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Cochlear hair-cell populations and limits of resolution of hearing in two vespertilionid bats, Nyctalus noctula and Eptesicus serotinus.

A comparison of the population of cochlear hair cells with known hearing resolution capabilities in two species of bats (Nyctalus noctula and Eptesicus serotinus of the family Vespertilionidae) has revealed a close correlation between morphological and functional parameters. Data on number and densities of hair cells in the examined bat species are presented. Even though the species are of a comparable body size, N. noctula has a longer basilar membrane, a higher density, and hence also a higher total number of hair cells than E. serotinus. This is in good agreement with hearing resolution capabilities, which are better in N. noctula than in E. serotinus. These findings have implications on differences in feeding ecology and hunting strategies between these species.

Animals↗

Ultrastructural changes in the seminiferous epithelium of two seasonally reproducing bats (Mammalia: Chiroptera).

The Sertoli cells of the Cape horseshoe bat (Rhinolophus capensis) and Schreiber's long-fingered bat (Miniopterus schreibersii) undergo marked changes in ultrastructure related to stages in the spermatogenic cycle. The amount of lipid stored in the Sertoli cells varies annually and is at a maximum from just after spermiation to early in the following spermatogenic cycle. During spermatogenesis, the diameter of the lipid droplets decreases, reaching a minimum prior to spermiation. Sertoli cells exhibit a marked apicobasal differentiation, particularly in the vicinity of developing late spermatids, where the cytoplasm of the Sertoli cell is packed with smooth endoplasmic reticulum. The possible roles of lipid droplets and smooth endoplasmic reticulum. The possible roles of lipid droplets and smooth endoplasmic reticulum in steroidogenesis by Sertoli cells are discussed. Junctional complexes occur between Sertoli cells and spermatogonia, are apparently absent from between Sertoli cells and spermatocytes, and are restricted to the region of the developing acrosome in the spermatids. Annulate lamellae, which occur commonly in the developing germinal cells and less frequently in the Sertoli cells, may be associated with the production of microtubules, which are present in both spermatids and Sertoli cells.

Animals↗

Adrenergic innervation of the heart of the bat, Miniopterus schreibersii.

The adrenergic innervation of the heart of the bat (Miniopterus schreibersii) was studied with a fluorescence histochemical technique. The appearance and distribution pattern of the terminal adrenergic nerve fibers demonstrated in the atria, sinoatrial and atrioventricular nodes, and ventricles is typically mammalian. Fine varicose adrenergic fibers run in and parallel to the ventricular muscle where they are common and uniformly distributed. Indian ink perfusion of the coronary vasculature demonstrates the high density of vessels in the ventricles, but obscures the terminal innervation of the ventricular muscle. This alone, or in combination with the apparent seasonal change in the terminal innervation of the ventricles, may explain the inability of previous workers to identify an adrenergic ventricular innervation in the bat Nyctalus noctula. The adrenergic ventricular innervation might be involved in the large increases in cardiac output associated with the commencement of flight and in the massive sympathetic activation that mediates arousal from torpor.

Animals↗

Histochemical and myosin composition of vampire bat (Desmodus rotundus) pectoralis muscle targets a unique locomotory niche.

The vampire bat pectoralis muscle contains at least four fiber types distributed in a nonhomogeneous pattern. One of these fiber types, here termed IIe, can be elucidated only by adenosine triphosphatase (ATPase) histochemistry combined with reactions against antifast and antislow myosin antibodies. The histochemical and immunohistochemical observations indicate a well-developed specialization of function within specific regions of the muscle. In parallel, analyses of native myosin isoforms and myosin heavy chain isoforms indicate two points. First, the histochemical "type IIe" fiber is predominant in cranial portions of the muscle, and myosin extracted from these regions exhibits a unique electrophoretic mobility not observed in the myosin isoforms of more traditional laboratory mammals. Second, the type I fibers are confined to the pectoralis abdominalis muscle and a small adjacent region of the caudal part of the pectoralis. This pattern of type I fiber distribution is considered a derived character state compared to muscle histochemical phenotype and isoform composition in the pectoralis muscles of other phyllostomids we have studied (Artibeus jamaicensis, Artibeus lituratus, Carollia perspicillata). We relate this to the unique locomotory needs of the common vampire bat, Desmodus rotundus.

Animals↗

Pectoralis muscle morphology in the little brown bat, Myotis lucifugus: a non-convergence with birds.

Recent studies of muscle architecture demonstrate that many mammalian muscles are composed of short, interdigitating fibers. In addition, the avian pectoralis, a muscle capable of producing high frequency oscillations has been shown to possess a serially arranged pattern of muscle endplate in all sizes of birds studied. The pectoralis muscle of the little brown bat, Myotis lucifugus (Chiroptera: Vespertilionidae), is composed of fairly uniform fibers that span the length of the muscle and is characterized by a zone of motor endplates within the middle third of the muscle. The homogeneous fiber architecture of the bat pectoralis muscle is in contrast to the serial arrangement of endplates (and presumably muscle fibers) in the avian pectoralis in species equivalent in size to Myotis. The short fiber organization and motor endplate pattern observed in most birds is thus not a requisite design for flying vertebrates.

Animals↗

Ontogeny of the pectoralis muscle in the little brown bat, Myotis lucifugus.

The ontogeny of a primary flight muscle, the pectoralis, in the little brown bat (Myotis lucifugus: Vespertilionidae) was studied using histochemical, immunocytochemical, and electrophoretic techniques. In fetal and early neonatal (postnatal age 1-6 days) Myotis, histochemical techniques for myofibrillar ATPase (mATPase) and antibodies for slow and fast myosins demonstrated the presence of two fiber types, here called types I and IIa. These data correlated with multiple transitional myosin heavy chain isoforms and native myosin isoforms demonstrated with SDS-PAGE and 4% pyrophosphate PAGE. There was a decrease in the distribution and number of type I fibers with increasing postnatal age. At postnatal age 8-9 days, the adult phenotype was observed with regard to muscle fiber type (100% type IIa fibers) and myosin isoform profile (single adult MHC and native myosin isoforms). This "adult" fiber type profile and myosin isoform composition preceded adult function by about 2 weeks. For example, little brown bats were incapable of sustained flight until approximately postnatal day 24, and myofiber size did not achieve adult size until approximately postnatal day 25. Although Myotis pectoralis is unique in being composed of 100% type IIa fibers, transitional fiber types and isoforms were present. These transitional forms had been observed previously in other mammals bearing mixed adult muscle fibers and which undergo transitional stages in muscle ontogeny. However, in Myotis pectoralis, this transition transpires relatively early in development.

Aging↗

Protein kinase A from bat skeletal muscle: a kinetic study of the enzyme from a hibernating mammal.

The catalytic subunit of adenosine 3'-5'-cyclic mono-phosphate-dependent protein kinase (PKAc) was purified to homogeneity from skeletal muscle of the little brown bat, Myotis lucifugus. The purification procedure was highly reproducible, resulting in a final activity of 205 nmol phosphate transferred/min/mg protein at 22 degreesC. Identification of the enzyme as a protein kinase A was confirmed through the use of specific PKA inhibitors. The catalytic subunit had a molecular weight of 54.6 +/- 3.5 kDa.Km values for Kemptide and Mg-ATP were 9.1 +/- 0.2 and 94.1 +/- 4.5 microM at 37 degreesC, respectively. Both values decreased significantly at 5 degreesC to 37 and 52% of their values at the higher temperature. Similar temperature effects on Km values were found with the purified commercial pig heart enzyme. Neutral salts had little effect on enzyme activity (I 50 values >400 mM) but NaF had an I 50 of 38 mM; except for fluoride, ions were less inhibitory at 5 degreesC, compared with 37 degreesC. Arrhenius plots showed evidence of a temperature-dependent conformational change; a distinct break in the plot occurred at 10 degreesC giving calculated activation energies of 5.6 +/- 0. 46 kJ/ mol at temperatures above 10 degreesC++and 29.5 +/- 2.0 kJ/mol below 10 degreesC. Porcine PKAc, by contrast, showed a linear Arrhenius plot over the entire temperature range tested and an intermediate activation energy of 15.9 +/- 0.3 kJ/mol. The pH optimum of bat PKAc also changed dramatically with temperature falling from 8.5 at 37 degreesC to 5.5 at 5 degreesC, an effect that could substantially change enzyme activity in vivo at the low body temperature of the hibernating state. Overall, low temperature had both positive (increased the percentage of PKAc, reduced Km values, increased I 50 values for salts) and negative (increased activation energy, acidic shift of pH optimum) effects on PKAc but the substantial positive effects of low temperature on the enzyme suggest an important role for continued PKA action in signal transduction in the hibernating animal.

Animals↗

Effects of gonadal sex steroids on sexual behavior in the big brown bat, Eptesicus fuscus, upon arousal from hibernation.

Vespertilionid bats are thought to have a dissociated pattern of reproduction; mating occurs in autumn as well as during periodic arousals from hibernation when testes are regressed and ovaries are "in stasis." Sex steroid levels in both sexes are basal at this time. This pattern would indicate that sex steroids per se may not be activating sexual behavior. Gonadectomy of male and female big brown bats, Eptesicus fuscus, months before the mating season, did not significantly affect the probability of expressing sexual behavior in either sex when staged behavior tests were conducted. Ovariectomized females, in fact, mated significantly more and exhibited proceptive behavior. Males mated at the same frequency whether they were intact, orchidectomized, or given testosterone implants. These results argue that gonadal sex steroids are not the direct activators of sexual behavior in this species. However, when males were allowed to mate freely within a large flight cage, orchidectomized males were not seen to mate, while intact males mated at the same frequency as in the staged trials. Females are apparently either differentiating between the two groups of males or there is male-male competition for mates within a system that has multiple matings with different individuals in both sexes.

Animals↗

Molecular systematics of bats of the genus Myotis (Vespertilionidae) suggests deterministic ecomorphological convergences.

Based on extensive phenetic analyses, bats of the genus Myotis have been classically subdivided into four major subgenera each of which comprise many species with similar morphological and ecological adaptations. Each subgenus thus corresponds to a distinct "ecomorph" encompassing bat species exploiting their environment in a similar fashion. As three of these subgenera are cosmopolitan, regional species assemblages of Myotis usually include sympatric representatives of each ecomorph. If species within these ecomorphs are monophyletic, such assemblages would suggest extensive secondary dispersal across geographic areas. Conversely, these ecomorphological adaptations may have evolved independently through deterministic processes, such as adaptive radiation. In this case, phylogenetic reconstructions are not expected to sort species of the same ecomorph into monophyletic clades. To test these predictions, we reconstructed the phylogenetic history of 13 American, 11 Palaearctic, and 6 other Myotis species, using sequence data obtained from nearly 2 kb of mitochondrial genes (cytochrome b and nd1). Separate or combined analyses of these sequences clearly demonstrate the existence of several pairs of morphologically very similar species (i.e., sibling species) which are phylogenetically not closely related. None of the three tested subgenera constitute monophyletic units. For instance, Nearctic and Neotropical species currently classified into the three subgenera were clustered in a single, well-supported monophyletic clade. These species thus evolved independently of their ecological equivalents from the Palaearctic region. Independent adaptive radiations among species of the genus Myotis therefore produced strikingly similar evolutionary solutions in different parts of the world. Furthermore, all phylogenetic reconstructions based on mtDNA strongly supported the existence of an unsuspected monophyletic clade which included all assayed New World species plus M. brandtii (from the Palaearctic Region). This "American" clade thus radiated into a morphologically diverse species assemblage which evolved after the first Myotis species colonized the Americas. Molecular reconstructions support paleontological evidence that species of the genus Myotis had a burst of diversification during the late Miocene-early Pliocene epoch.

Animals↗

Sequence analysis of an isolate from a fatal human infection of Australian bat lyssavirus.

Australian bat lyssavirus (ABLV), which occurs in pteropid and insectivorous bat populations, causes a rabies-like encephalitis in infected humans. We report the first complete sequence of an ABLV isolate obtained from a human who developed symptoms 27 months after being bitten by an infected flying fox. This isolate is the smallest lyssavirus to be sequenced, with a size of 11,918 nucleotides. Analyses of previously unsequenced regions and the complete genome confirm its close relationship with classical rabies viruses. In addition, a leucine zipper-like motif, not present in the other lyssaviruses, was found in the conserved domain I of the polymerase protein. This is the first report of a lyssavirus to vary in an 11-nucleotide, strictly conserved, complementary terminal sequence. This region is thought to encode important cis-acting regulatory signals; ABLV variation indicates a greater degree of flexibility than was thought for lyssaviruses in this region. A comparison of the pteropid and insectivorous isolates of ABLV indicates considerable differences between the two viruses. If the divergence of the two occurred on the Australian mainland, ABLV may have been endemic to Australia well before European colonisation.

Adult↗

Novel viral encephalitides associated with bats (Chiroptera)--host management strategies.

Several novel viruses recently described in bats of the genus Pteropus (sub-order Megachiroptera) in Australia and southeast Asia cause encephalitic disease in animals and humans. These viruses include Hendra virus and Nipah virus (genus Henipavirus, family Paramyxoviridae) and Australian bat lyssavirus (ABLV; genus Lyssavirus, family Rhabdoviridae). Broadly, strategies for disease prevention and control in the spillover host are directed at minimising direct or indirect contact with the natural host, improving farm-gate and on-farm biosecurity, and better disease recognition and diagnosis. Additional strategies for ABLV include the use of rabies vaccine for effective pre- and post-exposure prophylaxis in humans. Effective management strategies in the natural host are predicated on an understanding of the ecology of the disease in the natural host, and the identification and avoidance of factors putatively associated with emergence, such as habitat loss, land use change and demographic shifts. A possible future management strategy for ABLV in reservoir populations is immunisation using bait or plant-derived vaccination.

Animals↗

Spatial processing within the mustache bat echolocation system: possible mechanisms for optimization.

1. The directionality of an echolocation system is determined by the acoustic properties of both the emitter and receiver, i.e., by the radiation pattern of the emitted pulse and the directionally of the external ears. We measured the directionality of the echolocation system of the greater mustache bat (Pteronotus parnellii) at the 30 kHz, 60 kHz and 90 kHz harmonics of its echolocation pulse by summing, at points throughout the frontal sound field, the echo attenuation due to the spread of pulse energy and the attenuation due to the spread of pulse energy and the attenuation due to the directionality of its external ears. The pulse radiation pattern at the 3 harmonics was measured by comparing the output of a microphone moved throughout the frontal sound field against a second reference microphone at the center of the field. External ear directionality at the 3. harmonics was measured by presenting free-field sounds throughout the frontal sound field, and recording the intensity thresholds of cochlear microphonic potentials, and the intensity thresholds of monaural neurons in the inferior colliculus tuned to one of the 3 harmonics. 2. When compared with ear directionality, the echolocation system was found to be more directional for the center of the sound field in several respects. At all harmonics, attenuation of sounds originating in the peripheral part of the field was increased by 10 to 13 dB. Areas of maximum sound intensity contracted toward the center of the field. Also, the isointensity contours of the echolocation system were more radially symmetrical about the center of the field. 3. At 60 kHz, sound intensity along the azimuth within the echolocation system was nearly constant 26 degrees to either side of the center of the field. This suggests that the radiation pattern of the echolocation pulse and the directionality of the external ears complement one another to produce an acoustic environment at the center of the sound field in which stimulus intensity is stabilized to allow more effective analysis of various aspects of the echolocation target. In particular, we suggest that this intensity stabilization may allow the bat to more effectively resolve the interaural intensity differences it uses to localize prey. 4. Predictions of the azimuthal spatial tuning of binaurally sensitive neurons in the inferior colliculus within the echolocation system were compared with their spatial tuning when only ear directionality is considered.(ABSTRACT TRUNCATED AT 400 WORDS)

Acoustic Stimulation↗

The effect of pulse repetition rate on the delay sensitivity of neurons in the auditory cortex of the FM bat, Myotis lucifugus.

1. Echo delay is the primary cue used by echolocating bats to determine target range. During target-directed flight, the repetition rate of pulse emission increases systematically as range decreases. Thus, we examined the delay tuning of 120 neurons in the auditory cortex of the bat, Myotis lucifugus, as repetition rate was varied. 2. Delay sensitivity was exhibited in 77% of the neurons over different ranges of pulse repetition rates (PRRs). Delay tuning typically narrowed and eventually disappeared at higher PRRs. 3. Two major types of delay-sensitive neurons were found: i) delay-tuned neurons (59%) had a single fixed best delay, while ii) tracking neurons (22%) changed their best delay with PRR. 4. PRRs from 1-100/s were represented by the population of delay-sensitive neurons, with the majority of neurons delay-sensitive at PRRs of at least 10-20/s. Thus, delay-dependent neurons in Myotis are most active during the search phase of echolocation. 5. Delay-sensitive neurons that also responded to single sounds were common. At PRRs where delay sensitivity was found, the responses to single sounds were reduced and the responses to pulse-echo pairs at particular delays were greater than the single-sound responses. In facilitated neurons (53%), the maximal delay-dependent response was always larger than the best single-sound responses, whereas in enhanced neurons (47%), these responses were comparable. The presence of neurons that respond maximally to single sounds at one PRR and to pulse-echo pairs with particular echo delays at other PRRs suggests that these neurons perform echo-ranging in conjunction with other biosonar functions during target pursuit.

Acoustic Stimulation↗

Spectral selectivity of FM-FM neurons in the auditory cortex of the echolocating bat, Myotis lucifugus.

1. Spectral sensitivity was examined in delay-sensitive neurons in the auditory cortex of the awake FM bat, Myotis lucifugus. FM stimuli sweeping 60 kHz downward in 4 ms were used as simulated pulse-echo pairs to measure delay-dependent responses. At each neuron's best delay, the pulse and/or echo were divided into 4 FM quarters (Ist, IInd, IIIrd, and IVth), each sweeping 15 kHz in 1 ms, and quarters essential for delay sensitivity were determined for both pulse and echo. 2. For the pulse, the IVth quarter was essential for delay sensitivity in the majority of neurons. For the echo, the essential quarter for most neurons was the IInd, IIIrd, or IVth. 3. Different quarters of the pulse and echo were essential for delay sensitivity in 68% of the neurons examined. 4. This study provides neurophysiological evidence linking both spectral and temporal processing in delay-sensitive neurons of Myotis. Since spectral cues can provide target-shape information, sensitivity to both spectral and temporal parameters in single neurons may endow these neurons in FM bats with the potential for target analysis other than echo-ranging.

Acoustic Stimulation↗

Using artificial bat sonar neural networks for complex pattern recognition: recognizing faces and the speed of a moving target.

Two sets of studies examined the viability of using bat-like sonar input for artificial neural networks in complex pattern recognition tasks. In the first set of studies, a sonar neural network was required to perform two face recognition tasks. In the first task, the network was trained to recognize different faces regardless of facial expressions. Following training, the network was tested on its ability to generalize and correctly recognize faces using echoes of novel facial expressions that were not included in the training set. The neural network was able to recognize novel echoes of faces almost perfectly (above 96% accuracy) when it was required to recognize up to five faces. In the second face recognition task, a sonar neural network was trained to recognize the sex of 16 faces (eight males and eight females). After training, the network was able to correctly recognize novel echoes of those faces as 'male' or as 'female' faces with accuracy levels of 88%. However, the network was not able to recognize novel faces as 'male' or 'female' faces. In the second set of studies, a sonar neural network was required to learn to recognize the speed of a target that was moving towards the viewer. During training, the target was presented in a variety of orientations, and the network's performance was evaluated when the target was presented in novel orientations that were not included in the training set. The different orientations dramatically affected the amplitude and the frequency composition of the echoes. The neural network was able to learn and recognize the speed of a moving target, and to generalize to new orientations of the target. However, the network was not able to generalize to new speeds that were not included in the training set. The potential and limitations of using bat-like sonar as input for artifical neural networks are discussed.

Adult↗

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

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 2). 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 interference between 400 and 550 Hz (Fig. 1). 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 (Fig. 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 (Fig. 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).(ABSTRACT TRUNCATED AT 250 WORDS)

Acoustic Stimulation↗

Organochlorine pesticide accumulation and genotoxicity in Mexican free-tailed bats from Oklahoma and New Mexico.

The summer population of Mexican free-tailed bats (Tadarida brasiliensis) at Carlsbad Caverns, New Mexico, declined from an estimated 8.7 million in 1936 to 200,000 in 1974; thereafter, it increased to approximately 700,000 in 1991. This decline has been attributed primarily to organochlorine (OC) pesticide contamination and habitat disturbance. Similar declines have been observed in other populations of this species. This study examined the potential genotoxic effects of OC pesticide contamination on two populations of T. brasiliensis. Pesticide accumulation, frequencies of chromosomal aberrancy, and nuclear DNA content variation in spleen and testicular tissues were examined in specimens collected from Carlsbad Caverns and Vickery Cave, a maternity colony in northwestern Oklahoma, during the summers of 1990 and 1991. Pesticide residues in brain and carcass tissues were identified and quantified by electron capture gas chromatography. Genotoxicity was examined with the standard bone marrow chromosomal aberration assay and flow cytometry. Statistical relationships among pesticide content, observed chromosomal aberrancy, and nuclear DNA content variation were examined. Both populations demonstrated significant levels of DDE contamination; however, the Carlsbad Caverns population showed consistently higher pesticide loads. Males also demonstrated higher levels than females. No statistical differences in chromosomal aberrancy or nuclear DNA content variation were observed among sexes, sites, or collected periods. Positive correlations were detected between brain and carcass DDE concentrations for all bats examined. A significant negative relationship was found between brain DDE concentration and coefficients of variation in spleen DNA content only for males.

Analysis of Variance↗

Frequency and space representation in the inferior colliculus of the FM bat, Eptesicus fuscus.

The tonotopic organization and spatial sensitivity of 217 inferior collicular (IC) neurons of Eptesicus fuscus were studied under free field stimulation conditions. Acoustic stimuli were delivered from a loudspeaker placed 21 cm ahead of the bat to determine the best frequency (BF) and minimum threshold (MT) of isolated IC neurons. A BF stimulus was then delivered as the loudspeaker was moved horizontally across the frontal auditory space of the bat to locate the best azimuthal angle (BAZ) at which the neuron had its lowest MT. The stimulus was then raised 3 dB above the lowest MT to determine the horizontal extent of the auditory space within which a sound could elicit responses from the neurons. This was done by moving the loudspeaker laterally at every 5 degrees or 10 degrees until the neuron failed to respond. These measurements also allowed us to redetermine the BAZ at which the neuron fired maximal number of impulses. Electrodes were placed evenly across the whole IC surface and IC neurons were sampled as many as possible within each electrode penetration. Tonotopic organization and spatial sensitivity were examined among all 217 IC neurons as a whole as well as among IC neurons sequentially sampled within individual electrode penetrations. The whole population of 217 IC neurons is organized tonotopically along the dorsoventral axis of the IC. Thus, low frequency neurons are mostly located dorsally and high frequency neurons ventrally with median frequency neurons intervening in between. The BAZ of these 217 IC neurons tend to shift from lateral to medial portions of the contralateral frontal auditory space with increasing BF.(ABSTRACT TRUNCATED AT 250 WORDS)

Acoustic Stimulation↗