Genetic characterization of grey wolves (<Emphasis Type="Italic">Canis lupus</Emphasis> L. <Emphasis Type="Italic">1758</Emphasis>) from Bosnia and Herzegovina: implications for conservation

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.     

DNA diversity in the studies of genetic distance among isolated human populations in bosnia

Different degrees of isolation found in various part of Bosnia and Herzegovina may be induced by various factors. Bjelasnica-Treskavica region, located around 40 kilometers southwest from Sarajevo — capital of Bosnia and Herzegovina, is highly specific in that way. We chose three isolated communities: Dejcici, Bobovica and Lukomir for the study of genetic structure of isolated human populations. Based on general data three relative degrees of isolation/openness among the villages have been presumed as follows: first (lower-Dejcici), second (middle — Bobovica) and third (higher — Lukomir) 15 short tandem repeat (STR) loci and hypervariable region of mtDNA were chosen as a markers for study of population structure. Microsatellite allele frequencies, and mtDNA molecular diversity of Heterozigosity and coefficient of gene differentiation across all observed STR loci were estimated. Also, gene and nucleotide diversity of observed mtDNA regions were obtained. Genetic distance between three populations was calculated using method of Reynolds et al. (1983). For analysis of interpopulation relationship based on polymorphism of HV I and HV II region, estimation of pairwise differences was used. results of this research showed consistence with initial hypothesis on divergence based on socio-cultural factors.     

Effects of forced migrations on spatial and temporal changes in genetic structure of two local populations in Bosnia

Ten qualitative traits were observed in two isolated human populations in Bosnia for prewar and postwar periods. Due to recent war in Bosnia and Herzegovina two million citizens was forced to migrate. Dramatic migration effects occurred in the investigated populations. Population characteristics found were geographic isolation, high levels of marital distance “zero”, patrilocality and propagation isolation. Relative recessive allele frequencies of ten qualitative traits were calculated. Average heterozigosity, genetic differentiation, total heterozigosity, within populations heterozigosity, genetic distance and relative measures based on results of genetic distance analysis were observed. The total number of observed individuals was 1875. We have observed 1209 individuals in prewar period (Bijela population 731 and Memici population 478) and 666 individuals in the postwar period (Bijela population 248 and Memici population 478). Results ofFst andGst analysis showed changed degree of genetic differentiation across observed loci. Significant changes in differentiation were recorded for crooked little finger and PTC tasting. Modest changes of gene differentiation were recorded for ear lobe type and thumb proximal extensibility. Genetic distance analisis between Bijela and Memici has lower value after then before the war, but genetic distance between Bijela-prewar and Bijela-postwar has higher value, as well as Memici-prewar and Memici-postwar genetic distance. Genetic distance between Bijela-prewar and Memiciprewar, as well as Bijela-postwar and Memici-postwar showed similarity. Constructed dendrogram based on results of genetic distance analyses indicated two clusters groups (1. Bijela-prewar, Memici-prewar; 2. Bijela-postwar, Memici-postwar). Changed genetic differentiation and results of genetic distance analyses indicated possibility of significant impact of forced migration in genetic structure of observed populations.     

Determination of combined sibship indices “gray zone” using 15 STR loci for central Bosnian human population

In our previous population studies of Bosnia and Herzegovina human population, we have used autosomal STR, Y-STR, and X-STR loci, as well as Y-chromosome NRY biallelic markers. All obtained results were included in Bosnian referent database. In order of future development of applied population molecular genetics researches of Bosnia and Herzegovina human population, we have examined the effectiveness of 15 STR loci system in determination of sibship by using 15 STR loci and calculating different cut-off points of combined sibship indices (CSI) and distribution of sharing alleles. From the perspective of its application, it is very difficult and complicated to establish strict CSI cut-off values for determination of the doubtless sibship. High statistically significant difference between the means of CSI values and in distribution of alleles sharing in siblings and non-siblings was noticed (P < 0.0001). After constructing the “gray zone”, only one false positive result was found in three CSI cut-off levels with the highest percent of determined sibship/non-sibship at the CSI = 0.067, confirming its practical benefit. Concerning the distribution of sharing alleles, it is recommended as an informative estimator for its usage within Bosnia and Herzegovina human population.     

Wolf population genetics in Europe: a systematic review,meta‐analysis and suggestions for conservation and management

The grey wolf (Canis lupus) is an iconic large carnivore that has increasingly been recognized as an apex predator with intrinsic value and a keystone species. However, wolves have also long represented a primary source of human–carnivore conflict, which has led to long‐term persecution of wolves, resulting in a significant decrease in their numbers, genetic diversity and gene flow between populations. For more effective protection and management of wolf populations in Europe, robust scientific evidence is crucial. This review serves as an analytical summary of the main findings from wolf population genetic studies in Europe, covering major studies from the ‘pre‐genomic era’ and the first insights of the ‘genomics era’. We analyse, summarize and discuss findings derived from analyses of three compartments of the mammalian genome with different inheritance modes: maternal (mitochondrial DNA), paternal (Y chromosome) and biparental [autosomal microsatellites and single nucleotide polymorphisms (SNPs)]. To describe large‐scale trends and patterns of genetic variation in European wolf populations, we conducted a meta‐analysis based on the results of previous microsatellite studies and also included new data, covering all 19 European countries for which wolf genetic information is available: Norway, Sweden, Finland, Estonia, Latvia, Lithuania, Poland, Czech Republic, Slovakia, Germany, Belarus, Russia, Italy, Croatia, Bulgaria, Bosnia and Herzegovina, Greece, Spain and Portugal. We compared different indices of genetic diversity in wolf populations and found a significant spatial trend in heterozygosity across Europe from south‐west (lowest genetic diversity) to north‐east (highest). The range of spatial autocorrelation calculated on the basis of three characteristics of genetic diversity was 650?850 km, suggesting that the genetic diversity of a given wolf population can be influenced by populations up to 850 km away. As an important outcome of this synthesis, we discuss the most pressing issues threatening wolf populations in Europe, highlight important gaps in current knowledge, suggest solutions to overcome these limitations, and provide recommendations for science‐based wolf conservation and management at regional and Europe‐wide scales.     

The Genetics of Bene Israel from India Reveals Both Substantial Jewish and Indian Ancestry

The Bene Israel Jewish community from West India is a unique population whose history before the 18^th century remains largely unknown. Bene Israel members consider themselves as descendants of Jews, yet the identity of Jewish ancestors and their arrival time to India are unknown, with speculations on arrival time varying between the 8th century BCE and the 6th century CE. Here, we characterize the genetic history of Bene Israel by collecting and genotyping 18 Bene Israel individuals. Combining with 486 individuals from 41 other Jewish, Indian and Pakistani populations, and additional individuals from worldwide populations, we conducted comprehensive genome-wide analyses based on F_ST, principal component analysis, ADMIXTURE, identity-by-descent sharing, admixture linkage disequilibrium decay, haplotype sharing and allele sharing autocorrelation decay, as well as contrasted patterns between the X chromosome and the autosomes. The genetics of Bene Israel individuals resemble local Indian populations, while at the same time constituting a clearly separated and unique population in India. They are unique among Indian and Pakistani populations we analyzed in sharing considerable genetic ancestry with other Jewish populations. Putting together the results from all analyses point to Bene Israel being an admixed population with both Jewish and Indian ancestry, with the genetic contribution of each of these ancestral populations being substantial. The admixture took place in the last millennium, about 19–33 generations ago. It involved Middle-Eastern Jews and was sex-biased, with more male Jewish and local female contribution. It was followed by a population bottleneck and high endogamy, which can lead to increased prevalence of recessive diseases in this population. This study provides an example of how genetic analysis advances our knowledge of human history in cases where other disciplines lack the relevant data to do so.     

Validity and reliability of the Oral Impacts on Daily Performance (OIDP) scale in the elderly population of Bosnia and Herzegovina

doi: 10.1111/j.1741‐2358.2011.00584.x Validity and reliability of the Oral Impacts on Daily Performance (OIDP) scale in the elderly population of Bosnia and Herzegovina Objectives: To adapt the Oral Impacts on Daily Performance (OIDP) index for elderly people in Bosnia and Herzegovina and test its validity, reliability and responsiveness to change. Background: Clinical measures alone may not be adequate for assessing the oral health of individuals. Subjective oral health indicators tested within a particular cultural context may not be relevant across cultures. Materials and methods: The study population comprised 231 free‐living adults aged 65 years or older. The OIDP was cross‐culturally adapted from English into the Serbian language and its psychometric properties were tested. Data were collected using a clinical examination and a questionnaire containing the OIDP. Results: In terms of reliability, Cronbach’s alpha coefficient was 0.82 and the intraclass correlation coefficient 0.88. The very high correlation of OIDP with self‐rated oral health (r = 0.78) verified criterion validity, while construct validity was demonstrated through its significant and graded associations with other subjective health measures. OIDP change scores on a treated subsample showed moderate effect size (0.59) and were associated with perceptions of oral health change, providing evidence for its responsiveness to change. Conclusion: The Bosnian version of the OIDP showed satisfactory validity, reliability and responsiveness to change confirming its appropriateness for use among older populations in Bosnia and Herzegovina.     

Microsatellite markers confirm extensive population fragmentation of the endangered Balkan palaeoendemic Martino’s vole (<Emphasis Type="Italic">Dinaromys bogdanovi</Emphasis>)

The Martino’s vole is an endangered rodent endemic to the western Balkan Peninsula. Its range is fragmented, and populations are small due to high habitat specificity. The level of genetic variation within such populations is often low, and genetic differentiation between patchily dispersed populations is high. By scoring eight microsatellite loci in 110 individual Martino’s voles originating from 27 locations throughout the species range, we analysed genetic variation at both the intra- and interpopulation level. Factorial correspondence analysis, Bayesian analyses, and allele sharing distances divided individuals into three phylogroups (Northwestern, Central, and Southeastern), thus providing independent support for phylogeographic structuring, a pattern that has been described in previous studies based on mitochondrial DNA. Spatial genetic analyses showed that populations are highly fragmented, even in those areas with the highest population densities. The highest intrapopulation genetic variability and stable effective population sizes were found in Mount Zelengora (Bosnia and Herzegovina), which harbours a relatively large population of Martino’s voles. Populations in the Central and Southeastern lineages exhibited a significant isolation-by-distance pattern, indicating limited gene flow between them. Contrary to previous opinion, low effective population size and very limited gene flow between remaining populations suggest that the long-term existence of the Martino’s vole might not be secure, even in populations that live in optimal habitats. The only threat to the Martino’s vole identified thus far is competitive exclusion by the European snow vole. However, our results suggest that conservation problems associated with this paleoendemic rodent are more complex.     

Genetic diversity and structuring of the grey wolf population from the Central Balkans based on mitochondrial DNA variation

The Dinaric-Balkan grey wolf population used to be at a border between the large remaining Eastern European populations and the largely eradicated Western European populations. During the last few decades we have witnessed the Western European wolf population recovery. Substantial genetic variation has previously been reported in the Balkan wolf population, but rigorous genetic characterization has not been done for its central parts. The aims of this research were to determine genetic diversity based on mtDNA sequence variability, to infer possible population structuring, to find genetic signals of population expansions or bottlenecks and to evaluate phylogenetic position of the grey wolf population from the Central Balkans. Six haplotypes were detected, of which three have only been found in the Balkan region. These haplotypes belong to both haplogroups previously determined in Europe. Based on our mtDNA sequence analyses, the Dinaric-Balkan wolf population is vertically differentiated into “western” (Croatia/Bosnia and Herzegovina) and “eastern” (Serbia/Macedonia) subpopulations. None of the results support assumption of population expansion. Instead, significantly positive values for Tajima's D and Fu's Fs may suggest recent population bottleneck. Obtained data may be helpful in observation to which extent gene pool from the Balkans contribute to newly founded populations in Western Europe.     

Does Lilium bosniacum merit species rank? A classical and molecular-cytogenetic analysis

Four populations of the Balkan endemic taxon Lilium bosniacum from Bosnia and Herzegovina were investigated. Conventional karyological study did not reveal any important differences. Molecular-cytogenetic studies pointed out certain interpopulation and intrapopulation variability, and important differences in organization of ribosomal genes in regard to its closest relative L. carniolicum. The results of fluorochrome banding and FISH experiment for L. bosniacum were reported here for the first time. Differences occurred in the number and position of 18S-5.8S-26S ribosomal gene loci for some individuals from population Kladanj. Heterochromatic bands revealed with DAPI after FISH experiment were constant. All investigated populations possess the same number of active NORs except some individuals from Kladanj population. Genome size and GC-bases percentage, estimated by flow cytometry, did not show any significant difference among the populations. However, the present results reveal clear interspecific differentiation between two endemics, L. carniolicum and L. bosniacum.     

A case of acromegaly in the Middle Ages

A case of acromegaly in a male individual aged about 30 years, who lived in the area of Central Balkans in the 14th or 15th century will be described here. This individual is from an archeological site named Przine near Gacko, Bosnia and Herzegovina. The diagnosis is based on pathological changes of the skull and the postcranial skeleton. By radiological analysis it could be observed that the Sella turcica does not show any change. Furthermore, the individual under study is not conspicuous by giant growth. This case of acromegaly is the oldest one found in the Balkan area.     

New distribution data and population structure of the European mudminnow Umbra krameri in Serbia and Bosnia and Herzegovina

Seventy‐six individuals of the European mudminnow Umbra krameri from two recent populations from Serbia (Bakreni Batar and Lugomir) and one from Bosnia and Herzegovina (Gromi?elj) were analysed for habitat preferences and population structure. The population from Lugomir is a newly recorded population in Serbia. Besides this new record, it is noteworthy that all three studied locations are outside the currently known species distribution range limits.     

Leaf anatomy of Carex humilis does not correlate with orographic,geological and bioclimatic habitat conditions in C&SE Europe

This paper presents the results of a multivariate morphometric study of leaf anatomical characters in different, geographically very distant populations of taxon Carex humilis from Hungary, Romania, Serbia, Bosnia & Herzegovina, Montenegro and Albania with the aim to reveal the trends of population differentiation. Analyses were performed on the cross-section of 173 leaves collected from 12 populations. In order to establish the overall morphological variation and relationships between individuals from all populations, principal component analyses (PCA) and canonical discriminant analysis (CDA) have been done. The UPGMA clustering analyses based on leaf anatomical characters and habitat climatic characteristics were carried out to explore whether the observed anatomical differences are a result of adaptive responses. Regression analysis (linear regression) was performed to identify the level of correlation between leaf anatomical characters and basic orographic, geological, and bioclimatic habitat characteristics. Quite unexpectedly, most of observed groups are formed of geographically very distant populations which are living in extremely different climatic and geological conditions, indicating that general anatomical differentiation in Carex humilis in C&SE Europe cannot be explained by the environmental impacts, and basically do not represent an adaptive response to different climatic or geological condition.     

Genetic diversity and differentiation of 12 eastern Adriatic and western Dinaric native sheep breeds using microsatellites

Nuclear genetic diversity and differentiation of 341 sheep belonging to 12 sheep breeds from Croatia and Bosnia and Herzegovina were examined. The aim of the study was to provide the understanding of the genetic structure and variability of the analysed pramenka sheep populations, and to give indications for conservation strategies based on the population diversity and structure information. The genetic variation of the sheep populations, examined at the nuclear level using 27 microsatellite loci, revealed considerable levels of genetic diversity, similar to the diversity found in other European indigenous low-production sheep breeds. Population-specific alleles were detected at most loci and in breeds analysed. The observed heterozygosity ranged from 0.643 (in Lika pramenka) to 0.743 (in Vlasic pramenka), and the expected heterozygosity ranged from 0.646 (in Lika pramenka) to 0.756 (in Dalmatian pramenka). Significant inbreeding coefficients were found for half of the populations studied and ranged from 0.040 (Pag island sheep) to 0.091 (Kupres pramenka). Moderate genetic differentiation was found between the studied sheep populations. The total genetic variability observed between different populations was 5.29%, whereas 94.71% of the variation was found within populations. Cres island sheep, Lika pramenka and Istrian sheep were identified as the most distinct populations, which was confirmed by the factorial analysis of correspondence and supported through a bootstrapping adjustment to correct for the difference in the sample sizes. The population structure analysis distinguished 12 clusters for the 12 sheep breeds analysed. However, the cluster differentiation was low for Dalmatian, Vlasic, Stolac and Krk pramenka. This systematic study identified Lika pramenka and Rab island sheep as those with the lowest diversity, whereas Istrian sheep and Pag island sheep had the highest. Conservation actions are proposed for Istrian, Rab and Cres island sheep, Lika and Kupres pramenka because of high estimated coefficients of inbreeding.     

Understanding human DNA sequence variation

Over the past century researchers have identified normal genetic variation and studied that variation in diverse human populations to determine the amounts and distributions of that variation. That information is being used to develop an understanding of the demographic histories of the different populations and the species as a whole, among other studies. With the advent of DNA-based markers in the last quarter century, these studies have accelerated. One of the challenges for the next century is to understand that variation. One component of that understanding will be population genetics. We present here examples of many of the ways these new data can be analyzed from a population perspective using results from our laboratory on multiple individual DNA-based polymorphisms, many clustered in haplotypes, studied in multiple populations representing all major geographic regions of the world. These data support an "out of Africa" hypothesis for human dispersal around the world and begin to refine the understanding of population structures and genetic relationships. We are also developing baseline information against which we can compare findings at different loci to aid in the identification of loci subject, now and in the past, to selection (directional or balancing). We do not yet have a comprehensive understanding of the extensive variation in the human genome, but some of that understanding is coming from population genetics.     

The Paternal Landscape along the Bight of Benin – Testing Regional Representativeness of West-African Population Samples Using Y-Chromosomal Markers

Patterns of genetic variation in human populations across the African continent are still not well studied in comparison with Eurasia and America, despite the high genetic and cultural diversity among African populations. In population and forensic genetic studies a single sample is often used to represent a complete African region. In such a scenario, inappropriate sampling strategies and/or the use of local, isolated populations may bias interpretations and pose questions of representativeness at a macrogeographic-scale. The non-recombining region of the Y-chromosome (NRY) has great potential to reveal the regional representation of a sample due to its powerful phylogeographic information content. An area poorly characterized for Y-chromosomal data is the West-African region along the Bight of Benin, despite its important history in the trans-Atlantic slave trade and its large number of ethnic groups, languages and lifestyles. In this study, Y-chromosomal haplotypes from four Beninese populations were determined and a global meta-analysis with available Y-SNP and Y-STR data from populations along the Bight of Benin and surrounding areas was performed. A thorough methodology was developed allowing comparison of population samples using Y-chromosomal lineage data based on different Y-SNP panels and phylogenies. Geographic proximity turned out to be the best predictor of genetic affinity between populations along the Bight of Benin. Nevertheless, based on Y-chromosomal data from the literature two population samples differed strongly from others from the same or neighbouring areas and are not regionally representative within large-scale studies. Furthermore, the analysis of the HapMap sample YRI of a Yoruban population from South-western Nigeria based on Y-SNPs and Y-STR data showed for the first time its regional representativeness, a result which is important for standard population and forensic genetic applications using the YRI sample. Therefore, the uniquely and powerful geographical information carried by the Y-chromosome makes it an important locus to test the representativeness of a certain sample even in the genomic era, especially in poorly investigated areas like Africa.     

Population genetic structure of wolves in the northwestern Dinaric‐Balkan region

The Balkan Peninsula and the Dinaric Mountains possess extraordinary biodiversity and support one of the largest and most diverse wolf (Canis lupus) populations in Europe. Results obtained with diverse genetic markers show west‐east substructure, also seen in various other species, despite the absence of obvious barriers to movement. However, the spatial extent of the genetic clusters remains unresolved, and our aim was to combine fine‐scale sampling with population and spatial genetic analyses to improve resolution of wolf genetic clusters. We analyzed 16 autosomal microsatellites from 255 wolves sampled in Slovenia, Croatia, Bosnia and Herzegovina (BIH), and Serbia and documented three genetic clusters. These comprised (1) Slovenia and the regions of Gorski kotar and Lika in Croatia, (2) the region of Dalmatia in southern Croatia and BIH, and (3) Serbia. When we mapped the clusters geographically, we observed west‐east genetic structure across the study area, together with some specific structure in BIH–Dalmatia. We observed that cluster 1 had a smaller effective population size, consistent with earlier reports of population recovery since the 1980s. Our results provide foundation for future genomic studies that would further resolve the observed west‐east population structure and its evolutionary history in wolves and other taxa in the region and identify focal areas for habitat conservation. They also have immediate importance for conservation planning for the wolves in one of the most important parts of the species’ European range.     

Frequency of the ΔF508 deletion and G551D,R553X and G542X mutations in Yugoslav CF patients

Summary A study was undertaken to find the frequency of the F508 deletion and those of the G551D, R553X and G524X mutations among the mainly Slavic population of Serbia, Bosnia, Herzegovina, and Montenegro and compare the frequencies determined with those in other European populations. The F508 mutation was found to account for about 70% of CF genes in central Jugoslavia, where its frequency is significantly higher than elsewhere in Southern European populations.     

A new cytochrome b phylogroup of the common vole (Microtus arvalis) endemic to the Balkans and its implications for the evolutionary history of the species

The phylogeographic architecture of the common vole, Microtus arvalis, has been well‐studied using mitochondrial DNA and used to test hypotheses relating to glacial refugia. The distribution of the five described cytochrome b (cyt b) lineages in Europe west of Russia has been interpreted as a consequence of postglacial expansion from both southern and central European refugia. A recently proposed competing model suggests that the ‘cradle’ of the M. arvalis lineages is in western central Europe from where they dispersed in different directions after the Last Glacial Maximum. In the present study, we report a new cyt b lineage of the common vole from the Balkans that is not closely related to any other lineage and whose presence might help resolve these issues of glacial refugia. The Balkan phylogroup occurs along the southern distributional border of M. arvalis in central and eastern Bosnia and Herzegovina, Montenegro, and eastern Serbia. Further north and west in Slovenia, Bosnia and Herzegovina, and Serbia, common voles belong to the previously‐described Eastern lineage, whereas both lineages are sympatric in one site in Bosnia and Herzegovina. The Balkan phylogroup most reasonably occupied a glacial refugium already known for various Balkan endemic species, in contrast to the recently proposed model. South‐east Europe is an absolutely crucial area for understanding the postglacial colonization history of small mammals in Europe and the present study adds to the very few previous detailed phylogeographic studies of this region. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 788–796.