2026-10-14T13:40:18Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33656032 2026-09-12T09:29:04Z category_24121 category_23902 item_type_3 month_year_09_2026

<b>Comparative sperm metabolomics reveals energy metabolism pathways associated with post-thaw sperm quality in indigenous Sumbawa and Warmblood stallions</b> Hikmayani Iskandar (22478027) Muhamad Aldi Nurdiansyah (22329466) Tulus Maulana (22478033) Ekayanti Mulyawati Kaiin (22478036) Syahruddin Said (22478030) Reski Amalia (22478057) Veterinary sciences not elsewhere classified Animal reproduction and breeding metabolomics cryopreservation spermatozoa stallion sperm <p dir="ltr"><b>Background:</b> Sperm cryopreservation can impair sperm function and produce variation in post-thaw sperm quality. Energy metabolism is important for sperm motility and membrane integrity, but metabolic signatures associated with post-thaw sperm quality differences between stallion populations remain poorly understood.</p><p dir="ltr"><b>Objectives:</b> To compare sperm metabolomic profiles of indigenous Sumbawa and Warmblood stallions and identify metabolic pathways and metabolites associated with post-thaw sperm quality.</p><p dir="ltr"><b>Methods:</b> Frozen semen from four healthy stallions, comprising two Sumbawa and two Warmblood stallions, was evaluated after thawing. Sperm quality was assessed using computer-assisted sperm analysis and hypo-osmotic swelling test, FITC-PNA, acridine orange, and CMA3 staining. Intracellular metabolites were analyzed using gas chromatography–mass spectrometry (GC–MS). Metabolic profiles were evaluated using hierarchical clustering and partial least squares discriminant analysis (PLS-DA) and KEGG pathway analysis.</p><p dir="ltr"><b>Results:</b> Sumbawa stallions exhibited higher curvilinear velocity, straight-line velocity, beat cross frequency, total motility, progressive motility, and plasma membrane integrity than Warmblood stallions. Sixteen metabolites were identified. Lactic acid, citric acid, and succinic acid. Lactic acid was more abundant in Sumbawa spermatozoa, whereas citric and succinic acids were more abundant in Warmblood spermatozoa. PLS-DA showed group separation (R² = 0.90, Q² = 0.73), supported by permutation testing. Glycolysis/gluconeogenesis, pyruvate metabolism, and the citrate cycle were differentiating pathways. Lactic acid was negatively associated with selected motility parameters, whereas citric and succinic acids were positively associated with selected sperm-quality parameters.</p><p dir="ltr"><b>Main Limitations:</b> The small number of stallions limits generalizability. Relative GC–MS abundance does not establish causality.</p><p dir="ltr"><b>Conclusions:</b> Sumbawa and Warmblood stallions exhibited distinct post-thaw sperm functional and metabolic profiles. Differences in key organic acids were associated with sperm motility and membrane integrity and linked to central energy metabolism. Sperm metabolomics may complement conventional sperm-quality assessment and identify candidate signatures associated with post-thaw function and potential cryotolerance. Larger studies are required for validation.</p> 2026-09-12T09:29:04Z Dataset Dataset 10.6084/m9.figshare.33656032.v1 https://figshare.com/articles/dataset/_b_Comparative_sperm_metabolomics_reveals_energy_metabolism_pathways_associated_with_post-thaw_sperm_quality_in_indigenous_Sumbawa_and_Warmblood_stallions_b_/33656032 CC BY 4.0