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Home » Archive » 2023

TDK conference 2023

Genetic sex detection of cervids by isothermal amplification
Bujtor Zsófia Krisztina - year 5
University of Veterinary Medicine Budapest, Department Animal Breeding, Nutrition and Laboratory Animal Science
Supervisor: Dr. Zenke Petra Zsuzsanna

Abstract:

Knowledge of the sexes and sex ratios of both domestic and wild animal species plays a central role in (wildlife) management and conservation biology, making it an intensively researched area. In Hungary, maintaining the optimal sex ratio of the populations of deer species, such as fallow deer, red deer, and roe deer, is also of great significance during the hunting season. Therefore, only a specific number, species, and sex of animals are allowed to be hunted during a given hunting season. However, there may be cases where the prescribed conditions are not met, and these are considered illegal hunting. In the cold stores of game processing facilities, the sex of eviscerated carcasses can no longer be identified based on external characteristics, but genetic testing can determine the sex of the animal, potentially revealing illegal kills, which are considered poaching.

The aim of our research was to develop a genetic method that allows for simple, fast, and on-site DNA-based sex determination of tissue samples from the three native deer species. As the basis for our investigations, we chose the loop-mediated isothermal amplification (LAMP) method, which allows for the rapid multiplication and detection of relevant DNA markers with minimal laboratory equipment and resources, in a cost-effective manner.

During our research, we first tested LAMP primers designed for conserved segments of the Amelogenin X and SRY genes, which can be universally used for the three deer species, on control DNA samples from known species and sexes. The results of the reactions were detected by the subsequent addition of an intercalating dye, and the composition and time of the reaction mixture were optimized. To simplify on-site sex determination, we developed a direct amplification method for meat samples prepared in NaOH solution. As expected, the Amelogenin X gene segment was detectable in both female and male samples, while the reaction targeting the male-specific SRY gene segment gave positive results for male-origin samples.

After validation steps, our developed method can be suitable for the rapid sex detection of fallow deer, red deer, and roe deer samples, contributing to the detection of potential illegal hunting activities and thus preserving the integrity of hunting.



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