The Concentrations of Molybdenum and Zinc in Liver in Relation to Copper Accumulation in Normal and Copper Poisoned Sheep

The concentrations of copper, molybdenum and zinc were measured in the liver of normal grazing sheep and lambs from Eastern Norway, and in sheep dead of chronic copper poisoning. The following mean values were found: Normal sheep: 173 ± 130 μg Gu/g wet weight, 1.0 ±0.3 μg Mo/g, and 49 ± 10 μg Zn/g; lambs: 129 ± 59 μg Gu/g, 0.9 ± 0.3 μg Mo/g, and 46 ±9 μg Zn/g; sheep dead of copper poisoning: 429 ± 249 μg Gu/g, 0.4 ± 0.1 μg Mo/g, and 43 ± 2d μg Zn/g. Sheep with low liver copper (Gu < 10 μg/g) were also analyzed for molybdenum and zinc, with the following results: 1.0 ± 0.2 μg Mo/g, and 45 ± 8 μg Zn/g wet weight. The differences in liver copper between all the groups, and the differences in molybdenum concentrations between the normal sheep and the lambs and between the normal sheep and the poisoned sheep were significant (P < 0.001). No significant correlations between liver copper/liver molybdenum or liver copper/liver zinc were detected.     

Copper, zinc and molybdenum in goat liver.

The concentrations of copper, zinc and molybdenum were measured in the liver of goats from western and south-eastern Norway. The mean copper concentrations in the liver of goats from these two districts were 23±19 µg Cu/g and 59±31 µg Cu/g wet weight, respectively. As for zinc and molybdenum, no difference was found between the two groups of animals. No correlations were detected between copper and zinc, zinc and molybdenum, or copper and molybdenum. The copper levels in Norwegian goat liver are considerably lower than in sheep liver, and the ranges are significantly more narrow. The concentrations of molybdenum in goat liver are at the same levels as in sheep and swine, while the levels of zinc are somewhat lower.     

Copper,Zinc and Molybdenum in Livers of Norwegian Cattle At Slaughter

The concentrations of copper, zinc and molybdenum were measured in samples of cattle liver from 10 slaughter-houses in Norway. A total of 335 samples were analysed. A clear accumulation of copper with age was found, the average copper level in the younger animals (≦ 3 years, n = 194) being 30 µg Cu/g liver wet weight, and in the older ones (> 3 years, n = 141) 59 µg Gu/g. The range in the copper values found was considerable, though significant differences between some of the districts were recorded. Copper concentrations were classified as low (≦10 µg Gu/g) in 9.6 % of the samples. Zinc showed no accumulation with age, nor were there any differences in zinc levels found in animals from different districts, the average level being 32 µg Zn/g liver wet weight. The picture was the same for molybdenum, no differences between age groups or districts being found. The average level was 1.0 µg Mo/g liver. There was no significant correlation between levels of copper, zinc or molybdenum.The supply of copper and zinc to cattle in Norway seems close to sufficient, but copper- and zinc-fortified mineral supplementation of cattle feed is still to be recommended. There seems to be no need for molybdenum supplementation in cattle.     

Evaluation of a commercially available molybdate formulation and zinc oxide boluses in preventing hepatic copper accumulation and thus enzootic icterus in sheep

The efficacy of a molybdate formulation and a zinc oxide bolus as prophylactic agents for enzootic icterus was evaluated in sheep. Before copper loading, liver biopsies were performed on 12 male, 6-month-old, Mutton Merino sheep to determine hepatic copper (Cu) and zinc (Zn) concentrations. The animals were restrictively randomised according to liver copper concentrations to 3 treatment groups (n = 4) to achieve similar mean liver copper concentrations per group. All sheep received 4 ml/kg of a 0.5 % aqueous solution of CuSO4 5H2O intraruminally 7 days per week for 10 weeks. On Day 0 the sheep in the Mo-group were injected subcutaneously with 42 mg molybdenum (Mo) contained in a commercial molybdate formulation. The animals in the Zn-group each received a zinc oxide bolus, containing 43 g zinc oxide, via a rumen cannula. Treatment was repeated on Day 42. Four animals served as untreated controls. Urinary copper excretion, plasma copper concentration, haematocrit and glutamate dehydrogenase (GLDH) activity were determined throughout the trial. The animals were sacrificed after 10 weeks and liver samples were submitted for histopathological examination. Liver and kidney copper and zinc concentrations were determined. Neither the molybdate treatment nor the zinc oxide boluses prevented hepatic copper accumulation. The urinary copper excretion, plasma copper concentration, haematocrit and GLDH activity were not significantly different (P > 0.05) from the controls.     

The Concentrations of Copper,Zinc and Molyrdenum in Swine Liver and the Relationship to the Distrirution of Solurle Copper- and Zinc-Rinding Proteins

The concentrations of copper, zinc and molybdenum were measured in liver samples from 21 normal slaughter pigs (average age about 6 months) and in 36 sows (average age about 2 years). The following mean values were found: Slaughter pigs: 15 ± 8 µg Cu/g, 45 ± 7 μg Zm/g and 1.0 ± 0.2 μg Mo/g wet weight; sows: 46 ± 70 μg Cu/g, 70 ± 26 μg Zn/g and 1.3 ± 0.3 μg Mo/g wet weight. The concentrations of all 3 elements were significantly higher in the sows than in the young pigs. There was no correlation between the concentrations of copper, zinc or molybdenum. The recorded copper levels in the slaughter pigs were in accordance with the levels of non-supplemented pigs given in the literature. The soluble hepatic copper- and zinc-binding proteins were separated into 3 different fractions by gel filtration. With increasing copper and zinc levels in the liver, a higher relative amount of these elements were found in the low molecular weight fraction.     

Trace elements in grass, cattle liver and sheep liver from districts surrounding Karatina, Kenya. II. Copper, molybdenum, zinc and sulphur

Thirty-one samples of pasture grass from districts surrounding Karatina, Kenya were analysed for contents of copper, molybdenum, zinc and sulphur. The following mean values and standard deviations were found: Copper: 8.2 +/- 5.0 mg/kg DM; molybdenum: 1.4 +/- 2.6 mg/kg DM; zinc: 33 +/- 10 mg/kg DM and sulphur: 0.17 +/- 0.06% DM. Mean value of the ratio between copper and molybdenum was 13 +/- 11. Samples of cattle liver (n = 96) and sheep liver (n = 93) were analysed for copper and zinc with the following results: Cattle liver: 21 +/- 16 mg Cu/kg WW and 37 +/- 11 mg Zn/kg WW. Sheep liver: 59 +/- 37 mg Cu/kg WW and 30 +/- 6.4 mg Zn/kg WW. It is concluded that subclinical copper deficiency may occur in cattle in the districts included in the investigation. The copper status of sheep seems to be adequate. The levels of zinc may indicate a marginal intake of this element in sheep.     

青海省部分地区藏羊肝脏六种微量元素水平的测定

对来自青海省海西州和海北州部分地区的37份藏羊肝脏样品中的铜、铁、钴、硒、钼和硒等六种微量元素水平进行了检测,结果显示,肝铜含量在34.45±7.37~168.62±57.85之间,处于铜缺乏状态;肝硒水平在0.0169±0.0026~0.0323±0.0033之间,各地均处于极度缺硒状态;肝钴水平在正常范围之内;肝铁平均水平为3572.21±1155.56μg/g,严重偏高;肝钼的水平略低于正常值;肝锌水平略高于正常水平。     

Histochemigal Detection of Copper in the Liver from Sheep Chronically Poisoned by Copper

Continued ingestion of copper in excess of the nutritional requirement leads in all animals to its passive accumulation within the tissues, especially the liver. Up to certain levels varying greatly with the species high concentrations of copper in the liver appear to impose no physiological hardship on the animal. Above these levels there may occur a catastrophic liberation of a high proportion of the copper into the blood stream resulting in extensive hemolysis and jaundice usually followed by death. Sheep are more prone to the hemolytic crisis of chronic copper poisoning than other animal species. The hemolytic crisis of chronic copper poisoning is associated with centrolobular necrosis of the liver. The copper concentration in the liver is usually 300 p.p.m. or more (wet tissue) compared with about 50 p.p.m. in normal sheep (Simesen & Møller 1969).     

X-ray microanalysis of liver and kidney in copper loaded sheep with and without thiomolybdate administration

The distribution of elements in the cells of the liver and kidney of normal sheep, of sheep chronically intoxicated with copper and in sheep given copper and thiomolybdate was studied by atomic absorption spectrophotometry, electron microscopy and electronprobe X-ray microanalysis. Copper concentrations were increased in the liver and kidney during the pre-haemolytic and haemolytic stages of the disease. In addition iron concentrations were markedly increased in the kidney during haemolysis. Copper and molybdenum concentrations were increased in the kidney of sheep receiving copper and thiomolybdate or thiomolybdate alone. By electronprobe X-ray microanalysis at the pre-haemolytic and haemolytic stages, copper together with small amounts of calcium and chromium were found in lysosomes of liver cells. In addition, in animals killed during haemolysis, high concentrations of iron were found in apical lysosomes of kidney proximal tubule cells. Copper and molybdenum were found in apical lysosomes of kidney tubule cells in animals given copper and thiomolybdate or thiomolybdate alone. The accumulation of copper with molybdenum and sulphur in the lysosomes of thiomolybdate treated copper intoxicated animals was demonstrated for the first time and may indicate the sequestration within lysosomes of a copper-molybdenum-protein complex.     

Copper poisoning in a flock of sheep. Copper excretion patterns after treatment with molybdenum and sulfur or penicillamine

During an outbreak of chronic copper poisoning, fecal and urinary copper excretion were measured following treatment with molybdenum and sulfur supplementation of the feed (0.1 g ammonium molybdate plus 1 g sodium sulfate/sheep/day) or oral penicillamine (50 mg/kg bodyweight/day) using rams in metabolism cages. Serum glutamic-oxaloacetic transaminase activities and liver levels of molybdenum and copper in sheep that died were also monitored. Within four days of starting molybdenum and sulfur supplementation a highly significant increase in fecal copper excretion was evident and the increase persisted throughout the monitoring period (five weeks — general treatment of the flock continued for another three weeks). There was no effect of the molybdenum and sulfur supplementation on urinary excretion of copper. The molybdenum and sulfur supplementation was very effective, resulting in a rapid marked decrease in mortality. Oral penicillamine treatment induced cupruresis but did not affect fecal copper excretion. The results indicated that, while the cost of penicillamine may be a limiting factor for general treatment of a flock, it may be the drug of choice for the therapy of valuable breeding animals because cupruresis may be accurately and individually controlled. Serum glutamicoxaloacetic transaminase activities were a valuable aid in diagnosing chronic copper toxicosis as well as for monitoring recovery. High initial liver copper levels were gradually reduced following molybdenum and sulfur treatment. However, at the end of the study the liver copper levels of dead sheep varied within wide limits and there were still some sheep with high liver copper levels.     

Studies on the Haematology and Trace Element Status of Adult Bactrian Camels (Camelus bactrianus) in China

Reference values were established for some haematological and serum biochemical constituents in Bactrian camels (Camelus bactrianus) in China. The contents of seven trace elements in the blood, hair, liver, kidney, heart, spleen, lung, cerebrum, cerebellum, rib, ovary, pancreas and gluteal muscle of Bactrian camels were also measured. Some of these values are reported for the first time for Bactrian camels in China. Most haematological and serum biochemical values were similar to those of cattle, yaks, sheep and dromedary camels, but the mean serum albumin concentration and the albumim/globulin ratio were significantly higher than those in other ruminants and the mean thyroxine concentration was half that in dromedary camels. The liver contained the highest concentrations of copper, zinc, manganese and iron, and the renal cortex contained the highest concentration of selenium. The concentrations of selenium, cobalt, zinc, manganese and molybdenum in the tissues were within the reference ranges for other ruminants, but the mean iron and copper concentrations in the liver were significantly higher than those in other ruminants.     

戈壁草原蒙古羊春季羔羊铜缺乏症的研究

[目的] 确定内蒙古苏尼特左旗北部地区春季发生的戈壁草原蒙古羊羔羊发病、死亡现象的主要原因。[方法] 选取3户羔羊发病情况最为严重的牧户采样,每户各采集5只发病羔羊的血液样本、放牧草场的饮用水样本,以及羔羊所处环境中3种主要牧草（红砂、珍珠猪毛菜和盐爪爪）样本和土壤样本,同时采集当地种羊场15只相同品种健康羔羊的样本作为对照,测定上述样本中的10种矿物质元素（钙、磷、铁、锰、铜、锌、镁、钼、硒、钴）的含量。[结果] 发病羔羊血液中铜和镁的含量显著（P<0.05）低于同品种健康羔羊,钼的含量显著（P<0.05）高于同品种健康羔羊,其他矿物质元素的含量差异不显著（P>0.05）。发病羔羊所处环境中饮用水的镁和钙含量较高。发病羔羊所处环境中红砂的钼、铁、锰和硒含量均高于文献报道值,其他矿物质元素含量均处于正常范围;盐爪爪的钼、铁、锰、硒、镁和钴的含量高于文献报道值,其他矿物质元素含量均处于正常范围;珍珠猪毛菜、红砂和盐爪爪的铜钼比分别为15.96∶1、0.23∶1和2.34∶1。发病羔羊所处环境中土壤样本的铜、铁、锰、硒、钴含量均低于文献报道值,镁、钙和磷含量均高于文献报道值,钼和锌含量在正常范围内。[结论] 该地区戈壁草原蒙古羊羔羊发病、死亡的原因主要是放牧羊摄入铜不足、钼超标、铁和锰等矿物质元素摄入量过多,导致羔羊对铜的吸收发生障碍,最终引起继发性铜缺乏症。     

Fractionation of Soluble Copper- and Zinc-Binding Proteins From Cattle Liver

The distribution of copper and zinc among soluble proteins in liver from normal slaughter cattle was examined after gel filtration of the proteins. Gopper- and zinc-binding proteins were mainly separated into three fractions. Varying amounts of zinc were eluted in a fourth fraction of molecular weight less than 2,000. A clear relationship was noted between the amount of copper bound to the low molecular weight fraction (m.w. ~ 10,000) and the total liver zinc concentration. The high molecular weight protein fraction (m.w. > 65,000) dominated in liver with zinc concentrations below 40 µg/g wet weight and total copper concentrations from 16 to 240 µg/g, while in liver with zinc concentrations above 40 µg/g and copper concentrations ranging from 20 to 107 µg/g, the low molecular weight metallothionein-like fraction dominated.     

Effect of zinc and molybdenum supplementation of the feed concentrate on the storage of copper in the liver of lambs

Summary During a 98-day trial, eight groups of eight male weanling Texel x Friesian milk sheep lambs were fed the same hay and concentrates, the latter differing only in the contents of Cu, Zn, and Mo. At high (± 30 ppm) and moderately low (± 15 ppm) copper levels in the concentrates, additions of Zn (respectively 0, 150, and 340 ppm) and Mo (0 and 16 ppm) to the concentrates were studied with regard to effect on the accumulation of copper in the liver. At the higher copper level, the addition of 150 or 340ppm Zn resulted in a significant decrease of the Cu accumulation in the liver, while at a moderately low copper level the effect of these additions on the Cu accumulation was only slight. The two Zn doses given appeared to be equally effective in reducing the Cu accumulation. Additions of zinc did not increase liver and kidney Zn. The addition of Mo. resulted in higher levels of Mo in liver and kidney. At either copper level the addition of 16 ppm Mo was more effective in reducing the Cu accumulation in the liver than the addition of 150 or 340 ppm Zn. The additions of Zn and Mo, had no visible influence on the health of the lambs. This agrees with the finding that the blood parameters (determination of haemoglobin, haematocryt, protein, copper, zinc, enzymes) and the growth of the lambs were in the normal range. These results suggest that zinc may be a useful alternative to the somewhat toxic molybdenum as an additive to concentrates for sheep in preventing Cu intoxication.     

The effect of lupinosis on liver copper, selenium and zinc concentrations in merino sheep

Merino wether weaners were exposed to toxic lupin stubbles for periods of one, two and six to nine weeks, and the effect on their liver copper, selenium and zinc concentrations studied. After the one week period there was a slight decrease in liver copper and selenium concentrations in sheep severely affected with lupinosis. This was attributed to loss of these elements from necrotic cells, and greatly increased quantities of fat in the liver. In the same period liver zinc concentrations declined and were negatively correlated with the degree of liver injury. After two weeks or more of exposure to toxic lupins a positive correlation existed between both the liver copper and selenium concentrations, and the degree of liver injury. Furthermore, total liver copper and selenium levels were also positively correlated with the degree of liver injury. Liver zinc concentrations were negatively correlated with the degree of liver injury. It is suggested that when the liver is under the influence of the toxins causing lupinosis for more than two weeks, it stores copper and selenium, and loses zinc.     

Fractionation of Soluble Molybdenum-Binding Proteins from Liver,Kidney, Plasma and Erythrocytes from Sheep Supplemented with Molybdenum

Five ewes were given a daily dose of 1.4 mg Mo/kg body weight as ammonium molybdate for 21 to 55 days. Plasma molybdenum concentrations increased considerably following the dosage, but no systematic changes were seen in the plasma copper concentrations. About 80 % of the molybdenum present in blood was found in plasma. With gel filtration it was shown that practically all of the molybdenum present in plasma and red cell haemolyzates was found in a fraction with molecular weight less than 1,500. Soluble proteins from liver and kidney cortex were separated into one high molecular weight fraction with molecular weight above 200,000 and a non-protein fraction with molecular weight less than 1,500. Significant correlations were found between the amounts of molybdenum present in the high molecular weight fraction and the molybdenum concentrations in liver and kidney.     

A study of high lamb liver copper concentrations on some farms in Otago and Southland

Lamb copper status is commonly assessed by measuring copper concentrations in four liver samples collected from lines of lambs sent to meat slaughtering premises. High liver copper concentrations were found in lambs examined in April and May. Five farms with two or more lamb liver copper concentrations greater than 3000 𝛍mol/kg on a wet matter basis and two farms with adequate copper concentrations but less than 3000 𝛍mollkg in the autumn of 1992 were selected for a more detailed investigation into the factors affecting their lamb copper status in 1993. High liver copper status was associated with low pasture molybdenum, grazing paddocks recently topdressed with copper sulphate, supplementing with mineralised drenches and copperised salt licks, and the high copper content in chicory. Lamb liver copper concentrations were significantly (pp<0.05) higher in the autumn than in the summer on two of three farms.     

Interaction between copper and zinc in sheep parenterally treated with zinc oxide

Changes in copper and zinc concentrations in the blood serum of sheep were studied in clinical conditions for sixty days in relation to the intramuscular application of zinc oxide at a dose of 10 mg Zn per kg liveweight; zinc oxide was administered in a developmental preparation. Serum concentrations of zinc started increasing significantly the eighth day after administration (p less than 0.01), the maximum values were found out on the twelfth day (21.19 +/- 3.89 mumol X l-1). Significantly higher concentrations of zinc in the blood serum of test sheep were observed as soon as on the eighth day (p less than 0.01), in comparison with the untreated group, and they persisted till the end of investigation (p less than 0.01). Simultaneously with the changes in serum zinc concentrations, copper metabolism was influenced when the statistically higher values of copper concentrations in the blood serum of the test group were recorded the eighth and twelfth days (p less than 0.01), in comparison with the control animals. In the remaining period, serum copper concentrations did not show any mathematical dependence and the equalization to the original values in the treated group occurred at the end of the experiment. The above-mentioned results from the sheep treated with zinc oxide refer to a possibility of influencing mutually the copper and zinc metabolism also parenterally; this could be utilized during the treatment of secondary deficiencies, or toxicosis.     

Renal lesions and tissue concentrations of zinc, copper, iron and manganese in experimentally zinc intoxicated sheep

Renal lesions were produced in three out of 10 sheep dosed with large quantities of zinc for two to 14 weeks. Administration of the zinc resulted in significant increases in the concentrations of zinc in the kidneys, liver and pancreas, and of copper and iron in the kidneys and pancreas, and a significant decrease in the concentration of manganese in the pancreas. The sheep with kidney damage had increased plasma creatinine concentrations and significantly higher mean concentrations of zinc in their livers and pancreases, and of copper and iron in their kidneys, than the sheep with no kidney damage.     

The Distribution of Soluble Copper- and Zinc-Binding Proteins in Liver and Kidney of Chronic Copper-Poisoned Goats

The distribution of copper and zinc among the soluble proteins in the liver and kidney from chronic copper-poisoned goats was examined after gel filtration of the proteins. The concentrations of copper in the liver and kidney cortex from five experimentally copper-poisoned goats were: 550–810 µg/g liver and 190–420 µg/g kidney cortex (wet weight). In general the copper-binding proteins from, both the liver and kidney samples were separated into two different fractions with approximate molecular weights (m.w.) of > 65,000 and 10,000, respectively. From the liver samples, varying amounts of copper were eluted in a fourth fraction with m.w. < 2,000. In the majority of kidney samples the dominating copper-binding protein fraction was the high molecular weight fraction. Absolute amounts of copper recovered in the metallothionein-like protein fraction were nearly the samt for all samples investigated. The distribution of zinc-binding proteins in both liver and kidney samples was nearly the same. The high molecular weight fraction dominated, and no zinc was bound to metallothionein-like proteins.