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We describe a population of pipistrelle-like bats from Príncipe Island (Gulf of Guinea, Western Central Africa) as a new species based on the molecular and morphological characteristics of six specimens collected more than 30 years ago. The description of this new species was not possible until the traditionally entangled systematics of the whole pipistrelle group was clarified in recent years with the inclusion of molecular techniques and adequate species sampling. In this new taxonomic framework, the new species was clearly included within the dark-winged group of the recently described genus Pseudoromicia. The pipistrelles from Príncipe Island present a moderately inflated skull in lateral view with inner upper incisors that are moderately bicuspids and a baculum distinctly long with expanded tips. Besides these morphological characters, the new bat species is distinguished by its dwarfism, being the smallest species recognized within the genus. The ecology and conservation status of this endemic island species are unknown and field studies are urgently needed to evaluate the situation and conservation threats to this new species in its natural habitat.

The Big Naked-backed Bat, Pteronotus gymnonotus, is one of the 15 species currently recognized of this genus, with relatively few specimens in scientific collections, besides being poorly studied. It has a geographical distribution spanning from México through Central America and reaching Perú and Brazil, in which it occupies a variety of habitats from desert to tropical forests below 400 meters above sea level. Here, we report the records that demonstrate its presence, and data about its natural history in southeastern México, the northernmost part of its geographic distribution range. Between June 2002 and July 2018, we captured specimens in 44 bat roosts located in southeastern México, including the Parque Estatal Agua Blanca, Macuspana, Tabasco; Grutas de Martínez de la Torre, Matías Romero Avendaño, Oaxaca; and in Cueva de Villa Luz, Tapijulapa, Tabasco. In the three locations mentioned, we recorded the occurrence of P. gymnonotus individuals, whose taxonomic identification at species level was corroborated by both morphological data and genetic analyses. Previously, the only records of P. gymnonotus in México were from only four specimens scattered across time, so these new recorded locations confirm the presence of this species in the country. In addition to this, in Agua Blanca State Park and Villa Luz Cave we found a reproductive resident population. The record from Grutas de Martínez de la Torre is located in the middle of the Tehuantepec Isthmus, a well known biogeographical barrier for many taxa in the transitional area to the Pacific lowland’s region. We suggest that the occurrence of P. gymnonotus in México is also associated with large remnants of evergreen and gallery forests, located in the lowland areas along the Gulf of México and in the north and east of the Tehuantepec Isthmus.

Two sibling bare-backed bat species (Pteronotus fulvus and P. gymnonotus) have been traditionally differentiated by their size. However, intermediate specimens between the two species have been found in sympatric populations along southern Mexico and it has been suggested that they may be the outcome of a hybridization process between the two species. We used one mitochondrial (COI), three nuclear markers (PRKCL, STAT5A and RAG2) and 13 microsatellites to explore the evolutionary relationships between these two species and elucidate whether the intermediate morphotypes correspond to hybrid individuals. These markers have been analyzed in sympatric and allopatric populations of the two species plus the closely related species Pteronotus davyi. We confirmed the species-level differentiation of the three lineages (P. fulvus, P. davyi and P. gymnonotus), but the phylogenetic hypotheses suggested by the nuclear and mitochondrial markers were discordant. We confirm that the discordance between markers is due to genetic introgression through the mitochondrial capture of P. fulvus in P. gymnonotus populations. Such introgression was found in all P. gymnonotus specimens across its sympatric distribution range (Mexico to Costa Rica) and is related to expansion/retraction species distribution pulses associated with changes in forest distribution during the Quaternary climate cycles. Microsatellite analyses showed contemporary genetic contact between the two sympatric species and 3.0% of the samples studied were identified as hybrids. In conclusion, we found a historical and asymmetric genetic introgression (through mitochondrial capture) of P. fulvus into P. gymnonotus in Mexico and Central America and a limited contemporary gene exchange between the two species. However, no relationship was found between hybridization and the intermediate-sized specimens from southern Mexico, which might likely result from a clinal variation with latitude. These results confirm the need for caution when using forearm size to identify these species in the field and when differentiating them in the laboratory based on mitochondrial DNA alone.

Conservation efforts for endangered animals commonly focus on the protection of foraging habitats, aiming to ensure sufficient food availability. However, the diet of many species is based on animals that undergo habitat shifts across ontogenetic life stages, yielding considerable differences between the lifelong habitat requirements of both predator and prey. Consequently, prey availability may not only depend on the suitability of the foraging grounds where predator and prey coincide, but also on habitats where the ecological requirements of the non-prey stages are fulfilled. In this study we test to what extent prey of the insectivorous bat Rhinolophus euryale (Blasius 1853) originate either from the grounds where they are consumed, or in areas/habitats outside the bat's foraging sites. We analyzed the diet of R. euryale , by identifying its prey to the species level using DNA metabarcoding, and by searching for its prey's larval feeding requirements in the literature. We found that the larvae of the moth prey grow both inside and outside the grounds where they are hunted by the bats once the moths reach their adult stage. The importance of prey that originated from outside the bat's foraging grounds varied considerably across seasons. As a result, R. euryale does not only rely on the landscape elements where it hunts, but also on other source areas/habitats that supply it with food. This study shows that conservation measures that aim to address the foraging requirements of predatory species should not be limited to merely protecting their foraging grounds, but should also take into account the ecological requirements of their prey throughout their life stages.

amplify prey's DNA. In this study we evaluated and compared the efficiency of two total DNA extraction methods and five primer pairs for the molecular identification of birds from scats, in particular from the giant noctule bat (Nyctalus lasiopterus). A modified DNA stool Mini Kit of Qiagen was tested against a modified silica method with a guanidinium thiocianate (GuSCN) applied after freezing and pulverizing the samples. We also checked two published vertebrate- and bird-generalist primer pairs and three bird-specific primer pairs designed by us (two pairs targeting the cytochrome b and one the cytochrome oxidase subunit I genes) that amplified shorter DNA fragments. The results show that pulverizing the scat remains before extraction was a very important step, presumably facilitating access to the well preserved DNA located inside the rachis of the feathers. The combination of our bird-specific designed primers showed a higher amplification rate than the generalist primers and allowed successful bird identification from the feathers excreted by the giant noctule bat in all the scat samples analyzed, independent of the preservation method used (dried and frozen). These methodological improvements will allow not only the study of the avian diet composition of the enigmatic giant noctule, but the extension of this methodology to other bird predators such as raptors.

Genetic divergence in bat communities was assessed on both sides of the Straits of Gibraltar and cryptic diversity was examined. Screening was carried out using partial sequences of the mitochondrial (mt)DNA cytochrome b gene on 399 individual bats belonging to the 18 species found on both sides of the Straits of Gibraltar. For those bats that showed important genetic discontinuities, molecular markers (ND1 and nuclear RAG2 genes) were added to expand the sampling process. Phylogenetic reconstructions were obtained using maximum parsinomy, genetic distances, maximum likelihood, and Bayesian criteria. As an estimate of bats’ flight performance, we measured for each species the wing aspect ratio and wing loading indexes, and correlated them with the maximum pairwise genetic distances obtained between southern Iberian and northern Moroccan populations. Genetic mtDNA distances between populations on both continents exceed 5% in seven out of 18 bat species analysed and unknown lineages were uncovered within the species complexes Myotis nattereri and Myotis mystacinus. We did not find a general pattern in the degree of permeability of the Straits of Gibraltar for bats. Genetic distances were not correlated with the ability to cross the Straits. Our study shows that the cryptic diversity uncovered among bats continues to increase as more regions are studied. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 96, 434–450. ADDITIONAL KEYWORDS: community – cryptic diversity – cytochrome b – dispersal – genetic differentiation – mitochondrial DNA – ND1 – RAG2 – phylogeography – Pleistocene.

Recently, several species of aerial-hawking bats have been found to prey on migrating songbirds, but details on this behaviour and its relevance for bird migration are still unclear. We sequenced avian DNA in feather-containing scats of the bird-feeding bat Nyctalus lasiopterus from Spain collected during bird migration seasons. We found very high prey diversity, with 31 bird species from eight families of Passeriformes, almost all of which were nocturnally flying sub-Saharan migrants. Moreover, species using tree hollows or nest boxes in the study area during migration periods were not present in the bats’ diet, indicating that birds are solely captured on the wing during night-time passage. Additional to a generalist feeding strategy, we found that bats selected medium-sized bird species, thereby assumingly optimizing their energetic cost-benefit balance and injury risk. Surprisingly, bats preyed upon birds half their own body mass. This shows that the 5% prey to predator body mass ratio traditionally assumed for aerial hunting bats does not apply to this hunting strategy or even underestimates these animals’ behavioural and mechanical abilities. Considering the bats’ generalist feeding strategy and their large prey size range, we suggest that nocturnal bat predation may have influenced the evolution of bird migration strategies and behaviour.

In a survey of bats from La Rioja (Spain), several specimens of the mystacinus group were captured at different mountain localities. Genetic and morphologic analyses have revealed the presence of two lineages within this group in La Rioja. The lineages have been identified as corresponding to two different species: Myotis mystacinus sensu stricto and the recently described M. alcathoe. Both species were found using the same nocturnal refugia (caves) and the same forest habitats. This study extends the distribution of M. alcathoe west and southwards and adds a new mammal species to the Iberian fauna.

Integrative taxonomy aims to document biodiversity by incorporating all useful characters to increase confidence in hypotheses about phylogenetic relationships. In this study, we combine data obtained independently from morphology, two maternally inherited mtDNA genes and two biparentally inherited nuDNA genes to make phylogenetic and taxonomic hypotheses about the Palaearctic members of the bat genus Eptesicus (Vespertilionidae). This genus is distributed worldwide (except for Antarctica) and is highly diversified, presenting one of the most entangled taxonomic puzzles among all mammals. Our results support restoring the genus Rhyneptesicus and separating E. isabellinus and E. pachyomus from E. serotinus and E. ognevi and E. anatolicus from E. bottae. Differences in the phylogenetic hypotheses from mtDNA and nuDNA data suggest the occurrence within E. serotinus of evolutionary processes such as mtDNA capture and secondary contacts between partially differentiated ecomorphs. These two evolutionary processes deserve more in-depth studies within the group

A cost-effective, reliable and efficient method of obtaining DNA samples is essential in large-scale genetic analyses. This study examines the possibility of using a threatened vole species, Microtus cabrerae, as a model for the collection and preservation of faecal samples for subsequent DNA extraction with a protocol based on the HotSHOT technique. Through the examination of the probability of multi-copies (mitochondrial) and single copy (microsatellite) loci amplification (including the genotype error) and of the DNA yield (estimated by real-time qPCR), the new protocol was compared with both the frequently employed methods that successfully use ethanol to preserve faecal samples and with commercial kit-based DNA extraction. The single-tube HotSHOT-based protocol is a user-friendly, non-polluting, time-saving and inexpensive method of faeces sample collection, preservation and PCR- quality gDNA preparation. This technique therefore provides researchers with a new approach that can be employed in high-throughput, noninvasive genetic analyses of wild animal populations

We investigate the contribution of the Iberian bat fauna to the cryptic diversity in Europe using mitochondrial (cytb and ND1) and nuclear (RAG2) DNA sequences. For each of the 28 bat species known for Iberia, samples covering a wide geographic range within Spain were compared to samples from the rest of Europe. In this general screening, almost 20% of the Iberian species showed important mitochondrial discontinuities (K2P distance values > 5%) either within the Iberian or between Iberian and other European samples. Within Eptesicus serotinus and Myotis nattereri, levels of genetic divergence between lineages exceeded 16%, indicating that these taxa represent a complex of several biological species. Other well-differentiated lineages (K2P distances between 5–10%) appeared within Hypsugo savii, Pipistrellus kuhlii and Plecotus auritus, suggesting the existence of further cryptic diversity. Most unsuspected lineages seem restricted to Iberia, although two have crossed the Pyrenees to reach, at least, Switzerland

Bat predation on birds is a very rare phenomenon in nature. Most documented reports of bird-eating bats refer to tropical bats that occasionally capture resting birds. Millions of small birds concen- trate and cross over the world’s temperate regions during migra- tion, mainly at night, but no nocturnal predators are known to benefit from this enormous food resource. An analysis of 14,000 fecal pellets of the greater noctule bat (Nyctalus lasiopterus) reveals that this species captures and eats large numbers of migrating passerines, making it the only bat species so far known that regularly preys on birds. The echolocation characteristics and wing morphology of this species strongly suggest that it captures birds in flight

The Lesser Antilles is a biodiversity hot spot but unfortunately human disturbance has taken its toll, causing dramatic population declines and even extinction of numerous endemic species. Nevertheless, today the rediscovery of previously thought extinct species is not uncommon. Often, old museum specimens and their original descriptions are the only information available for such species. The application of molecular phylogenetic relationships to extant species can help to elucidate pivotal information on their ecology and conservation. Erythrolamprus cursor is possibly an extinct colubrid racer from Martinique, currently classified as critically endangered on the International Union for Conservation of Nature Red List. Mitochondrial deoxyribonucleic acid sequences were obtained from four E. cursor specimens from the Muséum national d'Histoire naturelle (Paris) collections. All sequences recovered the same haplotype and the level of divergence between E. cursor and E. juliae, from the nearby island of Dominica, was lower than between other intraspecific distances within other Erythrolamprus. Furthermore, phylogenetic analyses confirm that these two species are sister taxa and share most recent common ancestry. We discuss that published ecological data available for the sister species (E. juliae) may help to elucidate information on this species' natural history, ultimately having important implications for a future conservation management program if E. cursor is to be found. We emphasize the urgent need to conduct an exhaustive survey on the supposed last population of E. cursor at Diamond Rock to establish the survival of this species there, to understand how it may have adapted to such an ecosystem, especially in sympatry of several introduced rodent species

In July 2002, several bats of the genus Plecotus (Geoffroy, 1818) were captured at two localities of ‘Ordesa y Monte Perdido’ National Park (Central Pyrenees, Spain). They showed external characters that appeared intermediate between those of P. auritus and P. austriacus. Morphometric and genetic analysis have revealed that these long-eared bats should be ascribed to the recently recognized species P. macrobullaris. This study extends the distribution of this new species, whose western limit was considered, until now, to lie in the Alps, and adds a new mammal species to the Iberian fauna.