A chromosome-level genome assembly of an avivorous bat species (Nyctalus aviator).

Currently, three carnivorous bat species, namely Ia io, Nyctalus lasiopterus, and Nyctalus aviator, are known to actively prey on seasonal migratory birds (hereinafter referred to as "avivorous bats"). However, the absence of reference genomes impedes a thorough comprehension of the molecular adaptations of avivorous bat species. Herein, we present the high-quality chromosome-scale reference genome of N. aviator based on PacBio subreads, DNBSEQ short-reads and Hi-C sequencing data. The genome assembly size of N. aviator is 1.77 Gb, with a scaffold N50 of 102 Mb, of which 99.8% assembly was anchored into 21 pseudo-chromosomes. After masking 635.1 Mb repetitive sequences, a total of 19,412 protein-coding genes were identified, of which 99.3% were functionally annotated. The genome assembly and gene prediction reached 96.1% and 96.1% completeness of Benchmarking Universal Single-Copy Orthologs (BUSCO), respectively. This chromosome-level reference genome of N. aviator fills a gap in the existing information on the genomes of carnivorous bats, especially avivorous ones, and will be valuable for mechanism of adaptations to dietary niche expansion in bat species.

Contrasting laboratory and field outcomes of bat-moth interactions.

Predator-prey interactions are important but difficult to study in the field. Therefore, laboratory studies are often used to examine the outcomes of predator-prey interactions. Previous laboratory studies have shown that moth hearing and ultrasound production can help prey avoid being eaten by bats. We report here that laboratory behavioural outcomes may not accurately reflect the outcomes of field bat-moth interactions. We tested the success rates of two bat species capturing moths with distinct anti-bat tactics using behavioural experiments. We compared the results with the dietary composition of field bats using next-generation DNA sequencing. Rhinolophus episcopus and Rhinolophus osgoodi had a lower rate of capture success when hunting for moths that produce anti-bat clicks than for silent eared moths and earless moths. Unexpectedly, the success rates of the bats capturing silent eared moths and earless moths did not differ significantly from each other. However, the field bats had a higher proportion of silent eared moths than that of earless moths and that of clicking moths in their diets. The difference between the proportions of silent eared moths and earless moths in the bat diets can be explained by the difference between their abundance in bat foraging habitats. These findings suggest that moth defensive tactics, bat countertactics and moth availability collectively shape the diets of insectivorous bats. This study illustrates the importance of using a combination of behavioural experiments and molecular genetic techniques to reveal the complex interactions between predators and prey in nature.

Protocol for simulating foraging behavior strategy and dual-choice predatory sensory cue experiments in aerial-hawking bats.

The nocturnal and cryptic nature of bats makes it difficult to determine their foraging behavior and predatory sensory cues. Here, we present a protocol for determining the behavioral strategies and sensory cues of bat predation on airborne prey. We describe steps for indoor foraging behavior simulation, dual-choice acoustic playback, and visual presentation experiments. This protocol was used to study the avivorous bat, Ia io, but it can be adapted for studying other aerial-hawking bats and even other taxa. For complete details on the use and execution of this protocol, please refer to Gong et al. (2022).1.

Untargeted metabolomics of the cochleae from two laryngeally echolocating bats.

High-frequency hearing is regarded as one of the most functionally important traits in laryngeally echolocating bats. Abundant candidate hearing-related genes have been identified to be the important genetic bases underlying high-frequency hearing for laryngeally echolocating bats, however, extensive metabolites presented in the cochleae have not been studied. In this study, we identified 4,717 annotated metabolites in the cochleae of two typical laryngeally echolocating bats using the liquid chromatography-mass spectroscopy technology, metabolites classified as amino acids, peptides, and fatty acid esters were identified as the most abundant in the cochleae of these two echolocating bat species, Rhinolophus sinicus and Vespertilio sinensis. Furthermore, 357 metabolites were identified as significant differentially accumulated (adjusted p-value <0.05) in the cochleae of these two bat species with distinct echolocating dominant frequencies. Downstream KEGG enrichment analyses indicated that multiple biological processes, including signaling pathways, nervous system, and metabolic process, were putatively different in the cochleae of R. sinicus and V. sinensis. For the first time, this study investigated the extensive metabolites and associated biological pathways in the cochleae of two laryngeal echolocating bats and expanded our knowledge of the metabolic molecular bases underlying high-frequency hearing in the cochleae of echolocating bats.

Full-Length Transcriptome of the Great Himalayan Leaf-Nosed Bats (Hipposideros armiger) Optimized Genome Annotation and Revealed the Expression of Novel Genes.

The Great Himalayan Leaf-nosed bat (Hipposideros armiger) is one of the most representative species of all echolocating bats and is an ideal model for studying the echolocation system of bats. An incomplete reference genome and limited availability of full-length cDNAs have hindered the identification of alternatively spliced transcripts, which slowed down related basic studies on bats' echolocation and evolution. In this study, we analyzed five organs from H. armiger for the first time using PacBio single-molecule real-time sequencing (SMRT). There were 120 GB of subreads generated, including 1,472,058 full-length non-chimeric (FLNC) sequences. A total of 34,611 alternative splicing (AS) events and 66,010 Alternative Polyadenylation (APA) sites were detected by transcriptome structural analysis. Moreover, a total of 110,611 isoforms were identified, consisting of 52% new isoforms of known genes and 5% of novel gene loci, as well as 2112 novel genes that have not been annotated before in the current reference genome of H. armiger. Furthermore, several key novel genes, including Pol, RAS, NFKB1, and CAMK4, were identified as being associated with nervous, signal transduction, and immune system processes, which may be involved in regulating the auditory nervous perception and immune system that helps bats to regulate in echolocation. In conclusion, the full-length transcriptome results optimized and replenished existing H. armiger genome annotation in multiple ways and offer advantages for newly discovered or previously unrecognized protein-coding genes and isoforms, which can be used as a reference resource.

Adaptive temporal patterns of echolocation and flight behaviors used to fly through varied-sized windows by 2 species of high duty cycle bats.

As actively sensing animals guided by acoustic information, echolocating bats must adapt their vocal-motor behavior to various environments and behavioral tasks. Here, we investigated how the temporal patterns of echolocation and flight behavior were adjusted in 2 species of bats with a high duty cycle (HDC) call structure, Rhinolophus ferrumequinum and Hipposideros armiger, when they flew along a straight corridor and then passed through windows of 3 different sizes. We also tested whether divergence existed in the adaptations of the 2 species. Both H. armiger and R. ferrumequinum increased their call rates by shortening the pulse duration and inter-pulse interval for more rapid spatial sampling of the environment when flying through smaller windows. Bats produced more sonar sound groups (SSGs) while maintaining a stable proportion of calls that made up SSGs during approaches to smaller windows. The 2 species showed divergent adjustment in flight behavior across 3 different window sizes. Hipposideros armiger reduced its flight speed to pass through smaller windows while R. ferrumequinum increased its flight speed. Our results suggest that these 2 species of HDC bats adopt similar acoustic timing patterns for different tasks although they performed different flight behaviors.

Seasonal variation of population and individual dietary niche in the avivorous bat, Ia io.

The variation in niche breadth can affect how species respond to environmental and resource changes. However, there is still no clear understanding of how seasonal variability in food resources impacts the variation of individual dietary diversity, thereby affecting the dynamics of a population's dietary niche breadth. Optimal foraging theory (OFT) and the niche variation hypothesis (NVH) predict that when food resources are limited, the population niche breadth will widen or narrow due to increased within-individual dietary diversity and individual specialization or reduced within-individual dietary diversity, respectively. Here, we used DNA metabarcoding to examine the composition and seasonality of diets of the avivorous bat Ia io. Furthermore, we investigated how the dietary niches changed among seasons and how the population niche breadth changed when the availability of insect resources was reduced in autumn. We found that there was differentiation in dietary niches among seasons and a low degree of overlap, and the decrease of insect resource availability and the emergence of ecological opportunities of nocturnal migratory birds might drive dietary niche shifts toward birds in I. io. However, the population's dietary niche breadth did not broaden by increasing the within-individual dietary diversity or individual specialization, but rather became narrower by reducing dietary diversity via predation on bird resources that served as an ecological opportunity when insect resources were scarce in autumn. Our findings were consistent with the predictions of OFT, because birds as prey for bats provided extremely different resources from those of insects in size and nutritional value. Our work highlights the importance of size and quality of prey resources along with other factors (i.e., physiological, behavioral, and life-history traits) in dietary niche variation.

Greater Horseshoe Bats Recognize the Sex and Individual Identity of Conspecifics from Their Echolocation Calls.

The echolocation calls of bats are mainly used for navigation and foraging; however, they may also contain social information about the emitter and facilitate social interactions. In this study, we recorded the echolocation calls of greater horseshoe bats (Rhinolophus ferrumequinum) and analyzed the acoustic parameter differences between the sexes and among individuals. Then, we performed habituation-discrimination playback experiments to test whether greater horseshoe bats could recognize the sex and individual identity of conspecifics from their echolocation calls. The results showed that there were significant differences in the echolocation call parameters between sexes and among individuals. When we switched playback files from a habituated stimuli to a dishabituated stimuli, the tested bats exhibited obvious behavioral responses, including nodding, ear or body movement, and echolocation emission. The results showed that R. ferrumequinum can recognize the sex and individual identity of conspecifics from their echolocation calls alone, which indicates that the echolocation calls of R. ferrumequinum may have potential communication functions. The results of this study improve our understanding of the communication function of the echolocation calls of bats.

Behavioral innovation and genomic novelty are associated with the exploitation of a challenging dietary opportunity by an avivorous bat.

Foraging on nocturnally migrating birds is one of the most challenging foraging tasks in the animal kingdom. Only three bat species (e.g., Ia io) known to date can prey on migratory birds. However, how these bats have exploited this challenging dietary niche remains unknown. Here, we demonstrate that I. io hunts at the altitude of migrating birds during the bird migration season. The foraging I. io exhibited high flight altitudes (up to 4945 m above sea level) and high flight speeds (up to 143.7 km h-1). I. io in flight can actively prey on birds in the night sky via echolocation cues. Genes associated with DNA damage repair, hypoxia adaptation, biting and mastication, and digestion and metabolism have evolved to adapt to this species' avivorous habits. Our results suggest that the evolution of behavioral innovation and genomic novelty are associated with the exploitation of challenging dietary opportunities.

The role of the gut microbiota in the dietary niche expansion of fishing bats.

BACKGROUND: Due to its central role in animal nutrition, the gut microbiota is likely a relevant factor shaping dietary niche shifts. We analysed both the impact and contribution of the gut microbiota to the dietary niche expansion of the only four bat species that have incorporated fish into their primarily arthropodophage diet. RESULTS: We first compared the taxonomic and functional features of the gut microbiota of the four piscivorous bats to that of 11 strictly arthropodophagous species using 16S rRNA targeted amplicon sequencing. Second, we increased the resolution of our analyses for one of the piscivorous bat species, namely Myotis capaccinii, and analysed multiple populations combining targeted approaches with shotgun sequencing. To better understand the origin of gut microorganisms, we also analysed the gut microbiota of their fish prey (Gambusia holbrooki). Our analyses showed that piscivorous bats carry a characteristic gut microbiota that differs from that of their strict arthropodophagous counterparts, in which the most relevant bacteria have been directly acquired from their fish prey. This characteristic microbiota exhibits enrichment of genes involved in vitamin biosynthesis, as well as complex carbohydrate and lipid metabolism, likely providing their hosts with an enhanced capacity to metabolise the glycosphingolipids and long-chain fatty acids that are particularly abundant in fish. CONCLUSIONS: Our results depict the gut microbiota as a relevant element in facilitating the dietary transition from arthropodophagy to piscivory.

Value of next-generation sequencing in early diagnosis of patients with tuberculous meningitis.

OBJECTIVES: To assess the value of next-generation sequencing (NGS) technology in early diagnosis of patients with tuberculous meningitis (TBM). METHODS: 56 patients with clinically suspected TBM who came to Shandong Provincial Chest Hospital from February 2, 2018 to August 2, 2018 were prospectively included, and the clinical diagnosis and treatment outcomes were followed up. NGS was performed for the cerebrospinal fluid specimens submitted for test on the BGISEQ-100 platform of Tianjin Huada Gene Research Institute and the obtained pathogen sequences were compared with the pathogen data to get the final results. The NGS results were positive for detecting the unique matching sequence of the Mycobacterium tuberculosis (MTB) complex and negative for no unique matching sequence. Patients confirmed with TBM should have at least one of the following four items: cerebrospinal fluid MTB culture positive, smear positive, Xpert MTB/RIF test positive, or MTB nucleic acid polymerase chain reaction (PCR) test positive; clinically diagnosed patients were those with clinically suspected TBM and effective anti-tuberculosis treatment; non-TBM patients were those with other pathogenic basis or clinical exclusion of TBM. The sensitivity and specificity of NGS in early diagnosis of TBM were analyzed. RESULTS: 22 patients were confirmed with TBM, of which 13 were positive for Xpert MTB/RIF test, 6 were positive for cerebrospinal fluid MTB culture, 5 were positive for MTB nucleic acid PCR test, 12 patients were clinically diagnosed with TBM, and there were 16 cases of non-TBM patients. Among confirmed and clinically diagnosed patients, 20 cases of MTB complex were detected by NGS technology, with a sensitivity of 58.8% (20/34) and specificity of 100% (16/16). Among confirmed patients, the sensitivity of NGS was 63.6% (14/22). Of the 50 specimens that were simultaneously subjected to traditional methods, Xpert MTB/RIF test and NGS, the specificity of the three methods was 100% (16/16) based on clinical diagnosis, and the sensitivity was 29.4% (10/34), 38.2% (13/34), and 58.8% (20/34) respectively. The difference of sensitivity between the first two detection methods and NGS was statistically significant (McNemar test, p = 0.013, x2 = 5.786 and p = 0.065, x2 = 3.273). The sensitivity of traditional methods combined with NGS was as high as 82.4% (28/34). CONCLUSIONS: NGS technology could rapidly detect the MTB complex in cerebrospinal fluid with significant sensitivity and specificity, which could be used as an early diagnosis index of TBM. NGS combined with MTB culture could increase the detection rate.

Territorial calls of the bat Hipposideros armiger may encode multiple types of information: body mass, dominance rank and individual identity.

In highly vocal species, territorial aggression is often accompanied using vocalizations. These vocalizations can play a critical role in determining the outcome of male-male agonistic interactions. For this, vocalizations of contestants must contain information that is indicative of each competitor's fighting ability as well as its identity, and also contestants must be able to perceive information about the physical attributes, quality and identity of the vocalizer. Here, we used adult male Great Himalayan leaf-nosed bats (Hipposideros armiger) to test whether territorial calls encoded honest information about a caller's physical attributes, quality and individual identity. We did this by exploring the relationships between territorial calls and two potential indices of fighting ability: body mass and dominance rank. Using synchronized audio-video recording, we monitored bat territorial calls and dominance rank of 16 adult male H. armiger in the laboratory. Additionally, habituation-dishabituation playback experiments were performed to test for vocal discrimination. Results showed that body mass was negatively related to minimum frequency and positively related to syllable duration. Dominance score was also negatively related to minimum frequency and positively related to peak frequency. Furthermore, a discriminant function analysis suggested that territorial calls encode an individual signature. Therefore, our data show that males have the ability to utilize this vocal individual signature to discriminate between vocalizing males. In short, territorial calls of male H. armiger contain information about body mass, dominance rank and individual identity, and contestants are probably capable of perceiving this information and may use it to make appropriate decisions during agonistic interactions.

Correlation of skull morphology and bite force in a bird-eating bat (Ia io; Vespertilionidae).

BACKGROUND: Genetic and ecological factors influence morphology, and morphology is compatible with function. The morphology and bite performance of skulls of bats show a number of characteristic feeding adaptations. The great evening bat, Ia io (Thomas, 1902), eats both insects and birds (Thabah et al. J Mammal 88: 728-735, 2007), and as such, it is considered to represent a case of dietary niche expansion from insects to birds. How the skull morphology or bite force in I. io are related to the expanded diet (that is, birds) remains unknown. We used three-dimensional (3D) geometry of the skulls and measurements of bite force and diets from I. io and 13 other species of sympatric or closely related bat species to investigate the characteristics and the correlation of skull morphology and bite force to diets. RESULTS: Significant differences in skull morphology and bite force among species and diets were observed in this study. Similar to the carnivorous bats, bird-eaters (I. io) differed significantly from insectivorous bats; I. io had a larger skull size, taller crania, wider zygomatic arches, shorter but robust mandibles, and larger bite force than the insectivores. The skull morphology of bats was significantly associated with bite force whether controlling for phylogeny or not, but no significant correlations were found between diets and the skulls, or between diets and residual bite force, after controlling for phylogeny. CONCLUSIONS: These results indicated that skull morphology was independent of diet, and phylogeny had a greater impact on skull morphology than diet in these species. The changes in skull size and morphology have led to variation in bite force, and finally different bat species feeding on different foods. In conclusion, I. io has a larger skull size, robust mandibles, shortened dentitions, longer coronoid processes, expanded angular processes, low condyles, and taller cranial sagittal crests, and wider zygomatic arches that provide this species with mechanical advantages; their greater bite force may help them use larger and hard-bodied birds as a dietary component.

The roles of morphological traits, resource variation and resource partitioning associated with the dietary niche expansion in the fish-eating bat Myotis pilosus.

Niche expansion and shifts are involved in the response and adaptation to environmental changes. However, it is unclear how niche breadth evolves and changes toward higher-quality resources. Myotis pilosus is both an insectivore and a piscivore. We examined the dietary composition and seasonality in M. pilosus and the closely related Myotis fimbriatus using next-generation DNA sequencing. We tested whether resource variation or resource partitioning help explain the dietary expansion from insects to fish in M. pilosus. While diet composition and diversity varied significantly between summer and autumn, the proportion of fish-eating individuals did not significantly change between seasons in M. pilosus. Dietary overlap between M. pilosus and M. fimbriatus during the same seasons was much higher than within individual species across seasons. We recorded a larger body size, hind foot length, and body mass in M. pilosus than in M. fimbriatus and other insectivorous trawling bats from China. Similar morphological differences were found between worldwide fishing bats and nonfishing trawling bats. Our results suggest that variation in insect availability or interspecific competition may not play important roles in the dietary expansion from insects to fish in M. pilosus. Myotis pilosus has morphological advantages that may help it use fish as a diet component. The morphological advantage promoting dietary niche evolution toward higher quality resources may be more important than variation in the original resource and the effects of interspecific competition.

Bats adjust temporal parameters of echolocation pulses but not those of communication calls in response to traffic noise.

Many studies based on acute short-term noise exposure have demonstrated that animals can adjust their vocalizations in response to ambient noise. However, the effects of chronic noise over a relatively long time scale of multiple days remain largely unclear. Bats rely mainly on acoustic signals for perception of environmental and social communication. Nearly all previous studies on noise-induced vocal adjustments have focused on echolocation pulse sounds. Relatively little is known regarding the effects of noise on social communication calls. Here, we examined the dynamic changes in the temporal parameters of echolocation and communication vocalizations of Vespertilio sinensis when exposed to traffic noise over multiple days. We found that the bats started to modify their echolocation vocalizations on the fourth day of noise exposure, with an increase of 42-91% in the total number of pulse sequences per day. Under noisy conditions, the number of pulses within a pulse sequence decreased by an average of 17.2%, resulting in a significantly slower number of pulses/sequence (P < 0.001). However, there was little change in the duration of a pulse sequence. These parameters were not significantly adjusted in most communication vocalizations under the noise condition (all P > 0.05), except that the duration decreased and the number of syllables/sequences increased in 1 type of communicative vocalization (P < 0.05). This study suggests that bats routinely adjust temporal parameters of echolocation but rarely of communication vocalizations in response to noise condition.

Bats increase vocal amplitude and decrease vocal complexity to mitigate noise interference during social communication.

Natural background noises are common in the acoustic environments in which most organisms have evolved. Therefore, the vocalization and sound perception systems of vocal animals are inherently equipped to overcome natural background noise. Human-generated noises, however, pose new challenges that can hamper audiovocal communication. The mechanisms animals use to cope with anthropogenic noise disturbances have been extensively explored in a variety of taxa. Bats emit echolocation pulses primarily to orient, locate and navigate, while social calls are used to communicate with conspecifics. Previous studies have shown that bats alter echolocation pulse parameters in response to background noise interference. In contrast to high-frequency echolocation pulses, relatively low-frequency components within bat social calls overlap broadly with ambient noise frequencies. However, how bats structure their social calls in the presence of anthropogenic noise is not known. Here, we hypothesized that bats leverage vocal plasticity to facilitate vocal exchanges within a noisy environment. To test this hypothesis, we subjected the Asian particolored bat, Vespertilio sinensis, to prerecorded traffic noise. We observed a significant decrease in vocal complexity (i.e., an increased frequency of monosyllabic calls) in response to traffic noise. However, an increase in the duration and frequency of social calls, as have been observed in other species, was not evident. This suggests that signal simplification may increase communication efficacy in noisy environments. Moreover, V. sinensis also increased call amplitude in response to increased traffic noise, consistent with the predictions of the Lombard effect.

Great Himalayan leaf-nosed bats modify vocalizations to communicate threat escalation during agonistic interactions.

Bats vocalize extensively within various social contexts. Nevertheless, studies of agonistic interactions, associating vocalizations signalling the emotional state of a caller with individual signatures during aggressive vocalizations remain scarce. Here, we examined whether male Great Himalayan leaf-nosed bats (Hipposideros armiger) modify their aggressive vocalizations during agonistic interactions depending on the level of aggression, and whether these vocalizations encode individual signatures. We applied a cost-benefit analysis to audio-video recordings of 50 dyadic agonistic interactions to categorize displays into two levels of aggression intensity (low aggression: bared teeth, slightly pulled up body and/or wings; high aggression: rapidly flapping wings, punching, biting). Male H. armiger used graded visual agonistic displays accompanied by bent upward frequency modulation (bUFM) vocalizations to defend their roosting territories. At high aggression intensities, males decreased the minimum frequency of aggressive calls and increased the frequency bandwidth. Males also transferred energy from the second harmonic to the first harmonic as the threat escalated. These systematic modifications of acoustic parameters as aggressive intensity fluctuated corresponded to prosodic modifications in human speech, indicating that emotion-related acoustic cues are a common feature of acoustic communication in mammals. In addition, we found that the aggressive calls of eight adult males encoded discriminable signatures, and that males could discriminate among individuals based on these aggressive calls. Such discrimination is probably useful for individual identification among rival neighbours.

Geographical variation in the echolocation calls of bent-winged bats, Miniopterus fuliginosus.

Evolutionary biologists had a long-standing interest in the evolutionary forces underlying geographical variation in the acoustic signals of animals. However, the evolutionary forces driving acoustic variation are still unclear. In this study, we quantified the geographical variation in the peak frequencies of echolocation calls in eight Miniopterus fuliginosus bat colonies, and assessed the forces that drive acoustic divergence. Our results demonstrated that seven of the colonies had very similar peak frequencies, while only one colony was significantly higher than the others. This similarity in echolocation call frequency among the seven colonies was likely due to frequent dispersal and migration, leading to male-mediated infiltration of nuclear genes. This infiltration enhances gene flow and weakens ecological selection, and also increases interactions in the presence of conspecifics. Significant correlations were not observed between acoustic distances and morphological distances, climatic differences, geographic distances or mtDNA genetic distances. However, variation in acoustic distances was significantly positive correlated with nDNA genetic distance, even after controlling for geographic distance. Interestingly, the relationship between call divergence and genetic distance was no longer significant after excluding the colony with the highest call frequency, which may be due to the minimal genetic distance among the other seven colonies. The highest frequencies of echolocation calls observed in the one colony may be shaped by selection pressure due to loud background noise in the area. Taken together, these results suggest that geographic divergence of echolocation calls may not be subject to genetic drift, but rather, that the strong selective pressure induced by background noise may lead to acoustic and genetic differentiation between JXT and the other colonies.

Stereotypy and variability of social calls among clustering female big-footed myotis (Myotis macrodactylus).

Echolocating bats have developed advanced auditory perception systems, predominantly using acoustic signaling to communicate with each other. They can emit a diverse range of social calls in complex behavioral contexts. This study examined the vocal repertoire of five pregnant big-footed myotis bats (Myotis macrodactylus). In the process of clustering, the last individual to return to the colony (LI) emitted social calls that correlated with behavior, as recorded on a PC-based digital recorder. These last individuals could emit 10 simple monosyllabic and 27 complex multisyllabic types of calls, constituting four types of syllables. The social calls were composed of highly stereotyped syllables, hierarchically organized by a common set of syllables. However, intra-specific variation was also found in the number of syllables, syllable order and patterns of syllable repetition across call renditions. Data were obtained to characterize the significant individual differences that existed in the maximum frequency and duration of calls. Time taken to return to the roost was negatively associated with the diversity of social calls. Our findings indicate that variability in social calls may be an effective strategy taken by individuals during reintegration into clusters of female M. macrodactylus.

The first record of Ia io Thomas, 1902 (Mammalia: Chiroptera: Vespertilionidae) from the Sundaic Subregion, with a description of a new subspecies from peninsular Thailand.

The Great evening bat Ia io Thomas, 1902, previously considered as an endemic to the Indochinese subregion, is reported from the Sundaic subregion for the first time based on specimens collected from three localities in Surat Thani Province and Phang Nga Province, peninsular Thailand. It is described herein as a new subspecies based on its substantially larger body and skull size. The mitochondrial COI and cytochrome b genes reveal that the new subspecies has a genetic distance of 1.89% and 1.65%, respectively, from the nominate subspecies. Echolocation calls comprise four harmonics, with the maximum energy in the first harmonic (fmaxe) of 23.6-27.4 kHz. Notes on the population size as well as roosting and foraging behaviour are also provided.