Horses competing in cross-country tests are subjected to high physical demands. Within the scope of this prospective longitudinal study, blood values of 20 elite eventing horses were examined before and after two- to four-star cross-country rides. The aim was to find out whether blood-based markers for skeletal muscle and cardiac muscle function change after cross-country exercise. Parameters that provide information about fluid balance, muscle enzymes, metabolites and cardiac muscle-specific markers were investigated. We developed an approach to eliminate the concentration changes caused by reduced plasma volume. Parameters were measured pre, 10 and 30 min post exercise and the next morning and were evaluated using a mixed model. Thirty minutes after exercise, most parameter concentrations changed in an exercise-dependent manner. The next morning, most exercise-related markers recovered rapidly, while creatine kinase (CK) (26% increase; p = 0.008) and lactate dehydrogenase (LDH) (15% increase; p < 0.001) showed a declining but sustained increase. Cardiac troponin I (cTnI) increased above the reference range in 40 of the 55 rides (73%) and in 18 of 20 horses in the morning after exercise.

Background: After submaximal exercise, blood values of eventing horses show physiological reactions.Objectives: This prospective longitudinal study investigated blood parameters in 20 elite eventing horses before and after two–four‐star cross‐country rides.Methods: Using a mixed model adjusting for plasma volume shift, we assessed exercise‐dependent parameters and compared blood values with reference ranges for healthy horses at rest.Results: Following exercise, cortisol, triiodothyronine (T3) and thyroxine (T4) showed short‐term increases, and superoxide‐dismutase showed a small short‐term increase. Hepatic values showed short‐term (haemoglobin [HGB], globulins) or sustained increases (bilirubin, glutamate dehydrogenase, alanine aminotransferase). Digestion‐related parameters showed small short‐term increases (α‐amylase, triglycerides) or decreases (cholesterol, DGGR‐lipase), apparent through plasma shift adjustment. Zinc decreased in the short term, and iron showed a delayed decrease. White blood cell count increased persistently after training, whereas serum amyloid A remained unchanged.Conclusions: Exercised eventing horses had consistently elevated HGB and cortisol levels 10 and 30 min after submaximal exercise, exceeding the reference ranges for healthy horses at rest. Exercise activates the hypothalamic–pituitary–adrenocortical and hypothalamic–pituitary–thyroid axes. Antioxidant activity was observed. Increased energy requirements led to the mobilization of energy reserves, and a sustained increase in liver enzymes indicated hepatocellular injury. Mild haemolysis suggested increased muscle metabolism, whereas signs of inflammation were subtle. Further research is needed to identify which horses deviate from mean values.

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.

This study examines seasonal changes in muscle and heart parameters in eventing horses over the course of a competition season. Blood levels of the enzymes creatine kinase (CK) and aspartate aminotransferase (AST) as well as the heart muscle-specific concentration of cardiac troponin I (cTnI) were measured before (pre), 30 min (p30) and 24 h (p24) after competitions. Creatine kinase (CK: median pre-competition = 175 U/L, 30 min post = 221 U/L, 24 h post = 140 U/L), aspartate aminotransferase (AST: pre = 319 U/L, p30 = 335 U/L, p24 = 333 U/L), and cardiac troponin I concentration (cTnI: pre = 0.006 ng/mL, p30 = 0.011 ng/mL, p24 = 0.007 ng/mL) exhibited partial normalization by 24 h post-exercise but at the same time demonstrated significant seasonal variation (p < 0.001). Echocardiographic assessments revealed sustained high-level myocardial velocities, with occasional modest seasonal declines. A significant correlation was identified between cTnI levels and early diastolic myocardial velocity (Em) (Spearman’s Rho: pre-exercise 0.323, Rho p30: 0.357), whereas a negative correlation was manifest at 24 h (Spearman’s Rho = −0.300). These findings suggest a heightened sensitivity of diastolic myocardial velocity to myocardial injury. Given that diastolic dysfunction frequently constitutes an early manifestation of myocardial compromise, our results underscore the utility of biomarkers alongside myocardial velocity measures as valuable tools for the early detection of subclinical fatigue in high-performance sport horses.