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 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.     

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.     

Concentration of Selenium in Liver in Relation to Copper Level in Normal and Copper-Poisoned Sheep

Copper absorption, liver accumulation and development of copper toxicosis in sheep are influenced by a variety of other elements, in particular molybdenum, sulphur and zinc (Underwood 1977). In a previous study on liver concentrations of copper, molybdenum and zinc in normal and copper-poisoned sheep, no direct correlation was found between the concentrations of the three metals, but molybdenum was significantly lower in the livers from sheep dead from chronic copper poisoning than in normal animals (Frøslie & Norheim 1976).     

Effects of Daily Oral Administration of Copper to Goats

Three goats were dosed orally with a 0.2 % aqueous copper sulphate solution. The dosing was 20 mg copper sulphate/kg body weight twice a day for 56 to 113 days. One of the goats accumulated substantial amounts of copper in the liver and developed two haemolytic crises. The two other goats showed only increased liver copper concentrations before they were killed. The results indicate that the goats were susceptible in varying degrees to repeated oral copper dosing, and that two of the goats were significantly less susceptible to copper than sheep. The goat that turned into a haemolytic crisis showed changes similar to those seen in sheep as far as blood and plasma parameters are concerned. The gross and histological lesions were also mainly of the same type as described in sheep. The hepatic lesions found in the goat differed to some degree from those found in sheep as the necroses were more distinctly located to the centrilobular area, and as the iron pigments were mainly located in phagocytes in the hepatic sinusóides.     

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.     

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 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.     

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.     

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。     

Dietary Management of Labrador Retrievers with Subclinical Hepatic Copper Accumulation

Background

Genetic and environmental factors, including dietary copper intake, contribute to the pathogenesis of copper‐associated hepatitis in Labrador retrievers. Clinical disease is preceded by a subclinical phase in which copper accumulates in the liver.

Objective

To investigate the effect of a low‐copper, high‐zinc diet on hepatic copper concentration in Labrador retrievers with increased hepatic copper concentrations.

Animals

Twenty‐eight clinically healthy, client‐owned Labrador retrievers with a mean hepatic copper concentration of 919 ± 477 mg/kg dry weight liver (dwl) that were related to dogs previously diagnosed with clinical copper‐associated hepatitis.

Methods

Clinical trial in which dogs were fed a diet containing 1.3 ± 0.3 mg copper/Mcal and 64.3 ± 5.9 mg zinc/Mcal. Hepatic copper concentrations were determined in liver biopsy samples approximately every 6 months. Logistic regression was performed to investigate effects of sex, age, initial hepatic copper concentration and pedigree on the ability to normalize hepatic copper concentrations.

Results

In responders (15/28 dogs), hepatic copper concentrations decreased from a mean of 710 ± 216 mg/kg dwl copper to 343 ± 70 mg/kg dwl hepatic copper after a median of 7.1 months (range, 5.5–21.4 months). Dogs from a severely affected pedigree were at increased risk for inability to have their hepatic copper concentrations normalized with dietary treatment.

Conclusions and Clinical Importance

Feeding a low‐copper, high‐zinc diet resulted in a decrease in hepatic copper concentrations in a subset of clinically normal Labrador retrievers with previous hepatic copper accumulation. A positive response to diet may be influenced by genetic background. Determination of clinical benefit requires further study.     

Copper,iron, zinc and tannin concentrations throughout the digestive tract of tropical goats and sheep fed a high-fibre tannin-rich diet

The dry season in tropical regions urges livestock to feed on nutritionally very poor diets. It has not been explored how tropical sheep—assumed grazers—and tropical goats—intermediate browsers—cope with a high-fibre tannin-rich diet. This study was designed to determine the effects of a high-fibre and tannin-rich diet on the flow of important microminerals iron (Fe), zinc (Zn) and copper (Cu) throughout the digestive tract of tropical sheep and goats. The feeding trial was set up with twelve adult male animals, six sheep with mean body weight (BW) of 30.3 ± 1.6 kg and six goats with mean BW of 26.4 ± 2.2 kg. The feed consisted of 36% leaves of Millettia ferruginea, 61% hay and 3% concentrate and was offered at 3% of BW (all on dry matter (DM) basis). The total faecal collection was carried out for 7 consecutive days. At the end of the experimental period, the animals were slaughtered to collect liver and digesta samples from the gastrointestinal tract. Feed, digesta and faecal samples underwent analysis of Fe, Zn, and Cu and total tannins (TT). Goats had significantly higher reticulum Cu concentrations expressed on DM as compared to sheep. Faecal Cu concentrations were higher for goats compared to sheep. Reticulum and colon digesta Zn levels were higher in goats than sheep. Abomasum and colon Fe levels were higher in sheep than goats when expressed on DM. These results suggest differences in feed intake, micromineral absorption, secretion and excretion between sheep and goats, pointing to a divergent mineral metabolism as an adaptation to the challenge of a dry season diet having very low nutritive value.     

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.     

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。     

An evaluation of the effect of clotting on the recovery of copper from caprine blood

In the goat, diagnosis of copper (Cu) deficiency is often based on measurement of Cu in serum or plasma. Previous research in cattle and sheep has shown that these values are not interchangeable, but data for goats have not been published. Paired serum and heparinised plasma samples taken from 119 goats in eight herds were tested for Cu concentration. Plasma and serum Cu were significantly correlated (r=0.95). On average serum Cu was 3.5 μmol/L lower than plasma Cu, but this difference was related to Cu status (r=0.45). Mean serum Cu concentration was 83% of plasma Cu, with the 95% limits of agreement ranging from 66% to 100%. Similar to cattle and sheep, individual variability in Cu loss during clotting is too great for serum Cu to be used as a measure of Cu status in goats.     

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.     

The effects of plastic slatted floor or straw bedding on performance, liver weight and liver copper concentrations in intensively reared lambs

One hundred and eight Texel sired lambs were weaned at 37 days old and allocated three days later to one of two treatments, using two pens per treatment, in order to evaluate lamb hepatic copper (Cu) values. In Treatment 1 (T1) the lambs were confined to pens having recycled plastic slatted flooring, while in Treatment 2 (T2) the lambs were confined to pens having straw bedded floors, with fresh straw added on alternate days. The lambs had ad libitum access to a coarse lamb concentrate. The crude protein (g/kg DM), crude fibre (g/kg DM), total zinc (mg/kg) and background levels of copper (Cu) (mg/kg) and molybdenum (Mo) (mg/kg) of the diet were 17.2, 10.9, 152, 9.7 and 0.92, respectively. The lambs were slaughtered when they reached about 39 kg live weight. Following slaughter, the liver was removed and weighed and the caudal lobe frozen for subsequent Cu analysis. The livers of lambs on the plastic floor treatment had higher liver weights (843 vs. 804 g, S.E.M. 11.0; P < 0.05) and higher hepatic copper (396 vs. 315 mg/kg DM, S.E.M. 18.2; P < 0.05) than those from lambs that were bedded with straw. The Cu accumulation per kg of liver DM per week was estimated to be 8.4 and 13.7 mg for lambs on the straw bedding and plastic slated treatments, respectively. It is concluded that when lambs are housed on plastic slatted flooring, relative to straw bedding, there is a higher accumulation of hepatic copper levels likely to result in a greater risk of Cu toxicity when lambs are reared on all concentrate diets.     

Effect of sex and rearing system on the quality and mineral content of fiber from raeini cashmere goats

The aim of this study was to compare the quality characteristics and mineral content of the fiber from male and female cashmere goats raised under different management systems. Male and female Raeini cashmere goats (<1.5 years of age, n = 48) were selected from flocks raised at a government breeding station or raised commercially under either rural or nomadic conditions. The staple length, cashmere fiber diameter, coefficient of variation for fiber diameter, percentage of cashmere in a fleece, percentage of guard hair in a fleece and cashmere tenacity averaged 4.6 ±0.1 cm, 18.0 ±0.1 μm, 20.9 ± 0.4%, 66.1 ± 1.5%, 33.8 ± 1.5% and 1.8 ± 0.2 gf/tex, respectively. The sulfur, copper and zinc content of the cashmere averaged 2.8 ± 0.1%, 0.00065 ± 0.00002% and 0.01276 ± 0.00025%, respectively. Rearing method significantly affected staple length, coefficient of variation of fiber diameter, cashmere tenacity and copper content. Males had a higher coefficient of variation of fiber diameter and cashmere tenacity than females (P < 0.05).     

Copper toxicosis in a Boer goat

A 1‐year‐old female Boer goat was presented with a 1‐day history of pigmenturia, anorexia, and shivering. Anemia was not present initially, but progressive hemolytic anemia developed subsequently and was characterized by the finding of Heinz bodies in both intact RBCs and in ghost cells and the presence of atypical fusiform RBCs. Plasma biochemical analysis revealed increased activities of aspartate aminotransferase and gamma‐glutamyltransferase, hyperbilirubinemia, and azotemia. Histopathologic examination of a liver biopsy revealed necrosis of individual hepatocytes and intracytoplasmic rhodamine‐positive granules, consistent with copper. Copper concentration in ante‐mortem hepatic tissue was increased, and a diagnosis of copper toxicosis was made. Despite supportive therapy, the goat continued to decline and was euthanized. Necropsy findings included hepatic necrosis and hemoglobinuric nephrosis. Freshly collected specimens of liver and kidney had markedly increased copper concentrations. The mineral composition of the water, grass hay, and goat chow was evaluated, and toxins and significant mineral imbalances were not found. The underlying cause of the hepatic accumulation and subsequent release of copper remains unclear in this goat. Recently, Boer goats have been recognized as being prone to copper toxicosis and may be more susceptible than other breeds; similar to sheep, Boer goats may experience a hemolytic crisis secondary to copper toxicosis.     

Luteal lifespan and fertility after estrus synchronization in goats

The present experiment aims to examine the efficiency of estrus synchronization using progesterone and equine chorionic gonadotrophin (eCG) and to look at luteal function. During the non-breeding and breeding season, 5 adult female Korean native goats were injected intramuscularly with 2.5 ml of physiological saline as the control. A progesterone impregnated intravaginal sponge was then kept in the same goats for 10 days followed, after a week, by an intramuscular injection of 500 IU eCG. Five adult female Nubian goats were mated with a fertile buck during the non-breeding season. During the non-breeding season 2 of the 5 goats showed a normal estrous cycle (ranging from 18 to 21 days) and 3 a short estrous cycle (ranging from 3 to 6 days). During the breeding season the equivalent figures were 1 and 2. The major axes of the corpus luteum (CL) were measured by means of calipers built into the ultrasonography system, and the concentrations of plasma progesterone (P₄) were determined by double antibody radioimmunoassay. The mean major axes of the CL in goats showing the short cycle (6.1 ± 0.5 mm) was significantly smaller than in those showing the normal cycle (8.9 ± 0.5 mm; p < 0.01) and also the value of P₄ in goats showing the short cycle (4.2 ± 2.1 ng/ml) was significantly lower than for those showing the normal cycle (10.3 ± 4.3 ng/ml; p < 0.05) at day 3 following ovulation. Three out of 5 Nubian goats became pregnant but only one goat carried to full term. The present experiment indicated that a combination of progesterone and eCG was effective in inducing estrus, although it resulted in a high incidence of short luteal lifespan. The low kidding rate and high incidence of embryonic loss may be due to the instability of the luteal lifespan.