Ongoing ultradian activity rhythms in the common vole,Microtus arvalis,during deprivations of food,water and rest

Summary The timing mechanism underlying ultradian (2–3 h) activity patterns in the common vole, Microtus arvalis, was studied using behavioural deprivation experiments. These were aimed at distinguishing between a homeostatic control mechanism, in which the rhythmic behaviour itself is part of the causal loop, and a clock mechanism, independent of the behaviour.In 175 experiments, deprivation of food during 3 ultradian cycles in (subjective) daytime did not result in significant changes in the ultradian periodicity of attempts to obtain the food, compared with ad lib. access to food and water. A minor, but significant increase in ultradian activity time () occurred in the course of the deprivation, but this was compensated by a shorter ultradian rest (). These results were obtained both in intact animals (n = 24), which showed ultradian and circadian rhythmicity in behaviour, and in animals (n = 21) with electrolytic lesions aimed at the suprachiasmatic nuclei (SCN), which lacked the circadian modulation of behaviour. Simultaneous deprivation of water and food in 8 voles without circadian rhythmicity during 40 experiments also did not lead to any change in the ultradian periodicity of feeding attempts.Rest deprivation was studied in 5 SCN lesioned voles, by forcing running wheel activity to continue following spontaneous running. Thus, the experimental activity bout was artificially lengthened to 2–9 h in 67 experiments. The onset of the subsequent rest episodes occurred independent of the duration of the preceding . The duration of was dependent on the preceding, experimental in a periodic fashion. The interval experimental (=lengthened +following ) was equal to one, two or three times the control (obtained on nonexperimental days). This result fits the prediction of a clock model and is in conflict with a monotonicincrease of with , as expected in a homeostatic, restorative process.It is concluded that the ultradian timing of activity in the common vole can be explained neither by homeostatic hunger or thirst mechanisms nor by homeostatic rest/activity regulation. The results strongly suggest an independent clock system generating ultradian feeding rhythms in the common vole.Abbreviations DD continuous darkness - LD light-dark regime - LL continuous light - RCA retrochiasmatic area - ARC arcuate nucleus - SCN suprachiasmatic nuclei - ultradian period - ultradian activity time - ultradian rest time     

Differential elimination of circadian and ultradian rhythmicity by hypothalamic lesions in the common vole, Microtus arvalis

Effects of hypothalamic lesions on the ultradian and circadian organization of wheel running and feeding were studied in the common vole, Microtus arvalis. Circadian organization broke down within 30 days in continuous darkness in 24% of intact voles (n = 135). Ultradian rhythmicity of feeding (period 2-3 hr) persisted in constant conditions in all intact voles. Following lesions of the suprachiasmatic nuclei (SCN), circadian rhythmicity disappeared when lesions were complete (n = 8) or more extensive than 25% of the total SCN volume (n = 5). Absence of circadian rhythmicity was also found in animals with substantial lesions in the diencephalic paraventricular area (PVA) and in the retrochiasmatic area (RCA) and/or adjacent arcuate nucleus (Arc). Complete loss of ultradian and circadian organization occurred in eight voles with damage to the RCA and/or Arc. In three of these, the SCN was intact. The SCN is a likely candidate for a circadian pacemaker in voles (as in other rodents), while the loss of circadian rhythmicity following PVA and RCA/Arc lesions may be due to destruction of efferent pathways from the SCN. The RCA/Arc area is apparently necessary for the expression of ultradian rhythms. The intact SCN is neither necessary nor sufficient for the generation of ultradian rhythmicity.     

Fine-scale genetic structure and dispersal in the common vole (Microtus arvalis)

The genetic structure and demography of local populations is tightly linked to the rate and scale of dispersal. Dispersal parameters are notoriously difficult to determine in the field, and remain often completely unknown for smaller organisms. In this study, we investigate spatial and temporal genetic structure in relation to dispersal patterns among local populations of the probably most abundant European mammals, the common vole (Microtus arvalis). Voles were studied in six natural populations at distances of 0.4-2.5 km in three different seasons (fall, spring, summer) corresponding to different life-history stages. Field observations provided no direct evidence for movements of individuals between populations. The analysis of 10 microsatellite markers revealed a persistent overall genetic structure among populations of 2.9%, 2.5% and 3% FST in the respective season. Pairwise comparisons showed that even the closest populations were significantly differentiated from each other in each season, but there was no evidence for temporal differentiation within populations or isolation by distance among populations. Despite significant genetic structure, assignment analyses identified a relatively high proportion of individuals as being immigrants for the population where they were captured. The immigration rate was not significantly lower for females than for males. We suggest that a generally low and sex-dependent effective dispersal rate as the consequence of only few immigrants reproducing successfully in the new populations together with the social structure within populations may explain the maintenance of genetic differentiation among populations despite migration.     

Unusual heteromorphic bivalents in the common vole (Microtus arvalis) from Belorussia.

Electron microscopic analysis was carried out on the synaptonemal complexes of 10 male common voles (Microtus arvalis) caught in 1990 in Belorussia. In the early pachytene stage of spermatocytes of four males, a heteromorphic bivalent has been found in one of five large autosomes. In the central region of the bivalent one of the lateral elements is in the form of a D-loop, characteristic of insertion/deletion heterozygotes. However, high-resolution G-band staining of mitotic chromosomes from fibroblasts shows no significant differences in the G-band pattern between homologs.     

Possible atypical course of tularemia (persistence) in the common vole Microtus arvalis Pall

The possibility of the atypical course of tularemia with the prolonged persistence of Francisella tularensis in common voles (M. arvalis), the twin species of East European voles (M. rossiaemeridionalis), was studied. Experiments were made on 33 animals grown in the laboratory. F. tularensis strain 165 was used. The animals were infected by feeding them according to the previously developed scheme. 7 out of 33 voles showed the atypical course of tularemia: in 3 voles the disease took a prolonged course with bacteriuria and death on days 25-34; 3 other voles with bacteriuria registered before days 33, 66 and 172 (the term of observation) survived. The surviving animals were killed on day 183, and the presence of bacteria in their organs and seroconversion were established. One vole excreted no bacteria with urine and had no bacteria in its organs (the animal was examined on day 156), but in its blood specific antibodies were detected. To determine bacteriuria, the immunofluorescence test was used together with biological assays. Thus, M. arvalis, like M. rossiaemeridionalis studied earlier, can harbor F. tularensis at the period between epizootics. When voles of the former species penetrate stacks of straw and hayricks, conditions appear for the transfer of the infection to the latter species, M. rossiaemeridionalis. Therefore, in the foci of the meadow-field type each of these two species of voles may be not only of epizootic, but also of epidemic importance.     

Polymorphic microsatellite loci and PCR multiplexing in the common vole,Microtus arvalis

We isolated and characterized 14 dinucleotide microsatellite loci in the common vole Microtus arvalis (Palas). Two multiplex panels both comprising seven loci were developed. Application to a set of 21 individuals allowed clear and easy characterization of allele sizes except for two loci which were then withdrawn from further analyses. The number of alleles ranged from four to 19 per locus with the observed heterozygosity ranging from 0.55 to 0.95. These sets of microsatellite loci provide high throughput capacity for population genetic studies at a minimum cost.     

Transalpine colonisation and partial phylogeographic erosion by dispersal in the common vole (Microtus arvalis)

The colonisation history and genetic structure of the common vole ( Microtus arvalis ) was investigated in the region of the Alps by analysing the mitochondrial cytochrome b gene (mtDNA) and 19 microsatellite loci (nucDNA) for 137 voles from 52 localities. mtDNA data provided a much refined distribution of three highly divergent evolutionary lineages in the region compared to previous studies. Although high mountain ranges are widely accepted to be barriers for colonisation processes for many organisms and especially small terrestrial mammals, our phylogeographic analyses showed clear evidence of four transalpine colonisation events by the common vole. Individual-based phylogenetic analyses of nucDNA and two alternative Bayesian-clustering approaches revealed a deep genetic structure analogous to mtDNA. Incongruence between nucDNA and mtDNA at the individual level was restricted to the regions of contact between the lineages. mtDNA patterns and strong female philopatry in M. arvalis suggest that the crossings of the Alps occurred during the colonisation of the region when it was free from ice after the last glaciation. nucDNA patterns suggest that some of the transalpine elements of this phylogeographic pattern were subsequently eroded by male-biased gene flow. We conclude that the combination of phylogeography and landscape genetics at the individual level can provide very detailed insights into colonisation events and may even allow differentiation between historical and more recent processes.     

多位点DNA指纹技术在保加利亚普通田鼠中的应用探讨

DNA指纹是一种重要的现代分子遗传学标记技术（Jeffreys et al．,1985）,它所揭示的是生物体大量的、无遗传编码信息的、具有高度多态性的卫星DNA（Chen,1996）。这些DNA序列往往占据了生物体基因组总量的80％以上,由于它不编码蛋白基因,在系统发育过程中,通常不被自然选择和人工选择,使得生物变异积累形成个体基因组间的巨大差异。因此,DNA指纹受到生物学家的青睐,以用于生物个体和群体的基因组分析（Burke and Bruford,1987;Buitmap et al．,1991;Weising et al．,1995）。     

Genetic structure and colonization processes in European populations of the common vole, Microtus arvalis

The level of genetic differentiation within and between evolutionary lineages of the common vole (Microtus arvalis) in Europe was examined by analyzing mitochondrial sequences from the control region (mtDNA) and 12 nuclear microsatellite loci (nucDNA) for 338 voles from 18 populations. The distribution of evolutionary lineages and the affinity of populations to lineages were determined with additional sequence data from the mitochondrial cytochrome b gene. Our analyses demonstrated very high levels of differentiation between populations (overall FST: mtDNA 70%; nucDNA 17%). The affinity of populations to evolutionary lineages was strongly reflected in mtDNA but not in nucDNA variation. Patterns of genetic structure for both markers visualized in synthetic genetic maps suggest a postglacial range expansion of the species into the Alps, as well as a potentially more ancient colonization from the northeast to the southwest of Europe. This expansion is supported by estimates for the divergence times between evolutionary lineages and within the western European lineage, which predate the last glacial maximum (LGM). Furthermore, all measures of genetic diversity within populations increased significantly with longitude and showed a trend toward increase with latitude. We conclude that the detected patterns are difficult to explain only by range expansions from separate LGM refugia close to the Mediterranean. This suggests that some M. arvalis populations persisted during the LGM in suitable habitat further north and that the gradients in genetic diversity may represent traces of a more ancient colonization of Europe by the species.     

A comparison of methods for estimating common vole (Microtus arvalis) abundance in agricultural habitats

Rodent outbreaks cause significant crop damages in agricultural areas worldwide, but routinely monitoring large areas at low cost remains a challenge. The common vole Microtus arvalis has recently colonized the agricultural plains of the northern Iberian Plateau, an area where it has started to produce population outbreaks with important impacts in agriculture, the environment and human health. Vole monitoring has become of prime importance to implement preventive management measures to control populations. In order to find a simple and reliable vole monitoring method to be applied in large areas, we compared abundance estimates derived from three methods: capture–mark–recapture (CMR), single capture events (SCE) and presence/absence of vole activity signs (VAS) during three seasons and on the main agricultural habitats in the study area. We show that an activity index based on the presence of fresh droppings and/or clippings had a similar performance to SCE in a large sample of plots (n = 222) across habitats and seasons. Data obtained with both methods (SCE, VAS) were also well correlated with those obtained with CMR, despite a limited sample size (n = 23 CMR plots). We suggest that the VAS method, which is a cheaper and easier alternative to trapping methods, provides a promising tool for scientists and managers to implement large scale monitoring of common vole in agricultural areas.     

Intrapopulation autosomal polymorphism in the common vole Microtus arvalis from the Transcaucasian region

The broad autosomal polymorphism in form obscurus of common voles Microtus arvalis from the Transcaucasian region that is associated with the variation of subtelocentric chromosome pair 5, as well as the mechanism and evolutionary significance of this polymorphism, are discussed. Based on the morphological analysis of heterozygotes for chromosome pair 5 after differential G-, C-, and Ag-NOR-banding and on the measurements of homologues, the following conclusion has been made. The occurrence of the acrocentric chromosome 5 is the result of a double chromosomal rearrangement: a pericentric inversion and a duplication of the chromosomal material. The mutation has been found throughout the entire territory of Armenia. In spite of such a wide distribution, the mutation frequency in populations is extremely low. Neither a definite pattern of geographic distribution nor a clinal variation was found for this mutation. This mutation is likely to occur independently in different M. arvalis populations and is apparently neutral. Homozygotes for chromosome pair 5 are described for the first time.     

Characteristics of tear proteins in the vole, Microtus arvalis

Tear proteins were detected by polyacrylamide gel electrophoresis in the vole, Microtus arvalis. The tear proteins were separated to 6 to 8 bands and the bands were divided to three regions on the anodic side. In the adult male vole, a male specific band (Vtp-1) was detected in the first region. The first region of adult female and immature voles contained two specific bands (Vtp-2, 3). In the castrated adult males or adult males injected with estrogen, the male specific hand, Vtp-1, disappeared and Vtp-2 and 3 bands appeared. In all castrated voles, the Vtp-1 band appeared and Vtp-2 and 3 bands disappeared after the administration of testosterone. Thus, sex hormone-dependent proteins are present in vole tears.     

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.