New insights into the taxonomy and phylogeny of social voles inferred from mitochondrial cytochrome b sequences

We sequenced the entire cytochrome b gene in Microtus paradoxus from Turkmenistan and Microtus socialis from Crimea and Kalmykia. Phylogenetic relationships among social voles were reconstructed by the inclusion into analyses of a further 23 published haplotypes belonging to six species. The two probabilistic methods which were used in phylogenetic analyses, the Bayesian inference and Maximum Likelihood, yielded very similar results. Both trees showed two highly divergent lineages which were further subdivided into seven species. The socialis lineage encompassed four species (M. socialis, M. irani, M. anatolicus, and M. paradoxus), and the remaining three species clustered into the guentheri lineage (M. guentheri, M. hartingi, M. dogramacii). The ranges for nucleotide divergences between seven species of social voles (4.95–9.28% and 4.18–8.81% for mean and net divergences, respectively) mainly exceeded 4.3%, which is frequently regarded as the conservative cut-off between sibling species in the specious genus Microtus.     

Morphological variation of the five vole species of the genus Microtus (Mammalia,Rodentia, Arvicolinae) occurring in Greece

Morphometric data for the five vole species of the genus Microtus living in Greece are old, sparse, poor and insufficiently analysed. This work aims to give the first comprehensive morphometric analysis of body and skull inter‐ and intraspecific variation for M. (M.) guentheri, M. (M.) rossiaemeridionalis, M. (Terricola) subterraneus, M. (T.) felteni and M. (T.) thomasi, applying multivariate statistics to 28 linear morphometric variables. It was based on ample material (202 adult individuals) using samples from localities that adequately cover the entire distributional range of each species in Greece. The five species and the two subgenera (Microtus and Terricola) were morphometrically clearly distinguished and discriminating variables were revealed. However, morphometrics did not provide robust criteria to infer phylogenetic relations among species. Furthermore, three species, M. (M.) guentheri, M. (M.) rossiaemeridionalis and M. (T.) thomasi, exhibited considerable intraspecific size or shape variation, which was mostly random and not associated with geographical proximity. Comparisons with data in the literature, mainly concerning populations of these species from adjacent areas, indicate that the Greek M. (M.) guentheri, M. (M.) rossiaemeridionalis and M. (T.) thomasi tend to be smaller than their conspecifics, while M. (T.) subterraneus and M. (T.) felteni are about equal in size to their Balkan relatives.     

Diet of the Long-eared Owl,Asio otus,in Central Anatolia (Aves: Strigidae)

In a one-year study on the diet of the Long-eared Owl, Asio otus, in the area of Karap?nar in Central Anatolia, 5324 prey items were recovered from 3314 pellets collected at monthly intervals. At least 7 species of mammals belonging to Insectivora (Crocidura suaveolens) and Rodentia (Allactaga williamsi, Cricetulus migratorius, Meriones tristrami, Mescocricetus brandti, Microtus guentheri, M. sp., Mus macedonicus) were identified. Rodentia were dominant and contributed 93.8% of the number of prey items of the Long-eared Owl. Microtus guentheri accounted for 83.9% of the rodent remains. The diet pattern was more diverse in summer, when some 24% of the diet consisted of non-mammal prey items (birds, insects).     

Intraspecific epigenetic polymorphism of the East European vole (Microtus levis Miller, 1908) in South-eastern Europe and Turkey

This study aims to represent the first report on population variation of 20 non-metric skull characters in East European vole (Microtus levis) from the Balkan (populations from Northern Dobruja; Southern Dobruja; East part of the Danube Plain; North-east Trace; Sofia field; South-east Trace) and Anatolian peninsulas (populations from North-west Anatolia region and Central Anatolia region), on the basis of which to determine its epigenetic variability and to analyse their mutual geographical epigenetic relations through comparison of the epigenetic divergence among them. Estimation of epigenetic variation of the studied populations of M. levis showed similar pattern of variation, but it is mostly higher than the other rodent species with a similar range of distribution, such as Microtus arvalis, Mus musculus, Apodemus sylvaticus, Apodemus flavicollis and Clethrionomys glareolus. Each one of the studied traits manifested some polymorphism. Moreover, all the calculated epigenetic distances (MMD) were statistically insignificant (P < 0.05) and epigenetic cranial uniqueness (MU) of any studied population was not found. These results reveal lack of expressed geographic relationship of population epigenetic variability in East European vole. The revealed populations epigenetic polymorphism of M. levis gives an opportunity for more complete assessment of variability and biological diversity of this species, but further research is necessary to elucidate its population epigenetics, especially as the data obtained in recent investigations of cranial morphology of the sibling species from the group the M. arvalis (sensu lato) added new locations to the distribution map of the East European vole in Eurasia.     

Social but lonely: Species delimitation of social voles and the evolutionary history of the only Microtus species living in Africa

The social vole of Cyrenaica, Libya, is the only extant representative of the Cricetidae family found in Africa. Its taxonomic status has been under debate, partly due to the problematic systematics of the entire group of social voles and partly due to the lack of morphological and molecular data from Cyrenaican specimens. In this study, we applied ancient DNA protocols to produce three cytochrome b (cytb) sequences of Cyrenaican voles and built a phylogenetic reconstruction (195 sequences in total) incorporating all available cytb sequences of the remaining social voles, other representatives of the genus Microtus and closely related cricetids. We used this phylogeny to test the performance of the model-based, single-locus, species delimitation approach implemented in mPTP and delimited nine species of social voles. Among them are the Cyrenaican vole, Microtus mustersi and its sister species M. guentheri, distributed along the Mediterranean coasts of southwestern Asia. Biogeographical reconstruction of ancestral area and molecular clock estimations of the time since the divergence of the two sister lineages suggest that their common ancestor dispersed into Africa through a coastal route and was isolated in Cyrenaica as a result of population fragmentation associated with Middle Pleistocenic pluvial/interpluvial cycles. Geographic isolation triggered the speciation process, but species distribution modeling gave evidence of subsequent niche divergence; M. guentheri has adapted to the xeric conditions of its distributional area, while M. mustersi benefited from the milder Cyrenaican climate. The Cyrenaican vole is a relict species more than 200,000 years old, has a small and isolated distribution and probably merits conservation.     

mtDNA analysis reveals the ongoing speciation on Greek populations of Microtus (Terricola) thomasi (Arvicolidae,Rodentia)

The present article extends our previous work on the phylogenetic history of Microtus (Terricola) thomasi, analysing cytochrome b, 12S rRNA, 16S rRNA and the control region in 65 Greek populations. The analysis revealed three clades: one grouping the populations from Peloponnisos (Southern Greece); the second, the populations from Agios (Ag.) Stefanos and Evvoia island (Central East Greece); and the third, all the remaining populations with no geographical substructure. Genetic distances were low for most populations, with only the populations of Evvoia and Ag. Stefanos being substantially distant. Thus, although this species has a recent colonization history and probably descends from a highly polymorphic ancestor, a monophyletic and highly differentiated lineage is formed in Greece and is distributed in Ag. Stefanos and Evvoia. Molecular differentiation, distinct geographical distribution and restriction of gene flow between this lineage and the rest of the Greek populations provide evidence for its probable subspecific status, Microtus (Tericola) thomasi atticus. A possible mechanism leading the differentiation process of the proposed subspecies is suggested, based on the displacement of this species in central Greece by its congeneric, probably better‐fitted Microtus (Microtus) guentheri and the subsequent separation of Ag. Stefanos and Evvoia from the remaining Greek populations. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95 , 117–130.     

The Iranian vole Microtus irani occurs in Lebanon (Mammalia: Rodentia)

We studied 1140 bp cytochrome b sequences of social voles from three localities in Lebanon. The results were compared with published sequences representing seven species of social voles. New sequences from Lebanon clustered with reference samples of two species: M. guentheri and M. irani. While M. guentheri was already reported for Lebanon, M. irani is a new addition to the fauna of Lebanon, and the third known record for the species. Animals were collected in two localities above Tripolis at 855 m and 1430 m a.s.l., respectively.     

Chromosomal variation in social voles: a Robertsonian fusion in Günther’s vole

The study reports on chromosomes in several populations of social voles from south-eastern Europe and the Middle East. The standard karyotypes of individuals of Microtus hartingi and Microtus guentheri originating from both south-eastern Europe and Asia Minor comprised 54 mostly acrocentric chromosomes. However, variation between populations was found in the amount and distribution of C-heterochromatin in certain autosomes and the sex chromosomes. Furthermore, a specific pattern of argyrophilic nucleolar organizer region distribution was recorded in different geographic populations. In a population from Asia Minor, a heterozygous centric fusion of two autosomes was found. The G-banded karyotypes of M. guentheri and Microtus socialis were compared, and tandem fusions of autosomes were suggested as possible mechanism of the divergence. The karyotypes of the nine currently recognized species of social voles are reviewed, and implications of chromosomal data for systematics are evaluated.     

Eastern Beringian biogeography: historical and spatial genetic structure of singing voles in Alaska

Aim  Pleistocene climatic cycles have left marked signatures in the spatial and historical genetic structure of high‐latitude organisms. We examine the mitochondrial (cytochrome b) genetic structure of the singing vole, Microtus miurus (Rodentia: Cricetidae: Arvicolinae), a member of the Pleistocene Beringian fauna, and of the insular vole, Microtus abbreviatus, its putative sister species found only on the St Matthew Archipelago. We reconstruct the phylogenetic and phylogeographical structure of these taxa, characterize their geographical partitioning and date coalescent and cladogenetic events in these species. Finally, we compare the recovered results with the phylogenetic, coalescent and spatial genetic patterns of other eastern Beringian mammals and high‐latitude arvicoline rodents. Location  Continental Alaska (alpine and arctic tundra) and the St Matthew Archipelago (Bering Sea). Methods  We generated and analysed cytochrome b sequences of 97 singing and insular voles (M. miurus and M. abbreviatus) from Alaska. Deep evolutionary structure was inferred by phylogenetic analysis using parsimony, maximum likelihood and Bayesian approaches; the geographical structure of genetic diversity was assessed using analysis of molecular variance and network analysis; ages of cladogenetic and coalescent events were estimated using a relaxed molecular clock model with Bayesian approximation. Results  Regional nucleotide diversity in singing voles is higher than in other high‐latitude arvicoline species, but intra‐population diversity is within the observed range of values for arvicolines. Microtus abbreviatus specimens are phylogenetically nested within M. miurus. Molecular divergence date estimates indicate that current genetic diversity was formed in the last glacial (Wisconsinan) and previous interglacial (Sangamonian) periods, with the exception of a Middle Pleistocene split found between samples collected in the Wrangell Mountains region and all other singing vole samples. Main conclusions  High levels of phylogenetic and spatial structure are observed among analysed populations. This pattern is consistent with that expected for a taxon with a long history in Beringia. The spatial genetic structure of continental singing voles differs in its northern and southern ranges, possibly reflecting differences in habitat distribution between arctic and alpine tundra. Our phylogenetic results support the taxonomic inclusion of M. miurus in its senior synonym, M. abbreviatus.     

Cranial variation in Meriones tristrami (Rodentia: Muridae: Gerbillinae) and its morphological comparison with Meriones persicus,Meriones vinogradovi and Meriones libycus: a geometric morphometric study

Jirds (genus Meriones) comprise a group of rodents, of which the biodiversity is still poorly known. Reason for this is that several species of similar morphologies are known to occur sympatrically. In the north‐west of Iran, four such species occur: Meriones tristrami, Meriones persicus, Meriones vinogradovi and Meriones libycus, prone to several issues of taxonomical ambiguity. A proper characterization of morphological distinctiveness between these species, in relation to the variation within species, could provide the required information for species diagnosis and identification. As some cranial characters of M. tristrami, M. persicus and M. vinogradovi are quite similar, demarcations of species‐specific phenotypic variation have proven to be difficult. To tackle this problem, this study involves a geometric morphometric analysis of skull shape and size, incorporating a large representative sample of these four species, originating from most parts of their natural distribution range (especially for M. tristrami). It is first tested whether M. tristrami can be distinguished from the other sympatric species, and if so, to what degree the species shows a geoclimatic pattern in its skull shape and size when comparing different populations. The shape and size analyses show that M. libycus can be distinguished because of its largest skull and the relatively largest tympanic bulla, and that M. tristrami can be distinguished from the other species. At an intraspecific level in M. tristrami, the Iranian groups (Qazvin and west Iran) do not differ in shape among them, but do so in skull size. They could, however, be distinguished in skull shape from the non‐Iranian populations included (Turkey and Jordan). To what degree this continuous data can now be translated into discrete and diagnostic features, useful for taxonomic purposes, remains to be studied.     

Heterochromatin study and geographical distribution of Microtus species (Rodentia,Arvicolinae) from Greece

In a sample of 108 underground voles from 23 Greek localities, the species Microtus felteni, M. guentheri, M. rossiaemeridionalis and M. subterraneus were identified, based on external body morphology and karyotype. Moreover, the implemented C-banding staining technique revealed the heterochromatin distribution in the chromosomes of the above species. All M. guentheri (2n=54, FN=54) and M. rossiaemeridionalis (2n=54, FN=56) specimens displayed the typical karyotypes of these species, respectively. The M. subterraneus specimens belonged to the chromosomal race with 2n=52, FN=60 of this species, apart from a single individual that demonstrated a medium-sized, subtelocentric autosome in heterozygous condition (2n=52, FN=61). Furthermore, M. felteni individuals, trapped again after many years, were karyologically studied (2n=54, FN=56) and the C-banding pattern for this species is hereby presented for the first time. Finally, the study of meiotic preparations in M. guentheri and M. rossiaemeridionalis males verified the asynaptic behaviour of their sex chromosomes. The karyotype of the four studied Microtus species does not seem to have diversified much from the putative ancestral arvicoline karyotype (2n=56, FN=56). On the other hand, the heterochromatin accumulation in the sex chromosomes, particularly prominent in M. felteni and M. rossiaemeridionalis, represents this common tendency in several Microtus species.     

Aquatic adaptations in a Neotropical coral snake: A study of morphological convergence

Micrurus surinamensis is an aquatic member of the genus Micrurus. This species is known for its highly specialized venom and distinctive diet, mostly made of aquatic vertebrates. Here, we explore both external (head and body) and skull shape morphologies in M. surinamensis, comparing it with two terrestrial species of the genus (M. lemniscatus and M. spixii) and to aquatic and terrestrial species of distantly related groups. We use both traditional and geometric morphometrics to determine whether the presence of similar traits in head shape morphology is rather the result of adaptive convergences between M. surinamensis and other aquatic species, or whether it is the product of phylogenetic conservatism within the genus. Results from both traditional and geometric morphometrics show that M. surinamensis can be considered convergent with aquatic species, mainly in the skull shape. Micrurus surinamensis differs from the two terrestrial species of Micrurus by having a wider head, smaller distance between nostrils, and a long tail. Geometric morphometric analysis shows that despite having an extremely conserved skull and mandible shape, M. surinamensis shows a longer supratemporal and quadrate bones than in terrestrial Micrurus, indicating a larger gape for this species. A more kinetic skull combined with a larger gape would allow M. surinamensis to feed on fish, which represent larger and wider prey that contrast with the elongate prey, which compose the main diet of species in the genus Micrurus. Our results illustrate the importance of both phylogenetic conservatism and adaptation in shaping species morphology.     

Evolutionary changes in craniomandibular shape in the great cats (Neofelis Griffith and Panthera Oken)

Evolutionary shape changes in skull and mandibular anatomy was analysed in 223 specimens of pantherine felids (Neofelis nebulosa, Panthera leo, Panthera onca, Panthera pardus, Panthera tigris, Panthera uncia) compared to a small‐felid outgroup, consisting of 86 specimens of nine different species, using digital surface morphometry on 25 (skull) and 17 (mandible) landmarks. Shape evolution in the pantherine species is complex and nonlinear, and involves both large‐scale and small‐scale shape changes. Shape changes frequently differ among the ingroup species, but the four large Panthera species (leo, onca, pardus, tigris) bear some resemblance to each other. The leopard and jaguar bear the closest resemblance to each other, and several shape changes are common to the lion and tiger, but have probably evolved convergently as a result of large size. The lion has undergone the largest and most numerous shape changes from a small‐felid outgroup. Certain shape changes in the skull and, in some respects, the mandible of the clouded leopard bear resemblance to those in the four large Panthera species. The snow leopard is often regarded as the most primitive of the extant Panthera, and skull and mandibular shape changes often diverge markedly from those observed in the other five ingroup taxa; its overall skull shape is rather similar to the small‐felid outgroup. This indicates that the shape changes in the clouded leopard are convergent with those of the four large Panthera species. Landmark integration showed no significant correlation with molecular phylogeny, chiefly owing to the snow leopard being placed among the four large Panthera species. A traditional phylogenetic topology with the snow leopard as the basal‐most species of Panthera yielded a weak but nonsignificant phylogenetic signal. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95 , 766–778.     

Integrative taxonomy of the Italian pine voles,Microtus savii group (Cricetidae,Arvicolinae)

The Italian endemic voles belonging to the Microtus savii group (de Sélys‐Longchamps, 1838) include five controversial taxa, namely M. savii savii, M. s. niethammericus Contoli 2003 ; M. s. tolfetanus Contoli 2003 , M. s. nebrodensis (Minà‐Palumbo, 1868) and M. brachycercus (Lehamann, 1961). A previous mtDNA phylogenetic analysis revealed three main mtDNA lineages that do not correspond to the recognized taxa. Here, we perform a comprehensive univariate and multivariate analysis of craniodental measurements in 151 specimens from 54 geographic samples belonging to the three main phylogroups. Qualitative data on the skull of 126 specimens were also obtained from 54 localities. Moreover, we sequenced the nuclear DNA β‐fibrinogen intron 7 (β‐fibint7) of 23 specimens from 12 localities and Cyt b sequences of 11 new specimens from seven localities. The pattern of morphological and cytogenomic divergence does not corroborate the current taxonomic framework and indicates that M. nebrodensis is a Sicilian endemic species.     

Exploring phylogeography and species limits in the Altai vole (Rodentia: Cricetidae)

Natural hybridization between species is not a rare event. In arvicoline rodents, hybridization is known to occur in the wild and/or in captivity. In the Microtus arvalis group, cytogenetic studies revealed that there were two distinct chromosomal forms (2n = 46 but a different fundamental number of autosomes). These forms have been attributed to two cryptic species: the common (arvalis) and Altai (obscurus) voles. Recently, individuals with intermediate karyotypes (F₁ and backcrosses) were discovered in central European Russia, and, for this reason, other studies have regarded obscurus and arvalis as conspecific. In the present study, to address the question of the species limits in the Altai vole and to infer its evolutionary history, a phylogeographical analysis combined with multivariate morphometric methods and original chromosome data was performed. Two obscurus lineages were identified: the Sino‐Russian and South Caucasian lineages. Both lineages are characterized by low genetic diversity, resulting, in the former, from a past bottleneck event caused by encroaching periglacial areas and, in the latter, from recent rapid population divergence. Introgressive hybridization between the Altai and common voles appears to be the result of a secondary contact following the Last Glacial Maximum in central European Russia. Despite the fact that speciation is an ongoing process in most arvicoline species, the common and Altai voles are genetically divergent, morphologically and karyologically distinct, and exhibit contrasting evolutionary histories. For all these reasons, they should be ranked as species: M. arvalis and M. obscurus. © 2013 The Linnean Society of London     

mtDNA evidence for a local northern latitude Pleistocene refugium for the root vole (Microtus oeconomus,Arvicolinae, Rodentia) from Eastern Poland

We analysed the genetic structure of 33 populations of the root vole (tundra vole, Microtus oeconomus, Pallas, 1776) inhabiting their typical habitats, located at different distances from the southern boundary of the species’ range (52°14′–53°56′ N) in eastern Poland. We determined its phylogeographic pattern as well as the possible occurrence of a small, local high‐latitude refugium of this species in southern Poland, previously suggested in palaeontological studies. 908 bp of cytochrome b sequences were analysed from 439 root voles, and 21 mtDNA cytb haplotypes belonging to the Central European (CE) phylogroup were found. Haplotype diversity in the examined populations varied between 0 and 0.872 (mean: 0.425 ± 0.332), while nucleotide diversity ranged between 0 and 0.62% (mean: 0.235% ± 0.217). Within the CE phylogroup of M. oeconomus, we identified with high bootstrap support a newly separated group of M. oeconomus that evolved from CE, denoted CE‐PL S. This group is located in the southern and central part of eastern Poland and most likely diverged from phylogroup CE in a small, cryptic refugium situated in southern Poland, in the Kraków‐Cz?stochowa Upland and/or the Holy Cross Mountains during the LGM and Younger Dryas.     

Cranial shape variation in jacarean caimanines (Crocodylia,Alligatoroidea) and its implications in the taxonomic status of extinct species: The case of Melanosuchus fisheri

Melanosuchus niger (Crocodylia, Alligatoroidea) is one of the six living caimanine species widely distributed throughout the Amazon River basin today. Although there is only one extant species of Melanosuchus, fossil material assigned to this genus, represented by M. fisheri, has been reported from the late Miocene in South America. However, the validity of this taxon has been questioned and a recent investigation indicates that the referred specimen of M. fisheri (MCZ 4336) actually belongs to Globidentosuchus brachyrostris, while those diagnostic characters present in the holotype (MCNC 243) fall into the spectrum of intraspecific variation of M. niger. Here, we compare the skull shape of the holotype of M. fisheri with the ontogenetic series of the four jacarean species (M. niger, Caiman yacare, Caiman crocodilus, and Caiman latirostris) using 2D‐geometric morphometric analyses in two different views. The analyses indicate that MCNC 243 falls into the morphospace of M. niger and C. latirostris. Despite strong shape similarities between juveniles of C. latirostris and MCNC 243, further anatomical comparisons reveal notable differences between them. In contrast, no concrete anatomical differences can be found between MCNC 243 and M. niger, although shape analyses indicate that MCNC 243 is relatively robust for its size. Thus, this study is able to confirm that the genus Melanosuchus was present in the late Miocene, but it still remains unclear if MCNC 243 should be treated as a junior synonym or probably a sister species of M. niger. Its Miocene age favors the second option, but as the shape analyses were also not able to extract any diagnostic characters, it should be retained as Melanosuchus cf. niger.     

Geographic phenetic variation of two eastern-Mediterranean non-commensal mouse species, Mus macedonicus and M. cypriacus (Rodentia: Muridae) based on traditional and geometric approaches to morphometrics

We tested the hypothesis that skull shape within the genus Mus may vary with geographic location by assessing the extent and spatial distribution of phenotypic skull variation within and among two wild mouse species, M. macedonicus and M. cypriacus, using traditional and geometric morphometrics including a rather novel application of sliding semilandmarks. Shape was shown to be significantly correlated both with longitude and latitude in M. macedonicus, yet the correlation between morphometric and geographic distances was not significant, and morphometric differences between Asian and European populations were not higher than those within the particular continents. The phylogenetic signal was found to be stronger in dental characters than in cranial ones, however, overall concordance between the pattern of morphometric variation and the presumed history of M. macedonicus was rather weak. In both species, the dorsal and ventral sides of the skull were shown to covary in many aspects though there were also some differences between them, making the functional interpretation of these differences difficult. Discrimination between M. cypriacus and M. macedonicus as well as discrimination between two M. macedonicus subspecies was highly reliable using both traditional and geometric morphometric tools to analyze skull measurements.     

Sound communication in social voles (subgenus Sumeriomys)

ABSTRACT

The acoustic communication of three species of social voles from the subgenus Sumeriomys – Microtus socialis (two subspecies: M. s. socialis and M. s. goriensis), M. paradoxus and M. hartingi – are described. Vole sound communication includes two main signals: squeaks and singing. The sounds made by M. hartingi have significantly higher frequency parameters than those of other species. Voles of all species squeak in situations of distress, and the males sing during courtship of the females. However, singing in social voles is not a necessary pattern for sexual behaviour: less than half of M. s. socialis and M. paradoxus males sang, M. hartingi sang even more rarely and M. s. goriensis did not demonstrate this behaviour at all. Despite the great similarity of the squeaks, its parameters differ significantly between species and differ from those of the common voles. This introduces one more argument that M. paradoxus and M. socialis are independent species, as are the subgenera Sumeriomys and Microtus.