Expression of calbindin-D9k messenger ribonucleic acid in the gastrointestinal tract of dairy cattle

The calcium demands of pregnancy and lactation are known to up-regulate expression of Calbindin-D9k (CaBP-9k) mRNA in the intestines. The gastrointestinal CaBP-9k mRNA expressions has not been studied in dairy cows, which are bound to experience several pregnancies and lactation stages. In this study, the CaBP-9k mRNA expression were examined in the gastrointestinal tract of Holstein dairy cattle by Northern blot analysis. Detectable expression of CaBP-9k mRNA was localized in the proximal portion of the small intestines. These expressions were higher at the most proximal region of the duodenum and gradually decreased distally. The duodenal CaBP-9k mRNA was detected in all dairy cattle from 0.4 to 83.4 months old, but was not detectable in foetuses. There were no significant correlations between the age and the levels of CaBP-9k mRNA expression or between the plasma 1,25-(OH)2D3 concentrations and the levels of CaBP-9k mRNA expression.     

Effects of coumestrol administration to maternal mice during pregnancy and lactation on renal Ca metabolism in neonatal mice

The present study was conducted to clarify the effects of coumestrol administration to maternal mice during pregnancy and lactation on serum Ca and Ca metabolism in their neonatal mice. From 6.5 to 16.5 days post coitus and from 3 to 10 days after parturition, maternal mice were administered at 200 µg/kg body weight/day of coumestrol. Coumestrol administration did not affect weight gains, serum Ca and the expression of vitamin D receptor (VDR) protein in the kidney of neonatal mice, but weight gains of maternal mice were decreased by coumestrol administration. Coumestrol administration increased the messenger RNA (mRNA) expressions of epithelial Ca channels 1 (ECaC1) and VDR in the kidney of neonatal mice, and also increased the mRNA expressions of ECaC2 in the kidney and small intestine of male neonatal mice. The mRNA expressions of ECaC1, ECaC2, calbindin‐D_9k (CaBP‐9k) and estrogen receptor (ER)α in the kidney and VDR in the small intestine of male neonatal mice were higher than those of female mice. Thus, coumestrol administration to maternal mice during pregnancy and lactation may affect renal Ca metabolism in neonatal mice, especially male neonatal mice via maternal milk.     

Studies of calcium-binding protein in cattle

We have developed and validated a double-antibody radioimmunoassay for quantifying bovine calcium-binding protein (CaBP). Cross-reactivity between the antiserum and microgram quantities of thyrocalcitonin, calmodulin, gastrin, cholecystokinin, vasoactive intestinal polypeptide, serum albumin and concentrated extract of bovine pituitary gland was insignificant. Slight cross-reactivity (6%) of the antiserum with parathyroid hormone was demonstrated. Assay sensitivity was .25 ng/ml and intraassay and interassay coefficients of variation ranged from 4 to 11% and 10 to 24%, respectively. The CaBP immunoreactivity was not affected by endogenous Ca concentrations. Plasma and serum concentrations of immunoreactive CaBP were similar. The CaBP concentrations were unaffected when coagulated and anticoagulated blood samples were stored at 4 or 22 C for up to 72 h and when serum was stored at -20, 4 or 22 C for 8 d. Serum CaBP concentrations in cattle were not affected by gonadal steroids, but may have been influenced by age. Treatment with 500 mg of vitamin D3, but not 50 mg of dihydrotachysterol, significantly increased serum Ca and CaBP concentrations in Holstein heifers after a lag period of 7 to 10 d. Serum Ca and CaBP concentrations began to increase in serum at approximately the same time and both exhibited parallel responses to treatment with vitamin D3. Serum Ca concentrations were positively correlated (r = .81) with CaBP concentrations and this relationship was described by the equation, Y = 6.85 + 1.01X - .03X2. Serum Ca and CaBP concentrations were still elevated in heifers 75 d after initial treatment with vitamin D3. The radioimmunoassay we describe provides an opportunity to investigate the role of CaBP in Ca homeostasis during growth, pregnancy, lactation, parturient paresis and other physiological and pathological states in cattle.     

Effects of biological type of beef steers on vitamin D, calcium, and phosphorus status

Feedlot steers (n = 36) from three biological types (Bos indicus, Bos taurus-Continental, and Bos taurus-English) were used to determine the Ca, P, and vitamin D3 status of feedlot cattle. The USDA yield and quality grade traits were measured at slaughter, and the concentrations of vitamin D3 (VITD) and the metabolites 25-hydroxyvitamin D3 (25-OH D) and 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D) were determined in LM, liver, kidney, and plasma. Plasma and muscle Ca and P concentrations also were determined. Biological type of cattle affected a number of carcass traits. Carcasses from Bos taurus-English cattle had more marbling, resulting in higher quality grades (P < 0.05). Carcasses from Bos taurus-Continental cattle had lower calculated yield grades (P < 0.05) than did carcasses from cattle in the other biological types. In general, differences in carcass traits resulting from biological type were consistent with other reports. Plasma and LM Ca and P concentrations were not affected (P = 0.06) by biological type of cattle, indicating that Ca and P homeostasis is a conserved trait across the different types of cattle. Plasma VITD and 25-OH D concentrations were not affected (P = 0.41) by biological type, whereas plasma 1,25-(OH)2 D concentration was lower (P < 0.05) in Bos taurus-English cattle than in Bos taurus-Continental and Bos indicus cattle. Liver VITD and 25-OH D were not affected by biological type (P = 0.76), but liver 1,25-(OH)2 D concentration was greater (P < 0.05) in Bos indicus cattle than in Bos taurus-Continental cattle. Kidney vitamin D metabolite concentrations were not affected by biological type of cattle (P = 0.21). Muscle VITD concentration was greater (P < 0.05) in Bos taurus-English cattle than in the other two biological types, and muscle 25-OH D concentrations were greater (P < 0.05) in Bos taurus-English cattle than in Bos indicus cattle. Muscle 1,25-(OH)2 D concentration was less (P < 0.05) in the Bos taurus-Continental cattle than in the other two biological types. Cooking eliminated vitamin D metabolite differences among the biological types. Our results suggest that Bos indicus cattle had greater 1,25-(OH)2 D (the biologically active form) in tissues, and greater 1,25-(OH)2 D plasma concentrations than Bos taurus cattle. Thus, the need for VITD supplementation and optimal levels of Ca and P in feedlot diets might differ between Bos indicus and Bos taurus cattle.     

磷对奶牛红细胞膜Na^＋,K^＋—ATPase活性影响的观察

观察了土壤、牧草低磷情况下，可影响奶牛血清磷含量。血清磷含量可影响红细胞膜结构与功能。血清磷含量减少会导致Ｎａ＾＋，Ｋ＾＋－ＡＴＰａｓｅ活性降低，进而影响Ｃａ＾２＋－ＡＴＰａｓｅ活性和Ｍｇ＾２＋－ＡＴＰａｓｅ活性。     

Influence of feeding and UVB exposition on the absorption mechanisms of calcium in the gastrointestinal tract of veiled chameleons (Chamaeleo calyptratus)

The purpose of this study was to investigate the influence of feeding and UVB exposition on the occurrence and distribution patterns of vitamin D receptors (VDR) and calbindin D28k (Cb‐D28k) in the gastrointestinal tract of veiled chameleons. Thus, 56 veiled chameleon hatchlings were divided into six treatment groups: UV (with UVB exposure); No (no supplements, no UVB exposure); CaAUV (with calcium (Ca), vitamin A supplementation, UVB exposure); CaA (with Ca, vitamin A supplementation); CaADUV (with Ca, vitamin A, vitamin D supplementation, UVB exposure); and CaAD (with Ca, vitamin A, vitamin D supplementation). Animals were reared under the suspected conditions for 6 months on locust‐based diets. Tissue samples of stomach, duodenum, ileum and colon were taken, and semi‐quantitative immunohistochemical methods (IHC) were performed to detect Cb‐D28k and VDR. VDR immunoreactions were higher in the luminal epithelium of the duodenum than in that of the ileum. VDR immunoreactions in the luminal epithelium were higher at the base of the villi of the duodenum as compared to the tip. Cb‐D28k immunoreactions were mainly observed in the luminal epithelium of the duodenum. The two groups treated with all dietary supplements (CaADUV, CaAD) exhibited a higher Cb‐D28k immunoreaction as those with no supplements and UVB exposure only. No immunoreaction for both proteins could be detected in the stomach. This study suggests that the duodenum plays an important role in the active transcellular absorption of Ca in veiled chameleons as shown by the immunohistochemical detection of VDR and Cb‐D28k. Expression of Cb‐D28k, in particular, appears to be regulated by dietary supplementation of vitamin D and vitamin A. VDRs, however, tended to be upregulated when animals were not supplemented with Ca, vitamin D and vitamin A. This may be due to the decreased Ca concentrations which caused vitamin D activation in the skin without any supplementation, but UVB exposure.     

Effects of bisphenol A administration to pregnant mice on serum Ca and intestinal Ca absorption

Bisphenol A (BPA) is a xenoestrogen commonly used in food storage plastics. The present study was conducted to clarify the effects of BPA administration to pregnant mice on serum calcium (Ca) and Ca metabolism of the gut and kidney. From 6.5 to 16.5 days post coitus (dpc), pregnant mice were administered at 2 mg or 20 mg/kg body weight/day of BPA. Serum Ca was decreased in mice treated with 20 mg BPA at 17.5 dpc, but no remarkable differences were detected in the alkaline phosphatase activity and vitamin D receptor protein expression in the duodenum and jejunum. The messenger RNA (mRNA) expressions of calcium binding protein (CaBP‐9k) and active vitamin D synthesis enzyme (CYP27B1) in the kidney were increased in mice treated with 20 mg BPA. The mRNA expressions of occludin and junction adherence molecular A (JAM‐A) in the duodenum and ileum, which regulate paracellular transport, were increased in mice treated with 20 mg BPA. However, the administration of 2 mg BPA had no effect on serum Ca and mRNA expressions of relative genes in Ca metabolism. These results imply that BPA administration at 20 mg/kg body weight/day during pregnancy decreases serum Ca in pre‐delivery mice, which may be partly due to decreased paracellular Ca absorption.     

Prevention of goitre in new-born lambs from kale-fed ewes

Milk urea concentrations in dairy cattle. There has been increased use of milk urea concentration as an indicator of dietary protein intake and protein metabolism in dairy cattle over recent years. The value of milk urea content data in predicting dietary composition, particularly for pasture-fed cattle, has not been well described.

Protein metabolism and urea synthesis. Many factors influence the degradation of dietary proteins in the rumen, post-ruminal protein metabolism and urea synthesis in cattle. Strong positive correlations between nitrogenous fertiliser use and the crude protein content of pastures were identified by use of meta-analysis. Similar strong positive correlations were noted between dietary protein intake, rumen ammonia and plasma urea concentrations. The costs of urea synthesis include energy losses, and importantly, the loss of endogenous amino acids, which are deaminated in the synthesis of urea.

Milk urea as an indictor of protein metabolism. Urea concentrations in blood, plasma and milk are strongly correlated. Milk is an adequate indicator of blood and plasma urea content, but non-nutritional factors may significantly influence milk urea concentrations. Recommendations for dietary protein management based on milk urea concentrations must be undertaken with care.     

The use of vitamin D3 and its metabolites to improve beef tenderness

Three experiments were conducted to determine whether feeding 25-hydroxyvitamin D3 (25-OH D3) or 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D3) improves the tenderness of longissimus dorsi (LD), semimembranosus (SM), and infraspinatus (IF) muscles similar to supplemental vitamin D3 without leaving residual vitamin D3 and its metabolites in muscle. In the first two experiments, 24 crossbred steers were used to determine the effects of different oral amounts of 1,25-(OH)2 D3 (Exp. 1; n = 12) and 25-OH D3 (Exp. 2; n = 12) on plasma Ca2+ concentrations. In the third experiment, crossbred steers were allotted randomly to one of four treatments: 1) control placebo (n = 7); 2) 5 x 10(6) IU of vitamin D3/d (n = 9) for 9 d and harvested 2 d after last treatment; 3) single, 125-mg dose of 25-OH D3 (n = 8) 4 d before harvest; or 4) single, 500-microg dose of 1,25-(OH)2 D3 (n = 9) 3 d before harvest. The LD and SM steaks from each animal were aged for 8, 14, or 21 d, whereas steaks from the IF were aged for 14 or 21 d. All steaks were analyzed for tenderness by Warner-Bratzler shear force and for troponin-T degradation by Western blot analysis. Supplementing steers with vitamin D3 increased (P < 0.01) the concentration of vitamin D3 and 25-OH D3 in all muscles sampled. Feeding steers 25-OH D3 increased (P < 0.05) the concentration of 25-OH D3 in meat, but to an amount less than half that of cattle treated with vitamin D3. Supplemental 1,25-(OH)2 D3 did not affect (P < 0.10) shear force values; however, there was a trend (P < 0.10) for supplemental vitamin D3 and 25-OH D3 to produce LD steaks with lower shear values after 8 and 14 d of aging, and lower (P < 0.10) shear force values for the SM aged for 21 d. Analysis of Western blots indicated that LD steaks from cattle supplemented with vitamin D3 and 25-OH D3 had greater (P < 0.05) troponin-T degradation. Antemortem supplementation of 25-OH D3 seems to increase postmortem proteolysis and tenderness in the LD and SM without depositing large concentrations of residual vitamin D3 and its metabolite 25-OH D3.     

Hormonal regulation of calcium homeostasis in two breeds of dogs during growth at different rates

Growing giant-breed dogs are more susceptible to developing skeletal disorders than small-breed dogs when raised on diets with deficient or excessive Ca content. Differential hormonal regulation of Ca homeostasis in dogs with different growth rates was investigated in Great Danes (GD, n = 9) and Miniature Poodles (MP, n = 8). All animals were raised on the same balanced diet and under identical conditions. Calciotropic and growth-regulating hormones were measured. Production and clearance of 1,25-dihydroxycholecalciferol (1,25[OH]2D3) were investigated with the aid of [3H]-1,25(OH)2D3 and renal messenger RNA abundance of 1 alpha-hydroxylase and 24-hydroxylase. Intestinal, renal, and skeletal Ca handling were evaluated with the aid of 45Ca balance studies. Skeletal development was evaluated by radiology and histomorphometry. Great Danes had greater (P < 0.001) growth rates than MP, as indicated by the 17-fold greater body weight gain, by increased longitudinal growth reflected in the increased (P < 0.05) gain in length of the radius and ulna, and by increased (P < 0.001) growth plate thickness. These findings were accompanied in GD by greater (P < 0.05) plasma GH and IGF-I concentrations. Effects were observed for vitamin D3 metabolism, such as greater (P < 0.01) plasma 1,25(OH)2D3 concentrations due to decreased (P < 0.01) clearance rather than increased production of 1,25(OH)2D3, and decreased (P < 0.01) plasma 24,25-dihydroxycholecalciferol (24,25[OH]2D3) concentrations likely due to competitive inhibition of the production of 24,25(OH)2D3. These findings were accompanied in both breeds by a limited hormonal regulation of Ca and P absorption at the intestinal level, and in GD by increased (P < 0.05) renal reabsorption of inorganic P (Pi) compared with MP, resulting in greater (P < 0.01) Pi retention and greater (P < 0.01) plasma Pi concentrations. Bone turnover, resorption, and formation were greater (P < 0.01) in GD than in MP. In addition, GD had more irregular (P < 0.01) growth plates than MP, accompanied by disorders of endochondral ossification. It is suggested that in GD, increased calcitonin levels and/or a relative deficiency in 24,25(OH)2D3 at the growth-plate level may both be responsible for the retarded maturation of chondrocytes, resulting in retained cartilage cones and osteochondrosis, and this may be a pathophysiological factor for the increased susceptibility of large breed dogs to developing skeletal disorders.     

Indications and implications for testing of milk urea in dairy cattle: a quantitative review. Part 1. Dietary protein sources and metabolism

MILK UREA CONCENTRATIONS IN DAIRY CATTLE: There has been increased use of milk urea concentration as an indicator of dietary protein intake and protein metabolism in dairy cattle over recent years. The value of milk urea content data in predicting dietary composition, particularly for pasture-fed cattle, has not been well described. PROTEIN METABOLISM AND UREA SYNTHESIS: Many factors influence the degradation of dietary proteins in the rumen, post-ruminal protein metabolism and urea synthesis in cattle. Strong positive correlations between nitrogenous fertiliser use and the crude protein content of pastures were identified by use of meta-analysis. Similar strong positive correlations were noted between dietary protein intake, rumen ammonia and plasma urea concentrations. The costs of urea synthesis include energy losses, and importantly, the loss of endogenous amino acids, which are deaminated in the synthesis of urea. MILK UREA AS AN INDICATOR OF PROTEIN METABOLISM: Urea concentrations in blood, plasma and milk are strongly correlated. Milk is an adequate indicator of blood and plasma urea content, but non-nutritional factors may significantly influence milk urea concentrations. Recommendations for dietary protein management based on milk urea concentrations must be undertaken with care.     

Vitamin D receptor amounts across different segments of the gastrointestinal tract in Brown Swiss and Holstein Frisean cows of different age

During different stages of lactation, different requirements of calcium have to be met depending on the milk amount. Vitamin D receptors (VDR) regulate calcium homeostasis by increasing the entry of Ca into blood from bone stores and dietary sources. The purpose of this study was to investigate if age and breed of cows influence VDR amounts across different segments of the gastrointestinal tract. Thirty-six cows were used (18 Brown Swiss, 18 Holstein Friesan, both > 5.5 years or < 4.5 years). Tissue specimens of the intestines were collected from the cows. Formaldehyde-fixed and microwave-treated paraffin sections were used for VDR immunohistochemistry employing a biotinylated monoclonal rat antibody and streptavidin peroxidase technique. The results showed that nuclei and cytoplasm of enterocytes stained positively for VDRs. Strongest immunoreactions were observed in intermediate and basal glandular cells. No significant differences were observed between the different groups. Vitamin D receptors immunoreactivities were prominent in duodenal mucosa, lower in jejunum and in colon, decreased further in ileum and were lowest in caecum. Decreases in number of positively marked cells and staining intensities resulted in reduced immunoreactions. The results of this study indicate that VDR are highly expressed at the site of maximal intestinal calcium absorption. No significant influence of age and breed was observed. The animals used were not in a negative Ca balance. The cows were all in the stage of late or mid lactation. During these periods, the Ca requirements are low and the diets are high in Ca concentration; and the animals are adapted to these circumstances. Passive absorption in adult animals seems to dominate when Ca intake is adequate or high. The active absorption may play a considerably more significant role during the peripartal period, when Ca homeostatic mechanisms are challenged because of tremendous Ca demand at the initiation of lactation.     

Calcitonin mrna expression and plasma calciotropic hormones in fetal lambs

The object of this study was to measure basal levels of calciotropic hormones during gestation in the ovine fetus. Thyroid glands and plasma samples were collected from 49 Ile de France x (Limousine x Romanov) fetuses (range, 80 to 145 d). Total thyroidal RNA was extracted for calcitonin (CT) mRNA analysis. Plasma immunoreactive concentrations of CT, parathyroid hormone (PTH), parathyroid hormone-related peptide (PTHrP), and 1,25-dihydroxy-vitamin D [1,25(OH)ZD] were assayed. Plasma Ca and P concentrations were measured by atomic absorption spectrophotometry and colorimetry, respectively.The ratio of thyroidal weight/body weight decreased twice between Days 80 and 145 of gestation. Simultaneously, the CT mRNA rate increased but did not significantly vary after 89 d of gestation. Plasma CT concentration increased between 80 and 89 d and then did not significantly vary until Day 145. We could not detect PTH in ovine fetuses plasma; PTHrP concentration did not vary significantly between Days 80 and 145. The concentration of 1,25-(OH)₂D decreased in plasma essentially between Days 110 and 145. In conclusion, between Days 80 and 145 of gestation, CT, PTHrP, and 1,25-(OH)₂D concentrations can be easily measured in plasma from ovine fetuses, but plasma PTH was undetectable. In basal conditions, no relationship could be demonstrated between any hormonal concentrations in the fetal plasma and that of Ca or P.     

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.     

Calcium metabolism of pregnant rats fed a vitamin D-depleted diet

The effects of vitamin D (VD) deficiency on calcium (Ca) metabolism during pregnancy were evaluated in rats fed VD-repleted (VD-repleted rats) and VD-depleted (VD-depleted rats) diets. In the VD-depleted rats, the plasma concentrations of 1,25-dihydroxyvitamin D and Ca decreased severely, whereas the parathyroid hormone concentrations increased. The Ca contents of the feces of the VD-depleted rats were higher than those of the VD-repleted rats. The fetal Ca contents of the VD-repleted and VD-depleted rats increased continuously, but that of the VD-depleted rats was lower. These data reveal that VD deficiency during pregnancy induces severe hypocalcemia due to reduced intestinal absorption of Ca and elevated fetal demand for Ca.