Effects of exercise intensity and duration on plasma beta-endorphin concentrations in horses

OBJECTIVE: To determine the relationship between plasma beta-endorphin (EN) concentrations and exercise intensity and duration in horses. ANIMALS: 8 mares with a mean age of 6 years (range, 3 to 13 years) and mean body weight of 450 kg. PROCEDURE: Horses were exercised for 20 minutes at 60% of maximal oxygen consumption (VO2max) and to fatigue at 95% V02max. Plasma EN concentrations were determined before exercise, after a 10-minute warmup period, after 5, 10, 15, and 20 minutes at 60% VO2max or at the point of fatigue (95% VO2max), and at regular intervals after exercise. Glucose concentrations were determined at the same times EN concentrations were measured. Plasma lactate concentration was measured 5 minutes after exercise. RESULTS: Maximum EN values were recorded 0 to 45 minutes after horses completed each test. Significant time and intensity effects on EN concentrations were detected. Concentrations were significantly higher following exercise at 95% VO2max, compared with those after 20 minutes of exercise at 60% VO2max (605.2 +/- 140.6 vs 312.3 +/- 53.1 pg/ml). Plasma EN concentration was not related to lactate concentration and was significantly but weakly correlated with glucose concentration for exercise at both intensities (r = 0.21 and 0.30 for 60 and 95% VO2max, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: A critical exercise threshold exists for EN concentration in horses, which is 60% VO2max or less and is related to exercise intensity and duration. Even under conditions of controlled exercise there may be considerable differences in EN concentrations between horses. This makes the value of comparing horses on the basis of their EN concentration questionable.     

Effects of oral cimetidine on plasma concentrations of phenylbutazone in horses

Phenylbutazone was administered to six Thoroughbred horses in a cross-over study in which the horses received cimetidine pretreatment or no cimetidine pretreatment. Blood samples were collected at various times for 48 h after phenylbutazone administration and the plasma was analysed for phenylbutazone. Cimetidine pretreatment elevated phenylbutazone plasma concentrations during the first 8 h after phenylbutazone administration. The absorption rate, maximum phenylbutazone plasma concentrations and AUC were significantly greater with cimetidine pretreatment. The half-life of phenylbutazone did not change with cimetidine pretreatment; however, lower plasma concentrations of the metabolite gamma-hydroxyphenylbutazone were observed with cimetidine pretreatments. Plasma concentrations of the metabolite oxyphenbutazone were unchanged with cimetidine pretreatment compared to control values. Twenty-four-hour plasma concentrations of phenylbutazone were not different from control values with cimetidine pretreatment. This study suggests that concurrent treatment with cimetidine and phenylbutazone 24 h before race time does not result in elevations of plasma phenylbutazone concentrations above control values.     

Plasma potassium and lactate concentrations in thoroughbred horses during exercise of varying intensity.

To investigate the effect of moderate to high intensity exercise of up to 6 min duration on plasma potassium and lactate concentrations, 6 Thoroughbred horses were studied using a treadmill at a 5 degree incline. Each test consisted of an 8-min standardised warm-up followed by an exercise bout at 8, 9, 10 or 12 m/sec. The horses were galloped at each speed for up to a maximum of 6 min or until signs of fatigue were present. The horses were then walked at 0 degree incline. Carotid arterial blood samples were taken during and after the exercise. At 8, 9 and 10 m/sec there was a general pattern of an initial rise in potassium to a peak around 1.5 min of exercise with the concentration then slowly decreasing. At 12 m/sec there was a continuous rise to a peak at the end of exercise in all horses. Immediately after exercise there was a rapid return (within 3-4 min) to the potassium concentrations recorded at the end of the warm-up period. Plasma lactate peaked around the end of exercise at all speeds. At the highest intensity of exercise the mechanisms for the re-uptake of potassium did not appear to be able to match the rate of efflux. In contrast, at less intense work loads, the rate of re-uptake appeared to be similar to or slightly greater than the rate of efflux. It is possible that a disturbance in this balance between efflux and re-uptake could result in a disturbance in normal neuromuscular function during exercise.     

Effects of dietary cation-anion balance on acid-base status in horses

Four mature, sedentary geldings were used in a 4x4 Latin square design experiment to study the effects of dietary cationanion balance (DCAB), defined as meq ((Na + K) - C1)/kg of dietary dry matter, on urine pH, blood pH and blood gasses. Four diets with actual DCAB levels of +21 (Low, L), +125 (Medium Low, ML), +231 (Medium High, MH) and +350 (High, H) were fed for a 21-day adjustment period and a 72 hour collection period. Total urine output was collected every 4 hours post feeding for 72 hours, and like times across days were combined for analyses. Arterial and venous blood samples were drawn, via indwelling catheters, hourly for 12 hours beginning at feeding. All samples were analyzed immediately after collection for pH, blood gasses and blood acid-base measures. Urine pH was lower (p<.05) at all measured intervals when geldings consumed diet L than when they consumed diets MI, MH and H. Least squares means for urine pH ranges from 5.40 to 5.86 on diet L, 6.79 to 7.30 on diet ML, 7.35 to 7.63 on diet MH and 7.52 to 8.14 on diet H. Blood pH, pCO₂ and HCO₃ values were also significantly lower in both arterial and venous samples when geldings consumed diet L than when fed diets MH and H. Results of this trial indicate that horses consuming highly anionic diets may experience a nutritionally-induced metabolic acidosis.     

Blood-gas, acid-base and haematological values in horses during an endurance ride.

The effects of prolonged strenuous exercise on arterial and venous oxygen tension, carbon dioxide tension, pH, bicarbonate, standard bicarbonate, base excess, haemoglobin, packed cell volume and total plasma protein were studied in 36 horses during a 100 km endurance ride. There were significant changes in many parameters when pre-ride values were compared with both mid-ride and end of ride values. The prominent changes were the development of dehydration and a metabolic alkalosis. At the mid-ride sampling time those horses with higher heart rates had a greater degree of metabolic alkalosis than those with lower heart rates. The first 4 horses in the race completed the ride with speeds between 322-330 m/min and demonstrated a metabolic acidosis.     

Metabolic responses to oral tryptophan supplementation before exercise in horses

This study was conducted to evaluate the effects of oral tryptophan (Trp) supplementation on exercise capacity and metabolic responses in horses. Three horses had to perform an exercise test: a 15-min warm-up followed by a 60-min walk (1.7 m/s, W1), a 10-min trot (3.1 m/s, T1), a second 60-min walk (1.7 m/s, W2), a second 10-min trot (3.1 m/s, T2) and a final 30-min walk (1.7 m/s, W3) until the horses were unwilling to continue. The horses exercised on a treadmill at a 6% incline and with a constant draught load of 40 kg (0.44 kN). Two hours before exercise horses were given 50 g Trp (9.8-10.7 g Trp/100 kg BW) by nasogastric tube. A control exercise test was conducted without Trp. During the control test, one horse was able to finish the final 30-min walk (W3), whereas two horses finished W3 after Trp administration. Higher plasma Trp levels after Trp administration did not change significantly during exercise (Trp: start exercise, 524 +/- 41 micromol/l; end exercise 547 +/- 20 micromol/l; control: start exercise, 70 +/- 10 micromol/l; end exercise, 58 +/- 21 micromol/l). After Trp supplementation, blood lactate concentrations were significantly lower after the first and second trotting periods. Free fatty acids in plasma increased during exercise without any treatment-related differences. Although experimental plasma Trp levels were seven times higher than the control levels, Trp supplementation had no effect on exercise performance and metabolic responses to draught load exercise.     

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.     

Antioxidant supplementation and subsequent oxidative stress of horses during an 80-km endurance race

This study tested the development of oxidative stress and the effects of antioxidant supplementation in an 80-km ride. A precompetition survey revealed that no competitor would participate without vitamin E supplementation; therefore, 46 horses were paired for past performances and randomly assigned to two groups of 23 each for 3 wk of supplementation before the ride. One group (E) was orally supplemented with 5,000 IU of vitamin E per day; the other group (E+C) received that dose of vitamin E plus 7 g/d of vitamin C. Blood samples, temperature, and heart rate were taken the day before the race, at 21 and 56 km during the ride, at completion, and after 20 min of recovery. Plasma was assayed for lipid hydroperoxides, alpha-tocopherol, total ascorbate, albumin, creatine kinase (CK), and aspartate aminotransferase (AST). Total glutathione and glutathione peroxidase activity were determined in red blood cells and white blood cells. Thirty-four horses completed the race, 12 horses (six in E and six in E+C) did not finish for reasons including lameness, metabolic problems, and rider option. Plasma ascorbate was higher (P = 0.045) in the E+C group than in the E group. Other than ascorbate, neither antioxidant status nor CK and AST activities were affected by supplementation with E+C vs. E. Red blood cell glutathione peroxidase, white blood cell total glutathione, lipid hydroperoxides, CK, and AST increased, and red blood cell total glutathione and white blood cell glutathione peroxidase activity decreased with distance (P < 0.001). Positive correlations were found for plasma lipid hydroperoxides on CK (r = 0.25; P = 0.001) and AST (r = 0.33; P < 0.001). These results establish an association between muscle leakage and a cumulative index of oxidative stress.     

Clenbuterol plasma concentrations after repeated oral administration and its effects on cardio-respiratory and blood lactate responses to exercise in healthy Standardbred horses

To evaluate the effects of clenbuterol on cardio-respiratory parameters and blood lactate relation to exercise tolerance, experimental horses performed standardized exercise tests on a high-speed treadmill before and after administration of the drug. Clenbuterol was administered in feed to six healthy Standardbreds at a dose rate of 0.8 micrograms/kg b.wt twice daily for 5.5 days. Each horse was tested twice, without and with a respiratory mask, during two consecutive days. One week elapsed between the baseline tests without drug and the tests with clenbuterol treatment (each horse served as its own control). The results show an unchanged heart rate response to exercise 2 h after the last clenbuterol administration. The blood lactate response and the arterial oxygen tension during exercise did not differ before and after drug treatment. The oxygen uptake as well as pulmonary ventilation relative to the work load performed was essentially unaffected. The arterial pH during exercise was significantly increased (P less than 0.05) following clenbuterol treatment. Plasma levels of clenbuterol were maximal 2 h post-administration with values between 0.45 and 0.75 ng/ml. The plasma half-life of elimination was 10.4 h (+/- 2.25 SD). In conclusion, clenbuterol did not cause any major effects on the cardio-respiratory and blood lactate parameters studied in healthy horses performing submaximal exercise tolerance tests.     

Effects of selenium supplementation on plasma progesterone concentrations in pregnant heifers

It is known that selenium (Se) has various functions in animals. Many investigations on the biochemical and physiological effects of Se have been previously reported; however, the detailed function of Se in reproduction is not yet clear. We proposed the possibility that Se plays a notable role in progesterone production. The aim of this study was to clarify the effects of Se supplementation on progesterone levels of pregnant Holstein heifers. Eight Holstein heifers (?Se) were fed basal diet (containing 0.022 ppm of Se) throughout the experiment. While a 0.3 ppm diet of Se (sodium selenite) was fed to another seven animals (+Se) with basal diet. Blood sampling was carried out every week. Plasma Se concentrations were higher in Se‐supplemented cows compared with controls (?Se) (P < 0.01) throughout the experiment. Se supplementation increased plasma progesterone in the 29–39 weeks of pregnancy from 4.98 ± 0.64 to 6.86 ± 0.49 ng/mL on average (P < 0.05). The present findings suggest that Se contributes to maintaining the function of the corpus luteum and/or placenta in the latter period of pregnancy.     

Growth hormone secretion in relation to plasma lactate and glucose concentrations during a maximal treadmill exercise test in horses

Veterinary Research Communications -     

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.     

Effects of a commercial dose of L-tryptophan on plasma tryptophan concentrations and behaviour in horses

Reasons for performing the study: L‐tryptophan is a common ingredient in equine calmative products, but its effectiveness has not been demonstrated in horses. Hypothesis: To determine whether a commercial dose of L‐tryptophan increases plasma tryptophan and alters behaviour in horses fed a roughage or concentrate meal. Methods: L‐tryptophan (6.3 g) or placebo (water) was administered per os in a cross‐over design, to 12 Thoroughbred horses (503 ± 12.1 kg bwt), just before a meal of lucerne hay or oats. Plasma tryptophan was measured by gas chromatography. Horse behaviour was observed in an empty enclosure, then in the presence of an unfamiliar person and a novel object. Results: Total plasma tryptophan increased 3‐fold in both studies, peaking 1.5‐2 h after dosing. After the peak, tryptophan remained high for several hours if the horses had been fed hay, but fell sharply if fed oats, consistent with the glycaemic responses to these meals. However, the ratio of tryptophan to 4 large neutral amino acids (phenylalanine, tyrosine, leucine and isoleucine) increased in the tryptophan‐treated horses to a similar extent and for a similar duration, with both diets. The presence of a stranger or novel object increased heart rate (P<0.05), but caused no behavioural effects that were altered by tryptophan, regardless of the diet. Conclusions: Plasma tryptophan increases when tryptophan is administered at a dose used in some commercial products, but this is not reflected by marked behavioural changes in the horse. Potential relevance: Further work is required to refine behavioural tests and identify an effective dose of L‐tryptophan in the horse.     

Resting concentrations of the plasma free amino acids in horses following chronic submaximal exercise training

Six horses were conditioned on a treadmill at a constant speed of 5.6 km/hr on a 12.5% grade for gradually increasing periods of time over 14 days in order to determine the effect of repeated submaximal exercise on the concentrations of plasma free amino acids, protein metabolism, and plasma volume. Following 14-days of training, plasma volume increased (29%, P<0.05), as did total circulating content of plasma protein, albumin and urea. Urinary urea nitrogen excretion decreased (P<0.05) with exercise training. After the first week of training, the concentration of glycine had decreased (P<0.05) and the concentrations glutamic acid, arginine and alanine were increased (P<0.05) when compared to their corresponding pre-training (control week) levels. Compared to pretraining levels, there were decreases (P<0.05) in aspartic acid, histidine, arginine, valine, phenylalanine, isoleucine, and lysine, following the second week of training. Following a week of recovery, all resting concentrations of plasma free amino acids; when compared to their pretraining control; had decreased, with the exception of three nonessential amino acids (glutamic acid,serine, and glycine). Based upon the results of the present study, it would appear that exercise training produced a significant change in the amino acid and protein metabolism of the horse.