Volume: 5, 2026
5th International PhD Students’ Conference at the University of Life Sciences in Lublin, Poland:
ENVIRONMENT – PLANT – ANIMAL – PRODUCT
Abstract number: A012
DOI: https://doi.org/10.24326/ICDSUPL5.A012
Published online: 22 April 2026
Effectiveness of Percoll density gradient separation of Arabian stallion semen in selecting spermatozoa with preserved DNA integrity
Kornelia Gala*1, Zuzanna Tarasek1, Artur Gurgul2, Agnieszka Kozlowska3 and Monika Bugno-Poniewierska1
1 Department of Animal Reproduction, Anatomy and Genomics, University of Agriculture in Krakow, Poland
2 Center for Experimental and Innovative Medicine, University of Agriculture in Krakow, Poland
3 Michałow Stud Farm, DW766, 28-411 Michałow, Poland
* Corresponding author: kornelia.gala@student.urk.edu.pl
Arabian horses are a highly valued breed worldwide. They are used across a wide spectrum of applications. Despite their high phenotypic value, some stallions produce semen with reduced quality parameters. Semen quality is characterized by considerable variability, both between individuals and among ejaculates from the same stallion. Consequently, this limits the applicability of assisted reproductive techniques, particularly when only a single semen dose with suboptimal parameters is available. In equine reproduction, one of the most commonly used methods to improve ejaculate quality is density gradient centrifugation (DGC). This technique enables the separation of a fraction enriched with spermatozoa of high physiological and morphometric potential from dead and damaged sperm cells, as well as from other cells and seminal plasma. The assessment of basic semen parameters is not always sufficient to predict high insemination success and proper embryo development. Sperm DNA integrity may also serve as a fertility marker.
Due to the variability in stallion semen quality and the limitations in its application in reproductive biotechnologies, the aim of the present study was to evaluate whether separation of ejaculate using a density gradient reduces the proportion of spermatozoa with DNA fragmentation. The experimental material consisted of semen collected from Arabian stallions (n=6). Semen samples were subjected to density gradient centrifugation using Percoll (40% and 80%). DNA fragmentation was assessed both before centrifugation and after separation in each fraction, with approximately 5000 spermatozoa analyzed per sample. The results demonstrated significant differences in the proportion of spermatozoa with DNA fragmentation between the analyzed fractions. The mean level of DNA fragmentation before centrifugation was 16.71 ±6.54 and significantly decreased following the use of the 80% Percoll fraction (2.47 ±1.33). Repeated measures analysis of variance confirmed a significant effect of the applied fraction on DNA fragmentation levels. Conover’s post hoc test with Holm correction revealed significant differences between pre-centrifugation samples and both the 80% (p<0.001) and 40% (p=0.001) fractions, as well as between the two Percoll fractions (p=0.018). The obtained results indicate high effectiveness of Percoll gradient separation in reducing the proportion of spermatozoa with damaged DNA.
Keywords: DNA fragmentation; Percoll density gradient; domestic horse; spermatozoa
How to cite
Gala K., Tarasek Z., Gurgul A., Kozlowska A., Bugno-Poniewierska M., 2026. Effectiveness of Percoll density gradient separation of Arabian stallion semen in selecting spermatozoa with preserved DNA integrity. In: 5th International PhD Students’ Conference at the University of Life Sciences in Lublin, Poland: Environment – Plant – Animal – Product. https://doi.org/10.24326/ICDSUPL5.A012