A phylogenetic study of the genus Apodemus by sequencing the mitochondrial DNA control region

The sequences of the mitochondrial DNA control region (D-loop) and flanking tRNA genes (about 1000 bp) of 20 samples of wood mice (genus Apodemus ) were analyzed in order to clarify the relationships between different species belonging to the genus. The phylogenetic trees obtained using different methods showed similar topologies with distinct Karstomys ( Apodemus epimelas and Apodemus mystacinus ) and Sylvaemus ( Apodemus alpicola , Apodemus flavicollis , Apodemus hermonensis , Apodemus sylvaticus and Apodemus uralensis ) subtrees. Within Sylvaemus all species appeared to be closely related to each other, probably as result of a bush-like radiation event. Nevertheless, A. hermonensis seemed to be the first diverging branch followed by A. sylvaticus ; A. alpicola and A. flavicollis appeared to be very closely related. Three individuals of uncertain taxonomical status were included in the analysis: hypotheses as to their status are discussed. Further phylogenetic analysis was carried out combining the D-loop sequences of part of the samples of certain taxonomical status with 12S rRNA and cytochrome b sequences obtained by other researchers. Furthermore, I present a structure analysis of the D-loop in Apodemus as compared other rodent species.     

Genetic variation and evolution in the genus Apodemus (Muridae: Rodentia)

Genetic variation was studied using protein electrophoresis of 28–38 gene loci in 1347 specimens of Apodemus agrar-ius, A. peninsulae, A. flavicollis, A. sylvaticus, A. alpicola, A. uralensis, A . cf. hyrcanicus, A. hermonensis, A. m. mystacinus and A. m. epimelas , representing 121 populations from Europe, the Middle East, and North Africa. Mean values of heterozygosity per locus for each species ranged from 0.02 to 0.04. Mean values of Nel's genetic distance ( D ) between the taxa ranged from 0.06 (between A. flavicollis and A. alpicola) to 1.34 (between A. uralensis and A. agrarius ). The highest values of D were found between A. agrarius and other Apodemus species (0.62-1.34). These values correspond to those generally observed between genera in small mammals. Our data show that A. agrarius and A. peninsulae are sister species, well-differentiated from other taxa. High genetic distance between A. m. mystacinus and A. m. epimelas leads us to consider them distinct species and sister taxa to other Western Palaearctic species of the subgenus Sylvaemus . The data also suggest a recent separation of members of the latter group from a common ancestor, and subsequent rapid radiation, making it difficult to infer phylogenetic relationships. Some taxonomic implications of the results are discussed further.© 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 75 , 395–419.     

The use of arboreal runways by the woodland rodents, Apodemus sylvaticus (L.), A. flavicollis (Melchior) and Clethrionomys glareolus (Schreber)

The use of arboreal runways by the rodents, Apodemus sylvaticus, A. flavicollis and Clethrionomys glareolus , was investigated in the deciduous woodland of Woodchester Park, Gloucestershire, England. Longworth live traps were tied to horizontal and inclining branches of trees and shrubs up to 3 m. Arboreality was extensive in all three species, although less in C. glareolus than in either Apodemus species. There was no indication that A. flavicollis consistently used arboreal runways more than A. sylvaticus but the data suggest that the peak of arboreal activity of these congeners occurred in different seasons. In A. sylvaticus , males were more arboreal than females and adults were more arboreal than juveniles. In, 4. flavicollis , juveniles were trapped significantly more often on arboreal runways than adults. The reasons for, and the consequences of, arboreality in small rodents are discussed.     

Phylogeny of the genus Apodemus with a special emphasis on the subgenus Sylvaemus using the nuclear IRBP gene and two mitochondrial markers: cytochrome b and 12S rRNA

Phylogenetic relationships among 17 extant species of Murinae, with special reference to the genus Apodemus, were investigated using sequence data from the nuclear protein-coding gene IRBP (15 species) and the two mitochondrial genes cytochrome b and 12S rRNA (17 species). The analysis of the three genes does not resolve the relationships between Mus, Apodemus, and Rattus but separates Micromys from these three genera. The analysis of the two mitochondrial regions supported an association between Apodemus and Tokudaia and indicated that these two genera are more closely related to Mus than to Rattus or Micromys. Within Apodemus, the mitochondrial data sets indicated that 8 of the 9 species analyzed can be sorted into two main groups: an Apodemus group, with A. agrarius, semotus, and peninsulae, and a Sylvaemus group, with uralensis, flavicollis, alpicola, sylvaticus, and hermonensis. The position of Apodemus mystacinus is ambiguous and might be either included in Sylvaemus or considered a distinct subgenus, Karstomys, more closely related to Sylvaemus than to Apodemus. Estimation of the divergence time for these taxa suggests a separation between 7 and 8 My ago for the three groups (mystacinus and the two subgenera Apodemus and Sylvaemus). Within each subgenus, divergence times are between 5.4 and 6 My for Apodemus and between 2.2 and 3.5 My for Sylvaemus and mystacinus.     

Molecular phylogeny and distribution of three Apodemus species (Muridae, Rodentia) in Serbia

Phylogenetic relationships among three species of the genus Apodemus ( A. flavicollis, A. sylvaticus and A. agrarius ) from seven different localities in Serbia were estimated using arbitrarily primed PCR (AP-PCR) profiling, which allowed clear distinction between the sibling species A. flavicollis and A. sylvaticus . Eight oligonucleotide primers out of the 22 screened, gained a total of 397 amplified bands. The average pairwise genetic distances within Apodemus species ranged from 0.51 between A. flavicollis and A. sylvaticus , to 0.74 between A. sylvaticus and A. agrarius. The large genetic distances obtained among populations of A. flavicollis (0.13) indicate that this part of the Balkan Peninsula was probably a refuge during the last glaciation. Furthermore, differences obtained between populations south and north from the river Danube suggest that this species survived the Last Glacial Maximum in at least two separate refuges. Although our data for molecular markers are from a limited number of localities, extended analyses of C-band distribution on chromosomes from previously studied localities south of the rivers Danube and Sava, indicate that distribution of A. sylvaticus is limited to northern Serbia. Therefore, it is possible that this part of the Balkan Peninsula was not a refuge for this species during the last glaciation.     

Spatial organization in sympatric populations of Apodemus sylvaticus and A. flavicollis (Rodentia: Muridae)

The spatial organization and relationships of Apodemus sylvaticus and A. flavicollis in mixed deciduous woodland were examined using live-trapping techniques. Each population was aggregated and contagion was greatest during early summer. Aggregation in A sylvaticus varied inversely with density so that at high densities this species was randomly dispersed. In A. flavicollis aggregation increased with increasing density. This interspecific difference may result from differing annual population cycles. Adult males, adult females and juveniles had different patterns of dispersion and association in A. sylvaticus and A. flavicollis suggesting differences in their social behaviour. Intraspecific associations, particularly among males and between males and females, made a major contribution to overall contagion in the populations. A. sylvaticus and A. flavicollis were segregated throughout the year except for late summer and they were also prevalent in different cover types. The behaviour which determined the spatial segregation of the species may be acquired since juvenile A. sylvaticus were associated more frequently with A. flavicollis and juvenile A. flavicollis were associated less frequently with A. sylvaticus than their respective adult conspecifics. Comparing sympatric with allopatric populations of A. sylvaticus , and observing inter-specific behaviour, suggest that the spatial segregation in sympatry results from A. sylvaticus avoiding the competitively superior A. flavicollis .     

Molecular phylogeny and taxonomy of wood mice (genus Apodemus Kaup, 1829) based on complete mtDNA cytochrome b sequences, with emphasis on Chinese species

Phylogenetic relationships among 15 species of wood mice (genus Apodemus) were reconstructed to explore some long-standing taxonomic problems. The results provided support for the monophyly of the genus Apodemus, but could not reject the hypothesis of paraphyly for this genus. Our data divided the 15 species into four major groups: (1) the Sylvaemus group (A. sylvaticus, A. flavicollis, A. alpicola, and A. uralensis), (2) the Apodemus group (A. peninsulae, A. chevreri, A. agrarius, A. speciosus, A. draco, A. ilex, A. semotus, A. latronum, and A. mystacinus), (3) A. argenteus, and (4) A. gurkha. Our results also suggested that orestes should be a valid subspecies of A. draco rather than an independent species; in contrast, A. ilex from Yunnan may be regarded as a separate species rather than a synonym of orestes or draco. The species level status of A. latronum, tscherga as synonyms of A. uralensis, and A. chevrieri as a valid species and the closest sibling species of A. agrarius were further corroborated by our data. Applying a molecular clock with the divergences of Mus and Rattus set at 12 million years ago (Mya) as a calibration point, it was estimated that five old lineages (A. mystacinus and four major groups above) diverged in the late Miocene (7.82-12.74 Mya). Then the Apodemus group (excluding A. mystacinus) split into two subgroups: agrarius and draco, at about 7.17-9.95 Mya. Four species of the Sylvaemus group were estimated to diverge at about 2.92-5.21 Mya. The Hengduan Mountains Region was hypothesized to have played important roles in Apodemus evolutionary histories since the Pleistocene.     

Genetic relationships between European and Transcaucasian mice of the genus Apodemus Kaup]

Genetic relations between four European mice species of the genus Apodemus (Apodemus sylvaticus, A. flavicollis, A. microps and A. falzfeini) and five Transcaucasian ones (A. mystacinus, A. microps, and the forms No 2, 3p, 3f) were studied for 37 biochemical loci. Close genetic relations were demonstrated between the mice of subgenus Sylvaemus and A. mystacinus. A. microps and A. falzfeini from the Caucasus were shown conspecific to these species from the Ukraine and the presence of the two new different species--the form No 2 and 3p in the Caucasus was established.     

Population structure and dynamics of sympatric Apodemus species (Rodentia: Muridae)

This paper describes the population biology of sympatric populations of Apodemus sylvaticus and A. flavicollis in western England. Annual changes in population siie closely resembled those seen in allopatric populations such that simultaneous peaks in number occurred only in autumn and early winter. Numbers of A. sylvaticus were low and stable in early summer increasing rapidly in late summer and autumn and remaining high throughout winter before a sharp decline in spring. Numbers of A. flavicollis increased after the start of reproduction and continued to rise throughout summer and autumn. The period of major decline of this species was during early winter. Densities of A. sylvaticus and A. flavicollis were comparable with those cited in allopatric studies. Spring populations of both species consisted of cohorts first appearing in the late summer and autumn of the previous year. Few individuals present in spring survived the summer. Reproduction of A. flavicollis commenced four to six weeks before that of A. sylvaticus . Seasonal variation in mean weight of adult males and females also indicates that those changes, due to the acquisition of secondary sex characteristics and reproductive maturity, occur earlier in A. flavicollis than in A. sylvaticus . Data presented contain no evidence of interspecific competition and it is concluded that interspecific differences, chiefly asynchrony in reproduction and annual population cycles, contribute to their stable coexistence.     

The importance of Laelaps agilis C.L. Koch 1836 (Mesostigmata: Parasitiformae) as a vector of Hepatozoon sylvatici Coles 1914 (Sporozoa: Haemogregarinidae) (author's transl)]

The importance of the mite Laelaps agilis (Parasitiformae) for the transmission of the Hepatozoon species living in the blood of Apodemus sylvaticus and Apodemus flavicollis (Hepatozoon sylvatici) is examined. With this mite the experimental transfer of the parasite to bred hosts of the same species was successful five times; for once, the transmission to a non-specific host (Clethrionomys glareolus) was also possible. Besides, an eventual stimulating effect emanating from the gonads of female mites to the development of parasites can be considered. Differences in the morphology of schizonts in the bone marrow and in the liver of infected Apodemus flavicollis are especially pointed out.     

Conservation of sequences in related genomes of Apodemus: constraints on the maintenance of satellite DNA sequences.

Satellites from two related species of the Apodemus genus, A. sylvaticus and A. flavicollis, have been analysed with restriction enzymes Taq I, Alu I and Hind III. The restriction maps are closely conserved between species and show a novel feature of two differing internal periodicities within a 375 base pair repeating unit detected by two different restriction enzymes. This places constraints on the introduction of the observed restriction sites according to current models such as unequal crossing-over. The implications of such a conserved sequence and its presence in other species are discussed.     

Host specificity and genealogy of the louse Polyplax serrata on field mice, Apodemus species: a case of parasite duplication or colonisation?

The genealogy, population structure and population dynamics of the sucking louse Polyplax serrata were analysed across four host species of the genus Apodemus. An analysis of 126 sequences of cytochrome c oxidase subunit I using phylogenetic approaches and haplotype networking revealed a clear structure of European samples, forming three distinct and genetically distant clades with different host specificities. Although a clear connection was detected between the host and parasite genealogies/phylogenies, a uniform pattern of co-speciation was not found. For example, a dramatic shift in the degree of host specificity was demonstrated for two related louse lineages living in sympatry and sharing one of their host species. While one of the louse lineages frequently parasitised two different host taxa (Apodemus sylvaticus and Apodemus flavicollis), the other louse lineage was strictly specific to A. flavicollis. The estimate of divergence time between the two louse lineages indicates that they may have arisen due to parasite duplication on A. flavicollis.     

The mammalian model for population studies of B chromosomes: the wood mouse (Apodemus)

The presence of B chromosomes was reported in six species of the genus Apodemus (A. peninsulae, A. agrarius, A. sylvaticus, A. flavicollis, A. mystacinus, A. argenteus). High frequencies of Bs were recorded particularly in A. peninsulae and A. flavicollis. The origin of Bs in Apodemus seems to be rather ancient, and it is possible that the supernumerary elements, and/or a tendency for their appearance, were inherited from the common ancestor of the extant species. We have not found any correlated changes between frequencies of Bs and the level of protein polymorphism and/or heterozygosity assessed in electrophoretic studies. No measurable effect of Bs on overall genetic variability was thus revealed in studied populations. The pattern of evolutionary dynamics of Bs can be distinctly different between geographical populations, and both the parasitic and the heterotic models can be applied to explain the maintenance of Bs in different populations. Further studies are desirable to improve our understanding of the complicated evolutionary dynamics of Bs in the Apodemus species. An essential condition for success in this respect is much more detailed information on inheritance and the molecular structure of Bs.     

Presence-absence versus presence-only modelling methods for predicting bird habitat suitability

Habitat suitability models can be generated using methods requiring information on species presence or species presence and absence. Knowledge of the predictive performance of such methods becomes a critical issue to establish their optimal scope of application for mapping current species distributions under different constraints. Here, we use breeding bird atlas data in Catalonia as a working example and attempt to analyse the relative performance of two methods: the Ecological Niche factor Analysis (ENFA) using presence data only and Generalised Linear Models (GLM) using presence/absence data. Models were run on a set of forest species with similar habitat requirements, but with varying occurrence rates (prevalence) and niche positions (marginality). Our results support the idea that GLM predictions are more accurate than those obtained with ENFA. This was particularly true when species were using available habitats proportionally to their suitability, making absence data reliable and useful to enhance model calibration. Species marginality in niche space was also correlated to predictive accuracy, i.e. species with less restricted ecological requirements were modelled less accurately than species with more restricted requirements. This pattern was irrespective of the method employed. Models for wide‐ranging and tolerant species were more sensitive to absence data, suggesting that presence/absence methods may be particularly important for predicting distributions of this type of species. We conclude that modellers should consider that species ecological characteristics are critical in determining the accuracy of models and that it is difficult to predict generalist species distributions accurately and this is independent of the method used. Being based on distinct approaches regarding adjustment to data and data quality, habitat distribution modelling methods cover different application areas, making it difficult to identify one that should be universally applicable. Our results suggest however, that if absence data is available, methods using this information should be preferably used in most situations.     

Permanent Genetic Resources added to the Molecular Ecology Resources Database 1 February 2010-31 March 2010

This article documents the addition of 228 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Anser cygnoides, Apodemus flavicollis, Athene noctua, Cercis canadensis, Glis glis, Gubernatrix cristata, Haliotis tuberculata, Helianthus maximiliani, Laricobius nigrinus, Laricobius rubidus, Neoheligmonella granjoni, Nephrops norvegicus, Oenanthe javanica, Paramuricea clavata, Pyrrhura orcesi and Samanea saman. These loci were cross-tested on the following species: Apodemus sylvaticus, Laricobius laticollis and Laricobius osakensis (a proposed new species currently being described).     

Geography and host biogeography matter for understanding the phylogeography of a parasite

The co-evolution between hosts and parasites has long been recognized as a fundamental driver of macro-evolutionary patterns of diversification. The effect of co-differentiation on parasite diversification is, however, often confounded by underlying geographic patterns of host distribution. In order to disentangle the confounding effects of allopatric versus host speciation, the mitochondrial cytochrome b (cyt b) gene was sequenced in seventy individuals of the parasitic nematode genus Heligmosomoides sampled in the six Apodemus mice species common in the western Palearctic region. The nuclear internal transcribed spacers (ITS) 1 and 2 were also sequenced in fifteen parasites to confirm the mitochondrial data. All lineages differentiated according to a geographic pattern and independently from the sampled host species. This suggests that host speciation did not involve concurrent parasite speciation. However, the geographic distribution range of some parasite lineages mirrors that of A. sylvaticus lineages in SW Europe, and that of A. flavicollis lineages in the Balkans and in the Middle East. Thus, regional co-differentiation likely occurred between the parasite and the two sister Apodemus hosts in different parts of their distribution range. We suggest that differences in regional abundances of A. sylvaticus and A. flavicollis are responsible for generating this pattern of regional co-differentiation. This study highlights the importance of integrating both geography and biogeographic information from potential hosts to better understand their parasite phylogeography.     

So close and so different: comparative phylogeography of two small mammal species, the yellow-necked fieldmouse (Apodemus flavicollis) and the woodmouse (Apodemus sylvaticus) in the Western Palearctic region

In Europe, concordant geographical distribution among genetic lineages within different species is rare, which suggests distinct reactions to Quaternary ice ages. This study aims to determine whether such a discrepancy also affects a pair of sympatric species, which are morphologically and taxonomically closely related but which have slight differences in their ecological habits. The phylogeographic structures of two European rodents, the Yellow-necked fieldmouse (A. flavicollis) and the woodmouse (Apodemus sylvaticus) were, therefore, compared on the basis of mitochondrial DNA cytochrome b (mtDNA cyt b) sequences (965 base pairs) from 196 specimens collected from 59 European localities spread throughout the species distributions. The results indicate that the two species survived in different ways through the Quaternary glaciations. A. sylvaticus survived in the Iberian Peninsula from where it recolonized almost all Europe at the end of the last glaciation. Conversely, the refuge from which A. flavicollis recolonized Europe, including northern Spain, during the Holocene corresponds to the Italo-Balkan area, where A. sylvaticus suffered a serious genetic bottleneck. This study confirms that even closely related species may have highly different phylogeographic histories and shows the importance of ecological plasticity of the species for their survival through climate change. Finally, it suggests that phylogeographic distinctiveness may be a general feature of European species.     

Molecular detection of Bartonella species infecting rodents in Slovenia

Rodents, collected in three zoogeographical regions across Slovenia, were tested for the presence of bartonellae using direct PCR-based amplification of 16S/23S rRNA gene intergenic spacer region (ITS) fragments from splenic DNA extracts. Bartonella DNA was detected in four species of rodents, Apodemus flavicollis, Apodemus sylvaticus, Apodemus agrarius and Clethrionomys glareolus, in all three zoogeographic regions at an overall prevalence of 40.4%. The prevalence of infection varied significantly between rodent species and zoogeographical regions. Comparison of ITS sequences obtained from bartonellae revealed six sequence variants. Four of these matched the ITS sequences of the previously recognized species, Bartonella taylorii, Bartonella grahamii, Bartonella doshiae and Bartonella birtlesii, but one was new. The identity of the bartonellae from which the novel ITS sequences was obtained were further assessed by sequence analysis of cell division protein-encoding gene (ftsZ) fragments. This analysis demonstrated that the strain is most likely a representative of possible new species within the genus.     

Experimental adiaspiromycosis of the common vole (Microtus arvalis) and other small wild mammals after intraperitoneal inoculation

After intraperitoneal inoculation ofE. crescens to nine species of wild small mammals, six species of rodents (Apodemus flavicollis, A. sylvaticus, Clethrionomys glareolus, Microtus agrestis, M. arvalis, Mus musculus) developed generalized adiaspiromycosis. The course of experimental infection corresponded to the infections of laboratory animals provoked in the same manner of inoculation. The authors studied the affliction of individual organs, the dynamics of growth of adiaspores in the organs of the abdominal cavity and in the lungs and followed up morphological changes in the adiaspores. In fresh cover glass preparations, the presence of manifestations was demonstrated justifying considerations on the capacity of multiplication of the adiaspiromycosis agent in the host organism.