Amino acid profile during exercise and training in Standardbreds

The objective of this study is to assess the influence of acute exercise, training and intensified training on the plasma amino acid profile.In a 32-week longitudinal study using 10 Standardbred horses, training was divided into four phases, including a phase of intensified training for five horses. At the end of each phase, a standardized exercise test, SET, was performed. Plasma amino acid concentrations before and after each SET were measured.Training significantly reduced mean plasma aspartic acid concentration, whereas exercise significantly increased the plasma concentrations of alanine, taurine, methionine, leucine, tyrosine and phenylalanine and reduced the plasma concentrations of glycine, ornithine, glutamine, citrulline and serine. Normally and intensified trained horses differed not significantly. It is concluded that amino acids should not be regarded as limiting training performance in Standardbreds except for aspartic acid which is the most likely candidate for supplementation.     

Plasma lactate and purine derivatives accumulation after exercise of increasing intensity in standardbred horses

The aim of the experiment was to study the relationship between plasma lactate and allantoin accumulation in horses undergoing five exercises differing in intensity and length. Twenty-five adult trotter horses were used (18 males, two castrated, and five females), housed in three training centers. The horses were assigned to five groups: slow trot, over 2000 m (Group 1); slow trot over 1600 m (Group 2); fast trot over 1600 m (Group 3); fast trot over 2000 m (Group 4); fast trot over 2400 m (Group 5). Plasma was obtained from blood sampled at rest, at the end of the bout of exercise and after 15 and 45 minutes from the end of the bout of exercise and analyzed for glucose, lactate, uric acid, free fatty acids (FFA) and allantoin concentrations. Accumulations of plasma lactate and allantoin (mmol/sec) were calculated as difference between end of exercise and rest and between 45 minutes sample and rest, respectively.Ranking the intensity of exercise using the lactate concentrations at the end of exercise, the level of exertion was highest for Group 3 horses and lowest for Group 5 horses (20.9 and 2.8 mmol/l, respectively). At the end of exercise, glucose concentrations were much higher for horses undertaking the more intensive exercise (Groups 3 and 4 compared to Group 2). FFA concentrations were highest at the end of exercise for Groups 2 and 3 and after 15 minutes for Groups 4 and 5. Plasma uric acid and allantoin concentrations peaked 15 and 45 minutes from the end of exercise, respectively, independently of exercise intensity. The relationship between accumulation of plasma allantoin (y, dependent variables) and lactate (x, independent variable) was non-linear: y=0.15−2.61^*x+68.3^*x² (r²=0.900; se=0.19). This suggests that allantoin accumulation could be used together with plasma lactate to calibrate the workload to muscle conditions to prevent muscle injury.     

High intensity exercise conditioning increases accumulated oxygen deficit of horses

High intensity exercise is associated with production of energy by both aerobic and anaerobic metabolism. Conditioning by repeated exercise increases the maximal rate of aerobic metabolism, aerobic capacity, of horses, but whether the maximal amount of energy provided by anaerobic metabolism, anaerobic capacity, can be increased by conditioning of horses is unknown. We, therefore, examined the effects of 10 weeks of regular (4-5 days/week) high intensity (92+/-3 % VO2max) exercise on accumulated oxygen deficit of 8 Standardbred horses that had been confined to box stalls for 12 weeks. Exercise conditioning resulted in increases of 17% in VO2max (P<0.001), 11% in the speed at which VO2max was achieved (P = 0.019) and 9% in the speed at 115% of VO2max (P = 0.003). During a high speed exercise test at 115% VO2max, sprint duration was 25% longer (P = 0.047), oxygen demand was 36% greater (P<0.001), oxygen consumption was 38% greater (P<0.001) and accumulated oxygen deficit was 27% higher (P = 0.040) than values before conditioning. VLa4 was 33% higher (P<0.05) after conditioning. There was no effect of conditioning on blood lactate concentration at the speed producing VO2max or at the end of the high speed exercise test. The rate of increase in muscle lactate concentration was greater (P = 0.006) in horses before conditioning. Muscle glycogen concentrations before exercise were 17% higher (P<0.05) after conditioning. Exercise resulted in nearly identical (P = 0.938) reductions in muscle glycogen concentrations before and after conditioning. There was no detectable effect of conditioning on muscle buffering capacity. These results are consistent with a conditioning-induced increase in both aerobic and anaerobic capacity of horses demonstrating that anaerobic capacity of horses can be increased by an appropriate conditioning programme that includes regular, high intensity exercise. Furthermore, increases in anaerobic capacity are not reflected in blood lactate concentrations measured during intense, exhaustive exercise or during recovery from such exercise.     

Effect of creatine supplementation on muscle metabolic response to a maximal treadmill exercise test in Standardbred horses

The aim of the present study was to investigate the effect of creatine (Cr) supplementation on muscle metabolic response in connection with a maximal treadmill exercise test, known to cause a marked anaerobic metabolic response and adenine nucleotide degradation. First, 6 Standardbred trotters performed a standardised maximal exercise test until fatigue (baseline test). The test used was an inclined incremental treadmill test in which the speed was increased by 1 m/s, starting at 7 m/s, every 60 s until the horse could no longer keep pace with the treadmill. After this baseline test, the horses were separated into 2 equal groups. One half received a dose of 25 g creatine monohydrate twice daily, and the other group were given the same dose of lactose (placebo). The supplementation period was 6.5 days, after which the maximal treadmill exercise test was performed again. A washout period of 14 days was allowed before treatments were switched between groups and a new supplementation period started. After this second supplementation period a new maximal exercise test was performed. After supplementation with creatine or placebo, horses were stopped after performing the same number of speed steps and duration of exercise as they had in the baseline test. Blood samples for analysis of plasma lactate, creatine (Cr), creatinine, hypoxanthine, xanthine and uric acid concentrations were collected at rest, during each speed step and during recovery. The total blood volume (TBV) was also determined. Muscle biopsies for analysis of muscle metabolites (adenosine triphosphate [ATP], adenosine diphosphate [ADP], adenosine monophosphate [AMP], inosine monophosphate [IMP], creatine phosphate [CP], lactate [La] and glycogen) were taken at rest, immediately post exercise and after 15 min recovery. The results showed no significant increase in plasma Cr or muscle total creatine concentration (TCr) after supplementation with Cr. At the end of exercise ATP and CP concentrations had decreased and IMP and lactate concentrations increased in muscle in all groups. Plasma lactate concentration increased during exercise and recovery and plasma uric acid concentration increased during recovery in all groups. No influence could be found in TBV after supplementation with creatine. These results show that creatine supplementation in the dosage used in this study had no influence on muscle metabolic response or TBV.     

Repeated post-exercise administration with a mixture of leucine and glucose alters the plasma amino acid profile in Standardbred trotters

Background

The branched chain amino acid leucine is a potent stimulator of insulin secretion. Used in combination with glucose it can increase the insulin response and the post exercise re-synthesis of glycogen in man. Decreased plasma amino acid concentrations have been reported after intravenous or per oral administration of leucine in man as well as after a single per oral dose in horses. In man, a negative correlation between the insulin response and the concentrations of isoleucine, valine and methionine have been shown but results from horses are lacking. This study aims to determine the effect of repeated per oral administration with a mixture of glucose and leucine on the free amino acid profile and the insulin response in horses after glycogen-depleting exercise.

Methods

In a crossover design, after a glycogen depleting exercise, twelve Standardbred trotters received either repeated oral boluses of glucose, 1 g/kg body weight (BW) at 0, 2 and 4 h with addition of leucine 0.1 g/kg BW at 0 and 4 h (GLU+LEU), or repeated boluses of water at 0, 2 and 4 h (CON). Blood samples for analysis of glucose, insulin and amino acid concentrations were collected prior to exercise and over a 6 h post-exercise period. A mixed model approach was used for the statistical analyses.

Results

Plasma leucine, isoleucine, valine, tyrosine and phenylalanine concentrations increased after exercise. Post-exercise serum glucose and plasma insulin response were significantly higher in the GLU+LEU treatment compared to the CON treatment. Plasma leucine concentrations increased after supplementation. During the post-exercise period isoleucine, valine and methionine concentrations decreased in both treatments but were significantly lower in the GLU+LEU treatment. There was no correlation between the insulin response and the response in plasma leucine, isoleucine, valine and methionine.

Conclusions

Repeated post-exercise administration with a mixture of leucine and glucose caused a marked insulin response and altered the plasma amino acid profile in horses in a similar manner as described in man. However, the decreases seen in plasma amino acids in horses seem to be related more to an effect of leucine and not to the insulin response as seen in man.     

Effect of prolonged branched-chain amino acid supplementation on metabolic response to anaerobic exercise in standardbreds

This study investigated the effect of prolonged BCAA supplementation on metabolic response to a 1600m run on treadmill in Standardbred trotters. Four trained Standardbreds were divided into two groups and assigned in a 2×2 Latin square design. Both groups were fed and exercised similarly: one group received an oral amino acids supplement (12 g leucine, 9 g isoleucine and 9 g valine) 30 minutes prior to exercise and immediately after, the other group received a placebo. The horses received the supplement 3 days per week for 5 weeks. In the last week horses performed an anaerobic exercise test on an inclined (3.5%) high-speed treadmill. The exercise consisted of a 15 minute warm-up phase immediately followed by a 1600 m run at maximal speed (heart rate > 200 beats/min). Blood samples were collected pre-exercise, after exercise and during recovery (10 min, 30 min, and 24 h), and analyzed for lactate, ammonia, total protein, urea, uric acid, creatinine, free fatty acids (FFA), creatine kinase (CK), lactate dehydrogenase (LDH), aspartate amino transferase (AST). Heart rate was continuously recorded during exercise and recovery. No statistical differences between the groups were observed for all the considered parameters. Nevertheless, BCAA supplementation resulted in a higher plasma ammonia and urea concentrations as reported in previous studies in humans and rats. These data suggest that a BCAA supplementation are not effective in enhancing performance in healthy and well-fed horses.     

Effect of exercise on the immune response of young and old horses.

OBJECTIVE: To compare exercise-induced immune modulation in young and older horses. ANIMALS: 6 young and 6 aged horses that were vaccinated against equine influenza virus. PROCEDURE: Venous blood samples were collected for immunologic assessment before and immediately after exercise at targeted heart rates and after exercise for determination of plasma lactate and cortisol concentrations. Mononuclear cells were assayed for lymphoproliferative responses and incubated with interleukin-2 (IL-2) to induce lymphokine-activated killer (LAK) cells. Antibodies to equine influenza virus were measured. RESULTS: Older horses had significantly lower proliferative responses to mitogens than younger horses prior to exercise. Exercise caused a significant decrease in lymphoproliferative response of younger horses, but not of older horses. Activity of LAK cells increased slightly with exercise intensity in younger horses. Cortisol concentrations increased in both groups after exercise; younger horses had higher concentrations after exercise at heart rates of 180 and 200 beats/min than those of older horses. Plasma lactate concentrations increased with exercise intensity but there were no differences between older and younger horses. Older horses had lower antibody titers to equine influenza virus than younger horses. Exercise did not affect antibody titers. CONCLUSION: Although lymphoproliferative responses and antibody titers of older horses were less than those of younger horses, older horses were more resistant to exercise-induced changes in immune function, possibly because of lower cortisol concentrations. CLINICAL RELEVANCE: Stress and aging are known to affect immune function. Older horses had reduced immune function, but were more resistant to exercise-induced immune suppression than younger horses.     

Effects induced by exercise on lymphocyte β-adrenergic receptors and plasma catecholamine levels in performance horses

The effect of dynamic exercise on complete blood cell count, lymphocyte β-adrenergic receptor and plasma catecholamine (adrenaline and noradrenaline) levels in horses performing different disciplines were investigated during rest and after exercise. Blood samples were collected from jumping horses (n=6), Arabian Endurance horses (n=6) and Standardbred trotters (n=6) before and immediately after competition. Dynamic exercise caused a significant increase in red blood cell count (Standardbred trotters: P=0.0012), haemoglobin concentration (jumping horses: P=0.001; Standardbred trotters: P=0.01), haematocrit percentage (Standardbred trotters: P=0.005), neutrophil percentage (jumping horses: P=0.0003), lymphocyte percentage (jumping horses: P=0.0003), monocyte percentage (Standardbred trotters: P=0.0008), lymphocyte β-AR numbers (jumping horses: P=0.01; Arabian Endurance horses: P=0.016; Standardbred trotters: P=0.05), plasma adrenaline concentration (Standardbred trotters: P=0.0001) and plasma noradrenaline levels (Standardbred trotters: P=0.003). It is concluded that acute increases in plasma catecholamine concentrations depended on the exercise performed and may induce up-regulation of β-AR in equine lymphocytes. However, the exact mechanism of β-AR up-regulation still remains unclear.     

Changes in intramuscular amino acid levels in submaximally exercised horses – a pilot study

The time‐dependent changes in intramuscular amino acid (AA) levels caused by exercise and by feeding a protein/AA supplement were analysed in nine horses. Horses were submitted to a total of four standardized exercise tests (SETs). Amino acid concentrations were determined prior to, immediately after, 4 and 18 h after exercise. The experiment was subdivided into two consecutive periods of 3 weeks. In each period two SETs were performed. In the second period, horses were given a protein/AA supplement within 1 h after exercise. Significant changes in mean plasma AA levels similar to previous studies were noted to be time‐dependent and to be associated with feeding the supplement. The intramuscular concentrations of the free AA in relation to pre‐exercise levels showed significant time‐dependent changes for alanine, asparagine, aspartate, citrulline, glutamine, glycine, isoleucine, leucine, methionine, serine, taurine, threonine, tyrosine and valine. Feeding the supplement significantly increased the 4 h post‐exercise intramuscular concentration of alanine, isoleucine, methionine and tyrosine. At 18 h after exercise, apart from isoleucine and methionine, levels were still increased and also those of asparagine, histidine and valine in relation to none treatment. Hence, it was concluded that AA mixtures administered orally to horses within 1 h after exercise increased intramuscular AA pool.     

The influence of betaine on untrained and trained horses exercising to fatigue

Because exercise fatigue has been associated with the accumulation of lactic acid, factors that influence lactate metabolism during exercise can potentially enhance performance. The objective of this study was to examine the effects of supplemental betaine on eight mature Thoroughbred horses before and after 8 wk of conditioning. The effects of betaine were tested in two cross-over design experiments, allowing each horse to receive both the control and betaine treatments at each fitness level. Ingestion of 80 mg of betaine/kg of BW for 14 d before exercise testing did not alter plasma lactate, glucose, free fatty acids (FFA), or triglyceride concentrations during exercise in the untrained or trained horses. A time x treatment interaction (P < .05) was observed for plasma lactate in untrained horses during recovery from exercise, and plasma lactate concentrations were lower (P < .05) at 60 min after exercise when untrained horses received betaine. Plasma FFA concentrations were lower (P < .05) before exercise and at 720 min after exercise when untrained horses received betaine. These data indicate that betaine may influence lactate metabolism following exercise in untrained horses; however, betaine does not seem beneficial for trained horses.     

Individual differences and repeatability of post-prandial changes of plasma-free amino acids in young horses

Few data are available on post-prandial changes of plasma amino acids (AAs) in horses and on the repeatability and the individual variance on different sampling days. The objective of the present study was to measure pre- and post-prandial concentrations of plasma AA in 10 yearling horses. Blood samples were taken on days 1 and 40 of the study before feeding of hay, oats and soya meal and over an 8 h post-prandial period in 2-h intervals. The plasma AAs were measured by high-pressure liquid chromatography after ortho-phthalaldehyde derivatization. Mean fasting concentrations of the AAs were not significantly influenced by the individuum and sampling day. Repeatability of the fasting AA levels in the individual horses on two different sampling days was only found for histidine, 3-methylhistidine, methionine, tryptophan and taurine. While the absolute post-prandial AA concentrations differed between sampling days, the relative changes were comparable. All AA concentrations except 3-methylhistidine increased after feeding by 13% to more than 200% of their fasting values if the combined data of both days were analysed. Four hours after feeding the concentrations of arginine, asparagine, lysine, leucine, isoleucine and threonine, decreased more than 20%. Histidine, methionine, phenylalanine, valine, tryptophan, glutamine, glycine, tyrosine and taurine concentrations decreased by less than 20%. Concentrations of aspartic acid, glutamic acid, ornithine, serine and citrulline remained elevated. Most AA approached the fasting concentrations at 8 h, only glycine increased between 6 and 8 h after meal and 3-methyl-histidine concentrations were constant throughout the entire period. In conclusion, the pre-prandial plasma AA in horses appeared less influenced by individuum or sampling day than post-prandial plasma AA concentrations. Therefore, plasma AA concentrations should be interpreted only under well-defined conditions, especially regarding the feeding regimen.     

补喂支链氨基酸对速步马1km速步赛成绩及赛前、赛后血浆生化指标的影响

本试验旨在通过研究补喂支链氨基酸对速步马1 km速步赛成绩及赛前、赛后血浆抗氧化指标以及激素、肌酸、葡萄糖、乳酸、肌酐含量的影响,为支链氨基酸在速步马训练、比赛中的应用提供参考数据。试验选取年龄相近(4岁左右)、体重相近[(457±50)kg]并经过严格训练的伊犁马公马8匹(速步赛用马),随机分为2组,分别为对照组、试验组,每组4匹。每天每匹马分别饲喂3 kg颗粒精料,苜蓿干草自由采食,在此基础上试验组每天每匹马补喂72 g支链氨基酸(由35.0 g亮氨酸、16.6 g异亮氨基酸、20.4 g缬氨酸组成),进行为期38 d(预试期7 d,正试期31 d)的补饲试验及训练试验。结果表明:补喂支链氨基酸可提高速步马的比赛成绩,同时显著提高速步马赛后30 min血浆总抗氧化能力及赛后24 h血浆超氧化物歧化酶活力(P0.05),但对血浆中激素、肌酸、葡萄糖、乳酸、肌酐含量无显著影响(P0.05)。由此得出,补喂支链氨基酸可缩短速步马比赛用时,提高速步马机体的抗氧化能力,但对血浆中激素、葡萄糖、肌酸、乳酸、肌酐含量无显著影响。     

Effects of muscle glycogen depletion on some metabolic and physiological responses to submaximal treadmill exercise.

The aim of this study was to investigate the effects of reduced muscle glycogen concentration on some physiological and metabolic responses during moderate intensity treadmill exercise in horses. Six Thoroughbred geldings were randomly allocated to 2 treatments (protocols A and B) or control in a 3 x 3 replicated Latin square design. In protocol A, horses performed low intensity exercise while horses in protocol B performed short bursts of high intensity exercise. Protocol A was designed to induce glycogen depletion mainly of slow twitch muscle fibers while protocol B aimed to deplete mainly fast twitch muscle fibers. Horses in the control group did not undergo exercise prior to the exercise test. Five hours after glycogen depletion, horses performed treadmill exercise at 60% VO2max at a treadmill slope of 10% until fatigue (20-30 min). The induced glycogen depletion prior to exercise had no significant effect on plasma glucose, insulin, or lactate concentrations during the exercise test, and there was no effect on glycogen utilization rate, although respiratory exchange ratios were lower in the glycogen-depleted groups. The VO2, heart rate and central blood temperature did not vary significantly between the protocols A and B and control throughout the exercise test. It was concluded that 20-30% depletion of glycogen concentration in the middle gluteal muscle resulted in a shift towards fat metabolism, but does not significantly affect heart rate, oxygen uptake, or concentrations of plasma glucose and lactate during moderate intensity exercise.     

Blood glutathione status and activity of glutathione‐metabolizing antioxidant enzymes in erythrocytes of young trotters in basic training

The aim of this study was to evaluate response of blood glutathione status and activity of glutathione‐metabolizing antioxidant enzymes in erythrocytes of young trotters in basic training. Nine untrained trotters (aged 16–20 months) were exposed to a 4‐month training program based on exercises at low‐to‐moderate intensity. The conditioning consisted of breaking the horses and running them on distances varying from 4 to 40 km a week. The workloads were increased on a 3‐week basis. Exercise intensity was monitored by measuring heart rate and blood lactate. Blood samples were collected at rest, before (RES0) and after (RESt) the conditioning period; moreover, on the latter occasion (on day 112 of training), the blood was also taken immediately after the routine exercise (EXE0) and 60 min thereafter (EXE60). The whole blood samples were analysed for the concentration of reduced, oxidized and total glutathione (GSH, GSSG and TGSH, respectively), while the activities of glutathione peroxidase (GPX) and glutathione‐disulfide reductase (GR) were determined in haemolysates. Additionally, the erythrocytic concentrations of oxidized nicotinamide adenine dinucleotide (NAD⁺) and its phosphate (NADP⁺) were measured. All investigated parameters except NAD⁺ and reduced/oxidized glutathione ratio (GSH/GSSG) changed during the training period. Following the effortm GPX, NADP⁺ and GSH/GSSG were significantly lower (p < 0.05, p < 0.01, p < 0.001, respectively) while GSSG was markedly higher than at rest (RESt). The drop in NADP⁺, low GSH/GSSG and high GSSG concentration were sustained at EXE60. Glutathione‐disulfide reductase activity was higher after the workout but only at EXE60 the increase in activity was significant. Despite the activities of the GSH‐GSSG cycle, enzymes were considerably higher after the training period, the elevated concentration of GSSG and significantly lower GSH/GSSG ratio in the post‐exercise measurements suggest that production of reactive oxygen species possibly exceeds the capacity of antioxidative defenses of immature trotters. A more balanced diet with additional antioxidant supplementation and a revision of the basic training protocol used herein are advised.     

The opioid haemorphin-7 in horses during low-speed and high-speed treadmill exercise to fatigue

The opioid neuropeptide haemorphin-7 was measured, by immunoreactivity, in Standardbred horses during low-speed (7 m/s) and high-speed (10 m/s) endurance exercises, lasting 49-58 and 12-16 min respectively. In parallel, heart rate, muscle temperature and plasma lactate concentrations were measured. The profile of the low-speed exercise showed significantly increased heart rate after 10 min [154 beats per minute (bpm)]. After the exercise, muscle temperature (42.1 degrees C) and plasma lactate (4.8 mmol/l) were significantly increased. The profile of the high-speed exercise was comparatively characterized by a higher increase of heart rate after 5 min (194 bpm) and higher increases of muscle temperature (43.2 degrees C) and lactate levels (15.8 mmol/l) after the exercise. The horses were probably exhausted by glycogen depletion in the low-speed exercise and by muscle pH decrease in the high-speed exercise. Haemorphin-7 increased significantly during the high-speed exercise (274.8 fmol/ml) but not during low speed (108.3 fmol/ml), coincident with the results of lactate. These results suggest that plasma haemorphin-7 is measurable in the horse by immunoreactivity, and that intense exercise stimulates release of this opioid. Such endogenous opioids are most likely involved in regulatory functions associated with pain, physical effort, inflammation, and blood pressure variation in horses, as have been established in other species.     

Muscle characteristics and plasma lactate and ammonia response after racing in Standardbred trotters: relation to performance

Blood samples from the jugular vein and muscle biopsies (gluteus medius) in 25 Standardbred trotters were obtained 5-10 min after racing. The biopsies were analysed for fibre type composition and enzymatic profile and blood samples for plasma lactate and ammonia concentrations. Muscle characteristics, plasma lactate and ammonia concentrations after racing were compared with each horse's individual performance index (IPI). The IPI is calculated annually from the individual horse's racing performance (% placing 1, 2 or 3, total annual earnings, average earning per start, and best racing record), respectively, converted to and expressed as a percentage deviation from the average record of the same sex and age group. The IPI values were 100-116. Plasma lactate concentrations were 15.0-42.7 mmol/l (mean 31.3 mmol/l) and ammonia concentrations 65-210 micromol/l (mean 141 micromol/l) after racing. Fibre type composition varied among horses (range 9-27% for Type I, 32-54% for Type IIA, and 27-46% for Type IIB). Fibre type composition, enzyme activities, plasma lactate and ammonia responses to racing were not correlated to IPI. Ten of the horses also performed a submaximal test on the track, consisting of 5 incremental 1000 m heats at approximate speeds of 9.1, 9.5, 10.0, 10.5, and 11.1 m/s. Immediately after each heat a blood sample was drawn from the jugular vein for plasma lactate analysis. Plasma lactate response to exercise differed between horses, but no correlation was seen with IPI. Muscle characteristics, plasma lactate and ammonia concentrations after racing and lactate response to a submaximal track test did not correlate with current race performance expressed as IPI in a group of horses with average or better performance capacity at the time of testing. Analysis of lactate and ammonia in blood after racing is not a valuable tool to predict an individual performance index.     

Biochemical and antioxidant changes in plasma and erythrocytes of pentathlon horses before and after exercise

Abstract: Physical exercise in the horse induces a series of normal physiological and biochemical adaptations. Increasing metabolism and oxygen uptake may induce oxidative stress in various organs. The aim of this study was to examine exercise-induced changes in some plasma and RBC biochemical and antioxidant variables in pentathlon horses. Blood samples were taken from 14 horses before, immediately after, and 24 hours after competing in two 1-minute runs of intense exercise over jumps. The peak intensity periods were preceded by a 20-minute warm-up and separated by a 20-minute break. The following plasma biochemical analytes were determined: total protein, uric acid, and lactate concentrations, and lactate dehydrogenase (LDH) and creatine kinase (CK) activities. Total antioxidant status (TAS) and the ferric reducing ability of plasma (FRAP) also were measured. Thiobarbituric acid-reactive substances (TBARS), reduced glutathione (GSH), and total protein concentrations, and glutathione peroxidase (GSHPx) and superoxide dismutase (SOD) activities were determined in RBC hemolysates. Significantly increased concentrations of total protein, lactate, and FRAP, and increased activities of CK and LDH were observed immediately postexercise compared with pre-exercise samples (P < .05). All results returned to approximately initial values after 24 hours of rest. RBC GSH and TBARS concentrations did not change immediately after exercise, but decreased after 24 hours of rest (P < .05). Plasma uric acid and FRAP values were positively correlated in a linear model ( r = .78). In summary, the type of exercise applied in this study, which can be considered quite usual for pentathlon horses, caused detectable biochemical and lipid peroxidative changes in plasma and RBCs. FRAP and TAS values changed in opposite directions, indicating that when antioxidant capacity is assessed using different methods, highly different results may be obtained.     

Hyperkalemic periodic paralysis, plasma lactate and exercise tolerance

The purpose of this, trial was to determine the effect of hyperkalemic periodic paralysis (HYPP) on exercise tolerance in Quarter Horses. Five HYPP affected and five nonaffected horses were matched for age, size, gender and reproductive status. HYPP status was diagnosed by DNA analysis and potassium chloride challenge testing. Plasma lactate concentration and heart rate were used as indicators of work intensity. Serum potassium concentrations were also monitored. Two exercise experiments were conducted, the first being forty-five minutes of slow, aerobic exercise (hacking) and the other being moderate, partially anaerobic exercise (galloping). Post-exercise the horses were cooled out by randomly assigning them to either forty minutes, of standing still or forty minutes of walking. Heart rates of HYPP affected and unaffected horses were not significantly different during exercise or recovery. Plasma lactate concentrations changed slightly following slow exercise and were significantly higher for HYPP affected horses (P=0.01).At the end of exercise, values were 1.4±0.2 mmol/L and 1.0±0.1 mmol/L for HYPP affected and unaffected horses, respectively. Following moderate exercise, plasma lactate concentrations were much greater, and the difference (P<0.001) between affected and unaffected horses was more marked: immediately following exercise concentrations were 10.6±1.8 and 6.2±1.0 mmol/L in affected and unaffected horses, respectively. The higher post-exercise plasma lactate concentrations in affected horses indicates increased anaerobic muscle metabolism. Serum potassium concentrations rose following exercise and significantly higher values were seen in horses that were walked rather than stood still post-exercise.Hyperkalemic periodic paralysis (HYPP) is a dominant autosomal genetic defect occurring in American Quarter Horses and related breeds. ^1–5 The condition is widely geographically distributed and has been estimated to affect 0.4% of all Quarter Horses.⁶ Signs include sporadic attacks of muscle fasciculation, muscle spasm, sweating and weakness. Oral administration of potassium chloride produces a more severe hyperkalemia in HYPP affected horses and induces clinical attacks.¹ Electromyography reveals widespread continuous, spontaneous, muscle contraction.² Some people believe that HYPP affected horses are suitable for riding⁷; others have expressed doubts about the safety of this practice and the exercise tolerance of affected horses. HYPP affected horses have an unstable muscle membrane potential causing random muscle fiber contractions,^2,8 which could antagonize purposeful movement. This, in combination with increases in extracellular potassium concentration occurring during exercise,^9–11 may cause affected horses to stumble or collapse while being ridden, posing a danger to both horse and rider. In HYPP affected people, potassium concentration rises approximately one to two hours after exercise during which time HYPP attacks can occur.^9,10 For these reasons, the exercise tolerance of HYPP affected horses and the effects of exercise on spontaneous HYPP attacks deserves investigation.We standardized the exercise test and chose heart rate and plasma lactate concentrations as indicatoors, of exercise tolerance and energy metabolism.¹² Heart rate is the major determinant of oxygen delivery to muscle and the rate rises with exercise intensity until it reaches a plateau at high velocities.^13,14 Several lines of investigation indicate that plasma lactate concentrations reflect muscle work. Lactate release from resting muscle is minimal but large amounts are released during strenuous exercise.^15–17 Lactate release increases with increasing muscle work.^16,18 During exercise, muscle oxygen consumption increases until eventually a plateau is reached after which no further increase in oxygen consumption in response to increased work loads is possible.¹⁸ At this point the mitochondrial electron chain transport system is operating at its maximal possible rate, energy production can only be supplemented by the use of inefficient anerobic glycolytic metabolism and the muscle starts to release lactate. If the concentrations of plasma lactate are high, the muscles are depending on anaerobic metabolism and are working closer to exhaustion^16,18–21The objectives of this experiment were twofold. One was to determine if HYPP adversely affects exercise performance. The second was to study the effects of cooling out, by either standing still or walking, on serum potassium and plasma lactate concentrations.     

Haemodynamic response to exercise in Standardbred trotters with red cell hypervolaemia

In order to evaluate the haemodynamic response to exercise in Standardbred trotters with red cell hypervolaemia (RCHV), 12 trotters with RCHV were compared with 9 normovolaemic (NV) trotters. Haemodynamic data were recorded during exercise at 4 different speeds on a treadmill. Oxygen uptake was determined with an open bias flow system. Pulmonary artery pressure (PAP), systemic artery pressure (SAP), heart rate, packed cell volume (PCV) and plasma lactate and haemoglobin ([Hb]) concentrations were measured. Arteriovenous O2 content difference, cardiac output, stroke volume, pulmonary vascular resistance (PVR) and total systemic resistance (TSR) were calculated. Oxygen uptake, arteriovenous O2 content difference, heart rate, cardiac output, stroke volume, TSR and lactate did not differ between groups. The RCHV horses had significantly higher both mean diastolic and systolic PAP compared to NV horses and this difference increased with higher workload. Further, a higher SAP, PVR, PCV and [Hb] were found in RCHV horses during the course of exercise. Eleven of the RCHV horses, but none of the NV, showed exercise-induced pulmonary haemorrhage on endoscopic examination. The increase in red cell volume, resulting in a high PCV and high total blood volume, is suggested to be an important contributor to both the increased blood pressures in pulmonary and systemic circulation during exercise and to the development of exercise-induced pulmonary haemorrhage in RCHV horses.     

A preliminary study on the changes in some potential markers of muscle-cell degradation in sub-maximally exercised horses supplemented with a protein and amino acid mixture

In this preliminary study, time-dependent changes in plasma CK and AST activity, tyrosine (Tyr), 3-methyl-histidine (3mHis), glucose and lactate concentrations were analysed in nine horses under two different conditions. Furthermore, intramuscular concentrations of Tyr, 3mHis and activities of cathepsin B, acid phosphatase (ACP), glucose-6-phosphate dehydrogenase (G6PDH) and mRNA expression of ubiquitin were determined at the same time. After studying the effects of exercise alone, the effects of exercise and feeding of an experimental protein/amino acid (AA) supplement were analysed. Horses were submitted to a total of four standardised exercise tests (SETs) of high intensity. Potential markers of muscle break down were determined prior to, immediately after, 4 and 18 h after exercise. The experiment was subdivided into two consecutive periods of 3 weeks. In each period, two SETs were performed. In the second period, horses were fed with the protein/AA supplement within 1 h after exercise. Significant changes in plasma, intramuscular Tyr levels and mRNA expression of ubiquitin were caused both by time in relation to exercise and by treatment with the protein/AA supplement. The experimental supplement significantly decreased the 4-h post-exercise expression of ubiquitin mRNA in muscle. Only a borderline increase of markers of lysosomal involvement was seen and CK and AST activity generally showed their normal post-exercise patterns. A clear post-exercise reduction of this CK activity, however, was not observed after supplementation with the protein/AA mixture. The current findings indicate that horses might benefit from protein and AA supplementation directly after training by decreasing post-exercise proteolysis. The results support that further studies should be performed to characterize changes in equine protein metabolism caused by exercise including underlying molecular mechanisms.