Pathology of experimental vitamin D deficiency in turkeys and the effects of various vitamin D supplements.

One-day-old poults fed a vitamin D3-deficient diet were examined for clinical, biochemical, and morphological changes at 14 days of age. Changes in these parameters were compared at 15.5 and 17 days of age after one of the following vitamin D-replacement therapies was provided: water-soluble vitamin-mineral packs that contained vitamin D3; vitamin D3 in the feed; or vitamin D, 1,25-dihydroxyvitamin D3 in the feed. The vitamin D3-deficient poults were lame and had significant decreases in weight gain and in longitudinal skeletal growth. None of the therapies alleviated all these changes, but clinical lameness subsided in poults provided 1,25-dihydroxyvitamin D3. Calcium concentrations were significantly improved by all therapies. Treatment with vitamin D3 in the feed and water significantly increased 25-hydroxyvitamin D3 concentrations in vitamin D-deficient poults. The growth plate zones, with the exception of the mineralized hypertrophied zone, were all increased in length and not modified by treatment. However, the mineralized hypertrophied zones in the 1,25-dihydroxy-vitamin D3 group and the group receiving vitamin D3 in the water were comparable to that in the controls on day 15.5.     

Ultrastructural and ultracytochemical cyclic changes in the bovine uterine tube (oviduct) epithelium.

Ultrastructural and ultracytochemical features of the uterine tube (oviduct) infundibulum were studied in 8 Hereford cows, which were slaughtered in pairs on days 1 (estrus), 3, 9 or 10, and 18 of the estrous cycle. Fibrous granules (60 to 80 nm), which are supposedly related to basal body replication, were observed in the apical cytoplasm of ciliated cells. Close association between basal bodies and fibrous granules was apparent, especially during the follicular phase. Cilia were observed throughout of estrous cycle, although degeneration of cilia was not observed at any phase of the cycle. Prominent, striated rootlets were observed during both the follicular and luteal phases of the cycle. Maximum secretory cell differentiation was apparent during the follicular phase, at which time these cells were characterized by having a well-developed, rough endoplasmic reticulum with dilated cisternae, numerous ribosomes, and secretory granules of varied size and density. A prominent feature of the secretory granules was their membranous structure, consisting of concentric lamellae of equal dimensions. During the luteal phase, cytoplasmic protrusions were prominent, and extruded nuclei along with other cytoplasmic organelles were present in the tubal lumen. The presence of a well-developed, rough endoplasmic reticulum and numerous secretory granules during the follicular phase indicates that secretory activity of the uterine tube infundibulum may be stimulated by estrogen. During estrus, the cytoplasm of the stromal cells displayed abundant, rough endoplasmic reticulum with dilated cisternae. The increased and extensively dilated rough endoplasmic reticulum at the time of estrus probably indicates increased protein synthesis by the stromal cells. The presence of adenosine triphosphatase activity on the membrane of cilia suggests that this enzyme is involved in energy-forming reactions related to the vigorous action of cilia. The presence of acid phosphatase activity on the cell membrane of the epithelium, microvilli, and secretory granules may indicate involvement in the secretory mechanism of the cell.     

Regional and Cyclic Variations in the Ultrastructural Features of Secretory Cells in the Oviductal Epithelium of the Chinese Meishan Pig

The Chinese Meishan pig is prolific breed and it is considered that this pig has a capacity with higher rates of embryonic survival. The oviductal secretory cells may affect the embryonic development and survival. The aim of the present study was to investigate the ultrastructural features of secretory cells in the various regions of the Chinese Meishan pig oviduct during the follicular and luteal phases of the oestrous cycle. In the ampullar secretory cells, numerous secretory granules with moderately electron-dense matrices were present in the supranuclear cytoplasm and exocytosis of secretory granules was observed. The number of secretory granules was dramatically reduced in the ampullar secretory cells in the luteal phase. During the follicular phase in the fimbrial epithelium, the secretory cells contained rough endoplasmic reticulum and Golgi apparatus, but most cells had few small granules. In the luteal phase, the secretory cells in the ampullar and fimbrial epithelia extended beyond the luminal border of the ciliated cells. In the isthmus, many granules were present in the cytoplasm of secretory cells throughout the oestrous cycle, but the number of secretory granules was reduced in the luteal phase. The cytomorphometric data revealed that the height of ciliated cells decreased substantially in the fimbriae and ampulla at the luteal phase, while that of non-ciliated cells was less affected. These results suggest that the drastic reduction of cell height of ciliated cells cause the extrusion of most secretory cells beyond the ciliated cells in the fimbriae and ampulla during the luteal phase. In summary, our ultrastructural observations of Chinese Meishan pig oviduct revealed marked cyclic changes in the ultrastructural features of secretory cells. In particular, the ultrastructural features and the numbers of secretory granules were distinctive for each particular segment. These findings should provide insight into the regional and cellular differences in functions of secretory cells of the Chinese Meishan pig oviduct.     

Ultrastructure of parathyroid glands of growing chickens with vitamin D deficiency rickets

Ultrastructural studies were conducted on the parathyroid glands (PTG) of 2- to 7-week-old nontreated control growing broiler chickens (group A, 6 chickens); the same aged chickens with experimentally induced vitamin D deficiency rickets (18 morbid chickens each from groups B, C, and D); and in 4- to 9-week-old chickens naturally affected with rickets (group E, 8 chickens). Four types of principal cells were classified in the PTG of clinically normal birds and in PTG of birds with secondary hyperparathyroidism as follows: Type I cell (small cell in the resting phase); type II cell (medium-sized cell with well-developed rough-surfaced endoplasmic reticulum in the synthesizing phase); type III cell (large cell with well-developed Golgi apparatus and cytoplasmic organelles in the packaging and secretory phase); and type IV cell (medium-sized cell with poor cytoplasmic organelles in the involuting phase). Many coated vesicles were observed in type II and type III cells. In young control chickens, principal cells of the PTG were considered in the resting phase; in older control chickens, there were increased numbers of cells in the synthesizing phase. In the vitamin D deficiency groups, principal cells were in the synthesizing and packaging and secretory phases. In birds with naturally occurring rickets, principal cells were in packaging and secretory phases, with a small number of cells in the involuting phase. The ultrastructural changes of the PTG from the initial to final phases in growing chickens with secondary hyperparathyroidism were demonstrated.     

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.     

Vitamin E concentrations in blood plasma of sheep and in sheep tissues after a single intraruminal or intraperitoneal administration of DL-alpha-tocopheryl acetate

Twenty-five yearling wethers, weighing 40 to 45 kg were used in a trial designed to compare the effect of the route of administration of vitamin E upon plasma and tissue vitamin E status. Five control sheep without vitamin E administration were killed at the beginning of the trial. Of the remaining 20 sheep, 10 were given DL-alpha-tocopheryl acetate intraruminally and 10 by intraperitoneal injection. Of these, 10 wethers were killed three days after dosing (five from each treatment, IR3 and IP3) and the remaining wethers were killed eight days after dosing (IR8 and IP8). Blood samples were taken throughout the trial from sheep on the IR8 and IP8 treatments. Samples of whole adrenal gland, heart, liver, kidney, brachiocephalicus muscle, lung, pancreas and spleen were taken from all sheep at slaughter and were analysed for their vitamin E content. The blood plasma results showed that the most important index of vitamin E bioavailability, the area under the plasma concentration versus time curve, was greater in the intraperitoneally than intraruminally dosed sheep. There was a higher concentration of vitamin E in the tissues from the intraperitoneal group than the intraruminal group three days after the intraperitoneal injections. The results suggest that the greatest responses in vitamin E concentration in plasma and the tissues were recorded in sheep following intraperitoneal rather than intraruminal dosing with DL-alpha-tocopheryl acetate.     

An assessment of the efficacy and safety of selenium and cobalt included in an anthelmintic for sheep

A trial was devised to assess whether the administration of selenium and cobalt together with the anthelmintic mebendazole (Ovitelmin S&C) was safe and could improve the supplies of selenium and cobalt for adult sheep fed a whole grain diet, low in both elements, which produced a steady decrease in blood glutathione peroxidase (GSHPx) and plasma vitamin B12 concentrations. Ovitelmin S&C, when given orally in a single dose as a suspension containing 0.34 mg selenium/ml, and 0.44 mg cobalt/ml (to provide 0.11 mg selenium and 0.15 mg cobalt/kg liveweight) significantly increased the GSHPx activity in blood. After a second dose given 28 days later the rate of change increased from 2.5 to 3.5 u/g haemoglobin/day. The responses in GSHPx were similar for a preparation which contained twice the concentration of selenium. Ovitelmin S&C increased the concentration of vitamin B12 in the plasma by about 1000 pg/ml for four to seven days after each dose and the increases were similar to those observed in sheep treated with an Ovitelmin preparation containing 45 times more cobalt (providing 6.7 mg cobalt/kg liveweight). After 63 days, liver vitamin B12 concentrations were 43 per cent higher in the cobalt treated than in the untreated groups (P less than 0.01) with no differences among the groups given cobalt. Neither adverse reactions nor signs of toxicity followed the administration of Ovitelmin S&C or Ovitelmin containing the higher concentrations of selenium and cobalt.     

Expression of Survivin,Bcl‐2 and Bax Proteins in the Domestic Cat (Felis catus) Endometrium During the Oestrus Cycle

Apoptosis has been shown to be an important regulator of endometrium function. To clarify the regulation of apoptosis in the cat endometrium during the normal oestrus cycle, the expressions of the apoptosis‐related proteins (Bcl‐2 and Bax) and their correlation to the inhibitor of apoptosis protein Survivin were analysed using immunohistochemistry. The TUNEL technique (TdT‐mediated dUTP nick end labelling) was also used to detect DNA fragmentation characteristic of apoptotic cells. The results demonstrated that TUNEL labelling is not effective for the detection of apoptosis in cat endometrium. Survivin was expressed in the luminal and glandular epithelial cells of cat endometrium during all phases of the oestrus cycle. Survivin was localized in both the cytoplasm and nuclei of superficial and deep uterine gland cells during the luteal phase, while only cytoplasmic staining was observed during the follicular and anoestrus phases. Bax immunoreactivity in the cytoplasm of luminal and glandular epithelial cells as well as the smooth muscle cells of blood vessels was weak in the anoestrus phase. Compared with anoestrus, the intensity of Bax immunostaining was moderate in the follicular phase and increased dramatically in the luteal phase. Bcl‐2 immunostaining in the cytoplasm of luminal and glandular epithelial cells was moderate in the anoestrus phase. During the early follicular phase, cytoplasmic Bcl‐2 immunostaining was detected mostly in glandular epithelial cells. In the mid‐follicular phase, in glands, the amount of Bcl‐2 protein increased progressively from the superficial to the deep layer. In contrast, the expression of Bcl‐2 decreased in the secretory phase, being very low or absent in the mid‐ and late luteal phases. The overall results suggest that Survivin, Bax and Bcl‐2 proteins may cooperatively contribute to cell apoptosis and cell proliferation in the cat uterus during the oestrus cycle.     

Histomorphology of the Proventriculus of three Species of Australian Passerines: Lichmera indistincta, Zosterops lateralis and Poephila guttata

Histomorphology of the proventriculi of nectarivorous, granivorous and omnivorous passerines was studied. The proventriculus consisted of mucosal, submucosal, muscularis and serosal layers. Proventricular wall was thickest in omnivore, thinnest in granivore and intermediate in nectarivore. The openings of mucosal glands had a single spiral-like fold of mucosa in the omnivorous Silvereye, 2–3 spirals in the granivorous Zebra finch and 4–5 spirals in the nectarivorous Brown honeyeater. The mucosal glands were arranged in a uniform row in the wall of the organ and opened individually via a primary duct to the lumen of the proventriculus. The surface epithelial cells of the tunica mucosa contained secretory cells and the proventricular glands contained endocrine, neck and oxynticopeptic cells. The ultrastructural features of the oxynticopeptic cells changed from the oral to the aboral portion of the gland. In the oral region, the cytoplasm presented numerous, smaller (600–900 nm) homogenously dense zymogen secretory vesicles and larger (0.8–2.3 μm) pale floccular, tubular, mucin-like secretory granules, few small mitochondria and RER while in the aboral portion of the gland, the cytoplasm presented numerous, large mitochondria with closely packed cristae, secondary lysosome and infolding of the basal and apical cell membrane. The tunica sub mucosa was thin with occasional large blood vessels. The tunica muscularis consisted of inner longitudinal, middle circular and outer longitudinal layers. The external tunica serosa contained large bundles of myelinated and unmyelinated axons that were possibly branches of the intestinal nerve. The structural adaptations of the proventriculi of these three species to their various diets are discussed.     

藏绵羊卵巢组织学及卵泡超微形态的观察

旨在通过观察藏绵羊卵泡、黄体的组织学特征及卵泡的超微形态,探讨其与生理功能的关系.本研究运用大体解剖、常规组织切片和H.E染色及透射电镜技术对藏绵羊卵巢卵泡和黄体的组织结构特点以及卵泡的超微形态进行观察和分析.结果发现,藏绵羊黄体期和卵泡期卵巢的宽度和厚度存在显著差异(P＜0.05),而重量和长度无显著差异(P＞0.0...     

Normal vitamin D receptor function with increased expression of 25-hydroxyvitamin D(3)-24-hydroxylase in Corriedale sheep with inherited rickets

A likely inherited disease with gross and microscopic features of rickets has been recognised in Corriedale sheep in New Zealand, and a defect in end-organ responsiveness to vitamin D has been proposed as a likely mechanism. The aim of the present study was to characterize the mode of inheritance and determine the disease mechanism. Breeding trials showed that the mode of inheritance was autosomal recessive. Serum chemistry testing using different methodology and studies in cultured skin fibroblasts did not support our previous hypothesis of a defect in end-organ responsiveness. The studies revealed normal serum 1,25-dihydroxyvitamin D concentrations, normal vitamin D receptor function, and the presence of 24-hydroxylase mRNA in cells from affected sheep, even without induction by 1,25-dihydroxyvitamin D₃. In addition, osteocalcin mRNA expression was similar in both affected and control sheep. It was concluded that increased expression of 25-hydroxyvitamin D₃-24-hydroxylase, the enzyme that breaks down vitamin D, may be involved in the pathogenesis of inherited rickets in Corriedale sheep, but its role requires further clarification.     

Vitamin C treatment of embryos,but not donor cells,improves the cloned embryonic development in sheep

Vitamin C is not only an antioxidant but also a regulator of epigenetic modifications that can enhance the activity of the ten-eleven translocation (TET) family dioxygenases and promote the oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC). Here, we investigated the effects of vitamin C in regulating DNA methylation in sheep somatic cells or embryos in an effort to improve the cloned embryo development. Vitamin C treatment of sheep foetal fibroblast cells significantly increased the 5hmC levels but did not affect the 5mC levels in cells. After nuclear transfer, vitamin C-treated donor cells could not support a higher blastocyst development rate than non-treated cells. Although combination of serum starvation and vitamin C treatment could induce significant 5mC decrease in donor cells, it failed to promote the development of resultant cloned embryos. When cloned embryos were directly treated with vitamin C, the pre-implantation development of embryos and the 5hmC levels in blastocysts were significantly improved. This beneficial role of vitamin C on embryo development was also observed in fertilized embryos. Our results suggest that vitamin C treatment of the embryos, but not the donor cells, can improve the development of cloned sheep embryos.     

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.     

Stability of vitamin E in blood and plasma from cattle, sheep, and pigs.

The stability of alpha-tocopherol concentrations in sheep, cattle, and pig blood and plasma stored at different temperatures was examined. For all species, the vitamin was stable for at least 6 days in plasma stored at -20 C, 4 C, and 25 C and in blood stored at 4 C and 25 C. For sheep and cattle, the vitamin was stable for at least 6 days in plasma stored at 37 C, but it was unstable in blood from all species stored at 37 C and in pig plasma stored at 37 C.     

The retention and efficacy of soluble-glass boluses for providing selenium, cobalt and copper to sheep

The efficacy and retention of prototype and commercial (Cosecure) soluble-glass boluses containing selenium, cobalt and copper and having a similar size, density and composition were evaluated in separate 12-month trials with sheep grazing low selenium pastures but with adequate cobalt and copper levels. In both trials, sheep confirmed by X-ray as containing a bolus grew at a significantly greater rate than control sheep. This was attributed to the correction of a selenium deficiency. Although liver copper, and liver and serum vitamin B12 levels were significantly greater in treated sheep than controls on some occasions, the differences were not as great or consistent as with blood selenium levels. The rate of bolus loss was high and in both trials, approximately 40% of the sheep had lost the bolus after 6 months. No prototype boluses were present after 12 months. Intact Cosecure boluses were recovered from 3 sheep out of 30 after 12 months whilst one animal contained a part bolus. The recovered boluses were approximately 45% lighter than when originally administered. Bolus loss did not appear to be due to complete dissolution. In sheep which had lost the bolus, blood selenium levels fell with a half-life of 43 +/- 10 days.     

Electron microscopic studies of estrogen-induced ciliogenesis and secretion in uterine tube of the gilt.

The time required for occurrence of estrogen-induced uterine tubal (oviductal) ciliogenesis and for differentiation of secretory cells was studied, utilizing electron microscopy procedures. Sixteen cycling gilts were ovariectomized; 3 to 4 months later, 12 principal gilts were each given subcutaneous injections of 17 beta-estradiol in 0.5 ml of corn oil at the rate of 200 mug/day, and 4 control gilts were given injections of corn oil only at the rate of 0.5 ml/day. Two principals each were killed on days 1, 2, 3, 4, 5, and 7 after start of treatment. The epithelial heights were low and completely atrophied 3 to 4 months after ovariectomy. Uterine tubal cilia were absent in all the control gilts. Cytologic changes were not seen in the atrophied epithelium of ovariectomized gilts 1 day after estradiol treatment, but definite proliferative elements consisting of an extensive fibrillar meshwork encrusted with granules (60 to 80 nm) were observed in close association with the nuclear envelope and in the apical cytoplasm after 2 days of estradiol treatment. By day 3, enlarged electron-opaque granules referred to as condensation forms, undergoing various stages of depletion, were closely associated with radially arranged procentrioles. These associations have been referred to as generative complexes. The presence of many generative complexes indicates that maximal production of basal bodies can be expected after 3 days of treatment with estradiol. The depletion of the condensation forms produced hollow spheres with thin walls as the procentrioles grew in length and assembled their microtubules. Enlarged mature-appearing basal bodies were abundant in the cytoplasm after 3 days of estradiol treatment. These bodies aligned themselves linearly along the luminal surface of the cell. Small ciliary buds were then formed above the cell surface, and ciliary filamentogenesis occurred in the bud. Motile cilia were observed on day 3, but cilia numbers increased markedly between day 4 and days 5 and 7. Procentrioles were generated from the diplosomal centriole after 2 days of estradiol treatment. These observations have provided evidence for both ancentriolar and centriolar basal body replication in the ciliated cells of uterine tube of the gilt. Maximal secretory cell differentiation occurred after 3 days of estradiol treatment. Hypertrophy of cytoplasmic organelles was evident on day 3, but the number of secretory granules and amount of rough endoplasmic reticulum increased markedly on days 5 and 7. Close association of secretory granules, Golgi apparatus, and endoplasmic reticulum was evident after estadiol treatment. These data indicate that both ciliated and secretory cells are sensitive to estrogen.     

A Lectin Histochemical Study of Gustatory (Von Ebner's) Glands of the Horse Tongue

In the present work, gustatory glands (von Ebner's glands) of the horse tongue were examined by means of five peroxidase-conjugated lectins (PNA, DBA, SBA, UEA I, WGA), with and without prior sialidase digestion, in order to investigate the presence and distribution of carbohydrate residues in secretory cells and duct cells. The most intense staining of secretory cells was observed with PNA after pre-treatment with neuraminidase. This indicates that the terminal trisaccharide sequence sialic acid- (α2→3, 6) galactosyl (β1→3) N-acetylgalactosamine is the most frequent oligosaccharide chain present in glycoproteins secreted by horse gustatory glands. Secretory cells also contained oligosaccharides with terminal α-N-acetylgalactosamine and N-acetylglucosamine, whereas fucose was found in only a few glandular cells. The apical cytoplasm of duct lining cells reacted with all the lectins except WGA.     

Nuclear volume of cells in the stratum granulosum and theca folliculi interna of the largest dominant and atretic tertiary follicles in sheep during the selection process]

According to available literary data, an increased nuclear volume of endocrine gland cells, various structures of the diencephalon and endocrine ovarial structures could be observed when the respective organs and structures were more active; a decreased nuclear volume was revealed when the activity of the respective organ or its structure was suppressed (Mess, 1962; Maracek and Arendarcik, 1976, 1978). Attention was paid to the karyometric analysis of follicular cells of the stratum granulosum (SG) and secretory cells of the theca folliculi interna (TFI) during selection of the dominant ovulatory follicle (DOF). The aim of this study was to determine differences in hte nuclear volumes of the granulosa follicular cells and the secretory cells of the inner theca of the DO1F in comparison with the largest atretic follicles (AF) as well as to verify the use of karyometric variance analysis for the evaluation of the selection process of the dominant ovulatory tertiary follicle. Histological sections were prepared from large DOF and AF from the ovaries of 18 sheep aged 3-5 years and held under standard conditions of a controlled herd. Control group I (n = 3) consisted of Wallachian sheep on day 15 of the sexual cycle (day 0). Excisions of ovaries from the animals of the experimental group (n = 3) were made on day 16 of the cycle after i.m. treatment with 120 micrograms GnRH 10 and 7 hours prior to sampling. Control group II (n = 3) consisted of Tsigai sheep at the 9th or 10th day of the oestrous cycle (hour 0) and three experimental groups (n = 3 each) consisted of Tsigai sheep 24, 48 and 72 hours after i.m. treatment with 125 micrograms cloprostenol. After excision the ovaries were fixed in neutral formaline, cut into 4 mm thick transverse segments and subjected to standard histological processing. After staining with Harris' HE the 5-7 microns thick sections were karyometrically evaluated at a magnification of 2200 x according to the method of Palkovits (1961) using the PC programme Karyotest 03 (Maracek et al., 1991). 200 cells were evaluated from each sample and both cell types; altogether 7200 cells were examined. GnRH treatment (Dirigestran inj. Spofa) increased the nuclear volume of follicular cells (SG) in the dominant ovulatory follicle in the process of selection (Tab. I, Fig. 1). The former, however, reduced the nuclear volume not only of the SG follicular cells of LAF (Tab. II, Fig. 2) but also of the secretory cells of both the DOF and AF theca interna (Tab. II, Figs. 3, 4). Cloprostenol treatment (Oestrophan inj. Spofa) affected the follicular cells of the granulosa in the dominant ovulatory follicle.(ABSTRACT TRUNCATED AT 400 WORDS)     

Relative contributions of diet and sunshine to the overall vitamin D status of the grazing ewe

Circulating concentrations of 25-hydroxyvitamin D2 (25OHD2) and 25-hydroxyvitamin D3 (25OHD3) provide, respectively, a measure of the contributions of the diet and sunshine to the overall vitamin D status. These indices were measured in grazing sheep over a 16 month period which included their first pregnancy. Seasonal fluctuations in 25OHD3 concentrations were very marked, demonstrating both the dominance of photobiosynthesis as a source of vitamin D in the summer and also the lack of an effective storage mechanism. The concentration of 25OHD2 was subject to much smaller fluctuations, but when young grass was being consumed it was significantly lower than when mature grass or hay was fed. Supplementation in winter maintained circulating concentrations at a satisfactory level and may be a wise precaution in practice.