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TDK conference 2020

DNA-based sex determination of Hungarian hunting game with antler for forensics
Bartal Balázs - year 3
University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science
Supervisor: Dr. Petra Zenke


There is a considerable amount of misusage related to hunting game with antler (red deer, fallow deer and roe deer). Cases, such as when the carcass is not from the timeframe it should be or does not match with the individual showed on the documentation. Verifying the sex of the eviscerated, morphologically unidentifiable carcasses using traditional methods can only happen in an uncertain manner and provide questionable results. Conservation biology has started using genetic markers in various species including trophy game for sexing. However, these studies providing a good foundation, do not meet the criterion set by forensic science, and cannot be universally applied to the Hungarian wild animal species with antlers. Exactly for this reason we would like to create an efficient, fast and widely useable sexing method for roe deer, red deer and fallow deer.

In our research, with the help of hunters, muscle and hide samples were gathered from these above mentioned games with known sex. We designed primers for the conserved parts of the Amelogenin X, the SRY and the Amelogenin Y markers, which produce less than 200 basepair PCR fragments. The laboratory-testing of these primers were done in a monoplex reaction on roe deer, red deer and fallow deer samples, then the nucleotide sequences of the specific products were confirmed by sequencing. After that, with a so called “touch-down” technique, a duplex PCR program was optimized, to be able to examine the X and Y markers simultaneously. The fragments amplified using fluorescently labelled primers were detected by agarose gel electrophoresis and then by a high precision capillary electrophoresis.

After the duplex PCR optimization process, as expected, both Y-chromosomal (SRY and the Amelogenin Y) and X-chromosomal (Amelogenin X) segments were detected in male samples, while only the Amelogenin X gene segment was detected in female samples. The developed genetic test was successfully used on a case related biological sample (red deer carcass with a questionable sex from the refrigeratory), which also shows the putative applicability in the field of forensic genetics.

To summarize, we were able to create a genetic method to determine the sex of red deer, fallow deer and roe deer with the same primer set, however, the validation process to reach the forensics standards is still in progress. Hopefully, our developed method could contribute credibly to the resolution of potential poaching disputes and reduce the number of illegal hunting activities.

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