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Two metrically defined subspecies have traditionally been recognised within the Palaearctic distribution range of the Egyptian fruit bat Rousettus aegyptiacus ;t he largerR. a. aegyptiacus in Egypt and the northern part of the Middle East and the smaller R. a. arabicus in the southern Middle East. An extensive material of R. aegyptiacus from all parts of this area, i.e. the Levant (incl. Turkey and Cyprus), Egypt (incl. Sinai), northern Sudan, Yemen, Oman, Iran, and Pakistan, as well as comparative samples from its sub-Saharan range, were tested using both morphological and genetic approaches in order to revise the species' taxonomic status. The results indicated two possible processes, depending on the method used. Genetic analysis of the mitochondrial genome (nd1 and cytb) indicated low variation (< 2.0% of genetic distance) and lack of geographical structure while morphometric analysis indicated significant metric differences. Two basic size morphotypes were found within the Palaearctic range, with a rather mosaic-like geographical distribution and a lack of clear size distinction between the two categories, though intermediate types were detected. Thus, we suggest that all Palaearctic populations of R. aegyptiacus represent one form, the nominotypical subspecies, which is uniform in genetic traits but plastic in metric traits.

We used the mitochondrial control region and nuclear microsatellites to assess the distribution patterns, population structure, demography and landscape genetics for the hedgehogs Erinaceus europaeus and Erinaceus roumanicus in a transect of the mid-European zone of sympatry. E. roumanicus was less frequent and restricted to regions with lower altitudes. Demographic analyses suggested recent population growth in this species. A comparison of patterns in the spatial variability of mitochondrial and nuclear DNA indicated less sex-biased dispersal and higher levels of gene flow in E. roumanicus. No evidence of recent hybridisation or introgression was detected. We interpreted these results by comparing with phylogeographic and palaeontological studies as well as with the occurrence of hybridisation in the Russian contact zone. We propose that Central Europe was colonised by E. roumanicus by the beginning of the Neolithic period and that there was a subsequent reinforcement stage as well as the formation of a zone of sympatry after the complete reproductive isolation of both species.

Aim The Mediterranean Basin is a centre of radiation for numerous species groups. To increase our understanding of the mechanisms underlying speciation and radiation events in this region, we assessed the phenotypic variability within the Pipistrellus pipistrellus–pygmaeus–hanaki species complex. Although bats form the second largest mammalian order, studies of insular evolution in this group are scarce. We approached this problem from a microevolutionary perspective and tested for the recurrence of the insular syndrome. Location The Mediterranean Basin, with a special focus on isolated populations from Corsica, the Maghreb, Cyprus, Cyrenaica and Crete. Methods Phenotypic variability was assessed by cranial morphometrics using the coordinates of 41 3D landmarks and associated geometric-morphometric methods. We analysed 125 specimens representing all of the lineages in the species complex. Differences between taxa and between insular and continental populations in cranial size, shape, form and allometries were tested using analyses of variance and visualized using boxplots and canonical variate analysis. Relationships between molecular data from a previous study (cytochrome b sequences) and morphometric data were tested with co-inertia analyses (RV test) and multivariate regressions. Results The three species were relatively well differentiated in cranial size and shape, and each species showed a significant amount of inter-population variability. Comparisons of pairs of insular versus continental populations revealed heterogeneities in cranial patterns among island phenotypes, suggesting no recurrent insular syndrome. Molecular and phenotypic traits were correlated, except for molecular and lateral cranium shape. Main conclusions The Pipistrellus pipistrellus–pygmaeus–hanaki species complex exhibits phenotypic variability as a result of the fragmentation of its distribution (especially on islands), its phylogenetic and phylogeographic history and, most probably, other evolutionary factors that were not investigated in this study. We found no recurrent pattern of evolution on islands, indicating that site-specific factors play a prevailing role on Mediterranean islands. The correlation between molecular and phenotypic data is incomplete, suggesting that factors other than phylogenetic relationships, potentially connected with feeding ecology, have played a role in shaping cranial morphology in this species complex.

Here, we present a study of the Pipistrellus pipistrellus species complex, a highly diversified bat group with a radiation centre in the Mediterranean biodiversity hotspot. The study sample comprised 583 animals from 118 localities representatively covering the bats' range in the western Palearctic. We used fast-evolving markers (the mitochondrial D-loop sequence and 11 nuclear microsatellites) to describe the phylogeography, demography and population structure of this model taxon and address details of its diversification. The overall pattern within this group includes a mosaic of phylogenetically basal, often morphologically distant, relatively small and mostly allopatric demes in the Mediterranean Basin, as well as two sympatric sibling species in the large continental part of the range. The southern populations exhibit constant size, whereas northern populations show a demographic trend of growth associated with range expansion during the Pleistocene climate oscillations. There is evidence of isolation by distance and female philopatry in P. pipistrellus sensu stricto. Although the northern populations are reproductively isolated, we detected introgression events among several Mediterranean lineages. This pattern implies incomplete establishment of reproductive isolating mechanisms in these populations as well as the existence of a past reinforcement stage in the continental siblings. The occurrence of reticulations in the radiation centre among morphologically and ecologically derived relict demes suggests that adaptive unequal gene exchange within hybridizing populations could play a role in speciation and adaptive radiation within this group.

Between 2001 and 2008, we recorded Myotis alcathoe at nine sites within three distant areas in the Czech Republic. The species identification was confirmed with cyt b sequences and four distinct haplotypes were identified. All the localities exhibit surprisingly uniform habitat characteristics: (1) old full-grown oak-hornbeam forests, with (2) numerous large trees in advanced stages of decay are present, and (3) a very small to large water bodies and/or patches of riparian vegetation surrounded by the forest. Using radiotracking techniques, we discovered 27 day roosts of M. alcathoe, located mostly in big oak, birch and lime trees inside extensive forest stands. All roosts were fissures or small cavities in a tree trunk and in branches in the canopies, some 16m above the ground. Bats preferred trees that were higher, had higher canopy and canopy basement and had larger diameter at breast height than other available trees. Roost trees were surrounded by lower trees with lower canopy basements than availabl...

The demographic history of Rhinolophus hipposideros (lesser horseshoe bat) was recon- structed across its European, North African and Middle-Eastern distribution prior to, during and following the most recent glaciations by generating and analysing a multi- marker data set. This data set consisted of an X-linked nuclear intron (Bgn; 543 bp), mitochondrial DNA(cytb-tRNA-control region; 1630 bp) and eight variable microsatel- lite loci for up to 373 individuals from 86 localities. Using this data set of diverse markers, it was possible to determine the species’ demography at three temporal stages. Nuclear intron data revealed early colonization into Europe from the east, which pre-dates the Quaternary glaciations. The mtDNA data supported multiple glacial refugia across the Mediterranean, the largest of which were found in the Ibero- Maghreb region and an eastern location (Anatolia/Middle East)–that were used by R. hipposideros during the most recent glacial cycles. Finally, microsatellites provided the most recent information on these species’ movements since the Last Glacial Maxi- mumand suggested that lineages that had diverged into glacial refugia, such as in the Ibero-Maghreb region, have remained isolated. These findings should be used to inform future conservation management strategies for R. hipposideros and show the power of using a multimarker data set for phylogeographic studies.

The distribution of pipistrelles of the Pipistrellus pipistrellus complex (= P. pipistrellus s.l.) reaches only marginally the African continent. These bats are known only from a narrow belt of the Mediterranean zone in Maghreb and from NE Libya. We analysed museum specimens of African populations of P. pipistrellus s.l. using both morphologic and genetic techniques and compared them with Eurasian specimens of the complex. The African representatives of P. pipistrellus complex include two morphologically, genetically and geographically distinct populations. One of them inhabits the Mediterranean part of Cyrenaica, Libya. Belonging to the P. pygmaeus genetic lineage, these bats are represented by larger and more rusty coloured individuals with large massive rostrum and canines. In morphologic traits, this population differs significantly from all Western Palaearctic populations of the P. pipistrellus complex. These bats differ by about 6–7% in genetic distance from P. pygmaeus s. str. Within the P. pygmaeus lineage Libyan bats seem to be unique in their echolocation calls: the maximum energy of terminal frequencies was recorded at about 45 kHz. We consider the Libyan pipistrelles to represent a separate species, Pipistrellus hanaki sp. nov. Another distinct African pipistrelle population inhabits the Mediterranean parts of NW African countries, Morocco, Algeria and Tunisia. Individuals from the latter population are small and somewhat darker members of the P. pipistrellus genetic lineage, with relatively short and narrow mesial part of rostrum. Although both morphological and genetic differences between this population and Eurasian P. pipistrellus s. str. were found (genetic distance about 3–5%), they are probably not sufficient for the separation of this form at the specific level. However, the differences from European samples show rather not a cline character and therefore potential subspecific level of NW African P. pipistrellus has to be taken into consideration.

Craseonycteris thonglongyai (Chiroptera: Craseonycteridae), an enigmatic taxon which shares morphological traits with both Rhinopomatidae and Emballonuridae was for the first time investigated with the aid of molecular phylogenetic techniques. Three methods of phylogenetic inference, parsimony, maximum-likelihood, and Bayesian phylogenetics were used. Based on 402 bp of DNA sequence from the mitochondrial cytochrome b gene, placement of Craseonycteridae within the superfamily Rhinolophoidea was demonstrated. Our results also suggest close proximity of Craseonycteridae to Hipposideridae rather than to Rhinopomatidae, close relationships between Megadermatidae and Rhinolophidae, sister group position of Pteropodidae to Rhinolophoidea, and closer affiliation of Nycteridae with the infraorder Yangochiroptera. Spectral analysis was in agreement with all these outcomes except for closer relationships of Craseonycteris with Rhinopomatidae.

Introduction The Vespertilionidae is the largest family of bats, characterized by high occurrence of morphologically convergent groups, which impedes the study of their evolutionary history. The situation is even more complicated in the tropics, where certain regions remain under-sampled. Results Two hundred and thirteen vespertilionid bats from Senegal (West Africa) were studied with the use of non-differentially stained karyotypes and multi-locus sequence data analysed with maximum likelihood and Bayesian methods. These bats were identified as 10 different taxa, five of which were distinctive from their nominate species (Pipistrellus hesperidus, Nycticeinops schlieffenii, Scotoecus hirundo, Neoromicia nana and N. somalica), based on both karyotypes and molecular data. These five cryptic taxa are unrelated, suggesting that these West African populations have long been isolated from other African regions. Additionally, we phylogenetically analysed 166 vespertilionid taxa from localities worldwide using GenBank data (some 80% of the genera of the family) and 14 representatives of closely related groups, together with our Senegalese specimens. The systematic position of several taxa differed from previous studies and the tribes Pipistrellini and Vespertilionini were redefined. The African Pipistrellus rueppellii was basal to the Pipistrellus/Nyctalus clade and the Oriental species Glischropus tylopus was basal to the East Asian pipistrelles within the tribe Pipistrellini. The African genus Neoromicia was confirmed to be diphyletic. Based on GenBank data, Eptesicus was polyphyletic, with the Asian E. nasutus and E. dimissus both supported as phylogenetically distinct from the Eptesicus clade. The subfamily Scotophilinae was confirmed as one of the basal branches of Vespertilionidae. Conclusions New taxa and new systematic arrangements show that there is still much to resolve in the vespertilionids and that West Africa is a biogeographic hotspot with more diversity to be discovered.

Background The Rhinopomatidae, traditionally considered to be one of the most ancient chiropteran clades, remains one of the least known groups of Rhinolophoidea. No relevant fossil record is available for this family. Whereas there have been extensive radiations in related families Rhinolophidae and Hipposideridae, there are only a few species in the Rhinopomatidae and their phylogenetic relationship and status are not fully understood. Results Here we present (a) a phylogenetic analysis based on a partial cytochrome b sequence, (b) new fossils from the Upper Miocene site Elaiochoria 2 (Chalkidiki, Greece), which represents the first appearance datum of the family based on the fossil record, and (c) discussion of the phylogeographic patterns in both molecular and morphological traits. We found deep divergences in the Rhinopoma hardwickii lineage, suggesting that the allopatric populations in (i) Iran and (ii) North Africa and the Middle East should have separate species status. The latter species (R. cystops) exhibits a shallow pattern of isolation by distance (separating the Middle East and the African populations) that contrasts with the pattern of geographic variation in the morphometrical traits. A deep genetic gap was also found in Rhinopoma muscatellum (Iran vs. Yemen). We found only minute genetic distance between R. microphyllum from the Levant and India, which fails to support the sub/species distinctness of the Indian form (R. microphyllum kinneari). Conclusion The mtDNA survey provided phylogenetic tree of the family Rhinopomatidae for the first time and revealed an unexpected diversification of the group both within R. hardwickii and R. muscatellum morphospecies. The paleobiogeographic scenario compiled in respect to molecular clock data suggests that the family originated in the region south of the Eocene Western Tethyan seaway or in India, and extended its range during the Early Miocene. The fossil record suggests a Miocene spread into the Mediterranean region, followed by a post-Miocene retreat. Morphological analysis compared with genetic data indicates considerable phenotypic plasticity in this group.

Previous genetic analyses have demonstrated that two phonic types of one of the most common European bats, the Common pipistrelle, belong to distinct species, although they are almost identical morphologically (45 kHz Pipistrellus pipistrellus and 55 kHz Pipistrellus pygmaeus). To reconstruct the history of the species complex and explain the codistribution of both forms in Europe and the Mediterranean, we performed phylogenetic analysis based on a 402-bp portion of the cytochrome b gene. Particular attention was paid to the eastern and southern parts of the range where no data were available. We found further distinctive allopatric haplotypes from Libya and Morocco. The difference of about 6-7% described in the Libyan population suggests the occurrence of a new species in the southern Mediterranean. The species status of Moroccan population is also discussed. The phylogeographic patterns obtained and analysis of fossil records support the hypothesis of expansion of both species into Europe from the Mediterranean region during the Holocene. The allopatric speciation model fits our data best. The paleobiographic scenario envisaged is corroborated also by molecular clock estimations and correlations with Late Neogene environmental changes in the Mediterranean region which ended with the Messinian salinity crisis.

The Malaysian Giant Turtle (Orlitia borneensis) is a poorly known turtle with rapidly decreasing numbers in nature in spite of its strong protection on paper. Most individuals of this species kept in European zoos and included in captive breeding programs are confiscates from the illegal trade for food consumption and their geographic provenance is unknown. This study was aimed to assess genetic and phenotypic variation of the founders of this captive population. We sequenced the mitochondrial cytochrome b gene and found 23 haplotypes. We constructed a haplotype network and examined demographic changes by Bayesian skyline plots of the effective population size. The maximum sequence divergence was less than 1.5% and the phylogenetic structure of the haplotypes was supported poorly. A close genetic similarity among sampled turtles was further confirmed by sequencing the nuclear R35 gene, while the geometric morphometrics of the shell-shape were likewise similar. Thus, the examined captive population of O. borneensis may be further treated as a single conservation unit.

Analyses of museum specimens can help illuminate temporal changes in wildlife genetics and distributions, and the objective of our study was to evaluate the suitability of skin samples from the past century for genomic analyses. We examined two European species with extensive genomic resources and existing data: the wild boar (Sus scrofa) and the wolf (Canis lupus). Populations of both species declined during the twentieth century, followed by a strong recovery. Moreover, Carpathian Mountain wolves are genetically divergent from Northern European lowland wolves, and their evolutionary history is incompletely understood. We analysed genetic variation from skins of 14 European and Near Eastern wild boars (1903–1948) and 18 Carpathian and Northern wolves (1938–1998). Samples were genotyped in duplicates with standard Illumina single nucleotide polymorphism (SNP) arrays (wild boar 60 K and wolves 170 K), and we retained SNPs with a genotyping rate of > 90%. We obtained a final set of 1595 SNPs for six wild boars (genotyping rate 0.99) and 1806 SNPs for 18 wolves (genotyping rate 0.96). We merged the best performing duplicate with modern data and calculated polymorphism (P), observed (HO) and expected (HE) heterozygosity per population. Our findings underline the difficulties in obtaining quality genomic profiles from museum skins, though a limited number of SNPs provided insights for future research. Wild boars exhibited lower polymorphism in historical than modern samples. In contrast, patterns in wolves appeared spatial rather than temporal, with higher P, HO and HE observed in Carpathian than Northern wolves across historical and modern samples.