The fallow deer (Dama dama) represents significant game management value globally, and human activities are significantly impacting the species. Besides the positive effects, these activities can threaten its existence, health, and value. The aim of the authors was to develop a tetranucleotide microsatellite panel that could be clearly interpreted and used for genetic testing of fallow deer. Such a panel did not exist until now and could be particularly useful in the field of conservation genetics and forensics. A total of 99 tetrameric microsatellites, originally designed for related deer species, were tested on 20 fallow deer individuals from five Hungarian sampling areas. Original and newly designed primers were used to amplify the microsatellite regions using previously published or optimized PCR protocols. The lengths and sequences of specific amplicons were detected using capillary electrophoresis, and the rate of polymorphism was determined. Altogether, 80 markers provided PCR products of adequate quality and quantity. Among them, 15 markers proved to be polymorphic (2–5 alleles/locus), and 14 tetrameric markers were selected for further analysis. Statistical calculations showed that the selected polymorphic microsatellites can potentially enable key individualization in many areas of wildlife and population genetics, thus protecting the species.

The evidential value of an mtDNA match between biological remains and their potential donor is determined by the random match probability of the haplotype. This probability is based on the haplotype’s population frequency estimate. Consequently, implementing a population study representative of the population relevant to a forensic case is vital to correctly evaluating the evidence. The emerging number of poaching cases and the limited availability of such data emphasizes the need for an improved fallow deer mtDNA population databank for forensic purposes, including targeting the entire mitochondrial control region. By sequencing a 945-base-pair-long segment of the mitochondrial control region in 138 animals from five populations in Hungary, we found four different haplotypes, including one which had not yet been described. Our results, supplemented with data already available from previous research, do not support the possibility of determining the population of origin, although some patterns of geographical separation can be distinguished. Estimates of molecular diversity indicate similarly low mtDNA diversity (Hd = 0.565 and π = 0.002) compared to data from other countries. The calculated random match probability of 0.547 shows a high probability of coincidence and, therefore, a limited capacity for exclusion. Our results indicate that despite the overall low genetic diversity of mtDNA within the Hungarian fallow deer samples, a pattern of differentiation among the regions is present, which can have relevance from a forensic point of view.

The evidential value of an mtDNA match between biological remains and its potential donor is determined by the random match probability of the haplotype. This probability is based on the haplotype’s population frequency estimate. Consequently, implementing a population study representative of the population relevant to the forensic case is vital to correctly evaluating the evidence. The emerging number of poaching cases and the limited availability of such data emphasizes the need for improved fallow deer mtDNA population databank for forensic purposes, including targeting the entire mitochondrial control region. By sequencing the whole mitochondrial control region in 138 animals from five populations in Hungary we found four different haplotypes, including one which has not yet been described. Our results, supplemented with data already available from previous research do not support the possibility of determining the population of origin, although some patterns of geographical separation can be distinguished. Estimates of molecular diversity indicate approximately similarly low mtDNA diversity (Hd=0.565 and π = 0.002), compared with data from other countries. The calculated random match probability of 0.547 shows a high probability of coincidence and, therefore, a limited capacity for exclusion. Consequently, it may be necessary to search for polymorphic sites on other mitochondrial gene sections or to sequence the entire mitogenome of fallow deer.

Determining sex in Cervidae species is crucial for wildlife management, conservation, breeding programs, disease surveillance, and forensic investigations. While existing genetic sexing methods have proven effective, they often involve time-consuming and resource-intensive processes. In response to the growing need for a more efficient and cost-effective technique, this study aimed to develop a novel loop-mediated isothermal amplification (LAMP) method for sex detection in various deer species. We designed LAMP primers specific to Cervidae based on sex chromosome-located genes, AmelogeninX, and SRY. Analytical specificity was confirmed using 108 samples from red deer, roe deer, and fallow deer, and species specificity was also tested in other deer and mammal species. Sex determination results were fast (40 min) and accurate (100% reliability). This technique has the advantage of allowing researchers to identify the sex of different deer species using the same method. Using the isothermal amplification properties of LAMP, this new method provides a simple, quick, and accurate tool to help researchers, wildlife managers, and forensic investigators with important sex-related questions in cervid populations.

The fallow deer (Dama dama) of Hungary has excellent value to our country due to its game meat and antler trophies. As an attempt to aid law enforcement against illegal activities, such as poaching, illegal trading, and in other cases like traffic accidents, we aimed to develop a genetic marker set suitable for individual identification. During our research, 28 microsatellite markers on 15 fallow deer samples from two different populations were tested. Four microsatellites were found to be polymorphic, each with two or three alleles. Based on our current results, Hungarian fallow deer populations show low genetic diversity. This is in agreement with previous studies conducted on the species and is probably a direct result of the species’ past extinction from the most of Europe during the Pleistocene and later its human-mediated reintroduction to most of its current range. The low number of polymorphic markers presents the need to include additional markers.fallow deer

A simple, fast, and cost-effective organic solvent-based protocol was developed for DNA extraction from deer antlers and prepared trophy skulls, eliminating the need for commercial kits or cryogenic grinding. The method combines bead-based mechanical homogenization with a 4 h enzymatic digestion in EDTA buffer containing N-lauryl sarcosine and Proteinase K, followed by phenol–chloroform–isoamyl alcohol purification and centrifugal filtration. DNA quality and quantity were evaluated using agarose gel electrophoresis, Qubit fluorometry, and Nanodrop spectrophotometry. The protocol was tested on 60 samples, comprising 30 antlers and 30 pedicle parts from prepared trophy skulls of roe deer (Capreolus capreolus), fallow deer (Dama dama), and red deer (Cervus elaphus). To assess suitability for downstream applications, species-specific microsatellite markers were amplified using multiplex PCR, successfully generating complete genotypes from all 60 samples. These results, along with a demonstrated case study, confirm that the developed protocol provides high-quality DNA suitable for molecular genetic investigations, enabling reliable genotyping from small amounts of both antler and processed trophy materials in forensic and conservation contexts.