Effects of exercise and oral antioxidant supplementation enriched in (n-3) fatty acids on blood oxidant markers and erythrocyte membrane fluidity in horses.

The aim of this study was to investigate in a placebo-controlled field study the effect of a (n-3)-vitamin supplementation on erythrocyte membrane fluidity (EMF), oxidant/antioxidant markers and plasmatic omega3/omega6 fatty acid ratio (FAR) in 12 eventing horses. Venous blood was sampled at rest before (PRE) and after (POST) a three week treatment period with either the supplement (group S, n=6) or a placebo (group P, n=6) as well as after 15min (POST E15') and 24h (POST E24h) after a standardised exercise test. The following markers were analysed: EMF, plasma antioxidant capacity of water and lipid soluble components, ascorbic acid, uric acid (UA), glutathione (reduced: GSH, oxidised: GSSG), vitamin E (Vit E), beta-carotene, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, selenium, copper (Cu), zinc (Zn), oxidised proteins (Protox), lipid peroxides (Pool) and FAR. EMF did not differ between group S and P after treatment, but GPx remained unchanged in group S whereas it decreased in group P and plasma Cu/Zn ratio remained unchanged whereas it increased in group P. FAR were significantly increased in group S. Exercise induced a significant decrease of EMF (POST vs. E24h) in both groups, but which was significantly lower at E15' in group S than in group P. Exercise induced a significant increase of UA and ACW (POST vs. E15') and Protox (POST vs. E24h) in both groups. An exercise-related decrease in GSH and Pool (POST vs. E15') was found in group P, whereas Vit E and FAR (POST vs. E24h) significantly decreased in both groups. The study showed that exercise induced a decrease in EMF in horses associated with changes of blood oxidative balance. The (omega-3)-vitamin supplementation tested improved the oxidative balance poorly but delayed the exercise-induced decrease of EMF and increased the FAR.

Investigation of blood oxidant/antioxidant markers in healthy competition horses of different breeds.

REASONS FOR PERFORMING STUDY: There is increasing evidence that the equine athlete is exposed to exercise-induced changes of its oxidant/antioxidant balance and antioxidant supplementation is frequently recommended. However, it is unknown whether there is a specific need for antioxidants according to performance, breed, gender or age. OBJECTIVES: To assess whether breed-, gender- and age-related differences of blood oxidant/antioxidant markers occur in competition horses. METHODS: Healthy horses (n = 493) underwent oxidant/ antioxidant blood marker determination. Vitamin E, lipophilic antioxidant capacity (ACL), ascorbic acid (AA), glutathione (GSH, GSSG), gluthione peroxidase (GPx), superoxide dismutase (SOD), selenium (Se), copper (Cu), zinc (Zn), lipid peroxides (Pool), oxidised proteins (Protox) were determined, as well as magnesium (Mg), creatine phosphokinase (CPK), lactate dehydrogenase (LDH), packed cell volume (PCV) and haemoglobin (Hb). A mixed linear model assessed the effect of breed, gender and age category. P<0.05 was considered significant. RESULTS: Thoroughbreds showed the highest values of vitamin E, ACL, GPx, PCV and Hb, whilst standardbreds had the highest values of AA and LDH. Jumping horses had the highest Protox values. Females had significantly higher SOD values, whereas most of the other markers were higher in stallions and geldings. Horses age 2-6 years had higher AA, SOD and LDH values than horses age >6 years. Correlation analyses were positive and significant between vitamin E and GPx, VitE and ACL, Se and GPx, Cu and Pool and negative between Pool and vitamin E, Pool and ACL, Protox and GPx, Protox and vitamin E. CONCLUSIONS: Blood oxidant/ antioxidant status of horses is influenced by breed, gender and age. The correlation analyses suggest synergistic relations between GPx, vitamin E and Se and an antagonistic relation between Protox-GPx, Protox-vitamin E, and Pool-vitamin E. POTENTIAL RELEVANCE: The results of this investigation provide definition of the specific need for antioxidants and vitamins in competition horses.

Effect of exercise on blood oxidant/antioxidant markers in standardbred horses: comparison between treadmill and race track tests.

REASONS FOR PERFORMING STUDY: Interest in establishing oxidant/ antioxidant profiles in competition horses is increasing. Earlier studies performed in horses have mainly been performed under laboratory conditions using a treadmill and it is not known to what extent laboratory results of oxidant/antioxidant studies might be transposed to field conditions. OBJECTIVE: To compare the impact on the blood oxidant/ antioxidant status of a standardised exercise test including a run up to fatigue performed on a treadmill (TM) and on a racetrack (RT) in healthy and trained Standardbred horses. MATERIAL AND METHODS: During TM and RT tests the following blood antioxidant markers were analysed in jugular venous blood at rest and 15 mins (E15) after an intense bout of exercise: uric acid (UA), ascorbic acid (AA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (reduced: GSH and oxidised: GSSG), glutathione redox ratio (GRR) and protein thiol (PSH). Running time to fatigue (RTF), velocity during the last exercise stage (Vmax), final heart rate (HRfinal) and venous lactic acid (LA) were also recorded. RESULTS: Vmax was significantly (P<0.05) higher during the RT, whereas LA was significantly lower. HRfinal and RTF did not differ significantly between TM and RT. Exercise induced a significant increase (R vs. E15) of UA and AA in both tests, whereas GSH and PSH decreased significantly. GPx, SOD, GSSG and GRR remained unchanged. Differences between TM and RT were significant at E15 for UA, AA and PSH. CONCLUSIONS: Comparison of oxidant/antioxidant profiles from laboratory and field studies are difficult to standardise and should be interpreted with caution. POTENTIAL RELEVANCE: For the same RTF and final HR, the TM induced stronger changes in blood lactate and in blood oxidant/antioxidant balance than did RT.

The effects of dietary N-3 and antioxidant supplementation on erythrocyte membrane fatty acid composition and fluidity in exercising horses.

REASONS FOR PERFORMING STUDY: Fatty acid supplementation could modulate erythrocyte membrane fluidity in horses at rest and during exercise, but information is lacking on the effect of exercise. OBJECTIVES: To assess the effect of exercise with, and without, an oral antioxidant supplementation enriched with n-3 fatty acids on erythrocyte membrane fluidity (EMF) and fatty acid composition in eventing horses. METHODS: Twelve healthy and regularly trained horses were divided randomly into 2 groups: group S received an oral antioxidant cocktail enriched in n-3 fatty acid (alphatocopherol, eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) whereas group P was placebo-treated. At the end of 4 weeks, all horses performed a standardised exercise test (ET) under field conditions. Venous blood was sampled before starting treatment (TO), immediately before (T1) as well as 15 min (T2) and 24 h (T3) after ET. Spin labelled (16-DOXYL-stearic acid) red blood cell membranes were characterised using the relaxation correlation time (Tc in inverse proportion to EMF). Fatty acid composition (%) of the membrane was determined by gas-liquid chromatography. RESULTS: Supplementation did not induce changes in EMF (T1 vs. TO) but significant changes in membrane composition were observed and there were increases in n-3 polyunsaturated fatty acid PUFA, n-3/n-6 ratio, and total n-3 fatty acids. Exercise (T2 vs. T1) induced a significant decrease of EMF in group P (Tc: +19%, P<0.05) and nonsignificant decrease in group S (Tc: +5%), whereas membrane fatty acid composition did not change in either group. During the recovery period (T3 vs. T2), EMF decreased significantly in group S (Tc: +29%, P<0.05) and nonsignificantly in group P (Tc: +18%) without any significant changes in fatty acid composition. CONCLUSION AND POTENTIAL RELEVANCE: An enriched oral antioxidant supplementation induced changes in membrane composition, which modulated the decrease in EMF induced by exercise. Long chain n-3 fatty acid supplementation might therefore be beneficial.

Validation of a portable equine metabolic measurement system.

REASONS FOR PERFORMING STUDY: In equine sports medicine, VO2 has been measured exclusively with stationary systems, in laboratories equipped with a treadmill. Measurement during exercise in field conditions has not previously been reported because of the lack of portable equipment designed for horses. OBJECTIVES: A commercially available portable metabolic measurement system, based on breath-to-breath gas analysis and flow spirometry, was adapted to the horse's physiology and morphology (Cosmed K4b2 and Equimask) and its validity tested by (1) repeatability of the measures and (2) comparing metabolic data to those obtained by a reference method (RM). METHODS: To test the reproducibility of the measurements, 5 healthy saddle horses were subjected twice at 2 day intervals to a similar submaximal standardised incremental exercise test on a treadmill. The same horses performed twice at one week interval an incremental treadmill test to fatigue: the oxygen consumption and ventilation were measured once with the K4b2 system and once with the RM. The metabolic and ventilatory data obtained with both systems were compared. RESULTS: There was a good reproducibility of the metabolic measurements obtained by the K4b2 system at any workload. The VO2 obtained by both systems at any workload was not significantly different. However, the K4b2 expired fraction in CO2 (FETCO2) and carbon dioxide production (VCO2) were significantly lower at high and at maximal workloads. As a consequence, the values of the respiratory exchange ratio were too low and incompatible with normal physiological values. CONCLUSIONS: The good reproducibility of the metabolic and ventilatory measurements and the fact that the VO2 measurements at any workload were similar to the data obtained with the reference method suggested that this system may be used for comparison of repeated VO2 measurements in practical field conditions. POTENTIAL RELEVANCE: The K4b2 system could be used to improve knowledge of the energetic cost in different equine sports disciplines and offer the opportunity to undertake performance tests with genuine track conditions, on ridden or harnessed horses, rather than under laboratory conditions.

Effect of oral antioxidant supplementation on blood antioxidant status in trained thoroughbred horses.

The oxidant/antioxidant equilibrium of trained thoroughbred horses (n = 40) was assessed on three occasions during a period of three months under field conditions by blood antioxidant markers analysis, i.e. plasma ascorbic acid (AA), plasma antioxidant capacity of water-soluble components (ACW), whole blood (GSH) and oxidised (GSSG) glutathione, plasma alpha-tocopherol and beta-carotene, plasma antioxidant capacity of lipid-soluble components (ACL), red blood cell superoxide dismutase (SOD) and glutathione-peroxidase activity (GPx) and plasma trace-elements, i.e. selenium (Se), copper (Cu), zinc (Zn). A control group of ten horses receiving a placebo and an antioxidant group of 30 horses orally supplemented with an antioxidant mixture were randomly formed. An antioxidant imbalance was observed after three months in the control group, reflected by a significant decrease in GSH, SOD, GPx, Se (P < 0.05) and a significant increase in GSSG (P < 0.05). The antioxidant supplement prevented GPx and Se decrease and significantly increased ACW, alpha-tocopherol, beta-carotene and ACL (P < 0.05). Significant sex- or age-related differences were found for AA, ACW, alpha-tocopherol, SOD, GPx and Se, and there were significant correlations between ACW-AA, ACL-alpha-tocopherol, GPx-Se, CPK-Se, CPK-alpha-tocopherol and CPK-Cu. This field study has shown that trained thoroughbred horses undergo significant changes of several blood antioxidant markers and that oral antioxidant supplementation might partially counterbalance these changes by improving the hydrophilic, lipophilic and enzymatic antioxidant blood capacity.

Relationship between markers of blood oxidant status and physiological variables in healthy and heaves-affected horses after exercise.

Exercise-induced oxidative stress is investigated as a potential performance-limiting factor in human sports medicine. Therefore, the present study aimed to assess whether physiological variables that change with exercise intensity were correlated with blood oxidant markers in healthy and heaves-affected horses. Seven healthy horses, 8 heaves-affected in remission and 7 heaves-affected in crisis performed a standardised exercise test (SET) of stepwise increasing intensity. Variables monitored during exercise were heart rate (HR), venous plasma lactate (LA), packed cell volume (PCV) and arterial oxygen tension (PaO2). Oxidant markers (uric acid [UA], 8-iso-PGF2alpha and reduced [GSH] and oxidised glutathione [GSSG]) were analysed in venous peripheral blood sampled at rest (R), at peak-exercise intensity (Emax), 15 (E15) and 60 (E60) min after SET. There was a significant effect of heaves on oxidant markers and, therefore, correlation analyses between physiological variables and oxidant markers were performed separately per horse group. In healthy horses, UA analysed at Emax was positively correlated with LA. Furthermore, GSH analysed at Emax and E15 was positively correlated with PaO2. In healthy and heaves-affected horses in remission, GSH and GSSG determined at Emax were negatively correlated with HR. There was no significant correlation between 8-iso-PGF2alpha and physiological variables. In conclusion, a correlation between the physiological response to exercise and some oxidant markers exists in healthy horses. However, in heaves-affected horses the blood oxidant status is probably more dependant on airway disease than on exercise. Future studies should be undertaken to assess whether antioxidant supplementation might positively influence the oxidant-antiodidant balance in exercising horses.