Context-dependent responses of bats to artificial light at night in urban and natural woodlands.

Asian Rhinolophus bats are considered the natural reservoirs of an ancestral SARS-CoV-2. However, the biology of SARS-CoV-2-related viruses in bat cells is not well understood. Here, we investigated the replication of an isolate of BANAL-236, the only bat-derived SARS-CoV-2 relative isolated to date, in Rhinolophus ferrumequinum lungs (Rfe) cells. BANAL-236 did not replicate in wild-type Rhinolophus cell lines. Entry assays using pseudoviruses expressing the spike proteins (S) of SARS-CoV-2, BANAL-236, and BANAL-52 revealed that efficient S-mediated entry depends on the expression of human ACE2 (hACE2) and human TMPRSS2 (hTMPRSS2) in human and Rhinolophus cells. Through biochemical, virological, and electron microscopy analyses, we showed that BANAL-236 and SARS-CoV-2 completed their replication cycles in RFe cells engineered to express high levels of hACE2 and hTMPRSS2. Despite efficient viral replication in modified Rhinolophus and human cells, no induction of interferon (IFN)-stimulated genes was detected. Using a screening approach, we identified several BANAL-236 proteins that antagonize IFN production and signalling in human cells. Our findings thus show that BANAL-236 possesses critical features that enabled zoonotic spillover: hACE2 usage and potent evasion of human IFN responses. The Rhinolophus cellular model we established offers a platform for further investigating the interactions between bat sarbecoviruses and their reservoir hosts.

The greater horseshoe bat (Rhinolophus ferrumequinum), which is widely distributed across the temperate regions of China, primarily consists of two major evolutionary lineages: a northeastern (NE) lineage with a hibernation period of 6-8 months and a central-eastern (CE) lineage with a hibernation period of 4-5 months. This study conducted a comparative analysis of liver transcriptomes from these two lineages during the active, torpor, and arousal phases. The results indicated that the CE lineage exhibited a significantly greater number of differentially expressed genes (DEGs) compared to the NE lineage. During the torpor phase, both lineages transitioned from carbohydrate metabolism to lipid metabolism, substantially downregulating genes and pathways associated with amino acid metabolism, and upregulating immune-related genes to maintain essential defense functions. In the arousal phase, both lineages only moderately activated several genes associated with immunity and metabolic regulation to facilitate a rapid return to torpor. Notably, the number of DEGs co-regulated between the two lineages was very limited, and a large number of lineage-specific regulatory genes related to energy and metabolism were identified. This may reflect the adaptability of different bat lineages to the local environment, highlighting the importance of habitat conditions in lineage differentiation. Therefore, hibernation induces substantial transcriptomic reorganization in the liver of R. ferrumequinum, particularly affecting metabolic and immune processes. Distinct geographic lineages exhibit unique hibernation adaptation strategies through the regulation of specific genes and pathways. This study enhances the understanding of the molecular mechanisms underlying hibernation adaptation across different evolutionary lineages of the same species at the transcriptomic level, providing insights into the evolutionary adaptations of animals to environmental changes.

Interactions between two protected species: Rhinolophus ferrumequinum impacting the recovery of Rhinolophus hipposideros populations in Britain

Locomotion and fluid pumping near surfaces are ubiquitous in nature, ranging from the slow crawling of snails to the rapid flight of bats. This study categorizes these behaviors based on the undulation number (Un) and Reynolds number ($Re$). We contrast low $Re$ undulatory propulsion ($\text{Un} \gt 1$), exemplified by freshwater snails, with high $Re$ flapping propulsion ($\text{Un} \lt 1$), seen in bats and bees. For snails, we derive lubrication models showing that pumping and swimming speeds scale with $(a/h_0)^2$, a result validated by robotic experiments which also reveal the detrimental effects of surface deformation (high capillary/Bond ratio). Conversely, for high $Re$ fliers, we examine the ground effect's role in lift enhancement. Biological data from bats (Hipposiderospratti and Rhinolophus ferrumequinum) reveal a 2.5-fold increase in lift coefficient during surface-skimming drinking flights, attributed to aerodynamic squeezing effects. Finally, we analyze honeybee fanning, demonstrating how a "jet-vortex" mechanism utilizes ground effect to transport pheromones efficiently against diffusion. These findings provide a unified framework for understanding fluid-structure interactions near boundaries in biological systems.

Abstract Mammalian torpor imposes unique metabolic constraints, yet the mechanisms of nitrogen metabolism during this state remain unclear. In this study, we show that the urea cycle is selectively regulated rather than broadly suppressed in torpid bats. A significantly increased abundance of carbamoyl-phosphate synthase 1 (CPS1) maintained its functional capacity during torpor and arousal in Myotis ricketti. Proteomic analyses and confocal microscopy identified a specific association and co-localization between CPS1 and agmatinase (AGMAT), an ATP-independent enzyme involved in nitrogen metabolism. Co-localization of CPS1 and AGMAT was also observed in torpid Rhinolophus ferrumequinum, a phylogenetically distant species. Fluorescence resonance energy transfer (FRET) further supported an indirect CPS1-AGMAT interaction. Most urea cycle enzymes exhibited stable or only moderately reduced expression during bat torpor, and metabolic profiling demonstrated sustained nitrogen flux. Together, these findings reveal a conserved adaptive mechanism that maintains urea cycle function, potentially enhancing osmotic stability and energy efficiency during prolonged fasting and water scarcity.

Bats represent the second largest order of mammals and are the only flying mammals, exhibiting remarkable biological characteristics such as viral tolerance, longevity, and low tumor incidence. Although bats serve as reservoirs for numerous zoonotic viruses, they remain asymptomatic without overt infection. However, the mechanisms and effects of B-cell adaptive immune responses in bats are largely unknown. In mammals, BCR V(D)J gene recombination and the formation of the CDR3 repertoire are key mechanisms underlying B-cell diversity, specificity, and memory responses. In this study, we performed, for the first time, a chromosome-level annotation of the IGK locus (17 KV; 4 KJ;1 LC) and IGL locus (74 LV; 6 LJ-LC gene clusters) in Rhinolophus ferrumequinum, as well as the IGH locus (81 HV; 16 HD; 6 HJ;HC of IgM, IgG, IgE, IgA) and IGL locus (56 LV; 9 LJ-LC gene clusters) in Antrozous pallidus. Comparative evolutionary analyses of each IG locus were also conducted. Using the annotated IG H/K/L constant region gene sequences, we employed 5′ Rapid Amplification of cDNA Ends (5′RACE) to construct BCR CDR3 repertoires and perform high-throughput sequencing (HTS). The annotated bat IG H/K/L genes were then used to build a MixCR tool for bat BCR CDR3 HTS analysis. In the spleen and intestinal tissues of bats from Hipposideros armige and Rhinolophus pearsonii/pusillus, we observed high diversity and differential preferential usage of V and J genes in the BCR CDR3 repertoires. Compared with human and mouse BCR CDR3 repertoires, bat CDR3 repertoires exhibited both similarities (e.g., high-frequency motifs, tyrosine enrichment) and distinct differences (e.g., N/P nucleotide Additions/Deletions patterns, amino acid distribution in the CDR3 region). This study provides chromosome-level annotations of IG loci in bats from different families and preliminarily reveals the fundamental characteristics of the bat BCR-CDR3 repertoire, along with key differences compared to human and mouse. Our findings offer comparative base-data, novel insights, tools, and strategies for future in-depth investigations into B-cell adaptive immune responses in bats.

BackgroundRhinolophus pusillus is currently the only known horseshoe bat that simultaneously harbors SARS-CoV-related coronaviruses (SARSr-CoVs), SARS-CoV-2-related coronaviruses (SC2r-CoVs), and a sarbecovirus recombinant lineage with a mosaic genome combining features of both SARS-CoV and SARS-CoV-2. However, the molecular basis underlying multi-virus coexistence and viral tolerance in this species remains incompletely defined. Here, we focus on basal gene expression patterns to investigate the immune landscape of this species.MethodsMulti-tissue transcriptomes were generated from six individuals of R. pusillus and Rhinolophus ferrumequinum, and integrated with publicly available RNA-seq datasets from multiple mammalian species relevant to zoonotic disease. Cross-species comparative transcriptomic analyses were performed to assess basal immune activity. Weighted gene co-expression network analysis (WGCNA) was applied to identify tissue-specific regulatory modules, and differential expression analyses were conducted to characterize immune-related transcriptional differences. In addition, long non-coding RNA (lncRNA)-mRNA regulatory networks were constructed to explore coordinated immune regulation.ResultsR. pusillus exhibited elevated basal transcriptional activity of innate antiviral immune processes compared with other mammals, with significant enrichment of genes involved in innate immune responses and antiviral defense. In contrast, transcriptional programs associated with inflammatory regulation were relatively attenuated. WGCNA identified distinct tissue-specific functional modules with coordinated regulatory architectures. R. pusillus displayed transcriptional features consistent with an enhanced capacity for viral infection tolerance relative to R. ferrumequinum. LncRNA analyses indicated coordinated regulation of inflammatory responses and DNA damage repair through lncRNA-mRNA regulatory networks.ConclusionsBy integrating comprehensive transcriptomic analyses of protein-coding RNAs with the prediction of lncRNA, this study provides a systematic molecular framework based on constitutive gene expression for understanding the mechanisms of viral tolerance in R. pusillus under conditions of multi-virus coexistence. These findings advance our understanding of bat immunobiology and offer insights into infection-related immune adaptations in R. pusillus as a key viral reservoir species.

Bats (Chiroptera) are the only mammals capable of true flight and rely heavily on echolocation for navigation, foraging, and predator avoidance in nocturnal environments. However, the mechanisms of echolocation signal regulation, its coevolution with morphology and ecology, and adaptive strategies under acoustic interference remain to be systematically integrated. This paper uses a literature review method to synthesize findings from 14 studies, exploring three core aspects: the structural and functional characteristics of the echolocation system, ecological and evolutionary drivers of signal diversity, and adaptive regulatory strategies under acoustic interference. The paper concludes that bat echolocation systems exhibit high plasticityConstant Frequency-Frequency Modulation bats (e.g., Rhinolophus ferrumequinum) adjust call components via Doppler shift compensation, while FM bats (e.g., Eptesicus fuscus) modify frequency and amplitude to avoid interference; echolocation signals coevolve with body size (forearm length) and jaw morphology, reflecting foraging niche differentiation; and anthropogenic noise and conspecific calls drive species-specific regulatory behaviors, with rural bats more sensitive to high-frequency insect noise and urban bats dominated by low-frequency anthropogenic noise. These findings provide a comprehensive framework for understanding bat sensory adaptation and inform conservation strategies for noise-affected bat populations.

Life-history trade-offs between reproduction and survival are well documented, yet the biological mechanisms underlying costs remain unclear. Telomere length (TL) is a potential biomarker for such costs, although its association with reproductive efforts is mixed. Bats, particularly the long-lived greater horseshoe bat (Rhinolophus ferrumequinum), provide a rare opportunity to explore these dynamics due to their longevity and low reproductive rates. We examined telomere dynamics in 202 female R. ferrumequinum (819 samples) from a wild population with over 65 y of monitoring, to assess whether reproductive effort leads to telomere shortening (cost of reproduction) or whether longer telomeres were associated with greater reproductive success (reflecting individual quality). Using Bayesian models, we show that females breeding from 2 y of age had significantly shorter relative TL (rTL) compared to females delaying reproduction until later ages. Selective disappearance was evident, with individuals possessing shorter rTL less likely to persist in the population. Cumulative reproductive success showed a positive nonsignificant association with rTL, consistent with the idea that long-term costs may be mitigated by individual quality or selective disappearance of low-quality individuals. However, short-term reproductive costs were evident, particularly in older females that bred in the previous year. Female R. ferrumequinum rTL declined during annual summer reproductive periods, particularly through the energetically demanding lactation stage. Within individuals, shorter rTL was associated with a reduced probability of surviving to the following year. These findings highlight the interplay between reproductive investment, telomere dynamics, and survival, supporting aspects of both the cost of reproduction and individual quality hypotheses in long-lived, low-fecundity species such as bats.

Abstract Hedgerows are key semi‐natural elements in European agricultural landscapes supporting diverse wildlife, including bats, that depend on these linear elements for foraging and commuting. Despite replanting initiatives, many hedgerows remain degraded due to intensive management practices such as overtrimming. To develop effective and generalisable conservation guidelines, more research is needed to understand how both management practices and intrinsic hedgerow characteristics influence bat species with different foraging strategies. We conducted passive acoustic monitoring across 448 hedgerows to assess the influence of six characteristics on bat activity: trimming method, height, width, foliage density, connectivity and woody plant diversity. We examined responses for two species associated with hedgerow landscapes ( Barbastella barbastellus , Rhinolophus ferrumequinum ) and three echolocation guilds: short‐, mid‐ and long‐range echolocators (SRE, MRE, LRE), corresponding to clutter, edge and open‐space foragers, respectively. Pollarding consistently increased bat activity, especially for tree‐dwelling species, likely by maintaining mature trees with larger crowns and diversified microhabitats. Coppicing generally reduced activity for SRE, B. barbastellus and R. ferrumequinum , while increasing LRE activity. Taller, tree‐filled hedgerows generally supported higher bat activity, whereas width and foliage density had more limited effects. Finally, uninterrupted hedgerow segments enhanced SRE and MRE activity, highlighting the importance of local habitat continuity in creating effective corridors for bat movement. Woody plant diversity was also positively associated with SRE activity. These results were mostly consistent across simplified open croplands and structurally complex bocage landscapes. Synthesis and applications . Our findings underscore the value of extensive hedgerow management in promoting on‐farm bat activity and support the integration of simple hedgerow quality assessment protocols into agri‐environment schemes. Consistent with previous research, we confirm the importance of tall, tree‐filled hedgerows and emphasise the need to preserve mature trees. We also provide novel insights: hedgerow continuity and low‐intensity trimming methods can further improve habitat suitability, particularly for tree‐dwelling bats. Our management recommendations align with evidence from other taxonomic groups, including birds and invertebrates, and can contribute to broader biodiversity conservation strategies.

Pseudogymnoascus destructans (Pd) invades the skin tissue of bats, leading to severe population declines. The skin microbiome plays a crucial role in protecting hosts from fungal infection and exhibits pronounced spatiotemporal dynamics in its structure and function. Meanwhile, metabolites derived from microbial communities reflect the host physiological state and participate in microbe-pathogen interactions. In this study, we investigated the spatiotemporal dynamics of skin bacterial communities and metabolites during hibernation in Rhinolophus ferrumequinum by integrating 16S rRNA sequencing with untargeted metabolomics and experimentally verified the antifungal effects of microbially derived potential metabolites against Pd. Our results revealed that the structure of the skin bacterial community varied significantly across sampling contexts, with its assembly primarily governed by stochastic processes. Bacterial diversity reached its lowest level during middle hibernation, accompanied by a simplified co-occurrence network dominated by cooperative or mutualistic interactions. Additionally, metabolomic analyses demonstrated systematic metabolic remodeling of bat skin across hibernation stages, marked by significant enrichment of multiple pathways closely involved in host antimicrobial defense. Furthermore, metabolite profiles differed across locations, and the abundance patterns of several metabolites were strongly correlated with Pd infection levels. Integrated analyses identified multiple metabolites that showed significant correlations with bacterial genera capable of synthesizing the corresponding compounds. In vitro validation confirmed that nine metabolites effectively inhibited the growth of Pd, among which melatonin exhibited the strongest antifungal activity. Collectively, this study reveals the dynamics of the skin microbiome and metabolites of R. ferrumequinum during hibernation, providing novel insights into the defensive role of skin-associated microbes and metabolites in maintaining population health and resilience against fungal pathogens.

The skull of amniotes is categorized into three conditions based on skeletal arrangement in the temporal region: anapsid, synapsid and diapsid. Mammals (class Mammalia), a descendent lineage of the clade Synapsida, possess the synapsid skull, which is characterized by a single lower temporal arch that ventrally borders the lower temporal fenestra. Although we previously suggested, based on the data from placental mammals, that the reduction in the expression domain of the upstream osteogenic genes Msx2 and Runx2 in the embryonic temporal mesenchyme might have played a role in the evolution of synapsid skulls, the molecular basis of synapsid skull evolution is still largely unknown. In this study, we investigated expression patterns of four osteogenic genes (two upstream genes Msx2 and Runx2 and two downstream genes Sp7 and Sparc) in the embryonic and neonatal temporal region of the gray short-tailed opossum Monodelphis domestica, the most commonly used experimental marsupial model, in order to more thoroughly understand the molecular basis of development of synapsid skulls unique to mammals. We found that M. domestica embryos and neonates display very restricted expressions of Msx2 and/or Runx2 in the dermal bone precursors in the temporal region, as two placental species do (the house mouse Mus musculus and the greater horseshoe bat Rhinolophus ferrumequinum). Spatially restricted expression of Msx2 and Runx2 in the embryonic temporal region may be a foundation for creating the "advanced" synapsid skull shared by all mammals where only three dermal bones configure the temporal region.

Bats are suspected reservoirs for several emerging viruses, including coronaviruses. Here, we report the detection of coronaviruses in the insectivorous bat, Rhinolophus ferrumequinum, in Ehime Prefecture, located in Shikoku, the smallest of the four main islands of Japan. Phylogenetic analysis of the partial sequence of the RNA-dependent RNA polymerase gene revealed that alphacoronaviruses have circulated throughout Ehime. Additionally, the near whole-genome sequence of the identified virus (EhRf2310-18) was closely related to that of BtRf-alphaCoV/HuB2013 identified in R. ferrumequinum in China (93.062% overall nucleotide identity), indicating that bat coronaviruses circulating in Ehime belong to the Decacovirus subgenus of Alphacoronavirus. Alphacoronaviruses in R. ferrumequinum may be strongly adapted to this species, and are distributed across a wide area between Ehime and China.

Demographic parameters such as survival and dispersal play a significant role in shaping population dynamics and genetic structure. However, dispersal remains difficult to estimate in natural populations, as it requires extensive individual sampling and monitoring in the landscape. We conducted a five-year capture-mark-recapture (CMR) study of Rhinolophus ferrumequinum in Nouvelle Aquitaine, France, monitoring 6673 individuals across 302 summer and winter roosts. For the first time in this species, we estimated seasonal survival and dispersal for both sexes using CMR models. We found high average adult survivals, with males displaying lower survival than females. Interestingly, both sexes showed high fidelity to summer or winter roosts with no strong marked differences between sexes. Strikingly, we found cues that females travelled on average longer distances than males to reach hibernation sites, challenging previous findings that dispersal is strongly male-biased in this species. Despite limited dispersal of both males and females, previous genetic studies have reported weak spatial genetic structure, suggesting that mating outside maternity colonies maintains gene flow. Our study highlights the importance of studying the full annual cycle monitoring in bats and supports conservation strategies that protect both maternity and hibernation roosts, as well as the landscape connectivity that enables seasonal movements.

Genetic diversity of pathogenic Leptospira spp. harbored by bats in Japan.

Coronaviruses, including SARS-CoV-2, can cause severe disease in humans, whereas reservoir hosts like Rhinolophus bats remain asymptomatic. To investigate how host-specific protein-protein interactions (PPIs) influence infection, we generated comparative PPI maps for SARS-CoV-2 and its bat-origin relative RaTG13 using affinity purification-mass spectrometry (AP-MS) in human and Rhinolophus ferrumequinum (RFe) bat cells. This approach identified both conserved and virus- and host-specific interactions that regulate infection dynamics. Notably, SARS-CoV-2 required a non-synonymous mutation in nucleocapsid to replicate in bat cells expressing human ACE2 and TMPRSS2. Analysis of the viral protein Orf9b revealed differential interactions with mitochondrial proteins Tom70 and MTARC2. A single residue difference in Orf9b between SARS-CoV-2 and RaTG13 functions as a molecular switch, weakening Tom70 binding and immune evasion in human cells while enhancing interaction with the bat-specific restriction factor MTARC2. These findings demonstrate how a single-residue substitution can reshape virus-host interactions and contribute to immune evasion and host adaptation.

ABSTRACTWhite‐nose syndrome (WNS), caused by the fungus Pseudogymnoascus destructans (Pd), has led to significant mortality and species endangerment in North America. Bats in eastern China, however, carry low loads of Pd and do not exhibit disease, suggesting natural resistance. To explore potential defenses, we isolated and screened antagonistic bacteria from the wing membranes of Rhinolophus ferrumequinum for their ability to inhibit Pd growth. We employed the plate delineation isolation method to obtain 74 single strains, which were then screened for antagonistic effects through contact and non‐contact inhibition experiments. A total of 18 antagonistic strains were isolated. After sequencing and comparison with the NCBI database, we identified eight known species and three unidentified species of antagonistic bacteria: Pseudomonas carnis, Buttiauxella ferragutiae, Paraburkholderia fungorum, Arthrobacter rhombi, Serratia liquefaciens, Paeniglutamicibacter gangotriensis, Brevibacterium aurantiacum, Acinetobacter lactucae, Pseudomonas sp., Brevibacterium sp., and Acinetobacter sp. Seventeen isolated strains showed varying degrees of inhibition by contact, while five species, including P. carnis, B. ferragutiae, S. liquefaciens, P. gangotriensis, and Brevibacterium sp., also inhibited Pd via non‐contact mode. We utilized solid‐phase microextraction coupled with GC–MS to obtain approximately 20 volatile compounds, all of which exhibited inhibitory effects on the growth of Pd, including ketones, aldehydes, and sulfur ethers. Notably, 5 ppm of 1‐undecene, dimethyl trisulphide, and 2‐nonanone each independently inhibited Pd growth. These findings suggest that the antagonistic strains and their VOCs might help protect bats from WNS. Understanding the interactions between Pd and skin flora, along with their VOCs, may be crucial in mitigating bats' vulnerability to WNS and developing effective mitigation strategies in the future.

Bats are the natural reservoirs for a variety of emerging and re-emerging viruses. Among them, rabies virus (genus Lyssavirus, family Rhabdoviridae) is one of the first and most emblematic described in these animals. Since its first description, several new bat lyssaviruses have been regularly identified. In addition to lyssaviruses, other bat rhabdoviruses have also been discovered, including members of the genera Vesiculovirus, Ledantevirus and, more recently, Alphanemrhavirus and Tupavirus. However, the family Rhabdoviridae is one of the most abundant and diverse viral families, with 434 officially recognized species, divided into 5 subfamilies and 56 different genera. The number of rhabdoviruses associated with bats is therefore probably higher than that currently available. In this study, we first developed and validated a combined nested RT-qPCR technique (pan-rhabdo RT-nqPCR) dedicated to the broad detection of animal rhabdoviruses. After validation, this technique was used for a large retrospective screening of archival bat samples (n = 1962), including blood (n = 816), brain (n = 723) and oral swab (n = 423). These samples were collected from various bat species over a 12-year period (2007-2019) in 9 different countries in Europe and Africa. A total of 23 samples (1.2%) from bat species Miniopterus schreibersii, Rhinolophus euryale and Rhinolophus ferrumequinum tested positive for rhabdovirus infection, including 17 (2.1%) blood and 6 (1.4%) oral swab samples, all collected from bats originating from the Mediterranean region. Complete virus genome sequences were obtained by next-generation sequencing for most of the positive samples. Molecular and phylogenetic analysis of these sequences demonstrated that the virus isolates, named Mediterranean bat virus (MBV), were closely related and represented a new species, Mediterranean vesiculovirus, within the genus Vesiculovirus. MBV was more specifically related to other bat vesiculoviruses previously described from China and North America, together clustering into a distinct group of bat viruses within this genus. Interestingly, our results suggest that MBV is widespread, at least in the western part of the Mediterranean region, where it circulates in the blood of several bat species. These results expand the host range and viral diversity of bat vesiculoviruses, and pave the way for further studies to determine the transmission route and dissemination dynamics of these viruses in bat colonies, as well as to assess their potential threat to public health.

Bats are the natural reservoirs for a variety of emerging and re-emerging viruses. Among them, rabies virus (genus Lyssavirus, family Rhabdoviridae) is one of the first and most emblematic described in these animals. Since its first description, several new bat lyssaviruses have been regularly identified. In addition to lyssaviruses, other bat rhabdoviruses have also been discovered, including members of the genera Vesiculovirus, Ledantevirus and, more recently, Alphanemrhavirus and Tupavirus. However, the family Rhabdoviridae is one of the most abundant and diverse viral families, with 434 officially recognized species, divided into 5 subfamilies and 56 different genera. The number of rhabdoviruses associated with bats is therefore probably higher than that currently available. In this study, we first developed and validated a combined nested RT-qPCR technique (pan-rhabdo RT-nqPCR) dedicated to the broad detection of animal rhabdoviruses. After validation, this technique was used for a large retrospective screening of archival bat samples (n = 1962), including blood (n = 816), brain (n = 723) and oral swab (n = 423). These samples were collected from various bat species over a 12-year period (2007–2019) in 9 different countries in Europe and Africa. A total of 23 samples (1.2%) from bat species Miniopterus schreibersii, Rhinolophus euryale and Rhinolophus ferrumequinum tested positive for rhabdovirus infection, including 17 (2.1%) blood and 6 (1.4%) oral swab samples, all collected from bats originating from the Mediterranean region. Complete virus genome sequences were obtained by next-generation sequencing for most of the positive samples. Molecular and phylogenetic analysis of these sequences demonstrated that the virus isolates, named Mediterranean bat virus (MBV), were closely related and represented a new species, Mediterranean vesiculovirus, within the genus Vesiculovirus. MBV was more specifically related to other bat vesiculoviruses previously described from China and North America, together clustering into a distinct group of bat viruses within this genus. Interestingly, our results suggest that MBV is widespread, at least in the western part of the Mediterranean region, where it circulates in the blood of several bat species. These results expand the host range and viral diversity of bat vesiculoviruses, and pave the way for further studies to determine the transmission route and dissemination dynamics of these viruses in bat colonies, as well as to assess their potential threat to public health.