The influence of short-term anion salt exposure on urine pH and on resistance to experimentally induced hypocalcaemia in cows

The objective of this experiment was to induce acidification by anion salt supplementation for 2 days or 10 days and to study the prophylactic effects of such supplementation in preventing hypocalcaemia in cows. It was further attempted to monitor the extent to which any effect on the calcium-regulating mechanisms would persist following a 10-day period of acidification with anion salts. Study animals were three untreated control cows and three cows supplemented with ammonium chloride and ammonium sulphate in their ration for 2 days or 10 days through the rumen cannula. The basic ration of hay was dominated by Urochloa spp. The pH of the urine of the control cows was around 8.00 throughout the experiment and was considered normal. Anion-supplemented cows produced urine with a daily mean pH between 5.5 and 7.0, possibly due to anion salt exposure. The ability to withstand hypocalcaemic challenges was tested by a standardized intravenous infusion of disodium ethylenediaminetetraacetate (Na2EDTA). The calcium regaining time (CRT), expressed as time spent to reach 1.00 mmol/l of ionized calcium during recovery from the EDTA-induced hypocalcaemia, was used to compare cow responses. In the control cows the unexpectedly short CRT, especially during the weekly EDTA tests, could be a result of the repeated induced episodes of hypocalcaemia caused by the EDTA infusions. The improved CRT in the anion-supplemented cows may thus be interpreted as the combined effect of the repeated hypocalcaemic episodes due to EDTA infusions and probably the effect of anion-induced metabolic acidosis on endocrine-regulated calcium homeostatic mechanisms. The effect of anion salt exposure for 10 days on the improvement of calcium-regulating mechanisms was not clear due to the unexpected improvement of CRT that was exhibited by the untreated control cows as well. An on-farm trial of the effect of a 2-day or 10-day anion exposure of dry cows on calcium-regulating mechanisms is suggested.     

Serum calcium response following oral zinc oxide administrations in dairy cows

Six non-pregnant cows were allocated into 3 groups. Group 1 comprised a pair of lactating cows, whereas groups 2 and 3 each comprised a pair of non-lactating cows. The cows in groups 1 and 2 were dosed intraruminally by stomach tube with zinc oxide at 120 mg Zn per kg of bodyweight at weekly intervals for a period of 33 days. Each cow received a total of 4 doses of zinc oxide. Group 3 served as non-treated control group. Blood samples were collected from all 6 cows daily. Serum was analysed for concentration of calcium. Within 12-24 h of each zinc oxide administration the serum calcium of the lactating cows dropped dramatically indicating the existence of an antagonistic effect between Zn and Ca. The first Zn induced hypocalcaemic episode in the lactating cows was followed by a rise in serum calcium to a level above the pre-dosing level and above the mean value of the control group. The depth of the hypocalcaemic response decreased with the number of zinc oxide dosings. This effect was explained as a response from the stimulation of the calcium homeostatic mechanisms. In the Zn dosed non-lactating cows responses were similar but less clear. The perspective of these findings is discussed in relation to resistance towards parturient hypocalcaemia.     

Lipid mobilization in parturient cows.

The blood levels of calcium, inorganic phosphorus, magnesium, glucose and NEFA were studied in cows at the time around partus. Eight of 16 cows developed hypocalcaemic paresis. Besides hypocalcaemia the paretic cows showed lower levels of inorganic phosphorus and higher levels of glucose and NEFA than non-paretic cows 24 hrs. post partum. It is known that lipolysis is associated with uptake of calcium in adipose tissue. The calcium content in perirenal adipose tissue was however lower in paretic cows than in non-paretic parturient cows and lactating cows slaughtered 3–5 months after calving. The calcium content in omental adipose tissue was about the same in all 3 groups. Despite increased lipolysis the calcium content in adipose tissue is thus not increased in cows suffering from parturient paresis.     

Protein expression in dairy cows with and without subclinical hypocalcaemia

AIM: To determine differences in plasma proteomic profiles between healthy cows and those with subclinical hypocalcaemia within 12 hours after calving, and thereby explore the underlying biological mechanism of subclinical hypocalcaemia in dairy cows.

METHODS: Plasma samples were collected within 6 hours of calving from Holstein cows on a farm in Heilongjiang, China; 32 with subclinical hypocalcaemia (plasma calcium concentration 1.38–2.00?mmol/L and no clinical signs) and 59 control cows (plasma calcium concentration 2.10–2.8?mmol/L). Plasma samples were applied to weak cationic exchange protein chips for protein profiling by surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS), and the data were analysed using the PBS-IIC system. The amplitude of peaks for the two groups were compared using the Wilcoxon sum-rank test, and the mass-to-charge ratio of the peaks that differed was used to identify peptide fragments using the Swiss-Prot protein database.

RESULTS: Seven peaks were identified in the subclinical hypocalcaemia group that differed from those of the control group (p<0.001), that represented six unique proteins. Expression of serum albumin, fibrinogen alpha chain, amyloid beta A4 proteins and neurosecretory protein VGF were increased, and expression of apolipoprotein A-II and serum amyloid A proteins were decreased in the subclinical hypocalcaemic cows compared with control cows.

CONCLUSION: Use of SELDI-TOF-MS technology can effectively identify differences in plasma protein expression patterns in cows with subclinical hypocalcaemia. Neurosecretory protein VGF and amyloid beta A4 protein might represent useful biomarkers for diagnosis of subclinical hypocalcaemia.     

Effect of 1,25 dihydroxyvitamin D3 on the calcium and magnesium metabolism of lactating cows

Administration of 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) to lactating dairy cows resulted in increased dietary calcium absorption and elevated concentrations of plasma calcium. Dietary magnesium absorption was unaffected by 1,25(OH)2D3 however, plasma magnesium concentration was depressed. Injections of 1,25(OH)2D3 were effective in elevating plasma calcium concentrations in both normal and hypomagnesaemic cows. This indicates a potential use for 1,25(OH)2D3 to prevent and treat hypocalcaemic cows with or without concurrent hypomagnesaemia.     

Calcium effects on renal conservation of magnesium in cows

Urinary excretion rates for Ca, Mg and P were compared to simultaneous measurements of the same elements in blood plasma of cows recovering from moderate hypocalcaemia post partum and in 1 animal during recovery from hypocalcaemia induced by fasting.The renal conservation of Mg was found to be increased during the hypocalcaemia, theoretical tubular reabsorption rates being correlated to plasma Ca with r = –0.7, P < 0.001. The relationship was observed in the plasma Ca range from about 2.0 to 2.6 mmol/1.The findings may explain the occurrence of hypermagnesaemia in hypocalcaemic cows and the delay in the development of hypo-magnesaemia seen in cows with fasting-induced hypocalcaemia.     

Effect of pre-calving zeolite, magnesium and phosphorus supplementation on periparturient serum mineral concentrations

The objective of this study was to test whether supplementing dry cow rations with phosphorus (P) and magnesium (Mg) would interfere with the beneficial effect of zeolite supplementation on the periparturient blood calcium (Ca) concentration in dairy cattle. Three groups (A-C) of 10 Danish Jersey cows were each given the following daily supplements from 2 weeks before the expected date of calving until actual calving: group A: zeolite, monoammonium phosphate, standard dry cow mineral and vitamin mix, containing 61g magnesium phosphate; group B: zeolite, standard mineral and vitamin mix without the magnesium phosphate and group C: standard mineral and vitamin mix, monoammonium phosphate. All cows in group B had an apparently less variable serum calcium concentration around calving with no cases of milk fever and no subclinical hypocalcaemia or hypomagnesaemia recorded. In contrast, a parturient drop in blood Ca was seen in group A as well as group C. In group A, one cow was hypocalcaemic at calving, and developed milk fever. In group C, 12 blood samples, representing six cows, were hypocalcaemic, and three of these cows were treated for milk fever. All groups remained normomagnesaemic and there were no significant differences in blood Mg across groups. In conclusion, the combined P and Mg supplementation in addition to zeolite supplementation did not increase the serum Mg level (forage Mg 16.9g/day; 0.21% of DM). Combined P and Mg supplementation reduced the zeolite-induced hypophosphataemia but also reduced the stabilising effect of zeolite on parturient serum Ca.     

Immunohistochemistry of ultimobranchial thyroid carcinomas in seven slaughtered cows and one bull.

Eight thyroid gland epithelial tumors were found in 7 cows and 1 bull in a retrospective study of thyroid gland lesions in slaughtered cattle. All tumors were classified as ultimobranchial thyroid carcinomas based on morphology and immunohistochemistry. All tumors consisted of solid sheets and nests of polygonal to oval epithelial cells, with more sparsely dispersed colloid-filled follicles. Connective tissue separating nests of epithelial cells varied from delicate fibrovascular stroma to dense collagenous stroma. Fusiform epithelial cells with rare neural fibers and ganglion cells were present in 1 tumor. Cells within solid areas of these tumors were immunoreactive for calcitonin, calcitonin gene-related peptide, neuron-specific enolase, and synaptophysin. Colloid and follicle cells were immunoreactive for thyroglobulin. Few follicle cells also were reactive for calcitonin gene-related peptide. Neoplastic cells invaded the fibrous capsules in all 8 cattle. These tumors represented proliferation of a mixed population of undifferentiated cells, C cells, and thyroid follicular epithelial cells, presumably derived from the thyroid ultimobranchial bodies. These ultimobranchial carcinomas in slaughtered cattle are comparable to ultimobranchial tumors described in dairy bulls and the intermediate type of thyroid gland carcinomas (mixed thyroid medullary carcinomas) described in human beings.     

Further studies on theophylline-induced hypocalcaemia in sheep.

The effects of various doses of theophylline on the plasma levels of calcium, inorganic phosphorus and magnesium were studied in sheep. The dose of 0.25 mg/kg/min. during 40 min. produced a hypocalcaemia, which was abolished by thyroidectomy. At a higher dose, 1.0 mg/kg/min., thyroidectomy reduced, but did not block, the hypocalcaemic effect of theophylline. Young sheep showed a greater calcium response than did adult ewes. The decrease in plasma calcium was in no case accompanied by hypophosphataemia. Plasma magnesium decreased in all groups after 1.0 mg/kg/min. and was not influenced by thyroidectomy. Theophylline increased the urinary excretion of calcium and magnesium. It is concluded that theophylline-induced hypocalcaemia in sheep is in part a result of increased calcitonin secretion but that also other mechanisms must be involved, e. g. a direct effect of theophylline on bone.     

Interaction between the levels of hormones and minerals in sera of healthy and sick cows

An interaction between blood levels of parathyroid hormone, calcitonin, 1.25-dihydroxycholecalciferol and levels of calcium, phosphorus and magnesium was examined in 85 cows, which included healthy cows and cows with ostemalacia, mastitis and paresis. Levels of parathyroid hormone (PTH) and calcitonin were determined in vitro using IMMULITE analyser (Diagnostic Products Corporation, USA), by means of immunometric assay. Levels of vitamin D were measured using the enzyme linked immunosorbent assay (ELISA). Levels of calcium, phosphorus and magnesium were determined using the automated Eos-Bravo analyser (Hospitex Diagnostics, Italy) with HOSPITEX reagents. The lowest blood levels of calcium (1.38 +/- 0.18 mmol/L) and phosphorus (0.65 +/- 0.12 mmol/L) were found in cows with parturient paresis. Decreased blood levels of phosphorus and magnesium were also determined in cows with osteomalacia. For cows with parturient paresis, which received a mineral supplement, the average serum level of calcium was by 20.7% higher than the level found in those which did not receive a supplement, and the level of phosphorus was by 23.6% higher, however, these levels remained low. The blood level of parathyroid hormone ranged from 3.47 to 5.20 pmol/L in healthy cows and from 3.95 to 15.21 pmol/L in sick cows. The highest and statistically significant increase in blood PTH level (up to 18.31 +/- 1.88 pmol/L) was found in cows with parturient paresis. The blood level of PTH correlated inversely with the level of calcium in cows with osteomaliacia (r = -0.89) and in cows with parturient paresis (r = -0.49 and r = -0.61, respectively). The serum level of calcitonin ranged from 1.46 pmol/L to 2.40 pmol/L in healthy and sick cows and the difference was not statistically significant. Lower serum levels of vitamin D were found in heifers-in-calf and in cows with mastitis. A clear correlation between levels of calcitonin, vitamin D and macronutrients was not found.     

Factors affecting the success rate of treatment of recumbent dairy cows suffering from hypocalcaemia

We aimed to investigate the ratio of accompanying diseases in cows suffering from clinical hypocalcaemia and their influence on cure rate. In five veterinary practices in different regions of Germany, all recumbent cows around parturition were included in the study for a period of 1 year. After recording the case history a clinical examination was done and a serum sample was taken to measure the concentrations of calcium and phosphorus magnesium, beta-hydroxybutyrate, total bilirubin, cholesterol, urea, and the activities of ASAT, CK, and GLDH. Only cows with hypocalcaemia entered the statistical analysis. Hypocalcaemia was the major cause of recumbency in cows of the second lactation or elder. Muscle damage was the second frequent diagnose in recumbent cows and the major concomitant disease in hypocalcaemic cows. The overall cure rate was between 89.4% and 94.8%. Calcium and phosphorus concentrations did not have an influence on cure rate. Non-cured cows had higher serum activities of CK (p<0.043) and ASAT (p<0.006). Nevertheless, the activities of CK and ASAT were no good predictors of treatment failure because of their low specificity and the high cure rate of the cows in the five practices.     

Transient hypoparathyroidism following successful treatment of hypercalcaemia of malignancy in a dog

A 4-year-old, entire female, English Cocker Spaniel was presented for treatment of lymphosarcoma and secondary hypercalcaemia. After induction chemotherapy the dog became severely hypocalcaemic and showed signs of weakness, muscle fasciculation and facial pruritus. Hormone analysis confirmed inadequate production of parathyroid hormone. Although hypocalcaemia has been previously reported as a component of tumour lysis syndrome, it has not been associated with transient parathyroid hormone deficiency.     

Localization and expression of gastrin-releasing peptide (GRP) in the bovine cervix

Gastrin-releasing peptide (GRP) has been suggested as a novel regulatory peptide in the female reproductive tract but the presence of GRP and GRP mRNA in the non-neurogenic tissue of the cervix has not yet been clarified. In the present study, immunohistochemistry and in situ hybridization were used to reveal the distribution of GRP immunoreactivity and expression of GRP mRNA in the bovine cervix. The cervixes from 21 non-pregnant and 20 pregnant cows, and 6 fetuses were used in the study. In the fetus, adult non-pregnant and pregnant specimens, GRP and GRP mRNA were predominantly detected in the luminal epithelial cells of basal areas of peripheral regions of the cervix. Positive staining of GRP in the epithelial cells of the cervix was first detected in the CRL 37 cm of the fetus. During the estrous cycles, the staining intensity of GRP in the epithelial cells was stronger in the follicular phase than in the luteal phase. During the early gestational period, GRP immunoreactivity was detected at relatively similar intensity to the follicular phase. In situ hybridization results ascertained the expression of GRP mRNA in the superficial epithelial cells of the cervix of non-pregnant and pregnant cows. The results suggest that GRP may be important both in the development of the fetal cervix and secretory activity of the epithelial cells of the cervix.     

Pathological changes in the myocardium of hypocalcaemic parturient cows

Three parturient cows in lateral recumbency which were moaning and had tachycardia, arrhythmia and dyspnoea were thoroughly examined. They were hypocalcaemic (0.70-1.27 mmol/litre) and were euthanized within four days, because they failed to respond to calcium treatment. By light microscopy the most characteristic pathological changes in the heart were necrotic changes scattered throughout the myocardium. Electron microscopy revealed abnormalities in the myocytes which were characterised by focal myofibrillar lysis, irregular splitting, streaming and spreading of the Z band and myofibrillar disarray.     

Decreases in serum apolipoprotein B-100 and A-I concentrations in cows with milk fever and downer cows

Milk fever occurring during the peripartum period has been suggested to be caused by fatty liver developed during the nonlactating stage because diseased cows have increased serum concentrations of non-esterified fatty acids (NEFA) and show hepatic lipidosis. In cows with fatty liver and related diseases such as ketosis, serum concentrations of apolipoprotein (apo) B-100 and apoA-I are decreased. The purpose of the present study was to examine whether apoB-100 and apoA-I concentrations are similarly decreased in cows with milk fever. Apolipoprotein concentrations were also measured in cows with downer syndrome, which has been suggested to be related, at least in part, to milk fever. Compared with healthy cows during early lactation, apoB-100 and apoA-I concentrations were decreased in cows with milk fever and also in downer cows. In cows with milk fever, the decreases in apoB-100 and apoA-I concentrations were associated with increased NEFA and decreased cholesterol and phospholipid concentrations. However, in downer cows, serum lipid concentration changes were not as distinct as in cows with milk fever. These results, coupled with previous findings on the decreases in apoB-100 and apoA-I concentrations of cows with fatty liver-related diseases, suggest that fatty liver is involved in the development of milk fever and partly in that of downer cow syndrome.     

饲粮阴阳离子差对围产期奶牛血清生理生化指标及犊牛血清抗氧化指标的影响

本试验旨在研究围产前期饲粮阴阳离子差(DCAD)对产后奶牛血清生理生化指标及犊牛血清抗氧化指标的影响。试验采取单因子随机区组试验设计,选取2~4胎次、体重相近、预产期相近的围产前期(产前28天)中国荷斯坦奶牛20头,分为4组,每组5头。4组分别在围产前期饲喂DCAD为+262.31、+130.26、+78.51、+6.67 mmol/kg(干物质基础)的饲粮,阴离子盐的添加量分别为0、15.0、21.9、29.1 g/kg。试验期49 d,其中预试期7 d,正试期42 d。结果表明:降低奶牛产前饲粮DCAD能显著提高其血清钙含量(P0.05);显著降低尿液p H(P0.05);显著提高产后血清维生素D含量(P0.05);显著提高犊牛血清抗氧化能力(P0.05);但对奶牛血清肿瘤坏死因子、甲状旁腺素、降钙素和β-羟丁酸含量没有显著影响(P0.05)。因此,添加阴离子盐降低产前奶牛饲粮的DCAD可以促进其血清钙稳态,诱发机体轻度代谢性碱中毒从而减少产褥热的发病率,并提高犊牛的抗氧化能力。本试验条件下,15.0 g/kg(干物质基础)为阴离子盐最佳添加量。     

Factors affecting the hypocalcaemic response to protamine.

The hypocalcaemic response to protamine, as a measure of bone resorption rate, was studied in cattle and sheep. Three groups of calves were studied (1–3 days, 2–3 weeks and 4 months old). The oldest calves showed the greatest response, indicating a more rapid skeletal turnover in these animals, probably related to a higher growing rate. A group of cows was tested at 2 occasions, near parturition and 2–3 weeks after parturition. The decrease in plasma calcium was small near parturition, while a profound drop was obtained 2–3 weeks post partum, which supports earlier findings that bone resorption is more or less blocked in parturient cows. Young pregnant ewes showed the same degree of hypocalcaemia as adult pregnant ewes but the recovery was more rapid in the young animals. Young rams were given protamine before and after diethylstilboestrol treatment. No differences in calcium response were obtained. Varying plasma phosphate responses were obtained. A pronounced hyperphosphataemia was found in the adult pregnant ewes. Diethylstilboestrol caused hypermagnesaemia in the rams.     

Blood pathophysiological changes in sheep following a prolonged (18-hour) period of hypocalcaemia induced by Na2EDTA solution.

Six aged Merino ewes were used in an experiment in which five were infused with 4.7% Na2EDTA solution intravenously for 18 h at a rate designed to produce hypocalcaemia and maintain recumbency, and five with 0.9% sodium chloride solution at the same rate for the same period (four were infused at different times with both solutions). Blood samples were collected every 3 h and determinations made of plasma Ca, Na, K, Mg, and inorganic P (PiP), erythrocyte Na, K and Mg, and PCV. Three of the hypocalcaemic sheep took 36-64 h to regain their feet. Plasma Ca and K, and erythrocyte Na showed significant (all P less than 0.01) decreases in the group infused with Na2EDTA compared with the group infused with saline while PCVs were significantly (P less than 0.01) greater in the former group. The sheep model used could be suitable for the study of the effects of prolonged hypocalcaemia and recumbency in 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.     

Avian tibial dyschondroplasia: a comparison of the incidence and severity as assessed by gross examination and histopathology.

The assessment of avian tibial dyschondroplasia (TD) in many recent studies is based on examination of slices of the proximal tibiotarsus with the naked eye. This study examines the incidence and severity of TD in broilers under four different dietary regimes and compares the efficacy of naked eye assessment with histopathological examination. The diets contained reduced levels of calcium relative to phosphorus with adequate (diet 1) and high (diet 2) levels of vitamin D3 supplementation; a very low calcium diet (diet 3) and a standard diet (diet 4) were also included. Gross examination suggested that TD was present in 80 per cent, 79 per cent and 27 per cent of tibiotarsi from birds on diets 1, 2 and 4, respectively. However, histological examination indicated TD, correspondingly, to be present in 18 per cent, 39 per cent and 6 per cent of tibiotarsi. Some birds on diet 1 exhibited physeal changes consistent with mild hypocalcaemic rickets. Many of the bone extremities which, grossly, were considered dyschondroplastic (diets 1, 2 and 4), histologically were shown to have an extensive hypertrophied zone with poor metaphyseal vessel penetration, but no cellular or matrix changes consistent with TD. This cast doubt on the validity of earlier studies of TD which relied solely on naked eye examination. The birds on diet 3 all demonstrated histopathology considered consistent with severe hypocalcaemic rickets and the present study suggests that in three-week-old broilers a moderately hypocalcaemic diet with a high level of vitamin D3 will not cause rickets but will increase the incidence and severity of TD.(ABSTRACT TRUNCATED AT 250 WORDS)