Molecular signatures of longevity: Insights from cross-species comparative studies.

Much of the current research on longevity focuses on the aging process within a single species. Several molecular players (e.g. IGF1 and MTOR), pharmacological compounds (e.g. rapamycin and metformin), and dietary approaches (e.g. calorie restriction and methionine restriction) have been shown to be important in regulating and modestly extending lifespan in model organisms. On the other hand, natural lifespan varies much more significantly across species. Within mammals alone, maximum lifespan differs more than 100 fold, but the underlying regulatory mechanisms remain poorly understood. Recent comparative studies are beginning to shed light on the molecular signatures associated with exceptional longevity. These include genome sequencing of microbats, naked mole rat, blind mole rat, bowhead whale and African turquoise killifish, and comparative analyses of gene expression, metabolites, lipids and ions across multiple mammalian species. Together, they point towards several putative strategies for lifespan regulation and cancer resistance, as well as the pathways and metabolites associated with longevity variation. In particular, longevity may be achieved by both lineage-specific adaptations and common mechanisms that apply across the species. Comparing the resulting cross-species molecular signatures with the within-species lifespan extension strategies will improve our understanding of mechanisms of longevity control and provide a starting point for novel and effective interventions.

Slow aging in mammals-Lessons from African mole-rats and bats.

Traditionally, the main mammalian models used in aging research have been mice and rats, i.e. short-lived species that obviously lack effective maintenance mechanisms to keep their soma in a functional state for prolonged periods of time. It is doubtful that life-extending mechanisms identified only in such short-lived species adequately reflect the diversity of longevity pathways that have naturally evolved in mammals, or that they have much relevance for long-lived species such as humans. Therefore, some complementary, long-lived mammalian models have been introduced to aging research in the past 15-20 years, particularly naked mole-rats (and to a lesser extent also other mole-rats) and bats. Here, I summarize and compare the most important results regarding various aspects of aging - oxidative stress, molecular homeostasis and repair, and endocrinology - that have been obtained from studies using these new mammalian models of high longevity. I argue that the inclusion of these models was an important step forward, because it drew researchers' attention to certain oversimplifications of existing aging theories and to several features that appear to be universal components of enhanced longevity in mammals. However, even among mammals with high longevity, considerable variation exists with respect to other candidate mechanisms that also must be taken into account if inadequate generalizations are to be avoided.

Prenatal allometric trajectories and the developmental basis of postcranial phenotypic diversity in bats (Chiroptera).

Most morphological and physiological adaptations associated with bat flight are concentrated in the postcranium, reflecting strong functional demands for flight performance. Despite an association between locomotory diversity and trophic differentiation, postcranial morphological diversity in bats remains largely unexplored. Evolutionary developmental biology is a novel approach providing a link between the analysis of genotypic and phenotypic variation resulting from selective pressures. To quantify the morphological diversity of the postcranium in bats and to explore its developmental basis, we reconstructed the postcranial allometric trajectories of nine bat species from different prenatal developmental series, representing five families and both suborders. We tested for allometric growth in Chiroptera and also quantified levels of allometric disparity and inter-trajectory distances. Using a phylogenetic scaffold, we assessed whether ontogenetic differences reflect evolutionary relationships. We found significant allometric growth trajectories in almost all species. Interspecific trajectory distances showed lower variance within Yinpterochiroptera than within Yangochiroptera and between suborders. Each suborder occupied nonoverlapping sections of allometric space, showing changes in the growth rates of specific bones for each suborder. The allometry-corrected disparity was significantly higher in larger species. Statistically significant phylogenetic signal in our results suggests that there is an ontogenetic basis for the postcranial morphological diversity in modern bats. Ancestral state reconstruction also showed an increase in the amount of change in shape with size in the larger species studied. We hypothesize that differences in allometric patterns among bat taxa may reflect a size-dependent evolutionary constraint, whereby variability in body size and allometric patterns are associated.

Bat Accelerated Regions Identify a Bat Forelimb Specific Enhancer in the HoxD Locus.

The molecular events leading to the development of the bat wing remain largely unknown, and are thought to be caused, in part, by changes in gene expression during limb development. These expression changes could be instigated by variations in gene regulatory enhancers. Here, we used a comparative genomics approach to identify regions that evolved rapidly in the bat ancestor, but are highly conserved in other vertebrates. We discovered 166 bat accelerated regions (BARs) that overlap H3K27ac and p300 ChIP-seq peaks in developing mouse limbs. Using a mouse enhancer assay, we show that five Myotis lucifugus BARs drive gene expression in the developing mouse limb, with the majority showing differential enhancer activity compared to the mouse orthologous BAR sequences. These include BAR116, which is located telomeric to the HoxD cluster and had robust forelimb expression for the M. lucifugus sequence and no activity for the mouse sequence at embryonic day 12.5. Developing limb expression analysis of Hoxd10-Hoxd13 in Miniopterus natalensis bats showed a high-forelimb weak-hindlimb expression for Hoxd10-Hoxd11, similar to the expression trend observed for M. lucifugus BAR116 in mice, suggesting that it could be involved in the regulation of the bat HoxD complex. Combined, our results highlight novel regulatory regions that could be instrumental for the morphological differences leading to the development of the bat wing.

Metal and metalloid exposure and oxidative status in free-living individuals of Myotis daubentonii.

Metal elements, ubiquitous in the environment, can cause negative effects in long-lived organisms even after low but prolonged exposure. Insectivorous bats living near metal emission sources can be vulnerable to such contaminants. Although it is known that bats can bioaccumulate metals, little information exists on the effects of metal elements on their physiological status. For example, oxidative status markers are known to vary after detoxification processes and immune reactions. Here, for two consecutive summers, we sampled individuals from a natural population of the insectivorous bat, Myotis daubentonii, inhabiting a site close to a metal emission source. We quantified metals and metalloids (As, Ca, Cd, Co, Cu, Mn, Ni, Pb, Se, Zn) from individual fecal pellets. We measured enzymatic antioxidants (GP, CAT, SOD), total glutathione (tGSH) and ratio between reduced and oxidized glutathione (GSH:GSSG) from their red blood cells together with biometrics, hematocrit and parasite prevalence. In general, metal concentrations in feces of M. daubentonii reflected the exposure to ambient contamination. This was especially evident in the higher concentrations of Cd, Co, Cu and Ni close to a smelter compared to a site with less contaminant exposure. Annual differences were also observed for most elements quantified. Sex-specific differences were observed for calcium and zinc excretion. SOD and CAT enzymatic activities were associated with metal levels (principal components of six metal elements), suggesting early signs of chronic stress in bats. The study also shows promise for the use of non-invasive sampling to assess the metal exposure on an individual basis and metal contamination in the environment.

Willingness to Pay for Conservation of Transborder Migratory Species: A Case Study of the Mexican Free-Tailed Bat in the United States and Mexico.

We estimated U.S. and Mexican citizens' willingness to pay (WTP) for protecting habitat for a transborder migratory species, the Mexican free-tailed bat (Tadarida brasiliensis mexicana), using the contingent valuation method. Few contingent valuation surveys have evaluated whether households in one country would pay to protect habitat in another country. This study addresses that gap. In our study, Mexican respondents were asked about their WTP for conservation of Mexican free-tailed bat habitat in Mexico and in the United States. Similarly, U.S. respondents were asked about their WTP for conservation in the United States and in Mexico. U.S. households would pay $30 annually to protect habitat in the United States and $24 annually to protect habitat in Mexico. Mexican households would pay $8 annually to protect habitat in Mexico and $5 annually to protect habitat in the United States. In both countries, these WTP amounts rose significantly for increasing the size of the bat population rather than simply stabilizing the current bat population. The ratio of Mexican household WTP relative to U.S. household WTP is nearly identical to that of Mexican household income relative to U.S. household income. This suggests that the perceived economic benefits received from the bats is similar in Mexico and the United States, and that scaling WTP by relative income in international benefit transfer may be plausible.

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

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

Postnatal growth and age estimation in Scotophilus kuhlii.

Adequate postnatal growth is important for young bats to develop skilled sensory and locomotor abilities, which are highly associated with their survival once independent. This study investigated the postnatal growth and development of Scotophilus kuhlii in captivity. An empirical growth curve was established, and the postnatal growth rate was quantified to derive an age-predictive equation. By further controlling the fostering conditions of twins, the differences in the development patterns between pups that received maternal care or were hand-reared were analyzed to determine whether the latter developed in the same manner as their maternally reared counterparts. Our results indicate that both forearm length and body mass increased rapidly and linearly during the first 4 weeks, after which the growth rate gradually decreased to reach a stable level. The first flight occurred at an average age of 39 days with a mean forearm length and body mass of 92.07% and 70.52% of maternal size, respectively. The developmental pattern of hand-reared pups, although similar to that of their maternally reared twin siblings, displayed a slightly faster growth rate in the 4th and 5th weeks. The heavier body mass of hand-reared pups during the pre-fledging period may cause higher wing loading, potentially influencing the flight performance and survival of the bats once independent.

Mercury in little brown bat (Myotis lucifugus) maternity colonies and its correlation with freshwater acidity in Nova Scotia, Canada.

Insectivorous little brown bats are exposed to elevated concentrations of mercury (Hg) through their preference for aquatic-based prey. Here we investigated spatial patterns of total Hg (THg) in fur from 10 little brown bat maternity colonies across Nova Scotia, and assessed relationships with the acidity of nearby lakes and rivers. Total Hg concentrations were measured in fur samples from 149 adult female little brown bats. Values showed significant variation among colonies (mean range 3.76-27.38 µg/g, dry weight), and 48% of individuals had Hg concentrations in excess of the 10 µg/g threshold associated with neurochemical changes in Chiroptera conspecifics (n = 26) from Virginia. Average surface water acidity parameters (pH and acid neutralization capacity) within an 8 km radius of each maternity roost showed strong negative associations with average colony fur THg concentrations. This suggests that freshwater acidity in foraging grounds explains much of the variation in average fur THg concentrations in little brown bat colonies. These findings highlight the significant role that water quality may have on Hg bioaccumulation within terrestrial species that feed on aquatic prey.

Unique expression patterns of multiple key genes associated with the evolution of mammalian flight.

Bats are the only mammals capable of true flight. Critical adaptations for flight include a pair of dramatically elongated hands with broad wing membranes. To study the molecular mechanisms of bat wing evolution, we perform genomewide mRNA sequencing and in situ hybridization for embryonic bat limbs. We identify seven key genes that display unique expression patterns in embryonic bat wings and feet, compared with mouse fore- and hindlimbs. The expression of all 5'HoxD genes (Hoxd9-13) and Tbx3, six known crucial transcription factors for limb and digit development, is extremely high and prolonged in the elongating wing area. The expression of Fam5c, a tumour suppressor, in bat limbs is bat-specific and significantly high in all short digit regions (the thumb and foot digits). These results suggest multiple genetic changes occurred independently during the evolution of bat wings to elongate the hand digits, promote membrane growth and keep other digits short. Our findings also indicate that the evolution of limb morphology depends on the complex integration of multiple gene regulatory networks and biological processes that control digit formation and identity, chondrogenesis, and interdigital regression or retention.

Effects of resistin on ovarian folliculogenesis and steroidogenesis in the vespertilionid bat, Scotophilus heathi.

The bat Scotophilus heathi exhibit prolonged anovulatory condition known as delayed ovulation coinciding with the period of extensive fat accumulation. The present study was undertaken to find out whether extensive accumulation of fat in S. heathi is responsible for suppression of ovarian activity by increasing production of adipokine resistin in the bat. This was achieved by (a) investigating variation in serum resistin level in relation to the changes in the body fat mass and (b) evaluating the effect of resistin treatment on ovarian activity with reference to steroid synthesis. An attempt was also made to determine whether resistin mediate its effects on ovary through signal transducer and activator of transcription 3 (STAT3) signaling mechanism. The results showed significant seasonal variation in serum resistin level with the peak level coinciding with the period of maximum fat accumulation, high circulating androgen level and period of anovulation. The treatment with resistin to the bat caused increase in androstenedione due to stimulatory effects on 3ß-hydroxysteroid dehydrogenase, but decrease in estradiol level due to inhibitory effect on aromatase. Resistin treatment increased androgen receptor protein together with increased insulin receptor but not through conventional luteinizing hormone receptor and steroidogenic acute regulatory protein mediated pathways. This study further showed that resistin treatment increases androstenedione synthesis and up-regulates insulin receptor in the ovary through STAT3 mediated pathways. These findings suggest that obese women through increased resistin synthesis may causes development of non-ovulatory antral follicles through insulin receptor signaling cascade.

Tacaribe virus causes fatal infection of an ostensible reservoir host, the Jamaican fruit bat.

Tacaribe virus (TCRV) was first isolated from 11 Artibeus species bats captured in Trinidad in the 1950s during a rabies virus surveillance program. Despite significant effort, no evidence of infection of other mammals, mostly rodents, was found, suggesting that no other vertebrates harbored TCRV. For this reason, it was hypothesized that TCRV was naturally hosted by artibeus bats. This is in stark contrast to other arenaviruses with known hosts, all of which are rodents. To examine this hypothesis, we conducted experimental infections of Jamaican fruit bats (Artibeus jamaicensis) to determine whether they could be persistently infected without substantial pathology. We subcutaneously or intranasally infected bats with TCRV strain TRVL-11573, the only remaining strain of TCRV, and found that low-dose (10(4) 50% tissue culture infective dose [TCID(50)]) inoculations resulted in asymptomatic and apathogenic infection and virus clearance, while high-dose (10(6) TCID(50)) inoculations caused substantial morbidity and mortality as early as 10 days postinfection. Uninoculated cage mates failed to seroconvert, and viral RNA was not detected in their tissues, suggesting that transmission did not occur. Together, these data suggest that A. jamaicensis bats may not be a reservoir host for TCRV.

A new species of Lonchophylla (Chiroptera, Phyllostomidae) from the Atlantic Forest of southeastern Brazil, with comments on L. bokermanni.

We examined Brazilian species of the nectar-feeding bats genus Lonchophylla (Phyllostomidae, Lonchophyllinae) to clarify the identity of Lonchophylla bokermanni and to determine the distribution of this and other species of Lonchophylla in eastern Brazil. As a result, we have found sufficient differences between Cerrado populations (including the type locality of L. bokermanni) and populations inhabiting the Atlantic Forest of southeastern Brazil,which warrant the treatment of the Atlantic Forest populations as a separate and new species. We describe this new species here as Lonchophylla peracchii, sp. nov. The new species appears to be restricted to the Atlantic Forest, whereas L. bokermanni is found only in Cerrado habitats.

The evolution of color vision in nocturnal mammals.

Nonfunctional visual genes are usually associated with species that inhabit poor light environments (aquatic/subterranean/nocturnal), and these genes are believed to have lost function through relaxed selection acting on the visual system. Indeed, the visual system is so adaptive that the reconstruction of intact ancestral opsin genes has been used to reject nocturnality in ancestral primates. To test these assertions, we examined the functionality of the short and medium- to long-wavelength opsin genes in a group of mammals that are supremely adapted to a nocturnal niche: the bats. We sequenced the visual cone opsin genes in 33 species of bat with diverse sensory ecologies and reconstructed their evolutionary history spanning 65 million years. We found that, whereas the long-wave opsin gene was conserved in all species, the short-wave opsin gene has undergone dramatic divergence among lineages. The occurrence of gene defects in the short-wave opsin gene leading to loss of function was found to directly coincide with the origin of high-duty-cycle echolocation and changes in roosting ecology in some lineages. Our findings indicate that both opsin genes have been under purifying selection in the majority bats despite a long history of nocturnality. However, when spectacular losses do occur, these result from an evolutionary sensory modality tradeoff, most likely driven by subtle shifts in ecological specialization rather than a nocturnal lifestyle. Our results suggest that UV color vision plays a considerably more important role in nocturnal mammalian sensory ecology than previously appreciated and highlight the caveat of inferring light environments from visual opsins and vice versa.

Elevated mercury exposure and neurochemical alterations in little brown bats (Myotis lucifugus) from a site with historical mercury contamination.

Despite evidence of persistent methylmercury (MeHg) contamination in the South River (Virginia, USA) ecosystem, there is little information concerning MeHg-associated neurological impacts in resident wildlife. Here we determined mercury (Hg) concentrations in tissues of insectivorous little brown bats (Myotis lucifugus) collected from a reference site and a MeHg-contaminated site in the South River ecosystem. We also explored whether neurochemical biomarkers (monoamine oxidase, MAO; acetylcholinesterase, ChE; muscarinic acetylcholine receptor, mAChR; N-methyl-D-aspartate receptor, NMDAR) previously shown to be altered by MeHg in other wildlife were associated with brain Hg levels in these bats. Concentrations of Hg (total and MeHg) in tissues were significantly higher (10-40 fold difference) in South River bats when compared to reference sites. Mean tissue mercury levels (71.9 ppm dw in liver, 7.14 ppm dw in brain, 132 ppm fw in fur) in the South River bats exceed (sub)-clinical thresholds in mammals. When compared to the South River bats, animals from the reference site showed a greater ability to demethylate MeHg in brain (33.1% of total Hg was MeHg vs. 65.5%) and liver (8.9% of total Hg was MeHg vs. 50.8%) thus suggesting differences in their ability to detoxify and eliminate Hg. In terms of Hg-associated neurochemical biomarker responses, interesting biphasic responses were observed with an inflection point between 1 and 5 ppm dw in the brain. In the reference bats Hg-associated decreases in MAO (r = -0.61; p < 0.05) and ChE (r = -0.79; p < 0.01) were found in a manner expected but these were not found in the bats from the contaminated site. Owing to high Hg exposures, differences in Hg metabolism, and the importance of the aforementioned neurochemicals in multiple facets of animal health, altered or perhaps even a lack of expected neurochemical responses in Hg-contaminated bats raise questions about the ecological and physiological impacts of Hg on the bat population as well as the broader ecosystem in the South River.

Population growth of Mexican free-tailed bats (Tadarida brasiliensis mexicana) predates human agricultural activity.

BACKGROUND: Human activities, such as agriculture, hunting, and habitat modification, exert a significant effect on native species. Although many species have suffered population declines, increased population fragmentation, or even extinction in connection with these human impacts, others seem to have benefitted from human modification of their habitat. Here we examine whether population growth in an insectivorous bat (Tadarida brasiliensis mexicana) can be attributed to the widespread expansion of agriculture in North America following European settlement. Colonies of T. b. mexicana are extremely large (~10(6) individuals) and, in the modern era, major agricultural insect pests form an important component of their food resource. It is thus hypothesized that the growth of these insectivorous bat populations was coupled to the expansion of agricultural land use in North America over the last few centuries. RESULTS: We sequenced one haploid and one autosomal locus to determine the rate and time of onset of population growth in T. b. mexicana. Using an approximate Maximum Likelihood method, we have determined that T. b. mexicana populations began to grow ~220 kya from a relatively small ancestral effective population size before reaching the large effective population size observed today. CONCLUSIONS: Our analyses reject the hypothesis that T. b. mexicana populations grew in connection with the expansion of human agriculture in North America, and instead suggest that this growth commenced long before the arrival of humans. As T. brasiliensis is a subtropical species, we hypothesize that the observed signals of population growth may instead reflect range expansions of ancestral bat populations from southern glacial refugia during the tail end of the Pleistocene.

Development of echolocation and communication vocalizations in the big brown bat, Eptesicus fuscus.

Big brown bats form large maternity colonies of up to 200 mothers and their pups. If pups are separated from their mothers, they can locate each other using vocalizations. The goal of this study was to systematically characterize the development of echolocation and communication calls from birth through adulthood to determine whether they develop from a common precursor at the same or different rates, or whether both types are present initially. Three females and their six pups were isolated from our captive breeding colony. We recorded vocal activity from postnatal day 1 to 35, both when the pups were isolated and when they were reunited with their mothers. At birth, pups exclusively emitted isolation calls, with a fundamental frequency range <20 kHz, and duration >30 ms. By the middle of week 1, different types of vocalizations began to emerge. Starting in week 2, pups in the presence of their mothers emitted sounds that resembled adult communication vocalizations, with a lower frequency range and longer durations than isolation calls or echolocation signals. During weeks 2 and 3, these vocalizations were extremely heterogeneous, suggesting that the pups went through a babbling stage before establishing a repertoire of stereotyped adult vocalizations around week 4. By week 4, vocalizations emitted when pups were alone were identical to adult echolocation signals. Echolocation and communication signals both appear to develop from the isolation call, diverging during week 2 and continuing to develop at different rates for several weeks until the adult vocal repertoire is established.

Developmental changes in ultrasonic vocalizations by infant Japanese echolocating bats, Pipistrellus abramus.

Developmental changes in vocalizations by Pipistrellus abramus were investigated during the first post-natal month. Vocalizations by pups on the day of birth were frequency-modulated ultrasounds from 30.0 ± 4.0 kHz to 19.3 ± 1.9 kHz with multiple harmonics. The terminal frequency of the second harmonic (TF(2)) of pup vocalizations corresponded to that of the fundamental (TF(1)) in adult bats (41.4 ± 2.6 kHz), suggesting that pup vocalizations can be easily detected by the mother. In addition, there are two types of infant vocalization: short duration echolocation precursor and long duration isolation calls, which showed separate developmental patterns over time.

Persistence of Multiple Paramyxoviruses in a Closed Captive Colony of Fruit Bats (Eidolon helvum).

Bats have been identified as the natural hosts of several emerging zoonotic viruses, including paramyxoviruses, such as Hendra and Nipah viruses, that can cause fatal disease in humans. Recently, African fruit bats with populations that roost in or near urban areas have been shown to harbour a great diversity of paramyxoviruses, posing potential spillover risks to public health. Understanding the circulation of these viruses in their reservoir populations is essential to predict and prevent future emerging diseases. Here, we identify a high incidence of multiple paramyxoviruses in urine samples collected from a closed captive colony of circa 115 straw-coloured fruit bats (Eidolon helvum). The sequences detected have high nucleotide identities with those derived from free ranging African fruit bats and form phylogenetic clusters with the Henipavirus genus, Pararubulavirus genus and other unclassified paramyxoviruses. As this colony had been closed for 5 years prior to this study, these results indicate that within-host paramyxoviral persistence underlies the role of bats as reservoirs of these viruses.

Longitudinal monitoring in Cambodia suggests higher circulation of alpha and betacoronaviruses in juvenile and immature bats of three species.

Recent studies suggest that coronaviruses circulate widely in Southeast Asian bat species and that the progenitors of the SARS-Cov-2 virus could have originated in rhinolophid bats in the region. Our objective was to assess the diversity and circulation patterns of coronavirus in several bat species in Southeast Asia. We undertook monthly live-capture sessions and sampling in Cambodia over 17 months to cover all phases of the annual reproduction cycle of bats and test specifically the association between their age and CoV infection status. We additionally examined current information on the reproductive phenology of Rhinolophus and other bat species presently known to occur in mainland southeast China, Vietnam, Laos and Cambodia. Results from our longitudinal monitoring (573 bats belonging to 8 species) showed an overall proportion of positive PCR tests for CoV of 4.2% (24/573) in cave-dwelling bats from Kampot and 4.75% (22/463) in flying-foxes from Kandal. Phylogenetic analysis showed that the PCR amplicon sequences of CoVs (n = 46) obtained clustered in Alphacoronavirus and Betacoronavirus. Interestingly, Hipposideros larvatus sensu lato harbored viruses from both genera. Our results suggest an association between positive detections of coronaviruses and juvenile and immature bats in Cambodia (OR = 3.24 [1.46-7.76], p = 0.005). Since the limited data presently available from literature review indicates that reproduction is largely synchronized among rhinolophid and hipposiderid bats in our study region, particularly in its more seasonal portions (above 16° N), this may lead to seasonal patterns in CoV circulation. Overall, our study suggests that surveillance of CoV in insectivorous bat species in Southeast Asia, including SARS-CoV-related coronaviruses in rhinolophid bats, could be targeted from June to October for species exhibiting high proportions of juveniles and immatures during these months. It also highlights the need to develop long-term longitudinal surveys of bats and improve our understanding of their ecology in the region, for both biodiversity conservation and public health reasons.