Veterinary session

Microsatellite-based individual identification is becoming increasingly prevalent across various wildl animal species, including within the family of cervids (Cervidae), partly due to its applicability in forensic investigations. Examples include various forms of poaching, such as illegal trade in game meat and trophies, and road accidents involving wildlife fatalities. A set of 12 tetrameric microsatellite markers for European roe deer (Capreolus capreolus) has been previously developed and successfully applied to Swiss populations for individual identification. The primary aim of our research was to test this set of tetrameric microsatellite markers in Hungarian roe deer populations, to assess the genetic diversity of these populations, and to start building a Hungarian population database based on the genetic profiles.

We collected samples of skin or muscle tissue from professionally hunted animals (n=45) from three regions of the country. After DNA extraction and purification, the microsatellite regions were amplified by polymerase chain reaction. In order to achieve a cost-effective solution, we used fluorescently labelled universal primers to mark the products, in contrast to the original method. However, due to the technological limitations of the method, the markers were amplified in mono- or diplex form instead of the original multiplex reaction. After amplification, the length of the amplified DNA sequences was determined by capillary electrophoresis.

Although one of the loci (Capcap3.1) did not give valuable results due to significant by-product peaks, the set of 11 microsatellites was also adequate for the identification of Hungarian roe deer at the individual level. The markers tested were found to be polymorphic based on their allele numbers (NA=5-8), with observed heterozygosity (Ho) ranging from 0.40-0.84 and expected heterozygosity (He) ranging from 0.50-0.82 for all individuals tested. No significant differences were found between the genetic diversity of the three populations studied. The PI (probability of identity) value for all loci was 5.9×10ˆ(-11), which is in line with the requirements of the tests used in forensic genetic studies.

Based on our results, the marker set is suitable for the identification of roe deer in Hungary at the individual level based on the data of 11 markers that can be evaluated, but the use of the original set and its multiplex PCR protocol is recommended instead of using fluorescently labeled universal primers.