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Evaluation of DNA damage in a population of bats (Chiroptera) residing in an abandoned monazite mine.

Ionising radiation has the ability to induce DNA damage. While the effects of high doses of radiation of short duration have been well documented, the biological effects of long-term exposure to low doses are poorly understood. This study evaluated the clastogenic effects of low dose ionising radiation on a population of bats (Chiroptera) residing in an abandoned monazite mine. Bats were sampled from two chambers in the mine, where external radiation levels measured around 20 microSv/h (low dose) and 100 microSv/h (higher dose), respectively. A control group of bats was sampled from a cave with no detectable radiation above normal background levels. The micronucleus assay was used to evaluate residual radiation damage in binucleated lymphocytes and showed that the micronucleus frequency per 500 binucleated lymphocytes was increased in the lower radiation-exposed group (17.7) and the higher radiation-exposed group (27.1) compared to the control group (5.3). This study also showed that bats exposed to radiation presented with an increased number of micronuclei per one thousand reticulocytes (2.88 and 10.75 in the lower and high radiation-exposed groups respectively) when compared to the control group (1.7). The single-cell gel electrophoresis (comet) assay was used as a means of evaluating clastogenecity of exposure to radiation at the level of individual cells. Bats exposed to radiation demonstrated increased DNA damage as shown by the length of the comet tails and showed an increase in cumulative damage. The results of the micronucleus and the comet assays indicated not only a statistically significant difference between test and control groups (P<0.001), but also a dose-dependent increase in DNA damage (P<0.001). These assays may thus be useful in evaluating the potential clastogenecity of exposure to continuous low doses of ionising radiation.

Animals↗

Alterations in surface activity of pulmonary surfactant in Gould's wattled bat during rapid arousal from torpor.

The small microchiropteran bat, Chalinolobus gouldii, undergoes large daily fluctuations in metabolic rate, body temperature, and breathing pattern. These alterations are accompanied by changes in surfactant composition, predominantly an increase in cholesterol relative to phospholipid during torpor. Furthermore, the surface activity changes, such that the surfactant functions most effectively at that temperature which matches the animal's activity state. Here, we examine the surface activity of surfactant from bats during arousal from torpor. Bats were housed at 24 degrees C on an 8:16h light:dark cycle and their surfactant was collected during arousal (28<T(b)<32 degrees C). Surface activity was examined with a Captive Bubble Surfactometer at 24 and 37 degrees C. Surfactant from arousing bats was more active at 37 degrees C than at 24 degrees C, indicated by a lower ST(min) and reduced film area compression required to reach ST(min). It appears that the arousal-induced changes in surfactant composition, i.e., lower levels of cholesterol, inhibit adsorption of surfactant at 24 degrees C. Furthermore, the alterations in surfactant composition during arousal are very rapid, such that the mixture behaves more like surfactant from warm-active bats, and therefore, functions more effectively at 37 degrees C.

Adaptation, Physiological↗

Role of immunoglobulin classes in experimental histoplasmosis in bats.

Pooled normal bat serum was separated by gel filtration to give fractions rich in IgG-, Iga- and IgM-like proteins. These fractions were analogous to the corresponding human immunoglobulin classes by immunoelectrophoresis and SDS-polyacrylamide gel electrophoresis. Rabbits were immunized with the fractions and the antisera absorbed. Neotropical bats (Artibeus lituratus) were infected with Histoplasma capsulatum and serum samples were collected weekly and tested for specific serologic response to the fungus. A radial immunodiffusion test was devised to monitor changes in concentrations of IgG, IgA and IgM in the same sera. Bats infected with a low dose of fungus had significantly increased levels of IgM and IgA between 2-6 weeks post-infection. Bats receiving a high dose maintained elevated levels of IgM and IgA through the end of the study. Significantly elevated levels of IgG were not detected until late in the disease (8-9 weeks). In bats with histoplasmosis, IgM and IgA appeared to contribute primarily to the early positive serologies, while precipitating antibodies of the IgG class were detectable later in the disease. These results are similar to the serologic profile seen in human histoplasmosis, and extend our understanding of comparative immune responses in an important wildlife reservoir of human mycotic pathogens.

Animals↗

Antigenic and genetic divergence of rabies viruses from bat species indigenous to Canada.

Antigenic characterisation of over 350 chiropteran rabies viruses of the Americas, especially from species reported rabid in Canada, distinguished 13 viral types. In close accord with this classification, nucleotide sequencing of representative isolates, at both the N and G loci, identified four principal phylogenetic groups (I-IV), sub-groups of which circulated in particular bat species. Amongst the North American bat viruses, there was a notable division between group I specimens associated with colonial, non-migratory bats (Myotis sp. and Eptesicus fuscus) and those of group II harbored by solitary, migratory species (Lasiurus sp. and Lasionycteris noctivagans). Certain species of Myotis were clearly identified as rabies reservoirs, an observation often obscured previously by their frequent infection by viral variants of other chiroptera. An additional group (III) apparently circulates in E. fuscus, whilst viruses harbored by both insectivorous and haematophagus bats of Latin America clustered to a separate clade (group IV). Comparison of the predicted N and G proteins of these viruses with those of strains of terrestrial mammals indicated a similarity in structural organisation regardless of host species lifestyle. Finally, these sequences permitted examination of the evolutionary relationship of American bat rabies viruses within the Lyssavirus genus.

Animals↗

Sound-evoked efferent effects on cochlear mechanics of the mustached bat.

The influence of the crossed medial efferent system on cochlear mechanics of the mustached bat was tested by measuring delayed evoked otoacoustic emissions (DEOAEs), cochlear microphonics, distortion product otoacoustic emissions (DPOAEs) and stimulus frequency otoacoustic emissions. Contralaterally delivered sinusoids, broadband noise and bat echolocation calls were used for acoustic stimulation of the efferent system. With all four measures we found a level-dependent suppression under stimulation with both broadband noise and echolocation calls. In addition, the sharply tuned cochlear resonance of the mustached bat which is involved in processing echolocation signals at 61 kHz shifted upward in frequency by several 100 Hz. Presentation of sinusoids did not have any significant effect. DEOAEs and DPOAEs were in some cases enhanced during contralateral presentation of the bat calls at moderate intensities. The most important function of the efferent system in the mustached bat might be the control of the extraordinarily fine-tuned resonator of this species, which is close to instability as evident from the very pronounced evoked otoacoustic emissions which sometimes convert into spontaneous otoacoustic emissions of high level.

Acoustic Stimulation↗

Passive sound-localization ability of the big brown bat (Eptesicus fuscus).

The passive sound-localization ability (i.e. minimum audible angle) of the big brown bat, Eptesicus fuscus, was determined using a conditioned avoidance procedure in which the animals were trained to discriminate left sounds from right sounds. The mean threshold of three bats for a 100-ms broadband noise burst was 14 degrees, a value that is about average for mammals. A similar threshold of 15 degrees was obtained for one animal when it was retested with one of its own recorded echolocation calls as the stimulus. The two bats tested on pure-tone localization were able to localize high-frequency, but not low-frequency tones, even when a low-frequency tone was amplitude modulated, a result indicating that these bats are not able to use binaural time-difference cues for localization. Finally, given the width of the bat's field of best vision, as determined by a count of its ganglion-cell density, its sound-localization acuity is consistent with the hypothesis that the role of passive sound localization is to direct the eyes to the source of a sound.

Acoustic Stimulation↗

Rapid GLUT-1 mediated glucose transport in erythrocytes from the grey-headed fruit bat (Pteropus poliocephalus).

D-Glucose entry into erythrocytes from adult grey-headed flying fox fruit bats (Pteropus poliocephalus) was rapid and showed saturation at high substrate concentrations. Kinetic parameters were estimated from the concentration dependence of initial rates of zero-trans D-glucose entry at 5.5 degrees C as Michaelis constant (K(m)) 1. 64+/-0.56 mM, and maximal velocity (V(max)) 1162+/-152 micromol.l. cell water(-1).min(-1). D-Glucose entry was inhibited by cytochalasin B; mass law analysis of D-glucose-displaceable cytochalasin B binding gave values of K(d) 37.1+/-5.0 nM and B(max) 361.2+/-9.1 pmol/mg membrane protein. Entry of 2-deoxy-D-glucose, and 3-O-methyl-D-glucose, into P. poliocephalus red cells was rapid, entry of D-fructose was very slow. Glucose transporter polypeptides were identified on immunoblots as a band M(r) 47000-54000 and their identity confirmed by D-glucose-sensitive photolabeling of membranes with [3H]-cytochalasin B. Peptide-N-glycanase F digestion of both human and bat erythrocyte membranes generated GLUT-1-derived bands M(r) 37000. Trypsin digestion of human and fruit bat erythrocyte membranes generated fragmentation patterns consistent with similar GLUT-1 polypeptide structures in both species. Erythrocytes from adult Australian ghost bats (Macroderma gigas), a carnivorous microchiropteran bat, also expressed high levels of GLUT-1.

Amidohydrolases↗

A new species of trypanosome, Trypanosoma desterrensis sp. n., isolated from South American bats.

Trypanosomes isolated from South American bats include the human pathogen Trypanosoma cruzi. Other Trypanosoma spp. that have been found exclusively in bats are not well characterized at the DNA sequence level and we have therefore used the SL RNA gene to differentiate and characterize kinetoplastids isolated from bats in South America. A Trypanosoma sp. isolated from hats in southern Brazil was compared with the geographically diverse isolates T. cruzi marinkellei, T. vespertilionis, and T. dionisii. Analysis of the SL RNA gene repeats revealed size and sequence variability among these bat trypanosomes. We have developed hybridization probes to separate these bat isolates and have analysed the DNA sequence data to estimate their relatedness. A new species, Trypanosoma desterrensis sp. n., is proposed, for which a 5S rRNA gene was also found within the SL RNA repeat.

Animals↗

Molecular characterization of haemoparasites infecting bats (Microchiroptera) in Cornwall, UK.

The presence of haemoparasites from the Order Piroplasmida and the genera Bartonella and Trypanosoma was assessed in the blood of 60 bats, belonging to 7 species, inhabiting sites across Cornwall in southwest England. DNA extracted from macerated heart tissue was incorporated into taxon-specific polymerase chain reactions (PCRs) and amplification products were sequenced as a means of identifying, or assigning an identity, to detected haemoparasites. A Piroplasmida species was detected in 6 Pipistrellus spp., whereas Bartonella infections were detected in 5 bats belonging to 4 different species. Trypanosoma dionisii was detected in 1 Pipistrellus spp. Phylogenetic inference from alignment of a partial 18S rRNA-encoding gene sequence of the pipistrelle-associated Piroplasmida species with homologous sequences available for other members of the Order indicated that this organism was unique but specifically related to members of the genus Babesia, a phylogeny that would be in keeping with the organism being Babesia vesperuginis. Alignment of partial citrate synthase gene sequences from the bat-associated bartonellae revealed 5 distinct genotypes that were probably derived from 2 distinct Bartonella species. The study demonstrates the utility of molecular methods for detecting haemoparasites in dead bats and provides, for the first time, tangible identities for bat-associated Babesia and Bartonella species.

Animals↗

Hearing in a megachiropteran fruit bat (Rousettus aegyptiacus).

The Egyptian fruit bat (Rousettus aegyptiacus) is one of the few megachiropteran bats capable of echolocation. However, it uses rudimentary tongue clicks rather than laryngeally produced echo calls. We determined the audiogram of 2 bats using a conditioned avoidance procedure with fruit puree reward. At an intensity of 60 dB sound pressure level, the bats' hearing extended from 2.25 kHz to 64 kHz, with a region of good sensitivity between 8 kHz and 45 kHz. A dip in sensitivity at 32 kHz appears to be due to pinna directionality. The hearing of Egyptian fruit bats is typical for a mammal of that size and is not as limited as previously reported. Methodological issues, specifically training an animal to listen for low-intensity signals and imposing a significant cost for failing to report signals (i.e., misses), are discussed as the basis for the discrepancy between our results and earlier reports.

Animals↗

Sound localization in an Old-World fruit bat (Rousettus aegyptiacus): acuity, use of binaural cues, and relationship to vision.

The passive sound-localization acuity of Egyptian fruit bats (Rousettus aegyptiacus) was determined using a conditioned-avoidance procedure. The mean minimum audible angle for left-right discrimination for 3 bats was 11.6 degrees--very near the mean for terrestrial mammals. The bats also were able to localize low- and high-frequency pure tones, indicating that they can use both binaural phase-difference and binaural intensity-difference cues to localize sound. Moreover, they were able to use the binaural phase-difference cue up to at least 5.6 kHz, which is higher than other mammals yet tested. The width of the Egyptian fruit bats' field of best vision was 27 degrees. This value is consistent with the hypothesis that the role of passive sound localization is to direct the eyes for visual scrutiny of sound sources. Thus, the passive localization abilities of these echolocating megachiropteran fruit bats do not deviate from the patterns established for nonecholocating mammals.

Animals↗

A possible neuronal basis for Doppler-shift compensation in echo-locating horseshoe bats.

The auditory system of the horseshoe bat is finely tuned to the bat's individual vocalization frequency. To compensate for flight-induced Doppler shifts in the echo frequency, the horseshoe bat adjusts the frequency of its echo-location call to maintain the echo frequency within the narrow range to which its auditory system is best tuned. In this report I describe neurons in the midbrain tegmentum of the horseshoe bat, with properties that strongly indicate their involvement in this Doppler-shift compensation. The activity of these neurons was influenced by both sound emission and auditory stimuli. Neuronal discharges in response to vocalization, however, differed from those in response to purely auditory stimuli that mimicked the bat call. When an auditory stimulus was temporally locked to a preceding vocalization, the response was dependent on the time delay between the two. This delay-sensitivity completely disappeared when vocalizations were simulated acoustically. Only those vocalization and 'echo' parameters were encoded that occur in Doppler-shift compensation. In conclusion, I suggest a model for the regulation of the vocalization frequency through auditory feedback.

Animals↗

Harmonic-hopping in Wallacea's bats.

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

Acoustics↗

An adjustable table tennis bat and grip system for tetraplegics.

A table tennis bat and gripping system has been devised which can be used by people who are tetraplegic who are unable to hold the bat unassisted. The multi-positional head of the bat allows the user to alter the bat angle with respect to the hand whilst an elasticated glove ensures the bat is held firmly in place allowing the player access to a greater range of playing shots. The system has been used successfully to introduce recently injured players to the game, and has also been used by a British ranked tetraplegic table tennis player. It is suitable for general, rehabilitative or competition use.

Hand↗

Seasonal reproduction and host infestation rates for nycteribiids of the large bentwing bat.

Large numbers of the nycteribiid species Nycteribia parilis vicaria Maa, Penicillidia (Penicillidia) oceanica Bigot and Penicillidia (Penicillidia) Maa (Diptera: Nycteribiidae) were collected from the bat Miniopterus schreibersii Kuhl (Chiroptera: Vespertilionidae) during a study on this bat's population genetics conducted largely in southern Australia. Bats were trapped in all seasons and at a number of locations throughout the ranges of the nycteribiids. Nycteribia parilis vicaria was the most common species captured. Gravid females of the three species were trapped in spring and summer and gravid female P. (P.) oceanica and N. parilis vicaria were trapped in autumn. Newly emerged adults of P. (P.) oceanica and N. parilis vicaria were captured in spring and autumn. Nycteribiid loads for individual bats were low, and there was no direct evidence of a detrimental effect on bats.

Animals↗

The central acoustic tract and audio-vocal coupling in the horseshoe bat, Rhinolophus rouxi.

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

Acoustic Stimulation↗

Activation of stress signaling molecules in bat brain during arousal from hibernation.

Induction of glucose-regulated proteins (GRPs) is a ubiquitous intracellular response to stresses such as hypoxia, glucose starvation and acidosis. The induction of GRPs offers some protection against these stresses in vitro, but the specific role of GRPs in vivo remains unclear. Hibernating bats present a good in vivo model to address this question. The bats must overcome local high oxygen demand in tissue by severe metabolic stress during arousal thermogenesis. We used brain tissue of a temperate bat Rhinolopus ferrumequinum to investigate GRP induction by high metabolic oxygen demand and to identify associated signaling molecules. We found that during 30 min of arousal, oxygen consumption increased from nearly zero to 11.9/kg/h, which was about 8.7-fold higher than its active resting metabolic rate. During this time, body temperature rose from 7 degrees C to 35 degrees C, and levels of TNF-alpha and lactate in brain tissue increased 2-2.5-fold, indicating a high risk of oxygen shortage. Concomitantly, levels of GRP75, GRP78 and GRP94 increased 1.5-1.7-fold. At the same time, c-Jun N-terminal protein kinase (JNK) activity increased 6.4-fold, and extracellular signal-regulated protein kinase (ERK) activity decreased to a similar degree (6.1-fold). p38 MAPK activity was very low and remained unchanged during arousal. In addition, survival signaling molecules protein kinase B (Akt) and protein kinase C (PKC) were activated 3- and 5-fold, respectively, during arousal. Taken together, our results showed that bat brain undergoes high oxygen demand during arousal from hibernation. Up-regulation of GRP proteins and activation of JNK, PKCgamma and Akt may be critical for neuroprotection and the survival of bats during the repeated process.

Animals↗

Life history, ecology and longevity in bats.

The evolutionary theory of aging predicts that life span should decrease in response to the amount of mortality caused by extrinsic sources. Using this prediction, we selected six life history and ecological factors to use in a comparative analysis of longevity among 64 bat species. On average, the maximum recorded life span of a bat is 3.5 times greater than a non-flying placental mammal of similar size. Records of individuals surviving more than 30 years in the wild now exist for five species. Univariate and multivariate analyses of species data, as well as of phylogenetically independent contrasts obtained using a supertree of Chiroptera, reveal that bat life span significantly increases with hibernation, body mass and occasional cave use, but decreases with reproductive rate and is not influenced by diet, colony size or the source of the record. These results are largely consistent with extrinsic mortality risk acting as a determinant of bat longevity. Nevertheless, the strong association between life span and both reproductive rate and hibernation also suggests that bat longevity is strongly influenced by seasonal allocation of non-renewable resources to reproduction. We speculate that hibernation may provide a natural example of caloric restriction, which is known to increase longevity in other mammals.

Animals↗