Evaluation of ear skin temperature as a cow-side test to predict postpartum calcium status in dairy cows.

Subclinical hypocalcemia is considered a gateway disease that increases susceptibility to other metabolic and infectious diseases in transition dairy cows. In the absence of a cow-side test, however, it is difficult to identify hypocalcemic cows. The objective of this study was to evaluate ear skin temperature as a diagnostic predictor of serum calcium concentration. We conducted a cross-sectional study on 7 commercial dairy farms, involving 251 cows 0 to 48h after calving. Skin temperature of the ears (STEar) was scored manually by palpating both ears. An infrared thermometer was used to measure ear temperature, skin temperature on the coxal tuber (STCox), and ambient temperature. Rectal temperature was measured using a digital thermometer. A blood sample was drawn to determine serum calcium concentration. Hypocalcemia was defined as serum calcium below 2.0mmol/L, irrespective of clinical symptoms. Serum calcium concentration <2.0mmol/L in connection with clinical symptoms was defined as clinical milk fever; serum calcium concentration <2.0mmol/L without clinical symptoms was defined as subclinical hypocalcemia. Multivariate analysis using the GENLINMIXED procedure and receiver operating characteristic analysis were performed to evaluate whether serum calcium concentration could be predicted using ear temperature and other temperature estimates. The prevalence of hypocalcemia was 3.3, 27.3, 32.8, and 69.6% for cows in first, second, third, and fourth or greater lactation, respectively. None of the cows in first and second lactation had clinical milk fever. The prevalence of clinical milk fever was 6.0 and 20.3% for cows in their third and fourth or greater lactation, respectively. A decrease in ear temperature of 0.39°C [95% confidence interval (CI): 0.25-0.54] was associated with a decrease of 0.1mmol/L in serum calcium concentration. Ambient temperature, however, was a major confounder for ear temperature. With an increase in ambient temperature of 1°C, STEar rose by 0.78°C (95% CI: 0.67-0.90). Hypothermia was more pronounced in clinical milk fever (median 21.8°C; interquartile range 14.7-27.0°C) compared with subclinical hypocalcemia (median 27.6°C, interquartile range 22.1-30.8°C). All temperature estimates had only accurate test characteristics based on their area under the curve for prediction of subclinical hypocalcemia (area under the curve for STEar, STCox, and rectal temperature were 0.641, 0.668, and 0.606, respectively) when cows with clinical milk fever were excluded. Although ear temperature has been associated with serum calcium concentration, ear temperature cannot be recommended for diagnosis of subclinical hypocalcemia.

Microarray-based gene expression profiling of peripheral blood mononuclear cells in dairy cows with experimental hypocalcemia and milk fever.

Although a molecular diagnostic assay using clinically accessible tissue, such as blood, would facilitate evaluation of disease conditions in humans and animals, little information exists on microarray-based gene expression profiling of circulating leukocytes from clinically hypocalcemic cows. Therefore, peripheral blood mononuclear cells from dairy cows with experimentally induced hypocalcemia or spontaneous milk fever were subjected to oligo-microarray analysis to identify specific biomarker genes. In experimental hypocalcemia induced by a 4-h infusion of 10% disodium EDTA (n=4), 32 genes were significantly up- or downregulated compared with control treatment (4-h infusion of 11% calcium EDTA; n=4). In cows with milk fever (n=8), 98 genes were expressed differentially (either up- or downregulated) compared with healthy parturient cows (n=5). From these data, the following 5 genes were selected as being strongly related to both experimental hypocalcemia and milk fever: protein kinase (cAMP-dependent, catalytic) inhibitor ß (PKIB); DNA-damage-inducible transcript 4 (DDIT4); period homolog 1 (PER1); NUAK family, SNF1-like kinase, 1 (NUAK1); and expressed sequence tag (BI537947). Another gene (neuroendocrine secretory protein 55, NESP55) was also determined to be specific for milk fever, independently of hypocalcemia. The mRNA expression of these 6 genes in milk fever cases was verified by quantitative real-time reverse-transcription PCR and was significantly different compared with their expression in healthy parturient cows. In the present study, the selected genes appeared to be candidate biomarkers of milk fever because the continuous interactions between blood cells and the entire body suggest that subtle intracellular changes occur in association with disease. However, before any genomic biomarkers are incorporated into clinical evaluation of the disease, the effect of hypocalcemia on the mRNA expression of these genes in the tissues that regulate calcium homeostasis in dairy cows should be determined.

Diet-induced pseudohypoparathyroidism: A hypocalcemia and milk fever risk factor.

Subclinical hypocalcemia may affect half of all multiparous cows, and clinical hypocalcemia or milk fever affects approximately 5% of dairy cows each year. This disorder of calcium homeostasis can be induced by several dietary factors. Recent studies implicate high dietary potassium and high dietary cation-anion difference (DCAD) with increased risk of milk fever. The hypothesis tested in this study was that high-DCAD diets fed to prepartum cows reduce tissue sensitivity to parathyroid hormone (PTH), inducing a pseudohypoparathyroid state that diminishes calcium homeostatic responses. Multiparous Jersey cows were fed low- or high-DCAD diets in late gestation, creating a compensated metabolic alkalosis in the high-DCAD cows and a compensated metabolic acidosis in the low-DCAD cows. They then received synthetic PTH injections at 3-h intervals for 48 h. Parathyroid hormone is expected to cause an increase in plasma calcium by increasing renal production of 1,25-dihydroxyvitamin D and increasing bone calcium resorption. Plasma calcium concentration increased at a significantly lower rate in cows fed the high-DCAD diet. Cows fed the high-DCAD diet also produced significantly less 1,25-dihydroxyvitamin D in response to the PTH injections than cows fed the low-DCAD diet. Serum concentrations of the bone resorption marker carboxyterminal telopeptide of type I collagen were numerically lower in cows fed the high-DCAD diet but this difference was not statistically significant. These data provide direct evidence that high-DCAD diets reduce tissue sensitivity to PTH. The metabolic alkalosis associated with high-DCAD diets likely induces a state of pseudohypoparathyroidism in some dairy cows at the onset of lactation, resulting in hypocalcemia and milk fever.

Towards sustainable and humane dairy farming: A low-cost electrochemical sensor for on-site diagnosis of milk fever.

Milk fever is a metabolic disorder that predominantly affects dairy animals during the periparturient period and within four weeks of calving. Milk fever is primarily attributed to a decrease in the animal's serum Ca2+ levels. Clinical milk fever occurs when Ca2+ concentration drops below 1.5 mM (6 mg/dL). Without prompt intervention, clinical milk fever leads to noticeable physical symptoms and health complications including coma and fatality. Subclinical milk fever is characterized by Ca2+ levels between 1.5 and 2.12 mM (6-8.48 mg/dL). Approximately 50% of multiparous dairy cows suffer from subclinical milk fever during the transition to lactation. The economic impact of milk fever, both direct and indirect, is substantial, posing challenges for farmers. To address this issue, we developed a low-cost electrochemical sensor that can measure bovine serum calcium levels on-site, providing an opportunity for early detection of subclinical and clinical milk fever and early intervention. This calcium sensor is a scalable solid contact ion sensing platform that incorporates a polymeric calcium-selective membrane and ionic liquid-based reference membrane into laser-induced graphene (LIG) electrodes. Our sensing platform demonstrates a sensitivity close to the theoretical Nernstian value (29.6 mV/dec) with a limit of detection of 15.6 µM and selectivity against the species in bovine serum. Moreover, our sensor can detect Ca2+ in bovine serum with 91% recovery.

Short communication: Circulating serotonin (5-HT) concentrations on day 1 of lactation as a potential predictor of transition-related disorders.

The monoamine serotonin (5-hydroxytryptamine; 5-HT) has been described as a homeostatic regulator of lactation. Recently, our laboratory determined that 5-HT is involved in the regulation of calcium and glucose homeostasis during the transition period in rodents. More specifically, we demonstrate that 5-HT is responsible for calcium mobilization from bone and upregulation of hepatic gluconeogenic enzymes and mammary gland glucose transporters. Our objective was to investigate the correlation between circulating 5-HT concentrations and circulating ionized calcium, parathyroid hormone-related protein (PTHrP), and glucose concentrations on d 1 postpartum. We also investigated the correlation between circulating 5-HT and milk fever and ketosis incidence and severity in multiparous Holstein cows at the onset of lactation. Blood samples were collected from 42 multiparous cows on d 1 of lactation and analyzed for 5-HT, calcium, glucose, and PTHrP. Milk fever (determined subjectively for each cow on d 1 postpartum) and ketosis incidence and severity (scale 1 to 4, determined objectively for each cow during the first 10 d postpartum) were recorded for all animals. Serum 5-HT was positively correlated with serum calcium and with plasma PTHrP (r>0.37). Serum 5-HT was negatively correlated with milk fever incidence and with ketosis severity (most severe ketosis incidence recorded during the first 10 d postpartum; r<-0.33). Serum calcium and plasma glucose concentrations were negatively correlated with milk fever and ketosis severity, respectively (r<-0.39). These data indicate that 5-HT potentially plays a role in the regulation of calcium and glucose homeostasis during the transition period in cattle, which we previously demonstrated in rodents. Increased circulating concentrations of 5-HT might decrease milk fever at the onset of lactation and ketosis severity during the first 10 d postpartum in dairy cows. Understanding this physiological axis could help describe the underlying mechanisms associated with these periparturient metabolic disorders in dairy cows.

Treatment of cows with milk fever using intravenous and oral calcium and phosphorus.

Fifteen cows with milk fever were treated with 500ml of 40 % calcium borogluconate (group A) administered intravenously. Fifteen other cows with milk fever received the same treatment, supplemented with 500ml of 10 % sodium phosphate administered intravenously, and 80g calcium as calcium lactate and 70g inorganic phosphorus as sodium phosphate administered orally in drinking water. The cows were monitored and blood samples collected for 3 days to measure the concentrations of total and ionized calcium, inorganic phosphorus and magnesium and the activity of creatine kinase. The two groups did not differ significantly with respect to the course of the disease. In each group 14 cows were cured. A rapid and significant increase in serum calcium concentration from the hypo- to the hypercalcaemic range occurred in both groups within 10min of the start of treatment, followed by a slow and steady decrease to the hypocalcaemic range. Calcium lactate did not prevent the calcium concentration from returning to the hypocalcaemic range, and the calcium profiles of the two groups did not differ significantly. As expected, treatment had little effect on the concentration of inorganic phosphorus in group A. In group B, treatment caused a rapid increase in the concentration of inorganic phosphorus to a maximum 20min after the start of treatment. This was followed by a slow decrease in the phosphorus concentration to the normophosphataemic range. Our findings confirmed that combined intravenous and oral administration of sodium phosphate in cows with periparturient paresis attributable to hypocalcaemia and hypophosphataemia results in a rapid and sustained increase in serum phosphorus, but not in serum calcium concentration. This modified therapy did not improve the success rate of milk fever treatment and further studies are needed to improve treatment of periparturient paresis.

[The effect of oral calcium lactate in postpartum cows]. / Untersuchungen zur Wirkung von Kalziumlaktat bei Kühen post partum.

The oral administration of calcium lactate for prophylaxis of bovine parturient paresis has been promoted for a number of years. The goal of the present study was to investigate the effect of this treatment on the serum concentrations of calcium, inorganic phosphorus and magnesium in parturient cows. Five fresh calved cows were given a drench containing calcium lactate and 5 control cows received the same drench but without calcium lactate. There were no significant differences in the serum concentrations of total and ionised calcium and magnesium between treated and control cows within 24 hours of treatment. Because the calcium lactate drench did not significantly affect calcium concentrations in the blood of fresh cows, it appears highly questionable whether the administration of calcium lactate decreases the risk of bovine parturient paresis. Based on our results, the oral administration of calcium lactate can not be recommended for prophylaxis of bovine parturient paresis.

Blood markers of osteoclastic differentiation in periparturient dairy cows at different parities, with and without milk fever.

Osteoprotegerin (OPG) inhibits osteoclast (OC) differentiation. TRAP5b (tartrate-resistant acid phosphatase 5b) secreted by OCs reflects the numbers of mature OCs. This study assessed these OC-related markers around parturition in cows of different parities and in cows with milk fever (MF). The blood OPG and TRAP5b concentrations, as well as the ratio of OPG to TRAP5b (O/T), were measured beginning 3 weeks before (-21 d) and over a few days after calving in 49 Holstein Friesian cows at first (n = 8), second (n = 17), third (n = 12), and fourth or greater (n = 12) parities. The ratio of O/T at -21 d to O/T at calving (PreCOT) was also calculated. In the third and greater parities, seven cows developed MF (non-MF, n = 17). Regardless of the development of MF, the serum OPG started to decline in the last week of gestation only in the cows entering the second lactation, while the blood TRAP5b increased at calving in all cows. O/T decreased toward parturition only in multiparous cows. The decrease in O/T at caving was less pronounced in MF cows. PreCOT was negatively correlated with lactation number only in multiparous cows (n = 41, ρ = -0.50, P < .01). This study implied that OC differentiation toward calving was tapered in cows with advanced parities, and these indexes predict the risk of MF.

Adequate response of plasma 1,25-dihydroxyvitamin D to parturition in paretic (milk fever) dairy cows.

The concentration of 1,25-dihydroxyvitamin D was measured by means of a radioactive receptor assay in the plasma of cows during the period immediately prior to, during, and following parturition. Nonparetic cows showed initially a slight decrease in plasma 1,25-dihydroxyvitamin D which was followed by a significant increase during parturition and 2 days postpartum. The highest concentration achieved in the control or nonparetic cows was 100 picograms per milliliter. In the paretic animals the plasma 1,25-dihydroxyvitamin D concentration increased sharply during the day preceding calving and reached a maximum of 200 picograms per milliliter at parturition. This level was maintained during the ensuing 2.5 days. These results demonstrate that parturient paresis cannot be the result of insufficient synthesis or secretion of 1,25-dihydroxyvitamin D.

Evaluation of prepartum serum cholesterol and fatty acids concentrations as predictors of postpartum retention of the placenta in dairy cows.

OBJECTIVE: To identify serum biochemical and hematologic variables, as measured in the week before parturition, that predict postpartum retention of the placenta (RP) in dairy cows. DESIGN: Retrospective cohort study. ANIMALS: 1,038 cows in 20 commercial dairy herds. PROCEDURES: Serum concentrations of fatty acids (FAs), beta-hydroxybutyrate, cholesterol, glucose, urea, and calcium and blood leukocyte, neutrophil, lymphocyte, monocyte, and eosinophil counts were determined. These variables were evaluated for an association with development of RP by use of a multivariate logistic regression model. Parity, season of parturition, existence of twins or dystocia, body condition score, and vitamin E treatment were included in the model as covariates. RESULTS: High serum concentrations of cholesterol and FAs were associated with an increased odds of RP. There was a 5% relative increase in the odds of RP for each 0.1 mmol/L increase in cholesterol or FAs concentration in the week before parturition. Season of parturition and twinning were also identified as risk factors. CONCLUSIONS AND CLINICAL RELEVANCE: These associations indicated that prepartum energy metabolism contributes to the development of RP. Serum concentrations of cholesterol and FAs may be useful to identify cows with a metabolic abnormality or energy imbalance that might predispose them to RP and should be interpreted in conjunction with clinical risk factors such as twinning, dystocia, or parturient paresis.

Effect of intravenous calcium borogluconate and sodium phosphate in cows with parturient paresis.

Thirty cows with parturient paresis were divided into three groups of 10. All the cows were given 500 ml of a 40 per cent calcium borogluconate solution intravenously over a period of 10 minutes, and 20 were also given 500 ml of a 10 per cent solution of sodium phosphate intravenously; in 10 of the cows this solution was administered over a period of 10 minutes immediately after the calcium borogluconate solution, and in the other 10 cows 200 ml of the solution was administered rapidly and the remaining 300 ml was added to 10 litres of sodium chloride and glucose solution and infused slowly over six hours. There were no significant differences between the groups with respect to the outcome of the treatments; six or seven of the cows in each group stood within eight hours of the treatment. There were no significant differences between the changes in serum calcium concentrations among the groups. The mean concentrations of inorganic phosphorus in the groups given sodium phosphate were increased above the normal range initially, but after eight hours there were no significant differences between the groups in terms of the numbers of cows that were hypophosphataemic. There were no significant differences between the three groups with respect to changes after treatment in the serum concentrations of magnesium or parathyroid hormone.

[Efficacy of oral calcium and/or sodium phosphate in the prevention of parturient paresis in cows]. / Untersuchungen zur Prophylaxe der Gebärparese mit Kalzium und Natriumphosphat per os.

The goal of this study was to investigate the efficacy of calcium chloride, sodium phosphate or a combination of these two substances administered orally immediately postpartum for the prevention of parturient paresis in cows. Thirty-two cows that had had parturient paresis at the previous calving, and in which serum biochemistry had shown hypocalcaemia and hypophosphataemia, were used in the study. The cows were transferred to the Department of Farm Animals, University of Zurich, five days before their expected due dates. On a randomized trial, the cows were given calcium chloride, sodium phosphate, both substances or no treatment (controls) via a stomach tube immediately postpartum and 12 hours later. The cows were monitored for 96 hours during which time blood was collected on a regular basis for the determination of total calcium, ionized calcium, inorganic phosphorus and magnesium concentrations. Of the 32 cows treated, 19 (59%) had parturient paresis and 13 (41%) did not. The incidence of parturient paresis did not differ significantly among the groups although there was a tendency for a lower incidence in cows treated with both calcium chloride and sodium phosphate. The various treatments had no apparent effect on serum calcium concentration. The concentration of inorganic phosphorus increased significantly in cows treated with sodium phosphate compared with the controls. The results of this study showed that cows treated with both calcium chloride and sodium phosphate orally tended to have a lower incidence of parturient paresis. Further investigation into multiple administrations of oral calcium chloride and sodium phosphate, started before parturition, for the prevention of parturient paresis is required.

Influence of novel intravenous complex solution of Ca, Mg and phosphates on blood biochemical parameters of healthy and paretic cows.

The article describes the dynamics of changes in blood concentrations of the active substances present in the solution after its infusion to healthy cows in comparison to NaCI solution as well as the response of paretic cows to treatment with the new complex solution. Cows received a dose of 400 ml of A1 solution (containing 8.4 g of Ca2+) intravenously. In healthy cows the average calcium concentration in blood serum prior to the test was 2.52 +/- 0.08 mmol/l while 15 min. after the infusion the concentration rose to 3.10 +/- 0.08 mmol/l (p < 0.05) and magnesium concentration rose from 0.61 +/- 0.05 to 1.39 +/- 0.08 mmol/l (p < 0.05). This experiment showed that elevated concentration of non-organic phosphates persisted 1 hour after infusion (p < 0.05). In the second phase of efficacy evaluation of the novel preparation A1 on paretic cows the intravenous injection of 1 ml/kg of body weight of A1 solution increased calcium concentration up to almost normal level (p < 0.05). The level of magnesium in serum 1 h after injection was statistically significantly higher by 63% (p < 0.05) and reached the physiologically normal concentration. 1 h after the infusion of test solution the level of phosphate was higher by 13% (p > 0.05). The rise was statistically not significant. Even though A1 solution undoubtedly produced an increase in glucose concentration in the blood serum, due to wide dispersion of individual measurements and high standard deviation the increase (p > 0.05) in glucose concentration was found insignificant. Most of the treated paretic cows rose within 1-6 h after infusion of 400 ml of solution A1. No relapses were observed. A combination of different salts of calcium and magnesium, non-organic phosphates and glucose with analeptic substance mixed in one solution (A1 solution) administered at a dose of 1 ml/kg of body weight raises concentrations of essential macroelements in blood serum of cattle and promotes improvement of paretic cows condition.

Preventive effect of mildly altering dietary cation-anion difference on milk fever in dairy cows.

In the present study, we examined whether mildly altering dietary cation-anion difference (DCAD) contributes to the prevention of milk fever in dairy cows. Thirty multiparous cows and ten primiparous cows (heifer group) were used in this study and the multiparous cows were randomly divided into three groups of ten animals each (anion, non-anion and control groups). The cows in the anion group were given supplemental salts that slightly lowered DCAD. These salts consisted of 115 g of CaCO3, 42 g of CaHPO4, 65 g of MgSO4 x 7 H2O and 80 g of CaCl2 x 2 H2O as a daily dose for each cow, using a catheter from 21 days before the expected date of parturition until parturition. The cows in the non-anion group were given only the same Ca, Mg and ip supplement but no sulfate and chloride salts as that in the anion group. The cows in the control and heifer groups were not given any additional supplement. The incidence of hypocalcemia in the anion group decreased to approximately half of those in the non-anion and control groups, while the heifer group did not develop hypocalcemia at all. In addition, the number of days spent for the treatment of hypocalcemia and the number of drug bottles (calcium borogluconate solution) used for the treatment decreased to less than half in the anion group compared with those in the non-anion and control groups. At parturition, the serum Ca concentration in the control (6.2 +/- 1.9 mg/dl, mean +/- standard deviation) and non-anion groups (6.4 +/- 1.7 mg/dl) were significantly lower than that in the heifer group (8.3 +/- 0.4 mg/dl), and the level in the anion group was intermediate (7.3 +/- 1.3 mg/dl). The change in ionized Ca concentration was almost the same as that in serum Ca concentration, but only the concentration in the anion group tended to increase slightly from a week before parturition and was significantly higher than that in all other groups three days before parturition. Urinary pH in the anion group was maintained at a mildly acidic level (6.8-7.0) for the last two weeks before parturition, compared with those in the control (7.3-7.5) and non-anion groups (7.9-8.1), and similar to that in the heifer group (6.3-7.3). The urinary Ca excretion was the highest in the anion group among all groups during the prepartum period. There were no specific changes in the excretion of parathyroid hormone and 1,25-dihydroxyvitamin D in all groups of multiparous cows while the levels of these hormones remained low in the heifer group throughout the experimental period. The data in the present study indicates that the administration of anion salts that slightly lowered DCAD in the preparum period was effective for preventing milk fever in multiparous cows. Safe and mild metabolic acidosis induced by the anion salts could be evaluated by urinary pH (6.8-7.0), and might increase the responsiveness to Ca requirement at parturition through some complex mechanisms unrelated to the excretion of Ca-related hormones. In addition, it was clarified that primiparous cows have a high potential to respond to sudden Ca demand unrelated to hormone excretion, and their Ca metabolism was in some respects similar to that in multiparous cows fed anion salts. Therefore, manipulating mildly DCAD is expected to be an effective, safe and natural method for preventing milk fever in dairy cows.

Clinico-pathological findings in peripartum dairy cows fed anion salts lowering the dietary cation-anion difference: involvement of serum inorganic phosphorus, chloride and plasma estrogen concentrations in 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 clinico-pathological constituents in serum and urine, which might be related to milk fever, were analyzed using stored samples from the previous study to identify clinico-pathological parameters for easily evaluating the efficacy of lowering DCAD and to further investigate the mechanism by which lowering DCAD prevents milk fever. Among the parameters analyzed in the present study, inorganic phosphorus (iP) was involved in milk fever because the serum concentration and urinary excretion of iP were significantly higher in the group of primiparous cows (heifer group), which did not develop hypocalcemia, than those in other groups of multiparous cows. Serum chloride concentrations in the heifer group and the group of multiparous cows fed anion salts (anion group) tended to remain higher than those in other control groups of multiparous cows suggesting that serum chloride concentration may be utilized for evaluating the status of metabolic acidosis and the efficacy of lowerng DCAD in dairy cows fed anion salts. In addition, plasma estradiol-17beta concentration in the heifer group tended to be lower at parturition compared with that in other multiparous groups suggesting that estrogen known as a potent inhibitor of bone resorption may be involved in developing milk fever.

Milk fever and alert downer cows: does hypophosphatemia affect the treatment response?

The purpose of this prospective cohort study was to identify factors that place a dairy cow with uncomplicated milk fever (MF) at significant risk of becoming an alert downer cow (ADC) and to verify if these factors could be used to predict treatment outcome. Recumbent MF cows were examined before treatment and 52 were excluded due to complications. In all, histories and pretreatment serum samples were taken and the serum of 86 cows was analyzed for electrolyte levels (calcium, phosphorus, magnesium, and potassium). In total, 36 of the 86 samples were from ADCs and 50 from animals that responded to MF treatment (MFT). A binary-two-factor logistic model determined that a MF cow with a phosphorus pretreatment level of > or = 0.9 mmol/L was 12 times more likely not to become an ADC than one with a phosphorus level < 0.9 mmol/L (CI: 6.3,23.1). Also, a binary multivariable logistic regression analysis showed that a MF cow with a pretreatment calcium level > or = 1.7 mmol/L was 14 times more likely to become an ADC than one with a serum level < 1.7 mmol/L (CI: 2.0,98). Age and the other serum electrolytes were not statistically significant risk factors at the 0.05 level. The rigorous pretreatment examination and stringent adherence to protocol reduced ADC misclassification and fostered the strong association between single factor serum phosphorus levels and ADCs. By using a cutoff level of serum phosphorus at > or = 0.9 mmol/L, a practitioner could correctly predict that 95% of the MFs would not become ADCs and, therefore, this level would be a useful pretreatment predictor.

Effects of a sustained release formulation of 1,25-dihydroxyvitamin D3-glycosides for milk fever prevention on serum 1,25-dihydroxyvitamin D3, calcium and phosphorus in dairy cows.

Milk fever (MF) is a metabolic disease in dairy cows around parturition. The clinical lead sign is muscular paresis leading in severe cases to paralysis of the affected animal. Multiparturient animals of high performing dairy breeds are most likely to be affected and have a high probability of recurrence. An acute drop in blood calcium levels causes the disease when the demand for calcium at the onset of lactation exceeds the ability to replete blood calcium levels through mobilization from bone and intestinal uptake. With the understanding of the underlying mechanism, calcium supply management and vitamin D supplementation became prime candidates for MF prevention and therapy. Several strategies have been developed for MF prevention. Application of the active form of Vitamin D, 1,25(OH)2D3, was found to prevent MF effectively. In order to prevent a delayed hypocalcemia, which was occasionally seen after stopping the treatment with 1,25(OH)2D3, a new approach was chosen by applying Solanum glaucophyllum extract (SGE), which contains 1,25(OH)2D3-glycosides, as instant-release (irSGE) in combination with slow-release (srSGE) tablets. In a first study, non-lactating cows were treated with a single bolus of either synthetic 1,25(OH)2D3, irSGE, or srSGE and the results were compared to a control group without treatment. Blood serum levels of 1,25(OH)2D3 (1,25D), calcium (Ca), phosphate (P) and magnesium (Mg) were followed for 11days and the area under the curve (AUC) was calculated. Calcium and phosphate excretion in urine were determined during 15days. While serum concentration of 1,25(OH)2D3 was back to pre-treatment level in the irSGE, srSGE and 1,25(OH)2D3 treated group within 3days, calcium and phosphate levels remained elevated for up to 9days. AUC of serum 1,25(OH)2D3 was 2.89 (1,25D), 3.13 (irSGE) and 4.21 (srSGE) times higher than control. Serum calcium levels were 1.07* (for 1.25D); 1.08* (for irSGE) and 1.12* (for srSGE) times higher than control. Serum phosphate levels were 1.20* (for 1,25D); 1.30* (for irSGE) and 1.41* (for srSGE) times higher than control, with * p<0.05. In a second field study calving cows treated with one bolus containing ir- and sr- tablets of SGE were compared to an untreated control group and to a group treated with 4 boli of commercial calcium salts. As a result, calcium serum levels increased (+19% compared to baseline) around calving after treatment with the single bolus of SGE. The single bolus of SGE lead also to an increase of serum phosphate (+31% compared to baseline). These calcium and phosphate increases were statistically significant (p<0.001) 0-24h after calving compared to the control group and to the group treated with calcium salts. The sample size of the study was too small to draw a conclusion on the effect on MF prevention. In conclusion, application of a single bolus of a SGE extract lead to an increase of serum calcium and phosphate for up to 9days and may thus have the potential to prevent a hypocalcemia and -phosphatemia, an important cause for clinical milk fever.

Parturition and hypocalcemia blunts calcium signals in immune cells of dairy cattle.

The stress of parturition in the dairy cow is associated with increased susceptibility to infectious disease. During the periparturient period the demands for calcium are increased; these increased demands for calcium can result in subclinical or clinical hypocalcemia. Periparturient cows also experience significant immune suppression. Because intracellular calcium signaling is a key early feature in immune cell activation, we have hypothesized that the increased demand for calcium in periparturient cows may adversely affect intracellular calcium stores of immune cells. This reduction in intracellular calcium stores in immune cells could blunt intracellular calcium release following an activating stimulus, contributing to the immune suppression seen in these animals. To test this hypothesis, peripheral mononuclear cells were obtained from 27 multiparous dairy cows spanning a period of 2 wk before and 2 wk after parturition. Following activation of these cells by anti-CD3 antibodies plus secondary antibodies, intracellular calcium release from intracellular stores was measured. The intracellular calcium released in response to the activation signal declined as calcium demand for lactation became more intense and recovered as plasma calcium normalized. Intracellular calcium stores in peripheral mononuclear cells, estimated by pretreating cells with pervanadate and ionomycin, significantly decreased at parturition and returned to normal levels as the cows' blood calcium returned to normal levels. Hypocalcemia, which is common in periparturient dairy cows, is associated with decreased intracellular calcium stores in peripheral mononuclear cells. Our data suggest that this is the cause of a blunted intracellular calcium release response to an immune cell activation signal. It is concluded that intracellular Ca stores decrease in peripheral blood mononuclear cells (PBMC) before parturition and development of hypocalcemia. This suggests that systemic calcium stress precedes measurable hypocalcemia, particularly in cows that will develop milk fever. Therefore, PBMC intracellular Ca stores are a more sensitive measure of calcium stresses in transition cow. This decrease in PBMC intracellular Ca stores before parturition and the development of hypocalcemia contributes to periparturient immune suppression.

Relationship between blood calcium level and ST peak interval of electrocardiographic variables in peripartum Holstein cows.

The association between blood calcium levels and electrocardiographic variables was compared in 137 normal parturient and 36 peripartum recumbent Holstein cows to determine whether hypocalcemia in peripartum dairy cows can be rapidly diagnosed using electrocardiograph. Inverse of STc (ST peak interval/SS interval(0.5)) and blood ionized calcium or serum calcium concentrations were strongly correlated, and both correlation coefficients were 0.81 (P<0.001). The 95% prediction interval indicated that cows with STc >0.385 ± 0.001 sec are very likely to be hypocalcemic (blood ionized or serum calcium concentrations of <0.9 mmol/l or <7.5 mg/dl, respectively). These findings indicate that hypocalcemia in parturient cows can be non-invasively estimated using the STc.

Correlation of different biochemical parameters in blood sera of healthy and sick cows.

Metabolic diseases of cows represent the leading internal pathology in Lithuania in terms of incidence and economic impact. This paper summarizes the mineral metabolic state of milk cows, and details the influence of feeding on serum levels of calcium, nonorganic phosphorus, magnesium, potassium, sodium, glucose, total protein, and urea quantity, and correlationd among them, in healthy dry or post-calving cows as well as in cows with osteomalacia and milk fever after calving. There was less pronounced hypocalcaemia and there were only minor changes in phosphorus, magnesium, potassium and sodium in the serum of healthy dry and post-calving cows that had silage and mineral-vitamin supplements, compared to cows that did not have supplements and silage. There was a fall in calcium and phosphorus (to 1.89 +/- 0.12 mmol/L and 0.71 +/- 0.06 mmol/L, respectively) in the blood of cows sick with milk fever after calving, while levels of magnesium and potassium were increased. The correlation between calcium and phosphorus was r = 0.6993, p < 0.001 in the serum of sick cows. There was a fall in calcium, phosphorus and magnesium (to 1.86 +/- 0.46 mmol/L, 0.75 +/- 0.37 mmol/L, and 0.60 +/- 0.19 mmol/L, respectively) and an increase in sodium level (to 158.90 +/- 19.30 mmol/L) in the blood of cows with osteomalacia in comparison with healthy cows.