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)     

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.     

Effects of exercise on skeletal muscle and serum enzyme activities in pigs

In order to determine skeletal muscle and serum enzyme activities following exercise, six adult Landrace pigs were submitted to 10 min running on a treadmill (0.5 m/s, on a 12% gradient) and compared to six controls. Blood samples were obtained just before the exercise, immediately after, and 24 h, 48 h and 144 h after exercise. Muscle biopsies were taken from the longissimus dorsi and biceps femoris 24 h after exercise. Total lactic dehydrogenase (LDH) and LDH isoenzymes in muscle and serum were unchanged. In muscle homogenates, there was no difference in total creatine phosphokinase (CK) activity between the two groups. Total CK activity in the biceps femoris muscle represented only 59% of that observed in the longissimus dorsi muscle. A mean CK-MB value of 2.5% was found in the control group for both muscles but after exercise it was 9.5% (p<0.05) for the biceps femoris and 12.2% (p<0.01) of the longissimus dorsi muscle. In serum, the total CK (p<0.05), CK-MM (p<0.05) and CK-BB (p<0.05) increased immediately after the exercise, followed by a progressive decrease.     

Effects of post-exercise supplements on glycogen repletion in skeletal muscle

Post-exercise carbohydrate supplementation has been routinely used to enhance glycogen concentrations in skeletal muscle, particularly during multiple-day athletic events. Consumption of protein hydrolysates mixed with carbohydrate supplements during the post-exercise period may increase insulin response and cause glycogen repletion in skeletal muscle. A group of Alaskan sled dogs were selected to examine post-exercise supplementation in a paired crossover study design. The dogs were subjected to the same exercise regimen and provided one of three treatments-water, glucose polymers, or glucose polymers with protein hydrolysates-over a 2-month period. Parameters tested at various post-exercise time points included plasma insulin, glucagon and glucose concentrations, and skeletal muscle glycogen content to gain a better understanding of glucose metabolism and glycogen repletion. The results showed an enhanced insulin, glucose, and glucagon response immediately after supplementation and significantly increased glycogen concentrations in skeletal muscle within 24 hours when dogs received either of the glucose-containing supplements compared with water alone. There were no differences in the plasma parameters or skeletal muscle glycogen stores in dogs provided the glucose polymers alone or the glucose polymers plus protein hydrolysates. Thus, post-exercise carbohydrate supplementation increased muscle glycogen repletion, but inclusion of protein hydrolysates in the supplements provided no additional benefits.     

Fibre types,enzyme activities and substrate utilisation in skeletal muscles of horses competing in endurance rides

Venous blood samples and middle gluteal muscle biopsies were obtained from 21 horses taking part in 100 km or 50 km endurance rides. Group A consisted of seven horses competing over 100 km (four horses completed the ride), Group B consisted of the six horses that were among the 10 best over 50 km while the other eight horses of Group C completed 50 km at a slower speed. Blood lactate, glycerol and creatine kinase increased in all groups while aspartate aminotransferase levels were higher only in Group A. No change was found in blood glucose in Groups B and C while horses in Group A had lower levels after the ride. Neither fibre composition, fibre areas nor enzyme activities differed between the groups. Intramuscular glycogen content was similar before the ride in all groups. After the ride glycogen had decreased only half as much in Group C as compared to Groups A and B. Group C had a higher intramuscular triglyceride content at rest than Group B. The greatest decrease in triglyceride content after the ride was found in Group C. In Groups A and B many Type I, IIA and IIB fibres were glycogen depleted after the ride while in Group C mainly Type I and some Type IIA fibres were depleted. The results of this study show that intramuscular carbohydrate and lipid stores are both important fuels during endurance rides.     

Cardiovascular effects of submaximal aerobic training on a treadmill in Standardbred horses, using a standardized exercise test

Seven healthy, unexercised, previously trained, adult Standardbred horses were allotted to 2 groups and trained 78 days on a treadmill set at a 7 degree 30' angle. The groups were trained on different schedules, and the effects of training on heart rate, cardiac output, stroke volume, arteriovenous oxygen difference, systemic blood pressure, and venous lactic acid were determined. Measurements were made at rest, during exercise on the treadmill at rates of 55 m/min, 75 m/min, 100 m/min, and 154 m/min, and at 5 minutes after exercise (standardized exercise test). Heart rate and cardiac output decreased during the training period. Significantly slower heart rates were observed at 55 m/min by day 8, at 100 m/min and 154 m/min by day 36, at 1 minute after exercise by day 57, and at 5 minutes after exercise by day 78 (P less than 0.05). Stroke volume increased with exercise, but not significantly. The arteriovenous oxygen difference increased significantly (P less than 0.05) with each increase in work load. There was no significant increase with training, although an upward trend was recorded. Mean systemic blood pressure did not differ from resting with treadmill rates of 55 m/min, 75 m/min, or 100 m/min. It was greater at 154 m/min, although this was not significant. During exercise, the total peripheral resistance decreased to as little as 30% of its resting value. After exercise, diastolic and mean arterial blood pressures and peripheral resistance increased. Marked increases in blood volume and blood viscosity during exercise were closely related to the decrease in peripheral resistance. There was no significant effect of training on blood pressure. Venous lactic acid concentrations at rest were greater than those of the horses on the treadmill at rates of 55 m/min, 75 m/min, and 100 m/min and at 5 minutes after exercise on days 1, 8, and 15. Subsequently, they were not different from resting values. Differences in the effects of the different training programs could not be detected.     

Histochemical properties of muscle fibres types and enzyme activities in skeletal muscles of Standardbred trotters of different ages

Fibre characteristics and enzyme activities were determined for the gluteus, semitendinosus, vastus lateralis and triceps brachii muscles of 55 Standardbred trotters of different ages. Four fibre types (I, IIA, IIB, IIC) were demonstrated by histochemical staining of myofibrillar adenosine triphosphatase after preincubation at different pH values. Type II fibres predominated in all the muscles and the type IIA/IIB ratio was higher in horses over 5 years than in younger horses, except in the vastus in which the IIA/IIB ratio did not change with age. The vastus had the highest proportion of type IIA fibres and the semitendinosus the highest proportion of type IIB fibres. Histochemical demonstration of NADH dehydrogenase disclosed that almost 100 per cent of the type IIA and many of the type I and IIB fibres were medium-stained; the remaining type I fibres were darkly stained and the type IIB fibres lightly stained. In older horses more fibres were stained for NADH dehydrogenase. The activity of triosephosphate dehydrogenase decreased that that of 3-hydroxy-acyl-coA dehydrogenase and citrate synthase increased in all the muscles except the vastus with increasing age. The greatest increase in oxidative capacity occurred in the gluteus and triceps. Training, rather than age, was regarded as the factor inducing these changes. The results emphasise that histochemical data are only semiquantitative, and there are apparent discrepancies in the intensities of histochemical staining and the biochemical evaluation of various enzymes.     

Responses to submaximal treadmill exercise and training in the horse: changes in haematology, arterial blood gas and acid base measurements, plasma biochemical values and heart rate

Four standardbred horses with subcutaneously relocated carotid arteries were given a seven week training programme of treadmill exercise at a gradient of 19 per cent in order to assess if there were any effects of exercise and training on haematology, arterial blood gas and acid base measurements, plasma biochemistry and heart rate. The exercise consisted of one minute walking at 110 metres/minute followed by five minutes trotting at 200 metres/minute, twice daily in the first week. The period of trotting exercise was increased by one minute per week so that by the seventh week the horses were being given 12 minutes trotting twice daily. Before training commenced venous blood samples, for complete blood counts and plasma biochemistry, and arterial samples, for blood gas, acid base and lactate measurements, were taken at rest, after five minutes and 15 minutes of treadmill exercise (200 metres/minute) and 30 minutes and 60 minutes after completing the exercise. Heart rate was measured by telemetric electrocardiogram at similar intervals. This exercise test and blood collection were repeated after one, three, five and seven weeks of training. The only significant changes were a decrease in exercise lactate with training, increases in exercise and recovery total protein. The haematological response to treadmill exercise included an increase in certain red cell parametes and a leucocytosis which was caused by both a neutrophilia and a lymphocytosis. These effects had largely disappeared by 30 minutes after exercise and all values had returned to resting values by one hour after exercise.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

The influence of the PRKAG3 mutation on glycogen,enzyme activities and fibre types in different skeletal muscles of exercise trained pigs

Background

AMP-activated protein kinase (AMPK) plays an important role in the regulation of glucose and lipid metabolism in skeletal muscle. Many pigs of Hampshire origin have a naturally occurring dominant mutation in the AMPK γ3 subunit. Pigs carrying this PRKAG3 (R225Q) mutation have, compared to non-carriers, higher muscle glycogen levels and increased oxidative capacity in m. longissimus dorsi, containing mainly type II glycolytic fibres. These metabolic changes resemble those seen when muscles adapt to an increased physical activity level. The aim was to stimulate AMPK by exercise training and study the influence of the PRKAG3 mutation on metabolic and fibre characteristics not only in m. longissimus dorsi, but also in other muscles with different functions.

Methods

Eight pigs, with the PRKAG3 mutation, and eight pigs without the mutation were exercise trained on a treadmill. One week after the training period muscle samples were obtained after euthanisation from m. biceps femoris, m. longissimus dorsi, m. masseter and m. semitendinosus. Glycogen content was analysed in all these muscles. Enzyme activities were analysed on m. biceps femoris, m. longissimus dorsi, and m. semitendinosus to evaluate the capacity for phosphorylation of glucose and the oxidative and glycolytic capacity. Fibre types were identified with the myosin ATPase method and in m. biceps femoris and m. longissimus dorsi, immunohistochemical methods were also used.

Results

The carriers of the PRKAG3 mutation had compared to the non-carriers higher muscle glycogen content, increased capacity for phosphorylation of glucose, increased oxidative and decreased glycolytic capacity in m. longissimus dorsi and increased phosphorylase activity in m. biceps femoris and m. longissimus dorsi. No differences between genotypes were seen when fibre type composition was evaluated with the myosin ATPase method. Immunohistochemical methods showed that the carriers compared to the non-carriers had a higher percentage of type II fibres stained with the antibody identifying type IIA and IIX fibres in m. longissimus dorsi and a lower percentage of type IIB fibres in both m. biceps femoris and m. longissimus dorsi. In these muscles the relative area of type IIB fibres was lower in carriers than in non-carriers.

Conclusions

In exercise-trained pigs, the PRKAG3 mutation influences muscle characteristics and promotes an oxidative phenotype to a varying degree among muscles with different functions.     

Influence of the three RN genotypes on chemical composition, enzyme activities, and myofiber characteristics of porcine skeletal muscle.

An experiment was conducted to evaluate the effects of the RN genotype on skeletal muscle characteristics in pigs sharing otherwise the same polygenic background. Animals were genotyped for RN on the basis of RTN (Rendement Technologique Napole) records using segregation analysis methods. Samples of longissimus (L) and semispinalis capitis (S) muscles were taken from 39 rn+/rn+, 38 RN-/rn+ and 37 RN-/RN- pigs slaughtered at 108 +/- 8.6 kg live weight. Activities of lactate dehydrogenase (LDH), citrate synthase (CS), and beta-hydroxy-acyl-coenzyme A dehydrogenase (HAD) were measured on both muscles to assess glycolytic, oxidative, and lipid beta-oxidation capacities, respectively. Histological examinations and chemical analyses were performed on L muscle. The energetic metabolism of the white L muscle was more oxidative in RN-/RN- than in rn+/rn+ pigs, as shown by increased CS and HAD activities (P < .001), decreased LDH activity (P < .001), larger cross-sectional area of IIA (P < .05) and IIB-red (P < .05) fibers, higher relative area of IIA fibers ( P < .05), and lower relative area of IIB-white fibers (P < .001). No significant difference was found between heterozygous and homozygous carriers of the RN- allele, except for CS activity, which was lower in RN-/rn+ than in RN-/RN- pigs. In L muscle, the RN- allele led to a large increase in glycolytic potential (+3.5 phenotypic SD between homozygotes) and lightness (+.7 SD), and a decrease in ultimate pH, dry matter, and protein contents (-1.7 to -2 phenotypic SD for these three traits), with an almost completely dominant effect. No differences were found between genotypes for intramuscular fat and hydroxyproline contents. In the red S muscle, the presence of RN- had no influence on enzyme activities. These results indicate that the RN genotype greatly influences compositional and histochemical traits and metabolic enzyme activities in a muscle type-dependent manner, with a completely or incompletely dominant effect of the RN- allele.     

Effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle of male broilers

1. The effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle were evaluated using male broilers of Red Cornish x New Hampshire stock, reared on diets of high nutritional value for up to 80 d (H80d) and low nutritional value for up to 80 d (L80d, same age as H80d) or 95 d (L95d, same body weight as H80d). 2. The total live weight and the weight of pectoralis muscle were lower in L80d than in both H80d and L95d. The muscle weight as a percentage of live weight was 8.7% in L80d, 10.7% in H80d and 11.5% in L95d. 3. Pectoralis muscle was composed only of type IIB myofibres and showed no differences in myofibre type composition among the chicken groups. The largest diameter of type IIB myofibres was observed in L95d, followed by H80d and the smallest in L80d. 4. The total amount of intramuscular collagen did not differ among the chicken groups (1.92 to 1.99 mg/g). Types I and III collagens were immunohistochemically detected in both the perimysia and endomysia. The thin perimysia around the primary myofibre fascicles showed larger width in H80d than L80d and L95d, and also the thick perimysia around the secondary fascicles in H80d than L80d. 5. The collagen structure of the perimysium was most developed in H80d, followed by L95d and on the least in L80d. The development of perimysial collagen fibres could be enhanced by a rapid growth rate of the muscle induced by high nutritional level and depressed by a slow growth rate with low nutritional foods. 6. The endomysial collagen architecture was observed as a felt-like tissue of the fibril bundles with many slits. The thinnest endomysial wall was observed in L80d, followed by H80d and the thickest in L95d. 7. From these results, it was indicated that foods of high nutritional value could enhance growth of the pectoralis muscle of broilers, and this is accompanied by hypertrophy of the type IIB myofibres and development of the perimysial collagen architecture.     

Effects of submaximal exercise on adenine nucleotide concentrations in skeletal muscle fibers of horses with polysaccharide storage myopathy

OBJECTIVE: To determine whether disruption of adenine triphosphate (ATP) regeneration and subsequent adenine nucleotide degradation are potential mechanisms for rhabdomyolysis in horses with polysaccharide storage myopathy (PSSM) performing submaximal exercise. ANIMALS: 7 horses with PSSM and 4 control horses. PROCEDURES: Horses with PSSM performed 2-minute intervals of a walk and trot exercise on a treadmill until muscle cramping developed. Control horses exercised similarly for 20 minutes. Serum creatine kinase (CK) activity was measured 4 hours after exercise. Citrate synthase (CS), 3-OH-acylCoA dehydrogenase, and lactate dehydrogenase activities prior to exercise and glucose-6-phosphate (G-6-P) and lactate concentrations before and after exercise were measured in gluteal muscle specimens. Adenine triphosphate, diphosphate (ADP), monophosphate (AMP), and inosine monophosphate (IMP) concentrations were measured before and after exercise in whole muscle, single muscle fibers, and pooled single muscle fibers. RESULTS: Serum CK activity ranged from 255 to 22,265 U/L in horses with PSSM and 133 to 278 U/L in control horses. Muscle CS activity was lower in horses with PSSM, compared with control horses. Muscle G-6-P lactate, ATP, ADP, and AMP concentrations in whole muscle did not change with exercise in any horses. Concentration of IMP increased with exercise in whole muscle, pooled muscle fibers, and single muscle fibers in horses with PSSM. Large variations in ATP and IMP concentrations were observed within single muscle fibers. CONCLUSIONS AND CLINICAL RELEVANCE: Increased IMP concentration without depletion of ATP in individual muscle fibers of horses with PSSM during submaximal exercise indicates an energy imbalance that may contribute to the development of exercise intolerance and rhabdomyolysis.     

The effect of skeletal muscle needle biopsy on blood constituents, muscle glycogen and heart rate of cattle

Muscle samples from Friesian bulls were obtained using the Bergstr?m percutaneous biopsy needle. The samples (100 to 200 mg) were suitable for metabolite and enzymic analyses and histological examination in relation to meat quality. The physiological response to biopsy was assessed by measuring serum free fatty acid and blood glucose concentrations, plasma creatine kinase activity and heart rate. Muscle glycogen concentration was also measured. These parameters were unchanged by the experimental procedure, with the exception of blood glucose and heart rate which showed significant increases. The experimental animals showed no adverse effects after repeated biopsy and it is concluded that the technique is suitable for investigating beef muscle metabolism in vivo, and does not cause an unacceptable degree of stress.     

Effects of age and castration on activities of calpains and calpastatin in sheep skeletal muscle.

The objectives of this experiment were to assess effects of animal age and castration on activities of calpain I, calpain II, and calpastatin in sheep skeletal muscle. Ten newborn male lambs (2.9 kg), six weaned wethers (23.2 kg), six weaned rams (22.2 kg), six market wethers (55.4 kg), and six market rams (60.2 kg) were slaughtered and samples of biceps femoris were taken for assay of calpain I (micromolar calcium-dependent proteinase), calpain II (millimolar calcium-dependent proteinase), and calpastatin. Preweaning weight gain was similar for rams and wethers; however, postweaning ram growth exceeded (P less than .05) that of wethers. Ram biceps femoris weights at market were greater than those of wethers (P less than .05). Irrespective of age or gender, activity of calpain II was two- to threefold greater than that of calpain I. Muscle calpastatin activity was severa fold higher than calpain I and II activities. Activities of calpains and calpastatin declined (P less than .05) between birth and weaning. A portion of these losses were due to a dilution effect caused by accumulation of muscle proteins. Neonatal attenuation of calpain activities may underlie age-related attenuation of fractional rates of muscle protein degradation. Although ram muscle growth exceeded that of wethers, no differences (P greater than .05) in activities of muscle calpains or calpastatin were detected between these two groups at weaning or at market weight. Hence, castration did not influence lamb muscle growth by altering muscle calpain or calpastatin activities.     

Effects of training on Na, K-ATPase contents in skeletal muscle and K homeostasis of African draught bulls and cows

In semiarid parts of Africa animal traction is still one of the most reliable sources for rural work power. However, draught animals have to produce most of their work power at an unfavourable moment of the year that is at the end of the dry season when feedlot is scare. To improve their condition prior to the planting season, a short training could help. The effect of training can be expressed by the changes in contents of Na(+), K(+)-pumps in the muscle cell membrane. After a training period of 15 days all cattle showed a mean increase in Na(+), K(+)-ATPase of 24% (P < 0.01) in the semitendinosus muscle of the hind leg, whereas the control group showed no change. Bulls demonstrated already after 8 days of training an increase of 20% (P < 0.05). The principal factor responsible for this up-regulation of the Na(+), K(+)-pumps is most probably the excitation of muscles during exercise. In the course of the 15 days training period, the surge of plasma K(+) in during exercise showed a tendency to decrease, but this was not significant. Nevertheless, the reduced elevations of plasma [K(+)] may delay the moment of fatigue and so improve endurance. In conclusion, a training period of 8-15 days improves the contents of Na(+), K(+)-pumps and so the possible work output of draught cattle.