Search

Behavioral divergence among populations is common across taxonomic groups, still we know very little about anti-predator behaviors. Animal exposure to predation risk is variable in different ecological contexts. In addition, reproduction value of animals in different geographical regions usually varies. In this study, we tested whether cinereous tits Parus cinereus in different populations exhibited nest defense behaviors similar to those of nest or adult predators and whether their nest defense behaviors showed geographical variation. By using field dummy experiments, we observed tits’ nest defense behavior in nest predator common chipmunk Tamias sibiricus and red squirrel Sciurus vulgaris, adult predator Eurasian sparrowhawk Accipiter nisus and nonthreatening species Oriental turtle dove Streptopelia orientalis in the ZJ (44° N), DZ (31° N), and DLS (18° N) populations, respectively. The response scores varied significantly across the 4 dummies in ZJ-tits and DLS-tits but did not in DZ-tits. When facing the chipmunk, ZJ-tits showed the highest response score and DZ-tits showed the lowest response score. When facing the squirrel, ZJ-tits showed a higher response score than tits in the other 2 populations. However, tits among the 3 populations responded similarly to a sparrowhawk or dove with slight response behaviors. In addition, response scores to nest predators were positively correlated with brood size across the 3 populations, but no trend was found for sparrowhawks or doves. Our results indicated that the nest defense behaviors of cinereous tits have evolved in response to large-scale geographical variation in ecological contexts and reproduction value.

White-nose syndrome, a disease that is caused by the psychrophilic fungus Pseudogymnoascus destructans, has threatened several North America bat species with extinction. Recent studies have shown that East Asian bats are infected with P. destructans but show greatly reduced infections. While several factors have been found to contribute to these reduced infections, the role of specific microbes in limiting P. destructans growth remains unexplored. We isolated three bacterial strains with the ability to inhibit P. destructans, namely, Pseudomonas yamanorum GZD14026, Pseudomonas brenneri XRD11711 and Pseudomonas fragi GZD14479, from bats in China. Pseudomonas yamanorum, with the highest inhibition score, was selected to extract antifungal active substance. Combining mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy analyses, we identified the active compound inhibiting P. destructans as phenazine-1-carboxylic acid (PCA), and the minimal inhibitory concentration (MIC) was 50.12 mu g ml(-1). Whole genome sequencing also revealed the existence of PCA biosynthesis gene clusters. Gas chromatography-mass spectrometry (GC-MS) analysis identified volatile organic compounds. The results indicated that 10 ppm octanoic acid, 100 ppm 3-tert-butyl-4-hydroxyanisole (isoprenol) and 100 ppm 3-methyl-3-buten-1-ol (BHA) inhibited the growth of P. destructans. These results support that bacteria may play a role in limiting the growth of P. destructans on bats. National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31961123001, 31770403, 31570390, 31670390]; Jilin Provincial Natural Science Foundation [20180101272JC]; Program for Introducing Talents to Universities [B16011]; US National Science FoundationNational Science Foundation (NSF) [DEB-1911853] This work was supported by the National Natural Science Foundation of China (grant numbers 31961123001, 31770403, 31570390 and 31670390), Jilin Provincial Natural Science Foundation (grant number 20180101272JC), and the Program for Introducing Talents to Universities (B16011) and the US National Science Foundation (DEB-1911853).

Many territorial animals are less aggressive towards neighbours than to strangers, a phenomenon known as the ‘dear enemy effect’. However, there has been little research on changes in this behaviour in songbirds across the breeding season. In this study, we followed changes in the dear enemy effect during the breeding season in Great Tits, Parus major. We simulated neighbour and stranger intrusions at three breeding stages corresponding to key ecological and social situations, egg-laying, incubation and nestling stages, and examined the territorial songs and physical responses of territory owners. Our results suggest that neighbours are dear enemies in the egg-laying and incubation stages, when territories are stable, but not at the nestling stage when territories become unstable. In addition, physical responses were more effective as warning signals than territorial songs when dear enemy effects were expressed. We conclude that the dear enemy effect is expressed flexibly, rather than in a fixed way, by territorial male Great Tits in our study area.

High-frequency hearing is important for the survival of both echolocating bats and whales, but our understanding of its genetic basis is scattered and segmented. In this study, we combined RNA-Seq and comparative genomic analyses to obtain insights into the comprehensive gene expression profile of the cochlea and the adaptive evolution of hearing-related genes. A total of 144 genes were found to have been under positive selection in various species of echolocating bats and toothed whales, 34 of which were identified to be related to hearing behavior or auditory processes. Subsequently, multiple physiological processes associated with those genes were found to have adaptively evolved in echolocating bats and toothed whales, including cochlear bony development, antioxidant activity, ion balance, and homeostatic processes, along with signal transduction. In addition, abundant convergent/parallel genes and sites were detected between different pairs of echolocator species; however, no specific hearing-related physiological pathways were enriched by them and almost all of the convergent/parallel signals were selectively neutral, as previously reported. Notably, two adaptive parallel evolved sites in TECPR2 were shown to have been under positive selection, indicating their functional importance for the evolution of echolocation and high-frequency hearing in laryngeal echolocating bats. This study deepens our understanding of the genetic bases underlying high-frequency hearing in the cochlea of echolocating bats and toothed whales.

Background In genus Rhinolophus, species in the Rhinolophus philippinensis and R. macrotis groups are unique because the horseshoe bats in these group have relatively low echolocation frequencies and flight speeds compared with other horseshoe bats with similar body size. The different characteristics among bat species suggest particular evolutionary processes may have occurred in this genus. To study the adaptive evidence in the mitochondrial genomes (mitogenomes) of rhinolophids, especially the mitogenomes of the species with low echolocation frequencies, we sequenced eight mitogenomes and used them for comparative studies of molecular phylogeny and adaptive evolution. Results Phylogenetic analysis using whole mitogenome sequences produced robust results and provided phylogenetic signals that were better than those obtained using single genes. The results supported the recent establishment of the separate macrotis group. The signals of adaptive evolution discovered in the Rhinolophus species were tested for some of the codons in two genes (ND2 and ND6) that encode NADH dehydrogenases in oxidative phosphorylation system complex I. These genes have a background of widespread purifying selection. Signals of relaxed purifying selection and positive selection were found in ND2 and ND6, respectively, based on codon models and physicochemical profiles of amino acid replacements. However, no pronounced overlap was found for non-synonymous sites in the mitogenomes of all the species with low echolocation frequencies. A signal of positive selection for ND5 was found in the branch-site model when R. philippinensis was set as the foreground branch. Conclusions The mitogenomes provided robust phylogenetic signals that were much more informative than the signals obtained using single mitochondrial genes. Two mitochondrial genes that encoding proteins in the oxidative phosphorylation system showed some evidence of adaptive evolution in genus Rhinolophus and the positive selection signals were tested for ND5 in R. philippinensis. These results indicate that mitochondrial protein-coding genes were targets of adaptive evolution during the evolution of Rhinolophus species, which might have contributed to a diverse range of acoustic adaptations in this genus.

The bats skin microbiota plays an important role in reducing pathogen infection, including the deadly fungal pathogen Pseudogymnoascus destructans, the causative agent of white-nose syndrome. However, the dynamic of skin bacterial communities response to environmental perturbations remains poorly described. We characterized skin bacterial community over time and space in Rhinolophus ferrumequinum, a species with high resistance to the infection with P. destructans. We collected environmental covariate data to determine what factors influenced changes in community structure. We observed significant temporal and spatial shifts in the skin bacterial community, which was mainly associated with variation in operational taxonomic units. The skin bacterial community differed by the environmental microbial reservoirs and was most influenced by host body condition, bat roosting temperature and geographic distance between sites, but was not influenced by pathogen infection. Furthermore, the skin microbiota was enriched in particular taxa with antifungal abilities, such as Enterococcus, Burkholderia, Flavobacterium, Pseudomonas, Corynebacterium and Rhodococcus. And specific strains of Pseudomonas, Corynebacterium and Rhodococcus even inhibited P. destructans growth. Our findings provide new insights in characterizing the variation in bacterial communities can inform us about the processes of driving community assembly and predict the host's ability to resist or survive pathogen infection.

Many bat pups emit isolation calls with low-frequency components that facilitate maternal recognition. Most previous studies have noted that isolation calls generally have unique individual identit...

Bats are altricial, gregarious, and highly vocal animals. As such, they are compelling animal models for bioacoustic analyses of language-related traits. Insight into the vocal development of bats ...

Drift, selection, or their combined effects commonly drive geographic variation in traits. Clarifying the relative roles of each process is a long-standing research goal in evolutionary biology. Acoustic signals of bats are a phenotypic characteristic that plays an important role in social organization and species recognition. We extensively sampled the Chinese rufous horseshoe bat (Rhinolophus sinicus) throughout China and Vietnam and reconstructed a species phylogeny to better understand the patterns and causes of the geographic variation of acoustic signals. Our results showed that the resting frequency (RF) of calls varied with latitude, sex, and distance among different colony locations. RF differences were not correlated with genetic distance (based on only one nuclear locus and the mitochondrial locus), climatic factors (mean annual temperature and mean annual precipitable water), or body size, although differences in calls increased with distance among various populations. This suggests that cultural drift may play more important roles than genetic drift and acoustic adaptation in shaping acoustic differences within regions in R. sinicus.

Background Although the sensory drive hypothesis can explain the geographic variation in echolocation frequencies of some bat species, the molecular mechanisms underlying this phenomenon are still unclear. The three lineages of greater horseshoe bat (Rhinolophus ferrumequinum) in China (northeast, central-east, and southwest) have significant geographic variation in resting frequencies (RF) of echolocation calls. Because their cochleae have an acoustic fovea that is highly sensitive to a narrow range of frequencies, we reported the transcriptomes of cochleae collected from three genetic lineages of R. ferrumequinum, which is an ideal organism for studying geographic variation in echolocation signals, and tried to understand the mechanisms behind this bat phenomenon by analyzing gene expression and sequence variation. Results A total of 8190 differentially expressed genes (DEGs) were identified. We identified five modules from all DEGs that were significantly related to RF or forearm length (FL). DEGs in the RF-related modules were significantly enriched in the gene categories involved in neural activity, learning, and response to sound. DEGs in the FL-related modules were significantly enriched in the pathways related to muscle and actin functions. Using 21,945 single nucleotide polymorphisms, we identified 18 candidate unigenes associated with hearing, five of which were differentially expressed among the three populations. Additionally, the gene ERBB4, which regulates diverse cellular processes in the inner ear such as cell proliferation and differentiation, was in the largest module. We also found 49 unigenes that were under positive selection from 4105 one-to-one orthologous gene pairs between the three R. ferrumequinum lineages and three other Chiroptera species. Conclusions The variability of gene expression and sequence divergence at the molecular level might provide evidence that can help elucidate the genetic basis of geographic variation in echolocation signals of greater horseshoe bats.

Species delimitation and evolutionary reconstruction remain challenging for non-model species that have experienced reticulate evolution and exhibit conflicting patterns of differentiation among multiple lines of evidence, such as mitochondrial and nuclear data and phenotypes. Here, we applied an integrative taxonomic approach to a case study of Rhinolophus macrotis complex, whose taxonomic status remains controversial, to provide insight into the systematics and evolutionary history of these species. By integrating traditional genetic markers with different modes of inheritance, genome-wide SNPs as well as phenotypic characteristics, we clarified the presence of three closely related species, R. episcopus, R. siamensis, and R. osgoodi, within this complex, and proposed a new taxonomic treatment for R. osgoodi. Our results suggested that hybridization and introgression are the main causes of low mtDNA divergence in these species. Combined with the demographic inference, we deduced that glacial-interglacial cycles drove geographic isolation and secondary contacts of these species, then promoted hybridization and lineage fusion among them, finally resulting in a reticulate evolutionary pattern. Overall, our study highlights the importance of combining multiple types of data to delimit species, especially those with conserved morphology, and to reveal the sophisticated processes of speciation.

Determining the evolutionary history and population drivers, such as past large-scale climatic oscillations, stochastic processes and ecological adaptations, represents one of the aims of evolutionary biology. Hipposideros larvatus is a common bat species in Southern China, including Hainan Island. We examined genetic variation in H. larvatus using mitochondrial DNA and nuclear microsatellites. We found a population structure on both markers with a geographic pattern that corresponds well with the structure on mainland China and Hainan Island. To understand the contributions of geography, the environment and colonization history to the observed population structure, we tested isolation by distance (IBD), isolation by adaptation (IBA) and isolation by colonization (IBC) using serial Mantel tests and RDA analysis. The results showed significant impacts of IBD, IBA and IBC on neutral genetic variation, suggesting that genetic variation in H. larvatus is greatly affected by neutral processes, environmental adaptation and colonization history. This study enriches our understanding of the complex evolutionary forces that shape the distribution of genetic variation in bats.

Although evolutionary theory predicts that parents should discriminate between their own and others' offspring when levels of parental care are large, in some colonial breeding mammals, strong selective pressure would be expected to drive the evolution of mutual recognition between parents and offspring. Although less well studied than parental discrimination, in both birds and mammals, offspring discrimination of parents has also been documented. Nocturnal bats depend heavily on acoustic signals for communication, although little is known about whether bat pups can recognize their mothers based solely on maternal calls. We first investigated whether maternal calls of pomona leaf-nosed bats differ statistically among individuals. Then, we tested whether pups recognize their mothers' calls using a playback experiment. Echolocation pulses and directive calls of mother bats contain enough individual characteristics to allow for individual discrimination. Playback experiments showed that the pups responded with more vocalizations to playback of their own mother's versus another mother's directive calls, but not to playback of their own mother's versus another mother's echolocation pulses, suggesting that offspring can distinguish directive calls, but not echolocation pulses, of their own mother from those of other mothers. To our knowledge, this is the first study in which directive calls from bat mothers have been used to elicit vocalizations from pups and the first study to indicate that pups can recognize their mothers based solely on directive calls. These results advance our understanding of the function of acoustic signals in mother–offspring recognition and the adaptive benefit of such communication, particularly the conditions under which a pup's recognition of its mother is critical.

Bats are an appealing animal for investigating the biological basis and evolution of speech-related traits in wild mammal systems. We investigated vocal development in Horsfield's leaf-nosed bat pups, Hipposideros larvatus. We found that newborn bats emit many types of isolation calls, including downward frequency modulation (DFM), arched frequency modulation (AFM), upward frequency modulation (UFM), and others. The gradual decrease of Euclidean distance measurements revealed that pup isolation calls (DFM, AFM, UFM) gradually change into adult social calls. Furthermore, one of the isolation call types (UFM type) encoded an individual signature. By contrast, we report that early echolocation calls and isolation calls develop in parallel. As pups develop, early echolocation calls gradually mature and transform into adult-like echolocation calls. The interval and duration of echolocation calls decreases, while the peak frequency and bandwidth of frequency modulation increases.

Palaeoclimatic oscillations and different landscapes frequently result in complex population-level structure or the evolution of cryptic species. Elucidating the potential mechanisms is vital to understanding speciation events. However, such complex evolutionary patterns have rarely been reported in bats. In China, the Rhinolophus macrotis complex contains a large form and a small form, suggesting the existence of a cryptic bat species. Our field surveys found these two sibling species have a continuous and widespread distribution with partial sympatry. However, their evolutionary history has received little attention. Here, we used extensive sampling, morphological and acoustic data, as well as different genetic markers to investigate their evolutionary history. Genetic analyses revealed discordance between the mitochondrial and nuclear data. Mitochondrial data identified three reciprocally monophyletic lineages: one representing all small forms from Southwest China, and the other two containing all large forms from Central and Southeast China, respectively. The large form showed paraphyly with respect to the small form. However, clustering analyses of microsatellite and Chd1 gene sequences support two divergent clusters separating the large form and the small form. Moreover, morphological and acoustic analyses were consistent with nuclear data. This unusual pattern in the R. macrotis complex might be accounted for by palaeoclimatic oscillations, shared ancestral polymorphism and/or interspecific hybridization.

We studied the postnatal development of morphological and vocal features in a free-ranging population of the Asian particolored bat, Vespertilio sinensis. There were no significant differences in postnatal growth rates between males and females on the basis of morphological changes. Body mass and forearm length followed a linear pattern of growth until 28 days of age at mean growth rates of 0.38 g/day and 1.15 mm/day, respectively and thereafter increased slowly. The length of total epiphyseal gap of the fourth metacarpal–phalangeal joint initially showed a linear increase for up to 16 days, followed by a linear decrease until day 46 at a mean rate of 0.18 mm/day. When forearm length was used in combination with changes in total epiphyseal gap length, we derived reliable age estimation equations for the 1–46 day age range. Of the three nonlinear growth models (logistic, Gompertz, and von Bertalanffy), the von Bertalanffy and logistic equations provided the best fit to the empirical curves for body mass and forearm length, respectively. Studies of vocal development showed that infants could emit short calls as precursors of echolocation calls after birth. Isolation calls and precursors of echolocation calls were both characterized by multiple harmonics. We observed a systematic increase in the value of the starting frequency without a significant change in the terminal frequency over the 3 week period of development. In addition, the duration of isolation calls increased until day 4 and then decreased, whereas, the duration of precursors of echolocation calls decreased continuously to reach a stable level.

Based on mark-recapture data, we studied the postnatal growth of Marshall's horseshoe bat (Rhinolophus marshalli) in Hekou County, Yunnan Province, China. Our results detected no significant differences both in body mass and in forearm length between males and females at birth. On average, young bats were not agile fliers until 31 days of age. Body mass and forearm length of pups followed a linear pattern of growth until day 13, with mean growth rates of 0.17 g/day and 1.22 mm/day, respectively, and thereafter growth rates decreased. Length of the total epiphyseal gap of the fourth metacarpal-phalangeal joint showed a linear increase up to 10 days followed by a linear decrease until day 40 with a mean rate of 0.09 mm/day. Together, two equations permitted estimation of the age of R. marshalli pups between 1 and 40 days of age. Of the three nonlinear growth models (logistic, Gompertz, and von Bertalanffy), the logistic equation provided the best fit to the empirical curves for body mass and forearm length.

Based on mark-recapture data, we studied the postnatal development of morphological features and vocalization of the pomona leaf-nosed bat (Hipposideros pomona). Morphological changes indicated that body mass and length of forearm followed a linear pattern of growth until 13 days of age at mean growth rates of 0.14 g/day and 1.08 mm/day, respectively, and thereafter, growth rates slowed. The length of the total epiphyseal gap of the fourth metacarpal–phalangeal joint showed a linear increase for up to 10 days, followed by a linear decrease until day 40 at a mean rate of 0.09 mm/day. Together, two equations permitted estimation of the age of H. pomona pups between 1 and 40 days. The logistic equation provided the best fit to the empirical curves for body mass and length of forearm. Studies of vocal development showed that the precursors of echolocation calls were not emitted until day 7 after birth. As the pups grew, the dominant frequency (DF) of isolation calls increased and number of harmonics (NH) decreased, whereas the duration remained relatively stable. The DF and BFM2 (the bandwidth of the terminal frequency-modulated sweep from the second harmonic) of the early echolocation calls increased; however, the NH and duration decreased.

We quantified changes in body mass, forearm length, and the total length of the epiphyseal gap of the fourth metacarpal-phalangeal joint of the ashy leaf-nosed bat (Hipposideros cineraceus) based on mark-recapture data obtained in Fangkong Cave in Hekou, Yunnan Province, China. We used these data to develop empirical growth curves, to derive growth rates, to establish age-predictive equations, and to compare growth parameters based on three nonlinear growth models. Forearm length and body mass of pups followed a linear pattern of growth until day 17, with mean growth rates of 0.81 mm/day and 0.09 g/day, respectively and thereafter their growth rates increased more slowly. The length of the epiphyseal gap initially increased linearly up to day 13 and then decreased linearly at a mean rate of 0.07 mm/day until day 37. An equation for estimating age based on forearm length was valid when this dimension was ≤ 27.6 mm, whereas the equation based on the length of the epiphyseal gap was valid for forearm lengths...

There have been few studies of the structural and evolutionary characteristics of the mitochondrial control region (CR) in rhinolophids, yet this could have important consequences for the interpretation of phylogenetic relationships within this group. Here we sequenced and analyzed the CR of 37 individuals from 12 Rhinolophus species, including 2 species from GenBank. The length of the CR ranged from 1335 to 1514 bp, and the base composition was very similar among species. The CR of horseshoe bats, like that of other mammals, could be subdivided into a central conserved domain (CD) and two flanking variable domains, extended termination associated sequences (ETAS), and conserved sequence blocks (CSB). Besides the common conserved blocks (ETAS1, ETAS2, F-B boxes, CSB1, CSB2 and CSB3) found in 3 domains, an ETAS2-like and a CSB1-like element were also detected in the ETAS and CSB domains, respectively, in all individuals. Notwithstanding a short tandem repeat (11 or 13 bp) between CSB1 and CSB2 in all specimens, the base composition, copy number and arrays are all variable. A long tandem repeat (79 bp) was only identified in the ETAS domain in one individual of R. pusillus. Phylogenetic reconstructions based on the CR sequences indicated that the molecular phylogenetic relationships among some Rhinolophus species were inconsistent with the results of phenetic analyses, but similar to phylogenetic constructions using cytochrome b. An unidentified species R. sp and 3 species from the philippinensis-group that were clearly morphologically different comprised a monophyletic group, which could have resulted from morphological independent evolution.

Because of its complicated systematics, the bent-winged bat is one of the most frequently studied bat species groups. In China, two morphologically similar bent-winged bat species, Miniopterus fuliginosus and Miniopterus magnater were identified, but their distribution range and genetic differentiation are largely unexplored. In this study, we applied DNA bar codes and two other mitochondrial DNA genes including morphological parameters to determine the phylogeny, genetic differentiation, spatial distribution, and morphological difference of the M. fuliginosus and M. magnater sampled from China and one site in Vietnam. Mitochondrial DNA gene genealogies revealed two monophyletic lineages throughout the Tropic of Cancer. According to DNA bar code divergences, one is M. fuliginosus corresponding to the Chinese mainland and the other is M. magnater corresponding to tropical regions including Hainan and Guangdong provinces of China and Vietnam. Their most recent common ancestor was dated to the early stage of the Quaternary glacial period (ca. 2.26 million years ago [Ma] on the basis of D-loop data, and ca. 1.69–2.37 Ma according to ND2). A population expansion event was inferred for populations of M. fuliginosus at 0.14 Ma. The two species probably arose in separate Pleistocene refugia under different climate zones. They significantly differed in forearm length, maxillary third molar width, and greatest length of the skull.

Postnatal changes in flight development, wing shape and wing bone lengths of 56 marked neonate Hipposideros pomona were investigated under natural conditions in southwest China. Flight experiments showed that pups began to flutter with a short horizontal displacement at 10 days and first took flight at 19 days, with most achieving sustained flight at 1 month old. Analysis of covariance on wingspan, wing area, and the other seven wing characteristics between 'pre-flight' and 'post-volancy' periods supports the hypothesis that growth had one 'pre-flight' trajectory and a different 'post-volancy' trajectory in bats. Wingspan, handwing length and area, armwing length and area, and total wing area increased linearly until the age of first flight, after which the growth rates decreased (all P0.001). Wing loading declined linearly until day 19 before ultimately decreasing to adult levels (P0.001). Additionally, the relationship of different pairwise combinations of bony components composing span-wise length and chord-wise length was evaluated to test the hypothesis that compensatory growth of wing bones in H. pomona occurred in both 'pre-flight' and 'post-volancy' periods. The frequency of short-long and long-short pairs was significantly greater than that of short-short, long-long pairs in most pairs of bone elements in adults. The results indicate that a bone 'shorter than expected' would be compensated by a bone or bones 'longer than expected', suggesting compensatory growth in H. pomona. The pairwise comparisons conducted in adults were also performed in young bats during 'pre-flight' and 'post-volancy' periods, demonstrating that compensatory growth occurred throughout postnatal ontogeny.

Patterns of intraspecific geographic variation of signaling systems provide insight into the microevolutionary processes driving phenotypic divergence. The acoustic calls of bats are sensitive to diverse evolutionary forces, but processes that shape call variation are largely unexplored. In China, Rhinolophus ferrumequinum displays a diverse call frequency and inhabits a heterogeneous landscape, presenting an excellent opportunity for this kind of research. We quantified geographic variation in resting frequency (RF) of echolocation calls, estimated genetic structure and phylogeny of R. ferrumequinum populations, and combined this with climatic factors to test three hypotheses to explain acoustic variation: genetic drift, cultural drift, and local adaptation. Our results demonstrated significant regional divergence in frequency and phylogeny among the bat populations in China's northeast (NE), central-east (CE) and southwest (SW) regions. The CE region had higher frequencies than the NE and SW regions. Drivers of RF divergence were estimated in the entire range and just the CE/NE region (since these two regions form a clade). In both cases, RF divergence was not correlated with mtDNA or nDNA genetic distance, but was significantly correlated with geographic distance and mean annual temperature, indicating cultural drift and ecological selection pressures are likely important in shaping RF divergence among different regions in China.

The diet of the bat Rhinolophus ferrumequinum was studied at the Zhi’an Village of Ji’an City in China, from June to August 2004. The bats were trained in a laboratory (volume: 9x4x4 m3). Foraging strategies of the bat were observed at night and prey remains were collected and identified. The results showed that the diet consisted mainly of Lepidoptera in summer, including 11 families, more than 30 species of moths, such as Noctuidae (36.6% by number), Sphingidae (24.1%), Geometridae (13.4%) and Limacodidae (9.5%). The length of culled wings ranged from 10–40 mm (97.7%). Pearson correlation analysis showed that the bat R. ferrumequinum foraged their prey selectively, but not opportunistically. From field studies, two ways were observed in which the bats retrieved their prey including aerial hawking during peak active period of the insects and flycatching during the insects’ non-peak activity period. The bats never gleaned prey from the ground, though they appeared to be well able to detect fluttering moths on the ground.