With growing demand for pedigree verification and breed management in horses (Equus caballus), reliable paternal lineage tools are essential. Y-chromosomal STRs (Y-STRs) have advantages over autosomal STRs due to paternal inheritance and lack of recombination. However, few validated loci and no standardised efficient genotyping systems limit their use. Current methods often require multiple reactions, increasing cost and labour. Thus, identifying informative Y-STR loci and developing a multiplex PCR system for cost-effective paternal lineage analysis is urgently needed. To identify informative horse Y-STR loci and develop a multiplex PCR system for paternal lineage tracing, breed conservation, pedigree verification, and forensic settings. Experimental study. Y-STR loci were screened from the horse reference genome based on repeat motif, number, and location. Primers were designed and tested on stallions and mares to confirm Y-specificity. Loci with stable, male-specific amplification were combined with known markers into a multiplex PCR system. Sensitivity, repeatability, reproducibility, and performance were evaluated, and locus polymorphism was assessed in Debao ponies, Mongolian horses, Thoroughbreds, and Arabian horses. Twenty candidate loci were screened, and 15 of them showed clear, reproducible male-specific amplification. Three novel polymorphic loci were identified and combined with 5 known markers into 2-plex and 6-plex panels. Most loci were polymorphic in Debao ponies (PIC: 0.000-0.575) and Mongolian horses (PIC: 0.000-0.375), while less polymorphism was detected in Thoroughbreds (PIC: 0.000-0.099) and Arabian horses (PIC: 0.000-0.099). The optimised multiplex PCR system demonstrated high sensitivity, repeatability, and compatibility. Reproducibility and stability across labs and platforms require further validation. Fifteen new Y-STR loci were identified, and 3 of them were polymorphic. Novel optimised multiplex PCR systems were developed for the identification of horse paternal lines, providing reliable tools for equine genetic study, breeding, and forensic applications.

Cattle are frequently involved in civil disputes and offenses. Currently, the paternity testing of cattle is mainly done by analysis of dinucleotide STR (di-STR) or SNPs. However, the di-STR marker usually produces many stutter artifacts that affect the accurate genotyping, and the polymorphic level of SNPs is not high enough. To overcome these limitations, we developed and validated a novel integrated next-generation sequencing (NGS) panel for cattle. It comprises 33 reported di-STRs and 30 newly identified tetranucleotide STRs (tetra-STRs), the mitochondrial displacement loop (D-loop) region, as well as cytochrome b (CYTB) and recombination-activating gene 1 (RAG1). The results showed a high genotyping success rate (99.6% on average) and balanced amplification efficiency across most STR loci. In addition, the panel's average genotyping consistency rate reached 96.4% between blood and hair samples. The total discrimination power and cumulative probability of exclusion for duos of this panel were 1-1.8×10-13 and 1-1.7×10-8, respectively. Application in paternity testing caseworks demonstrates its applicability and reliability for resolving paternity disputes. Overall, this study provided a cattle profiling NGS panel with a substantial advancement over current capillary electrophoresis (CE)-based systems, offering superior accuracy, multiplex capability, and efficiency for forensic genetic testing in cattle.

Amelogenin is the most common marker in Autosomal STR kits to detect the gender of the sample in forensic DNA case analysis. An essential component of autosomal STR kits for DNA fingerprinting in forensic case investigation is Amelogenin-based gender analysis. It becomes challenging to report the gender of a person when a DNA sample collected from a phenotypically male individual shows a dropout of the Amelogenin Y-allele and Y-STR markers can be incorrectly interpreted as those of a female DNA sample. Therefore, a detailed analysis is crucial for confirming the dropout of the amelogenin Y-allele in such cases, as it is of utmost importance in forensic DNA case reporting. DNA isolation and quantification were done using standard protocols. ABS 3500 Genetic Analyzer (Applied Biosystems, USA) is used for genotyping. Autosomal STR amplification was performed using the VeriFiler Plus Amplification kit (Applied Biosystems, USA) and PowerPlex Fusion 6C system kit (Promega, USA). For Y STR amplification, Investigator Argus Y-28 QS Kit (Qiagen, Germany) and Y Filer Plus PCR Amplification kit (Applied Biosystems, USA) were utilized. Out of 317 forensic DNA cases examined from the period from 2021 to 2023, dropout of Amelogenin Y allele and Y STR markers in autosomal STR kits was detected in N=02 forensic DNA cases. The DNA samples showed dropout in the p arm STR and null allele at the AMEL-Y locus when analyzed using autosomal STR kits. Further examination with Y-STR kits showed no allelic data on six loci in the Yp11.2 region in addition to the Amelogenin Y locus. A careful consideration should be given to the null allele based on the population data to avoid wrong gender reporting using only autosomal STR kits having amelogenin and proximal Y STR markers having loci on the p arm of Y chromosome.

DNA forensic investigations can sometimes produce unexpected results, even in seemingly straightforward cases. One such case involved a paternity test where an alleged father sought confirmation of his biological relationship with a young girl. After two prior examinations produced contradictory results, the test was repeated at the DNA Laboratory of the Institute of Scientific Police in Tirana. Samples included three buccal swabs from the alleged father, one from the mother, and one from the child. Sixteen autosomal STR loci and AMELO were analyzed using PCR, followed by capillary electrophoresis on a Genetic Analyzer 3500 HID. The alleged father’s samples revealed a mixed DNA profile: three loci with three alleles, three loci with four alleles, and ten loci with two alleles. His medical history disclosed a prior bone marrow transplant, which explained the chimeric DNA pattern. This represents the first documented case of chimerism affecting DNA-based paternity testing in Albania. The case highlights how chimerism can lead to false exclusions in parentage testing and complicate genetic investigations. Awareness of medical history and expert consultation are essential to reduce misinterpretation and ensure reliable conclusions in such cases.

The present study, “Judicial Awareness and Technical Literacy in Evaluating DNA Reports: A Study among Bhopal District Court Judges and Prosecutors,” examines the extent to which judicial officers, prosecutors, and defense lawyers comprehend and apply DNA-based forensic evidence in criminal trials. The research seeks to evaluate the level of scientific literacy, interpretational competence, and confidence in handling DNA profiling reports as admissible evidence under Section 45 of the Indian Evidence Act and relevant provisions of the Bharatiya Nagarik Suraksha Sanhita (BNSS) 2023. A structured questionnaire was administered to 40 respondents, comprising 10 judges, 15 public prosecutors, and 15 defense lawyers of the District Court, Bhopal. The data were analyzed using descriptive statistics and ANOVA to identify variations in awareness and reliance levels across professional groups and years of experience. Findings indicate that 80% of judges, 73% of prosecutors, and 60% of defense lawyers considered DNA profiling a “highly reliable” form of scientific evidence. However, only 55% of respondents demonstrated adequate understanding of critical technical concepts such as allelic frequency, STR loci, and chainof-custody procedures. ANOVA results revealed significant differences (p < 0.05) in interpretational confidence between judges and defense lawyers, highlighting disparities in technical familiarity. Moreover, 68% of participants supported the inclusion of forensic science modules in judicial and legal education programs. The study concludes that while acceptance of DNA evidence is generally high across Bhopal’s judicial community, technical literacy remains uneven. It recommends structured capacitybuilding initiatives, joint forensic–legal training workshops, and standardized interpretation guidelines to enhance evidentiary precision and promote scientifically informed judicial decision-making.

The probabilistic analysis performed in DNA kinship cases is affected by factors such as the number of loci analyzed, the STR allelic relative frequency dataset, and the population genetic characteristics of the individuals involved in the analysis. The Lebanese population shows high rates of endogamy and consanguinity, which challenges the power of discrimination of DNA profiles and is a potential source of misleading conclusions. The present study aims to evaluate the probability of false positives in relationship testing, especially siblingship, avuncular, and grandparent-grandchildren relationships, when cases are subject to different constraints, including DNA profile sizes (15 vs. 23 STR loci) and inbreeding coefficients (F ST). A total of 126 794 kinship comparisons were performed for each type of kinship test using a Lebanese database of 505 unrelated healthy individuals. To do so, DNA analysis results were assessed under a pair of propositions involving the stated relationship link between the two individuals vs. unrelatedness, offering quantification through the likelihood ratio (LR), a widely accepted metric to assess the value of scientific results. Among the simulated cases, false associations for full siblingship relationships were observed in 1 995 (1.6%) instances using a DNA profile size of 15 STRs (LR > 1, or log(LR) > 0). False associations decreased to 340 (0.3%) cases when the DNA profile size was increased to 23 STRs, and to 299 (0.2%) and 91 (0.07%) cases using the co-ancestry coefficient for the Lebanese population (0.002) and that recommended by the National Research Committee (0.02), respectively. Simulations of half siblingships (representing half-siblingship, avuncular, and grandparent-grandchildren relationships) showed similar results. Conversely, 41 full biological siblingship cases, 24 true full-siblingship laboratory cases, and 124 true half-siblingship laboratory cases were used as controls to assess the effects of implementing the conditions applied in the simulations (profile size and co-ancestry coefficient (F ST) values) to lower false associations. Results for the positive controls showed that the implementation of the National Research Committee-recommended F ST value of 0.02 decreased the LR by an average of 102 for 15-STR DNA profiles, and by an average of 104 for 23-STR DNA profiles, which did not lead to misleading results. However, the use of a DNA profile size of 23 STRs with the Lebanese co-ancestry coefficient showed the most meaningful results. This study could help to set national recommendations for the interpretation of kinship testing in the Lebanese population.

Background. Modern molecular genetic technologies make it possible to monitor genetic resources, both at individual and population levels, and reduce the time of genetic improvement of herds. Purpose. The given research aims to study the polymorphism of microsatellite loci in Limousine beef cattle bred in the Republic of Bashkortostan. The objectives include analysing the polymorphism of microsatellite loci in Limousine cattle, produced by absorptive crossing of Limousine bulls with dual-purpose cows (Simmental and Bestuzhev) and investigating the genetic structure and indicators of genetic diversity of Limousine cattle subpopulations. Materials and methods. The research objects are young Limousine cattle of absorptive crossing grown in the limited liability farm "Miasnoi soyuz bashkirskikh proizvoditelei (Meat Union of Bashkir Producers) and SEC Yaroslavsky. The research was conducted in the genetic laboratories of the Bashkir State Pedagogical University named after M. Akmulla, the Federal State Budgetary Educational Institution of the Higher Education the Russian State Agricultural University named after K.A. Timiryazev and Bashkir Agricultural Research Institute of the Ufa Federal Research Centre of the Russian Academy of Sciences. Results. The analysis of 16 STR loci in Limousine cattle DNA revealed 116 alleles in group I, where the maternal foundation was of the Simmental breed, and 74 alleles in group II, where the maternal foundation was of the Bestuzhev breed. The average number of alleles per locus was 7.25 in group I, and 4.63 in group II. The mean number of effective alleles per locus in groups I and II was 4.14 and 3.27, respectively. Group I exhibited high Shannon index values in the loci TGLA227, BM2113, TGLA53, CSSM66, and INRA023. The observed and expected heterozygosity levels showed no significant differences. The spatial arrangement of genotypes in the principal coordinate system indicated that the studied subpopulations represent well-consolidated groups, with the preservation of individual genetic diversity in individuals, united by common ancestry and belonging to the same breed. Conclusion. The obtained results can be used for the rational use of genetic resources of cattle of the Limousine breed and the development of genetically based breeding programs.

Latent fingerprints are a valuable yet challenging source of both human and microbial DNA, especially when ridge patterns are of poor visual quality with little to no identification potential. The efficiency of DNA collection and extraction depends on the choice of the sampling technique and the specimen conditions. This study compared two swab types for collecting and extracting human and microbial DNA from the fingers and fingerprints of two participants: 4N6FLOQSwabs® Genetics (Copan Italia) and standard cotton swabs (Dealmed Cotton-Tipped Sterile Wood Applicators). Results revealed higher DNA recovery for one participant regardless of swab type, indicating variability likely due to individual differences in natural cell/DNA shedding. Microbial DNA recovery ranged from 100% (from fingers) to 30% (from fingerprints), implying a loss of DNA during deposition, with 4N6FLOQSwabs® generally outperforming cotton swabs. Total DNA recovery from fingerprints was ~57% of that from fingers, comparable between both swab types, though 4N6FLOQSwabs® yields were higher overall. STR profiling via direct PCR with MicroFLOQ® showed that 4N6FLOQSwabs® performance was also superior, successfully identifying STR loci from fingers in 74% of cases, compared to 54% with cotton swabs. This figure was similar when extracting DNA from fingerprints (~40%). This comparative study underscores the importance of selecting the appropriate swab type for optimal DNA extraction from fingers and latent fingerprints in forensic investigations.Key Points 4N6FLOQSwabs® more effectively collects DNA from fingers and fingerprints than cotton swabs.Direct PCR with MicroFLOQ® on 4N6FLOQSwabs® provides higher DNA quality as shown by STR profiling.4N6FLOQSwabs® and cotton swabs detect no influence of handedness in DNA recovery.

In this study, 137 pairwise relationships representing four major relationship categories involving 49 Turkish individuals from four families were analyzed to evaluate the potential gain in the statistical power associated with likelihood ratios (LR) when using sequence-based versus length-based genotyping methods over the same STR loci coverage. To this end, the MPS Precision ID GlobalFiler NGS STR panel Kit and CE GlobalFiler™ PCR Amplification Kit were used. MPS-based analysis revealed the presence of 37 STR DNA sequence variations and / or the presence of 26 STR DNA sequence flanking region SNPs compared to the 150 unique alleles obtained with CE-based genotyping. Considering that most kinship LR calculation software do not readily take into consideration STR DNA sequence variants and STR DNA sequence flanking region SNP data that becomes available during MPS-based genotyping, an alphanumeric allele re-coding system was implemented to incorporate such additional STR isoallelic data to the already available allele calls. Over all the four major relationship categories analyzed, a significant increase in the mean combined LR (cLR) was observed when going from CE-based to MPS-based typing, whereby a 78.08 to 7,864,630.60-fold increase was noted. More specifically, in 134 out of the 137 pairwise relationships analyzed, MPS-based cLR values were higher than those calculated using CE-based data. While the mean cLR was >1,000 for three out of the four major relationship categories when using CE, the only exception being the third degree relationships, the mean cLR was >1,000 for all the four major relationship categories when using MPS. Notably, the mean cLR obtained for the third degree relationships was 47.61 with CE and 3,717.31 with MPS. In comparison with CE-based genotyping, when fully taken into account as proposed in the current study, the DNA sequence variation data afforded by MPS-based genotyping led to a statistically significant gain in terms of cLR values obtained. The use of MPS for cLR calculations had the most impact for both the second and third degree relationships, the two complex / distant type analyzed, hence further underscoring the prospects for MPS in kinship analysis. While the current study demonstrated that cLR is likely to increase substantially upon going from CE to MPS genotyping over the same loci coverage for a given case, when the additional DNA sequence variances are also taken into consideration, further increases are expected due to the more diverse type of forensic markers and even wider loci coverages used by MPS kits.

The aim of the study is to assess genetic variability in the population of Yakut horses of the Yana intrabreed type using an extended panel of STR loci, as well as to study the polymorphism of the ASIP, MC1R and STX17 genes that control the characteristics of hair pigmentation. The total number of animals examined was 55. Horses were tested using 25 autosomal STR loci (AHT4, AHT5, ASB2, ASB17, ASB23, HMS1, HMS2, HMS3, HMS6, HMS7, HTG4, HTG6, HTG7, HTG10, TKY279, TKY287, TKY321, TKY343, TKY344, TKY294, TKY325, TKY333, TKY341, TKY394, VHL20) and one locus located on the X chromosome (LEX3). Analysis of the genetic structure of the Yana horse population by STR loci showed a high level of genetic variability. The total number of alleles found in 26 studied loci was 208, of which 67 alleles were rare. The average number of effective alleles was 4.69 per locus. The average values of expected (He) and observed (Ho) heterozygosity, taking into account the 25 studied autosomal loci, were 0.762 and 0.761, respectively. A study of the polymorphism of genes controlling hair pigmentation showed that the frequency of the mutant variant of the ASIP gene in the group of Yana horses was 0.500, the frequency of the mutant variant of the MC1R gene was 0.327. The dominant mutation of the STX17 gene, associated with the gray phenotype, was found in 56.36 % of the tested animals. Taking into account the results of testing for 3 genes controlling pigmentation, 14 different genotype variants were identified in the studied population.

New population data for 21 STR loci in the Melanau and Murut of Borneo.

Human immunodeficiency virus (hereafter, "HIV") and hepatitis B virus (hereafter, "HBV") are prevalent infectious pathogens. In many countries, criminal law applies to those who intentionally transmit or expose others to these two dangerous pathogens. Viral infection is a significant characteristic in forensic investigations, and there are few reports on the simultaneous detection of DNA viruses, RNA viruses, and human short tandem repeats (hereafter, "STRs") in forensic laboratories. We developed a novel multiplex assay system that simultaneously identifies HIV-1, HBV, and STRs via capillary electrophoresis (hereafter, "CE"). This system demonstrated high species specificity, detecting HIV-1 RNA and HBV DNA at concentrations as low as 10 IU/mL. Full STR profiles were generated using 62.5 pg of template DNA. The combined power of discrimination of the 16 STR loci met the requirements of forensic applications. The comprehensive validation results confirmed that the multiplex assay system, which can provide qualitative results for three types of targets in a single tube via a one-step amplification process, is valuable for forensic applications. This system can offer additional individual characteristics (viral infections) for forensic DNA analysis, thus improving the efficiency of forensic investigations.

Superfecundation, the fertilization of two oocytes by different spermatozoa within the same ovulatory cycle, can result in monopaternal or heteropaternal dizygotic twins. While monopaternal superfecundation is more common, heteropaternal superfecundation is rare and typically seen in disputed paternity cases. This study presents a case of heteropaternal superfecundation confirmed through forensic DNA analysis and reviews its occurrence in existing literature. A forensic investigation was conducted in a court-ordered paternity case involving dizygotic twins, their mother, and an alleged father. Buccal swab samples were collected and analyzed using multiplex amplification of 19 STR markers and the amelogenin locus. A second DNA test confirmed the results. Additionally, a dataset of 2,679 paternity tests over 10 years was examined to estimate paternity exclusion rates in twin cases. Genetic analysis confirmed the alleged father's paternity of twin 1 but not twin 2, with 14 out of 19 STR loci showing absent alleles in twin 2. The 10-year dataset showed 553 paternity exclusions (20.64% of cases), with 31 involving twins, of which one case (3.23%) was identified as heteropaternal superfecundation. No significant difference was found between paternity exclusion rates in twin and non-twin cases. This case underscores the value of forensic genetic testing in detecting heteropaternal superfecundation, a rare occurrence with legal and social implications. Advances in DNA analysis may lead to more frequent identification of such cases.

The snails of the genus Helicopsis belong to steppe species, many of which are listed in the protected lists of European countries. In this work, based on the sequencing of the COX1 mitochondrial gene, the species belonging to eight populations of snails from the genus Helicopsis living in the south of the Central Russian Upland have been identified. These populations were assigned to H. lunulata, H. filimargo, and H. hungarica, and the average genetic distance between them was 0.11±0.01. Next, we analyzed the variability of seven microsatellite (STR) loci developed by us for H. lunulata in one population of H. filimargo, one population of H. hungarica and five populations of H. lunulata in the south of the Central Russian Upland. Both loci specific to H. lunulata and universal for all studied species have been identified. In H. lunulata, six STR loci turned out to be polymorphic and had from 2 to 11 alleles per locus. A total of 28 alleles were identified at all loci in H. lunulata, 14 of which were private and did not occur in more than one population. Molecular Dispersion Analysis (AMOVA) confirmed the high genetic differentiation of H. lunulata populations (Fst=0,425). At the same time, the genetic diversity of H. lunulata in the study area based on STR loci turned out to be higher than in the case of using allozyme loci. The structure of genetic variability calculated on the basis of STR loci turned out to be less pronounced compared to the use of allozyme loci as genetic markers (Fst=0,700). All this confirms the greater selective neutrality and greater diversity of microsatellite loci compared to allozymes. The data obtained can be used to assess the genetic diversity of H. lunulata. populations.

To investigate the genetic differences among different populations based on 13 autosomal STR loci in CODIS core. Data of 13 autosomal STR loci (CSF1PO, FGA, THO1, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11) were collected from 95 populations in scientific journals between 1999 and 2021, soursed from the PubMed database, which had been published. Allele frequencies of loci were sorted out and forensic genetic parameters including gene differentiation coefficient (Gst), total heterozygosity (Ht), subpopulation heterozygosity (Hs) values, and Nei's DA genetic distance were calculated. Principal component analysis, phylogenetic tree, and multidimensional scale analysis were conducted to assess population genetic structure. A total of 265 alleles were detected at the 13 STR loci in these 95 populations. The mean values of Gst, Ht, and Hs were 0.023 247, 0.797 915 and 0.779 365. Population genetic analyses reflected significant differences among populations from Asia, Africa and Europe. In Asian populations, there was a certain degree of distinction between mainland and island populations; the Han population showed a certain degree of distinction with surrounding populations in mainland; while within the Han population, there were two distinct clusters formed by the northern Han and the southern Han. The 13 autosomal STR loci in CODIS core demonstrate potential value for population identification across different groups, and may be used for the differentiation of ethnic groups, among different continental populations.

To evaluate the forensic application value of used dental floss as a source of biological evidence for individual identification by analyzing the effects of dental floss sample collection methods, DNA extraction methods, preservation conditions, and sampling sites on the success rate of STR typing. Dental floss samples were collected using three techniques: direct cutting, cotton swab wiping, and flocked swab wiping, respectively. DNA was extracted respectively by the Chelex, spin column-based and magnetic bead-based methods. DNA quantification and STR typing were performed using the Qubit kit and FGI HumDNA Typing kit (Platinum), respectively. Storage environments (temperature and humidity, ultraviolet radiation) and sampling locations (the floss part, the handle part) on DNA quantity and STR typing were evaluated. Through conducting a statistical analysis of three key indicators of average DNA mass concentration, STR locus detection rate, and typing accuracy rate, the direct cutting method demonstrated the highest efficacy, followed by cotton swab wiping mothed, and the flocked swab wiping method had the lowest efficacy. Direct cutting yielded an average DNA mass concentration greater than (4.94±1.87) ng/μL, with STR locus detection and accuracy rates of 100%. Bead-based DNA extraction method produced superior DNA concentration and quality compared to spin column-based and Chelex methods, regardless of whether the sampling technique used. Preservation conditions had a significant impact on the DNA analysis of samples. Particularly, the STR typing accuracy of samples preserved at 55 ℃/50%RH for 35 days dropped to (81.82±12.31)%, and that of samples exposed to ultraviolet radiation for 12 h dropped to (55.46±34.31)%. DNA concentration from the handle part of dental floss was extremely low, with an STR typing accuracy of only (30.91±27.35)%. Using cotton swabs to wipe or directly cutting the thread of dental floss samples, and combining this approach with the magnetic bead method for DNA extraction, can best guarantee the concentration and quality of DNA. In addition, samples should be stored in low-temperature, low-humidity environment, protected from light and ultraviolet radiation.

Population data of 23 Y chromosome STR loci for Kyrgyz population from Kyrgyzstan

The Eurasian lynx (Lynx lynx) is listed in CITES Appendix II and is protected under the Bern Convention and the EU Habitats Directive, yet it remains a frequent target of wildlife crime, highlighting the urgent need for reliable identification methods. This study focuses on determination and DNA quantification of the Lynx spp. using quantitative real-time PCR (qPCR). The Llynx Qplex quantification multiplex system effectively distinguishes Lynx spp. from other Feliformia species by targeting mitochondrial and nuclear markers. Additionally, we present the results of the developmental validation of the Llyn STRplex system for individual identification and databasing using six STR loci. This study followed ISFG recommendations for non-human DNA testing and developmental validation guidelines. Both systems demonstrate high sensitivity (5 pg genomic DNA for Llynx Qplex and 30 pg of mtDNA for Llyn STRplex) and high specificity to Lynx spp., confirmed by testing against 16 related Feliformia species. Robustness was evaluated, showing sensitivity to temperature variation, and both repeatability and reproducibility were successfully tested across replicates and conditions. Given that forensic casework often involves degraded and limited biological material, molecular tools must be both sensitive and specific to ensure accurate results. Developing precise and efficient tools is essential for supporting investigations of wildlife crime involving the Eurasian lynx, as well as efforts aimed at conserving the species.

Background The agropastoral communities of Coquimbo, Chile, are characterised by their goat herding-based livelihoods, admixed ancestry, and transhumant mobility. Aim To explore the impact of these features on genetic diversity and interactions with neighbouring populations. Subjects and methods Genotypic polymorphisms of 15 STRs were analysed in 466 individuals from 15 communities. Forensic parameters were estimated. Genetic structure was assessed using RST, Nei’s distances, MDS, dendrograms, and STRUCTURE, with 23 reference populations from Chile, South America and globally. Results A total of 158 alleles were observed, with frequencies ranging from 0.0011 to 0.5172. CSF1PO, D18S51, and Penta E showed deviation from Hardy-Weinberg equilibrium. The panel demonstrated high forensic performance (combined power of discrimination CPD > 0.999999999, combined power of exclusion CPE = 0.99999713817). No clear genetic structure was found within the Coquimbo communities. Regionally, Coquimbo clustered with northern Chile and north-west Argentina. Globally, it resembled other South American admixed populations, slightly differentiated from those from other regions. Conclusions The STRs analysed show high forensic potential, low genetic structure within the agropastoral communities, and important similarities with populations in northern Chile and north-west Argentina, supporting the relevance of trans-Andean mobility in shaping their genetic landscape.

For genetic characterization of dogs of Cavalier King Charles Spaniel breed, a comparative analysis of genotypes by STR loci of DNA of adult dogs was performed. The animals were divided into four groups distinguishable by color. From 1 to 5 allelic variants per locus were revealed with an average number of alleles of 2.9 per locus. Three alleles were found in the loci AHTk211 1, CXX279 1, REN169O18 1, INU005 1, AHTh171 1. Four alleles were found in the loci AHTh260 1, REN54P11 1, AHT137 1, AHTk253 1, REN162C04 1. The locus REN169D01 was characterized by the maximum number of alleles (5 alleles) with the number of effective alleles of 3.0. The fi xation index F (Fisher-Ryder criterion) in different groups of dogs of Cavalier King Charles Spaniel breed varied from –0.636 to –0.091. The negative deviation of the fixation index indicated an excess of heterozygotes identified in the four studied groups of dogs. Analysis of the genetic identity indices according to Ney showed that the level of genetic similarity between the groups of dogs ranged from 0.793 to 0.902. For example, the 2nd group was identical to the 3rd group (0.857), and the 1st and 4th groups were identical (0.793). Comparative analysis showed that the 1st and 2nd groups of dogs had a high degree of genetic identity between themselves (0.828), 2nd and 4th (0.829), 3rd and 4th (0.821), while the 1st and 3rd, 1st and 4th groups were more distant according to Nei. Thus, there is a different genetic identity between the studied groups of dogs by color, which must be taken into account in further breeding work with the breed Cavalier King Charles Spaniel.