ensonifications along the bats' flight path revealed conspicuous changes of the reflection patterns with slightly increased target strengths at relatively long echo delays corresponding to the longer acoustic paths from the mirrored obstacles. Recordings of cortical spatiotemporal response maps (STRMs) describe the tuning of a unit across the dimensions of elevation and time. The majority of cortical single and multiunits showed a special spatiotemporal pattern of excitatory areas in their STRM indicating a preference for echoes with (relative to the setup dimensions) long delays and, interestingly, from low elevations. This neural preference could effectively encode a reflection pattern as it would be perceived by an echolocating bat detecting an object mirrored from below. The current study provides both behavioral and neurophysiological evidence that echo-acoustic mirror images can be exploited by bats for obstacle avoidance. This capability effectively supports echo-acoustic navigation in highly cluttered natural habitats.