Copper Deficiency Induces Oxidative Stress in Liver of Mice by Blocking the Nrf2 Pathway.

Copper (Cu) is an essential metal required for many physiological processes and biological reactions. Liver is the main organ of metabolism of Cu and is also the site where synthesis of some metalloproteins. The purpose of this study is to explore the effects of Cu deficiency on the liver and to evaluate the changes in liver oxidative stress levels to reveal its possible impact mechanisms. Mice were feed to a nutritional Cu-deficiency diet from weaning and injected with copper sulfate (CuSO4) intraperitoneally to correct Cu deficiency. Cu deficiency resulted in reduced liver index, liver histological alteration, and oxidative stress; decreased the contents of Cu and ALB; elevated ALT and AST concentrations in serum together with decreased mRNA and protein expressions of Nrf2 pathway related molecules (Nrf2, HO-1, NQO1); and increased mRNA and protein expressions of Keap1. However, the supplement of copper sulfate (CuSO4) significantly ameliorated the changes mentioned above. Our results indicate that Cu deficiency can cause hepatic damage in mice is associated with the activation of oxidative stress and inhibition of Nrf2 pathway.

Effects of Various Copper Sources and Concentrations on Performance, Skeletal Growth, and Mineral Content of Excreta in Broiler Chickens.

The experiment was designed to study the effect of supplemental sources and concentrations of copper on the performance and development and mineralization of tibia bones in broiler chickens. A 42-day feeding experiment was conducted utilising three copper sources, including copper sulphate (CuS), copper chloride (CuCl), and copper propionate (CuP), each with four different concentrations, i.e. 8, 100, 150, and 200 mg/kg. The body weight gain with 200 mg Cu/kg food was noticeably higher during the first 4-6 weeks of age. Due to the interaction between Cu sources and levels, there was no significant change in the body weight gained. The feed intake during various growing phases did differ significantly neither the main effect nor the interaction between different copper sources and levels. A CuP-supplemented diet (200 mg/kg food) considerably (P ≤ 0.05) improved the feed conversion ratio between 4-6 and 0-6 weeks. At the end of the experiment, a total of 72 tibia bones, i.e. six for each treatment were collected. A metabolic trial was conducted to look into mineral retention in broiler chickens on the final 3 days of the trial (40-42 days). Increased tibia bone zinc (Zn) levels were seen with the addition of 8 mg Cu/kg of Cu chloride, 100 mg Cu/kg of Cu propionate, 8 mg Cu/kg of Cu sulphate, and 8 mg/kg of Cu propionate to the diet. At higher levels of Cu (150 and 200 mg/kg diet), there was a significantly (P ≤ 0.01) reduced tibia Zn content. Cu sulphate treatment group had higher (P ≤ 0.01) tibia Cu content (8 mg Cu/kg diet). Cu sulphate supplemented diet had a greater excreta Zn content (P ≤ 0.01) than Cu chloride supplemented diet, and Cu propionate supplemented diet had the lowest excreta Zn content. Excreta with a higher Fe concentration were found in diets supplemented with copper sulphate and copper chloride (P ≤ 0.05) than in diets supplied with copper propionate. Thus, it may be concluded that feeding dietary Cu concentrations up to 200 mg Cu/kg diet, regardless of the different sources, had no negative effects on bone morphometry and mineralization parameters with the exception of a decrease in the tibia's zinc content.

Vitamin D alleviates cognitive dysfunction and brain damage induced by copper sulfate intake in experimental rats: focus on its combination with donepezil.

This study aimed to demonstrate the potential benefits of donepezil (DPZ) and vitamin D (Vit D) in combination to counteract the neurodegenerative disorders induced by CuSO4 intake in experimental rats. Neurodegeneration (Alzheimer-like) was induced in twenty-four male Wistar albino rats by CuSO4 supplement to drinking water (10 mg/L) for 14 weeks. AD rats were divided into four groups: untreated AD group (Cu-AD) and three treated AD groups; orally treated for 4 weeks with either DPZ (10 mg/kg/day), Vit D (500 IU/kg/day), or DPZ + Vit D starting from the 10th week of CuSO4 intake. Another six rats were used as normal control (NC) group. The hippocampal tissue content of ß-amyloid precursor protein cleaving enzyme 1 (BACE1), phosphorylated Tau (p-tau), clusterin (CLU), tumor necrosis factor-α (TNF-α), caspase-9 (CAS-9), Bax, and Bcl-2 and the cortical content of acetylcholine (Ach), acetylcholinesterase (AChE), total antioxidant capacity (TAC), and malondialdehyde (MDA) were measured. Cognitive function tests (Y-maze) and histopathology studies (hematoxylin and eosin and Congo red stains) and immunohistochemistry for neurofilament. Vit D supplementation alleviated CuSO4-induced memory deficits including significant reduction hippocampal BACE1, p-tau, CLU, CAS-9, Bax, and TNF-α and cortical AChE and MDA. Vit D remarkably increased cortical Ach, TAC, and hippocampal Bcl-2. It also improved neurobehavioral and histological abnormalities. The effects attained by Vit D treatment were better than those attained by DPZ. Furthermore, Vit D boosted the therapeutic potential of DPZ in almost all AD associated behavioral and pathological changes. Vit D is suggested as a potential therapy to retard neurodegeneration.

Sources and levels of copper affect liver copper profile, intestinal morphology and cecal microbiota population of broiler chickens fed wheat-soybean meal diets.

Super dosing copper (Cu) has long been used as an alternative to antibiotic growth-promoters in broiler chickens' diet to improve gut health. This study was designed to compare nutritional and growth-promoting levels of Cu hydroxychloride (CH) with CuSO4 on gut health bio-markers and liver mineral profile of broiler chickens. Ross 308 chicks (n = 864) were randomly assigned to eight treatments, as basal diet containing no supplemental Cu; the basal diet with 15 or 200 mg/kg Cu as CuSO4; or 15, 50, 100, 150 or 200 mg/kg Cu from CH. The highest liver Cu content was observed in birds fed the diets with 200 mg/kg CuSO4 (P < 0.01). Serum FITC-d concentration as the leaky gut marker, and liver malondialdehyde concentration were not affected. Copper level or source had no effect on cecal short chain fatty acid and the mRNA expression of five jejunal genes involved in gut integrity. Negative linear responses of Cu were observed on Lactobacillus (P = 0.032), Bacteroides (P = 0.033), and Enterobacteriaceae (P = 0.028) counts. The jejunal villus height increased in birds fed CH at 200 and 100 mg/kg (P < 0.05). Increasing Cu levels, linearly and quadratically (P < 0.001), increased Cu excretion.

Comparative toxicity of nanoparticulate and ionic copper following dietary exposure to common carp (Cyprinus carpio).

Copper is an essential element for the normal growth and survival of all organisms including fish. However, its excessive presence in the environment can cause bioaccumulation and aquatic toxicology. The aim of the present study was to compare the dietary toxicity effects of two different Cu compounds, copper oxide nanoparticles (CuO-NPs) and ionic copper (CuSO4) in juvenile common carp, Cyprinus carpio. To prepare experimental diets, two nominal concentrations of 100 and 1000 mg Cu kg-1 diet were added to a basal diet. Carp (n = 450, average initial weight of 35.94 ± 5.35 g) were fed on the Cu-supplemented diets and basal diets for two 21-day courses as dietary exposure and recovery periods, respectively. The growth performance, survival rate and blood biochemical indices as well as copper accumulation in target organs of fish were investigated at the end of each exposure period. The results showed that the weight gain (WG) of carp significantly decreased coincident with increasing concentration of the both dietary Cu forms (P = 0.00). Both Cu sources at concentrations of 100 mg kg-1 diet decreased the survival rate of fish (P = 0.003), likely due to more feed intake and thus increased copper toxicity. The both forms of dietary Cu at two different concentrations significantly decreased the plasma glutamate oxaloacetate transaminase (GOT) level compared to the control group (P = 0.008). Fish exposed to diets containing Cu sources except 100 mg Cu kg-1 of CuO-NPs showed the lower glutamate pyruvate transaminase (GPT) activity in comparison to the control (P = 0.00). The plasma sodium level in1000 mg CuO-NPs kg-1 diet was significantly lower than the control (P = 0.001). The plasma potassium level increased in the all Cu-supplemented groups except 100 mg kg-1 of CuO-NPs after the dietary exposure period (P = 0.035). The copper accumulation was dose-dependent in all target organs. In 100 mg Cu kg-1 dietary groups, the liver showed the highest Cu accumulation (P = 0.00), while in 1000 mg Cu kg-1 dietary groups, the highest Cu content was observed in the intestine (P = 0.00). The results demonstrated the enhanced toxicological responses in fish after 21 days of dietary exposure, but the levels of most of biochemical indices and tissues Cu content decreased or returned to the control values after the recovery period.

Effects of Organic Copper on Growth Performance and Oxidative Stress in Mice.

Copper (Cu) has been used as a feed additive for many years. However, high Cu amounts can cause oxidative stress and adversely affect animal performance. Such negative effects may depend on the amounts and forms of Cu. In the present study, the effects of inorganic Cu (CuSO4) and organic Cu (chelate-Cu) present in mice feed on daily growth rate and Cu deposition in the liver, kidneys, spleen, brain, and serum were assessed in addition to the oxidative stress levels in the liver and brain. Organic Cu at a concentration of 15 mg/kg significantly enhanced daily growth rate in mice, whereas Cu deposition in the livers was significantly lower than that in the inorganic Cu group. Glutathione peroxidase activity in the liver of the mice fed with organic Cu significantly improved, whereas malondialdehyde levels in the brain and liver were significantly lower than that in the inorganic Cu group. The different effects of organic Cu and inorganic Cu provide key evidence supporting the use of organic Cu in animal feeds.

Effects of soybean oil and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls.

The study evaluated the effects of soybean oil (SO) and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls. Eight Holstein rumen-cannulated bulls (14 ± 0.2 months of age and 326 ± 8.9 kg of body weight) were allocated into a replicated 4 × 4 Latin square design in a 2 × 2 factorial arrangement with factors being 0 or 40 g/kg dietary dry matter (DM) of SO and 0 or 7.68 mg/kg DM of Cu from copper sulphate (CS). The basal diet contained per kg DM 500 g of corn silage, 500 g of concentrate, 28 g of ether extract (EE) and 7.5 mg of Cu. The SO × CS interaction was significant (p < 0.05) for ruminal propionate proportion and acetate to propionate ratio. Dietary SO addition increased (p < 0.05) intake and total tract digestibility of EE but did not affect average daily gain (ADG) of bulls. Dietary CS addition did not affect nutrient intake but increased (p < 0.05) ADG and total tract digestibility of DM, organic matter, crude protein and neutral detergent fibre. Ruminal pH was not affected by treatments. Dietary SO addition did not affect ruminal total volatile fatty acids (VFA) concentration, decreased (p < 0.05) acetate proportion and ammonia N and increased (p < 0.05) propionate proportion. Dietary CS addition did not affect ammonia N, increased (p < 0.05) total VFA concentration and acetate proportion and decreased (p < 0.05) propionate proportion. Acetate to propionate ratio decreased (p < 0.05) with SO addition and increased (p < 0.05) with CS addition. Dietary SO addition decreased (p < 0.05) activity of carboxymethyl cellulase, cellobiase and xylanase as well as population of fungi, protozoa, methanogens, Ruminococcus albus and R. flavefaciens but increased (p < 0.05) α-amylase activity and population of Prevotella ruminicola and Ruminobacter amylophilus. Dietary CS addition increased (p < 0.05) activity of cellulolytic enzyme and protease as well as population of total bacteria, fungi, protozoa, methanogens, primary cellulolytic and proteolytic bacteria. Microbial protein synthesis was unchanged with SO addition but increased (p < 0.05) with CS addition. The results indicated that the addition of CS promoted nutrient digestion and ruminal fermentation by stimulating microbial growth and enzyme activity but did not relieve the negative effects of SO addition on ruminal fermentation in dairy bulls.

Copper and zinc sources and levels of zinc inclusion influence growth performance, tissue trace mineral content, and carcass yield of broiler chickens.

A 35-d experiment was conducted in broilers to study the effect of supplementation of sulfate or hydroxychloride forms of Zn and Cu at 2 supplemental Zn levels on growth performance, meat yield, and tissue levels of Zn. On day 0, 900 male Ross 308 broiler chicks (45 ± 1.10 g) were allocated to 4 treatments in a randomized complete block design and 2 × 2 factorial arrangement of treatments. The factors were 2 sources (sulfate or hydroxychloride) of Zn and Cu and 2 levels (low or high) of Zn. The Zn sources were zinc sulfate monohydrate (ZSM) or hydroxychloride Zn. Copper sources were copper (II) sulfate pentahydrate or hydroxychloride Cu. Each of the 4 treatments had 15 replicates and 15 birds per replicate. Birds were weighed on days 0, 21, and 35 for growth performance. On day 35, left tibia bone, liver, and blood were collected from 4 randomly selected birds per pen. In addition, 7 birds per pen were used for carcass evaluation. There was no significant source × level interaction on any of the growth performance response. Broiler chickens receiving hydroxychloride Zn and Cu had greater (P < 0.05) gain: feed, whereas broiler chickens receiving lower Zn level had greater (P < 0.01) weight gain. There was no source × level interaction on meat yield. Broiler chickens receiving hydroxychloride Zn and Cu had greater (P < 0.05) % breast yield than those receiving sulfate Zn and Cu. Higher level of Zn, irrespective of source, produced greater (P < 0.01) tibia and plasma Zn levels, whereas liver Cu was greater (P < 0.05) in broiler chickens receiving hydroxychloride Zn and Cu. It was concluded that hydroxychloride Zn and Cu were more efficacious than sulfate Zn and Cu in promoting growth performance and enhancing meat yield in the current study.

Copper requirements of broiler breeder hens.

One-hundred-twenty Cobb 500 hens, 20 wk of age, were randomly allocated into individual cages with the objective of estimating Cu requirements. After being fed a Cu deficient diet for 4 wk, hens were fed diets with graded increments of supplemental Cu (0.0; 3.5; 7.0; 10.5; 14; and 17.5 ppm) from Cu sulfate (CuSO4 5H2O), totaling 2.67; 5.82; 9.38; 12.92; 16.83; and 20.19 ppm analyzed Cu in feeds for 20 weeks. Estimations of Cu requirements were done using exponential asymptotic (EA), broken line quadratic (BLQ), and quadratic polynomial (QP) models. Obtained Cu requirements for hen d egg production and total settable eggs per hen were 6.2, 7.3, and 12.9 ppm and 8.1, 9.0, and 13.4 ppm, respectively, using EA, BLQ, and QP models. The QP model was the only one having a fit for total eggs per hen with 13.1 ppm Cu as a requirement. Hemoglobin, hematocrit, and serum Cu from hens had requirements estimated as 13.9, 11.3, and 18.5, ppm; 14.6, 13.0, and 19.0 ppm; and 16.2, 14.6, and 14.2 ppm, respectively, for EA, BLQ, and QP models. Hatching chick hemoglobin was not affected by dietary Cu, whereas requirements estimated for hatching chick hematocrit and body weight and length were 10.2, 12.3, and 13.3 ppm using EA, BLQ, and QP models; and 6.8 and 7.1 ppm, and 12.9 and 13.9 ppm Cu using EA and BLQ models, respectively. Maximum responses for egg weight, yolk Cu content, and eggshell membrane thickness were 14.9, 12.7, and 15.1 ppm; 15.0, 16.3, and 15.7 ppm; and 7.3, 7.8, and 14.0 ppm Cu, respectively, for EA, BLQ, and QP models. Yolk and albumen percentage were adjusted only with the QP model and had requirements estimated at 11.0 ppm and 11.3 ppm, respectively, whereas eggshell mammillary layer was maximized with 10.6, 10.1, and 14.4 ppm Cu using EA, BLQ, and QP models, respectively. The average of all Cu requirement estimates obtained in the present study was 12.5 ppm Cu.

Feeding inhibition in Corbicula fluminea (O.F. Muller, 1774) as an effect criterion to pollutant exposure: Perspectives for ecotoxicity screening and refinement of chemical control.

Bivalves are commonly used in biomonitoring programs to track pollutants. Several features, including its filter-feeding abilities, cumulatively argue in favour of the use of the Asian clam (Corbicula fluminea) as a biosentinel and an ecotoxicological model. Filtration in bivalves is very sensitive to external stimuli and its control is dictated by regulation of the opening/closure of the valves, which may be used as an avoidance defence against contaminants. Here, we investigate the filter-feeding behaviour of the Asian clam as an endpoint for assessing exposure to pollutants, driven by two complementary goals: (i) to generate relevant and sensitive toxicological information based on the ability of C. fluminea to clear an algal suspension, using the invasive species as a surrogate for native bivalves; (ii) to gain insight on the potential of exploring this integrative response in the refinement of chemical control methods for this pest. Clearance rates and proportion of algae removed were measured using a simple and reproducible protocol. Despite some variation across individuals and size classes, 50-90% of food particles were generally removed within 60-120 min by clams larger than 20 mm. Removal of algae was sensitive to an array of model contaminants with biocide potential, including fertilizers, pesticides, metals and salts: eight out of nine tested substances were detected at the µg l-1 or mg l-1 range and triggered valve closure, decreasing filter-feeding in a concentration-dependent manner. For most toxicants, a good agreement between mortality (96 h - LC50 within the range 0.4-5500 mg l-1) and feeding (2 h - IC50 within the range 0.005-2317 mg l-1) was observed, demonstrating that a 120-min assay can be used as a protective surrogate of acute toxicity. However, copper sulphate was very strongly avoided by the clams (IC50 = 5.3 µg l-1); on the contrary, dichlorvos (an organophosphate insecticide) did not cause feeding depression, either by being undetected by the clams' chemosensors and/or by interfering with the valve closure mechanism. Such an assay has a large potential as a simple screening tool for industry, environmental agencies and managers. The ability of dichlorvos to bypass the Asian clam's avoidance strategy puts it in the spotlight as a potential agent to be used alone or combined with others in eradication programs of this biofouler in closed or semi-closed industrial settings.

Comparative study on the impact of copper sulphate and copper nitrate on the detoxification mechanisms in Typha latifolia.

The present study focused on cupric sulphate and cupric nitrate uptake in Typha latifolia and the impact of these copper species on the plant's detoxification capacity. When the plants were exposed to 10, 50 and 100 µM cupric sulphate or cupric nitrate, copper accumulation in T. latifolia roots and shoots increased with rising concentration of the salts. Shoot to root ratios differed significantly depending on the form of copper supplementation, e.g. if it was added as cupric (II) sulphate or cupric (II) nitrate. After incubation with 100 µM of cupric sulphate, up to 450 mg Cu/kg fresh weight (FW) was accumulated, whereas the same concentration of cupric nitrate resulted in accumulation of 580 mg/kg FW. Furthermore, significant differences in the activity of some antioxidative enzymes in Typha roots compared to the shoots, which are essential in the plant's reaction to cope with metal stress, were observed. The activity of peroxidase (POX) in roots was increased at intermediate concentrations (10 and 50 µM) of CuSO4, whereas it was inhibited at the same Cu(NO3)2 concentrations. Ascorbate peroxidase (APOX) and dehydroascorbate reductase (DHAR) increased their enzyme activity intensely, which may be an indication for copper toxicity in T. latifolia plants. Besides, fluorodifen conjugation by glutathione S-transferases (GSTs) was increased up to sixfold, especially in roots.

Effect of mineral source and mannan oligosaccharide supplements on zinc and copper digestibility in growing pigs.

This study aimed to compare the effects of organic (proteinate) and inorganic (sulphate) copper (Cu) and zinc (Zn) supplements, in presence or absence of a mannan oligosaccharide (MOS) supplement, on mineral solubility and digestibility in pigs. Twenty-eight barrows (25 ± 4 kg) assigned randomly to four treatment groups were fed a corn-wheat-soya bean meal diet with 10 mg/kg of Cu and 100 mg/kg of Zn supplied as organic or inorganic supplement, and supplemented or not with 0.1% MOS. After an adaptation period, total faeces and urine were collected for a period of 6-7 days. Pigs were then euthanatised and digesta from ileum and caecum were collected. Apparent digestibility was calculated in ileum and caecum using titanium dioxide. The organic mineral supplement improved total (faecal) digestibility and retained/ingested ratio of Cu (p < 0.05) while reducing apparent digestibility of Zn in the ileum (p < 0.05) without effect on total digestibility of Zn. Solubilities of Cu and Zn in liquid fraction of ileum and caecum were not affected by mineral sources. Although MOS supplement increased Cu solubility in the ileum (p < 0.05), it had no effect on digestibility of Zn and Cu in ileum, caecum and faeces, retained/ingested ratio of Zn and Cu, or pH and volatile fatty acid concentration in ileal and caecal digesta. In conclusion, organic mineral supplement improved total digestibility and retained/ingested ratio of Cu in pigs but this cannot be attributed to its solubility in ileal and caecal digesta. The MOS supplement did not interfere with digestibility or dietary utilisation of Zn and Cu in pigs fed above the Zn and Cu requirements.

Effect of copper on the expression of IGF-1 from chondrocytes in newborn piglets in vitro.

The experiment was performed to evaluate the influence of copper supplementation on insulin-like growth factor-1 (IGF-1) mRNA expression in chondrocytes of newborn pigs. Chondrocytes were isolated and cultured in media containing 15% fetal calf serum supplemented with 0, 15.6, 31.2, and 62.5 µmol/L copper in 90-mm culture plate. After 0, 12, 24, and 48 h, total RNA was isolated from chondrocytes. Then, IGF-1 mRNA expression was determined by semiquantitative RT-PCR. The results showed that the expression of IGF-1 mRNA, adjusted for ß-actin expression, was increased in the culture media added to 15.6, 31.2, and 62.5 µmol/L copper, respectively. In the present experiment, the optimal copper concentration and optimal culture time for the expression of IGF-1 mRNA were 31.2 µmol/L and 48 h, respectively.

Enhanced in vitro regeneration and change in photosynthetic pigments, biomass and proline content in Withania somnifera L. (Dunal) induced by copper and zinc ions.

In the present study the effect of inorganic nutrients (CuSO4 & ZnSO4) on morphogenic and biochemical responses from nodal explants in Withania somnifera L. was investigated. Incorporation of either Copper sulphate (25-200 µM) or Zinc sulphate (50-500 µM) in the optimized Murashige and Skoog (MS) medium highly influenced the shoot bud formation and subsequent elongation, which induced maximum percentage (95%) regeneration, number (61.7 ± 0.25) of shoots with shoot length (5.46 ± 0.16 cm) on CuSO4 (100 µM) and maximum percentage regeneration (100%), number of shoots (66.1 ± 0.96) with shoot length (6.24 ± 0.21 cm) on ZnSO4 (300 µM) after 12 weeks of culture. Healthy growing in vitro microshoots rooted efficiently on ½ MS medium supplemented with NAA (0.5 µM), which induced (16.2 ± 0.12) roots with root length (3.30 ± 0.12 cm) after 4 weeks. Pigment content increased with increasing concentration of Cu and Zn and the maximum Chl. a (0.47), (0.41); Chl. b (0.52), (0.42); total Chl. (0.99), (0.83) and Carotenoid (0.16), (0.16) mg/g FW contents in regenerants were found on CuSO4 (100 µM) and ZnSO4 (300 µM), respectively. Maximum proline content (0.17), (0.16) µg/g FW was observed on high concentrations of CuSO4 (200 µM) and ZnSO4 (500 µM) respectively, in the basal medium. Regenerated plantlets were acclimatized successfully in soilrite with a survival rate of 95%. No morphological variations were detected among the micropropagated plants when compared with seedling raised plants of the same age.

Effects of dietary copper supplementation on production performance and plasma biochemical parameters in broiler chickens.

A study was conducted to estimate the effect of copper (Cu) supplementation on growth performance and biochemical profiles of blood and meat in broiler chickens. A total of 240 d-old broiler chicks (Vencobb-100) were randomly divided into 12 groups, each of 20 chicks (4 treatments x 3 replicates). The basal diet (T1) contained 215 g kg⁻¹ crude protein (CP), 12·76 MJ kg⁻¹ ME, 32 g kg⁻¹ total calcium and 5 g kg⁻¹ total phosphorus. T2, T3 and T4 were formulated to contain an additional 75, 150 and 250 mg Cu kg⁻¹ diet, respectively. Copper sulphate pentahydrate (CuSO4, 5H2O) was used as the source of Cu. Significant reductions in plasma total cholesterol and triglyceride, and an elevated concentration of HDL-cholesterol, were observed in the chickens fed with 250 mg Cu kg⁻¹ (T4) of feed at the 3rd and 6th week of the experiment. Total cholesterol in meat decreased significantly in the birds fed with dietary Cu at 250 mg kg⁻¹ (T4) of feed. Growth performance was measured in terms of live weight gain, cumulative feed intake and feed conversion ratio at the end of d 21 and d 42 of the experiment, and the result was found to be commercially beneficial for the chickens receiving 150 mg Cu kg⁻¹ (T3) of diet. The concentration of Cu in breast muscle and liver increased significantly at the end of experiment. From this study it can be concluded that supplementation with dietary Cu may be beneficial for production performance and plasma biochemical characteristics of broiler chickens.

Selection of fecal enterococci exhibiting tcrB-mediated copper resistance in pigs fed diets supplemented with copper.

Copper, as copper sulfate, is increasingly used as an alternative to in-feed antibiotics for growth promotion in weaned piglets. Acquired copper resistance, conferred by a plasmid-borne, transferable copper resistance (tcrB) gene, has been reported in Enterococcus faecium and E. faecalis. A longitudinal field study was undertaken to determine the relationship between copper supplementation and the prevalence of tcrB-positive enterococci in piglets. The study was done with weaned piglets, housed in 10 pens with 6 piglets per pen, fed diets supplemented with a normal (16.5 ppm; control) or an elevated (125 ppm) level of copper. Fecal samples were randomly collected from three piglets per pen on days 0, 14, 28, and 42 and plated on M-Enterococcus agar, and three enterococcal isolates were obtained from each sample. The overall prevalence of tcrB-positive enterococci was 21.1% (38/180) in piglets fed elevated copper and 2.8% (5/180) in the control. Among the 43 tcrB-positive isolates, 35 were E. faecium and 8 were E. faecalis. The mean MICs of copper for tcrB-negative and tcrB-positive enterococci were 6.2 and 22.2 mM, respectively. The restriction digestion of the genomic DNA of E. faecium or E. faecalis with S1 nuclease yielded a band of ∼194-kbp size to which both tcrB and the erm(B) gene probes hybridized. A conjugation assay demonstrated cotransfer of tcrB and erm(B) genes between E. faecium and E. faecalis strains. The higher prevalence of tcrB-positive enterococci in piglets fed elevated copper compared to that in piglets fed normal copper suggests that supplementation of copper in swine diets selected for resistance.

Effect of type of muscle and Cu supplementation on trace element concentrations in cattle meat.

Considering that meat is an important source of metals exposure to humans it is important to explore trace element concentrations in different types of muscles. Because of the demonstrated effect of Cu-supplementation on mineral status, the influence of Cu-supplementation was also evaluated. Samples of four different muscles (diaphragm, cardiac, semitendinous and pectoral, n=120) from beef calves receiving typical commercial diets Cu-supplemented (15 mg Cu(2)SO(4)/kg DM) and non-supplemented were taken and acid digested. The levels of non-essential (As, Cd, Hg, Pb and Sn) and essential (Co, Cr, Fe, Mn, Mo, Ni, Se and Zn) elements were analyzed by ICP-MS. The statistical analyzes included two way Anova, post hoc DHS Tukey and Spearman correlations. The most active and less fat containing muscles showed in general the highest essential and the lowest non-essential trace element accumulation. As and Hg muscular residues are indicative of animal exposure, however, in situations of an adequate mineral status, essential trace element concentrations in muscle are irrespective of the mineral status of the animal and could be possibly related to their own particular muscular metabolism. Cu-supplementation significantly reduced As but caused a significant decrease of Se, which could have significance for the animal's health.

Growth response, blood characteristics and copper accumulation in organs of broilers fed on diets supplemented with organic and inorganic dietary copper sources.

1. A 56-d experiment was conducted to study the comparative influence of organic and inorganic dietary copper (Cu) sources on growth, blood characteristics and copper accumulation in organs of broilers. 2. A total of 480 Arbor-Acre unsexed broilers were fed on diets containing copper sulphate (CuSO(4)) or copper proteinate (Cu Pro) at concentrations of 50, 100 or 150 mg/kg of Cu supplementation. The birds were given a broiler starter diet from 1-28 d and a broiler finisher diet from 29-56 d which contained 30·8 mg/kg and 41·1 mg/kg basal copper concentration respectively. Growth performance, blood characteristics and Cu accumulation in organs of the broilers were measured. 3. At 28 d, Cu Pro-fed birds had improved feed conversion ratio compared with CuSO(4). At 56 d, birds fed on Cu Pro diets had significantly greater body weight than CuSO(4)-fed birds. Birds fed on CuSO(4) supplemented diets had significantly better feed conversion efficiency. Feed consumptions for the two Cu sources were not significantly different. At no stage did the concentration of added Cu affect the productive traits measured. 4. Cu Pro supplementation increased haemoglobin concentration but reduced plasma triglyceride and plasma cholesterol. Plasma cholesterol decreased as Cu concentration increased. 5. There was a greater accumulation of Cu in the blood, heart, lung, liver and bone of broilers fed on Cu Pro than in those receiving CuSO(4). The liver Cu concentration increased as dietary Cu concentration increased. 6. Cu Pro was more effective in promoting growth and reducing blood cholesterol, and was more bio-available in the organs of broilers.

Changes in composition a metabolism of caecal microbiota in rats fed diets supplemented with copper-loaded chitosan nanoparticles.

The study was conducted to investigate the effect of copper-loaded chitosan nanoparticles on the composition and metabolism of the caecal microbiota in rats. Forty male Sprague-Dawley (SD) rats (average body weight of 82 ± 5 g) were randomly assigned to five groups (n = 8). The dietary treatments were: (i) basal diet, (ii) basal diet + 80 mg/kg BW CuSO(4), (iii) basal diet + 80 mg/kg BW chitosan (CS-I), (iv) basal diet + 80 mg/kg BW copper-loaded chitosan nanoparticles (CSN-I) and (v) basal diet + 160 mg/kg BW copper-loaded chitosan nanoparticles (CSN-II). The trial lasted 21 days. The results showed that compared with control, Average day gain (ADG) of group CSN-I and CSN-II increased (p < 0.05). No significant difference was found in CuSO(4) or CS-I-treated groups (p > 0.05). There were no effects of these treatments on average day feed intake (ADFI) of rats (p > 0.05). The counts of Bifidobacteria and Lactobacillus in group CSN-II were higher than those of the control group (p < 0.05), while the counts of total aerobes, total anaerobes, Salmonella, Clostridium and coliform were lower than those of the control (p < 0.05). The activity of ß-glucuronidase in group CSN-I or CSN-II was significantly depressed (p < 0.05), while the activities of α-galactosidase and ß-galactosidase were enhanced significantly (p < 0.05). The pH of the caecum digesta and the concentration of propionate and butyrate in group CSN-I and CSN-II were higher than those of the control group (p < 0.05). No significant difference was found in these parameters in CuSO(4) or CS-I-treated groups (p > 0.05). The results indicate that the microbiota and environment of caecum are beneficially changed by the administration of copper-loaded CSN.

A newly isolated Paecilomyces sp. WSH-L07 for laccase production: isolation, identification, and production enhancement by complex inducement.

Laccase can catalyze the oxidation of a wide range of organic and inorganic substrates. In this study, an easily detectable method was employed for screening laccase-producing microorganisms by using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) as laccase-secretion indicator. A novel laccase-producing strain was isolated and identified as Paecilomyces sp. WSH-L07 according to the morphological characteristics and the comparison of internal transcribed spacer (ITS) ribosomal DNA (rDNA) gene sequences. In further investigation, the production of laccase by Paecilomyces sp. WSH-L07 was greatly enhanced by the nontoxic inducers of copper sulphate and methylene blue. Under the induction of 50 microM copper sulphate and 20 microM methylene blue, the maximum laccase production was obtained. When these inducers were added into cultivation medium at 24 h and 12 h, respectively, an increment of about 100 times of laccase activity compared with that of in inducer-free medium and about two times of that of in single copper-supplemented medium was observed. Compared with other Paecilomyces species, Paecilomyces sp. WSH-L07 exhibit the better laccase-producing characteristics with an activity of 1,650 U/l on the eighth day, suggesting its potential ability for industrial application.