Effects of exercise and glucose administration on content of insulin-sensitive glucose transporter in equine skeletal muscle

OBJECTIVES: To characterize insulin-sensitive glucose-transporter (GLUT-4) protein in equine tissues and determine effects of exercise and glucose administration on content of GLUT-4 protein in equine skeletal muscle. SAMPLE POPULATION: Tissue samples from 9 horses. PROCEDURES: Western blot analyses were performed on crude membrane preparations of equine tissues to characterize GLUT-4. In a crossover, randomized study, horses were strenuously exercised for 3 consecutive days and then administered 13.5% glucose or isotonic saline (0.9% NaCl; control) solution, i.v., at similar infusion rates for 12.1 hours. Samples were collected from the middle gluteal muscle before and after exercise and 10.1 hours after completion of an infusion and used for measurements of glycogen concentration and total content of GLUT-4 protein. RESULTS: Immunoblot analyses detected specifically immunoreactive bands for GLUT-4 in insulin-sensitive tissues. Content of GLUT-4 protein in skeletal muscle increased significantly by 27.3 and 12.3% 22.2 hours after exercise for control and glucose groups, respectively. Intravenous infusion of glucose resulted in a significantly higher rate of glycogenesis, compared with results for the control group (mean +/- SD, 3.98 +/- 0.61 and 1.47 +/- 0.20 mmol/kg/h, respectively). Despite enhanced glycogenesis, we did not detect an increase in content of GLUT-4 protein after glucose infusion, compared with values after exercise. CONCLUSIONS AND CLINICAL RELEVANCE: GLUT-4 protein was expressed in equine skeletal and cardiac muscles. Exercise increased total content of GLUT-4 protein in skeletal muscle, and replenishment of muscle glycogen stores after glucose infusion attenuated the exercise-induced increase in the content of GLUT-4 protein in equine skeletal muscle.     

Effects of cooldown methods and durations on equine physiological traits following high-intensity exercise

This study was initiated to investigate the physiologic effects of cooldown methods and durations on recovery following high-intensity exercise (3000 m, 10 m/s) of 25 Jeju crossbred horses. Heart rate (HR) was measured and blood samples were collected for glucose, blood lactate concentration, packed cell volume (PCV), total protein (TP), and hemoglobin (Hb) analysis. The cooldown methods employed involved walk rest (WR) or trot rest (TR), and the durations compared were 15, 30, and 60 min. A passive rest group (PR) was used as a control. According to the analysis, HR decreased after a 15 min rest in all groups, showing a faster recovery in the active rest groups, WR and TR, than in the PR group after 30 min (P < 0.05). In the case of glucose, the decrease was faster in the WR15 group then the WR30 group suggesting that active rest is more effective in controlling this parameter (P < 0.05). As for lactate, TR15 showed a 75% decrease which was a significantly positive effect. There were no differences in PCV between the groups. Horses in the PR group showed faster recovery for TP and Hb than those in the active rest groups (P < 0.05), while lactate removal was faster in active rest groups than in the PR group, suggesting that the cooldown process plays an important role in recovery. This study showed that the cooldown method and duration employed after high-intensity exercise of horses makes a difference with regard to their physiologic status. Also, 15 min rest after exercise appears to be the most significant period in terms of duration. In conclusion, active rest was necessary for rapid recovery and a 15 to 30 min cooldown walk or trot was beneficial to the horses in which physical fatigue had accumulated. In particular, the active exercise method of trotting was found to be more effective than walking to remove lactate. In addition, the cooldown duration is expected to be more effective when adjusted according to the individual training state of the horse.     

Effects of beta-aminopropionitrile fumarate and exercise on equine tendon healing: Gross and histological aspects

In the present study, the effects of intralesional injections of beta-aminopropionitrile fumarate (BAPN-F) was assessed in equine experimental tendinitis. BAPN-F is a lathyrogen which inhibits, crosslinking of collagen, permitting more time for parallel reorientation of the repair tissue. Sixteen two-year-old Arabian horses without health problems were used in this experiment. The animals were divided into two groups: group one was left in box rest, and group two was submitted to controlled exercise during the experiment. Tendinitis was induced with collagenase in the superficial flexor tendon of both forelimbs. Twenty days after the induction of tendinitis, intralesional treatment with BAPN-F was performed and the contralateral limbs received saline. A biopsy was obtained and gross and histopathological analysis was performed on the 150th day of the experiment. The collagen fibrillar alignment pattern in the healing area was better in the BAPN-F group submitted to controlled exercise than in the other group, as observed by sonographic and histopathologic examination. The present results indicate that BAPN-F in combination with controlled loading improved scar remodeling and tendon wound collagen maturation.     

Effects of dietary yeast culture supplementation during the conditioning period on equine exercise physiology

Two groups of previously unconditioned young adult horses participated in 6 weeks of gradually increasing exercise on an inclined plane treadmill while receiving a cornoats-hay diet with or without a commercially available dietary yeast culture preparation. Forced treadmill exercise at a workload of 11.98 j/kg/m, equivalent to a workrate of 18.34 j/sec/kg and an estimated ground speed of 5.36 m/sec, began at 5 minutes per day (2.75 Mjoules/500 kg body-weight) and was increased by 5 minutes per week to a maximum of 35 minutes per day (19.25 Mjoules/500 kg) after 6 weeks. Treadmill exercise increased venous plasma lactate concentrations in direct proportion to the duration of an exercise bout, but the increases tended to be smaller after a given amount of work as the horses became conditioned. At the end of 35 minutes of exercise, plasma lactate concentrations averaged 30.08 mg/dl in the supplemented horses and 41.29 mg/dl in the unsupplemented horses (p<.01). Plasma glucose concentrations decreased significantly and triglyceride concentrations increased significantly in both groups as exercise duration exceed 10 minutes. Changes in plasma glucose concentrations were not significantly affected by yeast culture supplementation, while the supplemented horses exhibited somewhat slower rates of increased plasma triglyceride concentrations. During the 35-minute exercise bouts, significantly lower heart rates were recorded in the supplemented horses during the first 5 and the final 10 minutes of the workouts (p<.01), suggesting an enhanced state of athletic fitness. The digestible energy required for work (Mcal/500 kg bodyweight) was calculated to be 0.454 (Mcal/Mjoule) (Mjoules of work/500 kg bodyweight) + 0.024 Mcal/500 kg bodyweight (r²=0.95), with an efficiency of converting dietary DE to work of 53% for both groups of horses. Although the exercise challenges to these horses were not severe, these results suggest that dietary yeast culture supplementation of horses entering into conditioning programs may well enhance athletic training.     

Effects of exercise on tenocyte cellularity and tenocyte nuclear morphology in immature and mature equine digital tendons

REASON FOR PERFORMING STUDY: The injury-prone, energy-storing equine superficial digital flexor tendon (SDFT) of the mature performance horse has a limited ability to respond to exercise in contrast with the noninjury-prone, anatomically opposing common digital extensor tendon (CDET). Previous studies have indicated low levels of cellular activity in the mature SDFT, but in foal tendons the tenocytes may still have the ability to adapt positively to increased exercise. OBJECTIVES: To measure tenocyte densities and types in histological sections from the SDFT and CDET of horses from controlled long-term, short-term and foal exercise studies. METHODS: Specimens were collected from mid-metacarpal segments of the CDET and SDFT for each horse and processed for histology; central and peripheral regions of the SDFT cross-section were analysed separately (SDFTc, SDFTp). Tenocyte nuclei were counted in a total area of 1.59 mm(2) for each tendon region in each horse. Each nucleus was classified as type 1 (elongate and thin), type 2 (ovoid and plump) or type 3 (chondrocyte-like); type 1 cells are proposed to be less synthetically active than type 2 cells. RESULTS: No significant differences were noted between exercise and control groups in any of the studies, with the exception of an exercise-related reduction in the proportion of type 1 tenocytes for all tendons combined in the long-term study. There were tendon- and site-specific differences in tenocyte densities and proportions of type 1 and 2 cells in all 3 studies. CONCLUSIONS AND POTENTIAL RELEVANCE: There was no indication that exercise increased tenocyte density or proportions of the (theoretically) more active type 2 cells in immature horses (short-term and foal studies), perhaps because the training regimens did not achieve certain threshold strain levels. In the foal study these findings can still be interpreted positively as evidence that the training regimen did not induce subclinical damage.     

Methemoglobinemia and eccentrocytosis in equine erythrocyte flavin adenine dinucleotide deficiency

This report describes erythrocyte biochemical findings in an adult Spanish mustang mare that exhibited persistent methemoglobinemia, eccentrocytosis, and pyknocytosis that were not related to the consumption or administration of an exogenous oxidant. The methemoglobinemia was attributed to a deficiency in cytochrome-b5 reductase (Cb5R) activity, and the eccentrocytes and pyknocytes were attributed to a marked deficiency in reduced nicotinamide adenine dinucleotide phosphate-dependent glutathione reductase (GR) activity that resulted in decreased reduced glutathione concentration within erythrocytes. The GR activity increased to a near-normal value after addition of flavin adenine dinucleotide (FAD) to the enzyme assay, indicating a deficiency of FAD in erythrocytes. The methemoglobinemia, eccentrocytosis, and pyknocytosis were attributed to deficiency of FAD in erythrocytes because the GR and Cb5R enzymes use FAD as a cofactor. This deficiency in FAD results from a defect in erythrocyte riboflavin metabolism, which has not been documented previously in animals.     

Effect of exercise on the partitioning of equine respiratory resistance

Pleural, tracheal, pharyngeal and mask pressures as well as airflow and tidal volume of five ponies on a treadmill (incline 8.3 degrees) were recorded simultaneously while resting, walking, trotting slowly, trotting fast, after standing for 30 secs and 5 mins after the end of the exercise. The curves obtained were used to calculate total pulmonary resistance (RL), lower airway resistance (RLA) and upper airway resistance (RUA). The latter was also divided into nasal resistance (Rnas) and laryngeal plus extrathoracic tracheal resistance (Retr + lar). Furthermore, the inspiratory and expiratory components of each of these R values were estimated. Levels of RL, RLA and RUA were increased significantly during exercise but, during the recovery period, the values were significantly lower than those pre-exercise. RUA represented 82 per cent of RL at rest and this percentage did not change significantly during and after exercise. The nasal resistance to RUA ratio was always higher than 0.5. The fact that RL increased with exercise intensity was due to the increase of RUA during inspiration, and mainly a result of the increase of RLA during expiration. At fast trot, RLA represented 5 and 50 per cent of RL during inspiration and expiration respectively. It was concluded that heavy exercise induces in ponies an increase of RL, one reason for which could be the partial collapse of the extrathoracic and intrathoracic airways during inspiration and expiration respectively.     

Influence of exercise and joint topography on depth‐related spatial distribution of proteoglycan and collagen content in immature equine articular cartilage

Reasons for performing study: There is ample evidence on topographical heterogeneity of the principal biochemical components of articular cartilage over the surface of the joint and the influence of loading thereon, but no information on depth‐related zonal variation in horses. Objectives: To study depth‐related zonal variation in proteoglycan (PG) and collagen content in equine articular cartilage. Methods: Two techniques (safranin‐O densitometry and Fourier transform infrared spectroscopy) were applied to sections of articular cartilage from the proximal phalangeal bone of the metacarpophalangeal joint of 18‐month‐old Thoroughbreds that had been raised at pasture from age 0–18 months without (PASTEX) and with (CONDEX) additional exercise. Two sites were investigated: site 1 at the joint margin that is unloaded at rest or at slow gaits, but subjected to high‐intensity loading during athletic activity; and site 2, a continuously, but less intensively, loaded site in the centre of the joint. Results: Proteoglycan values increased from the surface to the deep layers of the cartilage, collagen content showed a reverse pattern. PG content was significantly higher at site 2 in both PASTEX and CONDEX animals without an effect of exercise. In the PASTEX animals collagen content was significantly higher at site 1, but in the CONDEX group the situation was reversed, due to a significant exercise effect on site 1, leading to a reduced collagen content. Conclusions: Collagen and PG content gradients agree with findings in other species. The observations on PG levels suggest that the exercise level was not strenuous. The collagen results in the PASTEX group confirmed earlier findings, the lower levels at site 1 in the CONDEX group being possibly due to an advancement of the physiological maturation process of collagen remodelling. Potential relevance: This study confirms earlier observations that even moderate variations in exercise level in early age may have significant effects on the collagen network of articular cartilage.     

Effect of dietary starch, fat, and bicarbonate content on exercise responses and serum creatine kinase activity in equine recurrent exertional rhabdomyolysis

To determine the effect of dietary starch, bicarbonate, and fat content on metabolic responses and serum creatine kinase (CK) activity in exercising Thoroughbreds with recurrent exertional rhabdomyolysis (RER), 5 RER horses were fed 3 isocaloric diets (28.8 Mcal/d [120.5 MJ/d]) for 3 weeks in a crossover design and exercised for 30 minutes on a treadmill 5 days/wk. On the last day of each diet, an incremental standardized exercise test (SET) was performed. The starch diet contained 40% digestible energy (DE) as starch and 5% as fat: the bicarbonate-starch diet was identical but was supplemented with sodium bicarbonate (4.2% of the pellet): and the fat diet provided 7% DE as starch and 20% as fat. Serum CK activity before the SET was similar among the diets. Serum CK activity (log transformed) after submaximal exercise differed dramatically among the diets and was greatest on the bicarbonate-starch diet (6.51 +/- 1.5) and lowest on the fat diet (5.71 +/- 0.6). Appreciable differences were observed in the severity of RER among individual horses. Postexercise plasma pH, bicarbonate concentration, and lactate concentration did not differ among the diets. Resting heart rates before the SET were markedly lower on the fat diet than on the starch diet. Muscle lactate and glycogen concentrations before and after the SET did not differ markedly among the diets. A high-fat, low-starch diet results in dramatically lower postexercise CK activity in severely affected RER horses than does a low-fat, high-starch diet without measurably altering muscle lactate and glycogen concentrations. Dietary bicarbonate supplementation at the concentration administered in this study did not prevent increased serum CK activity on a high-starch diet.     

The effect of exercise and conditioning on equine red blood cell characteristics

The effect of exercise and conditioning on 2,3-diphosphoglycerate levels was studied in nine mature horses. During a 12 minute exercise bout producing heart rates of 165 bpm, 2,3-DPG was significantly increased (p<.05). In addition, exercising levels of 2,3-DPG were increased (p<.05) approximately 8% after a six-week submaximal conditioning program. These increases could not be entirely attributed to changes in erythrocyte number. Mean corpuscular volume was also increased during exercise (p<.05) but was not altered by conditioning.