In Vitro and In Vivo Evaluation of the Fertilization Capacity of Frozen/Thawed Rooster Spermatozoa Supplemented with Different Concentrations of Trehalose
Status PubMed-not-MEDLINE Language English Country Switzerland Media electronic
Document type Journal Article
Grant support
MZE-RO0718
Ministry of Agriculture of the Czech Republic
SV23-7-21320
Ministry of Education Youth and Sports of the Czech Republic
PubMed
39765490
PubMed Central
PMC11672769
DOI
10.3390/ani14243586
PII: ani14243586
Knihovny.cz E-resources
- Keywords
- cryoprotectant, flow cytometry, gene reserve, sperm evaluation, trehalose,
- Publication type
- Journal Article MeSH
The objective of this study was to evaluate the impact of the supplementation of varying concentrations of the impermeable disaccharide trehalose on the in vitro and in vivo fertilization capacity of cryopreserved rooster spermatozoa in the original Czech Golden Spotted Hen breed. The control trehalose concentration was 0 mM, while TRE50 (50 mM), TRE100 (100 mM), and TRE200 (200 mM) were used as experimental trehalose concentrations. The kinematic and functional parameters of frozen/thawed spermatozoa were evaluated in vitro using mobile computer-assisted sperm analysis and a flow cytometer. The addition of 100 mM trehalose demonstrated the most favorable results for total (34.17%) and progressive (3.57%) motility after thawing. A statistically significant difference was found for these kinetic parameters compared to the other monitored concentrations. This experimental group was also found to have a significantly higher percentage of spermatozoa without plasma membrane or acrosome damage (33.37%) compared to the TRE50 group (30.74%; p < 0.05) and the TRE200 group (29.05%; p < 0.05). In vivo, artificial insemination was performed to verify fertilization ability. Hens (n = 40) were artificially inseminated twice (10 hens/treatment) with a 3-day interval between inseminations. In conclusion, the addition of 100 mM trehalose significantly improved total and progressive motility after thawing and preserved plasma membrane and acrosome integrity (p < 0.05). The fertilization rate of eggs fertilized with semen frozen with the addition of 100 mM trehalose was not significantly different from the other concentrations tested or the control group but was numerically higher (23.21% vs. 15.20% of fertilized eggs in this group).
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