Volume: 5, 2026
5th International PhD Students’ Conference at the University of Life Sciences in Lublin, Poland:
ENVIRONMENT – PLANT – ANIMAL – PRODUCT
Abstract number: A033
DOI: https://doi.org/10.24326/ICDSUPL5.A033
Published online: 22 April 2026
Laboratory analysis of ram sperm quality
Filipp Georgijevič Savvulidi*, Martin Ptáček and Lucie Langerová
Department of Animal Science, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences (CULS) Prague, Czech Republic
* Corresponding author: savvulidi@af.czu.cz
Recent advances in laboratory diagnostics have substantially improved the assessment of livestock sperm quality, with particular relevance to ovine reproduction. Ram spermatozoa are highly susceptible to cryodamage, making the evaluation of frozen–thawed semen quality a critical step prior to artificial insemination. Reliable in vitro assessment of semen doses supports informed decisions on insemination methodology (cervical versus laparoscopic), optimization of insemination strategies (e.g., number of doses per ewe, single or double insemination), and prediction of reproductive outcomes.
A comprehensive evaluation of sperm quality includes key parameters such as motility, as well as structural and functional integrity of cellular organelles. Among available analytical tools, computer-assisted sperm analysis (CASA) represents a highly informative and powerful technique for motility assessment, enabling detailed quantification of parameters such as velocity and progressive movement. Furthermore, advanced data processing approaches, including clustering analysis of CASA outputs, provide deeper insight into sperm subpopulations and functional heterogeneity.
Flow cytometry has emerged as another essential tool, allowing high-throughput evaluation of sperm viability and functionality using a range of fluorescent probes, from classical dyes such as propidium iodide to newer alternatives like aniline blue. However, flow cytometry presents limitations, notably the lack of direct visualization of fluorescence localization on sperm cells. This can complicate interpretation in certain assays, such as those assessing oxidative stress using CellROX dyes. Therefore, validation of flow cytometry results through complementary techniques, including fluorescence microscopy or imaging flow cytometry, is strongly recommended.
Functional assays, particularly the hypoosmotic swelling test, remain indispensable for evaluating sperm membrane integrity. Despite the value of laboratory analyses, these methods provide only preliminary estimates of fertilizing potential. Consequently, in vitro findings must be validated through field fertility trials.
Additionally, predicting ejaculate freezability is of practical importance, and laboratory tests assessing sperm tolerance to dilution may offer useful indicators.
Overall, integrating advanced laboratory diagnostics with field validation enhances the reliability of reproductive management in sheep.
Keywords: artificial insemination, CASA, cryodamage, flow cytometry, ram sperm
How to cite
Savvulidi F.G., Ptáček M., Langerová L., 2026. Laboratory analysis of ram sperm quality. 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.A033