Effect of aminoguanidine supplementation on sperm cryopreservation in the domestic cat

Bc. Anna Sophová

Master's thesis

Abstract:

The effects of antioxidant supplementation of sperm samples intended for cool storage and cryopreservation have been investigated in multiple species during the past several decades. Various antioxidants have the ability to protect spermatozoa against oxidative stress by acting as free radical scavengers and oxidation inhibitors. The positive effect of aminoguanidine on sperm parameters such as motility and plasma membrane integrity has previously been documented in boar and rat. The present thesis is the first to evaluate the effect of aminoguanidine supplementation on sperm samples of the domestic cat (Felis catus). We hypothesized that supplementation of aminoguanidine would improve plasma membrane integrity, total and progressive motility and kinetic parameters in tomcat sperm samples subjected to cryopreservation and thawing. Sperm samples obtained from the epididymides after routine orchiectomies of 5 different domestic cats were supplemented with aminoguanidine in two concentrations- 1 mM and 5 mM, prior to cooling and cryopreservation. The samples were evaluated after 20- and 90- minutes of post-thaw incubation. The integrity of the sperm plasma membrane was evaluated with the use of a propidium iodide and carboxyfluorescein diacetate (PI/CFDA) staining solution. Total motility, progressive motility and kinetic parameters were evaluated using Computer Assisted Sperm Analysis (CASA). The age of the tomcats ranged from 9 to 11 months, the average body mass was 3,600 g. The freezing-thawing process negatively impacted all sperm parameters observed, which is in agreement with previous studies investigating the deleterious effects of cryopreservation. Progressive motility and plasma membrane integrity evaluated post-thaw showed higher values after supplementation of both concentrations of aminoguanidine. However, the difference between the CTR groups and the samples supplemented with aminoguanidine was not statistically significant. Supplementation of 1 mM and 5 mM of aminoguanidine did not have a significantly positive nor negative effect on total motility and kinetic parameters of tomcat sperm samples.

Keywords

antioxidants, free radicals, oxidative stress, sperm storage

Language used: English

Date on which the thesis was submitted / produced: 14. 4. 2022

Thesis defence

• Supervisor: Dott. Ric. Eliana Pintus

Czech University of Life Sciences Prague

Faculty of Agrobiology, Food and Natural Resources

Master programme / specializace:
Biotechnology / Reproduction Biotechnology