Proteolytic activities in Phaseolus vulgaris cotyledons under copper stress

The changes in the protease activities of bean cotyledons were investigated in response to copper stress. Assays using synthetic substrates and specific protease inhibitors followed by activity measurements and electrophoresis analysis allowed to study the classes of enzymes involved in the storage protein mobilization during the germination of bean (Phaseolus vulgaris L) seeds, and then identify which ones were affected in the presence of 200 μM CuCl₂ in the imbibition medium. Copper treatment affected embryo growth and total protease activity. The results of SDS-gelatin-PAGE show that Cu excess led to a decrease in protease activity of 45 to 66 kDa. Moreover, cysteine-, aspartic- and metallo-protease activities were markedly lowered under copper stress, while serine-protease one was enhanced as well as its activity dependent abundance in comparison with control. However, the relative distribution of major cysteine protease in H₂O-germinated seeds was significantly diminished after Cu exposure. Thus, copper excess can disturb the nitrogen freeing from reserve tissues at enzymatic level; differential responses of protease classes are discussed, notably, cysteine protease in the way of storage protein mobilization and serine protease in protective mechanism one.     

Copper reduction by yeast cell wall materials and its role on copper uptake in Debaryomyces hansenii

Copper binding reducing activities of cell wall materials (CWM) prepared from cells of the yeast Debaryomyces hamsenii were examined. When CWM was treated with copper sulfate (0.1 mM CuSO₄), the copper was partially reduced from Cu (II) to Cu (I) and bound to CWM (below 10 nmol per mg dry wt.). The bound copper was mostly in the fraction of mannan-protein. Both copper-binding ability and protein content decreased with protease treatments. Mannan-protein prepared from CWM bound more copper than mannan did. This suggests that Cu (II) bound to the protein portion in CWM and was reduced to Cu (I). The optimum pH of copper reduction by CWM was about 5.0. The amount of copper bound to CWM increased with reducing agents and decreased with oxidizing agents. On the other hand, the copper uptake by yeast whole cells and spheroplasts was also stimulated by reducing agents, but inhibited by oxidizing agents. Furthermore, copper uptake by spheroplasts was stimulated in the presence of CWM. The optimum pH of copper uptake coincided with that of copper reducing activity. These results suggest that yeast cell wall not only supplies copper binding but also reduces copper, and the reduced copper is transported into yeast cells. The yeast cells may have copper-reducing proteins in the cell wall.     

饲料中铜水平对凡纳滨对虾免疫相关基因表达和抗菌能力的影响

在饲料中添加0、30和50 mg Cu/kg饲料的蛋氨酸铜,投喂凡纳滨对虾(Litopenaeus vannamei)7、14和21d,检测对虾体组织铜蓄积、免疫相关基因(Toll受体mRNA和溶菌酶mRNA)表达水平和免疫抗菌能力的变化。结果表明:凡纳滨对虾肝胰腺铜含量随着饲料蛋氨酸铜添加量增加及投喂时间延长而显著增加(P0.05);对虾肌肉的铜含量显著低于肝胰腺的铜含量。饲料中铜水平对凡纳滨对虾肌肉、血淋巴及肝胰腺中溶菌酶活性无显著影响(P0.05)。对虾组织SOD活性因饲料中铜水平和投喂时间变化显著,添加30 mgCu/kg组对虾肌肉、血淋巴和肝胰腺中SOD活性在第21天时显著高于其他两组(P0.05)。饲料中铜水平对凡纳滨对虾鳃组织中溶菌酶mRNA表达水平无显著影响,但显著影响鳃组织Toll受体mRNA表达水平(P0.05)。第7天时凡纳滨对虾Toll受体mRNA表达水平随着饲料铜水平升高而显著升高(P0.05);第14和第21天时,Toll受体mRNA表达水平在摄食添加30 mg Cu/kg组最高。人工急性感染溶藻弧菌(Vibrioalginolyticus)实验表明,第7天时,摄食添加50 mg Cu/kg组凡纳滨对虾全致死时间和半致死时间长于未添加铜组和添加30 mgCu/kg组,但在第14天,摄食添加30 mg Cu/kg组的全致死时间和半致死时间最长。研究表明饲料铜添加水平不但影响组织中铜的蓄积,还影响凡纳滨对虾SOD活性和Toll受体mRNA表达水平,从而影响机体的抗弧菌能力。     

Evaluation of Bioaccumulation and Toxic Effects of Copper on Hepatocellular Structure in Mice

The present study was to evaluate the hepatotoxicity effects in mice exposed to copper (Cu) used as dietary supplements for 95 days. Cu-treated mice showed increased body weight, and no toxic symptoms were observed at the beginning, but the tendency gradually changed with progress of experiment. In the liver, beneficial metals [Cu, iron (Fe), zinc (Zn), manganese (Mn), and molybdenum (Mo)] were analyzed by flame atomic absorption spectrometry. The content of Cu maintained at the same level during the experiments, but not resulting in the imbalance of Fe, Zn, Mn, and Mo being distributed. The activities of alkaline phosphatase (AKP) and super oxidation dismutase (SOD) showed significantly improvement during the first 30 days in Cu-supplemented group (P?<?0.01) but declined rapidly from 30th to 60th days, and later, they stabilized and were not statistically significant compared with control (P?>?0.05). No statistically significant correlation of ceruloplasmin (CPL) activity was appreciated during the experiment. The histopathological and ultrastructural abnormalities changes were observed in the liver of mice including vacuolar degeneration, necrosis, karyorrhexis, and endolysis. Many hepatocytes showed increased collagenic fibers, appearance of triglyceride droplets, and swollen mitochondria due to oral route of copper, which may lead to lipid peroxidation and free radicals. In conclusion, our study showed that exposure to copper influenced behavioral pattern and body weight, affected several enzymatic activities, and led to the physiological and considerable structural changes in the liver of mice. The public should pay more attention to avoid being exposed to copper.     

Zinc,copper, and zinc- or copper-dependent enzymes in human hypertension

Imbalance of zinc and copper status has been hypothesized in human hypertension. A case-control study was carried out to elucidate the possible relationship between zinc and copper status and essential hypertension. Thirty-one subjects affected by mild stable hypertension, pharmacologically untreated, were investigated together with 31 normotensive controls individually matched for sex, age, and smoking habits. Zinc and copper in serum and urine were measured, and serum activities of alkaline phosphatase (AP), lactic dehydrogenase (LDH), copper-zinc superoxide dismutase (Cu?Zn SOD), lysyl oxidase (LOX), and monoamine oxidase (MAO) were evaluated. No significant difference in serum and urine zinc and copper content as far as in serum activity of zinc (AP and LDH) or copper (Cu?Zn SOD, LOX, and MAO)-dependent enzymes was found between hypertensives and normotensives. Positive relationships were found in normotensives between serum and urine levels of zinc (r=0.577;p=0.001) and copper (r=0.394;p=0.028), and between serum copper and Cu?Zn SOD (r=0.534;p=0.002). In normotensives, diastolic blood pressure and serum zinc were positively related (r=0.370;p=0.041). In hypertensives, inverse correlations were observed between diastolic blood pressure and AP (r=?0.498;p=0.004) and Cu?Zn SOD (r=?0.452;p=0.011), and between systolic blood pressure and LOX (r=?0.385;p=0.033). Diastolic blood pressure was related to LDH inversely in hypertensives (r=?0.357;p=0.049) and positively in normotensives (r=0.457;p=0.010). In normotensives, diastolic blood pressure was inversely related with MAO (r=?0.360;p=0.046). These findings support the hypothesis that an imbalance of zinc and copper status might be involved in human hypertension.     

Effects of Different Copper Sources and Levels on Plasma Superoxide Dismutase, Lipid Peroxidation, and Copper Status of Lambs

This study was performed to determine the effects of different copper (Cu) sources and levels on plasma superoxide dismutase (SOD), lipid peroxidation, and Cu status of lambs. Fifty Dorper × Mongolia wether lambs (approximately 3?month of age; average BW?=?23.8?±?0.6?kg) were divided into five equal groups each with ten animals according to their weight. Treatments consisted of (1) control (no supplemental Cu), (2) 10?mg Cu/kg DM from Cu-lysine, (3) 20?mg Cu/kg DM from Cu-lysine, (4) 10?mg Cu/kg DM from tribasic copper chloride (Cu(2)(OH)(3)Cl; TBCC), and (5) 20?mg Cu/kg DM from TBCC. The Cu concentration was 6.74?mg/kg DM in the basal diet. Plasma copper concentrations and ceruloplasmin activities were not affected on day?30 by Cu supplementation. Copper supplementation increased plasma and liver copper concentrations and ceruloplasmin activities on day?60. Muscle Cu concentrations were not affected by Cu supplementation. There were no differences in plasma, liver, and muscle Cu concentrations and ceruloplasmin activities between Cu-lysine and TBCC. Liver copper concentrations and plasma ceruloplasmin activities were increased in lambs supplemented with 20?mg Cu/kg DM than in those supplemented with 10?mg Cu/kg DM on day?60. However, copper levels had no effects on Cu concentrations in plasma and muscle. Malondialdehyde (MDA) concentrations were decreased in plasma and liver tissues, but not affected in muscle by Cu supplementation. Plasma SOD activities were increased by Cu supplementation. There were no differences in plasma, liver, and muscle MDA concentrations and plasma SOD activities between Cu sources and levels. These results indicated that Cu supplementation increased plasma SOD activity, lipid oxidative stability, and copper status of lambs, but did not influence lipid oxidative stability in sheep muscle. Cu-lysine and TBCC were of similar availability when offered to finishing sheep.     

Regulation of copper uptake and transport in intestinal cell monolayers by acute and chronic copper exposure

Adaptation to high and low copper intake in mammals depends on the cellular control of influx, efflux and storage mechanisms of cellular copper concentrations. In the present study, we used an intestinal cell line (Caco-2), grown in bicameral chambers to study the effect of equilibrium loading with copper. We analyzed (64)Cu uptake from the apical surface, intracellular metal (Cu, Zn, Fe) content, (64)Cu transport into the basal chamber, and total copper, zinc and iron in the basal chamber. We found that the (64)Cu uptake is saturable, shows a linear response phase up to 1.5 microM reaching a plateau at 4-6 microM extracellular Cu. Intracellular copper increased 21.6-fold, from 1.5 to 32.4 mM (at 0.2-20.2 microM extracellular copper respectively). The time course for (64)Cu uptake and transport was linear when the cells were incubated with different copper concentrations. Uptake increased 10-fold when intracellular copper concentration was raised. Fluxes were lowest at 1.5 mM and highest at 32.4 mM Cu intracellular copper (2.03 and 20. 98 pmole (64)Cu insert(-1) h(-1), respectively). The apical-to-basolateral copper transfer rate was lower at 32.4 mM as compared to 1.5 mM intracellular copper (0.55-1.95 pmole (64)Cu insert(-1) h(-1), respectively). The total copper in the basal chamber increased 4.2-fold (from 3.04 to 12.85 pmole Cu insert(-1) h(-1)) when the intracellular copper concentration was raised. If cells are preincubated in a low copper medium most of the newly incorporated copper (64%) is transferred to the basolateral compartment. In contrast, under preloading with high copper concentration, only 4% of the fresh copper is transferred to the basal chamber; however, the intracellular copper contribution to this chamber increases by 4.2-fold. Thus, the process results in an increase in both storage and intracellular-to-basolateral flux of copper. In summary, our results indicate that copper fluxes from apical-to-cell and apical-to-basolateral domains are affected by intracellular copper concentration suggesting that mechanisms of copper transport involved in cellular adaptation to low and high copper exposure are different.     

The effects of copper, iron and zinc on digestive enzyme activity in the hybrid tilapia Oreochromis niloticus (L.) ×Oreochromis aureus (Steindachner)

The present experiment was conducted to study effects of Cu, Fe and Zn on activities of digestive enzymes of the hybrid tilapia Oreochromis niloticus×Oreochromis aureus. The acidic protease activities increased 65·5 and 55·1% by addition of homogenates of digesta‐containing stomach with copper (75 mg l⁻¹) and zinc (50 mg l⁻¹) respectively. Addition of Cu and Zn increased the activities of protease in the hepatopancreas homogenates by 132·7 and 38·1% respectively, and reduced the activity of protease in the digesta‐containing intestine homogenates by 11·0 and 13·8% respectively. Addition of Fe (50 mg l⁻¹) increased the acidic protease activity by 96·7% but did not alter the activities of protease in the intestine and hepatopancreas. Addition of Cu markedly inhibited activities of amylase in intestine and hepatopancreas homogenates, while Zn addition showed no effects. Addition of Fe reduced activities of amylase in the intestine homogenates by 47·9% but had no effect on amylase activities in the hepatopancreas. When Cu (75 mg kg⁻¹), Fe (50 mg kg⁻¹) and Zn (50 mg kg⁻¹) were supplemented to basal diet for 3 weeks, the activities of amylase in hepatopancreas homogenates increased 125·3, 215·6 and 70·0%, respectively, the activities of amylase in intestine increased 79·8, 74·6 and 48·5%, respectively, and the activities of lipase in intestine increased 90·5, 149·8 and 84·0%, respectively. Supplementation of Cu, Fe or Zn into diet had no effects on activity of protease in all digestive organs. Therefore, the results suggest that effects of Cu, Fe and Zn on activity of digestive enzymes in vitro were different from those seen in vivo, and that the positive effects of Cu, Fe and Zn supplemented to fish diet would be valuable information for formulating fish feed.     

Synthesis and structural analysis of the copper(II) complexes of N2-(2-pyridylmethyl)-2-pyridinecarboxamide

Previous investigations of the potential of metal-organic compounds as inhibitors of human immunodeficiency virus type I protease (HIV-1 PR) showed that the copper(II) complex diaqua [bis(2-pyridylcarbonyl)amido] copper(II) nitrate dihydrate and the complex bis[N2-(2,3,6-trimethoxybenzyl)-4-2-pyridinecarboxamide] copper(II) behaved as inhibitors of HIV-1 PR. In a search for similar readily accessible ligands, we synthesised and studied the structural properties of N2-(2-pyridylmethyl)-2-pyridinecarboxamide (L) copper(II) complexes. Three different crystal structures were obtained. Two were found to contain ligand L simultaneously in a tridentate and bidentate conformation [Cu(L(tri)L(bi))]. The other contained two symmetry-related ligands, coordinated through the pyridine nitrogen and the amide oxygen atoms [Cu(L(bi))(2)]. A search of the Cambridge Structural Database indicated that L(tri) resulting from nitrogen bound amide hydrogen metal substitution is favoured over chelation through the amide oxygen atom. In our case, we calculated that the conformation of L(tri) is 11 kcal/mol more favourable than that of L(bi). ESI-MS experiments showed that the Cu(L(bi))(2) structure could not be observed in solution, while Cu(L(tri)L(bi))-related complexes were indeed present. The lack of protease inhibition of the pyridine carboxamide copper(II) complexes was explained by the fact that the Cu(L(bi)L(tri)) complex could not fit into the HIV-1 active site.     

Antioxidants activities and concentration of selenium,zinc and copper in preterm and IUGR human placentas

The aim of this study was to examine changes in activities of cytochrome c oxidase (CCO), glucose-6-phosphate dehydrogenase (G6PDH), Cu–Zn superoxide dismutase (Cu–Zn SOD), glutathione peroxidase (GSH-Px), glutathione (GSH) levels and copper (Cu), zinc (Zn) and selenium (Se) concentrations, and to assess the possible differences between preterm placentas, placentas from term pregnancies complicated by intrauterine growth restriction (IUGR) and full-term control placentas.The enzyme activities and the level of GSH decreased in IUGR and preterm placentas in comparison with the control group. CCO activity and GSH level in preterm placentas were markedly lower compared with the IUGR (P<0.01; P<0.05) and control (P<0.01; P<0.05) placentas, respectively. In IUGR placentas the level of Cu was reduced by 23% (P<0.05) and Zn by 37%. In preterm placentas the level of Cu was reduced by 19% and Zn by 42%. Se level in IUGR and preterm placentas was higher (P<0.05) by 28% and 32% than in control group, respectively. The strong relation was observed between birth weight and CCO activity, birth weight and Cu–Zn SOD activity, and a low level of Zn and Cu influenced the birth weight especially in IUGR cases. Moreover, the strong inverse correlation between Se level and birth weight, Se level and placental weight and Se level and CCO activity are new findings.     

Source of Copper May Have Regressive Effects on Serum Cholesterol and Urea Nitrogen Among Male Fattening Lambs

An experiment was conducted to determine the effect of dietary copper (Cu) on mineral profile, hematological parameters, and lipid metabolism in lambs. Eighteen Zandi male lambs (approximately 3 months of age; 17.53?±?1.6 kg of body weight) were housed in individual pens and were assigned randomly to one of three treatments. Treatments consisted of (1) control (no supplemental Cu), (2) 10 mg Cu/kg dry matter (DM) from copper sulfate (CuS), and (3) 10 mg Cu/kg DM from Cu proteinate (CuP). The Cu concentration was 8.2 mg/kg DM in the basal diet. Blood was sampled from the jugular vein at the beginning of the study (enrollment, before feeding Cu supplement) and at days 25, 50, and 70 of experiment. The amounts of total serum glucose, urea nitrogen, calcium, phosphorus, iron, copper, zinc, and lipids and hematological parameters were measured. Average daily gain and feed efficiency were improved (P?<?0.05) with Cu supplementation and were better for the lambs fed diet supplemented with CuP. The concentrations of serum Ca, P, and Zn were not affected by source of Cu in the diet. However, Fe concentration was lower (P?<?0.01) in the Cu-supplemented groups. Experimental treatment had no significant effects on the hematological parameters. The serum glucose concentration was not affected by treatments. However, the urea nitrogen concentrations were significantly affected (P?<?0.05) by added Cu and was lower for CuP group as compared to the lambs in the CuS and control groups. Addition of Cu had no influence (P?>?0.05) on the serum triglyceride concentration, but lambs fed with CuP supplement had lower (P?<?0.05) serum cholesterol than the CuS and control animals. These results indicated that CuP supplemented at 10 mg/kg DM improved gain and enhanced the efficiency of nitrogen in male lambs.     

Heavy metal contamination in surface soils of the industrial district of Wuhan,China

The Qingshan district of Wuhan City is a typical Chinese industrial area. An increase in heavy metal pollution in the region's soil, due to industrialization and urbanization, has become a serious environmental problem. Surface soil samples from 155 sites were collected and analyzed. The median concentrations of cadmium (Cd), chromium (Cr), copper (Cu), lead (Pb), and zinc (Zn) in soil were 2.3, 46.2, 24.3, 28.2, and 86.8 mg/kg, respectively. Principal component analysis coupled with hierarchical cluster analysis showed that (1) in residential and industrial areas, Pb, Cu, and Zn concentrations were mainly affected by industrial emissions and traffic emissions, whereas in agricultural areas Cu and Zn were less affected by industrial emission and traffic emission, whereas Pb was affected by agricultural activities; and (2) Cd originated from a combination of sources, including industrial activities, traffic emission, and hypergene geochemical characteristics. The integrated pollution index varied from 1.1 to 16.6 with a mean of 3.9, and 70.6% of the area is extremely contaminated, 28.1% is heavily contaminated, and the remainder is moderately contaminated.     

茶(Camellia sinensis L.)对铜的吸收与累积

对广东省5个大型茶场中不同土壤背景值的8个茶园进行了土壤及茶(CamelliasinensisL.)树各部位的铜含量分析调查。地处我国南方的广东省茶园,土壤中铜含量在2.39～53.05mg kg之间。有效态铜含量相对较低,仅在0.45～3.17mg kg之间。茶树中各部位铜含量大小依次为枝条>幼叶>根>成熟叶。茶树枝条和幼叶铜含量较高,平均分别达14.5和14.3mg kg,远高于成熟叶铜含量水平(9.7mg kg)。而同时期相应茶场的茶叶制成品铜含量则较幼叶(一芽两叶)铜的含量高。土壤中的总钾、有效磷和总铜等都对茶树各部位铜的累积产生较大影响,但只有土壤有效磷含量对铜在茶树体内向枝条部分的转移和累积影响达显著程度,枝条铜含量与土壤有效磷之间的相关系数r=-0.74,呈显著负相关(P<0.05)。     

Influence of Dietary Copper Methionine Concentrations on Growth Performance,Digestibility of Nutrients,Serum Lipid Profiles,and Immune Defenses in Broilers

A 42-day experiment was conducted to evaluate the influence of dietary copper (Cu) concentrations on growth performance, nutrient digestibility, and serum parameters in broilers aged from 1 to 42 days. Five hundred forty 1-day-old broilers were randomly assigned into 1 of the following 6 dietary treatments: (1) control (basal diet without supplemental Cu), (2) 15 mg/kg supplemental Cu (Cu15), (3) 30 mg/kg supplemental Cu (Cu30), (4) 60 mg/kg supplemental Cu (Cu60), (5) 120 mg/kg supplemental Cu (Cu120), and (6) 240 mg/kg supplemental Cu (Cu240), Cu as copper methionine. A 4-day metabolism trial was conducted during the last week of the experiment feeding. The results showed that dietary Cu supplementation increased the average daily gain and the average daily feed intake (P < 0.01). The feed gain ratio, however, was not affected by dietary Cu (P > 0.10). Additionally, dietary Cu supplementation increased the digestibility of fat and energy (P < 0.05). The concentration of serum cholesterol, triglycerides, and high-density lipoprotein cholesterol decreased with dietary Cu supplementation (P < 0.05). The activities of serum Cu-Zn superoxide dismutase (P < 0.05), glutathione peroxidase (P < 0.05), and ceruloplasmin (P = 0.09), on the contrary, were increased by Cu addition. For immune indexes, dietary Cu supplementation increased serum IgA and IgM (P < 0.05). In addition, the activities of serum ALT increased with increasing dietary Cu supplementation (P < 0.05). In conclusion, our data suggest that Cu supplementation can increase fat digestibility and promote growth. Additionally, dietary Cu supplementation can reduce serum cholesterol and enhance antioxidant capacity in broilers.

     

Physiological responses of <Emphasis Type="Italic">Lupinus luteus</Emphasis> to different copper concentrations

Yellow lupin (Lupinus luteus L.) plants were grown in hydroponic solution for 15 d under different copper concentrations (0.1, 0.5, 1.0, 10, 25 and 50 μM). With increasing Cu concentration total biomass was not affected, leaf area slightly decreased, while chlorophyll content decreased considerably. Cu content increased significantly both in roots and in leaves, but the contents of other ions were only slightly affected at the highest Cu concentration (Mn content decreased both in roots and in leaves, P content decreased only in leaves and Zn content increased in roots). Superoxide dismutase (SOD) activity increased up to day 7 after copper application. Peroxidase (GPOD) and polyphenol oxidase (PPO) activities also increased, while catalase (CAT) activity remained constant.     

Crystal structures of multicopper oxidase CueO bound to copper(I) and silver(I): functional role of a methionine-rich sequence

The multicopper oxidase CueO oxidizes toxic Cu(I) and is required for copper homeostasis in Escherichia coli. Like many proteins involved in copper homeostasis, CueO has a methionine-rich segment that is thought to be critical for copper handling. How such segments function is poorly understood. Here, we report the crystal structure of CueO at 1.1 Å with the 45-residue methionine-rich segment fully resolved, revealing an N-terminal helical segment with methionine residues juxtaposed for Cu(I) ligation and a C-terminal highly mobile segment rich in methionine and histidine residues. We also report structures of CueO with a C500S mutation, which leads to loss of the T1 copper, and CueO with six methionines changed to serine. Soaking C500S CueO crystals with Cu(I), or wild-type CueO crystals with Ag(I), leads to occupancy of three sites, the previously identified substrate-binding site and two new sites along the methionine-rich helix, involving methionines 358, 362, 368, and 376. Mutation of these residues leads to a ∼4-fold reduction in k_cat for Cu(I) oxidation. Ag(I), which often appears with copper in nature, strongly inhibits CueO oxidase activities in vitro and compromises copper tolerance in vivo, particularly in the absence of the complementary copper efflux cus system. Together, these studies demonstrate a role for the methionine-rich insert of CueO in the binding and oxidation of Cu(I) and highlight the interplay among cue and cus systems in copper and silver homeostasis.     

The influence of heavy metals upon the growth of sitka-spruce in South Wales forests

Summary Experiments on sitka-spruce seedlings grown in acidic peaty gley soils under green-house conditions, where the soils where doped with increasing amounts of Cd, Cu and Pb up to maximum levels of metal added of 16 ppm, 32 ppm and 400 ppm respectively, showed that the levels of Cd and Pb in shoots and roots increased with increasing levels in the soil, whereas levels of copper appeared to be independent. The addition of these three metals to the soils did not influence the uptake of other heavy metals, or of the nutrients potassium or calcium. Increases in the shoot cadmium levels significantly reduced the yields of the plant shoots. However, the plant yields were only affected by the highest level of lead that was added to the soil (400 ppm Pb) and unaffected by all the copper treatments (0–32 ppm Cu in the soil). The lengths of the sitka-spruce roots were reduced when cadmium and lead levels in the soil exceeded certain threshold concentrations (2.5 ppm total Cd, where 0.3 ppm was extractable with 0.5 M acetic acid; and 48 ppm total Pb, where 1.7 ppm was extractable). However, root lengths were not reduced by copper. This was probably related to the fact that copper appears to be relatively unavailable in the type of soil used, as only 1.1. ppm Cu was extractable from a total of 32 ppm Cu added. Root branching was apparently reduced by increases in the soil levels of cadmium, copper and lead. The roots of some control plants had symbiotic mycorrhizal associations (4 out of 19 plants), whereas the roots of all the plants grown in the soils with added heavy metals did not develop these.     

Copper and zinc fractions affecting microorganisms in long-term sludge-amended soils

The influences of Zn and Cu on soil enzyme activities (acid phosphatase, alkaline phosphatase, arylsulfatase, cellulase, dehydrogenase, protease (z-FLase), urease, beta-D-glucosidase and beta-D-fructofuranosidase (invertase)) and microbial biomass carbon were investigated in agricultural soils amended with municipal sewage sludge or compost since 1978. The trace metals in the soils were fractionated using a sequential extraction method. Long-term application of the sewage sludge and composts caused accumulations of Cu and Zn in the soils, ranging from 140 to 144 and from 216 to 292 mg kg(-1), respectively. The percentage of Cu was highest in the NaOH- and HNO3-extractable fractions (44-51% and 38-46%, respectively), while the percentage of Zn was highest in the HNO3- and EDTA-extractable fractions (65-83% and 11-32%, respectively). Although the percentage of the bioavailable fractions (sum of KNO3 + H2O-, NaOH-, and EDTA-extractable amounts) of Cu (53-64%) was higher than that of Zn (15-37%), the percentage of the most labile fractions (KNO3 + H2O) of Zn (2.1-5.9%) was larger than that of Cu (1.1-2.4%). The size of the microbial biomass carbon increased with the application of sewage sludge or compost. For some enzymes, however, the ratio of the enzyme activity to microbial biomass was lower in the soils amended with sewage sludge or compost than that in the control soil. The soil enzyme activities were more adversely affected by Zn than by Cu. From a multiple regression analysis, it was found that dehydrogenase, urease, and beta-D-glucosidase activities were reduced by the KNO3 + H2O-extractable fraction of Zn in the soils. These microbial activities seem to be sensitive to Zn stress, indicating the possibility that they might be useful bioindicators for evaluation of the toxic effects of Zn on microorganisms in the soils.     

AtCCS is a functional homolog of the yeast copper chaperone Ccs1/Lys7

In plant chloroplasts two superoxide dismutase (SOD) activities occur, FeSOD and Cu/ZnSOD, with reciprocal regulation in response to copper availability. This system presents a unique model to study the regulation of metal-cofactor delivery to an organelle. The Arabidopsis thaliana gene AtCCS encodes a functional homolog to yeast Ccs1p/Lys7p, a copper chaperone for SOD. The AtCCS protein was localized to chloroplasts where it may supply copper to the stromal Cu/ZnSOD. AtCCS mRNA expression levels are upregulated in response to Cu-feeding and senescence. We propose that AtCCS expression is regulated to allow the most optimal use of Cu for photosynthesis.     

Effects of Different Sources and Levels of Copper on Growth Performance,Nutrient Digestibility,and Elemental Balance in Young Female Mink (Mustela vison)

An experiment was conducted in a 3?×?3?+?1 factorial experiment based on a completely randomized design to evaluate the effects of different sources of copper on growth performance, nutrient digestibility and elemental balance in young female mink on a corn–fishmeal-based diet. Animals in the control group were fed a basal diet (containing 8.05 mg Cu/kg DM; control), which mainly consisted of corn, fish meal, meat bone meal, and soybean oil, with no copper supplementation. Minks in other nine treatments were fed basal diets supplemented with Cu from reagent-grade copper sulfate, tribasic copper chloride (TBCC) and copper methionate. Cu concentrations of experiment diets were 10, 25, and 40 mg/kg copper. A metabolism trial of 4 days was conducted during the last week of experimental feeding. Final body weight and average daily gain increased (linear and quadratic, P?<?0.05) as Cu increased in the diet; maximal growth was seen in the Cu25 group. Cu supplementation slightly improved the feed conversion rate (P?=?0.095). Apparent fat digestibility was increased by copper level (P?=?0.020). Retention nitrogen was increased by copper level (linear, P?=?0.003). Copper source had a significant effect on copper retention with Cu-Met and copper sulfate treatments retention more than TBCC treatments (P?<?0.05). Our results indicate that mink can efficiently utilize added dietary fat and that Cu plays an important role in the digestion of dietary fat in mink, and mink can efficiently utilize Cu-Met and CuSO₄.