Dead Bats Research Articles

Monitoring of Astroviruses, Brno-Hantaviruses, Coronaviruses, Influenza Viruses, Bornaviruses, Morbilliviruses, Lyssaviruses and Pestiviruses in Austrian Bats.

Here, we report the results of a monitoring study of bat viruses in Austria to strengthen the knowledge of circulating viruses in Austrian bat populations. In this study, we analyzed 618 oropharyngeal and rectal swab samples from 309 bats and 155 pooled tissue samples from dead bats. Samples were collected from 18 different bat species from multiple locations in Austria, from November 2015 to April 2018, and examined for astroviruses, bornaviruses, coronaviruses, hantaviruses, morbilliviruses, orthomyxoviruses (influenza A/C/D viruses), pestiviruses and rhabdoviruses (lyssaviruses) using molecular techniques and sequencing. Using RT-qPCR, 36 samples revealed positive or suspicious results for astroviruses, Brno-hantaviruses, and coronaviruses in nine different bat species. Further sequencing revealed correspondent sequences in five samples. In contrast, none of the tested samples was positive for influenza viruses A/C/D, bornaviruses, morbilliviruses, lyssaviruses, or pestiviruses.

Ectoparasites collected from dead bats (Chiroptera: Vespertilionidae) in east-central Poland.

During routine inspections of 4 bat shelters in central-eastern Poland, 36 dead bats belonging to 6 species were found: Myotis daubentonii, M. nattereri, M. myotis, Nyctalus noctula, Plecotus auritus and Eptescius serotinus. In the laboratory, 298 arthropods from 15 taxa (57 insects and 241 mites) were collected from these bats, and their relationships with their hosts were analysed. The first probable record of I. ricinus feeding on P. auritus is published.

Molecular detection of Babesia vesperuginis in bats from Lithuania

Babesia vesperuginis is an intraerythrocytic protozoan parasite that circulates among bats and ticks in many countries worldwide. However, the distribution of B. vesperuginis in the Baltic region has not been studied. A total of 86 dead bats from eight different species were collected and screened for Babesia spp. using real-time PCR. Overall, 52.3% (45/86) of the bats were found positive for Babesia spp. The prevalence of Babesia spp. in different organs varied, with the highest prevalence observed in heart tissues (37.0%) and the lowest in liver tissues (22.2%). However, the observed differences in prevalence among organs were not statistically significant. Blood samples from 125 bats of nine different species were also analyzed for Babesia spp. prevalence using real-time PCR and nested PCR. The results showed a prevalence of 35.2% and 22.4%, respectively. Moreover, 28.3% (17/60) of the examined blood samples were confirmed positive for Babesia spp. through blood smear analysis. The total of 32 partial sequences of the 18S rRNA gene derived in this study were 100% identical to B. vesperuginis sequences from GenBank. In eight species of bats, Pipistrellus nathusii, Pipistrellus pipistrellus, Pipistrellus pygmaeus, Vespertilio murinus, Eptesicus nilssonii, Eptesicus serotinus, Myotis daubentonii and Nyctalus noctula, Babesia parasites were identified. In E. nilssonii, Babesia spp. was identified for the first time.

Changes in diversity and species composition in the assemblage of live and dead bats at wind farms in a highly diverse region.

Besides direct mortality, wind farms also affect aerial fauna by modifying their communities, reducing species diversity and richness through disturbance. During three consecutive years, we used mist nets and acoustic recorders, and conducted carcass searches, to characterize the assemblage of bat species and to estimate bat mortality at two nearby wind farms sited <5 km apart in a highly biodiverse region. We asked whether the diversity, richness and evenness of the assemblages varied yearly, predicting it would decrease through time. Richness and evenness did not change, but the diversity of species recorded acoustically, 96% being aerial insectivores, was significantly lower the third year. We estimate 4 - 15.7 fatalities/MW/year by wind farm, with 63% of species found as carcasses being aerial insectivores. We found >40% of dissimilarity in the species composition of bat assemblages between wind farms despite the short distance between them, with species turnover accounting for more than half of the dissimilarity every year. Similarly, species turnover accounted for >15% of the dissimilarity in the composition of the assemblage of live bats (captured and recorded acoustically) and the assemblage obtained through carcass searches. Our findings suggest that nearby wind farms impact bat communities differentially and aerial insectivores disproportionally. Long term, multi-method surveys are needed to characterize bat communities in highly diverse regions and to evaluate the post-construction effects that wind farms have on them.

Cicada feast and food links: Grey-headed flying fox preys on black prince in south-East Australia

Cicada mass emergence offers a plentiful and energy-rich food source for a variety of predators, mainly birds and spiders. Megabats (Pteropodidae) mostly feed on nectar and fruits, but occasionally prey on insects such as sap-sucking cicadas and fruit-eating beetles. We observed and documented with photographs the behavioural repertoire of the Grey-headed Flying Fox Pteropus poliocephalus while hunting and consuming the Black Prince cicada Psaltoda plaga in Sydney, New South Wales, Australia. The bats used three main tactics while hunting the cicadas: a) flushing and/or catching cicadas on tree foliage while clambering over branches or landing on the foliage, b) lunging from a perch after spotting a resting cicada, and c) pursuing cicadas on the wing. Upon securing a cicada the bats flew off and landed on a feeding perch, where they chewed the prey and spat the remains as a pellet below this perch. On the ground, the pellets were usually sought by large ants that tore off pieces from these remains, the ants transporting them to the nest. Small ants aggregated on the pellets probably to seek the remaining sap, after which they left the pellets. The above described events showed three main food links: a) cicadas feeding on plant sap, b) bats preying on cicadas and spitting the remains as pellets, and c) ants feeding on bat pellets. The links illustrate a food chain that include plants, cicadas, bats, and ants. Additional food links we observed were: 1) cicadas feeding on different plants, 2) cicadas preyed on by diurnal animals, 3) dead cicadas consumed by ants, 4) dead bats consumed by birds, and 5) nocturnal raptorial bird that may prey on bats. This report seems the first to mention food links within a megabat-insect trophic interaction.

Discovery of a novel bat lyssavirus in a Long-fingered bat (Myotis capaccinii) from Slovenia.

Lyssaviruses are the causative agents of rabies, a zoonotic, fatal disease that is thought to be ancestral to bats. In the last decade, the detection of bat associated lyssaviruses is increasing also in Europe. Within a retrospective bat associated lyssavirus surveillance study a total of 225 dead bats of 21 bat species were collected in Slovenia between 2012 and 2019 and tested by specific real-time RT-PCR method. The first lyssavirus positive sample in bats in Slovenia was detected using the real-time RT-PCR, the fluorescent antibody test, and next generation sequencing, while the rabies tissue culture inoculation test was unsuccessful due to sample degradation and storage conditions. The nearly complete genome of Divača bat lyssavirus from Slovenia consists of 11,871 nucleotides and reflects the characteristic gene organization known for lyssaviruses, encoding the five viral proteins. Phylogenetic analysis of Divača bat lyssavirus revealed that it belongs to phylogroup I lyssaviruses and is most closely related to Kotalahti bat lyssavirus (KBLV) with 87.20% nucleotide and 99.22% amino acid identity. Together with KBLV, Khujand virus, European bat lyssavirus 2, Bakeloh bat lyssavirus, and Aravan virus, Divača bat lyssavirus was detected in the genus Myotis suggesting its key role in the transmission and maintenance of certain lyssaviruses.

Novel Bat Lyssaviruses Identified by Nationwide Passive Surveillance in Taiwan, 2018-2021.

Bat lyssaviruses were identified in Taiwan’s bat population during 2016–2017. The lyssavirus surveillance system was continuously conducted to understand the epidemiology. Through this system, the found dead bats were collected for lyssavirus detection by direct fluorescent antibody test and reverse transcription polymerase chain reaction. Three bats were identified as positive during 2018–2021. A novel lyssavirus, designated as Taiwan bat lyssavirus 2, was detected in a Nyctalus plancyi velutinus. This lyssavirus had less than 80% nucleotide identity in the nucleoprotein (N) gene with other lyssavirus species, forming a separate branch in the phylogenetic analysis. The other two cases were identified in Pipistrellus abramus (Japanese pipistrelles); they were identified to be similar to the former lyssavirus identified in 2016–2017, which was renominated as Taiwan bat lyssavirus 1 (TWBLV-1) in this study. Even though one of the TWBLV-1 isolates showed high genetic diversity in the N gene compared with other TWBLV-1 isolates, it may be a TWBLV-1 variant but not a new species based on its high amino acid identities in the nucleoprotein, same host species, and same geographic location as the other TWBLV-1.

Arachnids that feed on vertebrate carrion: necrophagy by the whip spider Paraphrynus raptator (Amblypygi: Phrynidae) and its relation to the feeding behavior of the woolly false vampire bat Chrotopterus auritus (Chiroptera: Phyllostomidae).

Necrophagy is a feeding strategy in which animals feed on carrion; most scavengers are facultative and can also be predators or consumers. For amblypygids, necrophagy is a poorly documented phenomenon and there are literature records of individuals of three different species feeding on dead bats inside caves. In the present note, we document for the first time a necrophagic behavior in the whip spider Paraphrynus raptator (Pocock, 1902) which was observed feeding on Otonyctomys hatti Anthony, 1932 (Rodentia: Cricetidae) and a yucatan poorwill, Nyctiphrynus yucatanicus Hartert, 1892 (Caprimulgiformes: Caprimulgidae) carrion. We made the observations inside a small chamber in an ancient Mayan temple inhabited by a group of woolly false vampire bats (Chrotopterus auritus Peters, 1856) in southeastern Mexico. Carrion consumption in P. raptator is directly related with the carnivorous feeding behavior of the C. auritus group with which they coexist.

The Use of Intrinsic Markers for Studying the Migratory Movements of Bats.

Simple SummaryMigratory bats species are among the most heavily impacted by the erection of wind energy facilities, with many individuals killed at wind turbines each year. Bat carcasses may be collected and used for a variety of biological studies. In this paper, we review the use of intrinsic markers—chemical signatures in bat tissues that can provide information about that animal’s life history—to study bat movements across the landscape. In doing so, we aim to provide our audience with a better understanding of the currently available literature and, more importantly, the areas of this field that need expansion. We emphasize the applications of intrinsic markers that have not been used extensively to study migratory bat species (i.e., trace elements, contaminants, strontium isotopes), and provide a workflow for researchers interested in conducting studies of this type.Mortality of migratory bat species at wind energy facilities is a well-documented phenomenon, and mitigation and management are partially constrained by the current limited knowledge of bat migratory movements. Analyses of biochemical signatures in bat tissues (“intrinsic markers”) can provide information about the migratory origins of individual bats. Many tissue samples for intrinsic marker analysis may be collected from living and dead bats, including carcasses collected at wind energy facilities. In this paper, we review the full suite of available intrinsic marker analysis techniques that may be used to study bat migration, with the goal of summarizing the current literature and highlighting knowledge gaps and opportunities. We discuss applications of the stable isotopes of hydrogen, oxygen, nitrogen, carbon, sulfur; radiogenic strontium isotopes; trace elements and contaminants; and the combination of these markers with each other and with other extrinsic markers. We further discuss the tissue types that may be analyzed for each and provide a synthesis of the generalized workflow required to link bats to origins using intrinsic markers. While stable hydrogen isotope techniques have clearly been the leading approach to infer migratory bat movement patterns across the landscape, here we emphasize a variety of lesser used intrinsic markers (i.e., strontium, trace elements, contaminants) that may address new study areas or answer novel research questions.

Thomas H. Kunz

Thomas H. Kunz

Israeli Rousettus aegyptiacus Pox Virus (IsrRAPXV) Infection in Juvenile Egyptian Fruit Bat (Rousettus aegyptiacus): Clinical Findings and Molecular Detection.

During 2019, five carcasses of juvenile Egyptian fruit bats (Rousettus aegyptiacus) were submitted to the Kimron Veterinary Institute. These bats exhibited typical poxvirus like lesion plaques of different sizes on the skin, abdomen and the ventral side of the wings. Clinical and histopathological findings suggested a poxvirus infection. Infectious virus was isolated from skin swabs, skin tissue and tongue of the dead bats and was further confirmed to be a Poxvirus by molecular diagnosis using PCR with pan-chordopoxviruses primers. All the dead bats were found positive for two Poxvirus genes encoding a metalloproteinase and DNA dependent DNA polymerase. In this study, a novel real time quantitative PCR (qPCR) assay was established to further confirmed the presence of specific poxvirus viral DNA in all pathologically tested tissues. Moreover, according to sequence analysis, the virus was found to be highly similar to the recently discovered Israeli Rousettus aegyptiacus Pox Virus (IsrRAPXV).

Fungi from dead arthropods and bats of Gomantong Cave, northern Borneo, Sabah (Malaysia)

Borneo is a biodiversity and ecotourism hotspot, yet one of its least-studied ecosystems is their limestone caves. Not many studies have been conducted on the role fungi play in tropical cave ecosystems, and no fungal surveys have been conducted in the caves of Sabah, Malaysia. Here, we assess the mycofloral diversity on bat and arthropod cadavers in one of the most popular ecotourism destinations of northern Borneo, Gomantong caves. Opportunistic sampling of cadavers within the Semud Hitam chamber of Gomantong cave yielded nine dead arthropods and four dead bats. Twenty-four culturable fungi were isolated, of which 14 morphological taxonomic units (MTU) were observed. Twelve of the 14 MTUs underwent molecular characterization of the ITS gene region to confirm identification. All fungi were Ascomycetes except for one Basidiomycete isolate. Aspergillus spp. had the highest occurrence (45.8%), followed by Penicillium spp. (25.0%), and Fusarium sp. (12.5%). Ceratobasidium sp., Diaporthe sp., Pestalotiopsis sp., and Xylaria feejeensis were isolated once each. No more than one fungal taxon was isolated from each arthropod cadaver, and not all arthropods yielded culturable fungi. Bat cadavers yielded 14 out of 24 isolates (58.3%), with the highest occurrence of the fungi sampled from their skin. Our results corroborate that bats and arthropods play a role in fungal dispersion and introduction in the cave because their exteriors are likely to harbor fungi they are exposed to in the environment. We also conclude that cadavers are important substrates for fungal growth and proliferation, perpetuating the role of fungi as important decomposers in caves. This study provides a baseline of information of the mycobiome of Bornean caves for future bioprospecting and potential biotechnological applications.

Identification of a Novel Yersinia enterocolitica Strain from Bats in Association with a Bat Die-Off That Occurred in Georgia (Caucasus).

Yersinia entercolitica is a bacterial species within the genus Yersinia, mostly known as a human enteric pathogen, but also recognized as a zoonotic agent widespread in domestic pigs. Findings of this bacterium in wild animals are very limited. The current report presents results of the identification of cultures of Y. entercolitica from dead bats after a massive bat die-off in a cave in western Georgia. The growth of bacterial colonies morphologically suspected as Yersinia was observed from three intestine tissues of 11 bats belonging to the Miniopterus schreibersii species. These three isolates were identified as Y. enterocolitica based on the API29 assay. No growth of Brucella or Francisella bacteria was observed from tissues of dead bats. Full genomes (a size between 4.6–4.7 Mbp) of the Yersinia strains isolated from bats were analyzed. The phylogenetic sequence analyses of the genomes demonstrated that all strains were nearly identical and formed a distinct cluster with the closest similarity to the environmental isolate O:36/1A. The bat isolates represent low-pathogenicity Biotype 1A strains lacking the genes for the Ail, Yst-a, Ysa, and virulence plasmid pYV, while containing the genes for Inv, YstB, and MyfA. Further characterization of the novel strains cultured from bats can provide a clue for the determination of the pathogenic properties of those strains.

Discovery of novel fungal species and pathogens on bat carcasses in a cave in Yunnan Province, China

ABSTRACT Virulent infectious fungal diseases, in natural and managed landscapes, are increasing. Fungal diseases in humans, animals and plants have caused die-off and extinction events and have become a threat to food security. A caving expedition in Yunnan Province, China, revealed two bat carcasses covered in fungal mycelia. Eleven fungal isolates were obtained from these bat carcasses, and morphological observations and multigene phylogenetic analyses revealed they were Fusarium incarnatum, Mucor hiemalis and Trichoderma harzianum and four new species, Mortierella rhinolophicola, M. multispora, M. yunnanensis and Neocosmospora pallidimors. One of the more alarming findings is that a number of infections related to Neocosmospora, previously associated with human and animal mycotoxicoses, are reported to be increasing, and here we present a new species from this genus, isolated from dead bats. Due to the ecosystem services provided by bats, and the close relationship between bats and humans, future research should focus on the impacts and significance of N. pallidimors to human and animal health, examining its pathogenicity and secondary metabolites. Taxonomic descriptions, color images of the habitat, in situ samples, microstructures and cultures are presented. SEM photographs of microstructures and phylogenetic trees showing the placement of new and known species are also provided.

Detection of BBLV in Natterer’s bat in Poland

Rabies is a zoonosis caused by RNA viruses belonging to the genus Lyssavirus, which includes 16 species. In Europe, the majority of rabies cases in bats are caused by European bat 1 lyssavirus (EBLV-1) associated mainly with serotine bats (Eptesicus serotinus and Eptesicus isabellinus). In 2010, Bokeloh bat lyssavirus (BBLV) was detected in Natterer’s bat (Myotis nattereri) in Germany and it was isolated several times in Germany and France in the following years. As required by the programme of passive rabies surveillance, the dead bat was sent to the laboratory and identified as Natterer’s bat on the basis of morphological criteria, which was confirmed by genetic identification using the sequencing of the cytochrome b gene. Brain tissue and salivary glands were collected for examination. In the brain, the direct immunofluorescence test revealed lyssavirus antigen. The presence of the infectious virus in brain tissue was confirmed by a tissue culture inoculation test. High levels of viral RNA were found in the brain and salivary glands. In order to determine the virus species, a complete genome sequence was obtained by next-generation sequencing of RNAs. The analysis of the full virus genome sequence confirmed the occurrence of BBLV in Poland and revealed that it showed the highest similarity (99.3%) to German isolates from Bavaria.

Indiana bat roosting behavior differs between urban and rural landscapes

Urbanization may negatively affect forest obligate bat species. We compared the roosting behavior of federally endangered Indiana bats (Myotis sodalis) in a fragmented site, located on the leading edge of a developing urban area (Indianapolis, IN), with a site of relatively contiguous forest cover in Indiana, USA, during the summers of 2013–2015. Bats in both sites used large standing dead trees and bat boxes with low levels of overhead canopy closure (fragmented site: 26.1 ± 11.0%, contiguous site: 20.1 ± 7.3%) as primary roosts (large groups of bats resident over many days). Between the 2 sites, secondary roosts (individual bats or small groups resident for only a few days) differed in canopy closure at the roost-scale (fragmented site: 66.9 ± 5.1%, contiguous site: 37.1 ± 9.3%); secondary roosts in the contiguous site resembled solar-exposed primary roosts. Bats in the fragmented site switched roosts less often (1.7 ± 0.5 switching events/bat, 5.8 ± 1.7 days of use/roosts) and traveled shorter distances between roosts (702 ± 211 m) than bats in the contiguous site (2.3 ± 0.6 switching events/bat, 2.4 ± 0.4 days of use/roosts; 1216 ± 337 m between roosts). This suggests that the fragmented site may have had low densities of available roosts, which may help explain the heavy use of bat boxes by bats in the fragmented site. These data suggest that urban populations of Indiana bats behave differently than those in rural areas, which managers should consider while managing for this species.

Bat (Mammalia: Chiroptera) road casualties in the Kresna Gorge, Bulgaria

The Kresna Gorge is a biodiversity hotspot with high conservation value not only for Bulgaria, but the Balkan Peninsula. In the gorge region, many rare, endemic, and protected species and habitats are found (Hubenov 2012). Since the beginning of the 21st century, several road and highway projects have threatened the ecological prosperity of this area. Intensive chiropterological research in the last 25 years (beginning in 1993) has revealed that the Kresna Gorge is an area of great importance for bats, providing habitats for more than 50% of the 33 species present in Bulgaria (Petrov 2001). There is growing evidence that roads negatively affect bats' populations (Altringham and Kerth 2016). Theoretical studies have shown that in some situations, collisions might lead to population decline (see references in Altringham and Kerth 2016). Identifying parts of the road where bats experience collisions more frequently can have strong conservation implications. Between 2013 and 2016 we recorded roadkilled bats on a weekly basis, from March to October. We monitored a 16 km section of a high-traffic intensity road passing through the Kresna Gorge. We recorded 78 dead bats on the road, belonging to 10 species. During our survey, we recorded Myotis brandtii (Eversmann, 1845), a new species for the bat fauna of the Kresna Gorge. The presence of two species, Nyctalus leisleri (Kuhl, 1817) and Miniopterus schreibersii (Kuhl, 1817), previously with single or doubtful records, were confirmed. Statistically significant hotspots of roadkill were modelled using Getis-Ord Gi statistic. We identified five road stretches as hotspots for bat accidents. They are all relatively close to bridges, tunnel exits, and/or bat roosts. Оur study adds new data on the bats' species composition in the gorge, as well as it demonstrates that road elements in this section of the Pan-European Corridor IV concentrate and increase road accidents with bats.

Standard Operating Procedure for Lyssavirus Surveillance of the Bat Population in Taiwan.

Viruses within the genus Lyssavirus are zoonotic pathogens, and at least seven lyssavirus species are associated with human cases. Because bats are natural reservoirs of most lyssaviruses, a lyssavirus surveillance program of bats has been conducted in Taiwan since 2008 to understand the ecology of these viruses in bats. In this program, non-governmental bat conservation organizations and local animal disease control centers cooperated to collect dead bats or bats dying of weakness or illness. Brain tissues of bats were obtained through necropsy and subjected to direct fluorescent antibody test (FAT) and reverse transcription polymerase chain reaction (RT-PCR) for detection of lyssavirus antigens and nucleic acids. For the FAT, at least two different rabies diagnosis conjugates are recommended. For the RT-PCR, two sets of primers (JW12/N165-146, N113F/N304R) are used to amplify a partial sequence of the lyssavirus nucleoprotein gene. This surveillance program monitors lyssaviruses and other zoonotic agents in bats. Taiwan bat lyssavirus is found in two cases of the Japanese pipistrelle (Pipistrellus abramus) in 2016-2017. These findings should inform the public, health professionals, and scientists of the potential risks of contacting bats and other wildlife.

Filiviruses of southeast Asia, China and Europe (review)

Filoviruses are known as causative agents of severe haemorrhagic fevers with a high mortality rate in humans. Zaire ebolavirus and Marburgvirus, the most known of them, are associated with the occurrence of sporadic cases and outbreaks of hemorrhagic fevers in some parts of Africa. Isolation of Reston ebolavirus in 1989 in the United States from samples of dead cynomolgus monkeys imported from the Philippines was the first evidence on the existence of filoviruses outside the Africa. Due to the development of new diagnostic methods, Reston ebolavirus or its markers (RNA, antibodies) were found in different animals in the Philippines, China and some other countries of Southeast Asia. These events significantly changed the concept of the geography of filoviruses at present time. Novel filoviruses have been identified in bats in China using of molecular genetic methods. Detection of filovirus RNA (the Lloviu virus) in samples from dead common bent-winged bats in Spain (2002) and in Hungary (2016) indicates the possibility of circulation of filoviruses with unknown pathogenicity potential for humans and animals among bats of temperate latitudes. This review summarizes data on findings of filovirus markers in animals in Southeast Asia, China and Europe.

Assessment of a Rabies Virus Rapid Diagnostic Test for the Detection of Australian Bat Lyssavirus.

Australian bat lyssavirus (ABLV) is closely related to the classical rabies virus and has been associated with three human fatalities and two equine fatalities in Australia. ABLV infection in humans causes encephalomyelitis, resulting in fatal disease, but has no effective therapy. The virus is maintained in enzootic circulation within fruit bats (Pteropid spp.) and at least one insectivorous bat variety (Saccolaimus flaviventris). Most frequently, laboratory testing is conducted on pteropodid bat brains, either following a potential human exposure through bites, scratches and other direct contacts with bats, or as opportunistic assessment of sick or dead bats. The level of medical intervention and post-exposure prophylaxis is largely determined on laboratory testing for antigen/virus as the demonstrable infection status of the in-contact bat. This study evaluates the comparative diagnostic performance of a lateral flow test, Anigen Rabies Ag detection rapid test (RDT), in pteropodid variant of ABLV-infected bat brain tissues. The RDT demonstrated 100% agreement with the reference standard fluorescent antibody test on 43 clinical samples suggesting a potential application in rapid diagnosis of pteropodid variant of ABLV infection. A weighted Kappa value of 0.95 confirmed a high level of agreement between both tests.