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Map showing sampling localities of grey wolves from Serbia. Geographic regions are marked as follows: I—Northern Serbia ; II—Western Serbia; III—Eastern Serbia; IV—Southern Serbia.
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Results of previous morphometric and genetic analyses of grey wolf (Canis lupus L.) population from Serbia indicated different patterns of population subdivision. In order to explore population structure, level of genetic variability, genetic drift, inbreeding and signals of bottleneck for grey wolves from Serbia, we applied highly polymorphic gene...
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... total number of 75 grey wolf muscle tissue samples from different localities across Serbia ( Fig. 1) was collected, during regular hunts. After sampling, tissues were preserved in 95% ethanol and stored at -20°C. Total DNA was isolated using a phenol-chloroform method by Sambrook and Russel ...
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... A posteriori analysis of the STRUCTURE results for the com bined pool of the wolf and dog genotypes revealed the maxi mum value of the test statistics ΔK at K = 2, which indicates the presence of two genetic clusters in the analyzed sample of animals: the Grey wolf (green cluster) and the domestic dog (red cluster) ( Fig. 1, Supplementary Material 2). When determining the population structure of samples of wolves and dogs separately from each other, the cluster formed by the wolf samples remained homogeneous; the absence of cluster ing within the wolf population using STR loci was previously shown by researchers in Europe (Aspi et al., 2006;Sastre et al., 2011;Ðan et al., 2016). ...
Commercial panels of microsatellite (STR) loci are intended for DNA analysis of the domestic dog ( Canis lupus familiaris ) and, therefore, when genotyping the Grey wolf ( Canis lupus lupus ), most markers reveal significant deviations from the Hardy–Weinberg equilibrium and have a low informative value, which complicates their use in a forensic examination. The aim of this study was to select STR markers that equally effectively reflect population polymorphism in the wolf and the dog, and to create a universal panel for the identification of individuals in forensic science. Based on the study of polymorphisms of 34 STR loci, a CPlex panel of 15 autosomal loci and two sex loci was developed, which is equally suitable for identifying wolfs and dogs. Analysis of molecular variance (AMOVA) between samples revealed significant differentiation values ( F ST = 0.0828, p < 0.05), which allows the panel to be used for differentiating between wolf and dog samples. For the first time in the forensic examination of objects of animal origin in the Republic of Belarus, population subdivision coefficients (θ-values) were calculated for each of the 15 STR loci of the test system being reported. It was shown that the values of the genotype frequency, when averaged over all studied animals without and with considering the θ-value, differ by three orders of magnitude (3.39 · 10 –17 and 4.71 · 10 –14 , respectively). The use of population subdivision coefficients will provide the researcher with the most relevant results of an expert identification study. The test system was validated in accordance with the protocol of the Scientific Working Group on DNA Analysis Methods. A computational tool was developed to automate the analysis of genetic data on the wolf and dog in the forensic examination; two guides were approved for practicing forensic experts. This methodology is being successfully used in expert practice in investigating cases of illegal hunting, animal abuse and other offenses in the Republic of Belarus.
... A ma már CITES (Convention on International Trade in Endangered Species of Wild Fauna and Flora) listán szereplő, Európában kilenc állam kivételével védett, ill. fokozottan védett állatként nyilvántartott faj [3], a védelmüknek köszönhetően stabilizálni tudta megmaradt populációit, és elkezdte rekolonizálni hajdani területeit [4,5]. Napjainkban az európai farkasállomány lassan növekvő tendenciát mutat. ...
... Bár az mtDNS csak anyai ágon öröklődik, így egyedüli alkalmazása nem ad teljes képet hibridizációs kérdésekben, a KR vizsgálata mégis hasznos módszernek bizonyult későbbi kutatásokban [6,[53][54][55]. Pontosabb eredményeket hoztak a biparentális mikroszatellitákat használó vizsgálatok, amelyeket vagy önmagukban [28,56,57], vagy uniparentális markerekkel kiegészítve alkalmaznak [4,58]. A két markertípus együttes használata a korábbiaknál megbízhatóbb és pontosabb eredményt ad, így manapság is javasolt közel rokon taxonok megkülönböztetésére, valamint a hibridek detektálására [18,19,23,24,59,[60][61][62][63][64][65][66][67][68][69][70][71][72][73][74][75]. ...
ÖSSZEFOGLALÁS A szerzők a kutyák és farkasok különböző genetikai markerekkel történő elkülönítésének lehetőségeit vizsgálják és bemutatják ennek fontosságát az igazságügyi alkalmazás területén. A rendelkezésre álló farkas-és kutyaeredetű mintákból meghatározott mitokondriális kontrollrégió haplotípusok és a 14 vizsgált mikro-szatellita-alléleloszlás adatai alapján különbség látható a farkas-és kutyaminták között. Az eddigi hazai eredmények is alátámasztják annak lehetőségét, hogy különböző genetikai markerek párhuzamos vizsgálatával − amelyek megfelelnek az igazságügyi célú alkalmazás kritériumainak −, kellő valószínűséggel alátámasztható egy kérdéses eredetű minta alfajszintű besorolása.
SUMMARY
Background: After several decades of absence, the grey wolf (Canis lupus) has started recolonizing its former territories in Hungary at the beginning of the 21 st century. Due to the intense presence of mankind, wolves are forced to share great areas of land with humans, which potentially leads to several conflicts. From the wolves' perspective, it means the decimation of domestic livestock. As far as humans are concerned, these conflicts may manifest in the illegal hunting of wolves and trading with their products. When facing such case, it should be examined whether the perpetrator/victim is a wolf or a dog. Objective: The aim of our study was to test genetic methods which can be used for forensic application as well to distinguish between wolves, dogs, or their hybrids. Materials and Methods: Altogether 22 samples (hair, skin, faeces, saliva, and purified DNA) from wolves and wolf-dog hybrids were collected. For the comparative canine database DNA samples from Hungarian dog populations were used. After DNA isolation, the mitochondrial hypervariable region I (HVI) and 14 autoso-mal microsatellite markers were amplified by PCR (Polymerase Chain Reaction). Mitochondrial haplotypes determined by sequencing were grouped using PopART. Genetic profiles based on the detected microsatellite alleles were analysed using Structure 2.3.4 and were grouped based on a Bayesian approach. Results and Discussion: The mitochondrial control region (HVI) haplotypes were successfully determined from the examined samples; these sequences were uploaded to the GenBank database. We did not find similar point mutation patterns between wolves and dogs. However, difference between wolf and dog groups was shown based on the detected microsatellite allele distribution, to make the results even more reliable further markers and more wolf samples should be involved. Overall, our preliminary results support that simultaneous application of large number of genetic markers meeting the standards of forensic application criteria-, could be adequate to determine the precise taxonomic origin of questionable samples.
... Our results provide insights into the genetic diversity and population structure of wolves from Bosnia and Herzegovina, based on the largest sample size and area coverage. Moderately high genetic diversity based on heterozygosities was observed for our sample of wolves from Bosnia and Herzegovina, as observed for other Dinaric-Balkan wolf populations (Djan et al. 2016;Lucchini et al. 2004;Fabbri et al. 2014;Moura et al. 2014). A broad study conducted by Hindrikson et al. (2016) revealed the highest heterozygosities for the Dinaric-Balkan population in Europe. ...
... It is evident from the available scientific literature that the genetics of local subpopulations of the Dinaric-Balkan wolf population have mostly been explored in isolation, e.g. genetic structure based on microsatellite variability was explored in grey wolves from Croatia (Fabbri et al. 2014) separately from those of Serbia (Djan et al. 2016). Unlike the structuring observed for grey wolves in Croatia, no genetic structure was detected for grey wolves from Serbia. ...
The grey wolves of Bosnia and Herzegovina form a subpopulation of the Dinaric-Balkan wolf population and represent one of Europe’s least studied wolf populations. Since the Dinaric-Balkan population is a valuable source of genetic diversity for neighboring populations, comprehensive assessments are warranted. We aimed to determine the genetic variability and structure of the grey wolf population from Bosnia and Herzegovina, as well as estimate levels of gene flow and inbreeding and evaluate genetic signals of a bottleneck. To do this, we analyzed the variability of eighteen microsatellite loci. We found moderately high genetic heterozygosity for wolves from Bosnia and Herzegovina, as described for other Dinaric-Balkan wolf populations. We reveal weak genetic structuring with two genetic clusters identified. Wolves from the eastern part of the region formed a relatively distinct cluster, whereas individuals in the second cluster overlapped quite considerably with admixed individuals. Despite the signal of genetic structure being weak, clustering of individuals from the eastern part of the country extended through all analyses. Thus, this cluster could be considered a separate management unit, perhaps requiring specific conservation attention.
... The divergent profiles of Ukrainian Steppe wolves may to some extent be a result of immigration from outside the study area. The Ukrainian part of our study area could be receiving immigrants from the steppe or foreststeppe regions farther east and north, and similar immigration from eastern and northern regions may occur in the western Russian part of our study area (Pilot 2005). F ST values for 67-K loci were lowest between northcentral Europe and Ukrainian Steppe wolves . ...
Ecological and environmental heterogeneity can produce genetic differentiation in highly mobile species. Accordingly, local adaptation may be expected across comparatively short distances in the presence of marked environmental gradients. Within the European continent, wolves (Canis lupus) exhibit distinct north–south population differentiation. We investigated more than 67-K single nucleotide polymorphism (SNP) loci for signatures of local adaptation in 59 unrelated wolves from four previously identified population clusters (northcen-tral Europe n = 32, Carpathian Mountains n = 7, Dinaric-Balkan n = 9, Ukrai-nian Steppe n = 11). Our analyses combined identification of outlier loci with findings from genome-wide association study of individual genomic profiles and 12 environmental variables. We identified 353 candidate SNP loci. We examined the SNP position and neighboring megabase (1 Mb, one million bases) regions in the dog (C. lupus familiaris) genome for genes potentially under selection, including homologue genes in other vertebrates. These regions included functional genes for, for example, temperature regulation that may indicate local adaptation and genes controlling for functions universally important for wolves, including olfaction, hearing, vision, and cognitive functions. We also observed strong outliers not associated with any of the investigated variables, which could suggest selective pressures associated with other unmea-sured environmental variables and/or demographic factors. These patterns are further supported by the examination of spatial distributions of the SNPs associated with universally important traits, which typically show marked differences in allele frequencies among population clusters. Accordingly, parallel selection for features important to all wolves may eclipse local environmental selection and implies long-term separation among population clusters.
Integrating data across studies with traditional microsatellite genetic markers requires careful calibration and represents an obstacle for investigation of wide-ranging species where populations require transboundary management. We used the “yardstick” method to compare results published across Europe since 2002 and new wolf (Canis lupus) genetic profiles from the Carpathian Mountains in Central Europe and the Dinaric Mountains in Southeastern Europe, with the latter as our reference population. We compared each population with Dinaric wolves, considering only shared markers (range 4–17). For each population, we calculated standard genetic diversity indices plus calibrated heterozygosity (Hec) and allelic richness (Ac). Hec and Ac in Dinaric (0.704 and 9.394) and Carpathian wolves (0.695 and 7.023) were comparable to those observed in other large and mid-sized European populations, but smaller than those of northeastern Europe. Major discrepancies in marker choices among some studies made comparisons more difficult. However, the yardstick method, including the new measures of Hec and Ac, provided a direct comparison of genetic diversity values among wolf populations and an intuitive interpretation of the results. The yardstick method thus permitted the integration of diverse sources of publicly available microsatellite data for spatiotemporal genetic monitoring of evolutionary potential.
In recent years, the golden jackal (Canis aureus) has rapidly expanded its distribution throughout Europe. In Bosnia and
Herzegovina, the jackal has also been found to have increased its range, while establishing a core population in the northern
lowlands of the country. This core population has been augmented by individuals from Serbia and Croatia, indicating an
ongoing gene flow between the neighboring regions. Given the lack of comprehensive population-genetic data in the country,
we aimed to produce the first information on jackal genetic diversity and population structure. We analyzed 24 autosomal
microsatellite loci and found moderately high genetic diversity (Na = 5.83, Ho = 0.56, He = 0.59) in 48 jackals from Bosnia
and Herzegovina, comparable to most other European populations. Analyses of population structure revealed two genetic
clusters (K = 2) with an uncertain spatial pattern. Given the continuing expansion of its populations throughout Europe, the
distribution of golden jackal is gradually overlapping with that of humans and other Canidae. Consequently, conflicts are
steadily increasing, as are additional concerns regarding potential hybridization with dogs and wolves. This study is therefore
a valuable contribution to the understanding of future actions and measures that should be established for jackals in Bosnia
and Herzegovina.
The grey wolf (Canis lupus) is one of the most challenging species to conserve in our modern and crowded world. Due to various factors, most European wolf populations are currently growing. In Hungary, numbers have increased since the 2000s. Although spontaneous recolonisation from Slovakia is considered to be the most likely mechanism by the majority of experts, some stakeholders claim that hand-reared individuals have been released. To determine the origin of wolves in northern Hungary, we analysed samples of free-ranging wolves collected in Slovakia and Hungary as well as samples from wolves in private enclosures in the region. We also included reference samples from domestic dogs. All samples were genotyped at 14 canine autosomal tetranucleotide microsatellite loci (STR) and analysed using multivariate, Bayesian methods. Hungarian wolf samples were also analysed using kinship methods. In the free-ranging wolf samples, all loci were polymorphic with 3–12 alleles. The overall observed (Ho) and unbiased expected (uHE) heterozygosities were 0.60–0.66 and 0.69–0.71, respectively. Parental and sibling relationships were also found among Hungarian individuals: three generations of a pack in the Bükk Mountains were identified. Samples from free-ranging wolves clustered separately from those of captive wolves and dogs. However, genetic similarities were found between Slovakian and Hungarian wolf samples. Our analyses indicate a Slovakian origin of the sampled Hungarian wolves, and we found no evidence that individuals originating in captivity have played any role in the recolonisation process. Kinship relationships and moderate genetic diversity suggest that there is ongoing gene flow across the Slovakian–Hungarian border.
Genetic diversity and population structure of the grey wolf (Canis lupus Linnaeus, 1758) and evidence of wolf × dog hybridisation in the centre of European Russia • Miroslav P. Korablev, • Nikolay P. Korablev & • Pavel N. Korablev Mammalian Biology (2020)Cite this article • Metrics Abstract Throughout history the human-wolf interaction has not evolved in favour of the wolf, however, wolves have never been endangered in Russia. The wolf (Canis lupus lupus L., 1758) population in the central part of European Russia is relatively high, where environmental conditions, such as relatively undisturbed habitats, wide forested areas and abundance of natural prey, have always contributed to the long-term survival of the species. The human persecution of wolves has resulted in almost total extinction of the species in many European countries. In Russia, extermination campaigns have led to severe fluctuations in the number of wolves during the second half of the twentieth century, however, since the early 1990s there has been a tendency towards constant growth in the numbers. Previous studies provided preliminary data on population genetics of the wolf population in European Russia and have generally shown homogeneity of the population structure as well as detecting genetic bottleneck. However, the comprehensive study of genetic diversity and population structure during the period following the last severe decline is of great interest. Another important aspect in the study of wolf populations is the assessment of the magnitude of wolf × dog hybridisation, which is a phenomenon of conservation and social significance. We used 101 samples from the wolf population and 32 dogs to examine population structure, genetic diversity and events of interspecific hybridisation in the centre of European Russia, based on analysis of 11 autosomal microsatellites. In the studied region, wolves exhibit a high level of genetic diversity (HE = 0.79 ± 0.03, HO = 0.74 ± 0.01, NA = 10.00 ± 5.02) which
