Effects of maternal and offspring treatment with two dietary sources of vitamin D on the mineral homeostasis, bone metabolism and locomotion of offspring fed protein- and phosphorus-reduced diets.

The present study aims to compare the effects of maternal and offspring treatment with 25-hydroxycholecalciferol (25-OHD3) and vitamin D3 on vitamin D status, mineral homoeostasis, bone metabolism and locomotion in the offspring. Either vitamin D3 (50 µg/kg diet) or 25-OHD3 (50 µg/kg diet) was supplemented to the gestation and lactation diets of 49 multiparous sows and/or to the diets of their growing offspring. Treatment of the sows did not affect plasma concentrations of 25-OHD3 of the offspring. Pigs fed 25-OHD3 had higher plasma concentrations of 25-OHD3 than pigs that received vitamin D3 during rearing and fattening. However, neither plasma concentrations of calcium, phosphate and bone markers during the observation period nor bone ash and bone mineral density at slaughter were clearly affected by the treatment. Maternal and offspring treatment with 25-OHD3 instead of vitamin D3 resulted in a slight reduction in the prevalence of leg swelling. In addition, more pigs walked with even steps and normal stride length. Further studies are needed to test whether the slight effects observed in the present experiment are reproducible and of relevance for animal health and welfare. In that case, the underlying mechanisms should be revealed in order to take advantage of potentially beneficial influences especially under certain feeding regimes.

Dietary supplementation of 25-hydroxycholecalciferol as an alternative to cholecalciferol in swine diets: A review.

25-hydroxycholecalciferol (25-OHD3 ) formed via hepatic hydroxylation from vitamin D, cholecalciferol, represents the precursor of the biologically active vitamin D hormone, 1,25-dihydroxyvitamin D. Due to a higher absorption rate and the omission of one hydroxylation, dietary supplementation of 25-OHD3 instead of vitamin D3 is considered to be more efficient as plasma concentrations of 25-OHD3 are increased more pronounced. The present review summarises studies investigating potential beneficial effects on mineral homeostasis, bone metabolism, health status and performance in sows, piglets and fattening pigs. Results are inconsistent. While most studies could not demonstrate any or only a slight impact of partial or total replacement of vitamin D3 by 25-OHD3 , some experiments indicated that 25-OHD3 might alter physiological processes when animals are challenged, for example, by a restricted mineral supply.

Comparative Study of the Effects of Two Dietary Sources of Vitamin D on the Bone Metabolism, Welfare and Birth Progress of Sows Fed Protein- and Phosphorus-Reduced Diets.

To investigate the influence of two dietary sources of vitamin D on the vitamin D status, bone metabolism, welfare and birth progress of gestating and lactating sows, forty-nine multiparous sows were randomly assigned to one of two diets: "CON" (n = 25; 50 µg vitamin D3/kg feed) and "HYD" (n = 24; 50 µg 25-hydroxycholecalciferol/kg feed). The basal diets were protein- and phosphorus-reduced. The trial started on day 3 ante insemination of the sows and ended with weaning of the piglets on day 28 postpartum. Dietary supplementation of 25-hydroxycholecalciferol resulted in improved maternal vitamin D status (p < 0.001), fewer gait changes (p < 0.01) and longer standing time after feeding (day 5 ante partum; p < 0.05) compared to vitamin D3. However, the bone markers CrossLaps and osteocalcin were not affected. Overall, the present results suggest that sows fed 25-hydroxycholecalciferol instead of vitamin D3 showed improved locomotion and stance strength. However, this outcome is probably not related to altered bone metabolism. The underlying mechanisms must be investigated in further studies.

Egg yolk colour in organic production as affected by feeding - Consequences for farmers and consumers.

In organic table egg production, saponified extracts of carotenoids are not allowed to intensify egg yolk colour. Therefore, we investigated the suitability of organically produced marigold flower meal (Tagetes erectus, TE) and spinach (Spinacia oleracea, SO) as carotenoid sources (mixture of 25% TE and 75% SO) to reach values of 9-10 'Roche Yolk Colour Fan' units (RYCF units). Feeding a completely unsupplemented control diet resulted in a yolk colour of 4.7 RYCF units, a total supply of 11.1 g/kg, 14.6 g/kg and 17.5 g/kg of TE and SO as stand-alone carotenoid sources induced a significant increase to 8.0, 8.2 and 8.9 RYCF units. Under 'winter-feeding conditions', i.e. minimal carotenoid supply in the diet, 17.5 g/kg TE and SO resulted in 9.3 ± 0.7 RYCF units. It can be concluded that supplementing TE and SO is suitable to improve the yolk colour in organically produced table eggs.

Calcinosis in Alpaca Crias (Vicugna pacos) Due to Vitamin D Intoxication-Clinical, Laboratory and Pathological Findings with a Focus on Kidney Function.

Alpacas kept in Central Europe are often deficient in vitamin D3, which is supplemented orally or by injection by the owners or veterinarians. Vitamin D3 can be specified in two different units (IU and µg), which differ by a factor of 40. By mixing up these units, an overdosage can be induced. In this study, three alpaca crias were examined after vitamin D3 intoxication, with particular reference to kidney function. All three animals developed non-specific clinical alterations 1-2 weeks after a vitamin D3 overdose of approximately 40 times. Plasma of the animals revealed several alterations. The main findings were severe azotemia, hypercalcemia and hyperphosphatemia, 15 days after treatment. Kidney function analysis (endogenous creatinine clearance) in two of the crias revealed severe glomerular damage. All crias died despite intensive treatment within 23 days after vitamin D3 treatment. Necropsy revealed calcification in different organs, mainly the kidneys, lungs and liver. Since nine other crias in the same group were treated with comparable doses of vitamin D3 and no clinical signs were observed in these animals, it is concluded that individual animals show different levels of sensitivity to vitamin D3.

An investigation on the relevance of prolactin, insulin-like growth factor-1 and 25-hydroxyvitamin D3 (25-OHD3 ) in canine benign prostatic hyperplasia in a predisposed breed model.

Serum concentrations of prolactin (PRL), insulin-like growth factor-1 (IGF-1) and 25 hydroxyvitamin D3 (25-OHD3 ) were analysed to investigate their possible involvement in the pathogenesis of benign prostatic hyperplasia (BPH). For this, dogs of the Rhodesian Ridgeback (RR) breed were used because of a verified breed disposition for the development of BPH. Labrador Retrievers (LR) served as controls. The prostate gland status was characterised by the prostate gland volume, clinical signs of BPH (haemospermia and sonographic findings) and the plasma concentration of canine prostate-specific arginine esterase (CPSE). Breed specificity in the RR was expressed by a correlation of PRL with breed (p < 0.05). Similar relationships existed in the dogs with normal CPSE (CPSEn) with respect to the IGF-1 concentrations (LR: p < 0.05). The latter were negatively correlated with prostatic volume and age (both p < 0.05). Concentrations of 25-OHD3 were tendentially (p = 0.18) lower in the RR with increased CPSE (CPSEi) compared with the CPSEn LR and RR showing clinical signs of BPH. A negative correlation between serum 25-OHD3 and age (p < 0.05) existed in the CPSEi RR. Proof of 25-OHD3 in prostatic secretion proved to be a breed specific feature in the RR (p < 0.0001). For all RR dogs showing clinical signs of BPH, a close to significant (p = 0.06) positive correlation with prostate gland volume was found. The results of the present study reveal no clear hints towards the significance of PRL and IGF-1 in the pathogenesis of canine BPH. In the RR breed there were indications of a causal relationship with age-dependent changes in the vitamin D metabolism. The data suggest the possibility of preventing or treating canine BPH by administering vitamin D or substances involved in the intraprostatic vitamin D metabolism.

Low potassium diets with different levels of calcium in comparison with different anionic diets fed to prepartum dairy cows: Effects on sorting behaviour, total tract digestibility, energy metabolism, oxidative status and hormonal response.

This study investigated the effects of low potassium diets with different levels of Ca compared to two diets low in dietary cation-anion difference (DCAD) fed prepartum as a strategy to prevent hypocalcemia on sorting behaviour, total tract digestibility, oxidative status and energy and protein metabolism of transition cows. Forty-eight pregnant dairy cows were assigned to 4 treatment groups: Low Ca, low K (LCLK), High Ca, low K (HCLK), Supplementation with anionic mineral mixture (AMS) supplementation with SoyChlor (CAS). After parturition, all animals were fed a standard postpartum diet. Data were collected until 21 DIM. Prepartum urinary pH was significantly reduced by the low DCAD diets, while postpartum Ca homeostasis was affected by the HCLK ration. Feeding AMS induced sorting against particles <1.18 mm in favour of particles >19 mm prepartum. In contrast, cows fed CAS showed an increase in selective consumption of fine particles and sorted against longer particles similar to the HCLK and LCLK groups. Postpartum sorting activity was not affected by the dietary treatments. After calving, apparent digestibility of NDF was significantly reduced in the HCLK group. Prepartum, we observed effects on serum concentrations of non-esterified fatty acids were higher and insulin sensitivity was lower in the AMS group. Blood urea nitrogen (BUN) was decreased in cows fed the CAS ration. Postpartum, we found serum protein to be decreased with the low DCAD diets while BUN was decreased in the CAS group. The low DCAD rations increased prepartum serum malondialdehyde concentrations, while postpartum total antioxidant capacity was lower in the HCLK and the AMS group. From these data, we conclude that AMS decreased prepartum intake due to compromised palatability. Intermediate protein metabolism was affected by the low DCAD diets, while parameters of oxidative stress were probably affected by acid-base balance and Ca homeostasis.

Dietary phosphorus restriction affects bone metabolism, vitamin D metabolism and rumen fermentation traits in sheep.

Homeostasis of calcium (Ca) and phosphate (Pi ) is maintained by a concerted interplay of absorption and reabsorption via the gastrointestinal tract and the kidney and by storage and mobilization from the bone regulated mainly by parathyroid hormone (PTH), 1,25-dihydroxycholecalciferol and calcitonin. The present study aimed at characterizing the effects of dietary P restriction on bone, vitamin D metabolism and rumen fermentation traits reflecting the endogenous P cycle maintaining the ruminal P supply for microbial metabolism. The experiments were done in eleven female, non-pregnant, non-lactating four- to nine-year-old Black Headed Mutton sheep allotted to two feeding groups: "P-restricted" (0.11% P/kg DM and 0.88% Ca/kg DM) and "Control" (0.38% P/kg DM and 0.88% Ca/kg DM). Dietary P restriction did not lead to hypophosphataemia, probably due to a compensation by bone mobilization, demonstrated by increased serum concentrations of a resorption marker and altered gene expression in bone tissue. In addition, the RNA expression of fibroblast growth factor 23, a bone-derived factor involved in the regulation of vitamin D metabolism, was significantly reduced with dietary P restriction. Furthermore, several genes related to vitamin D metabolism and plasma concentrations of 1,25-(OH)2 D were associated with serum concentrations of phosphate (Pi ). In the parotid gland, the expression of the Pi transporter NaPi2b was negatively associated with serum Pi and positively with parathyroid PTH expression. Although Pi concentrations in saliva and the gastrointestinal tract were significantly reduced, we found no adverse effects of the P-restricted ration on the production of short chain fatty acids, but slight differences in the production of butyrate as well as its relationship to rumen Pi and ammonia concentrations that might indicate an impact on ruminal fermentation.

Serum parameters related to mineral homeostasis and energy metabolism in ewes kept on different dietary magnesium supply during the transition period.

The present study investigated the effects of dietary magnesium supplementation on mineral status and intermediary metabolism of ewes during the transition period (TP). For this purpose, 19 pregnant ewes (2nd and 3rd lactation) were divided into a control group (Con, n = 9) kept on a daily magnesium intake of approximately 3.0 g ante-partum (a.p.) and 5.0 g post-partum (p.p.), and a magnesium group (Mg, n = 10, 4.2 g a.p., 7.7 g p.p.). Blood was collected at day (d) 30 a.p., d 14 a.p., d 1 p.p., d 14 p.p. and d 30 p.p., respectively. Serum levels of total magnesium were not affected by the treatment. In Con animals, serum levels of total calcium and phosphate were lower at d 1 p.p. in comparison to d 30 a.p. (P ≤ 0.05). Only in Con ewes, serum glucose levels showed significant fluctuations throughout the entire observation period (P ˂ 0.05) and were correlated negatively with those of beta-hydroxybutyrate (r2: 0.70; P < 0.01, r2: 0.76; P < 0.01) and positively with insulin (r2: 0.46; P < 0.05, r2: 0.59; P < 0.05) on d 30 and 14 a.p., respectively. At d 30 p.p., serum insulin levels were lower in the Mg group compared to the Con group (P < 0.05). As serum concentrations of calcium, phosphate and glucose seemed more stable throughout the TP, magnesium supplementation might be an approach to promote metabolic health in ewes.

Blood leukocyte composition and function in periparturient ewes kept on different dietary magnesium supply.

BACKGROUND: Transition period (TP) is characterised by physiological and metabolic changes contributing to immunodysregulation. Since knowledge about this period in sheep is scarce, we analysed changes in selected immune variables during the TP in ewes and whether dietary magnesium (Mg) supplementation could modulate these immune variables. Pregnant ewes (2nd and 3rd lactation) were divided into a control group (CONT, n = 9) and a Mg group (MAG, n = 10) supplemented with Mg oxide resulting in a daily Mg intake of approximately 0.30 and 0.38% (MAG) of dry matter during ante- (a.p.) and post-partum (p.p.) periods, respectively. Blood samples were collected between days (d) 30 a.p. and d 30 p.p.. Whole blood neutrophil phagocytic activity, monocyte subset (classical cM, intermediate intM, non-classical ncM) composition and the proliferative capacity of lymphocytes were determined flow cytometrically. At d 14 a.p., all ewes were vaccinated against Mycobacterium avium subsp. paratuberculosis (MAP). RESULTS: Both groups showed a sharp increase in the total leukocyte counts (TLC) and neutrophil counts (P < 0.0001), at d 1 p.p., while, monocytes and their subpopulations displayed the highest values at d 30 p.p. (P ≤ 0.05). At d 1 p.p. the neutrophil phagocytic activity was higher (P < 0.05) in MAG ewes. Throughout the TP, the proliferative response of CD4+ cells was significantly higher in the MAG group (P < 0.05). Ewes in both groups responded with an increase in the TLC, neutrophil numbers (P ≤ 0.05) and ncM (P < 0.001) 24 h post vaccination, whereas monocytes and cM dropped in numbers (P ≤ 0.05). Numbers of intM only increased in MAG ewes (P < 0.05), whereas lymphocyte numbers decreased (P < 0.01). Mg supplementation did not affect the significant increase in MAP-specific antibodies at d 7 and 21 post vaccination. Total Mg and Ca serum levels did not show any differences between the two groups. CONCLUSION: Whereas TP-associated fluctuations in blood leukocyte numbers are not influenced by Mg supplementation, neutrophil phagocytic activity, the proliferative capacity of CD4+ cells and the cellular response within 24 h after a vaccination are subject to modulation.

Calcinosis in a roe deer fawn (Capreolus capreolus) in northern Germany.

BACKGROUND: Calcinosis has been reported for a broad range of different animals. Causes for calcinosis include metabolic disorders due to kidney failure, intoxication with calcinogenic plants, or iatrogenic overdose of vitamin D. Especially young animals seem to be very susceptible to developing calcinosis. Currently, however, there is a lack of information on calcinosis in wildlife. CASE PRESENTATION: The following case report describes a roe deer fawn admitted to a clinic due to general weakness and myiasis. Plasma levels for creatinine, urea and phosphate were highly elevated, whereas the total calcium level was decreased. Necropsy revealed calcinosis due to calcification in many organs. The reason for calcinosis in this particular case might be kidney failure. Plasma samples from other hunted roe deer fawns also showed high phosphate levels. CONCLUSIONS: Roe deer fawns might be susceptible to calcinosis due to high plasma phosphate, which could be a result of kidney failure or different feed. Further research into calcium and phosphate homeostasis in roe deer is necessary.

Bone Health or Performance? Adaptation Response of Genetically Divergent Chicken Layer Lines to a Nutritive Calcium Depletion.

In modern laying hybrids, calcium (Ca) homeostasis is immensely challenged by daily eggshell calcification. However, excessive mobilization of Ca from bones may lead to osteoporosis, which then manifests in a high incidence of poor bone quality. The aim of this study was to characterize the hens' adaptation response to an alternating dietary Ca restriction. The animal model consisted of four purebred layer lines, differing in laying performance (high vs. moderately performing lines) and phylogenetic origin (white- vs. brown-egg lines). According to the resource allocation theory, hens selected for high egg production were assumed to show a different response pattern to cope with this nutritive challenge compared to moderately performing lines. Data collected included egg number, egg quality traits, body weight and bone characteristics. The Ca depletion led to a temporary drop in egg production and shell quality and a loss of bone stability due to Ca mobilization. The white-egg lines response was more pronounced, whereas the brown-egg lines were less sensitive towards reduced Ca supply. Our study shows that the hens' responsiveness to coping with a nutritive Ca depletion is not ultimately linked to genetic selection for increased egg production but rather to phylogenetic origin.

Impact of a Ration Negative in Dietary Cation-Anion Difference and Varying Calcium Supply Fed before Calving on Colostrum Quality of the Dams and Health Status and Growth Performance of the Calves.

This study investigated the effect of diets negative in dietary cation-anion difference (DCAD) or restricted in Ca fed prepartum to dairy cows for three weeks on colostrum yield and composition, and the health and growth performance of their calves. Thirty-six pregnant non-lactating Holstein-Friesian cows were randomly assigned to three isoenergetic diets: (1) low Ca: 0.24% Ca, DCAD: +86 mEq/kg; (2) high Ca: 1.23% Ca, DCAD: +95 mEq/kg; and (3) low DCAD: 1.28% Ca, DCAD: -115 mEq/kg (all dry matter (DM) basis). While colostrum quality was not affected, low Ca supply prepartum tended to increase the colostrum yield compared to high Ca (low Ca = 8.81 vs. high Ca = 5.39 kg). However, calves from cows fed low DCAD showed higher serum concentrations of K, lower body weight (BW), starter feed intake and average daily weight gain before weaning compared to low Ca and high Ca calves (53.12 vs. 57.68 and 57.32 kg) but BW was similar postweaning (d 70). In addition, calves from dams fed low DCAD were more likely to develop diarrhea and had increased number of days with abnormal fecal scores. Consequently, calves from low DCAD dams had to be treated more frequently.

Symposium review: Transition cow calcium homeostasis-Health effects of hypocalcemia and strategies for prevention.

The effects of subclinical hypocalcemia have been explored in numerous observational and mechanistic studies in recent years. Besides obvious, well-known effects on muscle contractility, the role of Ca with respect to immune function and intermediary metabolism explains the contribution of subclinical hypocalcemia to the development of several diseases observed in early lactation and underlines its importance in high-performing dairy cows. The present review aims at integrating recent scientific progress, such as discoveries about the role of the mammary gland in regulating bone mobilization, into generally accepted aspects of the endocrine control of Ca homeostasis. We will discuss Ca transport mechanisms through absorption, resorption, secretion, and mobilization, as well as the physiological regulation of Ca through parathyroid hormone, 1,25-dihydroxyvitamin D, fibroblast growth factor 23, and serotonin, in addition to dietary mineral requirements. To improve hypocalcemia prevention strategies, our knowledge of the physiological mechanisms necessary to maintain normocalcemia and their endogenous regulation should be combined with data derived from herd-level studies. Using such studies, we will discuss prepartum nutritional strategies aimed at reducing the incidence of subclinical hypocalcemia, as well as options for postpartum Ca supplementation and their effects on early-lactation health and production. Especially in respect to approaches that might interfere with endogenous adaptation processes, such as supplementation with vitamin D metabolites or large doses of Ca, a thorough understanding of the underlying mechanisms that might induce unwanted hypocalcemia rebound effects will be crucial to ameliorate our future management of transition cows. Continued efforts by researchers to understand the interaction of Ca homeostasis with prevention strategies is necessary to optimize cow health and support copious milk production.

Dietary protein and calcium modulate parathyroid vitamin D receptor expression in young ruminants.

For economic reasons and in order to minimize nitrogen excretion and thus pollution, the crude protein content in the diet of livestock animals should be as low as possible without negatively affecting the animals´ health and performance. As ruminants can efficiently use dietary protein because of the ruminohepatic circulation of urea, they are considered to cope more easily with such a feeding regime than monogastric animals. However, despite unaltered daily weight gain, massive changes in mineral homeostasis and vitamin D metabolism were observed with dietary protein reduction (N-) in young, growing goats. Serum concentrations of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) were decreased with a low N intake, even if calcium (Ca) was also restricted (Ca-). Interestingly, concentrations of cyclic adenosine monophosphate (cAMP) measured as an indirect assessment for the parathyroid hormone (PTH) activity were not affected by low protein. Therefore, it was hypothesized that the sensitivity of the parathyroid gland is modulated during these dietary interventions. Four groups of male German colored goats received a control (N+/Ca+), a reduced protein (N-/Ca+), a reduced Ca (N+/Ca-) or a reduced protein and Ca (N-/Ca-) diet. After six weeks we determined the expression of PTH, PTH receptor, Ca sensing receptor (CASR), vitamin D receptor (VDR), retinoid X receptor (RXRα), Klotho, fibroblast growth factor receptor 1c-splicing form, and the sodium-dependent Pi transporter (PiT1) in the parathyroid glands. Concentrations of cAMP were not affected, while those of Ca and 1,25-(OH)2D3were diminished and that of 25-hydroxyvitamin D3 was increased with N- feeding. The expression patterns of the described target genes were not altered. In contrast, animals fed the Ca- rations showed enhanced serum 1,25-(OH)2D3 and cAMP levels with no changes in blood Ca concentrations demonstrating an efficient adaptation. The mRNA expression of expression of VDR and CASR in the parathyroid gland was significantly diminished and RXRα, PTHR and PiT1 expression was elevated. Instead of the assumed desensitization of the parathyroid gland with N-, our results indicate elevated responsiveness to decreased blood Ca with feeding Ca-.

In vivo measurement of strontium absorption from the rumen of dairy cows as an index of calcium absorption capacity.

Absorption of dietary calcium from the rumen is a quantitatively important process in calcium homeostasis of ruminants. In 3 separate experiments in dairy cows, we applied a technique developed in sheep to measure the rate of strontium (Sr) absorption from the rumen as an indicator of calcium absorption capacity. Absorption from the rumen after an oral dose of SrCl2 resulted in a maximum plasma concentration of Sr after 1 h, whereas absorption from the small intestine after injection of SrCl2 into the abomasum through a cannula occurred more slowly. The second experiment demonstrated that the calcium absorption capacity index of the rumen was significantly greater in 21 lactating Friesian cows (230 ± 66, mean ± SEM) than in 6 mature, nonlactating, nonpregnant heifers (101 ± 21, mean ± SEM). In a third experiment, we compared clinically normal cows at the onset of lactation with those that developed parturient paresis. In cows that developed severe hypocalcemia, plasma concentrations of 1,25(OH)2D were significantly elevated (144 ± 60 pg/mL vs. 90 ± 54 pg/mL; means ± SEM) and their rumen calcium absorption index was significantly decreased compared with that of clinically normal cows. Evidence suggested that mobilization of calcium from bone as lactation commenced was significantly depressed in paretic cows compared with those that did not show clinical signs of hypocalcemia. Moreover, ruminal stasis suppressed the absorption of calcium from the rumen. We conclude that measurement of Sr concentration in blood plasma after an oral dose of SrCl2 into the rumen can be used as an index of rumen calcium absorption capacity under different states of calcium homeostasis.

A reduced protein diet modulates enzymes of vitamin D and cholesterol metabolism in young ruminants.

Besides other adverse effects, a low protein diet has been shown to modulate cholesterol and vitamin D metabolism in monogastric species like rats and humans. As ruminants can increase the efficiency of the rumino-hepatic circulation of urea, it is assumed that goats should be able to compensate for a low dietary protein intake better. After a dietary protein restriction (9% vs. 20%) for six weeks, plasma concentrations of urea, albumin, 1,25-dihydroxyvitamin D3 and calcium were decreased, while plasma 25-hydroxyvitamin D3 (25-OHD3), and total cholesterol were significantly increased in young goats. Because this was not accompanied by any decrease in expression of CYP24A1 mRNA, we investigated mRNA expression of additional enzymes with known 24- and/or 25-hydroxylase activities (CYP2R1, CYP2J2, CYP3 A24, CYP27A1), receptors involved in their regulation (VDR, PXR, RXRα) and vitamin D binding protein (VDBP). CYP2R1expression was stimulated with the low dietary protein intake, negatively correlated with plasma urea and positively associated with serum 25-OHD3. The greater plasma concentrations of total cholesterol could be explained with the reduction of CYP2J2 and CYP27A1 expression. None of the receptors investigated were affected by the dietary protein restriction but mRNA expression of VDBP was slightly reduced. Taken together our results show that dietary protein restriction has an impact on vitamin D and cholesterol metabolism in ruminants, too. Therefore, further investigations are needed before dietary interventions aiming at diminishing nitrogen excretion can be implemented.

Gastrointestinal transport of calcium and glucose in lactating ewes.

During lactation, mineral and nutrient requirements increase dramatically, particularly those for Ca and glucose. In contrast to monogastric species, in ruminants, it is rather unclear to which extend this physiological change due to increased demand for milk production is accompanied by functional adaptations of the gastrointestinal tract (GIT). Therefore, we investigated potential modulations of Ca and glucose transport mechanisms in the GIT of lactating and dried-off sheep. Ussing-chamber technique was applied to determine the ruminal and jejunal Ca flux rates. In the jejunum, electrophysiological properties in response to glucose were recorded. Jejunal brush-border membrane vesicles (BBMV) served to characterize glucose uptake via sodium-linked glucose transporter 1 (SGLT1), and RNA and protein expression levels of Ca and glucose transporting systems were determined. Ruminal Ca flux rate data showed a trend for higher absorption in lactating sheep. In the jejunum, small Ca absorption could only be observed in lactating ewes. From the results, it may be assumed that lactating ewes compensate for the Ca loss by increasing bone mobilization rather than by increasing supply through absorption from the GIT Presence of SGLT1 in the jejunum of both groups was shown by RNA and protein identification, but glucose uptake into BBMV could only be detected in lactating sheep. This, however, could not be attributed to electrogenic glucose absorption in lactating sheep under Ussing-chamber conditions, providing evidence that changes in jejunal glucose uptake may include additional factors, that is, posttranslational modifications such as phosphorylation.

Expression of Tight Junction Proteins and Cadherin 17 in the Small Intestine of Young Goats Offered a Reduced N and/or Ca Diet.

Diets fed to ruminants should contain nitrogen (N) as low as possible to reduce feed costs and environmental pollution. Though possessing effective N-recycling mechanisms to maintain the N supply for rumen microbial protein synthesis and hence protein supply for the host, an N reduction caused substantial changes in calcium (Ca) and phosphate homeostasis in young goats including decreased intestinal transepithelial Ca absorption as reported for monogastric species. In contrast to the transcellular component of transepithelial Ca transport, the paracellular route has not been investigated in young goats. Therefore, the aim of the present study was to characterise the effects of dietary N and/or Ca reduction on paracellular transport mechanisms in young goats. Electrophysiological properties of intestinal epithelia were investigated by Ussing chamber experiments. The expression of tight junction (TJ) and adherens junction (AJ) proteins in intestinal epithelia were examined on mRNA level by qPCR and on protein level by western blot analysis. Dietary N reduction led to a segment specific increase in tissue conductances in the proximal jejunum which might be linked to concomitantly decreased expression of cadherin 17 mRNA. Expression of occludin (OCLN) and zonula occludens protein 1 was increased in mid jejunal epithelia of N reduced fed goats on mRNA and partly on protein level. Reduced dietary Ca supply resulted in a segment specific increase in claudin 2 and claudin 12 expression and decreased the expression of OCLN which might have been mediated at least in part by calcitriol. These data show that dietary N as well as Ca reduction affected expression of TJ and AJ proteins in a segment specific manner in young goats and may thus be involved in modulation of paracellular Ca permeability.

Dietary nitrogen and calcium modulate bone metabolism in young goats.

Ruminants, possessing the rumino-hepatic circulation, are thought to cope easily with reduced dietary nitrogen (N) supply which is of economic and environmental interest to diminish N output. Nevertheless, feeding an N reduced diet to young goats resulted in a decrease in calcitriol and calcium (Ca) plasma concentrations. Although a dietary Ca reduction alone stimulated calcitriol synthesis and plasma Ca concentrations were restored, in combination with a reduced N supply this stimulating effect was abolished. Based on the important role bone tissue plays in maintaining Ca homeostasis, aim of the present study was to determine effects of an N reduced diet with or without a concomitant Ca reduction on bone metabolism in young goats. A dietary N reduction alone resulted in a significant rise in plasma concentrations of bone resorption marker C-terminal telopeptide of type I collagen (CTX) and bone formation marker osteocalcin (OC), while reduced intake of Ca as well as the combination of both dietary interventions increased bone markers only slightly. Bone mineral content and bone mineral density of metatarsi were decreased by reduced N intake, while Ca and phosphorus (P) content of dried bones remained unaffected. In contrast, a dietary Ca reduction alone led to decreased Ca and P content of dried bones. From these data it can be concluded that a dietary N reduction alone or in combination with a reduced dietary Ca supply modulated bone metabolism in young goats.