Kuri cattle of the Lake Chad Basin represent a geographically restricted African cattle population characterised by ecological adaptation and a complex taurine–zebu genomic background. The present study aimed to characterise variation in selected functional nuclear genes together with mitochondrial DNA diversity in eight Kuri cattle individuals (n = 8), including six bulls and two cows. A fragment of the mitochondrial D-loop region was sequenced to assess maternal lineage diversity, while nuclear loci associated with milk protein composition (CSN2, CSN3, CSN1S1, CSN1S2, LGB), lipid metabolism (DGAT1), and thermoregulation (PRLR/SLICK locus) were analysed using PCR and Sanger sequencing. Mitochondrial analysis revealed the presence of multiple maternal haplotypes within the examined sample. Nuclear genotyping demonstrated polymorphism at several milk protein loci, including the β-casein gene (CSN2), where both A1 and A2 alleles were detected. The DGAT1 locus also showed allelic variation, whereas the analysed SLICK-associated region was monomorphic in all individuals. Although based on a limited number of animals, the results indicate the persistence of functional genetic variability in this locally adapted cattle population and provide baseline molecular data for future conservation and breeding studies.

The polymorphism of the bovine β-casein gene (CSN2) is of increasing interest due to its relevance for A2 milk production. This study genotyped 2773 Holstein-Friesian cows for five CSN2 alleles (A1, A2, A3, B, I) using both conventional DNA-based PCR and a newly evaluated direct PCR protocol. Eleven genotypes were detected, with A2/A2 (33.9%) and A1/A2 (30.3%) being the most common, resulting in an A2 allele frequency of 59.0%. Genetic diversity indices indicated moderate polymorphism and a significant deviation from Hardy–Weinberg equilibrium, consistent with ongoing selection for the A2 allele. Associations between CSN2 genotype and milk traits (305-day milk, fat, and protein yield; fat% and protein%) were evaluated using linear mixed-effects models including lactation number, age at calving, and calving year as covariates, and cow ID as a random intercept. Several genotype effects reached statistical significance (p < 0.05); however, all effect sizes were very small (partial η2 < 0.01), indicating that any influence of CSN2 on production traits is negligible within this population and management context. These findings suggest that A2-oriented selection is unlikely to compromise productivity. The direct PCR genotyping method achieved 96–100% success and enabled substantially faster and more cost-efficient processing (approximately 80–90% reduction in reagent costs), providing a rapid and scalable approach for large herds.

Background/Objectives: African swine fever (ASF) has caused substantial wild boar (Sus scrofa) mortality across Europe and prompted intensive density-reduction measures, yet its long-term effects on population genetics remain poorly understood. To our knowledge, this is the first study to compare genetic diversity, population structure, demographic signatures, and wildlife forensic identification parameters in the same wild boar population before ASF emergence and after several years of ASF-related management. Methods: Wild boar samples collected before ASF emergence in Hungary (n = 67) were compared with samples collected after several years of ASF-related management from the same geographic area (n = 65). All 132 individuals were genotyped at 13 tetrameric microsatellite loci. Diversity was assessed using allelic richness, allele number, and heterozygosity; differentiation using F-statistics, analysis of molecular variance, and discriminant analysis of principal components; demographic history using a two-phase mutation model; and forensic performance using probability of identity and probability of identity among siblings. Results: The post-ASF population showed lower genetic diversity and significant temporal differentiation. Paired Wilcoxon tests indicated a significant reduction in the number of alleles, whereas changes in heterozygosity were not significant. Bottleneck analyses provided no robust evidence of a recent genetic bottleneck. Forensic discrimination power declined modestly while remaining sufficient for individual identification. Conclusions: Because no contemporaneous unaffected reference population was available, these changes cannot be attributed specifically to ASF or management rather than to genetic drift or natural turnover, although they are consistent with sustained demographic disturbance. Nevertheless, this study provides the first temporal genetic comparison of a European wild boar population before ASF emergence and after several years of ASF-related management and demonstrates that a routinely used STR marker panel remains suitable for wildlife forensic individual identification despite these genetic changes.

Targeted CSN2 codon-67 genotyping supports A2-oriented breeding and herd management. We compared Kompetitive Allele-Specific PCR (KASP) with a locally implemented gel-based tetra-primer amplification refractory mutation system PCR (T-ARMS PCR) workflow in a balanced, non-population-representative Holstein–Friesian panel using re-audited Sanger-supported reference classifications. Both assays distinguish His67-associated (A1-type) from Pro67-associated (A2-type) β-casein classes rather than complete CSN2 alleles. The paired analysis included 96 independent samples. KASP produced 90 reference-concordant, four reference-discordant, and two no-call outcomes (93.8% all-record reference-concordant yield), whereas T-ARMS PCR produced 77 reference-concordant, six reference-discordant, and 13 ambiguous outcomes (80.2%). Concordance among callable/evaluable results was 95.7% and 92.8%, respectively. In the primary comparison, KASP showed a 13.5 percentage point higher yield (95% CI, 4.2–22.9 percentage points; p = 0.015), mainly because it generated fewer non-interpretable outcomes. All callable KASP discordances involved Pro67/Pro67 reference samples called His67/Pro67. Because no independent DNA dilution, new DNA extraction, repeat KASP run, or new Sanger sequencing was performed, their cause and reproducibility remain unresolved. Formal repeatability and between-run reproducibility were not systematically evaluated; therefore, this comparison is not a formal assay validation and does not define T-ARMS PCR performance beyond the tested local configuration. Discordant, ambiguous, or high-impact classifications require independent repeat testing or sequencing confirmation.

Pathological and microbiological surveillance of wildlife can reveal clinically silent but biologically important reproductive disorders. This case report describes a middle-aged (6–7 years) fallow deer hind (Dama dama) in good body condition, legally culled in Hungary, in which post-mortem examination identified a uterine leiomyoma in the left uterine horn and retained fetal cranial bones in the cranial vagina. To the best of our knowledge, this is the first published description of uterine leiomyoma in this species. Gross pathology, histopathology, and desmin immunohistochemistry supported the diagnosis of leiomyoma, and 16S rRNA amplicon sequencing was used to compare the microbiomes of the unaffected uterine horn, affected uterine tissue, and feces. The affected uterine sample showed a microbial profile more similar to that of feces than the unaffected uterine sample, with an increased relative abundance of genera, including Bacteroides, Escherichia–Shigella, and Turicibacter. As this was a single post-mortem case, no treatment was administered. These findings suggest a possible association between chronic mechanical obstruction, retained fetal material, and marked local microbial alteration, while also illustrating the limitations of causal inference from single-animal microbiome data. This case expands the differential diagnosis of reproductive tract lesions in wild ruminants and highlights the value of integrating pathology with careful microbiome interpretation in wildlife case reports.

This study presents a structured narrative review integrating methodological and decision-oriented perspectives. Milk proteins, particularly β-casein, have attracted increasing scientific and commercial attention due to their genetic variability and role in dairy production and product differentiation. Among β-casein variants, the A1 and A2 alleles of the CSN2 gene are of particular relevance, as their single-nucleotide difference has influenced breeding strategies and the expansion of A2-oriented dairy markets. Although multiple validated molecular genotyping approaches are available for CSN2 A1/A2 discrimination, guidance on their context-appropriate deployment in agricultural systems remains largely technique-centric. The present framework integrates analytical performance, sample complexity, and operational constraints to support the selection of fit-for-purpose methods across breeding, diagnostic, and dairy authentication contexts. Classical and advanced approaches, including polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP), allele-specific polymerase chain reaction (AS-PCR) and amplification refractory mutation system PCR (ARMS-PCR), high-resolution melting (HRM) analysis, sequencing-based methods, single nucleotide polymorphism (SNP) arrays, and digital polymerase chain reaction (dPCR), are comparatively evaluated not only in terms of sensitivity and throughput but also with respect to scalability, reproducibility, and decision risk. This framework provides a practical decision-support tool for aligning genotyping strategies with application-specific risk profiles, thereby improving reliability, transparency, and regulatory compliance in modern dairy systems.

Heat stress negatively affects bovine fertility and embryonic development, while cryopreservation imposes additional cellular stress that may impair embryo survival. The present study investigated whether controlled heat treatment applied during the early morula stage could influence subsequent blastocyst development, molecular stress responses, and post-thaw survival of bovine embryos. In vitro-produced bovine embryos were exposed to controlled thermal treatment involving gradual elevation in temperature from 38.8 °C to 40.0 °C followed by subsequent cooling. Blastocyst formation, post-thaw survival after slow freezing, and the expression of stress-associated genes and microRNAs were evaluated. Heat treatment did not significantly affect blastocyst formation rates or post-thaw embryo survival compared to the control groups. However, significant alterations in the expression of stress-associated markers were observed. The expression of HSP90AA1 and HSPB11 was significantly reduced following heat treatment, while cryopreservation induced increased expression of these markers compared to embryos exposed only to heat treatment. Significant alterations were also detected in the expression of several stress-associated microRNAs. These findings suggest that controlled heat exposure during the morula stage may induce molecular changes associated with the embryonic response to heat stress without compromising developmental competence or cryosurvival. The results contribute to a better understanding of the molecular mechanisms underlying embryonic responses to thermal and cryogenic stress and may support future optimization of embryo production and cryopreservation strategies.

The CSN2 gene encoding β-casein has gained increasing attention in dairy cattle breeding due to the global adoption of A2-oriented selection strategies. However, robust large-scale evidence assessing potential unintended effects on functional traits, particularly fertility, under intensive commercial conditions remains limited. This study evaluated whether selection for the CSN2 A2 β-casein variant is associated with biologically relevant differences in fertility traits in Holstein cows. Reproductive and genomic data from 7826 lactation records of 2773 Holstein cows collected between 2022 and 2025 in a large commercial dairy herd were analyzed. Fertility indicators included days open, number of services per conception, calving interval, first-service conception rate, and pregnancy by 100 days in milk. Mixed-effects models accounting for repeated lactations and cow- and sire-level clustering were applied, and predefined equivalence margins were used to distinguish statistical non-significance from biological irrelevance. Across all evaluated fertility traits, differences among CSN2 genotypes (A1A1, A1A2, and A2A2) were consistently small, biologically negligible, and well within predefined equivalence margins. Differences in days open were within ±2 days, and effect sizes for count and binary traits were close to unity. Parity and calving year significantly influenced reproductive performance, whereas no CSN2 genotype × parity interactions were detected. These findings indicate that selection for the CSN2 A2 β-casein variant does not compromise reproductive performance under intensive commercial management conditions. From a breeding and industry perspective, the results support the implementation of A2-oriented selection strategies without biologically meaningful adverse effects on fertility.

Allele-specific expression (ASE) reflects the unequal expression of the parental alleles and can imply functional variants in cis-regulatory elements. The conventional ASE detection methods often depend on the presence of heterozygous variants in transcripts or sequencing a large number of individuals, both of which are often limited. In this study, we present a family-based strategy for detecting ASE and potential cis-regulatory elements utilizing both RNA-seq and whole-genome sequencing (WGS) from a pedigree. Using a rabbit family consisting of two divergent parents and their eight offspring, we identified 913 ASE genes by analyzing inheritance patterns of gene expression levels. Expression was classified into three levels—high, medium, and low—and used to define seven distinct expression groups across the family (e.g., H_L: high in the mother, low in the father, and intermediate in the offspring). Many ASE genes lacked heterozygous exonic variants, and inference was achieved via RNA read count patterns. We also pinpointed conserved transcription factor binding sites (TFBS) with sequence variants showing similar inherited genotypic patterns (e.g., AAxBB), suggesting their regulatory roles as eQTLs. Differential gene expression (DEG) analysis between the parents highlighted some candidate genes related to meat production and quality traits. Our findings show that the family-based method using RNA-seq and WGS data is promising for exploring ASE and mapping possible eQTLs.

This study investigated the presence and levels of five key mycotoxins—aflatoxins (AFs), deoxynivalenol (DON), fumonisin B1 (FB1), ochratoxin-A (OTA), and zearalenone (ZEN) and its metabolite alpha-zearalenol (α-ZOL)—in liver samples from 19 golden jackals (Canis aureus) in southern Hungary. Golden jackals, as apex predators with a diverse diet encompassing both plant and animal matter, can serve as valuable bioindicators of environmental mycotoxin contamination. Genetic analysis confirmed the canid samples as coming from golden jackals, excluding the possibility of domestic dogs or hybrid individuals. All samples tested positive for at least three mycotoxins, with multiple mycotoxins frequently co-occurring. DON was detected in 95% of the samples, followed by FB1 (79%) and ZEN (42%). ZOL, AFs, and OTA were present in all samples. Significant differences in mycotoxin concentrations were observed between age groups and sexes for some mycotoxins. Specifically, adult males exhibited higher ZEN concentrations, adult females had higher OTA levels, and females generally showed significantly higher DON concentrations than males. For all investigated individuals, we found significantly higher concentrations of ZEN, alpha-ZOL, and OTA in adult samples. Malondialdehyde (MDA), an indicator of oxidative stress, was also measured and correlated with mycotoxin levels. Pareto analysis suggested a correlation between MDA and OTA/ZEN. These findings highlight the exposure of golden jackals to a range of mycotoxins in their natural environments, potentially through both plant and animal food sources, and underscore the potential of these canids as sentinels for mycotoxin contamination in ecosystems.

The European rabbit exists in the wild and has several highly bred domesticated forms. There are well-separated wild European rabbit populations, and intensive breeding has resulted in various forms and utilizations. In this work, we aimed to carry out an extended WGS-based population genomics study on several wild European rabbit populations and selected breeds. Utilizing multiple methods, we showed that although domestic and wild populations were clearly separated, there was evidence of admixture between them in France and Hungary. The populations showed various levels of inbreeding, with one of the Hungarian subpopulations having excess runs of homozygosity. We identified numerous variants fixed in either domestic or wild animals, two of which were found to be fixed at different alleles in the two populations. Some putatively selected regions did not overlap with any known genes in the rabbit genome, suggesting some importance to these intergenic sites. The enrichment of selected regions in certain types of transcription factor binding sites suggests a possible role for these regulatory elements during domestication. In addition, the new high-coverage rabbit whole-genome sequences may provide helpful material for further population genetics analyses.

Az in vitro embrió-előállítás (IVEP) 40 éves töretlen fejlődésének köszönhetően napjainkra egyre fontosabb szerepet tölt be a világ szarvasmarha-tenyésztésében. A petesejtek kinyerése, laboratóriumi körülmények között történő termékenyítése, az embriók tenyésztése, a krioprezerváció, illetve a laboratóriumi környezet terén elért fejlődés több millió genetikailag magas értékű egyed előállítását tette lehetővé. Gyakorlati alkalmazási lehetőségei mellett az in vitro embrió-előállítás (IVEP) számos kutatás alapját biztosítja. Segítségével a korai vemhességi időszak modellezhető, az embriófejlődés környezeti tényezői tanulmányozhatók, így olyan kérdésekre kaphatunk választ, melyek a tudományos és gyakorlati szakemberek számára is fontosak. Egy megbízhatóan működő embrió előállító rendszer kialakítása során sok technológiai lépés optimalizálására van szükség. Az elmúlt év során a szerzőknek sikerült kialakítaniuk egy olyan, jól működő rendszert, melyben sikeresen folyik a szarvasmarha embriók előállítása, így oktatási, kutatási feladatok megvalósítására alkalmas. A közleményben a szerzők bemutatják a rendszer kialakítását egészen a kezdetektől, ennek sikereit és buktatóit egyaránt, valamint beszámolnak az itt végzett 31 IVEP program eredményességéről. A kialakított rendszerben átlagosan a termékenyített petesejtek 69%-a osztódik, és 23,4%-a fejlődik beültethető állapotú hólyagcsíráig, így alkalmas ovum pick-up (OPU) IVEP programok megvalósítására is.

A genomic database of all Earth's eukaryotic species could contribute to many scientific discoveries; however, only a tiny fraction of species have genomic information available. In 2018, scientists across the world united under the Earth BioGenome Project (EBP), aiming to produce a database of high-quality reference genomes containing all ~1.5 million recognized eukaryotic species. As the European node of the EBP, the European Reference Genome Atlas (ERGA) sought to implement a new decentralised, equitable and inclusive model for producing reference genomes. For this, ERGA launched a Pilot Project establishing the first distributed reference genome production infrastructure and testing it on 98 eukaryotic species from 33 European countries. Here we outline the infrastructure and explore its effectiveness for scaling high-quality reference genome production, whilst considering equity and inclusion. The outcomes and lessons learned provide a solid foundation for ERGA while offering key learnings to other transnational, national genomic resource projects and the EBP.

The golden jackal ( Canis aureus ) is a reoccurring species in the centre of the Carpathian basin, in Hungary. In total, 31 golden jackal tissue samples were collected, from 8 white-coated, 2 black-coated and one mottled animal across Hungary. Sequences and fragment length polymorphisms were studied for white colour (MC1R), and for black coat colouration (CBD103). In each white animal, the most widespread mutation causing white fur colour in dogs in homozygous form was detected. Three animals were found to carry the mutation in heterozygous form. The two black golden jackals were heterozygous for the 3 bp deletion in CBD103 that mutation for black coat colouration in dogs, and one of them also carried the mutation causing white fur. None of the white animals showed signs of hybridization, but both the black and the mottled coloured individuals were found to be hybrids based on genetic testing. Kinship was found three times, twice between white animals, and once between a white animal and an agouti animal carrying the mutation of white coat. Our results confirm the findings that golden jackal – dog hybrids may occur without human intervention, and the detected mutation causing white fur colour in golden jackals could possibly be due to an early hybridization event.

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 ( H o ) and unbiased expected (u H E ) 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.

After the last glacial, the Carpathian Basin was repopulated from either eastward or northward colonisation routes for various species; one of these was the emblematic member of the European megafauna, the red deer, Cervus elaphus . We analysed 303 red deer individuals from the middle of the region, in seven Hungarian game reserves, at ten microsatellite loci (C01, C229, T26, T108, T123, T156, T172, T193, T501, T507), to investigate the genetic diversity of these subpopulations. We discovered high levels of genetic diversity of red deer subpopulations; allelic richness values ranging 4.99–7.01, observed heterozygosity 0.729–0.800, polymorphic information content 0.722–0.806, and Shannon’s information index 1.668–2.064. Multi-locus analyses indicated population admixtures of various degrees that corresponded to geographical location, and complex genetic structures were shown by clustering. Populations in the south-western and the north-eastern parts of the region formed two highly separated groups, and the red deer from populations in between them were highly admixed (in western Pannonia/Transdanubia, where the Danube flows into the Carpathian Basin). This pattern corresponds to the distribution of mitochondrial as well as Y-chromosome lineages. Assignment tests showed that a large fraction of individuals (29.4%) are found outside of their population of origin, indicating that the dispersal of red deer is rather common, which could be expected considering the life course of the species.

The golden jackal (Canis aureus) is a widespread opportunistic mid-sized canid, distributed throughout southern Asia, the Middle East and South-eastern and Central Europe. European populations have undergone significant population changes in recent decades. During the first two decades of the 21st century the size of the jackal populations increased in their distribution and abundance across Europe. Hungary and Romania apply similar game management practices, and the golden jackal appeared approximately at the same time in both countries. With the aim to determine the genetic structure and the origin of the expanding jackal populations, we analysed samples obtained from Romania and Hungary. Genotyped at 22 canine autosomal microsatellite loci (STR), samples were analysed using multivariate, Bayesian and landscape genetic methods. In the Romanian samples all loci were polymorphic with 3-12 alleles. The overall observed (Ho) and expected (He) heterozygosities were 0.552 and 0.647, respectively. In the Hungarian samples 20 out of 22 loci were polymorphic with 2-11 alleles. The overall observed (Ho) heterozygosity (0.561) was higher and the expected (He) heterozygosity (0.564) was lower than the corresponding Romanian value. Based on our clustering results, Romanian and Hungarian samples were separated into two different genetic clusters. These results show that smaller groups of golden jackals could become established in different regions following several episodes of colonization, possibly at different times and arriving from various locations.

In the food supply chain, quality control has a very important role in maintaining customer confidence. In the EU, food safety aspects are strictly regulated; however, composition requirements and standard control methods are generally undefined. The rapidly increasing wild boar population has a growing market share in venison or game meat production. Several methods have been described for species identification and control of composition in food products, but only some of these are suitable for routine measurements. The aim of our research was to design a rapid, reliable and simple PCR insertion/deletion (InDel)-based genetic tool suitable for species identification in food quality control laboratories. In total, 59 different swine (Sus scrofa) whole genomes were tested with bioinformatic tools to identify wild boar-specific insertions or deletions. Three independent InDels were suitable for marker development, multiplex PCR amplification and separation in agarose gel. Altogether, 209 samples of wild boar and ten other domestic pig breeds were taken for DNA extraction and validation of the three multiplexed InDel markers. Statistical analysis showed a very high combined predictive value (0.996), indicating the capability of the newly developed markers to detect wild boars with a probability over 99%. Breed assignment tests confirm that the InDel markers developed are suitable for rapid, sensitive and reliable identification of the wild boar meat content of food products. The use of the reported method in food quality control can mean a simple and cost-effective way to maintain consumer confidence and to support the competitiveness of fair producers.

In the Carpathian Basin the wild boar (Sus scrofa) belongs among the most important game species both ecologically and economically, therefore knowing more about the basics of the genetics of the species is a key factor for accurate and sustainable management of its population. The aim of this study was to estimate the genetic diversity and to elucidate the genetic structure and location of wild boar populations in the Carpathian Basin. A total of 486 samples were collected and genotyped using 13 STR markers. The number of alleles varied between 4 and 14, at 9 of the 13 loci the observed heterozygosity was significantly different (p < 0.05) from the expected value, showing remarkable introgression in the population. The population was separated into two groups, with an F-st value of 0.03, suggesting the presence of two subpopulations. The first group included 147 individuals from the north-eastern part of Hungary, whereas the second group included 339 samples collected west and south of the first group. The two subpopulations' genetic indices are roughly similar. The lack of physical barriers between the two groups indicates that the genetic difference is most likely caused by the high reproduction rate and large home range of the wild boars, or by some genetic traces' having been preserved from both the last ice age and the period before the Hungarian water regulation.

Microsatellites are widely applied in population and forensic genetics, wildlife studies and parentage testing in animal breeding, among others, and recently, high-throughput sequencing technologies have greatly facilitated the identification of microsatellite markers. In this study the genomic data of Cervus elaphus (CerEla1.0) was exploited, in order to identify microsatellite loci along the red deer genome and for designing the cognate primers. The bioinformatics pipeline identified 982,433 microsatellite motifs genome-wide, assorted along the chromosomes, from which 45,711 loci mapped to the X- and 1096 to the Y-chromosome. Primers were successfully designed for 170,873 loci, and validated with an independently developed autosomal tetranucleotide STR set. Ten X- and five Y-chromosome-linked microsatellites were selected and tested by two multiplex PCR setups on genomic DNA samples of 123 red deer stags. The average number of alleles per locus was 3.3, and the average gene diversity value of the markers was 0.270. The overall observed and expected heterozygosities were 0.755 and 0.832, respectively. Polymorphic Information Content (PIC) ranged between 0.469 and 0.909 per locus with a mean value of 0.813. Using the X- and Y-chromosome linked markers 19 different Y-chromosome and 72 X-chromosome lines were identified. Both the X- and the Y-haplotypes split to two distinct clades each. The Y-chromosome clades correlated strongly with the geographic origin of the haplotypes of the samples. Segregation and admixture of subpopulations were demonstrated by the use of the combination of nine autosomal and 16 sex chromosomal STRs concerning southwestern and northeastern Hungary. In conclusion, the approach demonstrated here is a very efficient method for developing microsatellite markers for species with available genomic sequence data, as well as for their use in individual identifications and in population genetics studies.