Mating without intromission in a bat.

Copulatory behaviours stand as cornerstones of sexual selection, yet they remain mysterious in many species. Because of their nocturnal and elusive lifestyle, the copulatory behaviours of bats have been mostly overlooked1. Several aspects of bat reproduction differ from other mammals (e.g. prolonged sperm storage2, delayed development3). Here, we show that in serotine bats (Eptesicus serotinus) the penis is used as a 'copulatory arm' rather than an intromittent organ, revealing a novel copulatory behaviour in mammals.

Could a city-dwelling bat (Pipistrellus kuhlii) serve as a bioindicator species for trace metals pollution?

Bats are an endangered group of mammals that are very sensitive to environmental stresses. One of such stress factor is trace metals pollution which threatens populations of insectivorous bats due to their top position in the food webs and exceptionally long life span. In our research Pipistrellus kuhlii was tested as a promising indicator species (urban-dwelling, sedentary, with limited daily home-range) for trace metal exposure of bats. We measured concentrations of Pb, Cu, Zn and Cd in internal and external tissues of bats from the industrial city Mariupol and the village Karlovka, Ukraine, to answer the following questions: (1) Do metal concentrations in soil samples differ between urban and rural areas, and between populations of P. kuhlii from those areas? (2) Does metal contamination differ between individuals of different sexes and ages? (3) Whether fur and/or wing membrane, the two tissues that can be collected from live bats, can be used as proxies of metal contamination in internal tissues (liver, kidney, lung, forearm bones) of P. kuhlii? Metal concentrations in soil samples were significantly higher in the city. Bats from the city accumulated significantly more Cd, Pb and Zn in external tissues than those from the rural area. Females accumulated more Cd than males, and this-year-born did not differ significantly from adult individuals. We did not find, however, significant positive correlations between metal concentrations in external and internal tissues, indicating that external tissues cannot serve as an indicator of the metal contamination of internal tissues in P. kuhlii.

The dataset of bat (Mammalia, Chiroptera) occurrences in Ukraine collected by the Ukrainian Bat Rehabilitation Center (2011-2022).

Background: Bats are of high conservational status in most European countries. All bats are under legal protection in Ukraine and included in the national Red Data Book. However, bats remain one of the least studied groups of mammals in Ukraine. Their cryptic lifestyle limits the possibilities of direct observations and, as a result, data on bat distribution are incomplete. Wildlife rehabilitation centres accumulate a plethora of records of wild animals and those data may significantly contribute to knowledge on the species range, phenology and habitat preferences.This paper presents the data accumulated from over a decade of work by the Ukrainian Bat Rehabilitation Center (formerly The Bat Rehabilitation Center of Feldman Ecopark), the premier organisation engaged in the rescue and rehabilitation of bats across Ukraine. In addition to in-person data collected by Ukrainian Bat Rehabilitation Center staff, the Center also accepts observations of bat encounters from citizens. The Center's dataset boasts over 20,000 distinct observations, which are the subject of this paper. New information: This dataset, spanning 2011-2022, contains a total of 20,948 records of bat findings, 19,024 of which consist of records directly identified by UBRC team members. The remaining 1924 observations were provided by citizens through helpline. Data on 16 species and one subspecies have been collected. The highest number of records belongs to Nyctalusnoctula (n = 15889), followed by Eptesicusserotinus (n = 2017) and Pipistrelluskuhliilepidus (n = 2001). Less than 10% of these records have been previously published; the rest are presented in this paper for the first time. The dataset is particularly rich in information on bats in human settlements and is (to the best of the authors' knowledge) the largest dataset on bats within human-modified landscapes ever collected from the territory of Eastern Europe. The entire dataset is available through the Global Biodiversity Information Facility (GBIF).

First data on bacteria associated with bat ectoparasites collected in Kharkiv oblast, Northeastern Ukraine.

BACKGROUND: Bats (Mammalia: Chiroptera) serve as natural reservoirs for many zoonotic pathogens worldwide, including vector-borne pathogens. However, bat-associated parasitic arthropods and their microbiota are thus far not thoroughly described in many regions across the globe, nor is their role in the spillover of pathogens to other vertebrate species well understood. Basic epidemiological research is needed to disentangle the complex ecological interactions among bats, their specific ectoparasites and microorganisms they harbor. Some countries, such as Ukraine, are particularly data-deficient in this respect as the ectoparasitic fauna is poorly documented there and has never been screened for the presence of medically important microorganisms. Therefore, the aims of this study were to provide first data on this topic. METHODS: A total of 239 arthropod specimens were collected from bats. They belonged to several major groups of external parasites, including soft ticks, fleas, and nycteribiid flies from six chiropteran species in Northeastern Ukraine. The ectoparasites were individually screened for the presence of DNA of Rickettsia spp., Anaplasma/Ehrlichia spp., Bartonella spp., Borrelia spp., and Babesia spp. with conventional PCRs. Positive samples were amplified at several loci, sequenced for species identification, and subjected to phylogenetic analysis. RESULTS: Rickettsia DNA was detected exclusively in specimens of the soft tick, Carios vespertilionis (7 out of 43 or 16.3%). Sequencing and phylogenetic analysis revealed high similarity to sequences from Rickettsia parkeri and several other Rickettsia species. Bacteria from the family Anaplasmataceae were detected in all groups of the ectoparasites (51%, 122/239 samples), belonging to the genera Anaplasma, Ehrlichia, and Wolbachia. The detection of Bartonella spp. was successful only in fleas (Nycteridopsylla eusarca) and bat flies (Nycteribia koleantii, N. pedicularia), representing 12.1% (29/239) of the collected ectoparasites. No DNA of Babesia or Borrelia species was identified in the samples. CONCLUSIONS: We report for the first time in Ukraine the molecular detection of several bacterial agents in bat ectoparasites collected from six species of bats. The data presented extend the knowledge on the distribution of ectoparasite species in bats and their involvement in potentially circulating agents pathogenic for humans and vertebrate animals.

Blood Parasites and Health Status of Hibernating and Non-Hibernating Noctule Bats (Nyctalus noctula).

Co-existence of bats with a wide range of infectious agents relates to their co-evolutionary history and specific physiology. Here, we examined blood samples collected during hibernation and the post-hibernation period to assess the influence of trypanosomes and babesias on the health status of 50 Noctule bats (Nyctalus noctula) using nested PCR. The impact of blood parasites on health was assessed by analysis of haematology and blood chemistry parameters in 21 bats. Prevalence of trypanosomes (Trypanosoma dionisii and T. vespertilionis) and babesia (Babesia vesperuginis) was 44% and 8%, respectively. Analysis of blood parameters indicated impact of babesia on acid-base balance. Blood chemistry parameters showed a significant decrease in total dissolved carbon dioxide and bicarbonate, increased anion gap, and no change in blood pH, suggesting compensated metabolic acidosis. Adverse effects of babesia were only apparent in hibernating bats. Our results suggest differences in the pathogenicity of trypanosomes and babesia in bats. While trypanosomes in general had no significant impact on the health status, we observed alterations in the blood acid-base balance in Babesia-infected bats during hibernation. Despite being infected, Babesia-positive bats survived hibernation without showing any clinical signs.

Quantitative evaluation of individual food intake by insectivorous vespertilionid bats (Chiroptera, Vespertilionidae).

Insectivorous bats provide important ecosystem services, especially by suppressing and controlling the insects' biomass. To empirically quantify the number of insects consumed by European vespertilionid bats per night, we estimated their ratio of dry mass of feces to mass of consumed insects. This study combines the results of feeding in captivity and the data obtained in field surveys; dry mass of feces was measured in both cases. In captivity, we analyzed the effect of species, age and sex of bats, species of insects consumed and the mass of food portion on the dry mass of feces. Using coefficients of the regression model, we estimated the number of insects consumed by free-ranging bats based on dry mass of their feces. According to our estimates, on average, one individual of one of the largest European bat species, Nyctalusnoctula, consumes 2.2 g (ranging from 0.5 to 8.2 g) of insects per one feeding night, while the smallest European bats of genus Pipistrellus consume 0.4 g (ranging from 0.1 to 1.3 g), further confirming the importance of insectivorous bats for ecosystem services. This publication offers the novel method for the estimation of insects' biomass consumed by bats.

Wing membrane and Fur as indicators of metal exposure and contamination of internal tissues in bats.

All European bats are protected by the EU and Associated Members legal regulations. Being insectivorous and top predators, they can be particularly exposed to persistent organic and inorganic pollutants. It is surprising how little is known about the impact of environmental pollutants on bats from physiological to populational levels. In this study we focused on contamination with trace metals of first-year bats from Kharkiv city, NE Ukraine. Tissues from the carcasses of two species, Nyctalus noctula (n = 20) and Eptesicus serotinus (n = 20), were used for metal analysis. The samples of external (wing membrane, fur) and internal (liver, lung, kidney, bones) tissues were analysed for contents of Pb, Cu, Zn, and Cd to see whether fur or wing membrane can be used as proxies for metal contamination of the vital internal tissues. In E. serotinus, significant positive correlations in Pb concentrations were found between all external and internal tissues. For Cd only, correlation between the fur and lung was found, for Cu between the fur and liver, and for Zn between the fur and kidney. In contrast, for N. noctula, only one such correlation was found - between Zn concentrations in the fur and kidney. The tissues differed significantly in concentrations of all studied metals, with no difference between the species. The results showed that the fur and wing membrane can be used as good proxies for Pb concentrations in internal organs of E. serotinus, but not necessarily for other metals or for N. noctula. The results for Pb are, however, encouraging enough to conclude that the topic is worth further studies, covering more species, a wider age range and more diverse environments.

DNA metabarcoding and spatial modelling link diet diversification with distribution homogeneity in European bats.

Inferences of the interactions between species' ecological niches and spatial distribution have been historically based on simple metrics such as low-resolution dietary breadth and range size, which might have impeded the identification of meaningful links between niche features and spatial patterns. We analysed the relationship between dietary niche breadth and spatial distribution features of European bats, by combining continent-wide DNA metabarcoding of faecal samples with species distribution modelling. Our results show that while range size is not correlated with dietary features of bats, the homogeneity of the spatial distribution of species exhibits a strong correlation with dietary breadth. We also found that dietary breadth is correlated with bats' hunting flexibility. However, these two patterns only stand when the phylogenetic relations between prey are accounted for when measuring dietary breadth. Our results suggest that the capacity to exploit different prey types enables species to thrive in more distinct environments and therefore exhibit more homogeneous distributions within their ranges.

Multiple isotope tracers in fur keratin discriminate between mothers and offspring.

RATIONALE: Isoscape origin models for mammals may be impaired by fractionation and routing of isotopes during lactation. Here, we tested if the stable carbon, nitrogen and hydrogen ratios of juvenile bats differ from those of their mothers and if derived isotopic dietary niches and geographical assignments vary accordingly between mothers and juveniles. METHODS: During the post-lactation period, we collected fur of juvenile and female common noctule bats (Nyctalus noctula) from the same maternity roost. Using a combination of elemental analysis and stable isotope ratio mass spectrometry, we measured the hydrogen, carbon and nitrogen isotope ratios in fur keratin. The hydrogen isotope ratios were measured for the non-exchangeable portion of hydrogen in keratin. The derived isotopic niches and isoscape origin models were compared between mothers and juveniles. RESULTS: The fur keratin of juveniles was enriched by 1.6‰ in 15 N and depleted by 2.9‰ in 13 C compared with that of the mothers. In addition, the hydrogen isotope ratios were 13.4‰ lower in the fur keratin of juveniles than in that of mothers. The estimated isotopic niches of nursing females and juveniles were not overlapping and differed in size. Overall, the isoscape origin models projected juveniles as being from a more northern origin than the mothers; yet both models suggested the study site as a likely place of origin. CONCLUSIONS: Our results suggest that isotope ratio data of juvenile bats should not be used for transfer functions in isoscape origin models because of isotopic routing and discrimination during lactation. Not accounting for age effects may increase the inaccuracy of geographical assignments in mammals when based on stable hydrogen isotopes.

Diverse replication-associated protein encoding circular DNA viruses in guano samples of Central-Eastern European bats.

Circular replication-associated protein encoding single-stranded DNA (CRESS DNA) viruses are increasingly recognized worldwide in a variety of samples. Representative members include well-described veterinary pathogens with worldwide distribution, such as porcine circoviruses or beak and feather disease virus. In addition, numerous novel viruses belonging to the family Circoviridae with unverified pathogenic roles have been discovered in different human samples. Viruses of the family Genomoviridae have also been described as being highly abundant in different faecal and environmental samples, with case reports showing them to be suspected pathogens in human infections. In order to investigate the genetic diversity of these viruses in European bat populations, we tested guano samples from Georgia, Hungary, Romania, Serbia and Ukraine. This resulted in the detection of six novel members of the family Circoviridae and two novel members of the family Genomoviridae. Interestingly, a gemini-like virus, namely niminivirus, which was originally found in raw sewage samples in Nigeria, was also detected in our samples. We analyzed the nucleotide composition of members of the family Circoviridae to determine the possible host origins of these viruses. This study provides the first dataset on CRESS DNA viruses of European bats, and members of several novel viral species were discovered.

Strontium-90 and caesium-137 activity concentrations in bats in the Chernobyl exclusion zone.

Bats are a protected species and as such may be an object of protection in radiological assessments of the environment. However, there have previously been only few radioecological studies of species of bats. In this paper, results for >140 measurements of (90)Sr and (137)Cs in 10 species of bats collected within the Chernobyl zone are presented. There was some indication of a decreasing transfer of (90)Sr with increasing deposition, although this was inconsistent across species and explained little of the observed variability. There was no difference between male and female bats in the transfer (expressed as the ratio of whole-body activity concentrations to those in soil) of either radionuclide. There was considerable variability in transfer across all species groups. At two sites where there were sufficient data, Eptesicus serotinus was found to have higher transfer than other species.