Mitochondrial myopathy in a german shepherd dog

A 9-month-old male German Shepherd dog was referred for evaluation of progressive exercise intolerance. Clinical examination revealed a stiff, stilted gait and marked atrophy and hypotonia of skeletal muscle. The dog had raised creatine kinase (181 U/liter), lactate dehydrogenase (510 U/liter), and aspartate aminotransferase (123.6 U/liter) levels, suggesting a muscle disease. Histochemical evaluation of muscle biopsies revealed the presence of subsarcolemmal oxidative activity, reduced nicotinamide adenine dinucleotide, and succinate dehydrogenase, and the absence of cytochrome oxidase activity. Ragged red fibers were demonstrated with Gomori trichrome stain. Ultrastructural examination of the muscle confirmed the presence of subsarcolemmal accumulations of mitochondria and morphologically atypical mitochondria.     

Lateral musculature in the whitemouth croaker (Micropogonias furnieri): its characterization with respect to different gonadal conditions

Histochemical and ultrastructural studies were performed on the lateral musculature from individual female whitemouth croaker, Micropogonias furnieri, at the anterior, medium and posterior regions. Based upon histochemical myosin-ATPase (m-ATPase) determination, diverse types of red, pink and white fibres were discerned. Red muscle had abundant mitochondria and stained intensely for aerobic enzymes, white muscle scarcely stained for the same enzymes and pink muscle responded in an intermediate manner. Both white and pink muscle had few mitochondria. The relative proportion of red muscle increased towards the caudal region; pink muscle diminished towards this region and white muscle modified its proportion only in the anterior region. m-ATPase activity showed differences in relation to the gonadal condition along the body, particularly in the white fibres at the anterior and medium regions.     

Episodic weakness associated with exertional lactic acidosis and myopathy in Old English sheepdog littermates.

Two Old English Sheepdog littermates were evaluated for weakness that developed during periods of minimally intense exercise. Lactic acidosis accompanied by increased muscle enzyme activity, an increased lactate/pyruvate ratio, and increased venous PO2 supported the possibility of defective mitochondrial oxygen use. Electromyographic abnormalities included increased insertional activity and complex repetitive discharges. Muscle alterations included scattered myofiber necrosis, abundant endomysial connective tissue, excessive glycogen accumulation, and greater than normal numbers and vacuolation of mitochondria. A distinctive pattern of subsarcolemmal mitochondrial aggregates, referred to as "ragged red fibers" in human mitochondrial myopathies, was observed in muscle biopsy samples from 1 dog. Several features of the disease in these dogs, including onset of weakness during early life, simultaneous disease in littermates, subtle nonprogressive weakness of at least 3 years' duration, and partial reversibility of lactic acidosis following rest were suggestive of an inborn error of metabolism, consistent with mitochondrial myopathy.     

Energy metabolism in relation to skeletal muscle fibre properties during treadmill exercise

The skeletal muscle metabolism of horses with a wide range of cardiocirculatory capacities was studied during a standardised near-maximal exercise test in relation to muscle fibre composition. Although the same amount of work was performed by all horses the amount of triglycerides and glycogen utilised and lactate accumulated differed widely. Both blood and muscle lactate accumulation were positively related to the amount of glycogen utilised. These factors were in turn positively associated with the percentage of Type IIB fibres and consequently negatively associated with the percentage of Type IIA fibres. The percentage of Type IIB fibres was negatively correlated to the activities of the oxidative enzymes citrate synthase and 3-hydroxyacyl-CoA-dehydrogenase and positively associated with the lactate dehydrogenase activity. The results indicate that the horses with a high proportion of Type IIB fibres had a lower muscle oxidative capacity and were, therefore, for energy production, more dependent upon glycolysis with subsequent lactate accumulation than horses which had a high proportion of Type IIA fibres. The different metabolic properties of the muscle fibre types indicate that the pattern of muscle fibre recruitment could have a significant effect on intramuscular substrate utilisation.     

Metabolic response to racing and fiber properties of skeletal muscle in standardbred and thoroughbred horses

Muscle biopsies were obtained from 17 Standardbred trotters (S) and 22 Thoroughbred racehorses (T) after races which varied in length from 1200 to 2840 m. Biopsies were taken between 2 and 14 minutes from the conclusion of the race. Histochemical and biochemical analyses were performed on all muscle samples. Standardbred horses possessed fewer Type II fibers, especially Type IIB fibers, and demonstrated a higher oxidative and lower glycolytic capacity in muscle than Thoroughbred horses. Intramuscular glycogen and lac-tate concentrations were lower in Standardbred horses after racing (mean 420 and 82 mmol/kg d.w. respectively) com-pared to Thoroughbred horses (mean 545 and 148 mmol/kg d.w. respectively). Low levels of intramuscular adenosine triphosphate (ATP) (mean S 17, T 15 mmol/kg d.w.) and creatine phosphate (mean S 28, T 42 mmol/kg d.w.) were recorded in both breeds after racing. Differences in muscle enzyme activities and metabolic response to exercise between breeds were related to the different muscle fiber compositions of Standardbred and Thoroughbred horses. Within both breeds, a high percentage of Type IIB fibers was associated with a low oxidative and high glycolytic capacity in muscle and a greater lactate accumulation after racing. In Standardbred horses low intramuscular ATP concentrations post-race were associated with a high percentage of Type IIB fibers and high lactate levels. No relationship was found between placing or speed of racing and intramuscular lactate and ATP concentrations post-race. This may have been due to many other factors which affected the outcome of a race and also due to the fact that it was not known when during a race lactate accumulation and ATP depletion occurred. Since the properties of fiber types appeared to be very different, the metabolic response within individual fiber types may be more closely related to fatigue than the metabolic response measured in whole muscle.     

Central core myopathy in a great dane

An eight-month-old female great dane was referred for investigation of exercise intolerance that had developed progressively over one month. Examination revealed poor muscle mass, elevated plasma levels of muscle-associated enzymes and electromyographic abnormalities. The dog was euthanased at 15 months old because of severe exercise intolerance. Tissue obtained by biopsy and at post mortem examination revealed prominent well defined central core structures within most muscle fibres, but no other abnormalities. The condition was diagnosed as a central core myopathy.     

Leiomyoma of a kidney in a dog: a rare diagnosis

A five-year-old, neutered male, Bernese Mountain Dog was referred because of a two-week history of hematuria and a one-week history of polydipsia and polyuria. Palpation of the dorsomedial mesogastrium revealed a mass on the left side, which was seen via radiography and ultrasonography to be attached to the left kidney. The left kidney and the attached mass were excised surgically, and the dog had an uneventful recovery. Histological examination of the mass revealed a dense proliferation of non-polymorphic spindle cells, which stained positive for smooth muscle actin via immunohistochemistry. Leiomyoma of the kidney was diagnosed. Postoperatively, there was mild azotaemia, which did not affect the general condition of the dog and was still present 15 months after surgery. Leiomyoma of the kidney is a benign tumor, and hematuria may be the only clinical sign.     

Estudio Histoquímico y Morfométrico de la Población Fibrilar del Músculo Diafragma. Parte I: Caballo y Perro

Histochemical and morphometric study of fibrilar population in diaphragm muscle. Part I: Horse and Dog By using m-ATPase techniques, under alkaline and acid pre-incubations, as well as metabolic techniques (α-MGPDH and NADH-TR), sections from the lumbar and costal portions of diaphragm muscle in horse and dog were analysed. Fibre samples were exposed to image-analysis observations in order to determine the lowest fibre diameter and obtain estadistical data. The findings revealed that the lumbar portion of horse diaphragm consists of fibre types I, IIA and IIB; in the costal portion, the fibre types I and IIA were present. In the dog, the lumbar and costal portions consisted of fibre types I, IIA and IIC, which showed some specific histochemical characteristics. Some data concerning both morphometrical and statistical fibre-type analysis are presented.     

Portal vein and aortic thromboses in a Siberian husky with ehrlichiosis and hypothyroidism

A six-year-old, neutered male Siberian husky was presented for euthanasia for end-stage liver disease. Examination of the dog raised questions regarding the severity of the condition. It had presented to the referring veterinarian with polyuria, polydipsia and weight loss. Blood tests at that time revealed elevated liver enzymes and hypoalbuminaemia. Cirrhosis was presumptively diagnosed, based on an ultrasound examination, which showed ascites with a normal liver. The dog had a history of hypothyroidism, which was controlled with levothyroxine. Physical examination revealed cachexia. A second abdominal ultrasound examination was performed and revealed portal vein and aortic thromboses. Tick titres showed a positive Ehrlichia canis titre of 1:640. Skin biopsies showed lymphoplasmacytic vasculitis. Doxycycline and aspirin treatment was initiated, and the clinical signs resolved. The authors concluded that the thrombi were primarily caused by chronic ehrlichiosis, based on the clinical findings and the response to treatment.     

Long-term survival after surgical excision of a schwannoma of the sixth cervical spinal nerve in a dog

A schwannoma of the sixth cervical spinal nerve in a 5-year-old Shetland Sheepdog was surgically excised, sparing the thoracic limb and resulting in long-term survival. The dog had been referred because of slowly progressive left thoracic limb lameness. The lesion was localized to the left suprascapular and musculocutaneous nerves or the C6 and C7 spinal nerves on the basis of neurologic examination, electrodiagnostic examination, and myelography. Surgical exploration revealed a mass, which was excised and identified histologically as a schwannoma. Three and a half years later, the dog was normal except for mild gait abnormality, focal muscle atrophy, and a focal area of decreased cutaneous sensation of the left thoracic limb.     

Adenocarcinoma metastasis of the intertransversarius cervicis muscle eliciting a right forelimb lameness in a dog

This article describes identification of a metastatic adenocarcinoma to the intertransversarius cervicis muscle using magnetic resonance imaging (MRI) in a dog that presented with chronic lameness of the right forelimb. Magnetic resonance imaging revealed a right sided, ovoid signal abnormality within the intertransversarius cervicis muscle lateral to the sixth cervical (C6) vertebra. The lesion was uniform, hyperintense on T2- and isointense on T1-weighted images to muscle and exhibited uniform contrast enhancement on T1-weighted images. The MRI findings were consistent with a neoplasia. Surgical excision was performed. Histopathological diagnosis was metastatic fibrous adenocarcinoma. The dog recovered rapidly but 6 months post-operatively he was killed because of lung metastases. Necropsy was declined and the primary tumour could not be identified.     

Morphological alterations in oxidative muscles and mitochondrial structure associated with equine atypical myopathy

REASONS FOR PERFORMING STUDY: There is a lack of well documented studies about muscular lesions in equine atypical myopathy (EAM). OBJECTIVES: To characterise morphopathological changes of striated muscles and myocardium, to progress understanding of this disease. METHODS: Thirty-two horses age 0.5-7 years kept on pasture were referred for a sudden ataxia/myoglobinuria syndrome. Clinical examination (stiffness, muscle pain, muscle fasciculations, abnormal gait, recumbency, myoglobinuria, tachycardia, sweating) and plasma CPK, LDH and AST levels were consistent with extensive myonecrosis and, together with anamnestic data, with so-called 'equine atypical myopathy' (EAM), a disease of unknown aetiology reported since 1939. Macroscopic and microscopic (histology, histoenzymology, ultrastructure) lesions were evaluated. RESULTS: Necropsic examination revealed large areas of muscle necrosis, the extent and severity of which varied between cases and muscles, but which were clearly more constant and severe in respiratory and postural muscles and in the myocardium. Histology highlighted a multifocal and monophasic process compatible with Zenker degeneration/necrosis that mostly and segmentally affected type 1 fibres. Histochemical evaluation revealed a weak and disorganised pattern of NADH tetrazolium reductase staining, the absence of calcium salts precipitates and a dramatic accumulation of lipid droplets. Ultrastructural examination often revealed fibres of which the sole modifications were altered mitochondria and sarcoplasmic lipidosis. CONCLUSIONS: Taken together, the data suggest that a primary alteration of mitochondria should be considered, although secondary mitochondrial abnormalities have yet to be ruled out. POTENTIAL RELEVANCE: The morphological features gathered here reveal that EAM shares most of the characteristics of toxic myopathies.     

Neuronal nitric oxide synthase is heterogeneously distributed in equine myofibers and highly expressed in endurance trained horses

Mammalian skeletal muscle expresses splice variants of neuronal nitric oxide synthase (nNOS). Skeletal muscles have a metabolically heterogeneous population of myofibers, and fiber composition in equine skeletal muscle is correlated with athletic ability in endurance events. In this study, we investigated whether nNOS expression in equine skeletal muscle is related to fiber type and endurance training. Biopsy samples obtained from the gluteus medius of sedentary- (SH) and endurance-trained (TH) horses were examined for the electrophoretic mobility of myosin heavy chain (MHC) and NOS activity. Serial tissue cross-sections were stained for myosin ATPase and nicotinamide adenine dinucleotide (NADH) reductase, and also immunostained for nNOS. The gluteus medius of TH had higher levels of nNOS expression and activity when compared to muscle from SH. In SH, nNOS was restricted to the subsarcolemmal area while in TH nNOS was also present at cytoplasmic sites. A splice variant of nNOS was heterogeneously distributed among the different myofibers, its expression being higher in fast-oxidative-glycolytic type IIA fibers than in fast-glycolytic type IIX fibers and absent in slow-twitch type I fibers. Trained horses had a significantly higher relative content of type IIA fibers, a greater oxidative capacity, and a lower percentage of type IIX fibers when compared with SH. The differences in muscle fiber typing between the 2 groups of horses reflected alterations that probably resulted from the endurance-training program. Overall, these results show that nNOS is differentially expressed and localized in the gluteus medius according to the fiber type and the athletic conditioning of the horses.     

Radiotherapy-induced tumor lysis syndrome in a dog with thymoma

A 13-year-old, female, mixed-breed dog with a huge cranial mediastinal mass underwent radiotherapy (RT). On the following day, the dog presented with lethargy and anorexia. Hematological examination revealed elevated levels of blood urea nitrogen, creatinine, inorganic phosphorus, potassium, lactate dehydrogenase, creatine phosphokinase and aspartate aminotransferase, decreased calcium level, and metabolic acidosis. Urine output markedly decreased. The patient recovered with fluid therapy and diuretic therapy; however, died suddenly from an unknown cause 11 days after RT completion. Histopathological examination after necropsy showed thymoma in the cranial mediastinum and extensive tubular necrosis of both kidneys which may be due to RT-induced tumor lysis syndrome (TLS). This report suggests that the risk of TLS should be evaluated in dogs with thymoma who undergo RT.     

Comparison of plasma metabolite concentrations and enzyme activities in beef cattle raised by different feeding systems in Korea, Japan and New Zealand

Concentrations of metabolites and immunoreactive insulin (IRI) and activities of enzymes related to energy metabolism were measured in plasma of Korean and Japanese beef cattle, which were raised by the indoor feeding system programmed to feed larger amount of roughage in their growing periods and larger amount of concentrate diet in their finishing periods (Japanese feeding system), and grazing New Zealand beef cattle. By the Japanese beef grading system, Korean and Japanese beef cattle showed high beef quality score, average grade 3.3 and 3.6, respectively. The plasma free fatty acid and lactate concentrations and lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and aspartate aminotransferase (AST) activities in Korean beef cattle were significantly higher than those in Japanese beef cattle. The plasma lactate concentration in Korean beef cattle was 8.40 mmol/l, which was similar to the values observed in lactic acidosis. The higher activities of plasma LDH, MDH and AST may indicate slight liver damage by slightly acidotic conditions in Korean beef cattle. New Zealand beef cattle fed on pasture which they harvest by grazing showed significantly lower plasma glucose, cholesterol, lactate and IRI concentrations and enzyme activities than those in Korean and Japanese beef cattle fed on larger amount of concentrate diets. Plasma metabolite concentrations and energy metabolism-related enzyme activities may be good indicators for evaluating metabolic conditions of beef cattle raised by different feeding systems.     

Caracteristicas Histoquimicas y Morfometricas de Algunos Musculos Extraoculares del Perro

Histochemical and Morphometric Aspects of some Extraocular Muscles of the Dog
This investigation was carried out on retractor bulbi, lateral and medial rectus muscles of six adult dogs. Tissues were collected from near the center of individual muscle bellies. These were stained for m-ATPase at varying pHs during preincubation, NADH-TR, Alpha-GPDH, Modified Mason trichrome and Hematoxyline and Eosin. Muscle fibers were classified as type I and type II, based on their reaction for m-ATPase. The retractor bulbi muscle was composed entirely of type II, oxidative, muscle fibers, with no glycolytic fibers. The rectus muscles presented a stratified composition, with superficial muscle fiber bundles containing a mixture of type II fibers oxidative and glycolytic, and central bundles of type I, mixed with type II. It was observed that there was less interstitial tissue in the center of the muscle bellies. Large diameter nerve fibers were also observed in the central layers.     

Enzyme histochemical features of equine gluteus muscle fibers

Gluteal muscle specimens were taken from 4 horses. From 1 of the 4 gluteal muscles, serial sections were prepared. Individual muscle fibers were identified and studied, using photomicrographs of sections stained by different enzyme histochemical methods. In specimens in which cytoplasmic soluble enzymes were studied, use was made of the semi-permeable membrane technique to hamper enzyme diffusion into reaction fluids. Enzymes involved in glycogenolysis, glycolysis, the tricarboxylic acid cycle, synthesis of reduced nicotinamide adenine dinucleotide phosphate, the pentose phosphate cycle, the alpha-glycerolphosphate shuttle, the respiratory chain, catabolism, and muscular contraction were studied. Some key enzymes of different metabolic pathways were also included. Each of 3 fiber types identified had distinct features. Type I fibers were characterized by a relatively strong aerobic capacity, compared with type IIA fibers, which were more glycolytic and had strong aerobic and moderate-to-strong anaerobic capacity. Type IIB fibers were characterized by a relatively low aerobic and a relatively high anaerobic capacity, and were glycolytic. Activities of phosphofructokinase, glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase, and alpha-naphtylesterase (nonspecific esterase) were so markedly different in the 3 fiber types that fiber typing was possible, aided by the demonstration of the activities of these enzymes. In type IIB fibers, the pentose phosphate cycle was more important than in the other fiber types. Except for the unexplained high alpha-naphtylesterase activity in type IIB fibers, catabolic enzymes were not active in healthy equine muscle fibers.     

Age-related changes in metabolic properties of equine skeletal muscle associated with muscle plasticity

The purpose of the present study was to determine the age-related changes in myosin heavy chain (MHC) composition and muscle oxidative and glycolytic capacity in 18 horses ranging in age from two to 30 years. Muscle samples were collected by excisional biopsy of the semimebranosus muscle. MHC expression and the key enzymatic activities were measured. There was no significant correlation between horse age and the proportions of type-IIA and type-IIX MHC isoforms. The percentage of type-I MHC isoforms decreased with advancing age. Muscle citrate synthase activity decreased, whereas lactate dehydrogenase activity increased with increasing age. Muscle 3-OH acyl CoA dehydrogenase activity did not change with ageing. The results suggest that, similar to humans, the oxidative capacity of equine skeletal muscle decreases with age. The age-related changes in muscle metabolic properties appear to be consistent with an age-related transition in MHC isoforms of equine skeletal muscle that shifts toward more glycolytic isoforms with age.     

Electron microscopic and histochemical studies of the transverse striated musculature of birds after long-term hypokinesia]

Electromicroscopical and histochemical studies of the gastrocnemius muscle has been carried out in 4 months old cockerels of the laying hybrid after hypokinesis lasting 15 and 30 days. It was found that restricted movement resulted in dystrophic changes of myofibrils, enlargement of the sarcoplasmic reticulum and oedem of interfibrillar spaces. Histochemical studies revealed focuses of increased activity of non-specific esterase decreased activity of dehydrogenase of lactic acid and a positive reaction of acid phosphatase.