Coagulology, biochemical profile and muscle pathology in calves diagnosed with nutritional muscular dystrophy.

The aim of this study was to explain the correlations between selenium deficiency, hemostatic and biochemical disorders, and the progression of pathological changes in calves diagnosed with nutritional muscular dystrophy (NMD). The study was performed on 20 calves with supplementation of 8 ml selenium and vitamin E preparation and 20 calves with symptoms of NMD. Blood was sampled from calves aged 5, 12 and 19 days. On day 19, samples of the biceps femoris muscle were collected from 6 animals in each group for histopathological analysis. The following blood parameters were determined: PLT, PT, TT, APTT, fibrinogen and D-dimer concentrations, antithrombin III activity, glucose, selenium and vitamin E concentrations, activity of CK, LDH and GSH-Px. Muscle sections were stained with H&E and HBFP. Platelet counts were significantly lower in calves with symptoms of NMD. No significant differences in coagulation parameters were observed between the groups. Sick calves were diagnosed with hyperglycemia and elevation of CK and LDH activity. Selenium and vitamin E concentrations in the blood serum were significantly lower in the experimental group together with significant drop in GSH-Px activity. Changes characteristic of Zenker's necrosis were observed in a muscle of the sick animals. To our best knowledge this is the first study in which the attempt was made to explain the relationship between selenium deficiency and changes in the coagulation system in ruminants.

Selenoproteins protect against avian nutritional muscular dystrophy by metabolizing peroxides and regulating redox/apoptotic signaling.

Nutritional muscular dystrophy (NMD) of chicks is induced by dietary selenium (Se)/vitamin E (Vit. E) deficiencies and may be associated with oxidative cell damage. To reveal the underlying mechanisms related to the presumed oxidative cell damage, we fed four groups of 1-day-old broiler chicks (n = 40/group) with a basal diet (BD; 10 µg Se/kg; no Vit. E added, -Se -Vit. E) or the BD plus all-rac-α-tocopheryl acetate at 50mg/kg (-Se +Vit. E), Se (as sodium selenite) at 0.3mg/kg (+Se -Vit. E), or both of these nutrients (+Se +Vit. E) for 6 weeks. High incidences of NMD (93%) and mortality (36%) of the chicks were induced by the BD, starting at week 3. Dietary Se deficiency alone also induced muscle fiber rupture and coagulation necrosis in the pectoral muscle of chicks at week 3 and thereafter, with increased (P < 0.05) malondialdehyde, decreased (P < 0.05) total antioxidant capacity, and diminished (P < 0.05) glutathione peroxidase activities in the muscle. To link these oxidative damages of the muscle cells to the Se-deficiency-induced NMD, we first determined gene expression of the potential 26 selenoproteins in the muscle of the chicks at week 2 before the onset of symptoms. Compared with the +Se chicks, the -Se chicks had lower (P < 0.05) muscle mRNA levels of Gpx1, Gpx3, Gpx4, Sepp1, Selo, Selk, Selu, Selh, Selm, Sepw1, and Sep15. The -Se chicks also had decreased (P < 0.05) production of 6 selenoproteins (long-form selenoprotein P (SelP-L), GPx1, GPx4, Sep15, SelW, and SelN), but increased levels (P < 0.05) of the short-form selenoprotein P in muscle at weeks 2 and 4. Dietary Se deficiency elevated (P < 0.05) muscle p53, cleaved caspase 3, cleaved caspase 9, cyclooxygenase 2 (COX2), focal adhesion kinase (FAK), phosphatidylinositol 3-kinase (PI3K), phospho-Akt, nuclear factor-κB (NF-κB), p38 mitogen-activated protein kinase (p38 MAPK), phospho-p38 MAPK, phospho-JNK, and phospho-ERK and decreased (P < 0.05) muscle procaspase 3, procaspase 9, and NF-κB inhibitor α. In conclusion, the downregulation of SelP-L, GPx1, GPx4, Sep15, SelW, and SelN by dietary Se deficiency might account for induced oxidative stress and the subsequent peroxidative damage of chick muscle cells via the activation of the p53/caspase 9/caspase 3, COX2/FAK/PI3K/Akt/NF-κB, and p38 MAPK/JNK/ERK signaling pathways. Metabolism of peroxides and redox regulation are likely to be the mechanisms whereby these selenoproteins prevented the onset of NMD in chicks.

Induction of eosinophilic enteritis and eosinophilia in rats by vitamin E and selenium deficiency.

Eosinophilic enteritis and eosinophilia, in addition to muscular dystrophy and occasionally liver necrosis, were experimentally induced in male Sprague-Dawley rats with a vitamin E- and selenium-deficient diet (basal diet) for 9 weeks. Cecum and ileum were affected more frequently and severely than other segments of the gastrointestinal tract. Eosinophils were multifocally or diffusely distributed in the intestinal wall but were most severe in the muscular layer and in the submucosa. Eosinophils were also present in stomach, liver with massive hepatocellular necrosis, and skeletal muscle with marked myonecrosis. Eosinophilic enteritis and eosinophilia were not observed in rats fed the basal diet supplemented with either vitamin E (100 or 200 ppm) or selenium (0.1 or 1.0 ppm). Eosinophilic enteritis, eosinophilia, and muscular dystrophy regressed when vitamin E- and selenium-deficient rats were subsequently fed either the vitamin E- or selenium-supplemented diet for 4-5 weeks. These findings suggest that vitamin E and selenium deficiency may play a role in the development of a diffuse type of eosinophilic enteritis and eosinophilia.

[Clinical and biochemical response in the prevention of nutritional myodystrophy in heifers]. / Klinická a biochemická odozva prevencie nutricnej myodystrofie u jalovíc.

Clinical and biochemical responses were studied after taking the measures to prevent nutrition muscular dystrophy in young cattle in the given ecological conditions. Analyzing the biological material (blood, hair, feed, soil), we found the sufficiently high saturation of heifer organisms with the microelement selenium and on the contrary, vitamin E deficiency. Sensitive indicators of the break-down of muscular tissue were the enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), and mainly creatinine kinase (CPK): the activities of these enzymes increased significantly after the heifers had been driven to pasture. The stay of animals in the run to get them used to the physical load before going to the pasture was not found to be a sufficient measure to prevent muscular nutrition myodystrophy if the animals had not been administered vitamin E and selenium supplements. Of the one hundred heifers we examined, seven animals began to show the signs of nutrition muscular dystrophy; none of these animals had been administered vitamin E and selenium supplements.

Swedish views on selenium.

For a long time selenium was known only for its toxic characteristics. During the last few decades selenium deficiency has been recognized as an important agricultural problem in Sweden. On average, grains and pastures only contain one tenth of the amount of selenium considered necessary to avoid symptoms of deficiency. However, the incidence of muscle degeneration among the animals has been low, probably due to imported animal feed. Since 1980 selenium has been added to animal feed, but only minor changes in the selenium content of Swedish food stuffs have been recorded. Some studies have shown that the average Swedish dietary intake of selenium is 10-70 micrograms/day. The lowest levels were found in vegan diets, based on locally-grown products. Swedish studies have found depressed glutathione peroxidase (GSH-Px) levels in patients with various skin disorders, including acne; myotonic dystrophy and rheumatic arthritis. Clinical trials with selenium supplementation have given positive results in some of these disorders as well as in some patients with disabling muscular and joint pains. Today there is a large sale of selenium tablets sometimes with additional vitamin E. The marketing of these tablets, however, is often based on unsubstantiated claims.

A preliminary study on selenium content of forages and local by-products in the Tadla area (Morocco) in connection with ovine nutritional myopathy.

In order to provide epidemiological support to tentative diagnosis of ovine enzootic myopathy in the Tadla area (Morocco), 20 feedstuff samples, representative of ovine rations, were tested for selenium content. All but one samples had a far lower value than the recommended level of 0.1 mg/kg dry matter.

[Nutritional muscular dystrophy in a clutch of ostrich chicks]. / Vlekspiersiekte in 'n broeisel volstruiskuikens.

Six 1 1/2-month old ostrich chickens in the Upington district of the Cape Province developed lameness. Two died and pathological examination of one of them revealed lesions identical to those of white muscle disease in the larger muscle groups. Vitamin E-selenium therapy cured the other 4. The diet of the animals consisted mainly of lucerne (alfalfa).

[Possibility of preventing muscular dystrophy in E avitaminosis with alpha-tocopheronolactone and diludin]. / Izuchenie vozmozhnosti preduprezhdeniia myshechnoi distrotii pri E-avitaminoze al'fa-tokoferonolaktonom i diludinom.

The efficacy of pharmacopeial alpha-tocopheryl acetate, alpha-tocopheronolactone and the antioxidant diludin was studied as to the possibilities of preventing E-hypovitaminosis muscle dystrophy in rabbits. alpha-Tocopheronolactone action was similar to that of pharmacopeial alpha-tocopheryl acetate as regards E-vitamin activity that was manifested by an increase in the body mass, decrease in the urine creatine index and maintenance of normal ubiquinone content, as well as of succinate-ubiquinone-reductase, NADH-ubiquinone-reductase and NADH-cytochrome c-reductase activity of rabbit skeletal muscle and heart mitochondria. On the contrary , as a result of the antioxidant administration, no direct correlation was found between ubiquinone content and activity of ubiquinone-dependent enzymatic systems of mitochondria. The data obtained attest to high E-vitamin activity of alpha-tocopheronolactone. The action of the latter was disclosed to be different from that produced by the antioxidant.

[Effectiveness of different alpha-tocopherol derivatives in preventing experimental muscular dystrophy in chickens]. / Izuchenie éffektivnosti razlichnykh proizvodnykh al'fa-tokoferola v preduprezhdenii éksperimental'noi myshechnoi distrofii u tsypliat.

Activity of vitamin E and its derivatives--alpha-tocopheryl quinone, its short chain analogue and alpha-tocopheryl acetate also containing short chain molecule were studied during chicken experimental muscular dystrophy. alpha-Tocopheryl quinone containing short chain molecule proved to be the most active substance; similarly to vitamin E the preparation increased the erythrocyte resistance to osmotic haemolysis, distinctly increased the content of ubiquinone and decreased concentration of ubichromenol in the chicken liver tissue as compared with E-deficient animals. At the same time, the content of ubiquinone was unaltered in skeletal muscles. The data obtained suggest that vitamin E could be effectively substituted by its derivatives for prevention an treatment of muscular dystrophy.

Ultrastructural studies of the myotendonous junction of selenium-deficient ducklings.

An ultrastructural study was made of the changes occurring within the gastrocnemius insertion of normal and selenium-deficient ducklings from 1 to 12 days of age. The cytologic characteristics of the fibroblasts, vessels, collagen, and muscle cells are described. Those exposed to the selenium deficiency showed major alterations of all components. The fibroblasts showed changes ranging from collapsed cisternae and degenerating mitochondria to rupture. The capillary endothelium was abnormal, as was the smooth muscle of arteriolar walls. The collagen sizes were altered, and the muscle cell termini showed major pathologic changes. The above alterations occurred within 4 days of exposure of the deficiency. The muscle cells of the body portion of the gastrocnemius showed no alterations until Day 8. The observations present evidence that indicates that connective-tissue-vascular abnormalities precede myopathic changes in nutritionally induced dystrophy. The significance of these findings is discussed with respect to the etiology of nutritionally induced dystrophy.

Effects of selenium and vitamin E deficiencies on reproduction, growth, blood components, and tissue lesions in sheep fed purified diets.

Three 2 X 2 factorial experiments were conducted with sheep fed purified diets to determine the effects of selenium and vitamin E on the incidence of white muscle disease (WMD) and blood components. All lambs reaching 6 weeks of age in the group receiving no vitamin E or selenium developed WMD lesions, whereas only a few lambs in either the +E - Se or -E + Se treatment groups developed these lesions. Plasma activities of creatine phosphokinase, lactic dehydrogenase and glutamic oxaloacetic transaminase were significantly elevated in lambs receiving no vitamin E or selenium, whereas these enzyme activities in those receiving only selenium were non-significantly elevated. The enzyme activities in plasma of those on the +E - Se or +E + Se treatments were maintained at low levels, suggesting vitamin E alone is more effective in preventing WMD than selenium alone. The metabolic interactions of these essentials are discussed.

Effects of selenium and vitamin E on blood selenium levels, tissue glutathione peroxidase activities and white muscle disease in sheep fed purified or hay diets.

The effects of selenium and vitamin E on blood selenium levels and tissue glutathione peroxidase activities were determined in sheep fed purified and hay diets. A significant increase of blood levels of this element and tissue glutathione peroxidase activities was found in sheep given selenium as compared to those not receiving this element. Of the tissues examined, the highest glutathione peroxidase activity was found in the heart. Vitamin E had no influence on either the blood selenium levels or upon the tissue glutathione peroxidase activity. With hydrogen peroxide as the substrate, tissue glutathione peroxidase activity was not correlated with the incidence of white muscle disease. Evidence is presented to suggest that 0.1 ppm dietary selenium is not sufficient under some conditions to meet the physiological requirements for this element.

Selenium in ruminant nutrition: a review.

The early interest in selenium related primarily to its toxicity, but since 1957 the element has been recognized as a dietary essential. The dietary requirement for selenium by most species is about .1 ppm. Deficiencies of selenium in cattle and sheep have been confirmed under natural grazing conditions in many countries of the world. Overt signs of inadequacy such as white muscle disease (nutritional muscular dystrophy) occur primarily in young calves or lambs born to selenium deficient dams. Infertility has increased in ewes grazing pastures low in selenium. In general, signs of deficiency have not occurred in older animals such as finishing beef cattle and lactating dairy cows. Subclinical deficiencies of selenium are not determined easily, however, and thus an inadequacy of the element may be limiting maximum animal performance under certain circumstances of drylot feeding. The current nutritional status of ruminant animals in many geographical areas and involving various feeding programs with this element has not been established. The recent widespread deficiency problems with nonruminants suggest that such an assessment should be made. Concentration of selenium in tissue, particularly in the liver, has been used in establishing selenium status of the animal. With lambs glutathione peroxidase activity in certain tissues may be a more accurate indicator of selenium adequacy than is selenium content of the tissue. Supplemental sodium selenite and sodium selenate by either oral administration or parenteral injection have prevented clinical signs of selenium deficiency and animal losses in both ruminant and nonruminant animals. Heavy pellets containing elemental selenium for placement in the rumen have proved effective. In general, organic forms of selenium are absorbed more readily by animals than are inorganic compounds. The dietary requirements for selenium and its metabolism are influenced by many nutrient interrelationships, including its interactions with sulfur, lipids, vitamin E, proteins, amino acids, and several microelements. The Food and Drug Administration gave approval in 1974 for the oral administration of supplemental selenium as either sodium selenite or sodium selenate to certain classes of swine and poultry. Similar approval in the United States for ruminants will require additional information, particularly with regard to the influence of dietary intake on concentrations of selenium in tissue and milk in beef and dairy animals.