Intracellular distribution of copper and zinc in the liver of copper-exposed cattle from northwest Spain

The intracellular distribution of copper (Cu) and zinc (Zn) in the livers of normal and moderately Cu-exposed cattle was investigated with the aim of improving understanding of the pathophysiology of Cu accumulation in cattle. At total liver Cu concentrations within the generally accepted normal range (25-100 mg/kg fresh weight) the large-granule fraction was the main subcellular compartment for Cu accumulation, followed by the cytosol and the nucleus, whereas in the microsomal fraction Cu accumulation was very low. With increasing Cu exposure, the capacity of the large-granule fraction to accumulate Cu decreased, proceeding towards a plateau (estimated at about 80 mg/kg at a projected total liver Cu concentrations of 450 mg/kg), accompanied by progressively greater Cu accumulation in the nucleus and cytosol. Total liver Cu concentration had little influence on subcellular Zn distribution, with hepatic Cu concentrations being only moderately correlated with Zn concentration in the large-granule fraction. There was a strong association between the proportion of total Cu bound to metallothionein (MT) and the proportion of total Zn in the large-granule fraction, suggesting that Zn displaced from MT is taken up by the lysosomes for excretion. This pattern of Cu accumulation, as in sheep, may be due to the limited capacity for metallothionein binding of Cu and excretion in bile.     

Assessment of some blood parameters as potential markers of hepatic copper accumulation in cattle.

The increasing frequency of copper (Cu) toxicosis episodes in cattle in recent years, mainly associated with excess Cu supplementation, underscores the need to identify animals in the clinically silent phase of hepatic Cu accumulation. The aim of the study reported here was to evaluate the suitability of various blood parameters as potential early markers of hepatic Cu accumulation in cattle. Paired liver and blood samples from 70 calves aged 6 to 10 months were obtained at slaughter in a region in northwestern Spain where animals usually have hepatic Cu concentration higher than safe values. Neither serum Cu concentration nor ceruloplasmin (CP) concentration, the 2 parameters most commonly used for diagnosis of Cu deficiency, were significantly associated with hepatic Cu concentration. However, whole-blood Cu concentration had a slight but significant correlation with hepatic Cu concentration (r = 0.269, P = 0.026). The use of calculated blood parameters, such as the serum or whole-blood non-CP Cu fraction, or the CP-to-serum Cu ratio, increased the correlation with the hepatic Cu concentration (r = 0.393, P = 0.001, in the case of whole-blood, non-CP Cu concentration), but the strength of the association remained insufficient for accurate prediction of hepatic Cu values. Likewise, hepatic enzyme (aspartate transaminase and gamma-glutamyltransferase) activities were significantly or nearly correlated significantly with hepatic Cu concentration, but the strength of the association remained too low for accurate prediction. It is concluded that direct analysis of Cu concentration in liver biopsy specimens--preferably postmortem liver samples obtained at slaughter--is the best technique currently available for detecting chronic subclinical Cu accumulation in cattle at risk for supraoptimal Cu exposure.     

Breed differences in sheep with respect to the interaction between zinc and the accumulation of copper in the liver

In sheep the effects of Zn‐supplementation on the Cu‐accumulation in the liver were studied.

At a relatively high Cu‐level in the feed, dietary Zn reduced the hepatic Cu‐level in the Texel breed and in the crossbreed Texel x Friesian milksheep, but not in the Friesian milksheep breed.

On account of this result it can be concluded that in sheep the mutual interaction between Cu and Zn may be dependent on genetic factors.     

Mineral deficiency status of ranging zebu (Bos indicus) cattle around the Gilgel Gibe catchment, Ethiopia

Mineral deficiencies in cattle, widespread in East Africa, impair optimal health and production and consequently place a great burden on the farmers' income. Therefore, detection of shortages and imbalances of specific minerals is essential. Our objective was to evaluate the mineral status of grazing cattle around the Gilgel Gibe catchment in Ethiopia and associated factors. In study I, individual animal plasma and herd faecal Ca, P, Mg, Na, K, S, Fe, Zn, Mn and Cu concentrations were determined in adult zebu cattle (Bos indicus; n?=?90) grazing at three altitudes around the catchment, whilst recording body condition score and sex. In study II, liver samples of adult male zebu cattle (n?=?53) were analysed for Cu, Zn, Fe, Se and Mo concentrations and inspected for parasitic infections. Plasma and liver analyses revealed a Cu deficiency problem in the area, since 68 and 47 % of cattle, respectively, were Cu deprived according to diagnostic criteria for Bos taurus cattle. High hepatic Mo concentrations in 17 % of cases might reflect excessive dietary Mo intake. Liver Se and plasma Na concentrations were too low in 92 and 80 % of cattle. Plasma Mn concentrations were largely below the detection limit. Plasma Cu as well as Ca concentrations were lower in the lowest altitude compared to the highest altitude group (P?<?0.05), whereas lean to medium cattle had lower plasma Cu concentrations (P?<?0.05). No differences in hepatic mineral concentrations were detected between cattle with different types of parasitic infection. In conclusion, bovine mineral deficiencies were present in the Gilgel Gibe area and were associated with grazing altitude and body condition score.     

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.     

Heavy metals in fluids and tissues of fetal calves and in young calves of nursing cows exposed or not exposed to anaerobically digested wastewater sludge

Fetal fluid, diaphragm muscle, heart, kidney, and liver from 14 four- to five-month-old calf fetuses were analyzed for Zn, Cu, Cd, Cr, Ni, Pb, and Fe. The same tissues and bone were examined from postparturient calves 30, 60, and 120 days of age. Weaned calves (8 to 9 months old) were necropsied and diaphragm muscle, heart, kidney, liver, brain, and bone were analyzed. Additional calves from cows exposed or not exposed to anaerobically digested sludge were placed in a feed yard at weaning for a feeding period of 150 days and then were killed and necropsied. Heavy metal content of the aforementioned tissues was compared with the same tissues obtained from peer calves weaned, killed, and necropsied 150 days earlier. Zinc, Cu, and Fe were up to 11 times higher in fetal livers than in control or exposed postparturient calves. These concentrations declined after birth and stabilized at approximately the concentrations observed in adult cattle at about 12 to 14 months of age. Cadmium accumulated to a greater extent in kidneys and livers of exposed calves than in control calves. However, the accumulation did not produce a detectable effect upon the functional or morphologic characteristics of the organs examined. Other metals were present at low levels and were generally not remarkably different in control and exposed animals.     

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.     

Yellow-necked mice (Apodemus flavicollis) and bank voles (Myodes glareolus) as zoomonitors of environmental contamination at a polluted area in Slovakia

Background

Free-living wild rodents are often used as zoomonitors of environmental contamination. In the present study, accumulation of cadmium (Cd), copper (Cu), iron (Fe), and zinc (Zn) in critical organs of yellow-necked mice (Apodemus flavicollis) and bank voles (Myodes glareolus) trapped in a polluted area in Nováky, Slovakia was investigated.

Methods

Yellow-necked mice (n = 8) and bank voles (n = 10) were collected using standard theriological methods for wood ecosystems. All animals were adult males in good physical condition. The concentrations of Cd, Cu, Fe, and Zn in the liver, kidney, and bone were determined by atomic absorption spectrophotometry.

Results

The highest concentrations of Cd and Zn were found in the bone of both species while Cu and Fe accumulated mainly in kidney or liver. Significant higher concentrations of Cd and Cu were detected in the liver of bank voles than in yellow-necked mice. Similar significant higher levels of Cd and Zn were found in the bone of bank voles. In contrast, significant higher concentrations of Cu and Fe were present in the kidney of yellow-necked mice.

Conclusions

In the yellow-necked mouse and bank vole, bone seems to accumulate Cd and Zn following prolonged exposure. On the contrary, kidney and liver store Cu and Fe after a long-term environmental exposure. In the present study, bank voles seemed to be more heavy metal loaded zoomonitors than yellow-necked mice.     

Mineral concentrations of plasma and liver after injection with a trace mineral complex differ among Angus and Simmental cattle

To examine the effects of cattle breed on the clearance rate of an injectable mineral product, 10 Angus and 10 Simmental steers were blocked by breed and initial BW (332 ± 33 kg) and injected with either Multimin 90 (MM) or sterilized saline (CON) at a dose of 1 mL/45 kg BW. Multimin 90 contains 15 mg Cu/mL (as Cu disodium EDTA), 60 mg Zn/mL (as Zn disodium EDTA), 10 mg Mn/mL (as Mn disodium EDTA), and 5 mg Se/mL (as sodium selenite). Steers received a corn-silage-based diet, and inorganic sources of Cu, Zn, Mn, and Se were supplemented at NRC recommended amounts. Jugular blood was collected immediately before injection and at 8 and 10 h post-injection and on days 1, 8, and 15 post-injection. Liver biopsies were collected 3 d before injection and on days 1, 8, and 15 post-injection. Liver and plasma mineral concentration and glutathione peroxidase (GSH-Px) activity data were analyzed as repeated measures. Plasma concentrations of Zn, Mn, and Se were greater (P = 0.01) and Cu tended to be greater (P = 0.12) post-injection in MM steers compared with the CON steers. Regardless of treatment, Simmental cattle had lower plasma concentrations of Cu, Zn, and Se (P ≤ 0.05) when compared with Angus cattle. Erythrocyte GSH-Px activity was greater (P = 0.01) in MM steers compared with CON steers. Liver concentrations of Cu, Zn, and Se were greater (P = 0.05) in MM steers compared with CON steers post-injection. Liver Mn concentrations tended to be greater (P = 0.06) in MM steers compared with CON steers in the days post-injection. Interestingly, Simmental cattle exhibited greater (P = 0.01) liver Mn concentrations in the days after injection compared with Angus cattle (7.0 and 6.0 mg Mn/kg for Simmental and Angus cattle, respectively), regardless of treatment. It is unclear if this breed difference is biologically relevant; however, these data may suggest that differences in liver excretion of Mn exist between the two breeds. Overall, use of an injectable trace mineral increased liver concentrations of Cu and Se through the 15-d sampling period, suggesting that this injectable mineral is an adequate way to improve Cu and Se status of cattle through at least 15 d.     

The Status of Selected Minerals in Soil,Forage and Beef Cattle Tissues in a Semi-arid Region of Zimbabwe

Five districts in the Matabeleland region, an arid western area of Zimbabwe, were investigated for the status of Ca, P, Na, Cu and Zn in soil, forage and cattle during the wet and dry seasons over a period of one year. The cattle came from the natural grazing lands and were not supplemented at the time of sampling. Some deficiencies in soil Zn and P were found in the districts of Lupane and Bulilima-mangwe, respectively. Dry season soil Ca, Cu and P concentrations were significantly higher (p < 0.05) than rainy season values owing to leaching in all five districts. Most forage samples had mineral concentrations below the critical levels known to be adequate for animal requirements. Forage levels of Ca, Na, Cu and Zn significantly increased (p < 0.05) with advancing maturity, while P significantly decreased (p < 0.05) in almost all the districts. Marked deficiencies of minerals were found in cattle tissues and these levels followed the seasonal trend seen in the forage. These results indicate that cattle in Matebeleland are deficient in P, Ca, Cu and Zn and that grazing areas in the region cannot provide adequate levels of the five minerals studied.     

The effect of pig farming on copper and zinc accumulation in cattle in Galicia (north-western Spain)

Copper and zinc are frequently added at high concentrations to pig diets as growth promoters. Livestock grazing pasture contaminated with pig slurry may, therefore, be at risk from excessive intake of these elements. High liver copper concentrations have been detected in cattle from the agricultural region of Galicia (NW Spain), especially where there is intensive pig farming. The aim of this study was to evaluate whether pig farming does affect accumulation of copper and zinc in cattle in Galicia. Hepatic copper and zinc concentrations in calves were elevated in areas with naturally high levels of these elements in the soil. The densities of young pigs (piglets and growing-finishing pigs), but not reproductive sows, also influenced copper accumulation in calves. Liver copper levels in calves were significantly and positively related to the density of young pigs in the region. In areas with the highest pig densities, more than 20% of the cattle analysed had hepatic copper concentrations that exceeded the potentially toxic concentration of 150 mg/kg fresh weight. There was no evidence that zinc accumulation in calves was affected by pig density.     

Clinical observations of cattle and buffalos with experimentally induced chronic copper poisoning

The susceptibility of cattle and buffalos to chronic copper poisoning (CCP) was compared by using cattle (n = 10) and buffalo (n = 10) steers distributed into two copper supplemented (n = 6) and two control (n = 4) groups. Supplemented animals received 2 mg copper (Cu)/kg body weight daily for one week, with an additional 2 mg weekly until the end of the experiment (day 105). Three liver biopsies (day 0, 45, and 105) were obtained for mineral analyses; clinical examinations and blood samples were obtained every 15 days. Three supplemented cattle and two buffalos with typical manifestations of CCP died. There were no differences in the frequency of mortality between cattle and buffalos; hepatic copper concentration was higher in cattle than buffalos. These findings suggest that buffalos and cattle might be equally susceptible to CCP. However, buffalos accumulate less liver copper than cattle and have a lower threshold of hepatic Cu accumulation, which leads to clinical manifestation of CCP.     

Clinical zinc and copper deficiencies in cattle of western Sudan

Clinical cases of both Zn and Cu deficiencies are reported in a cattle farm in Kordofan Region of the Sudan after drought. The animals showed general weakness, stunted growth, infertility, parakeratosis and achromotrichia. There was macrocytic hypochromic anaemia and low Cu and Zn concentrations in sera. The condition was more prevalent in zebu-Friesian crosses than the local breeds. Drought and marginal or low Cu and Zn content in pasture may be the predisposing factors.     

Ultrastructural and kinetic studies of copper metabolism in Bedlington Terrier dogs

The objectives of this study were to determine the time course of copper (Cu) absorption in the Bedlington Terrier (BT) dog after duodenal infusion of Cu and to determine the onset of Cu accumulation in BT puppies. At 80 minutes after infusion, Cu was present in the bile and liver at 26.1 and 2675 microg/g dry weight (dw), as compared with 598 and 224 microg/g dw, respectively, in the control. In the puppies, the mean liver Cu concentrations were 443 +/- 216 microng/g dw as compared with 54 +/- 34 microg/g dw in their controls. The BT puppies had large numbers of Cu granules in their hepatic lysosomes, with very few in the bile canaliculi. In contrast, the controls had few granules in their lysosomes and large numbers in their canaliculi. These results suggest that hepatic Cu accumulation occurs rapidly and in utero in the BT.     

Influence of copper supplementation on blood and liver copper in cattle fed tall fescue or quackgrass

Copper concentrations were measured in naturally Cu-deficient and Cu-supplemented yearling Hereford heifers fed tall fescue (Festuca arundinacea Schreb.) or quackgrass (Agropyron repens [L] Beauv.) forage. The data were used to establish a relationship between dietary Cu supplementation and accumulation of liver Cu, and to test the influence of diet and Cu supplementation on blood plasma Cu and ceruloplasmin oxidase activity. Dietary Cu supplementation was positively correlated with accumulation of liver Cu; the rate of increase in liver Cu levels diminished at high supplementation rates (liver Cu = 410.43 - 353.47 x e-.00295 x mg Cu supplement; R2 = .740), suggesting an efficient mechanism preventing fast, toxic Cu-overloading in cattle. The linear relationship (R2 = .7878) between blood plasma Cu and ceruloplasmin oxidase activity indicated that similar percentages of blood plasma Cu were present as ceruloplasmin in deficient, normal and Cu-supplemented cattle. However, the portion of plasma Cu occurring as ceruloplasmin changes significantly with type of feed. When compared with quackgrass, tall fescue not only produced rapid Cu depletion in unsupplemented cattle, but affected intermediary Cy metabolism and activity of Cu-enzyme ceruloplasmin differently at all levels of experimental Cu supplementation.     

Interrelationships of exogenous porcine growth hormone administration and feed intake level affecting various tissue levels of iron, copper, zinc and bone calcium of growing pigs

Trace mineral status was evaluated in a 2 x 3 factorial treatment array with a total of 34 barrows growing from 25 to 55 kg live weight. Treatments included three levels of feed intake (100, 80 and 60% of ad libitum intake) and exogenous pituitary growth hormone (pGH) therapy (0 and 100 micrograms/kg BW daily). Blood was collected prior to slaughter for the determination of hematocrit and serum trace metal concentrations; tissues (liver, heart, kidney, bone and muscle) were obtained when pigs were slaughtered at 55 kg. Hematocrits and serum Fe were lower in pGH-treated pigs than in controls at all levels of feed intake. Serum Cu was increased by feed restriction but was not altered by pGH therapy. The concentration of serum Zn was not affected by either treatment. Concentrations of hepatic Fe and Cu were lower in pGH-treated pigs than in controls but were higher in feed-restricted pigs than in ad libitum-fed pigs. However, the total amounts of hepatic Fe and Cu were similar in pGH-treated pigs to concentrations in controls. The concentration of hepatic Zn was not influenced by either pGH treatment or feed intake. Femur weights were marginally greater in pGH-treated pigs, probably due to elevated water content. Iron concentration in bone was higher in pGH-treated pigs than in control pigs, whereas Ca, Cu and Zn were not influenced by pGH treatment or feed restriction. Feed intake and pGH treatment did not influence the concentrations of Fe, Cu or Zn in muscle. These findings indicate that pGH therapy affects the metabolism of Fe but has little impact on the overall composition of body ash.     

Trace mineral status and toxic metal accumulation in extensive and intensive pigs in NW Spain

The aim of the present study was to determine trace element status and toxic metal accumulation in extensive and intensive pigs in NW Spain. Soil, feed and animal tissues (liver, kidney and muscle from 112 pigs) were collected from extensive and intensive pig farms across NW Spain and analysed for metals by ICP-MS and ICP-OES. Our results indicate that animals from both extensive and intensive systems showed an adequate mineral status and that toxic metal residues were generally low, no sample exceeding the maximum admissible levels in meat and meat products established by the EU. Trace element concentrations were generally higher in the intensive pigs possibly reflecting mineral supplementation in the concentrate feed. The statistically significantly higher As, Hg and Pb residues as well as the better Fe and Ni status in extensive pigs could be related at least in part with soil ingestion when rooting. On the contrary, the higher Cd accumulation in the intensive pigs can be explained considering Cu and Zn supplementation in the concentrate feed. Our results also seem to indicate that toxic metal accumulation as well as trace element status in different tissues could be related to metabolic aspects (i.e. related to breed and growth rate) as well as certain management practices (i.e. physical exercise when grazing).     

Heavy metals in tissues of cattle exposed to sludge-treated pastures for eight years

Cattle allowed to graze pastures variously treated with anaerobically digested sludge (ADS) for up to 8 years, contained higher concentrations of heavy metals and organic compounds in some tissues than did cattle not exposed to ADS. Chemical analyses were made on heart, diaphragm, liver, kidney, brain, and bone to determine the amount of Zn, Cu, Cd, Cr, Ni, Pb, Fe, and Hg. Determinations for organic compounds (polychlorinated-biphenyls, dieldrin, and heptachlor/epoxide) were made on visceral fat. Cadmium was the only metal to accumulate consistently in increased amounts. Amounts of Cd accumulated in kidneys and livers were higher in cattle exposed to ADS than in cattle not exposed to ADS. Although the amounts were higher in cattle exposed to ADS, none of the cattle had detectable evidence of disease or pathologic conditions. The mean amount of Cd in kidneys of cattle exposed to ADS was 44 vs 9 mg/kg dry weight. Amounts of other heavy metals in the body tissues were generally not unusual and were similar in both groups of cattle. Polychlorinated-biphenyl concentrations were higher in the visceral fat of cattle exposed to ADS. All cattle in both herds remained healthy, and pathologic changes that could be attributed to association with ADS were not detected. Seemingly, ADS can be used safely to produce cattle forage without endangering the health of cattle.     

Variations in heavy metal concentrations among trophic levels of the food webs in two agroecosystems

Excessive accumulation of trace metal in soil represents a growing environmental problem posing severe risks to biota, humans and ecosystems. Concentrations of Cd, Pb, Cu and Zn were determined in soil, as well as in representatives of some trophic levels in the terrestrial food webs in two Egyptian agroecosystems; El-Manzala (a rural area located in the northeast of the River Nile Delta) and El-Tebbin (an industrial area located in South Cairo). Representatives of trophic levels included arthropods, amphibians, reptiles, birds and small mammals. Metal concentrations were determined in the leaves of wild plants, bodies of arthropods and livers of vertebrates. Levels of metals in the soil, plants and most animal species were higher in El-Tebbin than El-Manzala. Minimal concentrations of metals were detected more often in plants and in the cattle egret, whereas maximum values were common in the soil, amphibians and a mantid. Accumulation of metals was more frequent in arthropods and other taxa at lower trophic levels of food webs, suggesting that the transfer of metals along the vertebrate food web may be of relatively minor importance. However, of all the metals examined, only Pb was bioaccumulated to any appreciable extent in some of the higher trophic levels. A significant increase in liver mass and relative liver mass of the Norway rat from the polluted site was observed.     

泌阳县肉牛养殖常用饲料原料中微量元素及重金属含量分析

[目的]饲料原料中微量元素和重金属的含量对肉牛的生长发育及生态环境均有重要影响,通过对肉牛养殖中常用饲料原料中的微量元素及重金属含量进行分析,为肉牛养殖提供参考.[方法]研究对泌阳县境内24份肉牛养殖常用的饲料原料样品进行分析,采用硼氢化钾还原原子荧光光谱法测定了饲料原料中4种微量元素(铜、铁、锌、硒)及2种重金属(砷...