Acta Vet. Brno 2024, 93: 59-64

Sperm storage of wild cervid species: comparison of available cryoprotectants

Vladimír Piaček1, Jan Zukal2, Veronika Seidlová1, František Vitula1, Šárka Bednaříková1, Heliana Dundarova3, Barbora Havelková1, Monika Němcová1, Kateřina Zukalová1, Jiří Pikula1

1University of Veterinary Sciences Brno, Faculty of Veterinary Hygiene and Ecology, Department of Ecology and Diseases of Zoo Animals, Game, Fish and Bees, Brno, Czech Republic
2Academy of Sciences of the Czech Republic, Institute of Vertebrate Biology, Brno, Czech Republic
3Bulgarian Academy of Sciences, Institute of Biodiversity and Ecosystem Research, Sofia, Bulgaria

Received April 26, 2023
Accepted February 19, 2024

We compared four diluents (Andromed, dimethylacetamide, dimethyl sulphoxide, Triladyl) used for semen cryopreservation in wild ungulates from the family Cervidae, i.e. roe deer (Capreolus capreolus), red deer (Cervus elaphus), and fallow deer (Dama dama). Epididymal sperm samples were collected and analysed immediately using Computer Assisted Sperm Analysis with the module for concentration and motility determination and, after equilibration, frozen and placed in liquid nitrogen for one month. After thawing, the samples were again subjected to the same examination procedure. Influence of cryopreservation and choice of the cryoprotectant was assessed by monitoring sperm concentration, average head area (HA), average velocity and progressivity (VAP), beat frequency (BF), total motility (MO), total progressive motility (PR), circular tracks (CT) and mucous penetration (MP). We observed no significant differences in sperm concentration between cryoprotective diluents or fresh and post-thaw samples in all species. All motility indicators (MO, PR, VAP, BF, CT) were influenced by the treatment but did not differ significantly between diluents used in red and roe deer. In fallow deer, commercial diluents (Andromed, Triladyl) resulted in better sperm survival than the alternatives (dimethyl sulphoxide, dimethylacetamide). Only HA showed significant differences (P < 0.001) in all species based on the diluent, with no effect of treatment. In contrast, MP was influenced by both the diluent and the cryopreservation process in roe deer and, partly, fallow deer. In future studies, we suggest expanding both the members of the Cervidae family examined and the sample size. Knowledge how to optimise cryopreservation protocols for different mammalian species has implications for conservation reproductive medicine of endangered wildlife.


This research was supported through project FVHE_Pikula_2023ITA25. The funders had no role in the study design, data analysis, decision to publish or preparation of the manuscript.


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