Effect of whole-body potassium depletion on plasma, erythrocyte, and middle gluteal muscle potassium concentration of healthy, adult horses.

The effects of whole-body potassium depletion induced by food deprivation on plasma, erythrocyte, and middle gluteal muscle K concentrations was quantified in 16 healthy, adult horses before, during, and at the end of a 7-day period of food deprivation during which water and sodium chloride were available ad libitum. Potassium concentrations were determined by atomic absorption spectroscopy. Plasma K concentration remained constant (3.49 +/- 0.09 mM K/L of plasma; mean +/- SEM) throughout the study. Erythrocyte potassium concentration decreased from 93.10 +/- 1.94 mM K/L of erythrocytes on day 0 to 88.63 +/- 2.39 mM K/L of erythrocytes on day 2 (decrease of 4.8%; P less than 0.05) and thereafter did not change. The K concentration of the middle gluteal muscle decreased from 91.06 +/- 2.96 microM K/g of muscle (wet weight) to 79.61 +/- 2.09 microM K/g of muscle (decrease of 12.6%; P less than 0.05) on day 4 and decreased further on day 7 to 73.62 +/- 1.85 microM K/g of muscle (decrease of 19.2%; P less than 0.05). There was no correlation between the plasma and erythrocyte K concentrations (r = -0.066), the erythrocyte and middle gluteal muscle K concentrations (r = 0.167), or the plasma and middle gluteal muscle potassium concentrations (r = -0.018). The water content of the middle gluteal muscle remained constant (73.23 +/- 0.36%) throughout the study. Erythrocyte membrane potential did not change (-99.26 +/- 0.87 mV) during the study, whereas the magnitude of the membrane potential of the middle gluteal muscle decreased from -105.84 +/- 1.67 mV on day 0 to -100.93 +/- 2.10 mV on day 7 (P less than 0.05).     

Electronic measurement of erythrocyte volume and volume heterogeneity in horses during erythrocyte regeneration associated with experimental anemias

Anemia was induced in three groups of horses by moderate or severe acute hemorrhage, or by acetyl phenylhydrazine-induced hemolysis (Groups I, II, and III, respectively). Serial hemograms were done on a multichannel automated blood cell counter with histogram capability. Changes in hematocrit, mean cell volume, erythrocyte number, red cell distribution width (RDW), and standard deviation of erythrocyte volume were examined over time. Significant increases in mean cell volume were first detectable by days 17, 20, and 14 and reached maximum by days 43, 41, and 29, in Groups I, II, and III, respectively (P less than 0.05). Increased mean cell volume was interpreted as reflecting accelerated erythrocyte regeneration; however, not all horses with accelerated regeneration had changes in mean cell volume. Estimated erythrocyte production rate correlated poorly with hematocrit nadir and change in mean cell volume (r = 0.37 and r = 0.36, respectively, P greater than 0.05). In some horses effective regeneration occurs without development of macrocytosis. Mean cell volume remained increased after other parameters returned to control values, suggesting that mean cell volume values may provide retrospective evidence of altered erythrocyte turnover. Anisocytosis as indicated by significant increases in the standard deviation was greatest during the early part of the regenerative response, reaching maximum values on days 30, 28, and 21 in Groups I, II, and III, respectively, and began to decrease as homogeneous repopulation with macrocytes occurred. Red cell distribution width increased significantly only in severe hemorrhage and hemolysis groups, reaching mean maximum values of 24.3 on day 20 and of 26.4 on day 21 in Groups II and III, respectively (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Glucose uptake in horses with polysaccharide storage myopathy

OBJECTIVE: To determine whether excessive glycogen accumulation in skeletal muscle of Quarter Horses with polysaccharide storage myopathy (PSSM) is a result of enhanced cellular uptake of glucose. ANIMALS: 6 horses with PSSM and 10 healthy (control) horses. PROCEDURE: Intravenous glucose tolerance tests (IVGTT), oral glucose tolerance tests (OGTT), and modified insulin tolerance tests (MITT) were performed. Plasma glucose and insulin concentrations were measured in blood samples collected before and for up to 8 hours after glucose or insulin administration. RESULTS: Peak glucose concentrations during IVGTT were similar for both groups of horses, but rate of glucose clearance was 1.5 times faster in horses with PSSM than in controls. Moreover, circulating concentrations of insulin before and after glucose injection were lower in the PSSM group. Blood glucose concentrations from minute 90 to minute 300 of the OGTT were lower in horses with PSSM than in controls. The MITT resulted in acute decreases in blood glucose concentrations in both groups of horses; however, horses with PSSM sustained low blood glucose concentrations for more than 3 hours after insulin injection, whereas blood glucose concentrations in controls returned to baseline values within 2 hours. CONCLUSIONS: Quarter Horses with PSSM have enhanced cellular uptake of glucose that may be, in part, caused by an increased sensitivity to insulin. CLINICAL RELEVANCE: Horses with PSSM have an increased rate of glucose clearance in response to insulin secretion. Thus, diets low in soluble carbohydrate may be the most effective way to decrease glycogen accumulation in skeletal muscle of these horses.     

Severe diaphragmatic necrosis in 4 horses with degenerative myopathy

Severe diaphragmatic necrosis occurred in horses with degenerative myopathy due to polysaccharide storage myopathy (n = 2), nutritional myopathy (n = 1), and vasculitis (n = 1). Blood gas analysis performed in 1 horse indicated development of respiratory acidosis. Respiratory muscle necrosis can be severe in horses with degenerative myopathy and can lead to respiratory failure.     

Insulin sensitivity in Belgian horses with polysaccharide storage myopathy

OBJECTIVE: To determine insulin sensitivity, proportions of muscle fiber types, and activities of glycogenolytic and glycolytic enzymes in Belgians with and without polysaccharide storage myopathy (PSSM). ANIMALS: 10 Quarter Horses (QHs) and 103 Belgians in which PSSM status had been determined. PROCEDURES: To determine insulin sensitivity, a hyperinsulinemic euglycemic clamp (HEC) technique was used in 5 Belgians with PSSM and 5 Belgians without PSSM. Insulin was infused i.v. at 3 mU/min/kg for 3 hours, and concentrations of blood glucose and plasma insulin were determined throughout. An i.v. infusion of glucose was administered to maintain blood glucose concentration at 100 mg/dL. Activities of glycogenolytic and glycolytic enzymes were assessed in snap-frozen biopsy specimens of gluteus medius muscle obtained from 4 Belgians with PSSM and 5 Belgians without PSSM. Percentages of type 1, 2a, and 2b muscle fibers were determined via evaluation of >or= 250 muscle fibers in biopsy specimens obtained from each Belgian used in the aforementioned studies and from 10 QHs (5 with PSSM and 5 without PSSM). RESULTS: Belgians with and without PSSM were not significantly different with respect to whole-body insulin sensitivity, muscle activities of glycogenolytic and glycolytic enzymes, or proportions of muscle fiber types. However, Belgians had an increased proportion of type 2a and decreased proportion of type 2b muscle fibers, compared with proportions in QHs, regardless of PSSM status. CONCLUSIONS AND CLINICAL RELEVANCE: PSSM in Belgians may be attributable to excessive glycogen synthesis rather than decreased glycogen utilization or enhanced glucose uptake into muscle cells.     

Ubiquitin expression in muscle from horses with polysaccharide storage myopathy

Serial sections of formalin-fixed, paraffin-embedded muscle biopsy specimens from 28 Quarter Horse, Paint, and draft-related breeds, aged 0.5-23 years, were treated with periodic acid-Schiff (PAS) stain for glycogen and were immunostained to detect ubiquitin expression. On the basis of findings in PAS-stained sections, a diagnosis of equine polysaccharide storage myopathy (EPSSM) was made in 22 horses aged 2-23 years (mean, 9.4 years); samples from 6 horses aged 0.5-15 years (mean, 7.3 years) had a normal PAS staining pattern, with no relevant lesions. Ubiquitin expression was detected in all but a 2-year-old EPSSM-affected horse and was not detected in the non-EPSSM-affected horses. Ubiquitin expression was greater than the degree of PAS-positive, amylase-resistant material, and ubiquitin was detected in aggregates of amylase-sensitive glycogen as well as in aggregates of amylase-resistant material. Results suggest that glycogen aggregates develop and are ubiquitinated prior to development of amylase-resistant inclusions. Ubiquitin immunostaining may be most useful for confirming the diagnosis of EPSSM in horses with only amylase-sensitive glycogen aggregates and in horses with early amylase-resistant inclusions. However, ubiquitin immunostaining is no more sensitive than is PAS staining for diagnosis of EPSSM.     

Postanesthetic hind limb adductor myopathy in five horses

Five horses that underwent prolonged anesthesia (greater than 3 hours) in dorsal recumbency for a surgical procedure were unable to stand after recovery and were euthanatized. A provisional diagnosis of postanesthetic myopathy was confirmed at necropsy in all 5 horses. However, distribution of affected muscles in these horses was atypical, because there was bilateral hind limb adductor muscle involvement.     

Polysaccharide storage myopathy in Cob Normand draft horses

Gluteus medius muscle was sampled from 53 Cob Normand horses for histologic evaluation. Twenty horses (38%) exhibited amylase-resistant material in myocytes consistent with polysaccharide storage myopathy. Diameter of affected type II fibers was increased (67.7 +/- 21.4 microm) compared with normal ones (57.3 +/- 19.7 microm). Two groups were distinguished by quantitative study. The first group (n = 14; 26%) was characterized by a low percentage of fibers (m = 0.98%) containing aggregates occurring singly or in perifascicular clusters without myopathic changes. The second group (n = 6; 11%) was characterized by a high percentage (m = 18.1%) of fibers containing aggregates scattered in biopsy with chronic myopathic changes. Re-biopsy of 4 horses showed an increase with time in the number of aggregate-containing fibers for horses of the first group only. In 1 necropsied horse, aggregates were observed in a wide range of muscles including smooth muscles. Ultrastructurally, granular material was found interspersed among arrays of filamentous material.     

MCT1, MCT4 and CD147 gene polymorphisms in healthy horses and horses with myopathy

Polymorphisms in human lactate transporter proteins (monocarboxylate transporters; MCTs), especially the MCT1 isoform, can affect lactate transport activity and cause signs of exercise-induced myopathy. Muscles express MCT1, MCT4 and CD147, an ancillary protein, indispensable for the activity of MCT1 and MCT4. We sequenced the coding sequence (cDNA) of horse MCT4 for the first time and examined polymorphisms in the cDNA of MCT1, MCT4 and CD147 of 16 healthy horses. To study whether signs of myopathy are linked to the polymorphisms, biopsy samples were taken from 26 horses with exercise-induced recurrent myopathy. Two polymorphisms that cause a change in amino acid sequence were found in MCT1 (Val₄₃₂Ile and Lys₄₅₇Gln) and one in CD147 (Met₁₂₅Val). All polymorphisms in MCT4 were silent. Mutations in MCT1 or CD147 in equine muscle were not associated with myopathy. In the future, a functional study design is needed to evaluate the physiological role of the polymorphisms found.     

MCT1, MCT4 and CD147 gene polymorphisms in healthy horses and horses with myopathy

Polymorphisms in human lactate transporter proteins (monocarboxylate transporters; MCTs), especially the MCT1 isoform, can affect lactate transport activity and cause signs of exercise-induced myopathy. Muscles express MCT1, MCT4 and CD147, an ancillary protein, indispensable for the activity of MCT1 and MCT4. We sequenced the coding sequence (cDNA) of horse MCT4 for the first time and examined polymorphisms in the cDNA of MCT1, MCT4 and CD147 of 16 healthy horses. To study whether signs of myopathy are linked to the polymorphisms, biopsy samples were taken from 26 horses with exercise-induced recurrent myopathy. Two polymorphisms that cause a change in amino acid sequence were found in MCT1 (Val₄₃₂Ile and Lys₄₅₇Gln) and one in CD147 (Met₁₂₅Val). All polymorphisms in MCT4 were silent. Mutations in MCT1 or CD147 in equine muscle were not associated with myopathy. In the future, a functional study design is needed to evaluate the physiological role of the polymorphisms found.     

Comparison of three methods for estimation of exercise-related ion losses in sweat of horses

OBJECTIVE: To quantify total fluid loss in sweat of Thoroughbreds during >3 hours of low-intensity exercise in controlled conditions and to calculate and compare estimated ion losses in sweat, according to 3 methods. ANIMALS: 6 exercise-trained Thoroughbreds. PROCEDURE: Fluid and ion losses in sweat were measured in 6 horses exercising at 40% of the speed that elicited maximum oxygen consumption for 45 km. Horses were given a 15-minute rest period at the end of three 15-km exercise phases. Horses completed 2 exercise trials. Ion losses in sweat were calculated, using measurements of local sweating rate and sweat ion composition (SWT), change in net exchangeable cation content (CAT), and change in extracellular ion content (PLAS) derived from plasma total solids and ion concentrations. RESULTS: Measurement of SWT revealed a mean (+/- SEM) fluid loss in sweat during 45 km of exercise of 27.5 +/- 1.6 L. Total ion loss in sweat was approximately 241 g or 7.8 mol with higher sodium losses in the second and third phases of exercise compared with the first phase. Losses of sodium and potassium calculated by SWT or CAT were not significantly different from each other, whereas losses of these ions as determined by PLAS were significantly lower. CONCLUSIONS AND CLINICAL RELEVANCE: Calculation of ion losses from a mean whole body sweating rate extrapolated from either local sweating rate and sweat ion composition or from change in net exchangeable cation content provide similar results, whereas ion losses determined by changes in extracellular ion content derived from plasma total solids and ion concentration results in underestimation of actual losses.     

Concurrent atypical myopathy and equine dysautonomia in two horses

This report concerns 2 horses that suffered typical clinical signs of atypical myopathy (AM) and equine grass sickness (EGS) concurrently. Clinical details and pathological lesions of the cases are described. EGS and AM are relatively rare diseases and the concurrency of the diseases in the same animals is therefore considered unlikely to be a coincidence. However, it is not suggested that the evidence shows a common aetiology but rather the existence of common predisposing causes.     

Atypical myoglobinuria: an acute myopathy in grazing horses

Four out of 12 horses grazing a field in Berkshire, England, suffered a prostrating illness and died within 12 to 72 h. Serum biochemical abnormalities, including markedly elevated muscle enzymes, were demonstrated and at post mortem widespread myodegeneration was found in both skeletal muscle and myocardium. Urine analysis revealed myoglobinuria, and renal changes were seen histologically. Although similar pathologically, the clinical syndrome and circumstances of the outbreak were not typical of equine exertional rhabdomyolysis (EER). The outbreak bore a striking resemblance to other reported sporadic outbreaks of an atypical myoglobinuria occurring in grazing horses. A number of potential aetiological and contributory factors (including herbicide toxicity) were considered, but the aetiology remains unresolved.     

Risk factors associated with colic in horses

Many factors have been identified as risk factors for colic in horses in several epidemiological studies. The aim of our paper was to review the results of 12 epidemiological studies, in order to assess the impact of each risk factor for colic. According to the literature, the factors that increase the risk of colic are feeding practices (type and quality of food, type and changes of feeding), the intrinsic factors of horses (sex, age and breed), management (type and changes of housing and activity), medical history (a previous colic, administration of a medical treatment) and parasite control (the presence of worms and type of deworming program). Several individual factors were incriminated as risk factors by all the studies. Nevertheless, the different studies did not always agree on the role of other risk factors. The conclusions were tightly related to several criteria in the selection of the study population, like the type of the epidemiological study, the number and the origin of horses included and the location of the study.