Phylogeography of the Japanese giant flying squirrel, Petaurista leucogenys (Rodentia: Sciuridae): implication of glacial refugia in an arboreal small mammal in the Japanese Islands

To test the association between temperate forest dynamics and glacial refugia for arboreal small mammals, we studied the phylogeography of the Japanese giant flying squirrel ( Petaurista leucogenys ) using complete mitochondrial cytochrome b gene sequences (1140 bp). This squirrel is endemic to three of Japan's main islands: Honshu, Shikoku, and Kyushu. We examined 58 specimens of P. leucogenys collected from 40 localities in Japan. Additionally, two individuals with unknown sampling localities were included in phylogenetic analyses. There were 54 haplotypes of P. leucogenys. We found five major phylogroups (Northern, Central, South-eastern, South-western, and Southern). These phylogroups may have originated from glacial refugia during the Late Pleistocene. After the last glaciation, the Northern phylogroup, widely distributed in eastern Japan, could have extensively expanded northward from its refugia. By contrast, in western Japan, population expansion was restricted to western Japan. All members of four phylogroups existed in western Japan during glaciations. The complicated phylogeographical pattern of P. leucogenys populations originating from western Japan may have resulted from the long history.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 47–60.     

Phylogenetic Relationships among Asian species of Petaurista (Rodentia, Sciuridae), Inferred from Mitochondrial Cytochrome b Gene Sequences

To elucidate the phylogenetic relationships among four species belonging to the genus Petaurista (P. alborufus castaneus, P. alborufus lena, P. leucogenys leucogenys, P. leucogenys nikkonis, P. petaurista melanotus, and P. philippensis grandis), we investigated the partial sequences (1,068 bp) of the mitochondrial cytochrome b gene for these giant flying squirrels. Phylogenetic trees (NJ, MP, and ML trees) constructed from cytochrome b sequences indicated that P. leucogenys was grouped independently with other species, and that P. philippensis was most closely related to P. petaurista with 99-100% bootstrap values. In addition, two subspecies of P. alborufus did not form a single clade: P. alborufus castaneus from China was most distantly related to the other species, whereas P. alborufus lena from Taiwan was closely related to P. petaurista and P. philippensis with 82-90% bootstrap values. This result suggests that it is reasonable to regard P. alborufus lena as a distinct species from P. alborufus castaneus.     

Hepatocystis in Formosan Mammals with a Description of a New Species*

SYNOPSIS. Hepatocystis has been found extremely common in a number of species of Formosan mammals, notably the large leafnosed bat (Hipposideros armiger terasensis), the Formosan macaque (Macaca cyclopis), and various subspecies of the red-bellied tree squirrel (Callosciurus erythraeus). Less often infected were the long-winged bat (Miniopterus schreibersii) and the Formosan giant flying squirrel (Petaurista grandis). The species occurring in H. a. terasensis is believed to be new, and is named Hepatocystis hipposideri. It causes considerable enlargement (but not other changes) of the host cell, is often amoeboid, and usually has numerous fine pigment granules.     

Phylogeography of the Russian flying squirrel (Pteromys volans): implication of refugia theory in arboreal small mammal of Eurasia

A phylogeographical study of the Russian (Siberian) flying squirrel (Pteromys volans) was carried out using the complete mitochondrial (mt) cytochrome b gene sequences with special reference to the refugia theory for the arboreal traits of this species. We examined 31 specimens from 24 localities, resulting in 28 haplotypes. One breeding specimen with a unique haplotype from Hokkaido, Japan was included in the phylogenetic analysis. There were three mtDNA lineages: Hokkaido, Far Eastern, and northern Eurasia. Divergence data among lineages demonstrated that the Hokkaido group separated from the other groups during the Holsteinian interglacial. The phylogeographical pattern of P. volans is different from that previously reported for terrestrial rodents associated with treeless habitats. Unlike grasslands, forests decreased during glaciation and moved southward because of the cold and arid environmental conditions. The glacial refugia of P. volans would have been associated with forest dynamics in the Pleistocene.     

Evolution of the Cyprinella lutrensis species group. III. Geographic variation in the mitochondrial DNA of Cyprinella lutrensis— the influence of Pleistocene glaciation on population dispersal and divergence

We employed restriction site variation in mitochondrial (mt)DNA to determine if significant phylogeographic structure occurs in the North American cyprinid fish Cyprinella lutrensis. Digestion patterns from 16 restriction endonucleases identified fifty mtDNA haplotypes among 127 individuals of Cyprinella lutrensis assayed from localities in the Gulf Coastal Plain, the Great Plains, and the Central Lowlands. Nucleotide sequence divergence among haplotypes was highly variable (mean ± SE: 2.87%± 0.08; range: 0.14–9.24%). Maximum-parsimony analysis and the neighbour joining method of tree construction revealed three major groupings (clades) of haplotypes that differed in geographic distribution. Divergence estimates between the basal clade, comprised of haplotypes primarily from the Brazos River in east Texas, and the remaining two clades, place C. lutrensis in the western Gulf Coastal Plain prior to Pleistocene glaciation. Nucleotide sequence divergence between the second clade, comprised of haplotypes from the Trinity and Calcasieu rivers in east Texas and southwestern Louisiana, respectively, and the third clade (comprised primarily of haplotypes from localities north of Texas and affected directly by Pleistocene glaciation), suggest that C. lutrensis colonized gladated regions to the north during the mid- to late Pleistocene. This hypothesis is supported by levels of intrapopulational nucleotide diversity in geographic localities outside of Texas and by geological evidence. Despite marked geographic variation in morphometries, meristics, and nuptial coloration, mtDNA variation in glaciated regions was not geographically structured, and subspecies of C. lutrensis were not identifiable by phylogenetic analysis of mtDNA.     

韩国与中国东北地区松鼠线粒体DNA控制区的序列变异

为了研究松鼠东北亚种(Sciurus vulgaris manchuricusThomas)不同种群的序列变异水平并进一步确定分类地位,我们分析了韩国5个地点和中国东北2个地点的松鼠标本的线粒体DNA控制区的全序列(1 058 bp)。39个韩国松鼠标本显示出21种单倍型,这些单倍型间的平均Tamura-Nei距离为1·0%; 24个中国松鼠标本显示21种单倍型,单倍型间的平均Tamura-Nei距离为1·4% (1 058 bp的全序列中发生变异的位点有119个,占11·2%)。韩国松鼠和中国松鼠间的平均距离为1·3%。并且韩国和中国松鼠的所有42个单倍型形成了一个单系分支,Fst值为0·04,表明在两个国家的松鼠间没有发生遗传分化。因此,序列分析的分子生物学的结果支持现行的分类,即来自韩国的朝鲜亚种(S·v·coreae)是中国北部地区松鼠东北亚种(S·v·manchuricus)的同物异名。这还需要进一步对北朝鲜和中国东北其它地区更多标本的分子和形态学分析来验证这一结论。     

Italian Peninsula preserves an evolutionary lineage of the fat dormouse Glis glis L. (Rodentia: Gliridae)

The present study examines the population genetic structure of fifty‐nine specimens of Glis glis (Linneaus, 1766) from thirteen localities in central Europe, sequencing a 400‐bp segment of the mitochondrial cytochrome b (cyt b) gene and a 673‐bp segment of the cytochrome c oxidase subunit I (COI) gene. The consensus tree obtained from Bayesian analysis revealed a robust dichotomy, showing two sister groups: one clade includes samples from a wide geographical area, extending from north‐central Europe to northern Italy (major branch sensu Bilton), and the other comprises samples collected in central and southern Italy and in Sicily (Italian branch). According to the Tajima–Nei model, the two phylogroups were separated by a sequence divergence of 0.8% (cyt b) – 2.6% (COI), showing the COI gene to be more informative than cyt b. On a smaller geographical scale, the Italian clade was further substructured, displaying geographical differentiation along the Peninsula. The gene pool in this area was patchy; whereas populations from Sicily Island demonstrated fixed cyt b and COI haplotypes, assuming processes of isolation and selection. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 11–21.     

New Species of Eimeria (Protozoa: Eimeriidae) from Malaysian Squirrels*

SYNOPSIS. In a survey of 520 squirrels of 19 species from Malaysia, 6 new species of Eimeria were found. Three are described from the subfamily Petauristinae: E. kylopetis n. sp. from the red-cheeked flying squirrel Hylopetes spadiceus; E. aeromysis n. sp. from the large black flying squirrel Aeromys tephromelas; E. malayensis n. sp. from the spotted giant flying squirrel Petaurista elegans. Three are described from the subfamily Sciurinae: E. hippuri n. sp. from the horse-tailed squirrel Sundasciurus hippurus; and E. notati and E. pahangi n. spp. from the plantain squirrel Callosciurus notatus.     

Phylogeographic study of Chinese seabuckthorn (Hippophae rhamnoides subsp. sinensis Rousi) reveals two distinct haplotype groups and multiple microrefugia on the Qinghai‐Tibet Plateau

Historical climate change can shape the genetic pattern of a species. Studies on this phenomenon provide great advantage in predicting the response of species to current and future global climate change. Chinese seabuckthorn (Hippophae rhamnoides subsp. sinensis) is one of the most important cultivated plants in Northwest China. However, the subspecies history and the potential genetic resources within the subspecies range remain unclear. In this study, we utilized two intergenic chloroplast regions to characterize the spatial genetic distribution of the species. We found 19 haplotypes in total, 12 of which were unique to the Chinese seabuckthorn. The populations observed on the Qinghai‐Tibet Plateau (QTP) consisted of most of the haplotypes, while in the northeast of the range of the subspecies, an area not on the QTP, only four haplotypes were detected. Our study also revealed two distinct haplotype groups of the subspecies with a sharp transition region located in the south of the Zoige Basin. 89.96% of the genetic variation located between the regions. Mismatch analysis indicated old expansions of these two haplotype groups, approximately around the early stage of Pleistocene. Additional morphological proofs from existing studies and habitat differentiation supported a long independent colonization history among the two regions. Potential adaptation probably occurred but needs more genome and morphology data in future. Chinese seabuckthorn have an older population expansion compared with subspecies in Europe. The lack of large land ice sheets and the heterogeneous landscape of the QTP could have provided extensive microrefugia for Chinese seabuckthorn during the glaciation period. Multiple localities sustaining high‐frequency private haplotypes support this hypothesis. Our study gives clear insight into the distribution of genetic resources and the evolutionary history of Chinese seabuckthorn.     

Metagenomic analysis reveals a functional signature for biomass degradation by cecal microbiota in the leaf-eating flying squirrel (Petaurista alborufus lena)

ABSTRACT: BACKGROUND: Animals co-evolve with their gut microbiota; the latter can perform complex metabolic reactions that cannot be done independently by the host. Although the importance of gut microbiota has been well demonstrated, there is a paucity of research regarding its role in foliage-foraging mammals with a specialized digestive system. RESULTS: In this study, a 16S rRNA gene survey and metagenomic sequencing were used to characterize genetic diversity and functional capability of cecal microbiota of the folivorous flying squirrel (Petaurista alborufus lena). Phylogenetic compositions of the cecal microbiota derived from 3 flying squirrels were dominated by Firmicutes. Based on end-sequences of fosmid clones from 1 flying squirrel, we inferred that microbial metabolism greatly contributed to intestinal functions, including degradation of carbohydrates, metabolism of proteins, and synthesis of vitamins. Moreover, 33 polysaccharide-degrading enzymes and 2 large genomic fragments containing a series of carbohydrate-associated genes were identified. CONCLUSIONS: Cecal microbiota of the leaf-eating flying squirrel have great metabolic potential for converting diverse plant materials into absorbable nutrients. The present study should serve as the basis for future investigations, using metagenomic approaches to elucidate the intricate mechanisms and interactions between host and gut microbiota of the flying squirrel digestive system, as well as other mammals with similar adaptations.     

Multiple glacial refugia and postglacial colonization routes inferred for a woodland geophyte,Cyclamen purpurascens: patterns concordant with the Pleistocene history of broadleaved and coniferous tree species

The phylogeographical history of mid‐altitude woodland herbs that depend on moist and shaded forest habitats is poorly understood. Here, we analysed the genetic structure of Cyclamen purpurascens, a mountainous calcicolous perennial, to test hypotheses regarding its glacial survival in single or multiple refugia and postglacial colonization routes, and to explore how they are congruent with the histories inferred for temperate trees and other mountainous herbs. We gathered AFLP data and chloroplast DNA sequences (trnD‐trnT region) from 68 populations spanning the entire distribution range (the Jura Mountains, Alps, western Carpathians, Dinarides). Both genetic markers revealed two main phylogeographical groups (phylogroups) in C. purpurascens. Additionally, AFLP data detected a more detailed structure of five phylogroups: two widespread, showing east?west geographical separation, and three local ones, restricted to somewhat disjunct, marginal regions of the species range. We suggest that C. purpurascens survived the last glaciation in two main regions, the foothills of the Southern Limestone Alps and the Karst area of the north‐western Dinarides, and possibly also in microrefugia in the Western Carpathians. The glacial persistence and colonization routes of this woodland herb are highly concordant with those inferred for several temperate trees, especially the European beech. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 741–760.     

Subtle topographical differences along a floodplain promote different plant strategies among Paspalum dilatatum subspecies and populations

It was hypothesised that subtle topographical differences might cause the existence of ecotypes along a floodplain. The apomict grass Paspalum dilatatum subspecies dilatatum inhabits flood‐prone lowlands as well as nearby uplands in the floodplains of Argentina, while the sexual P. dilatatum subspecies flavescens almost exclusively inhabits the uplands. The aim of the present study was to identify the different traits that allow these P. dilatatum populations to inhabit different habitats. Plants of P. dilatatum were reciprocally transplanted between uplands and lowlands. Morphophysiological traits related to flooding tolerance were measured during a flood. Subspecies dilatatum from the uplands and subspecies flavescens showed a high physiological performance in the uplands but a considerable decrease in stomatal conductance, net photosynthesis rates and tiller number in the flooded lowlands. In contrast, the subspecies dilatatum from the lowlands showed relatively lower and stable stomatal conductance, photosynthesis rates and leaf water potential at both sites. Subspecies dilatatum from the lowlands outperformed upland populations at the lowland site with respect to tillering. Leaves of subspecies dilatatum from the lowlands that had grown at the lowland habitat had a lower blade/sheath proportion than leaves of plants transplanted to the uplands. This behavior did not occur in both upland populations. Results suggest that dilatatum Lowland plants have the typical strategy of stress‐tolerant genotypes and that the upland populations are adapted to habitats where competitive species are selected. In conclusion, habitats with subtle differences in topographic level can favour both ecotypic differentiations within an apomict subspecies but also the maintenance of morphophysiological similitudes between coexisting upland populations belonging to different subspecies.     

Clinal variation or validation of a subspecies? A case study of the Graptemys nigrinoda complex (Testudines: Emydidae)

Widely distributed species often display intraspecific morphological variation due to the abiotic and biotic gradients experienced across their ranges. Historically, in many vertebrate taxa, such as birds and reptiles, these morphological differences within a species were used to delimit subspecies. Graptemys nigrinoda is an aquatic turtle species endemic to the Mobile Bay Basin. Colour pattern and morphological variability were used to describe a subspecies (G. n. delticola) from the lower reaches of the system, although it and the nominate subspecies also reportedly intergrade over a large portion of the range. Other researchers have suggested that these morphological differences merely reflect clinal variation. Our molecular data (mtDNA) did not support the existence of the subspecies, as the haplotypes were differentiated by only a few base pairs and one haplotype was shared between the putative subspecies. While there were significant morphological and pattern differences among putative specimens of G. n. nigrinoda, G. n. delticola and G. n. nigrinoda × delticola, these differences probably represent clinal variation as they were also related to environmental variables [i.e. cumulative drainage area and drainage (categorical)]. Specimens occupying slow‐current, high‐turbidity river reaches (e.g. the Tensaw River) exhibited greater relative carapace heights and more dark pigmentation, while specimens occupying fast‐current, clearer rivers (e.g. the upper Alabama, Cahaba and Tallapoosa rivers) exhibited lower carapace heights and more yellow pigmentation. Given the absence of clear molecular and morphological differences that are related to drainage characteristics, we suggest that there is not sufficient evidence for the recognition of G. n. delticola as a distinct subspecies. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 810–822.     

Phylogeography of silver‐studded blue,Plebejus subsolanus (Lepidoptera: Lycaenidae), in the central mountainous regions of Japan

The silver‐studded blue, Plebejus subsolanus, is widely distributed in the Russian Altai mountains, northeastern China, the Korean Peninsula, and the Japanese archipelago. In Japan, the species is distributed across wide elevation ranges from the lowlands of Hokkaido to the subalpine zone of Honshu. Current subspecies classification in Japan is as follows: ssp. iburiensis, occurring in lowland grasslands in Hokkaido; ssp. yaginus in lower mountain grasslands in Honshu; and ssp. yarigadakeanus in higher mountain grasslands in Honshu. The habitat of this species has been markedly reduced due to recent habitat destruction and land‐use changes. Here, we undertook phylogeographic analyses of two subspecies, ssp. yaginus and yarigadakeanus in the central mountainous regions of Japan, based on two mitochondrial gene sequences, in order to collect information for establishing effective conservation strategies. From 57 samples from the four mountain ranges, we obtained a haplotype network comprised of 12 haplotypes. Because of the haplotype network topology, the geographic distribution of haplotypes and the correspondence of haplotype divergence to subspecies taxonomy, we provisionally divided the haplotypes into three haplogroups: YR1 and YR2, which comprised ssp. yarigadakeanus, and YG, which comprised ssp. yaginus. Mitochondrial DNA genetic differentiation generally agreed with morphological subspecies classification. The haplotype network suggested that ssp. yarigadakeanus populations had multiple origins, and the subspecies character of “bright blue of the male's wings” was assumed to have evolved independently in each subalpine meadow. We found that P. subsolanus was genetically differentiated depending upon the elevation at each mountain region, suggesting that each haplogroup should be a conservation unit.     

Phylogeography of the mole-shrew (Anourosorex yamashinai) in Taiwan: implications of interglacial refugia in a high-elevation small mammal

To test the Pleistocene interglacial refugia hypothesis with a high-elevation mammal, we studied the phylogeography of the mole-shrew (Anourosorex yamashinai) using partial mitochondrial cytochrome b gene sequences (737 bases). This shrew is endemic to Taiwan. It is mainly distributed in the highlands from 1000 to 2500 m in elevation. We examined 103 specimens from 24 localities in three mountain ranges of Taiwan and found 36 haplotypes. These haplotypes separated into two major phylogroups (Northern and Southern) plus a minor phylogroup (Houhuan) of only one haplotype. This demonstrated strong association with geography. The formation of these three phylogroups may be the result of interglacial refugia during the middle Pleistocene. Distinct sublineages were not found within each major phylogroup, suggesting that the populations (phylogroups) explosively expanded from the interglacial refugia of ancestral founder haplotypes. The present distribution pattern of haplotypes suggests that Mount Houhuan is an effective refugium in central Taiwan. It was not possible to specify the refugia for the Northern and Southern phylogroups.     

Mitochondrial DNA and plumage evolution in the white wagtail Motacilla alba

We analyzed sequences of two mitochondrial DNA (mtDNA) gene regions (control region and ND2) totaling 1477 base‐pairs from 232 specimens of the white wagtail Motacilla alba obtained from 27 localities throughout Eurasia. Although overall haplotype diversity was relatively low (0.79) and the most common haplotype was shared by 45% of individuals, belonging to six subspecies, a high level of population differentiation was detected. The mtDNA tree revealed three clades: (1) most individuals from Krasnodar (belonging to M. a. alba subspecies), (2) all individuals from Almaty and some from Primor'e (belonging to M. a. personata, M. a. lugens and M. a. leucopsis subspecies), and (3) the remaining individuals (representing all subspecies and all localities except Almaty). We suggest that these three clades represent historically isolated populations that relatively recently came into secondary contact in Krasnodar and Primor'e. None of the six subspecies were reciprocally monophyletic in the mtDNA tree. The Krasnodar population appeared to receive immigrants from other localities, but distinctive haplotypes from this locality did not appear elsewhere, suggesting asymmetric gene flow. Signatures of recent gene flow between northern populations were detected, and there was no evidence of isolation by distance within the northern group of populations. Mismatch distributions for most localities were consistent with population expansions. We also analyzed 12 male plumage characters from 93 study skins sampled from 24 populations. Phylogenetic trees resulting from separate genetic and morphological analyses were incongruent. Plumage evolution seems to be under strong sexual or natural selection, which favors particular phenotypes in various areas irrespective of the mitochondrial background. Dispersal events at different evolutionary times could have obscured the effects of earlier isolation events. The mtDNA data does not support species status for M. a. lugens and M. a. personata, which shared haplotypes with other subspecies of M. alba. We recommend that M. lugens and M. personata are placed as junior synonyms of M. alba.     

Home range and activity of the Indian giant flying squirrel (<Emphasis Type="Italic">Petaurista philippensis</Emphasis>) in Taiwan: influence of diet,temperature, and rainfall

Knowledge on home range and activity patterns, along with their responses to environmental fluctuations, is important for the understanding of wildlife ecology and conservation, but related studies on giant flying squirrel species (genus Petaurista) are still limited. We radio-tracked five Indian giant flying squirrels (Petaurista philippensis) in subtropical Taiwan to assess their home range and activity patterns, as well as their behavioral strategy to cope with fluctuations in food quality. Specifically, we assessed the travelling and resting times of P. philippensis in relation to its energy requirements during periods of low food quality in winter. The influence of temperature and rainfall was also investigated. A total of five individuals were radio-tracked for 1–6 months. The home ranges of four individuals averaged 2.8 ± 2.0 ha (± SD), based on the 95 % kernel method. Mean home ranges of two adult males (4.4 ± 1.3 ha) were larger than a female (1.8) and sub-adult male (0.8). P. philippensis was found to be more active around dusk and dawn and less active at midnight. Daily ranging distance and activities were negatively associated with proportion of mature leaves in diet of the only female that we tracked. Rainfall had negative effects on activities of the males, while temperature had no significant influence. The current study suggested an energy conservative strategy of P. philippensis. Home ranges of P. philippensis are smaller than those of smaller flying squirrel species (genus Glaucomys and Pteromys spp.), which may be related to the differences in food habits and gliding efficiency.     

Molecular phylogeography of European Sciurus vulgaris: refuge within refugia?

The red squirrel ( Sciurus vulgaris ) is a well-known forest animal distributed all over Europe. Still, we are far from having a firm knowledge of the species' phylogeography. This study investigates the genetic differentiation of S. vulgaris across the species' Eurasian range, using sequence data from the mitochondrial DNA gene (D-loop, 252 base pairs, cytochrome b , 359 base pairs), and eight variable autosomal microsatellite loci genotyped for 236 individuals. The results reveal the presence of two main mitochondrial phylogroups. The first clade comprises the individuals from the region of Calabria in southern Italy, belonging to the subspecies S. v . meridionalis , while the second clade contains the remainder of the studied individuals. Bayesian analysis of microsatellite genotypes resulted in three main clusterings corresponding to the three S. vulgaris subspecies: infuscatus, meridionalis and fuscoater . Geographical distribution of mtDNA haplotypes and mismatch analysis suggest a common refugium for the red squirrel across most of its present range from which expansion happened rather rapidly. The genotype mixing of italicus with northern populations could be a residual of postglacial expansion. The lack of mixing between the Calabrian lineage and the rest of European red squirrel haplotypes can be seen as evidence for distinct histories throughout the Pleistocene. Calabrian mtDNA probably diverged in an ice age contraction and remained isolated from the neighbouring squirrel populations until very recent times.     

Intra‐ and inter‐specific variation in size and habitus of two sibling spider species (Araneae: Lycosidae): taxonomic and biogeographic insights from sampling across Europe

Discriminating closely related species can become a taxonomical challenge if a clear morphological diagnosis is lacking. Two subspecies have been recognized in Pardosa agrestis (Westring, 1861); however, their validity is still debated. To resolve this dilemma, quantitative and qualitative traits were measured for 30 females and 30 males per site from ten localities throughout Western Europe (11 measurements on each individual) and compared between subspecies. Mean annual temperature and geographical coordinates from all sites were also included in the statistical models in order to test for variations in size over the distribution range of each subspecies. We found significant differences in body size and size of copulatory organs as well as accurate criteria of discrimination between P. a. agrestis and P. a. purbeckensis F.O.P. Cambridge, 1895, suggesting that these taxa are two valid morphological species, occurring sympatrically. We also showed that temperatures did not directly influence the size of individuals, but that one species had smaller individuals at higher latitudes (following the converse Bergmann's rule), and that both species had larger individuals in centrally sampled populations (fitting to the centre hypothesis). © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113, 85–96.