Rickets associated with vitamin D deficiency in young sheep

An outbreak of rickets in sheep under a year old (hoggs) appeared clinically as stiffness and rotation of the carpal joints. Histological studies confirmed the diagnosis and biochemical analyses of blood demonstrated a primary vitamin D deficiency.     

Ultrastructure of thyroid C cells in sheep treated with vitamin D3.

Ultrastructural observation was performed in C cells of sheep injected intramuscularly with a dose of 2 million IU of vitamin D3 daily for 10 days, 20 days and 30 days, respectively. After treatment with vitamin D3, hyperplasia and hypertrophy of C cells were noticed mainly at the periphery of the thyroid follicles. Most of hypertrophied C cells degranulated conspicuously and contained many prosecretory granules near the well developed Golgi apparatus which were in the actively secreting and packaging phase of their secretory cycle. The other C cells had prominent lamellar arrays of rough-endoplasmic reticulum and aggregations of free ribosomes in the cytoplasm which were interpreted to be in the actively synthesizing phase of their secretory cycle. Atrophic C cells which contained degenerative organelles in the cytoplasm were occasionally observed among the hypertrophied C cells. The present ultrastructural findings clarified that C cells synthesize and secrete calcitonin continuously due to prolonged hypercalcemia induced by long-term administration of excessive doses of vitamin D3 in sheep.     

Plasma and milk concentrations of vitamin D3 and 25-hydroxy vitamin D3 following intravenous injection of vitamin D3 or 25-hydroxy vitamin D3.

Plasma levels of vitamin D3 or 25-hydroxyvitamin D3 in ewes after administration of a single massive intravenous dose of vitamin D3 (2 X 10(6) IU) or 25-hydroxy vitamin D3 (5 mg) were determined at zero, one, two, three, five, ten and 20 days postinjection. In six ewes injected with vitamin D3 conversion of vitamin D3 to 25-hydroxy vitamin D3 resulted in a six-fold increase in the plasma 25-hydroxy vitamin D3 level within one day. Elevated levels were maintained until day 10 but by day 20 a substantial decline in the plasma 25-hydroxy vitamin D3 level had occurred. Peak levels of vitamin D3 were reached one day after injection and then continuously declined until day 20. Administration of 25-hydroxy vitamin D3 increased plasma concentrations of 25-hydroxy vitamin D3 to fivefold higher levels than those observed when vitamin D3 was injected, with approximately threefold higher levels of 25-hydroxy vitamin D3 maintained for five days. On day 10 and day 20 ewes which were injected with 25-hydroxy vitamin D3 still maintained plasma levels of 25-hydroxy vitamin D3 which were twice as high as those of ewes injected with vitamin D3. In six ewes injected with vitamin D3, a sharp increase in vitamin D3 level in milk occurred within one day and more than a tenfold elevation of milk vitamin D3 concentrations were maintained for ten days. By 20 days the milk vitamin D3 level had returned to preinjection levels. These observations suggest that indirect supplementation of the suckling ruminant with vitamin D3 may be achieved through maternal injection and subsequent mammary transfer.     

Association of Klebsiella organisms with pulmonary lesions in sheep.

Small nodules were seen on chest wall and in the lungs of sheep slaughtered in Al-Ahsa abattoir, Saudi Arabia. Klebsiella pneumoniae subspecies ozaenae was isolated and characterized. The histopathology of the nodules was described.     

Distribution of lesions in fetal brains following experimental infection of pregnant sheep with Toxoplasma gondii

Six ovine fetal brains were harvested 33 to 35 days postchallenge from 5 ewes, each of which was given 3000 Toxoplasma gondii oocysts on day 90 of pregnancy. Histopathologic examination of transverse sections taken at 13 levels in the fetal brains revealed the presence of toxoplasmosis-related lesions in all 6 brains. However, lesions were not randomly distributed (P = .007); they were most numerous at the level of the optic tract, the rostral margin of the pons, and 4 mm caudal to the ansate sulcus and were absent in all sections at the level of the caudal cerebellum. Lesion distribution may be due to hemodynamic factors, differences in the expression of endothelial surface receptor molecules at the level of the blood-brain barrier, or the presence of localized permissive/inhibitory factors within the brain. The results have implications for the selection of areas of brain from aborted ovine fetuses to be examined histopathologically for laboratory diagnosis.     

Effect of parenteral vitamin D3 on plasma 25-hydroxyvitamin D3 concentration in sheep

Vitamin D3 ranging in total amount from 10(5) iu (2.5 mg) to 9 X 10(5) iu (22.5 mg) was given intramuscularly either as a single injection or in three aliquots at three-weekly intervals to housed nonpregnant ewes on a vitamin D deficient diet. The effects on plasma concentrations of 25-hydroxyvitamin D3 (25-OHD3), the major metabolite of vitamin D, were monitored. The increase in plasma 25-OHD3 showed a large variation between animals and was related to, but not proportional to, the dose. None of the treatments produced 25-OHD3 concentrations greater than those in grazing sheep in summer. Repeated dosing provided for more efficient use of the injected vitamin D3.     

Effects of gestation and lactation on vitamin D receptor amounts in goats and sheep

During gestation and lactation, an increased demand for calcium (Ca) due to the development of fetal skeleton and excretion via milk is observed. The higher need for Ca is met by an augmented mobilisation of Ca from bones and by an increased absorption from the intestines. The main influence on this physiological process of active absorption has Vitamin D, acting through Vitamin D receptors (VDR) located in the mucosal wall of the intestines, thus increasing Ca absorption. As a consequence of inadequate Ca absorption, metabolic diseases like milk fever can develop. In this study immunohistochemical procedures were applied to colon mucosa biopsies of pregnant and lactating goats and sheep, to study the effect of late gestation, parturition and lactation on VDR amount. Colon mucosa biopsies were collected 2 weeks before parturition, 1 and 4 weeks post partum (pp), 2, 3, 4, and 5 months pp from 11 dairy goats and 11 sheep. Immunohistochemistry was performed employing a biotinylated monoclonal rat anti-VDR antibody and streptavidin peroxidase techniques. Nuclei and cytoplasm of enterocytes stained positively for VDRs. Strongest immunoreactions were observed in intermediate and superficial glandular cells. The biopsy samples taken during early lactation revealed a lower immunoreaction for VDR compared with samples taken during later stages of lactation. In conclusion, immunochemistry and biopsy technology are useful tools to assess changes in VDR expression in relation to varying demands for Ca in the process of a reproductive cycle. These results show that in dairy goats and sheep, an influence of gestation and lactation on VDR is obvious.     

Effects in sheep of high dietary levels of vitamin D3 as measured by circulating concentrations of some vitamin D metabolites and skeletal examination

Housed non-pregnant ewes were given diets supplemented with five, 10, 20, 40 and 80 times the Agricultural Research Council's recommended requirement for vitamin D for 16 weeks. The effects on vitamin D status were assessed by measuring 25-hydroxylated vitamin D metabolites (25-OHD) in plasma at intervals. After 16 weeks, animals on the lowest and two highest vitamin D intakes were killed for histological examination of selected soft tissues and bones. Circulating concentrations of 25-OHD increased with intake and time during the experiment. However the increases were not proportional to intake and only when intakes exceeded 20 times the recommended requirement were concentrations greater than those seen in summer in grazing sheep. There was some evidence for the production of an additional vitamin D metabolite in animals on high vitamin D intakes. No abnormalities were detected in any of the tissues examined histologically.     

Providing vitamin D to confined sheep by oral supplementation vs ultraviolet irradiation

Serial vitamin D3 (D3) and 25-hydroxyvitamin D3 (25 OH D3) concentrations of plasma were measured in confined, shorn sheep that had either been supplemented with vitamin D3 (50 micrograms/d) or exposed daily to ultraviolet irradiation (UVI). In the sheep administered D3 orally, plasma D3 increased continuously until d 35. This was followed by small fluctuations of the plasma D3 concentrations until a plateau was reached after 56 d of supplementation (.94 ng/ml plasma). Plasma 25 OH D3 concentrations increased continuously and plateaued between d 65 to 75 at about 21 ng/ml plasma. In the UVI sheep, plasma D3 and 25 OH D3 concentrations increased continuously for the first 49 d, then plateaued at 2.03 ng D3 and 29.6 ng 25 OH D3/ml. When a plateau was reached in plasma 25 OH D3 concentrations in both treatment groups, a 3H-labeled tracer dose of 25 OH D3 was given i.v., and disappearance of the 3H-labeled 25 OH D3 was followed. The UVI group had a faster decline in specific activities during the first exponential phase but a slower decline during the prolonged terminal elimination phase. These differences are reflected in the intercompartmental transfer rates. Our data indicate that UVI is as effective as oral vitamin D3 supplementation for improving vitamin D status of confined sheep.