Parentage Relationships Research Articles

Normative analysis of the paternity presumption in Article 1073 of the Civil Code of China

The legal system of parentage is a mature rule in traditional civil law, encompassing the acknowledgment and denial of parent-child relationships. In China, the parentage legal system outlined in the Civil Code is a guiding provision that requires supplementary operational rules. The rule of parentage presumption, as a crucial component of the parentage legal system, serves as the premise and foundation of the entire parentage system. China’s parentage presumption involves both legal and factual presumptions. To elucidate the legislative intent and objective expression of the legal provisions concerning parentage in the Civil Code, it is necessary to clarify the legal attributes, logic, and effects of parentage presumption. Based on this, deviations in legal application due to inconsistent interpretations by different judges should be identified and adjusted. There exist four challenges in the practical application of the parentage presumption rule within the Chinese judicial system. It is imperative for judges to possess a precise and consistent understanding of these challenges. Article 1073 of the Civil Code and judicial interpretations constitute a remedial mechanism for parentage presumption. However, for this article to yield the intended legal effects, detailed specifications are needed regarding the parties eligible to file denial lawsuits of legitimate children, the grounds for denial, and the statute of limitations. Additionally, the implied recognition system reflected in the legal provision should also be supplemented with unified and applicable regulations to ensure operability within China’s legal system. Only with consistent legislative interpretations and applications can the consistency of legal application in parentage relationships be safeguarded, thereby upholding the authority of the Civil Code.

Performance of SNP markers for parentage analysis in the Italian Alpine brown bear using non-invasive samples

Determination of parentage provides valuable information for the conservation of wild populations, for instance, by allowing the monitoring of breeding success and inbreeding. Between 1999 and 2002, nine brown bears (Ursus arctos) were translocated to augment the remnant population of a few surviving individuals in the Italian Alps, but only part of them reproduced, with a higher inbreeding risk occurrence in the long-time. Currently, in the Alpine population, parentage tests are assessed through the analysis of 15 microsatellite loci (STRs), but the reduction of genetic variability in future generations will need the use of additional informative markers. Single nucleotide polymorphisms (SNPs) have been proven to be useful and reliable in individual identification and family reconstruction; moreover, they can perform well on low-quality samples. In this study, we analysed 51 SNPs to generate a SNP multilocus genotype dataset of 54 Alpine brown bears (Ursus arctos) and compared its performance in parentage analysis with the validated STR dataset. We found that SNPs alone are not sufficient to determine parentage relationships, but the combination of SNPs and STRs provided unambiguous parentage assignments. The combined panel also performed better than STRs when true parents were not present in the dataset and, consequently, showed higher values of assignment probabilities.

Shared pedigree relationships and transmission of unreduced gametes in cultivated banana.

Cultivated bananas resulted from inter(sub)specific hybridizations involving Musa species and subspecies (M. acuminata subspecies, M. schizocarpa, M. balbisiana) and the subsequent selection, centuries ago, of hybrids with parthenocarpic, seedless fruits. Cultivars have low fertility and are vegetatively propagated, forming groups of somaclones. Relatively few of them, mainly triploids, are grown on a large scale and characterization of their parental relationships may be useful for breeding strategies. Here we investigate parental relationships and gamete-type contributions among diploid and polyploid banana cultivars. We used SNP genotyping data from whole-genome sequencing of 178 banana individuals, including 111 cultivars, 55 wild bananas and 12 synthetic F1 hybrids. We analysed the proportion of SNP sites in accordance with direct parentage with a global statistic and along chromosomes for selected individuals. We characterized parentage relationships for 7 diploid cultivars, 11 triploid cultivars and 1 tetraploid cultivar. Results showed that both diploid and triploid cultivars could have contributed gametes to other banana cultivars. Diploids may have contributed 1x or 2x gametes and triploids 1x to 3x gametes. The Mchare diploid cultivar group, nowadays only found in East Africa, was found as parent of two diploid and eight triploid cultivars. In five of its identified triploid offspring, corresponding to main export or locally popular dessert bananas, Mchare contributed a 2x gamete with full genome restitution without recombination. Analyses of remaining haplotypes in these Mchare offspring suggested ancestral pedigree relationships between different interspecific banana cultivars. The current cultivated banana resulted from different pathways of formation, with implication of recombined or un-recombined unreduced gametes produced by diploid or triploid cultivars. Identification of dessert banana's parents and the types of gametes they contributed should support the design of breeding strategies.

Characterization of 35 new microsatellite markers for the blacktip reef shark (Carcharhinus melanopterus) and cross-species amplification in eight other shark species.

Shark species are overfished at a global scale, as they are poached for the finning industry or are caught as bycatch. Efficient conservation measures require fine-scale spatial and temporal studies to characterize shark habitat use, infer migratory habits, analyze relatedness, and detect population genetic differentiation. Gathering these types of data is costly and time-consuming, especially when it requires collection of shark tissue samples. Genetic tools, such as microsatellite markers, are the most economical sampling method for collecting genetic data, as they enable the estimation of genetic diversity, population structure and parentage relationships and are thus an efficient way to inform conservation strategies. Here, a set of 45 microsatellite loci was tested on three blacktip reef shark (Carcharhinus melanopterus) populations from three Polynesian islands: Moorea, Morane and Tenararo. The set was composed of 10 previously published microsatellite markers and 35 microsatellite markers that were developed specifically for C. melanopterus as part of the present study. The 35 novel and 10 existing loci were cross-amplified on eight additional shark species (Carcharhinus amblyrhynchos, C. longimanus, C. sorrah, Galeocerdo cuvier, Negaprion acutidens, Prionacea glauca, Rhincodon typus and Sphyrna lewini). These species had an average of 69% of successful amplification, considered if at least 50% of the individual samples being successfully amplified per species and per locus. This novel microsatellite marker set will help address numerous knowledge gaps that remain, concerning genetic stock identification, shark behavior and reproduction via parentage analysis.

Construction of an individual identification panel for horses using insertion and deletion markers.

Individual identification and paternity testing are important for avoiding inbreeding in the management of small populations of wild and domestic animals. In horse racing industries, they are extremely important for identifying and registering individuals and doping control to ensure fair competition. In this study, we constructed an individual identification panel for horses by using insertion and deletion (INDEL) markers. The panel included 39 INDEL markers selected from a whole-genome INDEL database. Genotyping of 89 Thoroughbreds showed polymorphisms with minor allele frequencies (MAFs) of 0.180-0.489 in all markers. The total probability of exclusion for paternity testing, power of discrimination, and probability of identity were 0.9994271269, >0.9999999999, and 0.9999999987, respectively. The panel was applied to 13 trios (sires, dams, and foals), and no contradictions were observed in genetic inheritance among the trios. When this panel was applied to the trios (52 trios) containing false fathers, an average of 7.3 markers excluded parentage relationships. In addition, genomic DNA extracted from the urine of six horses was partially genotyped for 39 markers, and 6-28 markers were successfully genotyped. The newly constructed panel has two advantages: a low marker mutation rate compared with short tandem repeats and a genotyping procedure that is as simple as short tandem repeat typing compared with single nucleotide variant typing. This panel can be applied for individual identification, paternity determination, and urine-sample identification in Thoroughbred horses.

Evaluation of parentage testing using single nucleotide polymorphism markers for draft horses in Japan.

We evaluated the utility of single nucleotide polymorphism (SNP) markers for parentage testing in Breton (BR) and Percheron (PR) horses in Japan using the proposed International Society for Animal Genetics (P-ISAG) 147 SNP panel and 414 autosomal SNPs. Genomic DNA was extracted from 98 horses of two breeds, BR (n = 47) and PR (n = 51), and sequenced using next-generation sequencing. The average minor allele frequencies for the P-ISAG panel for BR and PR were 0.306 and 0.301, respectively. The combined probabilities of exclusion (PEs) given two parents and one offspring: exclude a relationship (PE01) and given one parent and one offspring: exclude their relationship (PE02) were over 0.9999 for both breeds. Using the P-ISAG panel, no exclusion or doubtful cases were identified in 35 valid parent-offspring pairs, suggesting that the P-ISAG panel is helpful for parentage verification in both breeds. In contrast, as 0.18% of falsely accepted parentages were observed in the parentage discovery cases, additional markers such as the combination of the P-ISAG panel and 414 autosomal SNPs (561-SNP set) presented here should be used to identify valid parent-offspring pairs of horses with unknown parentage relationships.

Diverse parentage relationships in paternal mouthbrooding fishes.

While mouthbrooding is not an uncommon parental care strategy in fishes, paternal mouthbrooding only occurs in eight fish families and is little studied. The high cost of paternal mouthbrooding to the male implies a low risk of investment in another male's offspring but genetic parentage patterns are poorly known for paternal mouthbrooders. Here, we used single-nucleotide polymorphism genetic data to investigate parentage relationships of broods of two mouthbrooders of northern Australian rivers, mouth almighty Glossamia aprion and blue catfish Neoarius graeffei. For N. graeffei, we found that the parentage pattern was largely monogamous with the brooder male as the sire. For G. aprion, the parentage pattern was more heterogeneous including observations of monogamous broods with the brooder male as the sire (73%), polygyny (13%), cuckoldry (6%) and a brood genetically unrelated to the brooder male (6%). Findings demonstrate the potential for complex interrelationships of male care, paternity confidence and mating behaviour in mouthbrooding fishes.

Waste not, want not: Microsatellites remain an economical and informative technology for conservation genetics.

Comparisons of microsatellites and single‐nucleotide polymorphisms (SNPs) have found that SNPs outperform microsatellites in population genetic analyses, questioning the continued utility of microsatellites in population and landscape genetics. Yet, highly polymorphic markers may be of value in species that have reduced genetic variation. This study repeated previous analyses that used microsatellites with SNPs developed from ddRAD sequencing in the black‐capped vireo source‐sink system. SNPs provided greater resolution of genetic diversity, population differentiation, and migrant detection but could not reconstruct parentage relationships due to insufficient heterozygosities. The biological inferences made by both sets of markers were similar: asymmetrical gene flow from source sites to the remaining sink sites. With the landscape genetic analyses, we found different results between the two molecular markers, but associations of the top environmental features (riparian, open habitat, agriculture, and human development) with dispersal estimates were shared between marker types. Despite the higher precision of SNPs, we find that microsatellites effectively uncover population processes and patterns and are superior for parentage analyses in this species with reduced genetic diversity. This study illustrates the continued applicability and relevance of microsatellites in population genetic research.

Correcting parentage relationships in the endangered California Condor: Improving mean kinship estimates for conservation management

Abstract Maintaining the existing biodiversity of endangered species is a goal of conservation management programs, and a major component of many collaborative efforts undertaken by zoos, field biologists, and conservation scientists. Over the past 3 decades, the San Diego Zoo has performed long-term genetic studies in support of the recovery program for the critically endangered California Condor (Gymnogyps californianus). This work has included sex determination of hatchlings and parentage confirmation using microsatellite genotyping. This paper describes the genetic work associated with the California Condor recovery program, which aims to provide a highly accurate pedigree for making informed captive pairing and release recommendations. Initial genotyping began after reintroduced California Condors started reproducing, and the focus was on birds hatched from their wild-laid eggs. However, genetic analysis showed discrepancies relative to behavioral observations of wild birds and led to the species-wide testing of all available samples. This genetic study has resolved instances of individual misidentifications and parental misassignments, verified the first cases of extra-pair paternity in this species, identified parentage where chicks were observed being raised by trios, and found 2 apparent de novo mutations in the captive condor population. Correcting the California Condor pedigree according to genetic parental analysis has produced more accurate estimates of mean kinship values among living birds, ranking potential breeders according to their actual breeding value and helping managers to make informed decisions about captive pairing and release of condors in the wild.

Unraveling the genetic origin of \u2018Glera\u2019, \u2018Ribolla Gialla\u2019 and other autochthonous grapevine varieties from Friuli Venezia Giulia (northeastern Italy)

‘Glera’ and ‘Ribolla Gialla’ are the most economically relevant local grapevine cultivars of Friuli Venezia Giulia region (north-eastern Italy). ‘Glera’ is used to produce the world-renowned Prosecco wine. ‘Ribolla Gialla’ cultivation is constantly increasing due to the strong demand for sparkling wine and is the most important variety in Brda (Slovenia). Knowledge of local varieties history in terms of migration and pedigree relationships has scientific and marketing appeal. Following prospections, genotyping and ampelographic characterization of minor germplasm in Friuli Venezia Giulia, a further research was developed to understand the parentage relationships among the grapevine varieties grown in this region. An integrated strategy was followed combining the analysis of nuclear and chloroplast microsatellites with the Vitis 18k SNP chip. Two main recurrent parents were found, which can be regarded as “founders”: ‘Vulpea’, an Austrian variety parent-offspring related with at least ten Friuli Venezia Giulia cultivars, among them ‘Glera’, and ‘Refosco Nostrano’, first degree related with other six Friuli Venezia Giulia varieties. ‘Ribolla Gialla’ was shown to be another member of the impressively long list of offspring derived from the prolific ‘Heunisch Weiss’. Combining molecular markers and historical references was a high-performance strategy for retracing and adjusting the history of cultivars.

In situ estimation of genetic variation of functional and ecological traits in Quercus petraea and Q.robur.

Predicting the evolutionary potential of natural tree populations requires the estimation of heritability and genetic correlations among traits on which selection acts, as differences in evolutionary success between species may rely on differences for these genetic parameters. In situ estimates are expected to be more accurate than measures done under controlled conditions which do not reflect the natural environmental variance. The aim of the current study was to estimate three genetic parameters (i.e. heritability, evolvability and genetic correlations) in a natural mixed oak stand composed of Quercus petraea and Quercus robur about 100 years old, for 58 traits of ecological and functional relevance (growth, reproduction, phenology, physiology, resilience, structure, morphology and defence). First we estimated genetic parameters directly in situ using realized genomic relatedness of adult trees and parentage relationships over two generations to estimate the traits additive variance. Secondly, we benefited from existing ex situ experiments (progeny tests and conservation collection) installed with the same populations, thus allowing comparisons of in situ heritability estimates with more traditional methods. Heritability and evolvability estimates obtained with different methods varied substantially and showed large confidence intervals, however we found that in situ were less precise than ex situ estimates, and assessments over two generations (with deeper relatedness) improved estimates of heritability while large sampling sizes are needed for accurate estimations. At the biological level, heritability values varied moderately across different ecological and functional categories of traits, and genetic correlations among traits were conserved over the two species. We identified limits for using realized genomic relatedness in natural stands to estimate the genetic variance, given the overall low variance of genetic relatedness and the rather low sampling sizes of currently used long term genetic plots in forestry. These limits can be overcome if larger sample sizes are considered, or if the approach is extended over the next generation.

Cultivated Olive Diversification at Local and Regional Scales: Evidence From the Genetic Characterization of French Genetic Resources.

Molecular characterization of crop genetic resources is a powerful approach to elucidate the origin of varieties and facilitate local cultivar management. Here we aimed to decipher the origin and diversification of French local olive germplasm. The 113 olive accessions of the ex situ collection of Porquerolles were characterized with 20 nuclear microsatellites plus their plastid haplotype. We then compared this collection to Mediterranean olive varieties from the Worldwide Olive Germplasm Bank of Marrakech, Morocco. High genetic diversity was observed within local French varieties, indicating a high admixture level, with an almost equal contribution from the three main Mediterranean gene pools. Nearly identical and closely related genotypes were observed among French and Italian/Spanish varieties. A high number of parent–offspring relationships were also detected among French varieties and between French and two Italian varieties (‘Frantoio’ and ‘Moraiolo’) and the Spanish variety (‘Gordal Sevillana’). Our investigations indicated that French olive germplasm resulted from the diffusion of material from multiple origins followed by diversification based on parentage relationships between varieties. We strongly suggest that farmers have been actively selecting olives based on local French varieties. French olive agroecosystems more affected by unexpected frosts than southernmost regions could also be seen as incubators and as a bridge between Italy and Spain that has enhanced varietal olive diversification.

Molecular Genetics Unveiled Unknown Family Relationships and Hybrids in an Ex-Situ Colony of African Penguins (Spheniscus demersus).

Genealogical relationships among colony members, inbreeding status, and presence of hybrids are crucial data that can assist zoo curators in captive colony management and decision-making on relocation for reproduction. This study employed molecular markers to study a large colony (n = 56) of African Penguin hosted in an Italian biopark. A panel of 15 STRs (single tandem repeats) was selected, and genotype data were analyzed using COLONY software to determine parentage relationships and compare the existing studbook information to a pedigree built from genetic analyses. The existence of extra-pair mating and the presence of hybrids were investigated: discrepancies in kinship relationships emerged following molecular parentage analysis and 10 unknown genetic relationships were revealed. Infidelity of one member of the pair was observed in 6 cases and extra-pair copulation was assessed by genetic analysis in 2 episodes. One member of the colony was found to be a hybrid (S. demersus × S. humboldti); his progeny, derived by extra-pair copulation, was traced. Three other hidden hybrids were discovered and assessed using the identified candidate private alleles. Overall, our results demonstrate that molecular methods to confirm parentage and analyze relatedness among colony members are a valuable tool to complement studbook-based genetic management of African penguin captive populations. Because a variety of behavioral dynamics (e.g., extra-pair mating) can make observations ineffective in some species and because molecular markers outperform studbook in identifying the presence of hybrids, reliance on studbook information alone is not recommended.

Identification of subspecies and parentage relationship by means of DNA fingerprinting in two exemplary of Pan troglodytes (Blumenbach, 1775) (Mammalia Hominidae)

Identification of subspecies and parentage relationship by means of DNA fingerprinting in two exemplary of Pan troglodytes (Blumenbach, 1775) (Mammalia Hominidae)

Parentage determination of cuttlefish (Sepiella japonica) based on microsatellite DNA markers

Abstract Microsatellite markers have been used for more than ten years to elucidate parentage relationships in aquaculture species. This study aimed to assess the feasibility of utilizing microsatellite markers for parentage determination in cuttlefish (Sepiella japonica) using simulations and real data analysis. We developed a panel of eight microsatellite markers in our lab. These markers were highly polymorphic with a mean of 10.1 alleles and an average expected heterozygosity value of 0.809. Using five simple sequence repeat markers, an allele frequency data-based simulation indicated that the combined exclusion probability values would be over 99%, whereas the rate of assignment success for the real data set was 91.8%. Mismatches caused by null alleles and scoring errors at microsatellite loci were the major reasons for the discrepancies between the simulations and real data analysis. We concluded that microsatellite markers can be used as a powerful tool to evaluate the effectiveness of enhancement and release programs for S. japonica.

Paternity Analysis of Wood Turtles (Glyptemys insculpta) Reveals Complex Mating Patterns

Mating system characteristics are of great importance as they may influence male and female reproductive success and reproductive isolation. The wood turtle (Glyptemys insculpta) is a terrestrial freshwater species listed as endangered by the International Union for Conservation of Nature. Considering its conservation status and the paucity of information currently available on parentage relationship for the species, we performed a microsatellite analysis to study the mating system of wood turtles in the Shawinigan River (Québec). We sampled 38 clutches over 2 years (14 in 2006 and 24 in 2007), for a total of 248 offspring genotyped with 7 microsatellite loci. The reconstructed genotypes of the fathers revealed that reproductive success in the sampled clutches varied greatly between males and are positively correlated with the number of mates and clutches sired. Frequency of multiple paternity was estimated at 37% through a consensus of 3 different estimation methods. Positive correlation was observed between the genetic diversity of clutches and the number of fathers. Repeat paternity, however, was observed in 88% of the clutches by the same female in successive years, which suggests either a frequent use of sperm storage, or remating with the same partner in successive years.

Evaluating the success of wildlife crossing structures using genetic approaches and an experimental design: Lessons from a gliding mammal

Abstract Millions of dollars are spent on wildlife crossing structures intended to reduce the barrier effects of roads on wildlife. However, we know little about the degree to which these structures facilitate dispersal and gene flow. Our study incorporates two elements that are rarely used in the evaluation of wildlife crossing structures: an experimental design including a before and after comparison, and the use of genetic techniques to demonstrate effects on gene flow at both population and individual levels. We evaluated the effect of wildlife crossing structures (canopy bridges and glider poles) on a gliding mammal, the squirrel glider (Petaurus norfolcensis). We genotyped 399 individuals at eight microsatellite markers to analyse population structure, first‐generation migrants and parentage relationships. We found that the freeway was not a complete genetic barrier, with a strong effect evident at only one site. We hypothesise that the presence of corridors alongside the freeway and throughout the surrounding landscape facilitated circuitous detours for squirrel gliders. Installing a crossing structure at the location with a strong barrier effect restored gene flow within just 5 years of mitigation. Synthesis and applications. Our study highlights the importance of using genetic techniques not just to evaluate the success of road‐crossing structures for wildlife, but also to guide their placement within the landscape. Managers wishing to reduce the effects of linear infrastructure on squirrel gliders and other arboreal mammals should aim to preserve and enhance vegetation along roadsides and within centre medians, as well as mitigate large gaps by implementing wildlife crossing structures.

Significance of mate selection and adult sex ratio in tiger reintroduction/reinforcement programs

AbstractTranslocations of large carnivores like tiger to restore extirpated populations are increasingly becoming important conservation tools in the face of persistent anthropogenic pressures and relentless habitat loss. Reintroduction success in Panna Tiger Reserve, India was variously attributed to high genetic diversity, less anthropogenic disturbance and stress, efficient execution and technological integrations. Here, we establish parentage relationships and explore the potential of mate choice and appropriate adult sex ratio to explain the successful reintroduction in Panna. We used a combination of radio‐telemetry and genetic analyses to understand mating interactions of translocated tigers and their outcomes. Surprisingly, relatedness and parentage analyses of nine cubs born in 2010–2012 suggested that five cubs (55.5%) were not sired by the single translocated male tiger, emphasizing the difficulty of assessing the persistence of tiger presence over a large geographic area. Genotype data of tigers originally from Panna prior to the present reintroduced population revealed that four cubs were sired by an original Panna tiger which possibly ranged transiently within the released tigers’ home ranges. Presence of additional male for mate choice can be a strong factor for successful reintroduction of tigers in Panna. Although number of females released is a good indicator of reintroduction success, the number of males released is also a very important factor for success. We hypothesize that while tigers live in female‐dominated population structure within male territories, mate selection strategies operate strongly between sexes. Translocation programs should therefore deliberate on this aspect of mate selection and accordingly consider founder compositions to achieve desirable results. This also reinforces the need for post‐release monitoring and thorough investigations into intraspecific interactions and behavior which will facilitate proper management and revival of tiger populations.

Renewal of wild grapevine (Vitis vinifera L. subsp. sylvestris (Gmelin) Hegi) populations through sexual pathway: Some Italian case studies

Wild Euro-Asiatic grapevine (Vitis vinifera L. subsp. sylvestris (Gmelin) Hegi) is an opportunistic liana with reproductive patterns characterized both by vegetative propagation and sexual reproduction. Vegetative propagation would ensure a rapid vine regeneration and land colonization, while sexual reproduction would assure genetic recombination and chromosome re-assortment crucial for evolution and survival of the species. The aim of this study was the analysis of the parentage relationships within and between fourteen Italian wild grapevine populations, in order to characterize the natural mating system of this taxon. Beside parentage relationship analyses, the estimation of genetic diversity, genetic distance and genetic differentiation were assessed. The populations showed a good extent of genetic diversity and low levels of genetic distance and isolation between them, indicating gene flow among sites. This result was supported by parentage relationships between different populations. Nevertheless, populations from North Italy showed the lowest genetic diversity, while internal relatedness and inbreeding estimation values were higher than the populations from Central and Southern Italy. One of the northern populations resulted particularly high in inbreeding intensity. The parentage relationship analysis pointed out in all sites a, sometimes active, sexual reproduction within populations. Seed dispersal by frugivorous vectors among neighbouring sites seemed less frequent and depending on specific environmental circumstances.

Genetic clustering and parentage analysis of Western Balkan grapevines (Vitis vinifera L.)

A total of 90 grapevine samples collected in five countries of the Western Balkan region were evaluated for trueness-to-type and kinship relations based on comparative analysis with 1,130 grapevine genotypes held at the INRA "Domaine de Vassal" French Grape Germplasm Repository, using 14 microsatellite markers. In the context of the comparative analysis, twenty-four synonyms/counterparts and the putative parents for twelve Balkan accessions were identified. We discovered five pairs of homonyms, subsequently confirming the identity or parentage of three of them. Some of the examined accessions were identified either on the basis of the genotypes found in the literature, or through parentage relationships revealed in this study. For the remaining fifty accessions we were unable to establish either their pedigree or to identify them on the basis of SSR profiles available elsewhere. Finally, the Balkan genotypes that were not well classified by synonymy or parentage analysis were further studied with a Principal Coordinate Analysis to reveal genetic clustering within larger datasets of genotypes. The graphical display of the individual and group distances showed that about forty accessions (85 %) are structured within a group of Balkan and Eastern Europe genotypes and only a minor proportion resulted in admixed population assignment.