We sequenced 704 mitochondrial DNA (mtDNA) control-region nucleotides and genotyped 11 autosomal microsatellites (STR) in 617 European roe deer (Capreolus capreolus) samples, aiming to infer the species' phylogeographical structure. The mtDNA sequences were split in three distinct haplogroups, respectively, named: Clade West, sampled mainly in Iberia; Clade East, sampled mainly in Greece and in the Balkans; and Clade Central, which was widespread throughout Europe, including the eastern countries and Iberia, but not Greece. These clades might have originated in distinct Iberian and Balkanic refuges during the penultimate or the last glaciations. Clades East and West contributed little to the current postglacial mtDNA diversity in central Europe, which apparently was recolonized mainly by haplotypes belonging to Clade Central. A unique subclade within Clade Central grouped all the haplotypes sampled from populations of the Italian subspecies C. c. italicus. In contrast, haplotypes sampled in central and southern Spain joined both Clade Central and Clade West, suggesting that subspecies C. c. garganta has admixed origin. STR data support a genetic distinction of peripheral populations in north Iberia and southern Italy, and show the effects of anthropogenic disturbance in fragmented populations, which were recently reintroduced or restocked and not may be in mutation-drift equilibrium. Roe deer in central Europe are mainly admixed, while peripheral populations in north Portugal, the southern Italian Apennines and Greece represent the remains of refugial populations and should be managed accordingly.

a b s t r a c tThe focus of studies of sexual dimorphism has been mainly on size whereas shape has remained largely ignored when assessing population differentiation. In order to extract exclusive information about shape variability, we combined multivariate statistical procedures with geometric morphometrics and visualization techniques (thin plate splines). All roe deer skulls, used in this study, originated from a single population in Serbia. Our results showed statistically significant sexual dimorphism of male and female skull shape. Different patterns of shape variability in males and females were observed, such as broader cranial base in males and elongated and more slender shape of female skull, with narrower basicranium. Shape differences appear to be regulated only broadly by social structure and sexual selection, and some other mechanisms could be proposed for further explanation of sexual dimorphism.

Complex evolutionary interactions can cause differential responses of males and females to environmental factors which result in variations of the degree of sexual dimorphism across different habitats. Roe deer Capreolus capreolus is an excellent model species for analyzing sexual dimorphism in the context of habitat variability as the most widespread ungulate species in Europe. The impact of three different habitat types (closed, intermediate and open) on the level of cranial integration in roe deer and patterns between sexes was tested by analyzing 761 adult craniums from 11 roe deer populations in Serbia. Our results confirmed higher level of integration and more pronounced sexual dimorphism in closed habitats in comparison with open habitats. Males also showed different patterns of integration across habitats than females. The general consistency of results across different tests suggests that patterns of integration between sex and habitat groups tend to be different for males and females from different habitat types. When faced with strong selective pressures, patterns of correlations among skeletal elements can evolve even within a species as an indirect influence of social organization through habitat and sexual selection. We propose that cranial integration in roe deer evolved according to the predictions of the adaptive model of phenotypic differentiation within a taxon in closed habitats channeled by stabilizing selection. The different patterns of cranial integration between sexes after successful colonization of intermediate and open habitats can be explained by a change in overall selective pressures to disruptive/ directional selection, thus breaking up observed patterns of integration, since they are treated as a constraint in changed circumstances.

Miloševiae-Zlatanoviae S., Crnobrnja-Isailoviae J. and Stamenkoviae S. 2005. Allozyme variability and differentiation in Serbian roe deer populations Capreolus capreolus. Acta Theriologica 50: 429-444.The present study investigates the genetic structure of 12 roe deer Capreolus capreolus Linnaeus, 1758 population samples from Serbia, by screening a total of 334 individuals. We examined whether genetic differentiation exists in local populations in Serbia, and addressed the question whether management policies may affect genetic structure. The populations were analysed by multilocus protein electrophoresis, with 33 protein loci examined. Screening of 20 enzymes and one group of general proteins revealed polymorphism at the following 12 loci: Sdh, Mdh-1, aGpd, Ak, Ca, Mpi and Gpi. Among samples, the proportion of polymorphic loci varied between 3-15.2% (mean 11.9%), while the average gene diversity was in the range of 1.1-4.2%. The overall genetic differentiation was low (q = 0.03). The comparison of two regional population groups (northern-southern, separated by the Danube River) showed an absence of genetic differentiation between regions. Gene flow was estimated at 8.96 migrants per generation, and was higher in the lowland than in the highland group. Three loci (Ca, 6-Pgd and Gpd-1) showed clinal variation along a geographical gradient. Additional five alleles of four loci (Ak, Pgm-1, Gpi, 6-Pgd) showed significant spatial autocorrelation. Genetic distances were small (D = 0-0.004). Northern and southern populations clustered separately. For at least three populations game management practices provide evidence for outlying genetic parameters. The observed heterogeneity in the inbreeding level was deemed more under the influence of non-random mating strengthened by game management, than by overall selective pressure. IntroductionThe results of previous studies reveal that genetic variability at the allozyme level is of the same order of magnitude in all examined populations of the roe deer, Capreolus capreolus Linnaeus 1758 Reimoser 1988, Hartl et al. 1993, [429] Acta Theriologica 50 (4): 429-444, 2005.PL ISSN 0001-7051 Lorenzini et al. 1993, Hewison 1995, Wang and Schreiber 2001 in Europe. In addition, almost the same loci are polymorphic in the different European populations. Selection of genes, number of populations and sample sizes, as well as modifications of the electrophoretic method, can influence the results and complicate comparisons (Simon and Archie 1985). However, it is also clear that the final pattern of the distribution of genetic variability can be strongly affected by many geographic, historical and demographic factors, as well as by those related to breeding structure, dispersal and the influence of anthropogenic factors (Fakler and Schreiber 1997, Lorenzini et al. 1997, Hartl et al. 1998. Recently, these studies have been expanded to include phylogeography across the distribution range in Europe using autosomal microsatellite loci (Randi et al. 2004) and mtDNA control regions Tiedem...

Diverse recombinant immunoreagents specific for TNT-derivatives were tested in different assay forms in order to analyze their specificity and sensitivity. Performance of immunoassays was based on TNP-protein conjugates immobilization on a solid surface. In this work, the detection limit for TNT-analog TNP-Tris was 250 fmol or 87 pg mL−1 (87 ppt), which represents the most sensitive assay published until now, regarding the detection of recombinant antibodies.