DNA barcoding of Murinae (Rodentia: Muridae) and Arvicolinae (Rodentia: Cricetidae) distributed in China

Identification of rodents is very difficult mainly due to high similarities in morphology and controversial taxonomy. In this study, mitochondrial cytochrome oxidase subunit I (COI) was used as DNA barcode to identify the Murinae and Arvicolinae species distributed in China and to facilitate the systematics studies of Rodentia. In total, 242 sequences (31 species, 11 genera) from Murinae and 130 sequences (23 species, 6 genera) from Arvicolinae were investigated, of which 90 individuals were novel. Genetic distance, threshold method, tree‐based method, online BLAST and BLOG were employed to analyse the data sets. There was no obvious barcode gap. The average K2P distance within species and genera was 2.10% and 12.61% in Murinae, and 2.86% and 11.80% in Arvicolinae, respectively. The optimal threshold was 5.62% for Murinae and 3.34% for Arvicolinae. All phylogenetic trees exhibited similar topology and could distinguish 90.32% of surveyed species in Murinae and 82.60% in Arvicolinae with high support values. BLAST analyses yielded similar results with identification success rates of 92.15% and 93.85% for Murinae and Arvicolinae, respectively. BLOG successfully authenticated 100% of detected species except Leopoldamys edwardsi based on the latest taxonomic revision. Our results support the species status of recently recognized Micromys erythrotis, Eothenomys tarquinius and E. hintoni and confirm the important roles of comprehensive taxonomy and accurate morphological identification in DNA barcoding studies. We believe that, when proper analytic methods are applied or combined, DNA barcoding could serve as an accurate and effective species identification approach for Murinae and Arvicolinae based on a proper taxonomic framework.     

Arboreal gaits in three sympatric rodents Apodemus agrarius,Apodemus flavicollis (Rodentia,Muridae) and Myodes glareolus (Rodentia,Cricetidae)

Vertical stratification of the arboreal habitat allows the coexistence of several species in a given area, because the complex arboreal strata can be used in different ways by arboreal and scansorial mammals. The present report experimentally investigated the gait metrics on different arboreal substrates, of three sympatric rodents living in a deciduous forest in Poznań, Poland. Arboreal locomotion was compared between the burrowing striped field mouse, Apodemus agrarius, the scansorial bank vole, Myodes glareolus, and the more arboreal yellow-necked mouse, Apodemus flavicollis. We filmed two wild-caught individuals from each species walking on four different substrate diameters (2 mm, 5 mm, 10 mm, 25 mm) and three different inclinations (45° descending, horizontal, 45° ascending) at 240 fps and collected a set of gait parameters from a total of 273 complete cycles. Our results did not demonstrate clear relationships between arboreal locomotion and the ecology of the three species. Only A. flavicollis exhibited locomotor features partly associated with arboreal competence, including lower velocity and diagonality on narrow substrates and asymmetrical gaits on wider ones. On the other hand, the two Apodemus species, despite their different ecologies, shared a few locomotor similarities, such as velocity regulation primarily by stride frequency, and similar effects of substrate size and inclination on diagonality, duty factor, and duty factor index indicating the possibility of a phylogenetic signal. Because the selected gait parameters provided limited insight into the ability of small mammals to move competently through an arboreal habitat, these findings indicate that the relationship between behaviour and ecology is complex.     

Hidden heterochromatin: Characterization in the Rodentia species Cricetus cricetus, Peromyscus eremicus (Cricetidae) and Praomys tullbergi (Muridae)

The use of in situ restriction endonuclease (RE) (which cleaves DNA at specific sequences) digestion has proven to be a useful technique in improving the dissection of constitutive heterochromatin (CH), and in the understanding of the CH evolution in different genomes. In the present work we describe in detail the CH of the three Rodentia species, Cricetus cricetus, Peromyscus eremicus (family Cricetidae) and Praomys tullbergi (family Muridae) using a panel of seven REs followed by C-banding. Comparison of the amount, distribution and molecular nature of C-positive heterochromatin revealed molecular heterogeneity in the heterochromatin of the three species. The large number of subclasses of CH identified in Praomys tullbergi chromosomes indicated that the karyotype of this species is the more derived when compared with the other two genomes analyzed, probably originated by a great number of complex chromosomal rearrangements. The high level of sequence heterogeneity identified in the CH of the three genomes suggests the coexistence of different satellite DNA families, or variants of these families in these genomes.     

Karyotypie diversity and taxonomic problems in the genus Arvicanthis (Rodentia,Muridae)

The analyses of R- and C-banding patterns of chromosomes of Arvicanthis niloticus originating from two different localities (Egypt and Central African Republic) revealed karyotypic differences caused by one pericentric inversion and three translocations, one being reciprocal and the others Robertsonian. There were also some differences in centromeric heterochromatin patterns.The data indicate that these two forms are distinct species, cytogenetically isolated, and that a revision of the taxonomic status of the genus Arvicanthis is needed.     

Morphological diversity of Old World rats and mice (Rodentia, Muridae) mandible in relation with phylogeny and adaptation

The respective roles of the phylogenetic and ecological components in an adaptive radiation are tested on a sample of Old World rats and mice (Muridae, Murinae). Phylogeny was established on nuclear and mitochondrial genes and reconstructed by maximum likelihood and Bayesian methods. This phylogeny is congruent with previous larger scale ones recently published, but includes some new results: Bandicota and Nesokia are sister taxa and Micromys would be closely related to the Rattus group. The ecological diversification is investigated through one factor, the diet, and the mandible outline provides the morphological marker. Elliptic and radial Fourier transforms are used for quantifying size and shape differences among species. Univariate size and shape parameters indicate that phylogeny is more influential on size than diet, and the reverse occurs for shape and robust patterns are recognized by multivariate analyses of the data sets provided by the Fourier methods. Omnivorous and herbivorous groups are well separated despite some overlapping, as well as are other Murinae with a specialized diet (insects, seeds). Phylogeny is also influential as shown by the segregation of several groups (Praomys, Arvicanthini, Rattus, Apodemus). Allometric shape variation was investigated, and although present it does not overwhelm effects of either phylogeny or diet. Massive mandibles characterize herbivorous Murinae and slender mandibles, the insectivorous ones. A strong angular process relative to the coronoid process characterizes seedeaters, and the reverse characterized Murinae with a diet based largely on animal matter. Such changes in morphology are clearly in relation with the functioning of the mandible, and with the forces required by the nature of the food: the need of a stronger occlusal force in herbivorous species would explain massive mandibles, and an increase of the grasping and piercing function of incisors in insectivorous species would explain slender mandibles.     

Inter- and intra-individual chromosome variability in Thamnomys (Grammomys) gazellae (Rodentia,Muridae) B-chromosomes and structural heteromorphisms

The present paper reports intra- and inter-individual variability related to the occurrence of numerous B-chromosomes in Thamnomys (Grammomys) gazellae, a species of African Climber rat belonging to the dolichurus group. The frequency of B-chromosomes in somatic and spermatogonial metaphases is investigated, together with their behaviour during meiosis. Moreover, G-banding makes it possible to identify a structural polymorphism resulting from a pericentric inversion in a large chromosome (no. 6). The distribution of the constitutive heterochromatin has been assessed by C-banding. The nucleolus organizer regions (NOR's) were located by means of silver staining in four chromosomal pairs (nos. 1, 2, 4, and 6). The karyotype of T. (G.) gazellae is compared with that of other taxa of the dolichurus group, particularly the Somaliland population which also exhibits the occurrence of B-chromosomes. The origin and significance of B-chromosomes is discussed.     

Structural Features of Spines in Some Rodents (Rodentia: Myomorpha, Hystricomorpha)

The architectonics of spines and hair was studied in Neacomys spinosus, Tokudaia osimensis, Arvicanthis somalicus, Leopoldamys sabanus, L. edwardsi, Maxomys moi, M. surifer, Niviventer fulvescens, N. confucianus, N. cremoriventer, Acomys cahirinus, A. somalica, Hystrix indica, H. cristata, Atherurus macrourus, Erethizon dorsatum, Proechimys steerei, and Lonchotrix emiliae. The presence of a dorsal longitudinal furrow covered by a modified cuticle is a common structural feature of spines in all studied species except E. dorsatum, H. indica, and H. cristata. The frontal surface of the mosaic (scales located side by side) and terrace cuticle (steps between scales) varies in different species from smooth to rib-folded. A terminology has been proposed to describe the main structures of spine and hair. The adaptive significance of the spine architectonics in rodents is discussed.     

Genome analysis of Peromyscus (Rodentia,Cricetidae)

A satellite DNA fraction from P. eremicus, having a buoyant density of 1.705 g/ml in neutral CsCl density gradients, was isolated. In situ hybridization experiments, using 3H-RNA complementary to this DNA fraction indicated that the short (heterochromatic) arms of most of the autosomes contained this sequence. Conversely, in situ hybridization using 3H-complementary DNA (cDNA) synthesized from the cytoplasmic poly (A) RNA of P. eremicus (comprising a substantial fraction of total messenger RNA) showed that the number of silver grains in the long arms (euchromatin) was significantly higher than that in the short arms. The X chromosomes showed a distinct localization pattern of both sequences.     

Discongruence of Mhc and cytochrome b phylogeographical patterns in Myodes glareolus (Rodentia: Cricetidae)

In the present study, a phylogeographical approach was developed to analyse the influence of selection and history on a major histocompatibility complex (Mhc) class II gene polymorphism in European bank vole (Myodes glareolus) populations. We focused on exon 2 of the Dqa gene because it is highly variable in a large array of species and appears to evolve under pathogen‐mediated selection in several rodent species. Using single‐strand conformation polymorphism analysis and sequencing techniques, 17 Dqa‐exon2 alleles, belonging to at least two different copies of Dqa gene, were detected over the distribution range of M. glareolus. Evidence of selection was found using molecular and population analyses. At the molecular level, we detected 13 codons evolving under positive selection pressures, most of them corresponding to regions coding for putative antigen binding sites of the protein. At the European level, we compared patterns of population structure for the Dqa‐exon2 and cytochrome b (cyt b) gene. We did not detect any spatial genetic structure among M. glareolus populations for the Dqa‐exon2. These results strongly differed from those obtained using the cyt b gene, which indicated a recent phylogeographical history closely linked to the last glacial events. Seven mitochondrial lineages have yet been described, which correspond to major glacial refugia. Altogether, our results revealed clear evidence of balancing selection acting on Dqa‐exon2 and maintaining polymorphism over large geographical areas despite M. glareolus history. It is thus likely that Mhc phylogeographical variability could have been shaped by local adaptation to pathogens. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 881–899.     

Host location,survival and fecundity of the Oriental rat flea Xenopsylla cheopis (Siphonaptera: Pulicidae) in relation to black rat Rattus rattus (Rodentia: Muridae) host age and sex

Host choice and fecundity are two factors that may contribute to the variation in flea counts observed when assessing the potential risk of flea-borne transmission of pathogens from rodents to humans. Using the black rat, Rattus rattus Linnaeus, as host the effects of age and sex on host choice and fecundity of the Oriental rat flea, Xenopsylla cheopis Rothschild, were examined experimentally at 25 degrees C and 80% rh. During the first two days of emergence from cocoons, female fleas dominated the sex ratio by 4:1 but from the third day onwards this switched to a male-dominated sex ratio of 4:1. The sex of the flea did not influence their host-seeking behaviour. Newly emerged fleas of both sexes were not influenced by the rat's presence and at seven days old both sexes demonstrated similar levels of attraction toward the rat host. The sex of the rat did not affect flea host-seeking behaviour. There was a 50-70% decline in the initial number of adult fleas during the first week after their release onto a rat host, and this decline was greatest on juvenile rats. Flea fecundity was also significantly lower on juvenile rat hosts but no differences due to the sex of the rat were observed. This experimental study supports the hypothesis that differences in flea count due to host sex, reported in field surveys, result from sexual differences in host behaviour and not from discriminatory host-seeking behaviour by X. cheopis. Differences in flea count due to host age may be affected by differences in X. cheopis fecundity, which may itself be mediated by host behaviour such as grooming.     

Unusual C-band patterns in three karyotypically rearranged forms of Scapteromys (Rodentia, Cricetidae) from Brazil

Chromosome studies of 30 specimens of the rodent Scapteromys collected at nine localities in southern Brazil revealed the occurrence of three karyotypic taxa with 2n = 36 (one locality), 34 (two localities), and 24 (six localities), although all three had 40 autosomal arms (AN). The G-band analysis indicated that this reduction in diploid number was mainly due to Robertsonian translocations which have occurred along a gradient, possibly in two independent evolutive routes. The C-bands occur on one autosomal pair and on the X and Y in the 2n = 36 and 34 forms and on the X and Y chromosomes only in the 2n = 24 taxon. The broad genomic reorganization which has occurred in this genus, in which the chromosomes do not have large amounts of constitutive heterochromatin, argues against the idea that a large amount of constitutive heterochromatin favors chromosome evolution and speciation.     

Bibliography of the genus Apodemus (Rodentia, Muridae)

A complete bibliography of the genus Apodemus for the period 1970–1980 is presented, with a selection of the more important papers from earlier years, and a few references for 1981. A coded index to species and subjects is provided. All titles have been translated into English.     

Reproductive characteristics of free-livingCalomys venustus (Rodentia, Muridae)

Calomys venustus showed a cyclic seasonal pattern of reproductive activity characterized by a period of repose and a variable length of the period of sexual activity between August—September to May—June. The study was done from 1983 to 1999. During this time the population density increased from October—November to the end of autumn and sharply decreased in winter. Three peaks in rate of pregnancy were recognized in the breeding period: spring, summer and late summer. The average litter size was different among the three peaks in rate of pregnancy. The reproductive pattern had a peak at the beginning of the spring produced by overwintering animals, and another peak in summer-autumn caused by the reproductive activity of young born in the same period in which they breed. The spring cohort exhibited the greatest breeding contribution suggesting that this cohort is principally responsible for the yearly peak in abundance.     

Calomys laucha (Rodentia, Cricetidae): growth and breeding in laboratory conditions

The husbandry and breeding of Calomys laucha (Rodentia, Cricetidae) in captivity are described. Growth curves based on body weight and length showed statistical differences between sexes after 45 days, males being heavier than females. The overall reproductive efficiency was 53.4% but birth rate was depressed during winter. Gestation length was 21 +/- 1 days and females exhibited postpartum oestrus with a 3-7 day implantation delay (51%). Litter size was 5.3 +/- 1.1 (n = 34). Pup survival at weaning was 84.9%. Mean life span in laboratory conditions was 13.5 months and a cumulative mortality of 90% was reached at 27-28 months of age.     

Genetic diversity and phylogenetic relationships in the zokor subfamily myospalacinae (Rodentia,Muridae) inferred from RAPD-PCR

Zokors (Myospalacinae) is a group of rodents specialized for underground life, endemics of eastern Asia, which is taxonomically and evolutionarily poorly studied. We examined genetic diversity and phylogenetic relationships among zokors (Myospalax myospalax, Myospalax aspalax, Myospalax armandii, Myospalax psilurus, Myospalax smithii) using RAPD-PCR. The subfamily Myospalacinae was shown to be monophyletic and contain four evolutionary branches: M. myospalax, M. aspalax-M. armandii, M. smithii and M. psilurus. Genetic differences and high differentiation were found among the species and between two geographic forms of Manchurian zokor M. psilurus from the marginal parts of the range, Transbaikalia and Primorye. The psiluris phylogroup was shown to be dichotomically divided into two clades according to the geographical distribution of animals from Transbaikalia and Primorye. The genetic differentiation between the geographic forms of M. psilurus corresponded to the differentiation between morphologically similar species M. aspalax and M. armandii. M. armandii is a sister taxon with regard to M. aspalax. This new evidence on the evolutionary relationships among zokors does not contradict the traditional views inferred from morphological, karyological, and allozyme data, on apartness of M. myospalax and the character of evolution in this group. The species status of Myospalax psilurus Milne-Edwards, 1874, M. epsilanus Thomas, 1912, and M. armandii Milne-Edwards, 1867, which had been suggested earlier on the basis of biochemical and karyological data, was confirmed.     

Characterization of two distinct species of Arvicanthis (Rodentia: Muridae) in West Africa: cytogenetic, molecular and reproductive evidence

The unstriped grass rat, Arvicanthis Lesson 1842, is one of the most common genera of murid rodents in African savannas. However, from a systematic viewpoint, very little is known about this group. Following recent investigations which showed karyotypic variability within the species A. niloticus , the present study attempts to clarify the nature and distribution of these chromosomal variants, as well as to determine their taxonomic rank.
The chromosomes of 15 individuals from different West African localities were prepared from fibroblast cultures, and R- and C-banded karyotypes were constructed. In addition, the levels of genetic divergence (DNA/DNA hybridization) and reproductive isolation (attempted crossbreeding in captivity) were examined. The results confirm the existence of two differentiated karyomorphs, differing by numerous chromosomal rearrangements such as pericentric inversions and translocations, as well as differences in the quantity of constitutive heterochromatin. These karyomorphs appear to be genetically and reproductively isolated and are parapatrically distributed; their areas of distribution correspond to the sahelian and sudano-guinean domains, respectively. The distinctness of these karyomorphs, the absence of hybrids in laboratory crosses, and the pronounced genetic divergence provide good evidence for the recognition of two distinct sibling species. We propose to keep the designation A. niloticus for the northern sahelian form and discuss the naming alternatives for the other.