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.     

Mitochondrial DNA and nuclear microsatellites reveal high diversity and genetic structure in an avian top predator,the white‐tailed sea eagle,in central Europe

We analysed 123 white‐tailed sea eagles (Haliaeetus albicilla) from (primarily central) Europe with respect to variability and differentiation based on 499 bp of the mitochondrial control region and genotypes at seven unlinked nuclear microsatellites. Variability was high (overall expected heterozygosity, haplotype and nucleotide diversity being 0.70, 0.764 and 0.00698, respectively) and both marker systems showed a subdivision into two main genetic clusters (microsatellites) or haplogroups (mtDNA). In line with earlier analyses focusing on populations from northern and eastern Europe, as well as from Asia, we found a high level of admixture in Europe and no signs of a bottleneck – despite a severe decline of white‐tailed sea eagle populations during the 20^th century. Europe is thus a global stronghold for this species not only with respect to the number of breeding pairs but also regarding the proportion of species‐wide genetic diversity. Our dense sampling revealed a possibly clinal variation within central Europe from north‐west to south‐east that was reflected by the distribution of mtDNA haplotypes as well as the two microsatellite‐based clusters. This population differentiation in central Europe probably originated from a geographically structured postglacial colonization and was later enhanced by recent demographic fluctuations. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 727–737.     

Genetic divergence and phylogeography in the genus <Emphasis Type="Italic">Nyctalus</Emphasis> (Mammalia,Chiroptera): implications for population history of the insular bat <Emphasis Type="Italic">Nyctalus azoreum</Emphasis>

We used three mitochondrial DNA fragments with different substitution rates (ND1, Cyt b and the CR) to infer phylogenetic relationships among six species of the genus Nyctalus, and compare levels of genetic divergence between the insular, vulnerable Nyctalus azoreum and its continental counterpart to assess the origins of the Azorean bat. The larger species found throughout the Palaearctic region (N. lasiopterus, N. aviator and N. noctula) share a unique chromosome formula (2n = 42) and form a monophyletic clade in our reconstructions. Nyctalus plancyi (= velutinus), a Chinese taxon with 2n = 36 chromosomes, is sometimes included in N. noctula, but is genetically very divergent from the latter and deserves full species status. All Cyt b and CR haplotypes of N. azoreum are closely related and only found in the Azores archipelago, but when compared to continental sequences of N. leisleri, levels of mtDNA divergence are unusually low for mammalian species. This contrasts with the high level of differentiation that N. azoreum has attained in its morphology, ecology, and echolocation calls, suggesting a recent split followed by fast evolutionary change. The molecular data suggest that N. azoreum originated from a European population of N. leisleri, and that the colonisation of the Azores occurred at the end of the Pleistocene. The Madeiran populations of N. leisleri also appear to have a European origin, whereas those of the Canary Islands probably came from North Africa. In spite of its recent origin and low genetic divergence, the Azorean bat is well differentiated and consequently represents a unique evolutionary unit with great conservation value.     

Historical demography of freshwater mussels (Bivalvia: Unionidae): genetic evidence for population expansion and contraction during the late Pleistocene and Holocene

Genetic variation was examined in two endangered mussel species, Epioblasma brevidens and Epioblasma capsaeformis, and in a non‐listed species, Lampsilis fasciola, in the Clinch River, Tennessee, USA, by screening mitochondrial DNA (mtDNA) sequences and nuclear DNA microsatellites. Patterns of mtDNA polymorphism exhibited different trends in long‐term population sizes for each species during the late Pleistocene and Holocene (～20 000 ya to present); namely, E. brevidens has declined over time, E. capsaeformis has remained demographically stable, and L. fasciola has expanded. However, analyses using microsatellites did not exhibit similar trends, perhaps because homoplasy had eliminated long‐term population signatures for the loci examined. For both marker types, long‐term effective population size (N_e) was low in E. brevidens, intermediate in E. capsaeformis, and high in L. fasciola. Moderately diverged mtDNA lineages, perhaps indicative of secondary contact, were observed in E. brevidens and E. capsaeformis. Perhaps the most surprising result of this study was the high level of genetic variation observed at both mtDNA and microsatellite DNA markers for L. fasciola, variation seemingly contrary to the relatively small demes that currently reside in the Clinch River. However, the data are consistent with known demographic and life‐history traits of these three mussel species and their fish hosts, namely that they each use hosts with different dispersal capabilities, ranging from low, moderate, and high, respectively. The low divergence of mtDNA sequence variation reported in this and other recent mussel studies indicates that considerable extant population genetic variation probably originated during the late Pleistocene and Holocene. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 114 , 376–397.     

Genetic characterization and population bottleneck in the Hucul horse based on microsatellite and mitochondrial data

The aim of this work was to gather information about the origin and genetic characterization of the Central European Hucul horse based on 71 horses using 17 microsatellites and the D‐loop region of mtDNA. Their genetic relationship to the Polish Konik (N = 7), German (N = 4) and Hungarian wild Przewalski horses (N = 4) and 200 horse sequences from GenBank was also analysed. Both microsatellite and mtDNA analysis showed a high genetic variation in the Hucul. A total of 130 alleles were detected, the mean number of observed alleles per microsatellite was 7.647, and the number of effective alleles was 4.401. The average observed and expected heterozygosity were 0.706 and 0.747, respectively. The high heterozygosity values and Wright's fixation index (F_IS) (?0.128) indicated a low level of inbreeding, low or no selection pressure, and large number of alleles. mtDNA analysis revealed 18 haplotypes for the Hucul population with a total of 23 variable sites. Haplotype and nucleotide diversities were 0.935 ± 0.011 and 0.022 ± 0.012, respectively. Neutrality tests (Tajima's D and Fu's Fs) were non‐significant, and mismatch distribution was ragged, indicating that the Hucul population is in genetic equilibrium. The most frequent mtDNA D‐loop region belonged to haplogroup A (48%), which was also present in Przewalski Wild horse samples, while Polish Konik samples belonged to three haplotypes and C, F, and G haplogroups. Large and significant pairwise Φ_ST values along with a small number of common haplotypes indicated a low level of gene flow and lack of genetic structure among the three studied breeds (Hucul, Konik, and Przewalski Wild horse). The present work contributes to our knowledge of the genetic diversity of the Hucul horse and helps to define its genetic conservation. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 54–65.     

Systematics and phylogeography of the Taiwanese endemic minnow Candidia barbatus (Pisces: Cyprinidae) based on DNA sequence,allozymic, and morphological analyses

The phylogeographical patterns of the endemic minnow, Candidia barbatus, were examined using mitochondrial (mt) cytochrome b sequences, allozyme electrophoresis, and morphology. The 23 mtDNA haplotypes recovered were grouped into six lineages (A–F). According to a molecular clock of 1.7% per million years, we estimated that the colonization of C. barbatus to Taiwan took place three million years ago with several subsequent vicariance events. In the allozyme analysis, the Bayesian approach partitioned different populations into three clusters, with clusters I, II, and III mainly found in northern, central, and southern Taiwan, respectively. Morphologically, fish from north and south of the Tropic of Cancer had distinct numbers of lateral line scales and were separated into two morphs. The cohabitation of different mtDNA lineages with high allozymic and morphological similarities suggests that interbreeding amongst them is possible. Therefore, in spite of highly differentiated mtDNA, samples from different collecting sites should be considered a single species. Based on the current population distribution and levels of divergence, we propose that both orogenic activities and marine regression during glacial epoch were the two main forces shaping population differentiation of this species. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 161 , 613–632.     

The genus Huperzia (Lycopodiaceae) in the Azores and Madeira

The taxonomy and nomenclature of the genus Huperzia Bernh. in the Azores and Madeira have been reviewed. Plants collected in the Azores and Madeira were characterized morphologically. The independence between two endemic species common to Madeira and the Azores Islands –Huperzia suberecta (Lowe) Tardieu and Huperzia dentata (Herter) Holub – is clearly shown. A clear‐cut morphological separation between these taxa and Huperzia selago (L.) Bernh. ex Schrank & Mart. of continental Europe is established. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 522–533.     

Field observation of foraging behavior by a group of adult diving beetles Agabus (Gaurodytes) bipustulatus preying on an adult Lissotriton boscai

Adult dytiscids prey on a variety of items including other invertebrates but also larger prey such as frogs and fish. Observations of anuran larvae predation are common. However, there have been no reports concerning predation on caudata post‐metamorphosis by dytiscids. Here we describe a predation event by a group of adult diving beetles of the species Agabus (Gaurodytes) bipustulatus (Linnaeus, 1767) on an adult Lissotriton boscai (Lataste in Tourneville, 1879). This report represents the first observation of foraging behavior of adult diving beetles preying on and consuming a living post‐metamorphic newt.     

A mark and recapture study of water beetles (Coleoptera: Dytiscidae) in a group of semi-permanent and temporary ponds

A year mark and recapture study of aquatic Dytiscidae (Coleoptera) in a group of seven semi-permanent and temporary ponds in Cheshire (UK) is described. Pond water levels showed strong seasonal fluctuations with most ponds drying and refilling on several occasions over the study period. Sampling and labelling of aquatic Dytiscidae was carried out at monthly intervals. Beetles were labelled by fixing numbered waterproof paper labels to the elytra with cyanoacrylate `Superglue'. A total of 780 beetles were marked and released of which 11.5% were subsequently recaptured over varying time intervals of one month to eighteen months. The six species marked (Agabus bipustulatus (Linnaeus, 1767), Acilius sulcatus (Linnaeus, 1758), Colymbetes fuscus (Linnaeus, 1758), Dytiscus marginalis (Linnaeus, 1758), Ilybius ater (De Geer, 1774) and Ilybius fuliginosus (Fabricius, 1792)) are all known to be highly flight mobile. Colymbetes fuscus did not move between ponds as a result of ponds drying up in the summer. The behaviour of Dytiscus marginalis and Acilius sulcatus was found to be in broad agreement with previous accounts of dispersal from over-wintering ponds into temporary ponds in the spring with movement back to permanent ponds in the summer as the temporary ponds dry up. In contrast, Agabus bipustulatus did not migrate when ponds dried up in the summer but remained in terrestrial vegetation in damp pond basins for several months after open water had disappeared, only migrating if the basin dried out completely. Both species of Ilybius were captured and marked in very low numbers and no recaptures were made. As more ponds became temporary over the years of the study Agabus bipustulatus persisted while the other five species declined or were lost altogether.     

Predation by introduced fish constrains the thermal distribution of aquatic Coleoptera in mountain lakes

1. Most high mountain lakes were free of fish until humans stocked them. This provides the opportunity to study the extent to which predation constrains the thermal distribution of large macroinvertebrates, among lakes of a suitable temperature for them. 2. We analysed the distribution of aquatic beetles (Coleoptera, Dytisicidae, including the genera Agabus, Platambus, Hydroporus and Boreonectes) in a set of 82 lakes in the Pyrenees. Temperature was the most important variable explaining the overall distribution of dytiscids (out of 29 environmental variables considered). The presence of fish (i.e. salmonids) and macrophyte cover (plants provide refuge from predators) was also important. Indeed, either salmonids or macrophytes as single factors explained more of the variance in dytiscid distribution than temperature. 3. Further analysis showed that the presence of salmonids was particularly important for the medium‐sized dytiscid Agabus bipustulatus. This species is generally eurythermic, although in the Pyrenees it is found mainly in cold lakes. This is attributed to predation pressure from fish, since the latter are more likely to be present in warm lakes. As a consequence, salmonids increase fragmentation of A. bipustulatus with respect to populations occupying lowland habitats. 4. Predation can therefore constrain the distribution of species to a fraction of habitats that appear suitable based on simple thermal responses. As humans also facilitate the dispersal of non‐native fish, the consequent modification of distribution patterns in fresh waters could make it difficult to predict how distributions might change in relation to climate warming.     

Phylogeography of the brown hare (Lepus europaeus) in Europe: a legacy of south‐eastern Mediterranean refugia?

Aim We analysed the population genetics of the brown hare (Lepus europaeus) in order to test the hypothesis that this species migrated into central Europe from a number of late glacial refugia, including some in Asia Minor. Location Thirty‐three localities in Greece, Bulgaria, Italy, Croatia, Serbia, Poland, Switzerland, Austria, France, Germany, the Netherlands, Spain, the United Kingdom, Turkey and Israel. Methods In total, 926 brown hares were analysed for mitochondrial DNA (mtDNA) variation by restriction fragment length polymorphism (RFLP) performed on polymerase chain reaction‐amplified products spanning cytochrome b (cyt b)/control region (CR), cytochrome oxidase I (COI) and 12S–16S rRNA. In addition, sequence analysis of the mtDNA CR‐I region was performed on 69 individuals, and the data were compared with 137 mtDNA CR‐I sequences retrieved from GenBank. Results The 112 haplotypes detected were partitioned into five phylogeographically well‐defined major haplogroups, namely the ‘south‐eastern European type haplogroup’ (SEEh), ‘Anatolian/Middle Eastern type haplogroup’ (AMh), ‘European type haplogroup, subgroup A’ (EUh‐A), ‘European type haplogroup, subgroup B’ (EUh‐B) and ‘Intermediate haplogroup’ (INTERh). Sequence data retrieved from GenBank were consistent with the haplogroups determined in this study. In Bulgaria and north‐eastern Greece numerous haplotypes of all five haplogroups were present, forming a large overlap zone. Main conclusions The mtDNA results allow us to infer post‐glacial colonization of large parts of Europe from a late glacial/early Holocene source population in the central or south‐central Balkans. The presence of Anatolian/Middle Eastern haplotypes in the large overlap zone in Bulgaria and north‐eastern Greece reveals gene flow from Anatolia to Europe across the late Pleistocene Bosporus land‐bridge. Although various restocking operations could be partly responsible for the presence of unexpected haplotypes in certain areas, we nevertheless trace a strong phylogeographic signal throughout all regions under study. Throughout Europe, mtDNA results indicate that brown hares are not separated into discernable phyletic groups.     

Of mice and the ‘Age of Discovery’: the complex history of colonization of the Azorean archipelago by the house mouse (Mus musculus) as revealed by mitochondrial DNA variation

Humans have introduced many species onto remote oceanic islands. The house mouse (Mus musculus) is a human commensal and has consequently been transported to oceanic islands around the globe as an accidental stowaway. The history of these introductions can tell us not only about the mice themselves but also about the people that transported them. Following a phylogeographic approach, we used mitochondrial D‐loop sequence variation (within an 849‐ to 864‐bp fragment) to study house mouse colonization of the Azores. A total of 239 sequences were obtained from all nine islands, and interpretation was helped by previously published Iberian sequences and 66 newly generated Spanish sequences. A Bayesian analysis revealed presence in the Azores of most of the D‐loop clades previously described in the domesticus subspecies of the house mouse, suggesting a complex colonization history of the archipelago as a whole from multiple geographical origins, but much less heterogeneity (often single colonization?) within islands. The expected historical link with mainland Portugal was reflected in the pattern of D‐loop variation of some of the islands but not all. A more unexpected association with a distant North European source area was also detected in three islands, possibly reflecting human contact with the Azores prior to the 15th century discovery by Portuguese mariners. Widening the scope to colonization of the Macaronesian islands as a whole, human linkages between the Azores, Madeira, the Canaries, Portugal and Spain were revealed through the sharing of mouse sequences between these areas. From these and other data, we suggest mouse studies may help resolve historical uncertainties relating to the ‘Age of Discovery’.     

Genetic structure of house mouse (Mus musculus Linnaeus 1758) populations in the Atlantic archipelago of the Azores: colonization and dispersal

We analyzed the genetic structure and relationships of house mouse (Mus musculus) populations in the remote Atlantic archipelago of the Azores using nuclear sequences and microsatellites. We typed Btk and Zfy2 to confirm that the subspecies Mus musculus domesticus was the predominant genome in the archipelago. Nineteen microsatellite loci (one per autosome) were typed in a total of 380 individuals from all nine Azorean islands, the neighbouring Madeiran archipelago (Madeira and Porto Santo islands), and mainland Portugal. Levels of heterozygosity were high on the islands, arguing against population bottlenecking. The Azorean house mouse populations were differentiated from the Portuguese and Madeiran populations and no evidence of recent migration between the three was obtained. Within the Azores, the Eastern, Western, and Central island groups tended to act as separate genetic units for house mice, with some exceptions. In particular, there was evidence of recent migration events among islands of the Central island group, whose populations were relatively undifferentiated. Santa Maria had genetically distinctive mice, which may relate to its colonization history. © 2013 The Linnean Society of London     

No genetic differentiation between geographically isolated populations of Clarias macrocephalus Günther in Malaysia revealed by sequences of mtDNA Cytochrome b and D‐loop gene regions

In the present study, we assessed the genetic variation of three Clarias macrocephalus Günther populations collected from Kedah, Perlis and Kelantan (Peninsular Malaysia) using sequences of partial mitochondrial cytochrome b (Cyt b) and D‐loop genes. A total of 57 individuals were sequenced and 1470 bp were obtained (1053 bp Cyt‐b; 417 bp D‐loop). The analysis revealed 21 haplotypes based on 81 polymorphic sites. Nucleotide diversity (π) was 0.003 in all populations while haplotype diversity ranged from 0.657 to 0.765. No significant genetic differentiation among the three populations was observed. Nevertheless, a number of private haplotypes was discovered, providing valuable information for selective breeding programs.     

Multilocus phylogeography and population structure of common eiders breeding in North America and Scandinavia

Aim Glacial refugia during the Pleistocene had major impacts on the levels and spatial apportionment of genetic diversity of species in northern latitude ecosystems. We characterized patterns of population subdivision, and tested hypotheses associated with locations of potential Pleistocene refugia and the relative contribution of these refugia to the post‐glacial colonization of North America and Scandinavia by common eiders (Somateria mollissima). Specifically, we evaluated localities hypothesized as ice‐free areas or glacial refugia for other Arctic vertebrates, including Beringia, the High Arctic Canadian Archipelago, Newfoundland Bank, Spitsbergen Bank and north‐west Norway. Location Alaska, Canada, Norway and Sweden. Methods Molecular data from 12 microsatellite loci, the mitochondrial DNA (mtDNA) control region, and two nuclear introns were collected and analysed for 15 populations of common eiders (n = 716) breeding throughout North America and Scandinavia. Population genetic structure, historical population fluctuations and gene flow were inferred using F‐statistics, analyses of molecular variance, and multilocus coalescent analyses. Results Significant inter‐population variation in allelic and haplotypic frequencies were observed (nuclear DNA F_ST = 0.004–0.290; mtDNA Φ_ST = 0.051–0.927). Whereas spatial differentiation in nuclear genes was concordant with subspecific designations, geographic proximity was more predictive of inter‐population variance in mitochondrial DNA haplotype frequency. Inferences of historical population demography were consistent with restriction of common eiders to four geographic areas during the Last Glacial Maximum: Belcher Islands, Newfoundland Bank, northern Alaska and Svalbard. Three of these areas coincide with previously identified glacial refugia: Newfoundland Bank, Beringia and Spitsbergen Bank. Gene‐flow and clustering analyses indicated that the Beringian refugium contributed little to common eider post‐glacial colonization of North America, whereas Canadian, Scandinavian and southern Alaskan post‐glacial colonization is likely to have occurred in a stepwise fashion from the same glacial refugium. Main conclusions Concordance of proposed glacial refugia used by common eiders and other Arctic species indicates that Arctic and subarctic refugia were important reservoirs of genetic diversity during the Pleistocene. Furthermore, suture zones identified at MacKenzie River, western Alaska/Aleutians and Scandinavia coincide with those identified for other Arctic vertebrates, suggesting that these regions were strong geographic barriers limiting dispersal from Pleistocene refugia.     

Deep intra‐island divergence of a montane forest endemic: phylogeography of the Puerto Rican frog Eleutherodactylus portoricensis (Anura: Eleutherodactylidae)

Aim Hypotheses proposed for lineage diversification of tropical montane species have rarely been tested within oceanic islands. Our goal was to understand how basin barriers and Pleistocene climatic fluctuations shaped the distribution of diversity in Eleutherodactylus portoricensis (Eleutherodactylidae), a frog endemic to the montane rain forests of Puerto Rico. Location The north‐eastern (Luquillo) and south‐eastern (Cayey) mountains of Puerto Rico. Methods We generated mitochondrial DNA (mtDNA) control region sequences (c. 565 bp) from 144 individuals of E. portoricensis representing 16 localities, and sequenced 646 bp of cytochrome b and 596 bp of nuclear DNA (nDNA) rhodopsin exon and intron 1 from a subset of individuals. We conducted a phylogenetic analysis on the mtDNA sequence data and explored population substructure with maximum parsimony networks, a spatial analysis of molecular variance, and pairwise F_ST analysis. Coalescent simulations were performed to test alternative models of population divergence in response to late Pleistocene interglacial periods. Historical demography was assessed through coalescent analyses and Bayesian skyline plots. Results We found: (1) two highly divergent groups associated with the disjunct Luquillo and Cayey Mountains, respectively; (2) a shallow mtDNA genetic discontinuity across the La Plata Basin within the Cayey Mountains; (3) phylogeographic congruence between nDNA and mtDNA markers; (4) divergence dates for both mtDNA and nDNA pre‐dating the Holocene interglacial (c. 10 ka), and nDNA suggesting divergence in the penultimate interglacial (c. 245 ka); and (5) historical demographic stability in both lineages. Main conclusions The low‐elevation Caguas Basin is a long‐term barrier to gene flow between the two montane frog populations. Measures of genetic diversity for mtDNA were similar in both lineages, but lower nDNA diversity in the Luquillo Mountains lineage suggests infrequent dispersal between the two mountain ranges and colonization by a low‐diversity founder population. Population divergence began prior to the Holocene interglacial. Stable population sizes over time indicate a lack of demonstrable demographic response to climatic changes during the last glacial period. This study highlights the importance of topographic complexity in promoting within‐island vicariant speciation in the Greater Antilles, and indicates long‐term persistence and lineage diversification despite late Pleistocene climatic oscillations.     

The divergence of two independent lineages of an endemic Chinese gecko,Gekko swinhonis,launched by the Qinling orogenic belt

The genetic structure and demographic history of an endemic Chinese gecko, Gekko swinhonis, were investigated by analysing the mitochondrial cytochrome b gene and 10 microsatellite loci for samples collected from 27 localities. Mitochondrial DNA data provided a detailed distribution of two highly divergent evolutionary lineages, between which the average pairwise distance achieved was 0.14. The geographic division of the two lineages coincided with a plate boundary consisting of the Qinling and Taihang Mts, suggesting a historical vicariant pattern. The orogeny of the Qinling Mts, a dispersal and major climatic barrier of the region, may have launched the independent lineage divergence. Both lineages have experienced recent expansion, and the current sympatric localities comprised the region of contact between the lineages. Individual‐based phylogenetic analyses of nucDNA and Bayesian‐clustering approaches revealed a deep genetic structure analogous to mtDNA. Incongruence between nucDNA and mtDNA at the individual level at localities outside of the contact region can be explained by the different inheritance patterns and male‐biased dispersal in this species. High genetic divergence, long‐term isolation and ecological adaptation, as well as the morphological differences, suggest the presence of a cryptic species.     

Two synonymy cases within the genus Neoseiulella (Acari: Phytoseiidae): is the molecular evidence so evident?

Synonymy is considered as a great problem for diversity characterization and further applications, especially for biological control success with regard to the accurate identification of natural enemies. The present study focuses on two synonymy cases of natural enemies, belonging to the family Phytoseiidae, the genus Neoseiulella, specifically five species of this genus: Neoseiulella tiliarum, Neoseiulella formosa, Neoseiulella aceri, Neoseiulella squamiger, and Neoseiulella aceris. Morphological and molecular analyses [12S rRNA, cytochrome b (Cytb) mitochondrial (mt)DNA, internal transcribed spacer DNA] were applied. First, the results obtained support the synonymy of N. tiliarum and N. formosa. Second, because morphological differences (solenostomes occurrence) were observed for the first time between the type material of N. aceris and N. squamiger, the present study does not provide sufficient evidence to synomymize these two species and further analyses are required. Lastly, we assume that N. squamiger and N. aceri are synonyms. However, three groups of specimens, including N. aceri and N. squamiger, were identified by two mitochondrial DNA genes (12S rRNA and Cytb mtDNA). Therefore, the present study highlights the problems encountered when using only mitochondrial genes to diagnose species. The great mtDNA variations observed appear to reflect population differentiation (linked to plant support). This is the first time that such high intraspecific differentiation is be observed within the family Phytoseiidae. Further experiments, such as cross‐breeding and microsatellite DNA marker analyses, are planned to characterize gene flow and reproductive isolation levels within this species. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 323–344.     

A new cytochrome b phylogroup of the common vole (Microtus arvalis) endemic to the Balkans and its implications for the evolutionary history of the species

The phylogeographic architecture of the common vole, Microtus arvalis, has been well‐studied using mitochondrial DNA and used to test hypotheses relating to glacial refugia. The distribution of the five described cytochrome b (cyt b) lineages in Europe west of Russia has been interpreted as a consequence of postglacial expansion from both southern and central European refugia. A recently proposed competing model suggests that the ‘cradle’ of the M. arvalis lineages is in western central Europe from where they dispersed in different directions after the Last Glacial Maximum. In the present study, we report a new cyt b lineage of the common vole from the Balkans that is not closely related to any other lineage and whose presence might help resolve these issues of glacial refugia. The Balkan phylogroup occurs along the southern distributional border of M. arvalis in central and eastern Bosnia and Herzegovina, Montenegro, and eastern Serbia. Further north and west in Slovenia, Bosnia and Herzegovina, and Serbia, common voles belong to the previously‐described Eastern lineage, whereas both lineages are sympatric in one site in Bosnia and Herzegovina. The Balkan phylogroup most reasonably occupied a glacial refugium already known for various Balkan endemic species, in contrast to the recently proposed model. South‐east Europe is an absolutely crucial area for understanding the postglacial colonization history of small mammals in Europe and the present study adds to the very few previous detailed phylogeographic studies of this region. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 788–796.     

Multiple ice‐age refugia in Pacific cod,Gadus macrocephalus

Pleistocene ice‐ages greatly influenced the historical abundances of Pacific cod, Gadus macrocephalus, in the North Pacific and its marginal seas. We surveyed genetic variation at 11 microsatellite loci and mitochondrial (mt) DNA in samples from twelve locations from the Sea of Japan to Washington State. Both microsatellite (mean H = 0.868) and mtDNA haplotype (mean h = 0.958) diversities were large and did not show any geographical trends. Genetic differentiation between samples was significantly correlated with geographical distance between samples for both microsatellites (F_ST = 0.028, r² = 0.33) and mtDNA (F_ST = 0.027, r² = 0.18). Both marker classes showed a strong genetic discontinuity between northwestern and northeastern Pacific populations that likely represents groups previously isolated during glaciations that are now in secondary contact. Significant differences appeared between samples from the Sea of Japan and Okhotsk Sea that may reflect ice‐age isolations in the northwest Pacific. In the northeast Pacific, a microsatellite and mtDNA partition was detected between coastal and Georgia Basin populations. The presence of two major coastal mtDNA lineages on either side of the Pacific Ocean basin implies at least two ice‐age refugia and separate postglacial population expansions facilitated by different glacial histories. Northward expansions into the Gulf of Alaska were possible 14–15 kyr ago, but deglaciation and colonization of the Georgia Basin probably occurred somewhat later. Population expansions were evident in mtDNA mismatch distributions and in Bayesian skyline plots of the three major lineages, but the start of expansions appeared to pre‐date the last glacial maximum.