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Studying the diversity of gut microbiota and antibiotic resistance genes (ARGs) in endangered migratory birds, as well as their responses to different environments, can reflect the health status of migratory birds and the ecological risk of ARG transmission. The globally endangered Scaly-sided merganser (Mergus squamatus) is a typical migratory bird, yet the diversity of its intestinal microbiota and ARGs has not been reported. In this study, fecal samples were collected from two major riparian foraging areas utilized by M. squamatus in the Changbai Mountains of China: the Lushui River and the Manjiang River. The gut microbiota and ARG composition were characterized using 16S rRNA amplicon and metagenomic sequencing. The results indicated that the relative abundance of gut microbiota in the two habitats differed significantly, but the core flora was similar. Moreover, the relative abundance of pathogenic bacteria was high, suggesting that M. squamatus faces a high risk of disease. A total of 382 ARGs were identified, which were primarily resistant to multidrug, tetracycline, macrolides-lincosamides-streptogramins, and mupirocin. MsbA, abcA, rpoB2, and tetT were the most abundant shared ARGs between the two sites. The findings demonstrate that habitat influences the diversity of gut microbiota and ARGs in the endangered migratory M. squamatus. This species could serve as a reservoir for pathogenic bacteria and ARGs. Therefore, monitoring the diversity of gut microbiota and ARGs in M. squamatus across different habitats is essential for a comprehensive assessment of its health status and the ecological risk of ARG transmission in the future.

BackgroundThe skin fungal communities of animals play a crucial role in maintaining host health and defending against pathogens. Because fungal infections can affect the skin microbiota of bats, gaining a comprehensive understanding of the characteristics of healthy bat skin fungal communities and the ecological processes driving them provides valuable insights into the interactions between pathogens and fungi.MethodsWe used Kruskal–Wallis tests and Permutational Multivariate Analysis of Variance (PERMANOVA) to clarify differences in skin fungal community structure among bat species. A Generalized Linear Model (GLM) based on a quasi-Poisson distribution and partial distance-based redundancy analysis (db-RDA) was performed to assess the influence of variables on skin fungal communities. Using community construction models to explore the ecological processes driving fungal community changes, t-tests and Wilcoxon tests were used to compare the alpha diversity and species abundance differences between the fungal structure on bat species’ skin and the environmental fungal pool.ResultsWe found significant differences in the composition and diversity of skin fungal communities among bat species influenced by temperature, sampling site, and body mass index. Trophic modes and skin fungal community complexity also varied among bat species. Null model and neutral model analysis demonstrated that deterministic processes dominated the assembly of skin fungal communities, with homogeneous selection as the predominant process. Skin fungal communities on bat species were impacted by the environmental fungal reservoir, and actively selected certain amplicon sequence variants (ASVs) from the environmental reservoir to adhere to the skin.ConclusionIn this study, we revealed the structure and the ecological process driving the skin fungal community across bat species in northern China. Overall, these results broaden our knowledge of skin fungal communities among bat species, which may be beneficial to potential strategies for the protection of bats in China.

Abstract Background Mesenchymal stem cells (MSCs) hold a great promise for cell-based therapy in the field of regenerative medicine. In this study, we aimed to evaluate the safety and efficacy of intravenous infusion of human umbilical cord-derived MSCs (HUC-MSCs) in patients with aging frailty. Methods In this randomized, double-blind, placebo-controlled trial, participants diagnosed with aging frailty were randomly assigned to receive intravenous administrations of HUC-MSCs or placebo. All of serious adverse events and AEs were monitored to evaluate the safety of treatment during the 6-month follow-up. The primary efficacy endpoint was alteration of physical component scores (PCS) of SF-36 qualities of life at 6 months. The secondary outcomes including physical performance tests and pro-inflammatory cytokines, were also observed and compared at each follow-up visits. All evaluations were performed at 1 week, 1, 2, 3 and 6 months following the first intravenous infusion of HUC-MSCs. Results In the MSCs group, significant improvements in PCS of SF-36 were observed from first post-treatment visit and sustained throughout the follow-up period, with greater changes compared to the placebo group (p = 0.042). EQ-VAS scores of MSCs group improved significantly at 2 month (p = 0.023) and continued until the end of the 6-month visit (p = 0.002) in comparison to the placebo group. The timed up and go (TUG) physical performance test revealed significant group difference and showed continual enhancements over 6 months (p

Abstract Background The majority of bat species have developed remarkable echolocation ability, especially for the laryngeally echolocating bats along with high-frequency hearing. Adaptive evolution has been widely detected for the cochleae in the laryngeally echolocating bats, however, limited understanding for the brain which is the central to echolocation signal processing in the auditory perception system, the laryngeally echolocating bats brain may also undergo adaptive changes. Result In order to uncover the molecular adaptations related with high-frequency hearing in the brain of laryngeally echolocating bats, the genes expressed in the brain of Rhinolophus ferrumequinum (CF bat) and Myotis pilosus (FM bat) were both detected and also compared. A total of 346,891 genes were detected and the signal transduction mechanisms were annotated by the most abundant genes, followed by the transcription. In hence, there were 3,088 DEGs were found between the two bat brains, with 1,426 highly expressed in the brain of R. ferrumequinum, which were significantly enriched in the neuron and neurodevelopmental processes. Moreover, we found a key candidate hearing gene, ADCY1, playing an important role in the R. ferrumequinum brain and undergoing adaptive evolution in CF bats. Conclusions Our study provides a new insight to the molecular bases of high-frequency hearing in two laryngeally echolocating bats brain and revealed different nervous system activities during auditory perception in the brain of CF bats.

Abstract Niche theory predicts that ecologically similar sympatric species should show differentiation in at least one of the main niche dimensions (time, space, and/or food). Here, we combined observations of breeding timing, nest site selection, and diet (the latter determined using DNA metabarcoding) to analyze the niche overlap and differentiation between two sympatric secondary cavity‐nesting birds, the Japanese Tit Parus minor and the Yellow‐rumped Flycatcher Ficedula zanthopygia. The results showed that (1) there were significant differences in the first egg laying date, length of the egg laying period, incubation date, and hatching date between tits and flycatchers, and the breeding time of flycatchers peaked later (about 30 days) than that of tits; (2) the two species had a large overlap in nest site selection, although the canopy coverage and shrub density of flycatchers were significantly higher than those of tits; and (3) the niche overlap in diet was minimal, with both species heavily relying on Lepidoptera (39.6% and 63.7% for tits and flycatchers, respectively), but with flycatchers consuming significantly higher percentages of Lepidoptera, Diptera, and Coleoptera than tits. The results indicate that these two sympatric secondary cavity‐nesting species have significant niche differentiation in breeding time and diet, but little differentiation in nest site selection.

ABSTRACTThe gut microbiome is a symbiotic microbial community associated with the host and plays multiple important roles in host physiology, nutrition, and health. A number of factors have been shown to influence the gut microbiome, among which diet is considered to be one of the most important; however, the relationship between diet composition and gut microbiota in wild mammals is still not well recognized. Herein, we characterized the gut microbiota of bats and examined the effects of diet, host taxa, body size, gender, elevation, and latitude on the gut microbiota. The cytochrome C oxidase subunit I (COI) gene and 16S rRNA gene amplicons were sequenced from the feces of eight insectivorous bat species in southern China, including Miniopterus fuliginosus, Aselliscus stoliczkanus, Myotis laniger, Rhinolophus episcopus, Rhinolophus osgoodi, Rhinolophus ferrumequinum, Rhinolophus affinis, and Rhinolophus pusillus. The results showed that the composition of gut microbiome and diet exhibited significant differences among bat species. Diet composition and gut microbiota were significantly correlated at the order, family, genus, and operational taxonomic unit levels, while certain insects had a marked effect on the gut microbiome at specific taxonomic levels. In addition, elevation, latitude, body weight of bats, and host species had significant effects on the gut microbiome, but phylosymbiosis between host phylogeny and gut microbiome was lacking. These findings clarify the relationship between gut microbiome and diet and contribute to improving our understanding of host ecology and the evolution of the gut microbiome in wild mammals.IMPORTANCEThe gut microbiome is critical for the adaptation of wildlife to the dynamic environment. Bats are the second-largest group of mammals with short intestinal tract, yet their gut microbiome is still poorly studied. Herein, we explored the relationships between gut microbiome and food composition, host taxa, body size, gender, elevation, and latitude. We found a significant association between diet composition and gut microbiome in insectivorous bats, with certain insect species having major impacts on gut microbiome. Factors like species taxa, body weight, elevation, and latitude also affected the gut microbiome, but we failed to detect phylosymbiosis between the host phylogeny and the gut microbiome. Overall, our study presents novel insights into how multiple factors shape the bat’s gut microbiome together and provides a study case on host-microbe interactions in wildlife.

Glued laminated timber stands as an increasingly favored engineered wood product in low-rise modern timber structures due to its commendable structural performance and carbon sequestration potential. Chinese standards presently fail to differentiate the effects of distinct wood species on the fire resistance of glulam. This study examined the charring rate and residual bearing capacity of glulam columns made from six commonly used wood species in China: poplar, Chinese fir, Douglas fir, hemlock, larch, and spruce. These materials exhibited densities ranging between 390 and 645 kg·m−³. ISO standard fire tests were conducted employing one- and two-adjacent-side fire exposures. Notably, the findings unveiled that high-density glulam columns demonstrated lower charring rates and slower rates of heat penetration. The two-adjacent-side fire configuration induced a corner effect, consequently amplifying the effective charring depth. The results showed a linear relationship between the ratio of diagonal to horizontal charring rate of the columns exposed to the two-adjacent-side fire and wood density. To enhance practical application, a formula was developed, employing the residual section method, to calculate the column's residual bearing capacity post two-adjacent-side fire exposure. This formula incorporates an adjustment depth to accommodate the corner effect, effectively mitigating the reduced wood strength within the pyrolysis zone.

ABSTRACT Skin acts as a mechanical barrier between the body and its surrounding environment and plays an important role in resistance to pathogens. However, we still know little regarding skin responses to physiological changes, particularly with regard to responses against potential pathogens. We herein executed RNA-seq on the wing of the Rhinolophus ferrumequinum to assess gene-expression variations at four physiological stages: pre-hibernation, hibernation (early-hibernation and late-hibernation), and post-hibernation, as well as the gene-expression patterns of infected and uninfected bats with the Pseudogymnoascus destructans (Pd). Our results showed that a greater number of differentially expressed genes between the more disparate physiological stages. Functional enrichment analysis showed that the down-regulated response pathways in hibernating bats included phosphorus metabolism and immune response, indicating metabolic suppression and decreased whole immune function. We also found up-regulated genes in post-hibernating bats that included C-type lectin receptor signalling, Toll-like receptor signalling pathway, and cell adhesion, suggesting that the immune response and skin integrity of the wing were improved after bats emerged from their hibernation and that this facilitated clearing Pd from the integument. Additionally, we found that the genes involved in cytokine or chemokine activity were up-regulated in late-hibernation compared to early-hibernation and that FOSB regulation of immune cell activation was differentially expressed in bats infected with Pd during late-hibernation, implying that the host’s innate immune function was enhanced during late-hibernation so as to resist pathogenic infection. Our findings highlight the concept that maintenance of intrinsic immunity provides protection against pathogenic infections in highly resistant bats.

Abstract Heterogeneous pathogenic stress can shape major histocompatibility complex (MHC) diversity by influencing the functional plasticity of the immune response. Therefore, MHC diversity could reflect environmental stress, demonstrating its importance in uncovering the mechanisms of adaptive genetic variation. In this study, we combined neutral microsatellite loci, an immune‐related MHC II‐DRB locus, and climatic factors to unravel the mechanisms affecting the diversity and genetic differentiation of MHC genes in the greater horseshoe bat (Rhinolophus ferrumequinum), a species with a wide geographical distribution that has three distinct genetic lineages in China. First, increased genetic differentiation at the MHC locus among populations compared using microsatellites indicated diversifying selection. Second, the genetic differentiation of MHC and microsatellites were significantly correlated, suggesting that demographic processes exist. However, MHC genetic differentiation was significantly correlated with geographical distance among populations, even after controlling for the neutral markers, suggesting a major effect of selection. Third, although the MHC genetic differentiation was larger than that for microsatellites, there was no significant difference in the genetic differentiation between the two markers among genetic lineages, indicating the effect of balancing selection. Fourth, combined with climatic factors, MHC diversity and supertypes showed significant correlations with temperature and precipitation, but not with the phylogeographic structure of R. ferrumequinum, suggesting an effect of local adaptation driven by climate on MHC diversity. Moreover, the number of MHC supertypes varied between populations and lineages, suggesting regional characteristics and support for local adaptation. Taken together, the results of our study provide insights into the adaptive evolutionary driving forces at different geographic scales in R. ferrumequinum. In addition, climate factors may have played a vital role in driving adaptive evolution in this species.

Arylalkylamine N-acetyltransferase (AANAT) is a crucial rate-limiting enzyme in the synthesis of melatonin. AANAT has been confirmed to be independently duplicated and inactivated in different animal taxa in order to adapt to the environment. However, the evolutionary forces associated with having a single copy of AANAT remain unclear. The greater horseshoe bat has a single copy of AANAT but exhibits different hibernation rhythms in various populations. We analyzed the adaptive evolution at the gene and protein levels of AANAT from three distinct genetic lineages in China: northeast (NE), central east (CE), and southwest (SW). The results revealed greater genetic diversity in the AANAT loci of the NE and CE lineage populations that have longer hibernation times, and there were two positive selection loci. The catalytic capacity of AANAT in the Liaoning population that underwent positive selection was significantly higher than that of the Yunnan population (p < 0.05). This difference may be related to the lower proportion of α helix and the variation in two interface residues. The adaptive evolution of AANAT was significantly correlated with climate and environment (p < 0.05). After controlling for geographical factors (latitude and altitude), the evolution of AANAT by the negative temperature factor was represented by the monthly mean temperature (r = −0.6, p < 0.05). The results identified the gene level variation, functional adaptation, and evolutionary driving factors of AANAT, provide an important foundation for further understanding the adaptive evolution of the single copy of AANAT in pteropods, and may offer evidence for adaptive hibernation rhythms in bats.

The mechanical characteristics of wood members in a fire are reduced as a result of strength loss of the outermost charring layer and strength reduction of the secondary outer layer in high temperature zones. The charring layer has strong oxygen isolation effects. The main mechanical characteristics of larch wood, including modulus of elasticity (MOE), parallel-to-grain compressive (fc), tensile (ft), bending strengths (fb), and ductility coefficient, were determined under oxygen-free conditions at higher temperatures. Fourier transform-infrared (FT-IR) and scanning electron microscopies (SEM) were used to analyze chemical composition and microscopic changes of wood. The results suggested that hemicellulose softening resulted in decreases in mechanical properties of wood between 20 and 70 °C. From 70 to 190 °C, FT-IR spectroscopy provided evidence that increases in MOE and fc, with relatively minor changes in fb and ft, were associated with cellulose crystallinity and lignin cross-linking/condensation. The use of SEM further indicated degradation of wood composition.

Understanding trophic niche differentiation is critical for exploring interspecific competition and stable coexistence among morphologically similar sympatric species. Bats are an ideal model for studying trophic niche differentiation among species because of their high taxonomic and ecological diversities, as well as their special life history traits. Although many factors can affect bat trophic niches, few studies have combined multiple factors to investigate the influences on bat trophic niches. In this study, we analyzed the summer diet and potential influencing factors of five sympatric rhinolophid bats in southwestern China: Rhinolophus macrotis, Rhinolophus osgoodi, Rhinolophus ferrumequinum, Rhinolophus affinis, and Rhinolophus pusillus. All five species mainly fed on Lepidopteran and Dipteran insects, with a large trophic niche breadth for each species and a low degree of dietary overlap. With the exception of R. affinis and R. pusillus, significant differences in diet composition were detected among species, which indicated relatively low interspecific competition. Canonical correspondence analysis showed that both echolocation calls and body size significantly affected interspecific diet differentiation, while wing morphology and bite force had relatively weak effects. This study suggests that the combined effects of multiple factors may drive trophic niche differentiation among five rhinolophid bat species in the study area.

Summary 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 μg 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.

Abstract 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.

Abstract In many vertebrates, vocal communication is crucial in parent–offspring interactions, and parents are often able to discriminate between the calls of their own and others' offspring. There are many reports on the unidirectional recognition of isolation calls of pups by maternal bats, but few studies on the ability of bat pups to recognize maternal acoustic signals. In this study, we investigated whether the echolocation pulses of female Asian particolored bats (Vespertilio sinensis) and isolation calls of pups differ statistically among individuals. We used two‐choice playback experiments to test whether the mothers and pups of V. sinensis can recognize each other by acoustic signals. Both the echolocation pulses of mother bats and the isolation calls of pups contained sufficient individual characteristics. Playback experiments showed that mothers were able to recognize isolation calls of pups, and most pups greater than 12 days old were able to distinguish echolocation pulses of their own mother from those of other mothers. This is the first use of two‐choice acoustic signal playback experiments to confirm that pups can recognize their mothers by echolocation calls. The results provide behavioral evidence for bidirectional recognition of acoustic signals between mothers and infants in frequency‐modulated type bats.

The genus Myotis is one of the most diverse and widely distributed mammals, providing a good model for studies of speciation and diversification across large geographic scales. However, the classification within this genus has long been chaotic. Taxonomic revisions based on multiple data sources are essential and urgent. In this study, morphometrics and genetic markers with different modes of inheritance were used to clarify the taxonomy of Myotis distributed in China. Based on 173 mitochondrial Cytb sequences and five morphological characteristics, 114 specimens collected nationwide over the past 20 years were assigned to 11 Myotis species. All Chinese samples classified into M. davidii and M. longipes were revised to M. alticraniatus and M. laniger. Then, two nuclear fragments (Rag2 and Chd1) and Cytb sequences from representative individuals of Chinese Myotis were used for multilocus phylogeny reconstruction and genetic divergence evaluation. The phylogenetic relationships were clearly demonstrated in the species tree: M. alticraniatus and M. laniger; M. fimbriatus, M. pilosus, M. macrodactylus, and M. petax; and M. pequinius, M. chinensis, and M. blythii formed three strongly supported monophyletic clades. Mitochondrial divergence was almost 10 times that of nuclear divergence, with interspecific K2P distances ranging from 8% to 20% for Cytb and 0.3% to 2.3% for concatenated nuclear genes. Low levels of genetic divergence were observed between M. alticraniatus and M. laniger, as well as M. fimbriatus and M. pilosus. These results provide new insights into the taxonomy and phylogeny of Myotis bats in China and are important for the future research and conservation of Chinese Myotis.

In order to unravel the effect of aerated irrigation on soil dissolved organic matter (DOM) fluorescence characteristics, and humification degree, a randomized block experiment was conducted with three factors and a two-level design, i.e., two irrigation rates (0.6 and 1.0 times of crop evaporation pan coefficient, W1 and W2), two nitrogen application rates (225 and 300 kg hm−2, N1 and N2), and two aeration rates (15% and 0% in control treatment, A1 and A0). Fluorescence regional integration (FRI) and correlation analysis methods were used to investigate the evolution characteristics of the soil DOM fluorescence spectrum. Under aerated and conventional subsurface irrigation, soil DOM components were dominated by humic acid-like substances, fulvic acid-like substances, tryptophan-like proteins, and supplemented by tyrosine-like proteins and dissolved microbial metabolites. Soil aeration could promote the consumption of soil DOM components under low irrigation rates and accelerate the consumption of soil DOM components under high irrigation rates. The humification index of AI treatments varied from 8.47 to 9.94 during the maturity growth stage of pepper, averagely increased by 31.59% compared with the non-aeration treatment. To sum up, aerated irrigation can promote the depletion of small molecular proteins and accelerate nutrient turnover and the accumulation of big molecular proteins.

Host-associated skin bacteria are essential for resisting pathogen infections and maintaining health. However, we have little understanding of how chiropteran skin microbiota are distributed among bat species and their habitats, or of their putative roles in defending against Pseudogymnoascus destructans in China. In this study, we characterized the skin microbiomes of four bat species at five localities using 16S rRNA gene amplicon sequencing to understand their skin microbial composition, structure, and putative relationship with disease. The alpha- and beta-diversities of skin microbiota differed significantly among the bat species, and the differences were affected by environmental temperature, sampling sites, and host body condition. The chiropteran skin microbial communities were enriched in bacterial taxa that had low relative abundances in the environment. Most of the potential functions of skin microbiota in bat species were associated with metabolism. Focusing on their functions of defense against pathogens, we found that skin microbiota could metabolize a variety of active substances that could be potentially used to fight P. destructans. The skin microbial communities of bats in China are related to the environment and the bat host, and may be involved in the host’s defense against pathogens.

Chytridiomycosis is an amphibian fungal disease caused by Batrachochytrium dendrobatidis (Bd), which has caused large-scale death and population declines on several continents around the world. To determine the current status of Bd infection in amphibians, we conducted a global meta-analysis. Using PubMed, ScienceDirect, SpringerLink, China National Knowledge Infrastructure (CNKI) and Wanfang database searches, we retrieved a total of 111 articles from 2000 to 2021. Based on these, we estimated the Bd prevalence to be 18.54% (95% CI: 13.76–20.52) in current extent amphibians. Among these populations, the prevalence of Bd in Asia was the lowest at 7.88% (95% CI: 1.92–8.71). Further, no Bd infection was found in Vietnam. However, the prevalence of Bd in Oceania was the highest at 36.34% (95% CI: 11.31–46.52). The Bd prevalence in Venezuela was as high as 49.77% (95% CI: 45.92–53.62). After 2009, the global Bd prevalence decreased to 18.91% (95% CI: 13.23–21.56). The prevalence of Bd in epizootic populations was significantly higher than enzootic populations. The highest prevalence of Bd was detected with real-time PCR at 20.11% (95% CI: 13.12–21.38). The prevalence of Bd in frogs was the highest at 20.04% (95% CI: 13.52–21.71), and this different host was statistically significant (P < 0.05). At the same time, we analyzed the geographic factors (longitude, latitude, elevation, rainfall and temperature) that impacted the fungal prevalence in amphibians. Our meta-analysis revealed that factors including region, disease dynamic, detection method, host and climate may be sources of the observed heterogeneity. These results indicate that chytridiomycosis was a consistent threat to amphibians from 2000 to 2021. Based on different habitat types and geographical conditions, we recommend formulating corresponding control plans and adopting reasonable and efficient biological or chemical methods to reduce the severity of such diseases.

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.

Rickett’s big-footed bat, Myotis pilosus, which belongs to the family Vespertilionida, is the only known piscivorous bat in East Asia. Accurate whole genome and transcriptome annotations are essential for the study of bat biological evolution. The lack of a whole genome for M. pilosus has limited our understanding of the molecular mechanisms underlying the species’ evolution, echolocation, and immune response. In the present work, we sequenced the entire transcriptome using error-corrected PacBio single-molecule real-time (SMRT) data. Then, a total of 40 GB of subreads were generated, including 29,991 full-length non-chimeric (FLNC) sequences. After correction by Illumina short reads and de-redundancy, we obtained 26,717 error-corrected isoforms with an average length of 3018.91 bp and an N50 length of 3447 bp. A total of 1528 alternative splicing (AS) events were detected by transcriptome structural analysis. Furthermore, 1032 putative transcription factors (TFs) were identified, with additional identification of several long non-coding RNAs (lncRNAs) with high confidence. Moreover, several key genes, including PRL-2, DPP4, Glul, and ND1 were also identified as being associated with metabolism, immunity, nervous system processes, and auditory perception. A multitude of pattern recognition receptors was identified, including NLR, RLR, SRCR, the antiviral molecule IRF3, and the IFN receptor subunit IFNAR1. High-quality reference genomes at the transcriptome level may be used to quantify gene or transcript expression, evaluate alternative splicing levels, identify novel transcripts, and enhance genome annotation in bats.

In the process of species differentiation and adaption, the relative influence of natural selection on gene expression variation often remains unclear (especially its impact on phenotypic divergence). In this study, we used differentially expressed genes from brain, cochlea, and liver samples collected from two species of bats to determine the gene expression variation forced by natural selection when comparing at the interspecific (Rhinolophus siamensis and R. episcopus episcopus) and the intraspecific (R. e. episcopus and R. episcopus spp.) levels. In both cases, gene expression variation was extensively adaptive (>66.0%) and mainly governed by directional selection, followed by stabilizing selection, and finally balancing selection. The expression variation related to acoustic signals (resting frequency, RF) and body size (forearm length, FA) was also widely governed by natural selection (>69.1%). Different functional patterns of RF- or FA-related adaptive expression variation were found between the two comparisons, which manifested as abundant immune-related regulations between subspecies (indicating a relationship between immune response and phenotypic adaption). Our study verifies the extensive adaptive expression variation between both species and subspecies and provides insight into the effects of natural selection on species differentiation and adaptation as well as phenotypic divergence at the expression level.

Abstract 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.

AbstractHigh-frequency hearing is particularly important for echolocating bats and toothed whales. Previously, studies of the hearing-related genes PrestinKCNQ4TMC1ShhSK2ShhSK2ShhSK2ShhSK2SK2SK2

Background The gut microbiota is closely linked to host development, diet and health and is influenced by both the host and the environment. Although many studies have focused on the dynamics of the gut microbiota during development in captive animals, few studies have focused on the dynamics of the gut microbiota during development in wild animals, especially for the order Chiroptera. Methods In this study, we characterized the gut microbiota of the wild Asian particolored bat (Vespertilio sinensis) from 1 day to 6 weeks after birth. We explored the changes in their gut microbial community compositions, examined possible influencing factors, and predicted the feeding transition period. Results The gut microbiota changed during the development of V. sinensis. The alpha diversity of the bats’ gut microbiota gradually increased but did not change significantly from the 1st day to the 4th week after birth; however, the alpha diversity decreased significantly in week 5, then stabilized. The beta diversity differed slightly in weeks 4–6. In week 4, the fecal samples showed the highest diversity in bacterial community composition. Thus, we predicted that the potential feeding transition period for V. sinensis may occur during week 4. Redundancy analysis showed that age and body mass index significantly affected the compositional changes of the gut microbiota in Asian particolored bats. Conclusion The gut microbiota changed during the development of V. sinensis. We suggest that changes in the alpha and beta diversity during week 4 after birth indicate a potential feeding transition, highlighting the importance of diet in the gut microbiota during the development of V. sinensis.

Background Diet plays a crucial role in sculpting microbial communities. Similar diets appear to drive convergence of gut microbial communities between host species. Captivity usually provides an identical diet and environment to different animal species that normally have similar diets. Whether different species’ microbial gut communities can be homogenized by a uniform diet in captivity remains unclear. Methods In this study, we compared gut microbial communities of three insectivorous bat species (Rhinolophus ferrumequinum, Vespertilio sinensis, and Hipposideros armiger) in captivity and in the wild using 16S rDNA sequencing. In captivity, R. ferrumequinum and V. sinensis were fed yellow mealworms, while H. armiger was fed giant mealworms to rule out the impact of an identical environment on the species’ gut microbial communities. Results We found that the microbial communities of the bat species we studied clustered by species in the wild, while the microbial communities of R. ferrumequinum and V. sinensis in captivity clustered together. All microbial functions found in captive V. sinensis were shared by R. ferrumequinum. Moreover, the relative abundances of all metabolism related KEGG pathways did not significantly differ between captive R. ferrumequinum and V. sinensis; however, the relative abundance of “Glycan Biosynthesis and Metabolism” differed significantly between wild R. ferrumequinum and V. sinensis. Conclusion Our results suggest that consuming identical diets while in captivity tends to homogenize the gut microbial communities among bat species. This study further highlights the importance of diet in shaping animal gut microbiotas.

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.

ABSTRACT Globalization has facilitated the worldwide movement and introduction of pathogens, but epizoological reconstructions of these invasions are often hindered by limited sampling and insufficient genetic resolution among isolates. Pseudogymnoascus destructans, a fungal pathogen causing the epizootic of white-nose syndrome in North American bats, has exhibited few genetic polymorphisms in previous studies, presenting challenges for both epizoological tracking of the spread of this fungus and for determining its evolutionary history. We used single nucleotide polymorphisms (SNPs) from whole-genome sequencing and microsatellites to construct high-resolution phylogenies of P. destructans. Shallow genetic diversity and the lack of geographic structuring among North American isolates support a recent introduction followed by expansion via clonal reproduction across the epizootic zone. Moreover, the genetic relationships of isolates within North America suggest widespread mixing and long-distance movement of the fungus. Genetic diversity among isolates of P. destructans from Europe was substantially higher than in those from North America. However, genetic distance between the North American isolates and any given European isolate was similar to the distance between the individual European isolates. In contrast, the isolates we examined from Asia were highly divergent from both European and North American isolates. Although the definitive source for introduction of the North American population has not been conclusively identified, our data support the origin of the North American invasion by P. destructans from Europe rather than Asia. IMPORTANCE This phylogenetic study of the bat white-nose syndrome agent, P. destructans, uses genomics to elucidate evolutionary relationships among populations of the fungal pathogen to understand the epizoology of this biological invasion. We analyze hypervariable and abundant genetic characters (microsatellites and genomic SNPs, respectively) to reveal previously uncharacterized diversity among populations of the pathogen from North America and Eurasia. We present new evidence supporting recent introduction of the fungus to North America from a diverse Eurasian population, with limited increase in genetic variation in North America since that introduction.

Genetic diversity is one of the pillars of conservation biology research. High genetic diversity and abundant genetic variation in an organism may be suggestive of capacity to adapt to various environmental changes. The major histocompatibility complex (MHC) is known to be highly polymorphic and plays an important role in immune function. It is also considered an ideal model system to investigate genetic diversity in wildlife populations. The Rufous-backed Bunting (Emberiza jankowskii) is an endangered species that has experienced a sharp decline in both population and habitat size. Many historically significant populations are no longer present in previously populated regions, with only three breeding populations present in Inner Mongolia (i.e., the Aolunhua, Gahaitu and Lubei557 populations). Efforts focused on facilitating the conservation of the Rufous-backed Bunting (Emberiza jankowskii) are becoming increasingly important. However, the genetic diversity of E. jankowskii has not been investigated. In the present study, polymorphism in exon 2 of the MHCIIB of E. jankowskii was investigated. This polymorphism was subsequently compared with a related species, the Meadow Bunting (Emberiza cioides). A total of 1.59 alleles/individual were detected in E. jankowskii and 1.73 alleles/individual were identified in E. cioides. The maximum number of alleles per individual from the three E. jankowskii populations suggest the existence of at least three functional loci, while the maximum number of alleles per individual from the three E. cioides populations suggest the presence of at least four functional loci. Two of the alleles were shared between the E. jankowskii and E. cioides. Among the 12 unique alleles identified in E. jankowskii, 10.17 segregating sites per allele were detected, and the nucleotide diversity was 0.1865. Among the 17 unique alleles identified in E. cioides, eight segregating sites per allele were detected, and the nucleotide diversity was 0.1667. Overall, compared to other passerine birds, a relatively low level of MHC polymorphism was revealed in E. jankowskii, which was similar to that in E. cioides. Positive selection was detected by PAML/SLAC/FEL analyses in the region encoding the peptide-binding region in both species, and no recombination was detected. Phylogenetic analysis showed that the alleles from E. jankowskii and E. cioides belong to the same clade and the two species shared similar alleles, suggesting the occurrence of a trans-species polymorphism between the two Emberiza species.

The greater horseshoe bat, Rhinolophus ferrumequinum, is an important model organism for studies on chiropteran phylogeographic patterns. Previous studies revealed the population history of R. ferrumequinum from Europe and most Asian regions, yet there continue to be arguments about their evolutionary process in Northeast Asia. In this study, we obtained mitochondrial DNA cyt b and D-loop data of R. ferrumequinum from Northeast China, South Korea and Japan to clarify their phylogenetic relationships and evolutionary process. Our results indicate a highly supported monophyletic group of Northeast Asian greater horseshoe bats, in which Japanese populations formed a single clade and clustered into the mixed branches of Northeast Chinese and South Korean populations. We infer that R. ferrumequinum in Northeast Asia originated in Northeast China and South Korea during a cold glacial period, while some ancestors likely arrived in Japan by flying or land bridge and subsequently adapted to the local environment. Consequently, during the warm Eemian interglaciation, the Korea Strait, between Japan and South Korea, became a geographical barrier to Japanese and inland populations, while the Changbai Mountains, between China and North Korea, did not play a significant role as a barrier between Northeast China and South Korea populations.

Hibernation is one type of torpor, a hypometabolic state in heterothermic mammals, which can be used as an energy-conservation strategy in response to harsh environments, e.g. limited food resource. The liver, in particular, plays a crucial role in adaptive metabolic adjustment during hibernation. Studies on ground squirrels and bears reveal that many genes involved in metabolism are differentially expressed during hibernation. Especially, the genes involved in carbohydrate catabolism are down-regulated during hibernation, while genes responsible for lipid β-oxidation are up-regulated. However, there is little transcriptional evidence to suggest physiological changes to the liver during hibernation in the greater horseshoe bat, a representative heterothermic bat. In this study, we explored the transcriptional changes in the livers of active and torpid greater horseshoe bats using the Illumina HiSeq 2000 platform. A total of 1358 genes were identified as differentially expressed during torpor. In the functional analyses, differentially expressed genes were mainly involved in metabolic depression, shifts in the fuel utilization, immune function and response to stresses. Our findings provide a comprehensive evidence of differential gene expression in the livers of greater horseshoe bats during active and torpid states and highlight potential evidence for physiological adaptations that occur in the liver during hibernation.

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.

Aerated irrigation (AI) has the potential to be an effective management strategy to further increase yield and nitrogen-use efficiency. To determine the correlation between aerated irrigation and nitrogen application, we conducted a greenhouse experiment in Zhengzhou, Henan, China. A two-factor, three- level experiment was set up in which three aeration rates of dissolved oxygen in irrigation water (A1 = 5 mg/L, A2 = 15 mg/L, and A3 = 40 mg/L) and three nitrogen application rates (N1 = 120 kg/hm², N2 = 180 kg/hm², and N3 = 240 kg/ hm²) were set by using randomized block testing. CK is normal subsurface irrigation without fertilization and aeration, seemed as a control treatment. The analysis of variance, correlation analysis, and structural equation model (SEM) clarify how each factor affects the yield and nitrogen absorption and the utilization of greenhouse tomatoes. (Results) The results show that aerated irrigation significantly improves the nitrogen uptake efficiency and tomato yield (p < 0.05). Compared with the non-aerated treatment, the tomato yield after aerated irrigation increased by 13.35%-29.73%, and the highest tomato yield was 250.42t/hm² under treatment A2N3. Compared with treatments A1N3 and A3N3, the yield of treatment A2N3 increased by 29.73% and 11.53%, respectively, and the tomato nitrogen absorption efficiency increased by 53.06% and 18.33%, respectively (p < 0.05). Under the action of a single factor, applying nitrogen under aerated conditions can increase tomato yield and nitrogen absorption. According to SEM analysis, aeration and nitrogen application are positively correlated to a significant extent with tomato nitrogen absorption and indirectly correlated positively with tomato yield; the path coefficients are 0.969 and 0.927, respectively. Therefore, the combination of aeration and nitrogen application can further increase yield and nitrogen absorption efficiency, and the aeration amount A2 (15 mg/L) and the nitrogen application rate N3 (240 kg/hm²) are the best combination to increase the nitrogen uptake and yield of tomatoes. The results of this research provide technical support for optimizing gas-nitrogen coupled drip-irrigation and should help to popularize the use of gas-nitrogen coupled irrigation. [ABSTRACT FROM AUTHOR]