Dose-response effects of an aqueous formulation of calcitriol and evaluation of renal function in nonpregnant cows

Dose responses of plasma calcitriol, calcium (Ca), bone metabolic markers and glomerular filtration rate (GFR) were evaluated in four nonpregnant Holstein cows treated subcutaneously with an aqueous formulation of calcitriol at four doses in a 4 × 4 Latin-square design. Calcitriol, Ca, and markers of bone metabolism were analyzed in plasma samples. GFR was measured in predose and day 5 samples. Plasma calcitriol and Ca concentrations increased dose-dependently. The calcitriol dose was positively correlated with the area under the concentration-time curve of plasma calcitriol. Bone formation markers tended to increase from day 3 onward for all doses. No significant changes in GFR were noted. Thus, exogenous calcitriol administered between 0.0625 and 0.5 μg/kg body weight elicited dose-dependent increases in both plasma calcitriol and Ca and elevated bone formation markers without affecting renal function in nonpregnant cows.     

Effects of 1,25-dihydroxyvitamin D3 on calcium and phosphorus homeostasis in sheep fed diets either adequate or restricted in calcium content

It was the aim of the present study to collect basic data on calcium (Ca) and phosphorus (P) homoeostasis in sheep. Two series of experiments were carried out to investigate the effects of 1,25-dihydroxyvitammin D₃ (calcitriol) in supraphysiological dosage in combination with varying alimentary Ca supply. In the first series, blood samples were collected over 72 h to determine the concentrations of total Ca (Ca), ionized Ca (Ca²⁺), inorganic phosphate (P_i), and the bone resorption marker CrossLaps (CL). In the second series, measurements were carried out over 12 h. In addition, urine samples were collected to calculate the fractional excretions (FE) of Ca and P_i. Changes in plasma macromineral concentrations (P < 0.01) as well as in CL (P < 0.001) and endogenous calcitriol (P < 0.05) were observed in the alimentary Ca-restricted animals, indicating that the reduction of daily Ca intake challenged the animals’ macromineral homeostatic mechanisms. However, the Ca-restricted diet had an effect on neither FE of Ca nor on FE of P_i. The treatment resulted in peak serum calcitriol concentrations between 1,900 and 2,500 pg/mL, and supraphysiological concentrations were maintained for the next 48 h. Irrespective of dietary Ca, calcitriol had hypercalcemic and hyperphosphatemic effects. An increase in CL was revealed only in the Ca-restricted, calcitriol-treated sheep (P < 0.01), reflecting a remarkable enhancement of Ca mobilization from the bone by calcitriol exclusively in this group. From these data, it can be concluded that the sheep can be a suitable animal model for studying catabolic effects of Ca deficiency and calcitriol on bone metabolism.     

Association between changes in plasma calcium concentration and plasma tartrate-resistant acid phosphatase activity in periparturient cows

Eight periparturient Holstein Friesian cows were examined for plasma tartrate-resistant acid phosphatase (TRAP) activity to assess the degree of bone metabolic activity and to evaluate the association between the change in calcium (Ca) concentration and bone metabolism during the periparturient period. Milk fever occurred in 1 of 8 cows just after parturition. Plasma TRAP activities did not markedly change in 5 of 8 cows during the experimental period. The changing rate of Ca between preparturition and just after parturition was under -20% in 3 of 8 cows, and low TRAP activities were observed in 2 of these 3 cows. This study suggests that cows with a low TRAP activity are at risk of developing milk fever in comparison to cows with high TRAP activity. Temporary increases of parathyroid hormone were observed in 7 cows, but not in the cow with milk fever.     

Serum activities of tartrate-resistant acid phosphatase and bone specific alkaline phosphatase as indices of bone metabolism in the cow

The correlation between the serum hydroxyproline concentration and serum activity levels of TRAP and BALP was examined in 41 cows. The correlated coefficient (r) was 0.6391 for TRAP and 0.3147 for BALP, respectively. Judging from the significant correlation to the serum hydroxyproline concentration, serum TRAP activity is an index for bone metabolism in cows. Serum TRAP activity was therefore measured in 205 healthy cows (2-9 years old) in order to observe the changes in bone resorption with aging and milk production. TRAP levels differed slightly between group A (< or =4 yrs) and B (5 yrs< or =) at the same stage of lactation. The activity levels rose slightly at the height of lactation stage and during the dry stage.     

The course of some bone remodelling plasma metabolites in healthy horses and in horses offered a calcium-deficient diet

An inquiry was carried out to assess the concentrations of plasma metabolites related to bone remodelling in 21 saddle horses of Warmblood breed aged 4-26 years, five draught horses of Ardennes breed aged 4-10 years, and 10 Ardennes foals aged 9-11 months. They were fed according to normal feeding practice in Belgium. The changes in some bone remodelling plasma metabolite concentrations were studied when an unbalanced diet was offered and later corrected for four Warmblood horses. Bone formation was evaluated by bone alkaline phosphatase (BALP), total alkaline phosphatase (TALP) and osteocalcin (bone gla-protein, OC). Bone resorption was assessed by hydroxyproline (HYP). Total calcium, ionized calcium, phosphorus (P) and 25-hydroxyvitamin D3 [25-(OH)D] concentrations were more or less constant. The comparison of four bone remodelling factors between the Ardennes and Warmblood horses showed higher concentrations in the Ardennes breed. Bone marker concentrations decreased according to age. The correction of the unbalanced Ca : P diet induced inconsistent effects at plasma level. The interpretation of the different bone parameters appeared to be difficult if not associated with other parameters such as a complete anamnesis and clinical examination of the animal in addition to dietary evaluation.     

Changes in the plasma levels of several bone markers in newborn calves during the first two days of life

The fluctuations in the plasma levels of several bone markers were investigated in newborn calves. Experiment 1 monitored the postnatal changes in the plasma levels of tartrate-resistant acid phosphatase isoform 5b (TRAP5b), total alkaline phosphatase (t-ALP) and bone-specific alkaline phosphatase (BAP) in four calves. These markers increased significantly from 9−20 hr after the first colostrum-suckling compared with the values immediately after birth. Experiment 2 evaluated changes in the plasma TRAP5b, t-ALP, BAP and type I collagen cross-linked N-telopeptide (NTx) levels within 2 days post-birth in five calves with successful passive immunization via colostrum (non-deficient group) and five others with poor colostrum intake (deficient group). The non-deficient group had significantly higher plasma levels of the four parameters around 12 hr of life compared with the deficient group. The results suggest that the increase in plasma bone markers in calves in the first day of life is related to the colostrum intake.     

Plasma fluctuation in estradiol-17β and bone resorption markers around parturition in dairy cows

Blood samples were obtained sequentially from 10 dairy cows around the time of parturition to assess plasma fluctuations in estradiol-17β (E₂) levels in association with those of several bone resorption markers. Plasma E₂ concentration increased sharply a few days prepartum and decreased quickly after parturition. In terms of bone resorption markers, the plasma level of tartrate-resistant acid phosphatase isoform 5b (TRAP5b) rose significantly, commencing 1 week prepartum, and was maintained at this level to a few days postpartum. The plasma concentration of carboxyterminal collagen cross-links of type-I collagen (CTx) increased significantly after parturition. These observations suggest that osteoclast-mediated bone resorption was activated after parturition when plasma E₂ concentrations decreased.     

Changes of serum alkaline phosphatase activity in dry and lactational cows

Serum alkaline phosphatase (ALP) activities were measured during dry and lactational periods to investigate the influence of lactation on serum ALP activity in cows. Higher levels of serum ALP activity were seen in lactational periods than in dry periods. The serum activities of bone-specific ALP (BALP), liver ALP (LALP), tartrate resistant acid phosphatase (TRAP) and aspartate aminotransferase also increased in lactational periods. ALP activities in the bone extract and in whey were decreased at similar rates by the addition of lectin. Moreover, since the ALP band in whey was observed to have the same migration in polyacrylamide gel (PAG) disk electrophoresis as that of the bone extract, analysis of ALP isoenzymes by lectin affinity or PAG disk electrophoresis could not distinguish ALP originating from the mammary gland from that of bone. In this study, it was clear that the increased level of serum ALP activity was due to increases of BALP and LALP in lactational periods. However, the extent of the influence of ALP originating from the mammary glands on serum ALP activity was unknown. Judging from changes of BALP and TRAP activities in the serum and the correlation between the both, it was guessed that ALP originating from the mammary glands influenced serum ALP activity.     

Effects of calcium intake and parity on plasma minerals and bone turnover around parturition

Eight pregnant heifers (primiparous cows) and seven pregnant cows in their second, third and forth pregnancies (multiparous cows) were assigned to two groups and fed either a low calcium (Ca) diet (Ca, 0.46%) or a high Ca diet (Ca, 0.86%) ad libitum from 3 weeks before the expected calving date to 3 days after parturition. All cows were examined for a change in dry matter intake (DMI), plasma minerals and bone turnover around parturition. The dietary Ca level did not affect the DMI in both primiparous and multiparous cows. The DMI of primiparous cows was significantly lower than that of multiparous cows (P < 0.05) in both the low and high dietary Ca groups. The dietary Ca level did not affect the concentrations of plasma Ca, phosphorus, magnesium and parathyroid hormone throughout the experimental period. Plasma phosphorus in primiparous cows was significantly higher (P < 0.05) than that of multiparous cows around parturition. Plasma Ca and magnesium tended to be higher (P < 0.10) in primiparous cows. The dietary Ca level did not affect the plasma osteocalcin (OC) level measured as bone formation or the urinary deoxypyridinoline (DPD) excretion measured as bone resorption before parturition in both primiparous and multiparous cows. After parturition, the plasma OC level was lower than it was before parturition in multiparous cows fed a low Ca diet, and in primiparous cows. There were no significant differences in urinary DPD excretion between each group before and after parturition. Both the plasma OC level and urinary DPD excretion of primiparous cows were significantly higher than those of multiparous cows in both the low and high dietary Ca groups.     

过瘤胃维生素D对围产期奶牛低血钙症的影响

为了阐明日粮添加过瘤胃维生素D对围产期奶牛低钙血症的调控机制,在某集约化奶牛场选取40头奶牛,随机分为4组,对照组(C)饲喂基础日粮,试验Ⅰ、Ⅱ、Ⅲ组分别于产前10d至分娩后10d在日粮中每天添加1.5、3.0、6.0g/头过瘤胃维生素D。对4组试验奶牛围产期(分别于产前10、7、3、0d和产后3、7、10d)血清维生素D3(1,25-(OH)2D3)、钙、羟脯氨酸、甲状旁腺素和降钙素水平进行检测。结果表明,围产期奶牛血钙水平分娩时达最低,产后逐渐上升,且产前7d至产后10d对照组奶牛血钙浓度显著低于试验Ⅰ组(P<0.05);分娩当天试验Ⅰ组奶牛血浆中1,25-(OH)2D3、羟脯氨酸和降钙素水平显著高于对照组(P<0.05),甲状旁腺素水平显著低于对照组(P<0.05)。提示,日粮中每天添加1.5g/头过瘤胃维生素D能调节甲状旁腺素和降钙素等激素的分泌,促进肠道对钙的吸收,动员骨钙,从而防止奶牛产后低钙血的发生,且围产期奶牛低血钙症与机体血钙稳态调节作用密切关联。     

Oral 1,25-dihydroxyvitamin D3 in prevention of milk fever

The effect of daily oral doses of 1,25-dihydroxy vitamin D₃ (1,25-(OH)₂D₃) on plasma mineral concentrations and incidence of milk fever was tested in 39 aged cows. Three dose levels of 1,25-(OH)₂D₃ (0, 100 and 200 µg/d) were compared in cows supplemented with 100 g Ca/d (250 g CaCO₃) and in cows receiving no Ca supplement. 1,25-(OH)₂D₃ treatments were given from day 5 before expected calving (day –5) until the day after calving (Calving = day 0).Expected minima in Ca concentrations in plasma were seen on day 1 in placebo treated cows, while Ca concentrations in 1,25-(OH)₂D₃ treated cows were normal or increased in the period from start of treatment until day 3–5. In the parturient period (day –1 to +2) 1 of 21 treated cows developed milk fever, as compared to 4 of 18 untreated. A pronounced hypocalcaemia developed, however, in the 1,25-(OH)₂D₃ treated cows from day 3 to 5 onwards, culminating day 8–11 with 7 cases of milk fever. Ca supplements reduced the development of hypocalcaemia in the 1,25-(OH)₂D₃ treated cows, but could not completely prevent the occurrence of milk fever.     

The change of dry matter intake, milk yield and bone turnover in primiparous cows compared to multiparous cows during early lactation

Data from six primiparous and nine multiparous Holstein cows were used to clarify the difference of Ca and P mobilization between primiparous and multiparous cows during early lactation. The dry matter intake (DMI) of primiparous cows was lower (P < 0.01) than those of multiparous cows. Milk yield was lower in primiparous cows at 7, 14, 21 (P < 0.01), and 28 days (P < 0.05) after parturition. There was no significant difference in milk Ca and P concentrations between primiparous and multiparous cows. There were no significant differences in plasma Ca and P concentrations between primiparous and multiparous cows. The plasma P level at 7 days postpartum in primiparous cows was lower (P < 0.05) than 28 days postpartum. The concentration of plasma osteocalcin (OC) measured as bone formation marker of primiparous cows was significantly higher than multiparous cows (P < 0.01) at 21 and 28 days postpartum. The urinary deoxypyridinoline (DPD) as bone resorption marker of primiparous cows tended to be higher (P < 0.10) than multiparous cows at 21 days after parturition and decreased to the same level as that of multiparae toward the peak lactation. These results show that Ca and P mobilization of primiparous cows are more active than multiparous cows.     

Effects of anionic salts supplementation on blood pH and mineral status, energy metabolism, reproduction and production in transition dairy cows

The objectives of this study were to determine the effects of a pre-partum diet with lower than recommended (DCAD = −82 mEq/kg of dietary DM) amounts of anionic salts on metabolism, health, reproductive performance and milk production in dairy cows. Sixty Holstein multiparous cows were enrolled 21 days prior to expected calving date. The animals were randomly assigned to receive one of two rations: 30 cows received anionic ration [−82 mEq (NA + K − Cl − S)/kg of DM] for 21 d to parturition and the other group (n = 30) were fed a usual dry period ration (+192 mEq/kg of DM). Serum samples obtained at days −21, +3 and +21 relative to calving were analyzed for β-hydroxybutyrate (BHBA), non-esterified fatty acids (NEFA), glucose, calcium (Ca), inorganic phosphorus, magnesium, chloride, sodium, potassium, cholesterol, urea, creatinine, total protein, albumin, and aspartate aminotransferase (AST). Urine pH declined from 8.4 at 21 d before calving (pre-treatment) to 6.2 at day 7 pre-partum in the treatment group. Repeated-measure mixed model analysis indicated that the concentrations of Ca were significantly increased and creatinine, and AST were significantly decreased by lowering DCAD. The concentrations of BHBA, NEFA and glucose were not affected by treatment. The incidence of milk fever and culling were 5 and 11 times higher in the control group in comparison with the treatment group, respectively. The intervals from calving to first breeding and to pregnancy were not influenced by treatment. There was no group effect on average daily milk yield or fat percentage.     

Effects of cortisol secreted via a 12-h infusion of adrenocorticotropic hormone on mineral homeostasis and bone metabolism in ovariectomized cows

To evaluate the effects of endogenously secreted cortisol on mineral homeostasis and bone metabolism in cows, 4 ovariectomized Holstein cows were infused for 12 h with either an adrenocorticotropic hormone (ACTH) solution (0.5 mg/2 L isotonic NaCl solution per cow) or isotonic NaCl solution in a 2 × 2 crossover design. ACTH infusion stimulated cortisol secretion and increased plasma cortisol concentrations for 18 h (P < 0.001), leading to an elevated plasma glucose concentration until 36 h (P < 0.001). Plasma calcium and magnesium concentrations in ACTH-infused cows fluctuated within normal ranges, whereas hypophosphatemia was observed unequivocally. The biochemical bone resorption markers tartrate-resistant acid phosphatase 5b and hydroxyproline decreased following ACTH infusion (P < 0.001 and P = 0.003, respectively). Similarly, the bone formation marker, bone-specific alkaline phosphatase, decreased continuously until 72 h after the ACTH infusion (P < 0.001). These results demonstrate that increased secretion of cortisol via a 12-h ACTH infusion disrupted homeostasis of inorganic phosphate and suppressed bone metabolism in ovariectomized cows without involving gonadal steroid hormones.     

The course of selected bone resorption marker concentrations in response to short-term hypocalcemia experimentally induced with disodium EDTA infusions in dairy cows

The collagen metabolites hydroxyproline (HYP), deoxypyridinoline (DPD), and the carboxyterminal telopeptide of type I collagen (ICTP) are suitable markers for bone resorption in humans and several animal species. The purpose of this study was to describe the course of bone resorption markers during short-term hypocalcemia induced with disodium ethylenediaminetetraacetic acid (Na2EDTA) and to investigate whether bone resorption is increased in dairy cows under these conditions. EDTA infusions have been used as a model for periparturient paresis in dairy cows and to estimate the calcium mobilization rate from body reserves in ruminants. In this study, hypocalcemia was induced by means of a 5% Na2EDTA infusion (0.55 mg/kg/min Na2EDTA for 5 h = total dose of 100.6 g). Two experiments were conducted: (1) Six 4-11 years-old Brown Swiss cows were infused intravenously with EDTA for 5 h. Blood and urine samples were taken repeatedly from 1 day before until 10 days after infusion. (2) Towards the end of the lactation, the experiment was repeated with the same animals after a 14-day-period of feeding a low calcium diet (26 g/animal per day). The EDTA-infusion induced hypocalcemia and hypophosphatemia. The HYP-, DPD- and ICTP-concentration remained mainly unaffected during both infusions. Only DPD showed an increase during infusion and HYP an increase 2 days after the infusion. In conclusion, the EDTA infusion had little effect on the concentrations of the measured bone markers, which may be due to the fact that the serum calcium pool was refilled by increased absorption of Ca via the gastrointestinal tract. From these results, it can be concluded that bone resorption was not influenced by EDTA infusion.     

Intravaginal administration of lactic acid bacteria modulated the incidence of purulent vaginal discharges,plasma haptoglobin concentrations,and milk production in dairy cows

This investigation studied the effects of intravaginal administration of a mixture of lactic acid bacteria (LAB) on the incidence of purulent vaginal discharges (PVD), plasma haptoglobin concentrations, and milk production in dairy cows. A total of 82 pregnant primiparous and multiparous Holstein dairy cows were used in this study. Half of the cows received intravaginally 1 mL of LAB at 10¹⁰–10¹² cfu/mL and the other half 1 mL of reconstituted skim milk (i.e., carrier) (controls). Administration of LAB was conducted once per wk during 2 and 1 wk before the expected day of calving and at 1, 2, 3, and 4 wk postpartum. Data demonstrated that intravaginal administration of LAB decreased the occurrence of PVD at 3 wk postpartum (P < 0.05). Concentrations of plasma haptoglobin, an acute phase protein often associated with uterine infections, was lower in cows treated with the LAB mixture at 2 wk (P < 0.001) and 3 wk (P < 0.05) postpartum. Treatment with LAB did not improve overall pregnancy rate, but the treated multiparous cows produced more milk than their control counterparts (P < 0.05), whereas no difference was observed in primiparous cows regarding milk yield (P > 0.05). Overall, this is the first study demonstrating that intravaginal LAB administration lowers the incidence of PVD and enhances milk production in dairy cows. Further research is warranted to evaluate the effects of LAB on reproductive performance in a larger cohort of cows.     

Biomarkers for the activation of calcium metabolism in dairy cows: elevation of tartrate-resistant acid phosphatase activity by lowering dietary cation-anion difference is associated with the prevention of milk fever

In our previous study, it was demonstrated that the administration of anion salts, which slightly lower the dietary cation-anion difference (DCAD), in the prepartum period is safe and effective for preventing milk fever in multiparous cows. In the present study, several biomarkers, which might show activation of Ca metabolism, were analyzed using stored samples in the previous study to investigate the mechanism of the preventive effect on milk fever by lowering DCAD. Changes in bone-specific alkaline phosphatase activity, osteocalcin and insulin-like growth factor I concentrations in serum were almost the same among the three groups of multiparous cows with or without the oral administration of anion salts, while the levels of these serum biomarkers in the group of primiparous cows (heifer group) were much higher compared with those in the three multiparous groups throughout the experimental period. Urinary deoxypyridinoline excretion was not a useful biomarker for dairy cows because it hardly changed during the peripartum period in all groups. However, serum tartrate-resistant acid phosphatase (TRAP) activity, which is known as a biomarker of osteoclast activity, was well associated with the administration of anion salts lowering DCAD because among the three multiparous groups, only the group of multiparous cows fed the anion salts (anion group) showed an increased level, which rose to the level in the heifer group, and was markedly higher than those in the other control groups of multiparous cows. The increased activity of serum TRAP in the anion group suggested that Ca in the plasma pool was mobilized smoothly from bone-bound Ca via mature osteoclasts at parturition, which might be due to prior activation under mild acidosis induced by slightly lowering DCAD. Therefore, TRAP was the best biomarker to monitor the activation of Ca metabolism in dairy cows fed anion salts.     

Diurnal variation in concentrations of various markers of bone metabolism in dogs.

OBJECTIVE: To evaluate diurnal variation in concentrations of selected markers of bone metabolism in dogs. ANIMALS: Ten 3- to 4-year-old ovariectomized Beagles. PROCEDURE: Blood and urine samples were obtained in the morning before dogs were fed (8 AM) and then at 2-hour intervals for 24 hours. This procedure was repeated 2 weeks later. Concentrations of osteocalcin (OC) and carboxy terminal telopeptide of type-I collagen (ICTP) were measured in serum, using a radioimmunoassay; concentrations of hydroxyproline (HYP), pyridinoline (PYD), and deoxypyridinoline (DPD) were analyzed in urine. Hydroxyproline concentration was measured by means of a colorimetric test, whereas PYD and DPD concentrations were quantified by use of high-performance liquid chromatography. RESULTS: In both parts of the study, HYP concentrations increased significantly, compared with values before feeding, until 8 hours after feeding; HYP concentrations then returned to prefeeding values. Concentrations of DPD and PYD decreased from before feeding until 2 PM and then increased until 8 PM. The ICTP concentrations slowly decreased until 4 PM but returned to prefeeding values thereafter. In both parts of the study, concentrations of OC decreased during the day and then increased to reach values similar to those obtained before feeding. CONCLUSIONS: Changes in the concentrations of bone markers were detected throughout the day in the dogs of this study. Increase in HYP concentration most likely was related to feeding. As documented for bone resorption and formation in other species, circadian rhythms were evident for concentrations of DPD, PYD, and OC. Investigators should consider the time of sample collection when measuring these markers.     

The effect of high ambient temperature on Ca,P and Mg balance and bone turnover in high‐yielding dairy cows

We investigated the effect of heat stress on Ca, P and Mg balance and bone turnover in lactating cows. In a 2 × 2 crossover design, four multiparous lactating Holstein cows were kept in a chamber and subjected to a constant moderate (18°C) ambient temperature (MT) or high (28°C) ambient temperature (HT). The cows were fed total mixed ration (Ca, 0.7%; P, 0.4%; Mg, 0.2%) ad libitum. The milk yield under HT (35.4 kg/day) tended to be lower (P < 0.10) than that under MT (43.2 kg/day). The concentrations of milk P (P < 0.05) and Mg (P < 0.01) were significantly lower under HT than MT. The Ca, P and Mg intake (P < 0.10); Ca (P < 0.10), P, and Mg (P < 0.05) secretion into milk; and Ca (P < 0.05), P (P < 0.01), and Mg (P < 0.05) absorption in the intestine were lower under HT than MT. The plasma osteocalcin, a marker of bone turnover, was significantly lower (P < 0.05) under HT than MT. Heat stress did not affect plasma C‐telopeptide of collagen type I, a bone resorption marker, and plasma parathyroid hormone concentration.