Bat Species Research Articles

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

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

All-you-can-eat buffet: A spider-specialized bat species (Myotis emarginatus) turns into a pest fly eater around cattle.

Determining the dietary spectrum of European insectivorous bats over time is the cornerstone of their conservation, as it will aid our understanding of foraging behavior plasticity in response to plummeting insect populations. Despite the global decline in insects, a restricted number of arthropod pest species thrive. Yet past research has overlooked the potential of European bats to suppress pests harmful to woodlands or livestock, in spite of their economic relevance. Here we investigated the diet composition, its breeding season variations and pest consumption of an insectivorous bat species (Myotis emarginatus), at the northern edge of its range (Wallonia, Belgium). We also explored the prey ecology to gain insight into the hunting strategies and foraging habitats of this bat species. We used DNA metabarcoding to amplify two COI markers within 195 bat droppings collected in June, July and August, thereby identifying 512 prey taxa predominated by Diptera, Araneae and Lepidoptera. Overall, in 97% of the samples we detected at least one of the 58 potential pest taxa, 41 of which targeting trees. The June samples were marked by a diet rich in orb-weaver spiders, in accordance with the archetypal diet of M. emarginatus bats. However, during the highly energy demanding July-August parturition and lactation period, roughly 55% of the dropping samples contained two cattle fly pests (Stomoxys calcitrans and Musca domestica). Moreover, among the 88 Diptera species preyed upon by M. emarginatus in July and August, these flies accounted for around 50% of the taxa occurrences. This plasticity-the switch from a spider-rich to a fly-rich diet-seems providential considering the dramatic ongoing drop in insect populations but this involves ensuring bat-friendly cattle farming. Our results revealed that bats widely consume pest entomofauna, thereby highlighting their potential role as allies of forest managers and farmers.

Unexpected bat community changes along an urban–rural gradient in the Berlin–Brandenburg metropolitan area

Urbanization gradients are increasingly used in ecological studies to discover responses of species communities to different intensities of human-induced habitat transformation. Here, we investigated patterns of bat communities against the background of different urbanization levels using a priori defined urbanization categories based on distance classes (5 km intervals) along a linear transect from the urban core of the city of Berlin westwards into the rural outskirts of the state of Brandenburg. Using linear-mixed effects models, we found that “distance class”, as a proxy for urbanization level, is a meaningful and suitable predictor of bat species richness and diversity. We observed an unexpectedly sudden increase in bat species richness and diversity and changes in species-specific activity levels relatively close to the urban center at the transition between urban and peri-urban areas. This change suggests a relevant influence of the peri-urban areas as a “buffer zone” for specific bat species not able to adapt to the heavily modified inner core of the metropolitan area. Although we could demonstrate that anthropogenic noise and artificial light have the potential to predict the variability of bat species activity along the urban–rural gradient, the actual influence on observed shifts in the bat community needs further research.

Bats (Mammalia: Chiroptera) from two priority areas for biodiversity conservation in the Brazilian Amazon and range extension for Carollia benkeithi (Phyllostomidae)

Abstract Amazonian tropical forests host the highest bat diversity in South America. The central area Brazilian Amazon encompasses one of the largest pristine areas within this biome, where Amanã and Mamirauá Sustainable Development Reserves (SDRs) are located. These two reserves hold the status of Priority Areas for Biodiversity Conservation due to their high biological relevance. The aim of this study was to identify the bat species richness within these SDRs to minimize the Wallacean shortfall of the chiropterofauna in the Amazon. A total of 274 specimens, collected 20 years ago and housed in the mammal collection at the Museu Paraense Emílio Goeldi, were sorted and identified. They are distributed across four families, 18 genera, and 26 species: Artibeus planirostris and Carollia perspicillata were the most abundant vouchers. We identified a range extension for Carollia benkeithi and a possible new taxon for the genus Micronycteris.

Historical biogeography of the Neotropical noctilionoid bats (Chiroptera: Noctilionoidea), revisited through a geographically explicit analysis.

Biogeographic studies have generally relied on methods that use a few, large predefined areas, which may overlook fine-scale patterns. Here we test previous hypotheses about the biogeographic history of a diverse bat clade regarding its association with major Neotropical geological formations, particularly the Antilles, the South American Dry Diagonal, the Andes and the Panamanian land bridge, by applying a recently available method that uses actual distributions instead of predefined areas. We compiled and curated spatially explicit, georeferenced data of 173 bat species (Mammalia: Chiroptera: Noctilionoidea) from the online database Global Biodiversity Information Facility. By taking a previous comprehensive phylogeny as an evolutionary framework, we performed computationally intensive analyses using the Geographically-explicit Event Model. This method uses the observed species distributions to reconstruct the ancestral areas and biogeographic events at each phylogeny node. We found that sympatric speciation was the most frequently reconstructed event, and involved mainly the Panamanian Isthmus and northern South America (SA), but all sympatry reconstructions were different and specific to each node. Allopatric events were important in the Andes; vicariance caused both west/east and north/south disjunctions that went unnoticed previously. Founder events indicated bidirectional dispersal between the mainland and the Antilles since the Miocene, and across the incomplete Panamanian bridge and the SA Dry Diagonal since the early Pliocene. Overall, we found support for previous hypotheses on the influence of major Neotropical paleogeographic events in the diversification of the group, but additionally revealed multi-scale patterns that are embedded within the mainland and were previously overlooked. Our results highlight a trans-isthmian centre of diversification in the biogeographic history of Noctilionoidea including the Panamanian Isthmus and Northern SA.

Review and meta-analysis of correlates of home range size in bats

Abstract Home range is an ecological concept that affects many aspects of the life of vertebrates and hence understanding how it varies between species is crucial. Mammalian home range size has been linked to body size and diet, but these studies were based predominantly on terrestrial species and most specifically excluded bats. As the only group of flying mammals, bats experience distinctly different constraints on movement, and hence home range. However, despite their diversity, relatively little is known about the home ranges of bats, and this is the first global review assessing correlates of their home range size. Our hypothesis is that home range will be impacted by different ecological conditions experienced by the bats and by the biological traits of the bats themselves. We performed a meta-analysis based on published data for 81 bat species to identify variables that contribute most to home range size. Sex, wing loading, functional group, colony size, dietary class, distance from the equator (latitudinal region), habitat type, and the interaction between habitat type and latitudinal region were all important explanatory variables. Wing loading was positively correlated with home range size—while females, open-air foragers, large colony sizes, and bats in temperate regions consistently had large home ranges. Understanding the correlates of home range has important implications, for example, for bat conservation and for assessing the risk of spillover of zoonotic pathogens from bats to humans and livestock.

Interaction of sound-audition traits between eared insects and arthropodophagous bats: using a DNA approach to assess diet

Abstract Arthropod–bat interactions are often considered as a base model for studying factors underlying predator–prey coevolutionary processes. Bats developed ultrasonic echolocation to hunt, and in response some arthropods developed defense mechanisms such as ultrasonic hearing, allowing them to elude bat predators. The present study analyzes the feeding patterns of bats, focusing on sonic-auditory sensory mechanisms in predator–prey interactions. Next-generation DNA sequence data from fecal samples were used to analyze the diet of 17 bat species from Mexico. Arthropod prey taxa were classified according to their auditory traits, and echolocation data were recompiled from literature review. We: (i) classified arthropod families according to their hearing ability; (ii) estimated arthropod taxon richness and proportion in the diet of each bat species; and (iii) used multidimensional scaling, principal component analysis, and regression to analyze prey consumption patterns in relation to their auditory traits and in relation to echolocation characteristics of bats. Finally, we analyzed the relationship between foraging time and auditory characteristics of prey. Families with hearing organs correspond to the orders Lepidoptera and Orthoptera. We registered 20 families of Lepidoptera and 5 of Orthoptera—7 and 3 with hearing organs, respectively. Of these orders, families lacking ears were recorded in the diet of a few bat species. Our results support the allotonic frequency hypothesis predicting a difference in emission frequency intervals between predator and prey. However, we found that the consumption of earless moths is less frequent and is related to diurnal and twilight activity—hence, their consumption is limited to bat species foraging early. Results indicate bats feed on arthropod prey successfully despite the ultrasonic hearing ability of the prey. These results may be due to counteradaptations that allow maintenance of an asymmetric “arms race” between bats and eared insects that favors the predator.

Hibernacula of bats in Mexico, the southernmost records of hibernation in North America

Abstract Although Mexico holds the southernmost hibernating bats in North America, information on winter behavior and hibernacula microclimate use of temperate Mexican bats is limited. We studied hibernating bats at high altitudes (>1,000 m a.s.l.) in northern and central Mexico during 5 consecutive winters. Our aims were to document and describe the hibernacula, winter behavior (such as abundance and roost pattern), and microclimates (estimated as adjacent substrate temperature) of cave-hibernating bats in Mexico. We found 78 hibernacula and 6,089 torpid bats of 10 vespertilionid species, increasing by over 50% the number of cave-hibernating bat species and quadrupling the number of hibernacula for Mexico. Hibernacula were at altitudes between 1,049 and 3,633 m a.s.l., located in 3 mountain ranges, mainly in oak and conifer forests. Myotis velifer was the most common species, followed by Corynorhinus townsendii and C. mexicanus. We recorded the adjacent substrate temperatures from 9 species totaling 1,106 torpid bats and found differences in microclimate use among the 3 most common species. In general, abundance of torpid bats in our region of study was similar to those in the western United States, with aggregations of tens to a few hundred individuals per cave, and was lower than in the eastern United States where a cave may hold thousands of individuals. Knowledge of bat hibernation is crucial for developing conservation and management strategies on current conditions while accommodating environmental changes and other threats such as emerging diseases.

Preference for hibernacula microclimates varies among 3 bat species susceptible to white-nose syndrome

Abstract North American bat populations have experienced precipitous declines since the introduction of white-nose syndrome (WNS). Evidence that bats have responded to WNS by altering their winter habitat selection has been used to manipulate underground environments and improve winter survival. However, such management practices must be based on local data, which do not exist for all regions. Our goal was to identify determinants of winter habitat use for 3 bat species with varying susceptibility to WNS. To do so, we conducted 188 surveys of 129 potential hibernacula over 3 winters. We found that the range of winter temperatures and presence of water were important for the Little Brown Myotis (Myotis lucifugus) and Tricolored Bat (Perimyotis subflavus), species that are highly susceptible to WNS. Counts of both species were greater in sites with a narrow (<5 °C) temperature range and where streams or water bodies >25 m2 were present. In addition, we found larger groups of Little Brown Myotis in abandoned railroad tunnels than in caves. Winter counts of these species increased over time at 3 railroad tunnels that were surveyed for 3 consecutive winters, whereas populations were extirpated from historically important caves and mines. Counts of Big Brown Bats (Eptesicus fuscus), which are less susceptible to WNS, were also greater in hibernacula with water bodies. However, average winter temperature also helped predict counts of Big Brown Bats. This species had a curvilinear response to temperature, with most bats found at hibernacula with average winter temperatures of 4 to 6 °C, and fewer bats observed at warmer and colder conditions. These data show that focusing solely on historically important hibernacula may fail to achieve conservation goals for the Little Brown Myotis and Tricolored Bat in our region, and suggest that anthropogenic structures can provide habitat for remnant populations of imperiled bat species.

Presence of Trypanosoma cruzi TcI and Trypanosoma dionisii in sylvatic bats from Yucatan, Mexico.

Chagas disease is caused by Trypanosoma cruzi, whose genetic structure is divided into six discrete typing units (DTUs) known as TcI-TcVI. In the Yucatan Peninsula, Mexico, information regarding the DTUs circulating in wild mammals is scarce, while this is important knowledge for our understanding of T. cruzi transmission dynamics. In the current study, we sampled wild mammals in a sylvatic site of the Yucatan Peninsula and assessed their infection with T. cruzi by PCR. Then, for infected mammals, we amplified and sequenced nuclear and mitochondrial T. cruzi genetic markers for DTU identification. In total, we captured 99 mammals belonging to the orders Chiroptera, Rodentia and Didelphimorphia. The prevalence of infection with T. cruzi was 9% (9/99; 95% CI [5, 16]), and we identified TcI in a Jamaican fruit bat, Artibeus jamaicensis. Moreover, we fortuitously identified Trypanosoma dionisii in another Jamaican fruit bat and detected an unidentified Trypanosoma species in a third specimen. While the latter discoveries were not expected because we used primers designed for T. cruzi, this study is the first to report the identification of T. dionisii in a bat from Yucatan, Mexico, adding to a recent first report of T. dionisii in bats from Veracruz, and first report of this Trypanosoma species in Mexico. Further research is needed to enhance our knowledge of T. cruzi DTUs and Trypanosoma diversity circulating in wildlife in Southeastern Mexico.

Bat2Web: A Framework for Real-Time Classification of Bat Species Echolocation Signals Using Audio Sensor Data.

Bats play a pivotal role in maintaining ecological balance, and studying their behaviors offers vital insights into environmental health and aids in conservation efforts. Determining the presence of various bat species in an environment is essential for many bat studies. Specialized audio sensors can be used to record bat echolocation calls that can then be used to identify bat species. However, the complexity of bat calls presents a significant challenge, necessitating expert analysis and extensive time for accurate interpretation. Recent advances in neural networks can help identify bat species automatically from their echolocation calls. Such neural networks can be integrated into a complete end-to-end system that leverages recent internet of things (IoT) technologies with long-range, low-powered communication protocols to implement automated acoustical monitoring. This paper presents the design and implementation of such a system that uses a tiny neural network for interpreting sensor data derived from bat echolocation signals. A highly compact convolutional neural network (CNN) model was developed that demonstrated excellent performance in bat species identification, achieving an F1-score of 0.9578 and an accuracy rate of 97.5%. The neural network was deployed, and its performance was evaluated on various alternative edge devices, including the NVIDIA Jetson Nano and Google Coral.

Nation-wide surveillance of ticks (Acari: Argasidae) on bats (Chiroptera) in Singapore

Bats and ticks are important sources of zoonotic pathogens. Therefore, understanding the diversity, distribution, and ecology of both groups is crucial for public health preparedness. Soft ticks (Argasidae) are a major group of ectoparasites commonly associated with bats. The multi-host life cycle of many argasids make them important vectors of pathogens. Over nine years (2011–2020), surveillance was undertaken to identify the ticks associated with common bats in Singapore. During this period, the bat tick Ornithodoros batuensis was detected within populations of two cave roosting bat species: Eonycteris spelaea and Penthetor lucasi. We examined the relationship between bat species, roosting behaviour, and probability of O. batuensis infestation. We also estimated the relationship between bat life history variables (body condition index, sex, and age) on the probability of infestation and tick count. This represents the first detection of O. batuensis and the genus Ornithodoros within Singapore. We also provide evidence of the continued persistence of Argas pusillus in Singapore with the second local record.

Apparent absence of Trypanosoma cruzi in Mexican free-tailed bats (Tadarida brasiliensis) from Texas, USA

The Mexican free-tailed bat (Tadarida brasiliensis) is one of the most abundant mammals in North America. Mexican free-tailed bats have a wide geographic range stretching from northern South America to the western United States. Bats are theorized to be the original hosts for Trypanosoma cruzi —the causative agent of Chagas disease— and can serve as a source of infection to triatomine insect vectors that feed upon them. Chagas disease is a neglected tropical disease across the Americas where triatomines are present, including the southern United States, where Texas reports this highest number of locally-acquired human cases. To learn more about the role of bats in the ecology of Chagas disease in Texas, we surveyed a colony of Mexican free-tailed bats from Brazos County, Texas, for T. cruzi using carcasses salvaged after an extreme weather event. A total of 283 Mexican free-tailed bats collected in February 2021 were dissected and DNA from the hearts and kidneys was used for T. cruzi detection via qPCR. None of the bat hearts or kidneys tested positive for T. cruzi; this sample size affords 95% confidence that the true prevalence of T. cruzi in this population does not exceed 1%. Future sampling of multiple bat species as well as migrant and resident colonies of Mexican free-tailed bats across different times of the year over a broader geographic range would be useful in learning more about the role of bats in the ecology of Chagas disease in Texas.

Identification and characterization of recent retrovirus in Rhinolophus ferrumequinum bats.

An investigation into retrovirus was conducted in six species of bats (Myotis aurascens, Myotis petax, Myotis macrodactylus, Miniopterus fuliginosus, Rhinolophus ferrumequinum, and Pipistrellus abramus) inhabiting South Korea. Exogenous retroviruses (XRVs) were detected in the tissue samples of R. ferrumequinum individuals by PCR assay. Proviruses were identified in all tissue samples through viral quantification using a digital PCR assay per organ (lung, intestine, heart, brain, wing, kidney, and liver), with viral loads varying greatly between each organ. In phylogenetic analysis based on the whole genome, the Korean bat retroviruses and the R. ferrumequinum retrovirus (RfRV) strain formed a new clade distinct from the Gammaretrovirus clade. The phylogenetic results determined these viruses to be RfRV-like viruses. In the Simplot comparison, Korean RfRV-like viruses exhibited relatively strong fluctuated patterns in the latter part of the envelope gene area compared to other gene areas. Several point mutations within this region (6,878-7,774 bp) of these viruses were observed compared to the RfRV sequence. One Korean RfRV-like virus (named Y4b strain) was successfully recovered in the Raw 264.7 cell line, and virus particles replicated in the cells were confirmed by transmission electron microscopy. RfRVs (or RfRV-like viruses) have been spreading since their first discovery in 2012, and the Korean RfRV-like viruses were assumed to be XRVs that evolved from RfRV.IMPORTANCER. ferrumequinum retrovirus (RfRV)-like viruses were identified in greater horseshoe bats in South Korea. These RfRV-like viruses were considered exogenous retroviruses (XRVs) that emerged from RfRV. Varying amounts of provirus detected in different organs suggest ongoing viral activity, replication, and de novo integration in certain organs. Additionally, the successful recovery of the virus in the Raw 264.7 cell line provides strong evidence supporting their status as XRVs. These viruses have now been identified in South Korea and, more recently, in Kenya since RfRV was discovered in China in 2012, indicating that RfRVs (or RfRV-like viruses) have spread worldwide.

Identifikasi Ektoparasit pada Kelelawar di Gua Kenculit Desa Mekar Sari Kabupaten Lombok Tengah

Kenculit cave is one of the caves in the southern region of Lombok Island which is known as a habitat for several types of small mammals, one of which is bats (order Chiroptera). The problem is that bats are one of the mammals that act as hosts for ectoparasites. The aim of the research was to determine the types of ectoparasites in bats in the kenculit cave in Mekar Sari Village, West Praya District, Central Lombok Regency. This type of research is an exploratory descriptive study designed using direct observation. The bat sampling technique uses the trapping method. Meanwhile, ectoparasite samples were taken by combing the bat's hair and then observing using a stereo microscope. Based on the research results, it shows that there are 3 types of ectoparasites from 2 different bat species, namely Taphozous melanopagon with 1 type of ectoparasite (Amblyomma sp.), and the Miniopterus pusillus bat species found with 2 types of ectoparasite (Cyclopodia sp. and Ancystropus sp.).

Bat diversity in the Cuc Phuong National Park, Vietnam - Results from VIETBIO field training and annotated species list.

Biodiversity surveys are essential for both academic research and conservation. Integrative approaches that combine morphological, genetic and acoustic aspects for species identification can provide reliable information in taxonomy and evolution. This is especially relevant for those groups with a high degree of cryptic diversity such as bats. Here, we present the results from a field survey carried out in the Cuc Phuong National Park (CPNP) during 2019 as part of the VIETBIO project and from the examination of specimen collections preserved at the museums of CPNP and the Institute of Ecology and Biological Resources (IEBR). In addition, we include an annotated species list, based on this survey and a literature review. We here confirm that CPNP is home to at least 47 bat species belonging to 23 genera and seven families. We recorded ten of these bat species during our field survey. Obtained data in genetics (sequencing a fragment of the mitochondrial gene COI) supported the morphological identification of the recorded species for which we were able to produce these data. In addition, we include echolocation recordings obtained during our field training with the hope that they may contribute valuable insights to future work concerning the surveyed species. Results from the field survey represent a relevant contribution to biodiversity assessment efforts and, thus, support conservation and management efforts to maintain bat diversity in Vietnam.

Molecular detection of Bartonella spp. in bat-associated macronyssid mites (Acari: Macronyssidae) from Southern and Southeastern Brazil.

Despite the worldwide occurrence and high genetic diversity of Bartonella spp. in bats, few studies investigate their occurrence in bat-associated mites. To date, 26 species of Macronyssidae mite species have been reported from Brazil, and 15 of which were found parasitizing bats. The present study aimed to investigate the presence of Bartonella DNA in bat-associated macronyssid mites from Brazil. For this purpose, 393 macronyssid specimens were selected by convenience from the tissue bank of the Acari Collection of the Instituto Butantan (IBSP). These mites were collected from 14 different bat species in three different Brazilian States (Minas Gerais, Paraná, and Rio de Janeiro). Out of 165 mites positive in the PCR for the endogenous 18S rRNA gene, only eight were positive in the qPCR for Bartonella spp. based on the nuoG gene, and we were able to obtain two sequences base in this same gene, and one sequence based on the 16S rRNA gene. The phylogenetic inference based on the nuoG gene grouped the obtained sequences with Bartonella genotypes previously detected in bats and associated bat flies, while the phylogeny based on the 16S rRNA grouped the obtained sequence in the same clade of Bartonella genotypes previously detected in Dermanyssus gallinae. These findings suggest that macronyssid mites might be associated with the maintenance of bartonellae among bats.

Sosuga Virus Detected in Egyptian Rousette Bats (Rousettus aegyptiacus) in Sierra Leone.

Sosuga virus (SOSV), a rare human pathogenic paramyxovirus, was first discovered in 2012 when a person became ill after working in South Sudan and Uganda. During an ecological investigation, several species of bats were sampled and tested for SOSV RNA and only one species, the Egyptian rousette bat (ERBs; Rousettus aegyptiacus), tested positive. Since that time, multiple other species have been sampled and ERBs in Uganda have continued to be the only species of bat positive for SOSV infection. Subsequent studies of ERBs with SOSV demonstrated that ERBs are a competent host for SOSV and shed this infectious virus while exhibiting only minor infection-associated pathology. Following the 2014 Ebola outbreak in West Africa, surveillance efforts focused on discovering reservoirs for zoonotic pathogens resulted in the capture and testing of many bat species. Here, SOSV RNA was detected by qRT-PCR only in ERBs captured in the Moyamba District of Sierra Leone in the central region of the country. These findings represent a substantial range extension from East Africa to West Africa for SOSV, suggesting that this paramyxovirus may occur in ERB populations throughout its sub-Saharan African range.

A tale of endurance: bats, viruses and immune dynamics.

The emergence of highly zoonotic viral infections has propelled bat research forward. The viral outbreaks including Hendra virus, Nipah virus, Marburg virus, Ebola virus, Rabies virus, Middle East respiratory syndrome coronavirus, SARS-CoV and the latest SARS-CoV-2 have been epidemiologically linked to various bat species. Bats possess unique immunological characteristics that allow them to serve as a potential viral reservoir. Bats are also known to protect themselves against viruses and maintain their immunity. Therefore, there is a need for in-depth understanding into bat-virus biology to unravel the major factors contributing to the coexistence and spread of viruses.

Molecular prevalence of Bartonella spp. in bat flies in east coast Malaysia

Bats are a significant reservoir for numerous pathogens, including Bartonella spp. It is one of the emerging zoonotic bacterial diseases that can be transmitted to humans and may cause various unspecific clinical manifestations. Thus, bartonellosis is rarely diagnosed and is regarded as a neglected vector-borne disease (VBD). Bat flies have been hypothesised to be a vector in the transmission of pathogens among bats. They are host-specific, which reduces the likelihood of pathogen transmission across bat species; however, they are likely to maintain high pathogen loads within their host species. To explore the presence of Bartonella spp. in bat flies from Peninsular Malaysia; bat fly samples collected from various sites at the east coast states were subjected to molecular detection for Bartonella spp. It was discovered that 38.7 % of bats from Terengganu and Kelantan were infested with bat flies; however, no bat fly was found in bats collected from Pahang. The collected bat flies belonged to the families Nycteribiidae (79.6 %) and Streblidae (20.4 %). The collected bat flies were pooled according to the locations and species into 39 pools. Out of these 39 pools, 66.7 % (n = 26) were positive for Bartonella spp. by PCR. Sequence analyses of five randomly selected PCR-positive pools revealed that pools from Kelantan (n = 3) have the closest sequence identities (99 %) to Bartonella spp. strain Lisso-Nig-922 from Nigeria. However, the other pools from Terengganu (n = 2) were closely related to Bartonella spp. strain KP277 from Thailand and Bartonella spp. strain Rhin-3 from the Republic of Georgia with 99 % and 100 % sequence identity, respectively. This suggests that the Bartonella spp. found in Malaysian bat flies are genetically diverse and can potentially serve as reservoirs for pathogenic Bartonella spp.