Relationships between Seminal Plasma Metabolites, Semen Characteristics and Sperm Kinetics in Donkey (Equus asinus)

SIMPLE SUMMARY: A deeper knowledge of reproductive biology may be helpful in the donkey to avoid the risk of extinction that some breeds are facing. The evaluation of metabolites in seminal plasma provides crucial information for the knowledge of donkey sperm metabolism, for obtaining comparative information with other species, as well as for providing useful elements for the formulation of extenders for sperm dilution and conservation. Moreover, correlations of seminal metabolites with sperm kinetics highlight new possible markers of sperm quality. Using multivariate analysis, all metabolic, seminal, and spermatic data were merged in a single dot that grouped individual stallions within clusters in the Cartesian axes according to the different spermatic characteristics. This amount of information also allows to shed light on the effects of total or partial removal of seminal plasma for improving sperm preservation. The inclusion in the study of an azoospermic individual represents a further discriminating element in the analysis of sperm quality under physiological and pathological conditions. ABSTRACT: This study aimed to evaluate donkey seminal plasma metabolites and relate this information to the main characteristics of sperm quality. Sperm kinetics from 10 donkey stallions were analyzed with a computerized system at the time of collection (T0) and after 24 h storage at 4 °C (T24). Seminal plasma was frozen at −80 °C for subsequent proton nuclear magnetic resonance ((1)H NMR) spectroscopy. On three stallions, semen collection was repeated monthly for three times and sperm analysis also included mitochondrial activity and oxidative status. One stallion was azoospermic and a second semen collection was performed after one month. In the seminal plasma, 17 metabolites were identified; their levels showed numerous significant variations between the azoospermic and the normospermic individuals and grouped in well-defined clusters in a multivariate analysis. Comparing individuals with high and low sperm motility, the only discriminating metabolite was phenylalanine, whose levels were lower in the latter, as in the azoospermic individual. Phenylalanine was also the only metabolite highly correlated with all sperm kinematic parameters at T24. In conclusion, the present study has provided relevant information on the chemical characteristics of donkey semen, identified relationships between seminal metabolites, semen parameters, and sperm kinetics, and offered insights for future technological applications.