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Ultrastructure of the horseshoe bat's organ of Corti. I. Scanning electron microscopy.

The organ of Corti of the echolocating horseshoe bat (Rhinolophus rouxi) was investigated with scanning electron microscopy in order to provide a comparison with non-echolocating mammals. Throughout the cochlea of horseshoe bats, each outer hair cell (OHC) possesses three rows of stereocilia and there are no morphological distinctions among the different rows of OHCs. However, there are morphological differences between different regions along the cochlea. In the lower and upper basal turn, the receptor surfaces of OHCs are characterized by extremely wide W-shaped stereocilia bundles and wingshaped cuticular plates. The cuticular plates of OHCs of the middle and outermost rows are arranged parallel to each other. Stereocilia length is only 0.8 microns and there is an exaggerated angle of inclination of the shortest row of stereocilia towards the next taller one. Stereocilia arrangements in the apex of the horseshoe bat's cochlea closely resembles those observed in the midbasal region of the rat cochlea. Inner hair cells (IHC) in the lower basal turn appear specialized. They possess only two rows of stereocilia and only 7-8 stereocilia per row. Their cuticular plates are small and oval and widely separated from one another in the longitudinal direction. IHCs at all other locations possess three and up to four rows of stereocilia and 17-20 stereocilia per row. Their cuticular plates are elongated and closely spaced. The transition from specialized to typical mammalian morphology occurs abruptly (over a distance of about 100-150 microns) at the border between the lower and the upper basal turn. This transition is not accompanied by a change in OHC morphology. In the subsurface of the tectorial membrane, throughout the cochlea, there are distinct imprints of the tallest row of stereocilia of all three rows of OHCs and of the IHCs. Data are discussed in relation to specialized aspects of the cochlear frequency map in horseshoe bats and as possible micromechanical adaptations to ultra-high frequency hearing.

Animals↗

Embryonic staging system for the short-tailed fruit bat, Carollia perspicillata, a model organism for the mammalian order Chiroptera, based upon timed pregnancies in captive-bred animals.

There are approximately 4,800 extant species of mammals that exhibit tremendous morphological, physiological, and developmental diversity. Yet embryonic development has been studied in only a few mammalian species. Among mammals, bats are second only to rodents with regard to species number and habitat range and are the most abundant mammals in undisturbed tropical regions. Bat development, though, remains relatively unstudied. Here, we describe and illustrate a staging series of embryonic development for the short-tailed fruit bat, Carollia perspicillata, based on embryos collected at timed intervals after captive matings. As Carollia can be readily maintained and propagated in captivity and is extremely abundant in the wild, it offers an attractive choice as a chiropteran model organism. This staging system provides a framework for studying Carollia embryogenesis and should prove useful as a guide for embryological studies of other bat species and for comparisons with other orders of mammals.

Animals↗

Mercury vapour lamps interfere with the bat defence of tympanate moths (Operophtera spp.; Geometridae)

Bats often forage near streetlamps, where they catch moths in particular. At least two hypotheses may explain the apparent increase in the availability of moths to bats feeding around streetlamps: (1) the moths become concentrated near the light and therefore more profitable to exploit; and (2) the light interferes with the moths' evasive flight behaviour. We tested the second of these hypotheses by exposing flying male winter moths, Operophtera spp., to bursts of ultrasound (26 kHz, 110 dB sound pressure level) from an electronic source. The light from a 125 W mercury vapour lamp had a quantitative effect on the moths' evasive flight response at close range (within ca 4 m), inhibiting it totally in nearly half (43%, N=125) of the cases. By contrast, moths flying in the surrounding woodland and without interference from the lamp always responded to the sound. Streetlamps of the mercury vapour type (white lamps) thus interfere with the defensive behaviour of moths and presumably increase their vulnerability to echolocating bats. This may have implications for the conservation of both moths and bats. Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗

Immunocytochemical identification of endocrine cells in the pancreas of the fruit bat, Rousettus aegyptiacus.

The fruit bat, Rousettus aegyptiacus, is able to absorb large amounts of glucose in very short periods of time. This ability is partly reflected by the structure of the gastrointestinal tract and pancreas. The aim of this study was to confirm preliminary histochemical studies of the bat pancreas and to identify and quantitate endocrine cells by immunocytochemical techniques in order to understand the ability of the bat to absorb these large amounts of glucose. Endocrine cells were distributed in islets throughout the gland and also occurred as discrete cells in the exocrine ducts. Three-dimensional reconstruction and quantitation showed that the endocrine component of the pancreas occupied 9.1% of the total volume. This is far more than that reported in any other species. Four endocrine cell types were demonstrated. Insulin (beta) cells (51.4%) were located throughout the islet and extended between the glucagon (alpha) cells (30.6%). Somatotostatin (delta) cells (8.8%) and pancreatic polypeptide (PP) cells (17.1%) were irregularly scattered throughout the islets. While the percentage of alpha, beta, and delta cells was similar to that in other species, the percentage of PP cells was higher. The high percentage of endocrine tissue found in the pancreas of the fruit bat may reflect metabolic adaptations involved in the absorption of the high carbohydrate diet of this animal.

Animals↗

A mixed ectoparasite--microparasite model for bat-transmitted rabies.

This paper considers the transmission of rabies to domestic livestock by vampire bats. Vampire bats act as ectoparasites on cattle both by ingesting a small amount of blood every night and by prolonging bleeding by the action of anticoagulant substances in their saliva. In addition to this parasitic action bats may also transmit rabies, inflicting important losses on affected herds by the inevitable mortality due to the infection. We modeled this complex interaction and we also demonstrate that bat control measures are more effective in reducing rabies prevalence and mortality by rabies than cattle vaccination.

Animals↗

Genetic characterization and geographic distribution of rabies virus isolates in Brazil: identification of two reservoirs, dogs and vampire bats.

We analyzed 50 rabies virus samples isolated in Brazil from 12 dogs, 11 cats, 5 vampire bats, 15 cattle, 2 horses, 1 pig, 1 sheep, and 3 humans to investigate the molecular epidemiology of rabies viruses. We sequenced 203 nucleotides on the nucleoprotein gene by direct sequencing of the PCR-amplified products. All the isolates belonged to the genotype 1 and homology of the 203 nucleotides was at least 83.7% among isolates. The main reservoirs were estimated based on the homology of nucleotide sequences. Brazilian rabies virus isolates were clustered into two reservoir groups: dogs and vampire bats. All the dog-related rabies virus isolates showed nucleotide homology greater than 99.0%. Vampire bat-related rabies virus isolates showed nucleotide homology greater than 96.6% and could be further divided into subgroups corresponding to areas where viruses were isolated. These data suggest that circulating rabies variants belong to at least two different genotype clusters in Brazil and that these two clusters are maintained independently among vampire bats and dogs.

Amino Acid Sequence↗

Emerging encephalitogenic viruses: lyssaviruses and henipaviruses transmitted by frugivorous bats.

Three newly recognized encephalitogenic zoonotic viruses spread from fruit bats of the genus Pteropus (order Chiroptera, suborder Megachiroptera) have been recognised over the past decade. These are: Hendra virus, formerly named equine morbillivirus, which was responsible for an outbreak of disease in horses and humans in Brisbane, Australia, in 1994; Australian bat lyssavirus, the cause of a severe acute encephalitis, in 1996; and Nipah virus, the cause of a major outbreak of encephalitis and pulmonary disease in domestic pigs and people in peninsula Malaysia in 1999. Hendra and Nipah viruses have been shown to be the first two members of a new genus, Henipavirus, in the family Paramyxoviridae, subfamily Paramyxovirinae, whereas Australian bat lyssavirus is closely related antigenically to classical rabies virus in the genus Lyssavirus, family Rhabdoviridae, although it can be distinguished on genetic grounds. Hendra and Nipah viruses have neurological and pneumonic tropisms. The first humans and equids with Hendra virus infections died from acute respiratory disease, whereas the second human patient died from an encephalitis. With Nipah virus, the predominant clinical syndrome in humans was encephalitic rather than respiratory, whereas in pigs, the infection was characterised by acute fever with respiratory involvement with or without neurological signs. Two human infections with Australian bat lyssavirus have been reported, the clinical signs of which were consistent with classical rabies infection and included a diffuse, non-suppurative encephalitis. Many important questions remain to be answered regarding modes of transmission, pathogenesis, and geographic range of these viruses.

Animals↗

Ontogenesis of auditory fovea representation in the inferior colliculus of the Sri Lankan rufous horseshoe bat, Rhinolophus rouxi.

This report describes the ontogenesis of tonotopy in the inferior colliculus (IC) of the rufous horseshoe bat (Rhinolophus rouxi). Horseshoe bats are deaf at birth, but consistent tonotopy with a low-to-high frequency gradient from dorsolateral to ventromedial develops from the 2nd up to the 5th week. The representation of the auditory fovea is established in ventro-medio-caudal parts of the IC during the 3rd postnatal week (Fig. 3). Then, a narrow frequency band 5 kHz in width, comprising 16% of the bat's auditory range, captures 50-60 vol% of the IC (Fig. 3c). However, foveal tuning is 10-12 kHz (1/3 octave) lower than in adults; foveal tuning in females (65-68 kHz) is 2-3 kHz higher than in males (62-65 Khz). Thereafter, foveal tuning increases by 1-1.5 kHz per day up to the 5th postnatal week, when the adult hearing range is established (Figs. 4, 5). The increase of sensitivity and of tuning sharpness of single units also follows a low-to-high frequency gradient (Fig. 6). Throughout this development the foveal tuning matches the second harmonic of the echolocation pulses vocalised by these young bats. The results confirm the hypothesis of developmental shifts in the frequency-place code for the foveal high frequency representation in the IC.

Acoustic Stimulation↗

Auditory properties of the superior colliculus in the horseshoe bat, Rhinolophus rouxi.

Auditory response properties were studied in the superior colliculus (SC) of the echolocating horseshoe bat Rhinolophus rouxi, a long CF-FM bat, by the use of stationary, dichotic stimuli. The most striking finding in the horseshoe bat was an enormous overrepresentation of neurons with best frequencies in the range of the constant frequency component of the species specific echolocation call (72% of the auditory neurons). These neurons had response thresholds as low as 0 dB SPL and were narrowly tuned with Q10dB--values up to 400, just as in the nuclei of the primary auditory pathway in this species. This overrepresentation may suggest the importance of the superior colliculus in the context of echolocation behavior. While noise stimuli were not particularly effective, other auditory response properties were similar to those described in other mammals. 65% of the SC neurons in the horseshoe bat responded only to monaural stimulation of one ear, primarily the contralateral one. 32% of the neurons received monaural input from both ears. The proportion of neurons responsive to ipsilateral stimulation (41%) was rather high. Mean response latency was 8.9 ms for contralateral stimulation. A tonotopic organization is lacking, but high-frequency neurons are less frequent in rostral SC.

Acoustic Stimulation↗

Labile cochlear tuning in the mustached bat. II. Concomitant shifts in neural tuning.

Acoustic stimuli near 60 kHz elicit pronounced resonance in the cochlea of the mustached bat (Pteronotus parnellii parnellii). The cochlear resonance frequency (CRF) is near the second harmonic, constant frequency (CF2) component of the bat's biosonar signals. Within narrow bands where CF2 and third harmonic (CF3) echoes are maintained, the cochlea has sharp tuning characteristics that are conserved throughout the central auditory system. The purpose of this study was to examine the effects of temperature-related shifts in the CRF on the tuning properties of neurons in the cochlear nucleus and inferior colliculus. Eighty-two single and multi-unit recordings were characterized in 6 awake bats with chronically implanted cochlear microphonic electrodes. As the CRF changed with body temperature, the tuning curves of neurons sharply tuned to frequencies near the CF2 and CF3 shifted with the CRF in every case, yielding a change in the unit's best frequency. The results show that cochlear tuning is labile in the mustached bat, and that this lability produces tonotopic shifts in the frequency response of central auditory neurons. Furthermore, results provide evidence of shifts in the frequency-to-place code within the sharply tuned CF2 and CF3 regions of the cochlea. In conjunction with the finding that biosonar emission frequency and the CRF shift concomitantly with temperature and flight, it is concluded that the adjustment of biosonar signals accommodates the shifts in cochlear and neural tuning that occur with active echolocation.

Acoustic Stimulation↗

Some comments on the proposed perception of phase and nanosecond time disparities by echolocating bats.

In a series of recent reports, Simmons and his colleagues propose that bats are able to accurately encode the spectral, temporal and phase information of their emitted calls and echoes. The information so encoded is then extracted by the networks of the auditory system with specialized processing. They propose that bats use this information to determine the distance to their target by crosscorrelating the entire structure of the emitted call with the structure of the echo. The idea is that slight deviations in the correlation function can be detected by the bat and the degree of mismatch provides an accurate measure of temporal disparity and hence range. The data in the reports purport to show that bats perceive the phase of ultrasonic signals and that they can resolve temporal disparities of about 10 ns, and thus can distinguish range differences as small as 2 microns. The hypothesis also attempts to explain how a variety of acoustic cues are processed and represented in the auditory system and how they are combined to form a unitary percept of space and fine structure. The theory incorporates some time honored processes of extracting information, such as crosscorrelations. The implications of the hypothesis, however, go far beyond a theory of neural processing and representation of information by ensembles of cells. The hypothesis requires some remarkable abilities, such as the phase coding of ultrasonic signals and a temporal acuity on the order of 10 ns. These features have never been seen in any neurophysiological study of any animal nor has its existence been implied in behavioral studies of other animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Evidence for perception of fine echo delay and phase by the FM bat, Eptesicus fuscus.

The big brown bat, Eptesicus fuscus, can perceive small changes in the delay of FM sonar echoes and shifts in echo phase, which interact with delay. Using spectral cues caused by interference, Eptesicus also can perceive the individual delays of two overlapping FM echoes at small delay separations. These results have been criticized as due to spectral artifacts caused by overlap between stimulus echoes and extraneous sounds (Pollak 1993). However, no amplitude or spectral variations larger than 0.05 dB accompany delay or phase changes produced by the electronic apparatus. No reverberation falls in the narrow span of delays required to produce the bat's performance curve from echo interference cues. Consistent differences in the durations of sonar sounds for 6 bats that perform the same in the experiments demonstrate that overlap between stimulus echoes and extraneous echoes is not necessary, and changes in the amount of echo overlap have no effect on performance. Noise-induced random variations in echo spectra outweigh putative spectral artifacts, and deliberately-introduced spectral "artifacts" do not improve performance overall but instead yield new time-frequency images. Amplitude-latency trading of perceived delay, proposed as a demonstration that the latency of neural discharges encodes delay (Pollak et al. 1977), confirms that the bat's fine delay and phase perception depends on a temporal neural code. The perceived delays depend on stimulus delays, not the delays of extraneous sounds. The rejected criticisms are based on physiological results with random-phase FM stimuli which are irrelevant to neural coding of fine echo delay and phase.

Animals↗

Discrimination of two-wavefront echoes by the big brown bat, Eptesicus fuscus: behavioral experiments and receiver simulations.

1. Echolocating bats (Eptesicus fuscus) were trained to discriminate between simulated targets consisting of one or two echo-wavefronts with internal time delays of up to 100 microseconds. Spectral and temporal properties and total signal energy of the targets were evaluated and predictions for performances of bats derived from receiver models were compared with measured performances. 2. Eptesicus fuscus was able to discriminate a one-wavefront target from two-wavefront targets with distinct internal time delays (12 microseconds, 32-40 microseconds and 52-100 microseconds). Performance was not affected by changes in total signal energy. Bats also successfully discriminated between two-wavefront targets with different internal time delays. 3. Performance predicted from differences in total energy between targets did not match the measured performance, indicating that bats did not rely on total echo energy. This finding is also supported by the behavioral data. Performance predicted from spectral and temporal receiver models both matched the measured performance and, therefore, neither one of these models can be favored over the other. 4. The behavioral data suggest that Eptesicus fuscus did not transform echo information into estimates of target range separation and, therefore, did not perceive the two wavefronts of each simulated two-wavefront echo as two separate targets.

Acoustic Stimulation↗

The acoustic role of tracheal chambers and nasal cavities in the production of sonar pulses by the horseshoe bat, Rhinolophus hildebrandti.

The acoustic role of the enlarged, bony, nasal cavities and rigid tracheal chambers in the horseshoe bat, Rhinolophus hildebrandti (Fig. 2) was investigated by determining the effect of their selective filling on the nasally emitted sonar pulse and on the sound traveling backwards down the trachea. Normal sonar signals of this bat contain a long constant frequency component with most energy in the second harmonic at about 48 kHz. The fundamental is typically suppressed 20 to 30 dB below the level of the second harmonic (Fig. 1). None of the experimental manipulations described affected the frequency of the sonar signal fundamental. Filling the dorsal and both lateral tracheal chambers had little effect on the emitted vocalization, but caused the level of the fundamental component in the trachea to increase 15 to 19 dB in most bats (Table 2). When only the dorsal chamber or only the two lateral chambers were filled, the effect was less striking and more variable (Tables 3 and 4), suggesting that the tracheal fundamental is normally suppressed by acoustic interaction between these three cavities. Filling the enlarged dorsal nasal cavities had no effect on the tracheal sound. The effect of this treatment on the nasally emitted sonar pulse was inconsistent. Sometimes the fundamental increased 10 to 12 dB, other times the intensity of all harmonics decreased; in still other cases the second, third or fourth harmonic increased, but the fundamental remained unchanged (Tables 5, 6, and 7). When bats were forced to vocalize through the mouth, by sealing the nostrils, there was a prominent increase in the level of the emitted fundamental (10 to 21 dB) and in the fourth harmonic (6 to 17 dB). In one instance there was also a significant increase in the level of the third harmonic (Tables 8 and 9). The supraglottal tract thus filters the fundamental from the nasally emitted sonar signal, although the role of the inflated nasal cavities in this process is unclear. We conclude that a high glottal impedance acoustically isolates the subglottal from the supraglottal vocal tract. The tracheal chambers do not affect the emitted sonar signal, but may attenuate the fundamental in the trachea and prevent it from being reflected from the lungs back towards the cochlea. It may be important to prevent the reflected fundamental from stimulating the cochlea, via tissue conduction, along multiple indirect pathways which would temporally smear cochlear stimulation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Topographic representation of vocal frequency demonstrated by microstimulation of anterior cingulate cortex in the echolocating bat, Pteronotus parnelli parnelli.

1. A midline region of brain dorsal and anterior to the corpus callosum, presumably anterior cingulate cortex, has been explored for its role in the production of vocalization in the mustached bat, Pteronotus p. parnelli. 2. Vocalizations elicited by microstimulation were virtually indistinguishable from natural biosonar sounds. The spectral content, relative intensity of harmonic components, and durations of emitted pulses are comparable to spontaneous emissions. 3. The frequencies of elicited vocalizations were within the range typically used by the mustached bat during Doppler-shift compensation. The frequency of the second-harmonic constant-frequency component (CF2) covered the range from 57-62 kHz, but was most commonly emitted at frequencies of 59-61 kHz. 4. An increase in the frequency of vocalizations over a number of consecutive pulses towards a steady-state plateau is evident in both spontaneous vocalizations and emissions elicited by microstimulation just above threshold. Increasing the stimulus intensity caused the frequency of emissions to approach the steady state more rapidly. 5. The anterior cingulate cortex appears to be organized topographically for increasing frequency of elicited biosonar sounds along a rostrocaudal axis. The area from which biosonar emissions were elicited was overrepresented for a 2 kHz band of frequencies just below the bats' CF2 resting frequency. Audible vocalizations with a complex spectrum resembling social cries can also be elicited by microstimulation, but only in an area that is adjacent and posterior to the biosonar region. 6. Some examples of both elicited and spontaneous vocalizations contained a relative intensity pattern of the harmonic components which deviated from the typical pattern. This suggests that mustached bats are capable of actively altering the spectrum of their pulses to subserve different tasks in echolocation.

Animals↗

[Comparative electron microscope studies on the kinetoplast morphology of bat-trypanosomes and Trypanosoma cruzi (author's transl)].

Comparative studies on the ultrastructural morphology of kinetoplasts of trypanosomes belonging to the subgenus Schizotrypanum have been made. Three isolates of Trypanosoma vespertilionis and two strains of Trypanosoma dionisii derived from European bats were tested. Comparison was made also with two isolates from Brasilian bats characterized as T. cruzi and two strains which were derived from two patients suffering from Chagas' disease. In the epimastigote culture form of T. cruzi a typical configuration of kDNA becomes obvious, appearing as a central band at the beginning of cell division. It was investigated whether this morphological character can be used in differentiating between bat-trypanosomes and T. cruzi and on the species characterization of bat-trypanosomes. The central band of kDNA could be demonstrated in all cases of the trypanosomes examined. The special configuration of kDNA makes it possible to distinguish exactly between trypanosomes of the subgenus Schizotrypanum and other trypanosomes. Nevertheless this pecularity alone is not sufficient for characterizing species of that subgenus.

Animals↗

A natural focus of Histoplasma capsulatum var. duboisii is a bat cave.

The natural reservoir of Histoplasma capsulatum var. duboisii, the etiological agent of histoplasmosis duboisii (African histoplasmosis) is not yet known. We report the isolation of H. capsulatum var. duboisii from soil admixed with bat guano and from the intestinal contents of a bat in a sandstone cave in a rural area, Ogbunike in Anambra State of Nigeria. Eight of 45 samples of soil admixed with bat guano yielded H. capsulatum var. duboisii. Of the 35 bats belonging to the species Nycteris hispida and Tadirida pumila examined, only one (N. hispida) yielded this fungus from its intestinal contents. Identification of the isolates as Histoplasma was confirmed by exoantigen tests and by mating with tester strains of H. capsulatum. In vitro conversion to large yeast from suggestive of H. capsulatum var. duboisii was obtained on brain heart infusion agar supplemented with sheep blood and glutamine or cysteine. Pathogenicity tests with mice for all the isolates confirmed their identity by the demonstration of large yeast forms (8-15 microns in diameter) within giant cells in the infected tissues. Investigations on the possible occurrence of human infections in the area are in progress.

Animals↗

A new natural reservoir of hantavirus: isolation of hantaviruses from lung tissues of bats.

Two species of bats were confirmed as new natural reservoirs of hantavirus. Antibodies to Hantaan virus were detected in 3.40% (23 of 677) of bats captured from 1989 to 1992 in Korea by the IFA technique. Areal distribution of immunofluorescent antibody were different, and seropositive rates were much high in sera of bats captured in summer (3.82%) and winter (5.82%). Viral antigens were observed in the lungs (3 of 16) and kidney (1 of 7). Two hantaviruses were isolated from lung tissues of E. serotinus and R. ferrum-equinum through a cell culture system, designated CUMC-92B8 and -92B48, respectively. Using Rous associated virus-2 reverse transcriptase-directed PCR and 2 oligonucleotide primer pairs, genomic sequences of the isolates were amplified. Amplified products of the isolates and reactivities to monoclonal antibodies very closely resembled those of Hantaan virus. These data suggest that the serotype of the isolates is closely related to Hantaan virus, and bats serve as reservoirs of hantavirus.

Animals↗