Effects of a nine-month endurance training programme on muscle composition in the horse

Muscle biopsy samples were collected from the middle gluteal muscle of seven horses undergoing a nine-month endurance training programme. Samples were collected before the programme began and again after three, six and nine months of training. A fifth sample was collected three months after training ceased. Serial muscle sections were reacted histochemically for myosin adenosine triphosphatase after either acid (pH 4.3 and 4.6) or alkaline (pH 10.3) pre-incubation, and muscle fibres identified as type I, IIA, IIB or IIC. The oxidative capacity of individual fibres was assessed, using the reduced nicotinamide dinucleotide tetrazolium reductase stain, and the number of intermyofibrillar capillaries adjacent to each fibre was counted after staining, using the alpha-amylase periodic acid Schiff technique. Biochemical analyses involved the fluorometric measurement of the enzymes citrate synthase, 3-hydroxy acyl CoA dehydrogenase and lactate dehydrogenase as markers of end terminal oxidative, beta oxidative and glycolytic potential, respectively. There was an increase in the percentage of type IIB fibres having high nicotinamide dinucleotide tetrazolium reductase staining after three months training. This increase persisted throughout the period of training and during the period without training. There was an increase in the number of capillaries adjacent to type IIB fibres after six and nine months training. These had returned to near pre-training numbers after three months without training. There were increases in the activities of citrate synthase and 3-hydroxy acyl CoA dehydrogenase after three months training. The activities of both enzymes continued to rise throughout training and the highest activities were attained after nine months.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of growth and training on muscle adaptation in Thoroughbred horses

OBJECTIVE: To determine the effect of growth and training on metabolic properties in muscle fibers of the gluteus medius muscle in adolescent Thoroughbred horses. ANIMALS: Twenty 2-year-old Thoroughbreds. PROCEDURE: Horses were randomly assigned to 2 groups. Horses in the training group were trained for 16 weeks, and control horses were kept on pasture without training. Samples were obtained by use of a needle-biopsy technique from the middle gluteus muscle of each horse before and after the training period. Composition and oxidative enzyme (succinic dehydrogenase [SDHI) activity of each fiber type were determined by use of quantitative histochemical staining procedures. Whole-muscle activity of SDH and glycolytic enzyme (phosphofructokinase) as well as myosin heavy-chain isoforms were analyzed biochemically and electrophoretically, respectively. RESULTS: The SDH activity of type-I and -IIA fibers increased during growth, whereas whole-muscle activity was unchanged. Percentage of type-IIX/B muscle fibers decreased during training, whereas that of myosin heavy-chain IIa increased. The SDH activity of each fiber type as well as whole-muscle SDH activity increased during training. An especially noticeable increase in SDH activity was found in type-IIX/B fibers. CONCLUSIONS AND CLINICAL RELEVANCE: Changes in muscle fibers of adolescent Thoroughbreds are caused by training and not by growth.The most noticeable change was for the SDH activity of type-IIX/B fibers. These changes in the gluteus medius muscle of adolescent Thoroughbreds were considered to be appropriate adaptations to running middle distances at high speeds.     

Effects of repeated biopsying on muscle tissue in horses

The collection of muscle samples in horses to estimate their performance potential, adaptations to training and diagnosis of exertional myopathies is of increasing interest. This study was designed to examine whether repeated biopsying of the gluteus medius muscle at 7 week intervals alters morphology and muscle fibre variables in the sampling area. The gluteus had been biopsied 3 times previously, each biopsy separated by a 7 week interval. Seven weeks after the last biopsy, this study was initiated. Percutaneous needle biopsies were taken from left and right muscles at 2 depths (2 and 6 cm) and at 2 sites for each depth (22 and 25 cm from the tuber coxae) of 7 mature Thoroughbreds. The 22 cm site was located close (2 cm caudal) to an area of the muscle that had been biopsied previously. The 25 cm site was 5 cm apart from this area. A total of 52 samples were available for histology, electron microscopy and immunohistochemistry. The latter was used to study percentages, sizes and capillaries of muscle fibre types. Most muscle samples collected (75%) had normal morphology, but the remaining 25% showed signs of muscle repair. In some circumstances, they showed regenerative signs of complete muscle repair, while in others ineffective muscle repair (scar formation) was evident. Nevertheless, sufficient normal tissue was always available for measuring routine muscle variables. Samples collected 3 cm laterally apart showed large differences with regard to muscle fibre type variables examined. These results show that repeated muscle samples in intervals of 7 weeks do not have effects that would impair evaluation of muscle fibre variables for diagnosis of effectivity of conditioning programmes. Furthermore, they emphasise that the only way of keeping variations through sample collection technique small is to standardise all biopsying techniques accurately.     

Effects of training on biochemical values in standardbred horses.

Effects of training at a regular, fixed, standard exercise load on venous lactic acid, mixed venous and arterial blood gases and pH, and serum muscle enzymes were determined on previously unconditioned, healthy, adult, Standardbred horses. Arterial and mixed venous blood gases, pH, and serum muscle enzymes did not change in a consistent manner during training. Venous lactic acid concentrations did increase significantly with training and may be of value for the biochemical evaluation of fitness in horses.     

Effect of show jumping training on the development of locomotory muscle in young horses

OBJECTIVE: To investigate whether training for show jumping that is commenced early after birth affects the characteristics of equine locomotory muscle. ANIMALS: 19 Dutch Warmblood horses. PROCEDURES: Horses were assigned to a trained or not trained (control) group. After weaning, training (free jumping [2 d/wk] that was alternated with a 20-minute period of exercise in a mechanical rotating walker [3 d/wk]) was started and continued until horses were 3 years old. Fiber type composition (determined from myosin heavy chain [MyHC] content), fiber area, diffusion index (area supplied by 1 capillary), citrate synthase activity, and Na(+),K(+)-ATPase content were assessed in gluteus medius muscle specimens collected at 0.5, 1, 2, and 3 years. RESULTS: Developmental changes included an increase in MyHC fiber type IIa and a decrease in type IIad; increases in fiber area, diffusion index, and citrate synthase activity; and a decrease in Na(+),K(+)-ATPase content. The MyHC fiber type I and type IId were detected in high and low proportions, respectively. Training increased Na(+),K(+)-ATPase content, but did not affect other variables. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, show jumping training at an early age resulted in increased Na(+),K(+)-ATPase content of the deep portions of the gluteus medius muscle. The lack of training effects on the other muscle characteristics can partly be explained by the fact that an appropriate (aerobic) fiber type composition was already established at training commencement. These data also suggested that the developmental changes in equine muscle represent sufficient adaptation to meet the demands of this specific training.     

Effects of stylopharyngeus muscle dysfunction on the nasopharynx in exercising horses

REASONS FOR PERFORMING STUDY: Nasopharyngeal collapse has been observed in horses as a potential cause of exercise intolerance and upper respiratory noise. No treatment is currently available and affected horses are often retired from performance. OBJECTIVE: To determine the effect of bilateral glossopharyngeal nerve block and stylopharyngeus muscle dysfunction on nasopharyngeal function and airway pressures in exercising horses. METHODS: Endoscopic examinations were performed on horses at rest and while running on a treadmill at speeds corresponding to HRmax50, HRmax75 and HRmax, with upper airway pressures measured with and without bilateral glossopharyngeal nerve block. RESULTS: Bilateral glossopharyngeal nerve block caused stylopharyngeus muscle dysfunction and dorsal nasopharyngeal collapse in all horses. Peak inspiratory upper airway pressure was significantly (P = 0.0069) more negative at all speeds and respiratory frequency was lower (P = 0.017) in horses with bilateral glossopharyngeal nerve block and stylopharyngeus muscle dysfunction compared to control values. CONCLUSIONS: Bilateral glossopharyngeal nerve anaesthesia produced stylopharyngeus muscle dysfunction, dorsal pharyngeal collapse and airway obstruction in all horses. POTENTIAL RELEVANCE: The stylopharyngeus muscle is probably an important nasopharyngeal dilating muscle in horses and dysfunction of this muscle may be implicated in clinical cases of dorsal nasopharyngeal collapse. Before this information can be clinically useful, further research on the possible aetiology of stylopharyngeus dysfunction and dysfunction of other muscles that dilate the dorsal and lateral walls of the nasopharynx in horses is needed.     

Effects of draught load exercise and training on calcium homeostasis in horses

This study was conducted to investigate the effects of draught load exercise on calcium (Ca) homeostasis in young horses. Five 2-year-old untrained Standardbred horses were studied in a 4-month training programme. All exercise workouts were performed on a treadmill at a 6% incline and with a constant draught load of 40 kg (0.44 kN). The training programme started with a standardized exercise test (SET 1; six incremental steps of 5 min duration each, first step 1.38 m/s, stepwise increase by 0.56 m/s). A training programme was then initiated which consisted of low-speed exercise sessions (LSE; constant velocity at 1.67 m/s for 60 min, 48 training sessions in total). After the 16th and 48th LSE sessions, SETs (SET 2: middle of training period, SET 3: finishing training period) were performed again under the identical test protocol of SET 1. Blood samples for blood lactate, plasma total Ca, blood ionized calcium (Ca(2+)), blood pH, plasma inorganic phosphorus (P(i)) and plasma intact parathyroid hormone (PTH) were collected before, during and after SETs, and before and after the first, 16th, 32nd and 48th LSE sessions. During SETs there was a decrease in ionized Ca(2+) and a rise in lactate, P(i) and intact PTH. The LSEs resulted in an increase in pH and P(i), whereas lactate, ionized Ca(2+), total Ca and intact PTH were not affected. No changes in Ca metabolism were detected in the course of training. Results of this study suggest that the type of exercise influences Ca homeostasis and intact PTH response, but that these effects are not influenced in the course of the training period.     

Effect of high intensity training on anaerobic capacity of middle gluteal muscle in Thoroughbred horses

We hypothesize that high intensity training for Thoroughbred horses that have been subjected to conventional training could further improve the metabolic properties of the middle gluteal muscle. Nine well-trained horses were subjected to high intensity (80-100% Vdot;O(2)max, 5 minx2) training for 12 weeks. Biopsy samples were obtained from the muscle before and after 4 and 12 weeks of training. Three of the 9 horses did not complete the training programme. In the remaining 6 horses, activities of succinic dehydrogenase (SDH), phosphofructokinase (PFK) and 3-hydroxy acyl CoA dehydrogenase (HAD), and the composition of myosin heavy chain isoforms were analyzed by biochemical techniques. After 12 weeks of training, a significant increase was found in PFK activity but not in the SDH and HAD activities. There were no significant changes in the composition of myosin heavy chain isoforms. The high intensity training in this study was effective at increasing glycolytic enzyme activity, indicating the possibility to improve anaerobic capacity, which potentially could contribute greatly to performance in Thoroughbred horses. This study also highlighted a fact that high intensity training should be given with the great care to prevent the skeletal muscle injuries.     

Variations in muscle fiber composition between successfully and unsuccessfully raced quarter horses

Muscle biopsy samples were removed from the m gluteus medius of 47 retired running Quarter Horse (QH) mares. Horses were separated based on bloodline, past racing history and percentage Thoroughbred (TB). The bred to run and raced (BRRA) and bred to run and not raced (BRNR) groups possessed a lower percentage (P<.01) of fast-twitch low oxidative (FT) fibers (BRRA 38.6 and BRNR 36.2±2.54 respectively). These horses also possessed a higher percentage (P<.01) of fast-twitch high oxidative (FTH) fibers (BRRA 52.5 and BRNR 48.5±2.54, respectively) than not bred to run and not raced (NBNR) horses. Horses that were bred to run had higher (P<.05) FTH to FT fiber ratios (1.44) than horses that were not bred to run (.94). Successful racehorses possessed a lower percentage (P<.05) of slow-twitch (ST) fibers and a higher percentage (P<.05) of FT fibers than unsuccessful racehorses. Success or failure was determined by Speed Index. No differences (P>.05) were found in the percentage FTH fibers between successful and unsuccessful horses.     

Effects of training on resting peripheral blood and BAL-derived leucocyte function in horses

In this study, the effects of prolonged, high intensity training on aspects of peripheral blood and bronchoalveolar lavage (BAL)-derived leucocyte function were evaluated in 8 horses. All horses undertook a 7 week endurance training programme, followed by 5 weeks of high intensity training (HIT). Thereafter, horses were divided into control (C) and overtraining (OT) groups. The frequency and intensity of training were increased more substantially for horses in the OT group. Training was terminated in week 32 when horses in the OT group demonstrated a significant performance reduction. Peripheral blood and BAL samples were collected from 4 horses in C and OT groups in training weeks 7, 11, 14, 18, 22, 28 and 32. Flow cytometric techniques were used to assess phagocytosis by peripheral blood neutrophils and pulmonary alveolar macrophages (PAM), and oxidative burst activity of neutrophils, PAM, peripheral blood and BAL-derived lymphocytes. Peripheral blood neutrophil phagocytosis (internalisation) increased during the initial HIT period and decreased from week 16 when the training workload was increased for both groups. The oxidative burst activity of peripheral blood neutrophils and lymphocytes similarly increased and then decreased in response to training. The oxidative burst activity of PAM was reduced towards the end of the overtraining phase of the programme. Pulmonary alveolar macrophage phagocytosis and oxidative burst activity of BAL-derived lymphocytes demonstrated no change throughout the course of the study. There was no difference in results obtained from C or OT group horses, suggesting that protracted HIT, rather than overtraining, was associated with impaired cell function. The detrimental effects observed in peripheral blood neitrophil and PAM function may indicate impaired nonspecific immunity which may adversely affect the health and performance of horses undergoing protracted periods of intense training.     

Effects of exogenous equine somatotropin on mineral balance in two-year-old horses in race training

This study examined the effects of exogenous equinesomatotropin (eST — Equigen) administration on mineral absorption and retention in two-year-old horses in race training. Sixteen Quarter Horse geldings were paired by age (ave.age=794 d), and one horse from each pair was assigned at random to either the eST treatment group or the control. The experiment was conducted over 112 days during which the horses were gentled to ride and trained on a dirt track in a regimen typical for race horses in training. At 28-day intervals, collections of total fecal and urinary output were made to determine effects on Ca, inorganic P, Mg, Cu and Zn mineral balance.Due to marginal and slightly deficient amounts (P<01)of Ca, P, Mg and Zn intake by days 84 and 112, a result of reduced feed intake to maintain a constant body condition in the presence of a decreased workload, mineral balance for the aforementioned minerals was only examined through day 56. In contrast to the previous minerals, Cu intake appeared adequate. Ca, P, Mg, Cu and Zn intake did not differ (P>.05) by treatment. The eST-treated horses increased (P<.05) Ca retention as a percent of Ca absorbed, had greater (P<.05) overall apparent efficiency of P absorption and greater (P<.05) apparent retention of P on day 56 than the control horses. The eST-treated horses had greater (P<.05) apparent Cu absorption on day 56, greater (P<.05) apparent Cu absorption efficiency on days 56 and 84, and there was a trend (P<.1) for the eST-treated horses to retain more Cu on day 56 than the control horses. Finally, the eST-treated horses retained a greater percent (P<.1) of absorbed Zn on day 56.     

The effect of physical training on skeletal muscle enzyme composition in pigs.

Running exercise in pigs results in an elevation of lactic acid in blood. This elevation in blood lactate does not occur in physically conditioned pigs. Activities of succinic dehydrogenase, fructose-l,6-diphosphate aldolase, lactate dehydrogenase and creatine Phosphokinase as well as the myoglobin content were determined in m. gastrocnemius from 6 ergometer-trained and 4 untrained pigs. The succinic dehydrogenase and myoglobin contents were significantly higher (P<0.01) in trained animals, whereas no changes were noted in the aldolase and creatine Phosphokinase contents. The lactate dehydrogenase showed somewhat reduced levels in the trained pigs. This was accompanied by an increased H/M subunit ratio. The results provide evidence for an increase in the maximal aerobic metabolism in trained pigs and that trained pigs to a higher extent can rely on an aerobic energy metabolism during running exercise.     

Effects of muscarinic receptor antagonists on acetylcholine-induced contractions of jejunal smooth muscle in horses

This study investigated the effects of a muscarinic type 1 (M(1)), 2 (M(2)), and 3 (M(3)) antagonists (4-DAMP, pirenzepine, and methoctramine, respectively) on acetylcholine (Ach)-induced contractions of longitudinal jejunal muscle strips of horses. Strips were irrigated with Krebs-Henseleit solution gassed with 95% O(2) and 5% CO(2), and the developed tension in response to Ach was recorded before and after incubation with increasing concentrations of 4-DAMP (10(-8)-10(-6) M), pirenzepine (10(-6)-10(-4) M), and methoctramine (10(-5)-10(-3) M). When competitive antagonism was characterized, the affinity constant (pA(2)) was calculated by Schild plots. A parallel rightward shift in the concentration-response curves was observed after 4-DAMP and pirenzepine. Methoctramine presented a dual effect on the concentration-response curves: lower concentrations induced a parallel rightward shift without altering the maximum intensity of contraction (E(max)), while the highest concentration increased slope of the concentration-response curve and increased E(max). The pA(2) for 4-DAMP and pirenzepine was 9.18 and 7.13, respectively. Acetylcholine-induced contractions of longitudinal jejunal smooth muscle are mediated mainly via M(3) receptors. The complex role of M(2) receptors in jejunal smooth muscle contractions was evident because methoctramine potentiated the contractile response to higher doses of Ach.     

Comparative study of muscle fiber type composition in the middle gluteal muscle of andalusian, thoroughbred and arabian horses

Samples were taken from the right middle gluteal muscle of 25 Andalusian horses, 20 Thoroughbreds and 10 Arabian horses, all aged between 5-and 12-years-old and not involved in training. Histochemical analysis was carried out using myosin ATPase after acid preincubation (pH 4.4) and NADH-tr in order to determine the percentage of different fiber types. Statistical analysis showed highly significant differences (P<0.001) in the proportion of type 1 and type 2 fibers between breeds. The Thoroughbred was found to have a significantly higher proportion of type 2A fibers (P<0.001) than the other 2 breeds, although differences in the proportion of type 2B fibers in different breeds were not found to be statistically significant. The Andalusian horse was found to have a higher proportion of fast-twitch fibers of low oxidative capacity (type FT) than the other 2 breeds (P<0.001). These results not only confirm those obtained in previous studies of different breeds of horses, but highlight the histochemical differences in the skeletal muscle of breeds of great commercial and sporting interest in Spain.     

Effects of two training protocols on Angiotensin I-converting enzyme (ACE) activity in horses

Reasons for performing study: Studies in man have shown a correlation between Angiotensin I‐converting enzyme (ACE) genetic polymorphisms, ACE activity in the blood and superior athletic performance in sports requiring endurance. It has been hypothesised that the same correlation occurs in horses. There is no information in the literature concerning the effects of training on ACE activity in equine plasma. Hypothesis: Exercise training influences the activity of circulating ACE and the response observed is dependent on the exercise protocol. Methods: Thirteen horses of mixed breeds were randomly allocated 2 different training protocols to be carried out for a period of 15 weeks. Blood samples were collected from each horse before the beginning of training to determine baseline values. Subsequent sampling took place every 15 days throughout the training phase and for 8 weeks of paddock rest. Angiotensin I‐converting enzyme activity was determined by automated spectrophotometry. Results: Training for 15 weeks significantly increased plasma ACE activity, irrespective of training protocol. Differences observed in ACE activity pattern between the 2 training protocols were not statistically significant. Increase in ACE activity peaked with maximum workload. As soon as training was interrupted, ACE levels significantly decreased. Conclusions and discussion: Exercise training affects levels of ACE activity in equine plasma. The mechanism for this is not yet elucidated, but cardiovascular adaptation to exercise and blood pressure changes might be involved in this regulation. Potential relevance: Exercise training produced a gradual increase in enzymatic activity and might warrant the use of ACE as a tool for fitness monitoring. Angiotensin I‐converting enzyme enzymatic activity in the plasma might be directly correlated to a change in genetic expression and that variability must be taken into account when evaluating results from horses undergoing a physical training programme.     

Effects of training on potassium homeostasis during exercise and skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged Dutch Warmblood horses

OBJECTIVE: To investigate the effects of moderate short-term training on K+ regulation in plasma and erythrocytes during exercise and on skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged horses. ANIMALS: Four 4- to 6-year-old and four 10- to 16-year-old Dutch Warmblood horses. PROCEDURE: The horses underwent a 6-minute exercise trial before and after 12 days of training. Skeletal muscle Na+,K(+)-ATPase concentration was analyzed in gluteus medius and semitendinosus muscle specimens before and after the 12-day training period. Blood samples were collected before and immediately after the trials and at 3, 5, 7, and 10 minutes after cessation of exercise for assessment of several hematologic variables and analysis of plasma and whole-blood K+ concentrations. RESULTS: After training, Na+,K(+)-ATPase concentration in the gluteus medius, but not semitendinosus, muscle of middle-aged horses increased (32%), compared with pretraining values; this did not affect the degree of hyperkalemia that developed during exercise. The development of hyperkalemia during exercise in young adult horses was blunted (albeit not significantly) without any change in the concentration of Na+,K(+)-ATPase in either of the muscles. After training, the erythrocyte K+ concentration increased (7% to 10%) significantly in both groups of horses but did not change during the exercise trials. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, the activation of skeletal muscle Na+,K(+)-ATPase during exercise is likely to decrease with age. Training appears to result in an increase in Na+,K(+)-ATPase activity in skeletal muscle with subsequent upregulation of Na+,K(+)-ATPase concentration if the existing Na+,K(+)-ATPase capacity cannot meet requirements.     

Effects of feeding meals with various soluble-carbohydrate content on muscle glycogen synthesis after exercise in horses

OBJECTIVES: To determine effects of feeding diets with various soluble-carbohydrate (CHO) content on rates of muscle glycogen synthesis after exercise in horses. ANIMALS: 7 fit horses. PROCEDURES: In a 3-way crossover study, horses received each of 3 isocaloric diets (a high soluble CHO [HC] diet, a low soluble CHO [LC] diet, or a mixed soluble CHO [MC] diet). For each diet, horses were subjected to glycogen-depleting exercise, followed by feeding of the HC, LC, or MC diet at 8-hour intervals for 72 hours. RESULTS: Feeding the HC diet resulted in a significantly higher glycemic response for 72 hours and significantly greater muscle glycogen concentration at 48 and 72 hours after exercise, compared with results after feeding the MC and LC diets. Muscle glycogen concentrations similar to baseline concentrations were detected in samples obtained 72 hours after exercise in horses when fed the HC diet. Rate of glycogen synthesis was significantly higher when horses were fed the HC diet, compared with values when horses were fed the MC and LC diets. Glycogen synthase activity was inversely related to glycogen content. Protein content of glucose transporter-4 was the lowest at 72 hours after exercise when horses were fed the HC diet. CONCLUSIONS AND CLINICAL RELEVANCE: Muscle glycogen synthesis was slower after glycogen-depleting exercise in horses, compared with synthesis in humans. Feeding HC meals after strenuous exercise hastened replenishment of muscle glycogen content, compared with results for feeding of LC and MC diets, by increasing availability of blood glucose to skeletal muscles.