Acta Vet. Brno 2023, 92: 21-26

Effect of sperm and glycerol concentration on epididymal sperm motility parameters after thawing

Petra Andrlová1, Miroslava Mráčková1, Eva Jánová2,3, Filip Kološ1, Markéta Sedlinská1

1University of Veterinary Sciences Brno, Faculty of Veterinary Medicine, Equine Clinic, Brno, Czech Republic
2Ceitec VETUNI, Brno, Czech Republic
3University of Veterinary Sciences Brno, Faculty of Veterinary Medicine, Institute of Genetics, Brno, Czech Republic

Received July 12, 2022
Accepted December 13, 2022

The aim of this study was to evaluate the effect of different glycerol concentrations and different sperm concentrations on stallion epididymal sperm motility indicators after thawing. For statistical analysis, 25 stallions were used. Collection of the epidydimal spermatozoa was performed as a retrograde flush of cauda epididymis and ductus deferens within 12 h post castration. After evaluation, the resuspended spermatozoa were centrifuged, the supernatant removed and the spermatozoa resuspended in Gent semen extender to get three different groups with different concentrations of sperm (250 × 106 in ml, 500 × 106 in ml, 1000 × 106 in ml) and different final glycerol concentrations (2%, 4%, and 6%). Therefore, 9 different samples were finally obtained from each stallion. The spermatozoa were packed and placed in a fridge (4 °C) for 2 h, then placed in liquid nitrogen vapour (−80 to −100 °C) and after 25 min plunged into the liquid nitrogen and stored at −196 °C for at least 5 days. The selected straws were individually thawed in a 38 °C water bath for 30 s prior to post-freezing analysis. Motility indicators were assessed at 0, 60, 120, and 180 min after thawing. Parametric test was used for analysis; the measured indicators were total motility, progressive motility, curvilinear velocity, straightness, and average-path velocity. In this study, the best results were reached in samples diluted into a concentration of 1,000 × 106 in ml, regardless of the concentrations of glycerol.


This project was supported by the Internal Grant Agency of the University of Veterinary Sciences (IGA 115/2020/FVL and IGA 111/2021/FVL).


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