Noseleaf dynamics during pulse emission in horseshoe bats

Horseshoe bats emit their biosonar pulses nasally and diffract the outgoing ultrasonic waves by conspicuous structures that surrounded the nostrils. Here, we report quantitative experimental data on the motion of a prominent component of these structures, the anterior leaf, using synchronized laser Doppler vibrometry and acoustic recordings in the greater horseshoe bat (Rhinolophus ferrumequinum). The vibrometry data has demonstrated non-random motion patterns in the anterior leaf. In these patterns, the outer rim of the walls of the anterior leaf twitches forward and inwards to decrease the aperture of the noseleaf and increase the curvature of its surfaces. Noseleaf displacements were correlated with the emitted ultrasonic pulses. After their onset, the inward displacements increased monotonically towards their maximum value which was always reached within the duration of the biosonar pulse, typically towards its end. In other words, the anterior leaf's surfaces were moving inwards during most of the pulse. Non-random motions were not present in all recorded pulse trains, but could apparently be switched on or off. Such switches happened between sequences of consecutive pulses but were never observed between individual pulses within a sequence. The amplitudes of the emitted biosonar pulse and accompanying noseleaf movement were not correlated in the analyzed data set. The measured velocities of the noseleaf surface were too small to induce Doppler shifts of a magnitude with a likely significance. However, the displacement amplitudes were significant in comparison with the overall size of the anterior leaf and the sound wavelengths. These results indicate the possibility that horseshoe bats use dynamic sensing principles on the emission side of their biosonar system. Given the already available evidence that such mechanisms exist for biosonar reception, it may be hypothesized that time-variant mechanisms play a pervasive role in the biosonar sensing of horseshoe bats.     

Spectral and temporal gating mechanisms enhance the clutter rejection in the echolocating bat,Rhinolophus rouxi

Summary Doppler shift compensation behaviour in horseshoe bats, Rhinolophus rouxi, was used to test the interference of pure tones and narrow band noise with compensation performance. The distortions in Doppler shift compensation to sinusoidally frequency shifted echoes (modulation frequency: 0.1 Hz, maximum frequency shift: 3 kHz) consisted of a reduced compensation amplitude and/or a shift of the emitted frequency to lower frequencies (Fig. 1).Pure tones at frequencies between 200 and 900 Hz above the bat's resting frequency (RF) disturbed the Doppler shift compensation, with a maximum of intererence between 400 and 550 Hz (Fig. 2). Minimum duration of pure tones for interference was 20 ms and durations above 40 ms were most effective (Fig. 3). Interfering pure tones arriving later than about 10 ms after the onset of the echolocation call showed markedly reduced interference (Fig. 4). Doppler shift compensation was affected by pure tones at the optimum interfering frequency with sound pressure levels down to –48 dB rel the intensity level of the emitted call (Figs. 5, 6).Narrow bandwidth noise (bandwidth from ± 100 Hz to ± 800 Hz) disturbed Doppler shift compensation at carrier frequencies between –250 Hz below and 800 Hz above RF with a maximum of interference between 250 and 500 Hz above resting frequency (Fig. 7). The duration and delay of the noise had similar influences on interference with Doppler shift compensation as did pure tones (Figs. 8, 9). Intensity dependence for noise interference was more variable than for pure tones (-32 dB to -45 dB rel emitted sound pressure level, Fig. 10).The temporal and spectral gating in Doppler shift compensation behaviour is discussed as an effective mechanism for clutter rejection by improving the processing of frequency and amplitude transients in the echoes of horseshoe bats.Abbreviations CF constant frequency - FM frequency modulation - RF resting frequency - SPL sound pressure level     

Ontogenesis of the echolocation system in the rufous horseshoe bat,Rhinolophus rouxi (Audition and vocalization in early postnatal development)

1. The development of vocalization and hearing was studied in Sri Lankan horseshoe bats (Rhinolophus rouxi) during the first postnatal month. The young bats were caught in a nursing colony of rhinolophids in which birth took place within a two week period. 2. The new-born bats emitted isolation calls through the mouth. At the beginning these calls consisted of pure tones with frequencies below 10 kHz (Fig. 1). During the first postnatal week the call frequency increased to about 15 kHz, and the fundamental was augmented by two to four harmonics. No evoked potentials to pure tone stimuli could be elicited in the inferior colliculus of this age group, i.e., auditory processing at the midbrain level was not demonstrable. 3. Evoked potentials were first recorded in the second week, broadly tuned to 15-45 kHz, with a maximum sensitivity between 15-25 kHz. In the course of the second week, however, higher frequencies up to 60 kHz became progressively incorporated into the audiogram (Fig. 3). The fundamental frequency of the multiharmonic isolation calls, emitted strictly through the mouth, increased to about 20 kHz. 4. In the bats' third postnatal week an increased hearing sensitivity (auditory filter) emerged, sharply tuned at frequencies between 57 and 60 kHz (Fig. 4e). The same individuals were also the first to emit long constant frequency echolocation calls through the nostrils (Fig. 4c). The energy of the calls was arranged in harmonic frequency bands with the second harmonic exactly tuned to the auditory filter. These young bats continued to emit isolation calls through the mouth, which were, however, not harmonically related to the echolocation calls (Fig. 4b, d). 5. During the fourth week, both the auditory filter and the matched echolocation pulses (the second harmonic) shifted towards higher frequencies (Fig. 5). During the fifth week the fundamental frequency of the calls was progressively attenuated, and both the second harmonic of the pulses and the auditory filter reached the frequency range typical for adult bats of 73-78 kHz (Fig. 6). 6. The development of audition and vocalization is discussed with regard to possible interactions of both subsystems, and their incorporation into the active orientation system of echolocation.     

Harmonic and frequency structure used for echolocation sound pattern recognition and distance information processing in the rufous horseshoe bat

Summary The rufous horseshoe bat, Rhinolophus rouxi, was trained to discriminate differences in target distance. During the discrimination trials, the bats emitted complex FM/CF/FM pulses containing first harmonic and dominant second harmonic components.Loud free running artificial pulses, simulating the CF/FM part of the natural echolocation components, interfered with the ability of the bat to discriminate target distance. Changes in the frequency or frequency pattern of the artificial pulses resulted in systematic changes in the degree of interference. Interference occurred when artificial CF/FM pulses were presented at frequencies near those of the bat's own first or second harmonic components.These findings suggest that Rhinolophus rouxi uses both the first and second harmonic components of its complex multiharmonic echolocation sound for distance discrimination. For interference to occur, the sound pattern of each harmonic component must contain a CF signal followed by an FM sweep beginning near the frequency of the CF.Abbreviations CF constant frequency - FM frequency modulated     

The evolution of echolocation in bats

Recent molecular phylogenies have changed our perspective on the evolution of echolocation in bats. These phylogenies suggest that certain bats with sophisticated echolocation (e.g. horseshoe bats) share a common ancestry with non-echolocating bats (e.g. Old World fruit bats). One interpretation of these trees presumes that laryngeal echolocation (calls produced in the larynx) probably evolved in the ancestor of all extant bats. Echolocation might have subsequently been lost in Old World fruit bats, only to evolve secondarily (by tongue clicking) in this family. Remarkable acoustic features such as Doppler shift compensation, whispering echolocation and nasal emission of sound each show multiple convergent origins in bats. The extensive adaptive radiation in echolocation call design is shaped largely by ecology, showing how perceptual challenges imposed by the environment can often override phylogenetic constraints.     

Prey-Correlated Spectral Changes in Echolocation Sounds of the Indian False Vampire Megaderma lyra

False Vampires ( Megaderma lyra ) are gleaning bats which emit brief (1 ms) and faint echolocation signals consisting of four harmonics of a shallow frequency downward modulated fundamental (27–19 kHz). The complete signal spans a frequency range from 100 to 19 kHz. In sound recordings from three experimental animals we show that Megaderma lyra shifts the dominant frequency in the echolocation signals in relation to the type of prey offered and to flight style. During roaming flights the mean peak frequency was 63.2 ± 9 kHz (third harmonic). In prey catching flights, peak frequencies were shifted into the fourth harmonic. In flights towards a dish of crawling mealworms, mean peak frequency was raised to 91.2 ± 3.3 kHz. When the bats flew towards living mice the dominant frequency was further increased to 99.8 ± 5.2 kHz, and the second and third harmonic were at least 10 dB fainter or no longer recordable. The additional frequency shift when flying towards mice was not only a consequence of the dominance of the fourth harmonic but also of an additional rise of the fundamental harmonic by nearly 2 kHz. These prey-correlated frequency shifts in echolocation calls showed little variation between the three experimental animals and were reproducible over time. They occurred at or even before take-off of the bats. This is the first report of target-correlated transient adaptations in echolocation calls of any bat species.     

Integration time for short broad band clicks in echolocating FM-bats (Eptesicus fuscus)

Vespertilionid FM-bats (four Eptesicus fuscus and one Vespertilio murinus) were trained in an electronic phantom target simulator to detect synthetic echoes consisting of either one or two clicks. The threshold sound pressure for single clicks was around 47 dB peSPL for all five bats corresponding to a threshold energy of -95 dB re 1 Pa² * s. By varying the interclick interval, T, for double clicks it was shown that the threshold intensity was around — 3 dB relative to the threshold for single clicks at T up to 2.4 ms, indicating perfect power summation of both clicks. A threshold shift of -13.5 dB for a 1 ms train of 20 clicks (0.05 ms interclick interval) confirmed that the bats integrated the power of the stimuli. At T longer than around 2.5 ms the threshold for double clicks was the same as for single clicks. Thus, the bats performed like perfect energy detectors with an integration time of approximately 2.4 ms. This integration time is an order of magnitude shorter than that reported for bats listening passively for pure tones. In our setup the bats emitted sonar signals with durations of 2–3 ms. Hence, the results may indicate that while echolocating the bats integration time is adapted to the duration of the sonar emissions.Abbreviations AGC automatic gain control - FM frequency modulated - peSPL peak equivalent sound pressure level - rms root mean square - SD standard deviation - SE standard error of mean - T interclick interval     

Echo SPL influences the ranging performance of the big brown bat,Eptesicus fuscus

Four bats of the species Eptesicus fuscus were trained in a two-alternative forced-choice procedure to discriminate between two phantom targets that differed in range. The rewarded stimulus was located at a distance of 52.7 cm, while the other unrewarded stimulus was further away. Only one target was presented at a time.In the first experiment we measured the range discrimination performance at an echo SPL of –28 dB relative to the bat's sonar transmission. A 75% correct performance level was arbitrarily defined as threshold and was obtained at a delay difference of 80 s, corresponding to a range difference of 13.8 mm.In the second experiment the delay difference was fixed at 150 s and the echo SPL varied between –8 and –48 dB relative to sonar emissions. The performance of the bats depended on the relative echo SPL. At –28 dB the bats showed the best performance. It deteriorated at an increase of the relative echo SPL to –18 dB and –8 dB. The performance also deteriorated when the relative echo SPL was reduced to –38 dB and –48 dB. Only at low relative echo SPLs did the bats partially compensate for the reduction in echo SPL and increased the SPL of their emitted signals by a few dB.Our results support the hypothesis that neurons exhibiting paradoxical latency shift may be involved in encoding target range. This hypothesis predicts a decrease in performance at high echo SPLs as we found it in our experiments. The observed reduction in performance at very low echo SPLs may be due to a decrease in S/N ratio.     

The Auditory Mechanics of the Outer Ear of the Bush Cricket: A Numerical Approach

Bush crickets have tympanal ears located in the forelegs. Their ears are elaborate, as they have outer-, middle-, and inner-ear components. The outer ear comprises an air-filled tube derived from the respiratory trachea, the acoustic trachea (AT), which transfers sound from the mesothoracic acoustic spiracle to the internal side of the ear drums in the legs. A key feature of the AT is its capacity to reduce the velocity of sound propagation and alter the acoustic driving forces of the tympanum (the ear drum), producing differences in sound pressure and time between the left and right sides, therefore aiding the directional hearing of the animal. It has been demonstrated experimentally that the tracheal sound transmission generates a gain of ∼15 dB and a propagation velocity of 255 ms⁻¹, an approximately 25% reduction from free-field propagation. However, the mechanism responsible for this change in sound pressure level and velocity remains elusive. In this study, we investigate the mechanical processes behind the sound pressure gain in the AT by numerically modeling the tracheal acoustic behavior using the finite-element method and real three-dimensional geometries of the tracheae of the bush cricket Copiphora gorgonensis. Taking into account the thermoviscous acoustic-shell interaction on the propagation of sound, we analyze the effects of the horn-shaped domain, material properties of the tracheal wall, and the thermal processes on the change in sound pressure level in the AT. Through the numerical results obtained, it is discerned that the tracheal geometry is the main factor contributing to the observed pressure gain.     

Prestin Shows Divergent Evolution Between Constant Frequency Echolocating Bats

The gene Prestin encodes a motor protein that is thought to confer the high-frequency sensitivity and selectivity that characterizes the mammalian auditory system. Recent research shows that the Prestin gene has undergone a burst of positive selection on the ancestral branch of the Old World horseshoe and leaf-nosed bats (Rhinolophidae and Hipposideridae, respectively), and also on the branch leading to echolocating cetaceans. Moreover, these two groups share a large number of convergent amino acid sequence replacements. Horseshoe and leaf-nosed bats exhibit narrowband echolocation, in which the emitted calls are based on the second harmonic of a predominantly constant frequency (CF) component, the frequency of which is also over-represented in the cochlea. This highly specialized form of echolocation has also evolved independently in the neotropical Parnell's mustached bat (Pteronotus parnellii). To test whether the convergent evolution of CF echolocation between lineages has arisen from common changes in the Prestin gene, we sequenced the Prestin coding region (~2,212?bp, >99% coverage) in P. parnellii and several related species that use broadband echolocation calls. Our reconstructed Prestin gene tree and amino acid tree showed that P. parnellii did not group together with Old World horseshoe and leaf-nosed bats, but rather clustered within its true sister species. Comparisons of sequences confirmed that P. parnellii shared most amino acid changes with its congeners, and we found no evidence of positive selection in the branch leading to the genus of Pteronotus. Our result suggests that the adaptive changes seen in Prestin in horseshoe and leaf-nosed bats are not necessary for CF echolocation in P. parnellii.     

Out of phase: relevance of the medial septum for directional hearing and phonotaxis in the natural habitat of field crickets

A modified tracheal system is the anatomical basis for a pressure difference receiver in field crickets, where sound has access to the inner and outer side of the tympanum of the ear in the forelegs. A thin septum in the midline of a connecting trachea coupling both ears is regarded to be important in producing frequency-dependent interaural intensity differences (IIDs) for sound localization. However, the fundamental role of the septum in directional hearing has recently been challenged by the finding that the localization ability is ensured even with a perforated septum, at least under controlled laboratory conditions. Here, we investigated the influence of the medial septum on phonotaxis of female Gryllus bimaculatus under natural conditions. Surprisingly, even with a perforated septum, females reliably tracked a male calling song in the field. Although reduced by 5.2 dB, IIDs still averaged at 7.9 dB and provided a reliable proximate basis for the observed behavioural performance of operated females in the field. In contrast, in the closely related species Gryllus campestris the same septum perforation caused a dramatic decline in IIDs over all frequencies tested. We discuss this discrepancy with respect to a difference in the phenotype of their tracheal systems.     

Vocal communication in adult greater horseshoe bats, Rhinolophus ferrumequinum

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.     

Adaptive behavior for texture discrimination by the free-flying big brown bat, <Emphasis Type="Italic">Eptesicus fuscus</Emphasis>

This study examined behavioral strategies for texture discrimination by echolocation in free-flying bats. Big brown bats, Eptesicus fuscus, were trained to discriminate a smooth 16 mm diameter object (S+) from a size-matched textured object (S−), both of which were tethered in random locations in a flight room. The bat’s three-dimensional flight path was reconstructed using stereo images from high-speed video recordings, and the bat’s sonar vocalizations were recorded for each trial and analyzed off-line. A microphone array permitted reconstruction of the sonar beam pattern, allowing us to study the bat’s directional gaze and inspection of the objects. Bats learned the discrimination, but performance varied with S−. In acoustic studies of the objects, the S+ and S− stimuli were ensonified with frequency-modulated sonar pulses. Mean intensity differences between S+ and S− were within 4 dB. Performance data, combined with analyses of echo recordings, suggest that the big brown bat listens to changes in sound spectra from echo to echo to discriminate between objects. Bats adapted their sonar calls as they inspected the stimuli, and their sonar behavior resembled that of animals foraging for insects. Analysis of sonar beam-directing behavior in certain trials clearly showed that the bat sequentially inspected S+ and S−.     

Spatial unmasking in the echolocating Big Brown Bat, <Emphasis Type="Italic">Eptesicus fuscus</Emphasis>

Masking affects the ability of echolocating bats to detect a target in the presence of clutter targets. It can be reduced by spatially separating the targets. Spatial unmasking was measured in a two-alternative-forced-choice detection experiment with four Big Brown Bats detecting a wire at 1 m distance. Depth dependent spatial unmasking was investigated by the bats detecting a wire with a diameter of 1.2 mm in front of a masker with a threshold distance of 11 cm behind the wire. For angle dependent spatial unmasking the masker was turned laterally, starting from its threshold position at 11 cm. With increasing masker angles the bats could detect thinner wires with diameters decreasing from 1.2 mm (target strength −36.8 dB) at 0° to 0.2 mm (target strength −63.0 dB) at 22°. Without masker, the bats detected wire diameters of 0.16 mm (target strength −66.2 dB), reached with masker positions beyond 23° (complete masking release). Analysis of the sonar signals indicated strategies in the echolocation behavior. The bats enhanced the second harmonics of their signals. This may improve the spatial separation between wire and masker due to frequency-dependent directionality increase of sound emission and echo reception.     

Localization of Mycoplasma pulmonis in mice.

Localization of Mycoplasma pulmonis was examined in mice infected by direct contact with previously infected mice. After contact with infected animals, the organisms were shown to become detectable first in the nasal and oral cavities and trachea on the next day, and then they were recovered from the middle ear and brain after 3 and 4 days, respectively. After 7 days of contact, isolation rates retained to be 100% in the nasal, oral and tracheal cavities, while 95% in the middle ear and brain, 25% in the lung and 20% in the vagina and uterus. The number of colonies was the most numerous from the nasal, uterine and vaginal cavities, followed by the trachea, middle ear, oral cavity, brain and lung in this order, except for a few mice having pneumonic lesions giving a large number of the organisms. The isolation rates with these organs were not changed even after 6 weeks of contact and organisms were never detected from the liver, spleen, kidney and heart. Mice from a naturally infected breeding colony showed similar finding to those sacrificed after 6 weeks of experimental contact.     

The tettigoniid (Orthoptera : Tettigoniidae) ear: Multiple functions and structural diversity

The ears of bushcrickets (Orthoptera : Tettigoniidae) consist of a pair of membranes situated beneath the knee of the foretibia. These membranes, or tympana, are backed by a tracheal system dedicated to sound reception. In most species, a trachea opens through a hole, the auditory spiracle, in the pleuron of the prothorax adjacent to the ventilatory spiracle. Experimental evidence clearly shows that in those species possessing such an opening, sound entering through this route is amplified by the shape of the trachea, or at least is modified through resonances created by its tracheal cavities or tubes. In some species the external surface of the tympanic membrane is surrounded by cuticular folds, which may be formed into small resonating cavities or sound guiding pinnae. In other species the tympana are naked. Sound interacts with the external surface of the tympana, either through these cuticular cavities or directly at the surface of the membrane. As with auditory tracheae, the shape of these cavities may modify sound through resonances. This review examines the role of both external and internal sound-guides in sound reception. It discusses how different species may utilize different sound receiving systems and suggests how, in some cases, both external and internal routes may respond to different frequency bands. The maximum sensitivity of the ear invariably coincides with the male's call carrier frequency. Two Australian species of tettigoniid where the ear is clearly not tuned to the male's call are described. The biology of one undescribed species belonging to the subfamily, Zaprochilinae, where not only is the ear untuned, but there is marked sexual dimorphism in ear structure, is elaborated on. The review concludes with an examination of the routes selection may have taken to arrive at the extreme morphologies present in the Tettigoniidae.     

The role of echolocation in the hunting of terrestrial prey – new evidence for an underestimated strategy in the gleaning bat, Megaderma lyra

The observation that gleaning bats detect prey by its noises, together with difficulties in recording their faint sonar calls, have led some authors to conclude that gleaning bats may not use echolocation in certain hunting situations. In particular, it is conjectured that echolocation plays no role in the classification and tracking of prey. In the present study, we show that the gleaning bat, Megaderma lyra, is able to find silent and motionless prey on the ground. The significance of sonar for catching a variety of terrestrial prey is established in a standardized situation. Sonar calls were found to be emitted during all stages, i.e. approach, hovering above the prey, and return to the roost, of every hunting flight. The harmonic pattern of the calls differed significantly between these stages, calls with three or more prominent components prevailing during hovering. Bats identified prey and rejected dummies while hovering above them. During this stage, increased call rates and reduced call durations were found. Echolocation activity during, and the duration of, the hovering phase depended on prey type, in particular on prey movement. The prey-dependent shifts in sonar activity, the broadband call structure with an emphasis on higher harmonics, and a systematic shift of the calls' peak frequencies during hovering, are discussed as adaptations to identifying prey by sonar.     

Fine control of call frequency by horseshoe bats

The auditory system of horseshoe bats is narrowly tuned to the sound of their own echoes. During flight these bats continuously adjust the frequency of their echolocation calls to compensate for Doppler-effects in the returning echo. Horseshoe bats can accurately compensate for changes in echo frequency up to 5 kHz, but they do so through a sequence of small, temporally-independent, step changes in call frequency. The relationship between an echo's frequency and its subsequent impact on the frequency of the very next call is fundamental to how Doppler-shift compensation behavior works. We analyzed how horseshoe bats control call frequency by measuring the changes occurring between many successive pairs of calls during Doppler-shift compensation and relating the magnitude of these changes to the frequency of each intervening echo. The results indicate that Doppler-shift compensation is mediated by a pair of (echo)frequency-specific sigmoidal functions characterized by a threshold, a slope, and an upper limit to the maximum change in frequency that may occur between successive calls. The exact values of these parameters necessarily reflect properties of the underlying neural circuitry of Doppler-shift compensation and the motor control of vocalization, and provide insight into how neural feedback can accommodate the need for speed without sacrificing stability.     

Lancet Dynamics in Greater Horseshoe Bats,Rhinolophus ferrumequinum

Echolocating greater horseshoe bats (Rhinolophus ferrumequinum) emit their biosonar pulses nasally, through nostrils surrounded by fleshy appendages (‘noseleaves’) that diffract the outgoing ultrasonic waves. Movements of one noseleaf part, the lancet, were measured in live bats using two synchronized high speed video cameras with 3D stereo reconstruction, and synchronized with pulse emissions recorded by an ultrasonic microphone. During individual broadcasts, the lancet briefly flicks forward (flexion) and is then restored to its original position. This forward motion lasts tens of milliseconds and increases the curvature of the affected noseleaf surfaces. Approximately 90% of the maximum displacements occurred within the duration of individual pulses, with 70% occurring towards the end. Similar lancet motions were not observed between individual pulses in a sequence of broadcasts. Velocities of the lancet motion were too small to induce Doppler shifts of a biologically-meaningful magnitude, but the maximum displacements were significant in comparison with the overall size of the lancet and the ultrasonic wavelengths. Three finite element models were made from micro-CT scans of the noseleaf post mortem to investigate the acoustic effects of lancet displacement. The broadcast beam shapes were found to be altered substantially by the observed small lancet movements. These findings demonstrate that—in addition to the previously described motions of the anterior leaf and the pinna—horseshoe bat biosonar has a third degree of freedom for fast changes that can happen on the time scale of the emitted pulses or the returning echoes and could provide a dynamic mechanism for the encoding of sensory information.