Plasma amino acid (PAA) concentration in horses vary according to the exercise type. This study evaluated the changes in PAA levels and the associated metabolites, urea and ammonia, following short-duration, high-intensity cross-country exercise in eventing horses. Twenty eventing horses participated in 55 rides at 14 international competitions (2* to 4* levels) across five venues in Germany and Poland. Blood samples were collected at four timepoints: before exercise (TP0), at 10 min (TP1), and at 30 min (TP2) post-exercise, as well as in the morning on the day after the competition (TP3). A total of 23 different PAAs and two metabolites (ammonia and urea) were analyzed. PAA concentration difference over time was assessed by a mixed ANOVA. Significant fluctuations were observed in 18/25 parameters. For 21/23 PAAs, levels increased at TP1 and/or TP2, while cysteine concentrations decreased. Concentrations returned to pre-competition levels for 21/23 PAAs by TP3. Proline levels remained elevated (p = 0.002), while those of glycine significantly decreased (p = 0.027) at TP3. Plasma ammonia and urea levels increased at TP1, TP2 and TP3. This study provides foundations for supplementation strategies and can inform future works exploring PAAs’ role in performance and training adaptation in eventing horses and their potential as performance-related biomarkers.

Amino acid (AA) metabolism and the adaptation of muscles to increased physical performance are interdependent. This field study examined changes in plasma amino acid (PAA) concentrations and two metabolites (ammonia and urea) before and after cross-country (CC) competitions throughout an entire eventing season and their associations with performance outcomes in eventing horses. Twenty horses from nine different Warmblood breeds were evaluated between two and seven times across 14 international eventing competitions over a 23-week period. A total of 55 rides were sampled. Blood samples were collected at four time points: before the start (TP0), 10 min after completion of the CC ride (TP1), 30 min post-CC (TP2), and the next morning (TP3). A mixed ANOVA revealed significant effects (p < 0.05) of calendar week on PAA concentrations for asparagine, ornithine, and proline. Correlations between CC scores and PAA concentrations were observed for 4 of the 25 measured parameters. TP0 leucine concentrations and TP2 histidine concentrations correlated negatively with CC penalty score, whereas TP1 proline concentrations and TP3 alanine concentrations correlated positively with CC penalty score. These results indicate that higher plasma leucine concentrations may contribute to improved performance in eventing horses. Additionally, alanine and proline show potential as performance-related diagnostic biomarkers.