Ceruloplasmin as an indicator of copper status in cattle and sheep.

The relationship between ceruloplasmin, a metalloenzyme with oxidase activity, and copper was investigated in cattle and sheep. The oxidase activity of ceruloplasmin correlated closely with the serum or plasma copper concentrations in cattle. The respective correlation coefficients were 0.83 and 0.60. In sheep serum, a correlation coefficient of 0.92 was obtained. In each instance, the relationship remained linear from the deficient to the high normal ranges of copper. Comparison of the linear regression relationships indicated the ceruloplasmin activity in bovine serum was statistically lower than the activity in bovine plasma (P less than 0.0001), through the intercepts from the regression lines of the two relationships were similar (P = 0.412). Comparisons of ovine and bovine serum-ceruloplasmin relationships indicated that a significant species difference was present. Ovine ceruloplasmin activity increased more rapidly as compared to the corresponding bovine activity over the range of copper concentrations investigated (P less than 0.0001). The intercept from the ovine regression relationship was also lower (P less than 0.0001). A correlation coefficient of 0.35 was observed between the serum ceruloplasmin activity and hepatic copper concentrations in cattle indicating that the mathematical relationship was not as well defined. Ceruloplasmin activity appears to correlate more closely with serum or plasma copper concentrations as compared to corresponding liver copper concentrations.     

Acute phase response in horses: changes in plasma cation concentrations after localised tissue injury

An acute phase reaction was elicited in four horses to which Freund's adjuvant was administered intramuscularly. The localised inflammation was accompanied by changes in the plasma concentrations of copper, iron and zinc. The plasma copper concentration, the plasma ceruloplasmin copper concentration and the ceruloplasmin oxidase activity in the plasma steadily increased to a maximum 24 days after the administration of the adjuvant. At this time, the plasma copper concentration was 2.2 micrograms/ml, a 90 per cent increase over the baseline concentration. The ratio of the concentration of plasma ceruloplasmin copper to plasma copper remained constant, indicating that the non-ceruloplasmin bound copper component of the plasma is also an acute phase reactant in the horse. The plasma zinc and iron concentrations decreased to 59 per cent and 30 per cent of their respective baseline concentrations and the severity of the inflammation appeared to influence the plasma concentrations of each metal. Weak correlations between the plasma fibrinogen concentration and the plasma copper and zinc concentrations of 25 horses with plasma fibrinogen concentrations of 5 g/litre or greater indicated that a single measurement of plasma copper concentration is not useful in the diagnosis of non-specific inflammatory disorders of the horse. However, the results suggest that the plasma copper concentrations in serial samples may be used to monitor the resolution of inflammatory disorders in the horse.     

Immunohistochemical demonstration of cytochrome oxidase in different parts of the central nervous system: a comparative experimental study

Cytochrome oxidase, the terminal enzyme of the electron transport chain, is a marker of the functional activity of the cell. In this study; localization of cytochrome oxidase in cerebrum, cerebellum, hippocampus, substantia nigra and choroid plexus of adult rats was investigated using immunohistochemical methods. Neural bodies were immunoreactive while neuroglial cells and axonal areas did not show significant immunostaining. The cerebral cortical substantia grisea region was stained almost homogeneously with cytochrome oxidase. In the cerebellar cortex, immunolabelling was more intense in the granular layer than the molecular layer. There was significant immunostaining in Purkinje cells. White matter, both in cerebrum and cerebellum, did not show immunoreactivity for cytochrome oxidase. Neurones in the hippocampus showed variable immunostaining; some of them were negative while others revealed high immunoreactivity. The neurones in substantia nigra were heavily labelled. Immunostaining for cytochrome oxidase in plexus choroideus epithelial cells was also remarkable. The morphological findings demonstrate the regions which most require and produce energy and reflect the differences in cellular activity in these parts of the central nervous system.     

Changes in enterocyte mitochondria associated with deficiency of copper in cattle.

Comparisons were made between specimens of intestinal mucosa from three young Friesian steers, killed when showing clinical signs associated with copper deficiency, and three controls that had received supplementary copper. Copper deficiency was associated with marked depletion of cytochrome oxidase in the epithelium of the duodenum, jejunum and ileum, and with partial villus atrophy in the duodenum and jejunum. Enterocytes from the duodenum, jejunum and ileum showed mitochondrial abnormalities ranging from slight swelling to marked localised dilation. Many of the mitochondria not affected by swelling had a distinctly condensed appearance. These changes are discussed in relation to the copper-responsive diarrhoea that affects a proportion of cattle suffering from copper deficiency.     

Copper status and enzyme, hormone, vitamin and immune function in heifers

A survey was conducted in 10 districts of Northern India to record the copper deficiency in soil, fodder and serum samples. Significant deficiency of copper was observed in soil, fodder and serum samples of heifers. The copper deficient animals were listless, showed depigmentation of skin, stiff gait and were anaemic and diahorric. The purpose of the study was to evaluate the magnitude of copper deficiency in Northern India and to examine the various haematobiochemicals, enzymes, vitamin and immune function which are affected by the deficiency of copper, so as to identify the parameters which can be of diagnostic importance in copper deficiency. Forty hypocuperemic heifers were selected from these areas and were randomly divided into two groups A and B. The heifers in group A were provided with mineral mixture containing copper sulphate and in group B without copper sulphate. Significant improvement (P<0.01) was observed in the haemoglobin (Hb), total leukocyte count (TLC) and total erythrocyte count (TEC) level at the 30th day of treatment in the animals of group A. Significant (P<0.01) improvement in serum. Ceruloplasmin (Cp) level was observed within 30 days of treatment, while significant (P<0.01) improvement in monoamine oxidase and liver cytochrome oxidase was observed at the 60th day of treatment in group A animals. Regarding hormones significant improvement was observed in T(3) and T(4), in the animals of group A within 60 days of treatment. The values of vitamin A and E showed significant (P<0.01) improvement within 30th days of treatment. The phagocytic activity of neutrophils against Candida albicans significantly (P<0.01) improved in group A within 60 days of treatment. Similarly significant improvement in superoxide dismutase activity in RBC was observed at the 30th day, and WBC and whole blood at 60th day in group A animals. Significant improvement in liver Cu level was observed at the 30th day of treatment, while in group B the liver Cu was significantly (P<0.01) depleted at the 60th day of experimentation. Additional Cu supplementation improved growth performance significantly in group A.     

Therapeutic management of copper deficiency in buffalo heifers: Impact on immune function

To evaluate the magnitude of copper deficiency in Northern India and to examine the various haematobiochemicals, enzymes, vitamins and immune functions affected by copper deficiency, and to identify the parameters which can be of diagnostic importance in copper deficiency, a survey was conducted in 12 districts of Northern India. Significant deficiency of copper was observed in soil, fodder and serum samples of buffalo heifers. Fifty hypocuperaemic buffalo heifers were selected from these areas and were randomly divided into two groups, A and B. The heifers in group A were provided with mineral mixture containing copper sulphate and in group B without copper sulphate. Significant (p < 0.01) improvement in serum ceruloplasmin level was observed within 30 days of treatment, while significant (p < 0.01) improvement in monoamine oxidase and liver cytochrome oxidase was observed at the 60th day of treatment in group A animals. Significant improvement was observed in T₃ and T₄, in the animals of group A within 60 days of treatment. The values of vitamin A and E showed significant (p < 0.01) improvement within 30 days of treatment. The phagocytic activity of neutrophils against Candida albicans significantly (p < 0.01) improved in group A within 60 days of treatment. Similarly, significant improvement in superoxide dismutase activity in red blood cells was observed at the 30th day, and in total leukocytes and whole blood at the 60th day in group A animals. Significant improvement in liver copper level was observed at the 30th day of treatment, while in group B the liver copper was significantly (p < 0.01) depleted at the 60th day of experimentation. Additional copper supplementation improved growth performance significantly in group A.     

日粮含铜量对生长猪血浆的几种微量元素浓度及酶活性影响的研究

<正> 前言 铜(Cu)是家畜不可缺少的一种微量元素。Mchargue(1926)最早证明了Cu对植物和动物的重要生物学作用。Brande(1967),Gipp(1973)及Wallace(1960)等人相继报导了在日粮中添加Cu对提高生     

Serum biochemical changes in calves with Johne's disease

SUMMARY Calves clinically affected with experimentally induced Johne's disease exhibited elevation of caeruloplasmin oxidase activity, and marked depression of α-mannosidase activity during the period when clinical signs of the disease were most prominent. Changes in serum copper levels and alkaline phosphatase activity were closely correlated with the elevation of caeruloplasmin oxidase activity, and depression of α-mannosidase activity. The pattern of these changes was similar to nutritional and metabolic changes described previously in acute infectious conditions in man and animals.     

The role of sulphide and sulphide oxidation in the copper molybdenum antagonism in rats and guinea pigs

The sulphide metabolism of rats fed molybdate up to levels of 1000 ppm molybdenum was examined and large decreases in hepatic sulphite oxidase activity observed; overall sulphide oxidation capacity was also reduced. Molybdate but not tungstate caused increases in the total plasma copper of guinea pigs but in particular the appearance of a new TCA-insoluble fraction. The effect was increased by the addition of 500 ppm sulphur as sulphide, to the molybdate diet whereas the addition of 500 ppm S as sulphate was ineffective. 100 ppm Mo was less effective as thiomolybdate (MoS4=) than as molybdate. The significance of these results in relation to the role of sulphide in the Cu-Mo-S interaction is discussed.     

Measurement of superoxide dismutase, diamine oxidase and caeruloplasmin oxidase in the blood of thoroughbreds

Methods were developed for the measurement of superoxide dismutase (SOD), diamine oxidase (DAO) and caeruloplasmin oxidase in the blood of thoroughbred horses. These enzymes were measured in 178 normal thoroughbreds stabled throughout the United Kingdom. The relationships between the activities of SOD, DAO and caeruloplasmin oxidase and the blood concentrations of their associated trace metals (copper, zinc and manganese) were studied in 52 of the thoroughbreds. Trace metals were measured by electrothermal atomic absorption spectrophotometry. No relationships were found between the activities of erythrocyte SOD and serum/whole blood copper, zinc and manganese, or serum DAO and serum copper or zinc concentrations. Caeruloplasmin oxidase in equine blood was found to be correlated to serum copper concentration, r = 0.695 (P less than 0.001) over the normal range. Samples from thoroughbreds with trace metal deficiency or toxicity were not available for study. The observed normal ranges for the activity of these enzymes are as follows: SOD: 50 to 200 units per ml whole blood between 5 and 95 percentiles; DAO: 0.1 to 28.5 units per litre (means = 14.8, SD 7.1) and caeruloplasmin oxidase; 11.6 to 35.8 units per ml (means = 23.7, SD 6.0). For numerical simplicity, the activity of DAO is given in units per litre, compared to units per ml for caeruloplasmin oxidase and SOD.     

Use of erythrocyte copper:zinc superoxide dismutase activity and hair or fleece copper concentrations in the diagnosis of hypocuprosis in ruminants

Changes in the activity of superoxide dismutase, a copper-containing enzyme in erythrocytes (ESOD), and changes in copper in hair or fleece were compared with the changes in plasma copper during copper depletion and repletion in steers, lambs and ewes. During depletion the decline in ESOD began later than the decline in plasma copper: the lag varied from 0 to 80 days in individual steers and it was least evident when young rapidly growing lambs were subjected to severe depletion. ESOD activity eventually declined at only one-third to one-seventh of the rate shown by plasma copper, the difference being particularly marked in severely depleted lambs. Repletion of ESOD began after repletion in plasma copper and continued more slowly particularly in growing animals. Hair and fleece copper also responded relatively slowly to depletion and repletion. Low values of ESOD and hair or fleece copper may have diagnostic significance, indicating a more prolonged or intense deficiency of copper and a higher probability of clinical and production responses to copper therapy than low plasma or liver copper values.     

Contrasting effects of nitrofurans on plasma corticosterone in chickens following administration as a bolus or diet additive

Administration of furazolidone as a bolus dose (8-500 mg/kg), produced a decrease in plasma corticosterone in chickens. In contrast, addition of furazolidone or furaltadone to the diet (0.04% or above, 10 days), increased plasma corticosterone. Pre-treatment with a 200-mg/kg bolus of furazolidone or furaltadone did not affect pentobarbitone anaesthesia time in the birds. In chickens pre-treated with a nitrofuran in the diet, however, pentobarbitone anaesthesia time was significantly less than that in controls. Furaltadone in the diet, produced significant increases in the amount of cytochrome P-450 and the activity of aniline hydroxylase in the liver microsomes. It is suggested that nitrofurans given in the diet stimulated corticosterone biosynthesis in the adrenal glands and induced mixed-function oxidase activity in the liver. Nitrofurans given as a bolus did not produce these effects. Furazolidone (200 mg/kg) produced severe anorexia, which lasted 2 days in T-line birds. The anorexia seemed to be associated with tissue damage in the birds rather than the ensuing adrenal cortical insufficiency.     

补铜对奶牛血浆铜含量和铜蓝蛋白活性及其相关性的影响

本试验选取了大庆地区某奶牛群11头泌乳奶牛，检测补铜前后奶牛血浆铜含量和铜蓝蛋白(CP)活性的变化，同时分析了两者相关性。结果表明：补铜前血浆铜在10.93 μmol/L以下者占被检牛数的54.55%（6/11），其血浆CP活性降低，两者呈现中度相关性（R2=0.53）；补铜后血浆铜含量与CP活性均极显著增高（P<0.01），血浆铜含量达到正常范围，两者呈现低相关性（R2=0.12）。〖JP〗结论：该奶牛场泌乳奶牛群中存在低铜血症病牛，血浆铜蓝蛋白可以作为奶牛低铜血症的一项检测指标。     

Injection of sheep with organic compounds of copper

Subcutaneous injections of up to 6 mg copper per kg body-weight as copper methionate (A) produced no deleterious effects in ewes. When copper was given in the form of copper calcium EDTA (B), deaths occurred within 48 hours of the subcutaneous injection of 3 or 4 mg copper per kg body-weight. There was centrilobular necrosis of the liver, necrosis of kidney tubules and excess fluid in the pleural and peritoneal cavities accompanied by the release of liver specific enzymes and a rise in the concentration of urea and creatinine in serum. The injection of 2 mg copper per kg body-weight produced a liver lesion at 48 hours with similar serum changes but recovery took place. Liver and kidney lesions occurred when the copper was in the form of diethylamine copper oxyquinoline sulphonate (C). In this case deaths occurred after the injection of 4, 3 or 2 mg copper per kg body-weight and liver damage followed by recovery was caused by 1 mg copper per kg body-weight. All compounds produced an increase in copper oxidase activity and in the concentration of copper in serum and whole blood. Compound A produced a slow increase in plasma copper concentration to a plateau between 5 and 10 days. Compounds B and C produced a rapid initial increase in the first few hours. The concentration fell rapidly for 12 or 24 hours and then more gradually. It is suggested that rapidity of absorption and transfer of copper to the liver and kidneys may be responsible for the toxic effects of compounds B and C.     

Sexual difference in ascorbic acid synthesis, tissue ascorbic acid and plasma total antioxidant capacity in mature chickens

1. An experiment was conducted with commercial White Leghorn type chickens to determine the effect of gender on tissue ascorbic acid concentration, antioxidant capacity and ascorbic acid synthesis. 2. Birds reared and maintained on litter were given a standard layer diet, without supplemental ascorbic acid, from 18 weeks of age. Tissue ascorbic acid concentration, plasma total antioxidant capacity and renal L-gulonolactone oxidase activity were measured at 30 weeks of age. 3. Females and males differed in ascorbic acid synthesis, as measured by renal L-gulonolactone oxidase activity, and tissue ascorbic acid concentration. 4. Plasma total antioxidant capacity and adrenal, gonadal, plasma and pituitary ascorbic acid concentrations were significantly higher in males, whereas ascorbic acid synthesis and splenic and thymic ascorbic acid concentrations were significantly higher in females. 5. L-Gulonolactone oxidase activity was not detected in the comb of cockerels.     

Changes in the ultrastructure of cardiac muscle in steers deprived of copper.

Young Friesian steers were fed for nine months on copper supplemented or depleted diets. Immediately after they had been killed, tissues were taken from the myocardium for examination by electron microscopy. Copper deficiency was associated with extensive changes in myofibrillar and mitochondrial morphology and distribution. It is suggested that the ultra-structural changes in the myocardium were specifically attributable to copper deficiency, the most likely underlying biochemical defect being lost of cytochrome oxidase activity.     

桑属几个种及品种细胞色素氧化酶同工酶的研究

用聚丙烯酰胺凝胶电泳的方法,对四川桑属植物的6个种、两个品种及两个引进种的细胞色素氧化酶同工酶进行了研究。结果:(1)桑属内有共同的特征酶带;(2)桑属内各种、品种间的酶谱有显著差异,每个种都有不同的特征酶谱;(3)用组平均聚类法研究同工酶分类,在桑属植物分类上有一定的适用价值;(4)上述研究结果与过氧化物酶同工酶的排序法分类比较,除华桑有差异外,其它桑种、品种均表现规律性的一致倾向,为桑属植物分类提供了新的依据。     

绵羊实验性钼中毒研究——Ⅲ.硫酸盐对体内钼、铜、铁状态的影响

日摄食钼250ppm[15mg/(kg·d)]的成年绵羊,加喂硫酸钠7g/d能显著减少其血浆、被毛及组织中的钼蓄积,显著增加脾、肝、肾等组织的铁含量;而单纯摄入高钼则对体内铁状态无显著影响。硫酸盐对摄食高钼绵羊体内铜状态的影响与单纯投钼羊类似,表现为肝、肾等组织铜、血浆总铜以及铜蓝蛋白氧化酶(CP)活性增加;对血浆铜的组分构成具有显著影响,表现为降低三氯醋酸(TCA)不溶铜而升高TCA可溶铜。在实验期间,加喂硫酸盐并未引起钼诱导的铜缺乏。     

Effects of phenobarbital treatment on 3-methylindole toxicosis in ponies

To study the role of cytochrome P-450-dependent mixed function oxidase reactions in equine 3-methylindole (3MI) toxicosis, ponies were given 20 mg of phenobarbital/kg of body weight at 72, 60, 48, 36, and 24 hours before 100 mg of oral 3MI/kg to induce cytochrome P-450 or no treatment (controls). Maximal 3MI plasma concentration was decreased and clearance was faster in phenobarbital-treated ponies. Plasma 3MI was still detectable 12 and 36 hours after dosing in phenobarbital-treated and control ponies, respectively. Phenobarbital treatment induced a distribution phase with transition from a 1-compartment to a 2-compartment extravascular model. Bronchiolitis occurred in all ponies 72 hours after 3MI, but was more severe in those treated with phenobarbital. Appearance of a distribution phase, increased total body clearance, and more severe bronchiolitis in phenobarbital-treated ponies indicated that mixed function oxidases are involved in metabolism and conversion of 3MI to a toxic metabolite.     

Serum copper and ceruloplasmin activity at the early growing stage in foals.

Serum concentrations of copper (Cu), zinc (Zn), manganese (Mn), calcium (Ca) and inorganic phosphorus (P), as well as antigenic ceruloplasmin (Cp) and oxidase activity as a functional index for copper metabolism, were measured in 10 foals (5 males and 5 females) and their dams. Samples were harvested from the foals within 1 wk after birth and monthly from 1 to 17 mo of age. Samples were collected from their dams in the perinatal period (monthly from 2 mo before delivery to 5 mo postpartum). Serum oxidase activity, antigenic Cp and Cu in foals were extremely low at 1 wk. Serum Cp had the lowest value of 17.0 +/- 8.0 (mean +/- SD) mg/dL within the 1st wk, then increased rapidly up to 43.7 +/- 5.8 mg/dL at 1 mo, and maintained this level until the 17th mo. Serum Zn in foals had the highest value of 73.2 +/- 13.1 micrograms/dL within 1 wk, then decreased to 38.3 +/- 5.9 micrograms/dL by 17 mo. Serum Mn, Ca and P in mares were almost stable and within established reference ranges for our laboratory in the perinatal period, and these values in foals were also in the normal range. Even on appropriate feeding, serum Cu, Cp and oxidase activity were quite low a few weeks after birth, while a higher proportion of Cp-binding copper was found in the foals. This might be caused by the limited synthesis of ceruloplasmin in this period. These data suggest that newborn foals are in a critical situation of marginal copper status in the early stage of growth.