Tribasic Copper Chloride Research Articles

355 Determining the bioavailability of copper carbonate (Emerald-C) and tribasic copper chloride relative to copper sulfate in feedlot steers under moderate antagonistic pressure

Abstract Copper (Cu) from Cu sulfate (CuSO4) may be more susceptible to antagonism when fed in cattle diets high in sulfur and molybdenum, but other Cu supplements may be better able to avoid this. The objective of this study was to determine the bioavailability of Cu carbonate (CuCO3; Emerald-C, Old Bridge Minerals, Old Bridge, NJ) and tribasic Cu chloride (TBCC; Intellibond-C, Selko, Indianapolis, IN) relative to CuSO4 when fed to steers supplemented with 2 mg Mo/kg DM and 0.1% S. Angus crossbred steers [n = 84; body weight (BW) = 282 ± 19.3 kg] were enrolled (n = 12 per treatment) in a 96-d study and fed a corn-silage based diet with one of seven Cu supplementation strategies: no supplemental Cu (CON), 5 or 10 mg supplemental Cu/kg DM from CuSO4, 5 or 10 mg supplemental Cu/kg DM from TBCC, or 5 or 10 mg supplemental Cu/kg DM from CuCO3. Steers were housed in pens equipped with GrowSafe feed bunks (GrowSafe Systems Ltd., Airdire, AB, Canada) with six steers per pen. The GLM procedure of SAS 9.4 was used to determine the bioavailability of CuCO3 and TBCC relative to CuSO4 using both liver and plasma Cu. The MIXED procedure of SAS was used to analyze liver Cu, plasma Cu, and growth performance data. Contrasts compared CON vs. 5 mg/kg, CON vs. 10 mg/kg, 5 mg/kg vs. 10 mg/kg, CuCO3 vs. CuSO4, TBCC vs. CuSO4, and CuCO3 vs. TBCC. The bioavailability of CuCO3 relative to CuSO4 based on liver and plasma Cu was 89 (P < 0.01) and 133% (P = 0.21), respectively. The bioavailability of TBCC relative to CuSO4 based on liver and plasma Cu was 88 (P < 0.01) and 100% (P = 0.82), respectively. Over the 96-d trial, liver Cu decreased 66% in CON, from 74 to 21 mg Cu/kg DM. Steers receiving CuCO3 had greater plasma Cu concentrations than those receiving TBCC on d 56 (P = 0.05) but were similar by d 96 (P = 0.39). Copper supplemented steers had greater dry matter intake (DMI) than CON (P = 0.01). Steers supplemented with CuCO3 had greater DMI than those receiving TBCC (P = 0.05). Steers receiving TBCC tended to have greater gain:feed than those receiving CuSO4 (P = 0.08), but gain:feed did not differ between CON and supplemented treatments (P ≥ 0.6). In conclusion, CuCO3 and TBCC may be less available to steers under moderate antagonistic pressure based on liver Cu, but Emerald-C CuCO3 may result in greater Cu in circulation. More research is needed to evaluate the impact of feeding CuCO3 on systemic Cu metabolism and how factors such as initial Cu status impact relative bioavailability results.

199 Phytosorb improves feed efficiency in pigs of two different commercial Duroc sire lines in late finishing compared with tri-basic copper chloride

Abstract The objective of this experiment was to determine whether Phytosorb could replace tribasic copper chloride in grow-finish rations with equal or better performance, and evaluate this in two Duroc sire lines. A total of 1,080 growing pigs [initial body weight (BW) = 35.3 ± 1.05 kg) from DNA female with 2 different sire lines (A and B) were randomly assigned to 2 dietary treatments in a randomized complete block design. There were 27 pigs in each pen and 10 replicate pens per treatment. The 2 dietary treatments were 1) standard diet with an additional 150 ppm of Cu supplementation (CON); 2) standard diet without copper supplementation + 0.45 kg of Phytosorb (PHY) a source of natural flavors and soy lecithin. Body weight and feed disappearance were recorded by pen on d 0, 21, 35, 49, 64, and 77, and were used to calculate average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F). After d 77, an equal number of pigs from each pen were weighed and marketed over 5 individual cuts. Data were used to calculate all parameters and were analyzed using the GLM procedure in Minitab with Fisher’s t-test determining differences between treatments. The initial model included treatment, genetics (sire lines), and the interaction between treatment and genetics, however, the interaction was excluded in the final model due to being non-significant on the response criteria. Results indicated that ADG was greater (P < 0.05) and ADFI tended (P < 0.10) to be greater in pigs fed CON compared with pigs fed PHY from d 0 to 64 Table 1). G:F was greater (+5.9%, P < 0.05) from d 64 to 77, and G:F tended (+3.8%, P < 0.10) to be greater from d 64 to market weight (MKT; approximately 104 to - 134 kg BW) in pigs fed PHY compared with pigs fed CON. For the overall finishing phase (approx. 35.5 to 134 kg BW), ADG tended to be greater (P < 0.10) in pigs fed CON compared with pigs fed PHY but no differences in ADFI and G:F were observed. For genetics, there were G:F differences (P < 0.05, respectively) between sire line A and B from d 0 to 64, d 64 to MKT, and d 77 to MKT and overall finishing phase. Phytosorb was not able to replace copper in early finishing; the response during this phase was what we would expect copper to do. In late finishing, however, these results demonstrate that Phytosorb may give equal ADG, with an improved G:F ratio leading to an improved cost per kg of gain as compared with feeding tribasic copper chloride. This feeding strategy would support environmental sustainability by reducing the usage of copper in finishing diets.

Investigations into Increased Incidence of Severe Gizzard Erosions and Ulcerations in U.S. Commercial Broilers.

During a series of pathology surveys in four production complexes of a U.S. broiler integrator, the technical services veterinarians of an animal health company noted a high incidence of severe gizzard erosions and ulcerations (GEU), prompting further clinical investigation and a battery trial. No growth-promoting antibiotics or ionophore coccidiostats were used during the period of these surveys. All used tribasic copper chloride (TBCC) at ≤120 ppm added copper in broiler rations. Clostridium perfringens was isolated from 83% and 67% of gizzard lesions cultured in two complexes, and cecal C. perfringens most probable number determinations were higher in severely affected than in mildly affected or unaffected birds. Histopathology revealed both acellular koilin fusion defects characteristic of copper toxicity, as well as inflammatory cell infiltrates. Intralesional bacilli suggestive of C. perfringens were noted in 78% of affected flocks examined. Species E Aviadenovirus was isolated from one bird in one complex, and that bird had a single intranuclear inclusion body; no other flocks had Adenoviruses isolated or detected on PCR, nor any inclusion bodies. Other viruses detected were thought to be incidental. A pilot study using feed with supplemental copper from TBCC or copper sulfate and challenge with one of the isolated C. perfringens strains reproduced the lesions. A battery study was conducted with an unchallenged negative control group fed a diet with 16 ppm added copper, a group fed the control diet and orally challenged with 108 organisms of a field strain of C. perfringens at 21 and 22 days, and a group treated with the same diet containing 250 ppm added copper from TBCC and orally challenged with C. perfringens. Birds were necropsied at 23 and 28 days. All challenged groups developed lesions, with those receiving both TBCC and C. perfringens having significantly higher gross and histopathological lesion scores than the unchallenged negative controls. Lesions were qualitatively similar to those in the field and contained suspected C. perfringens bacilli. Because the levels of TBCC used in the commercial birds and in the battery trial generally have been considered safe, and because C. perfringens is usually regarded as a pathogen of the lower GI tract, the possible association of these two agents with GEU is a novel observation and warrants further investigation.

269 Copper Methionine Hydroxy Analogue Chelate as a Reduce and Replace Cu Strategy in Growing Pigs

Abstract Research has shown that minerals supplied in the methionine hydroxy analogue chelate (MHAC; MINTREX Trace Minerals [Novus International Inc., St. Charles, MO]) form are more available than inorganic trace minerals. Research in weaned pigs demonstrated improved pig performance when Cu was provided in the Cu-MHAC form compared with inorganic Cu sources. In the growing pig it may be desirable to reduce the amount of total Cu fed to minimize Cu excretion while maintaining animal performance. Data from 8 studies, 4 studies per Cu source comparison, was used to compare 50 mg/kg of Cu as Cu-MHAC with either 125 mg/kg Cu as CuSO4 or 150 mg/kg Cu supplied in the tri-basic copper chloride (TBCC) form. There were 44 pens of 23 pigs per pen for each Cu source in the Cu-MHAC vs TBCC comparison and 39 pens of 22 pigs per pen for each Cu source for the Cu-MHAC vs. CuSO4 comparison. Cu-MHAC was compared with CuSO4 in separate trials from those comparing Cu-MHAC vs TBCC. Trials were conducted across all seasons. Pigs had previously received either CuSO4 or TBCC during the nursery phase. Data were analyzed with the PROC MIXED procedure of SAS using trial as a random effect. Outlier analysis was conducted for each parameter with any observation greater than 3 standard deviations from the treatment mean removed. Pigs in the Cu-MHAC vs CuSO4 comparisons averaged 29.3 + 2.18 kg of BW while those used in the Cu-MHAC vs TBCC comparisons averaged 31.0 + 2.33 kg of BW. Average study duration was 105 days. Average market weight sold was 0.45 kg/pig (P = 0.52) and 0.59 kg/pig (P = 0.46) less for Cu-MHAC than CuSO4 and TBCC pigs respectively. Adjusted feed conversion was 4 points lower (P = 0.12) and 2 points lower (P = 0.25) for the Cu-MHAC vs CuSO4 and TBCC pigs respectively. Removal percentage during the studies ranged from 3.63% to 4.91% with the Cu-MHAC pigs having 0.6 percentage points less removals than the CuSO4 (P = 0.52) and 0.7 percentage points less removals than the TBCC (P = 0.38) treatment. The fewer removals resulted in the Cu-MHAC treatment having 3 points decreased feed conversion (P = 0.25) than TBCC pigs and 6 points decreased feed conversion (P = 0.12) than CuSO4 pigs on a pen basis. The combined Cu-MHAC analysis vs combined inorganic Cu source analysis showed that Cu-MHAC pigs had 4 points less feed conversion on both an adjusted (P = 0.06) and closeout (P = 0.06) basis than pigs fed inorganic Cu. The cumulative removal rate was 0.7 percentage points less for the Cu-MHAC pigs (P = 0.29). These results indicate supplying Cu as 50 mg/kg Cu-MHAC can reduce feed conversion compared with 125 mg/kg Cu as CuSO4 and 150 mg/kg Cu as TBCC in growing-finishing diets.

268 Copper Methionine Hydroxy Analogue Chelate in a Reduce and Replace Copper Strategy Improves Nutrient Availability in Growing Pigs

Abstract Research has shown that methionine hydroxy analogue chelated minerals [MHAC; MINTREX Trace Minerals (Novus International Inc., St. Charles, MO)] are more available than inorganic trace minerals. In growing pigs, 50 mg/kg of Cu as Cu-MHAC has been shown to reduce feed conversion compared with feeding 125 mg/kg Cu as CuSO4 and 150 mg/kg Cu as tri-basic copper chloride (TBCC). A metabolism study was conducted using 2 groups of 10 barrows (average BW of 33.2 + 3.03 kg) fed either 50 mg/kg Cu as Cu-MHAC or 150 mg/kg Cu as TBCC. There was a 14-d acclimation period followed by 3-d of total fecal and 5-d of total urine collection. Corn-soybean meal diets with 15% distillers dried grains and < 1% fat as soy oil were formulated to 0.98% SID lysine with 0.65% Ca and 0.49% P. Phytase was included at 1,500 FTU/kg. A common basal trace mineral premix supplying 120 mg/kg Zn, 10 mg/kg Cu and 20 mg/kg Mn in the SO4 form was used. Diets were provided in meal form twice daily at 5% of body weight per day. The total trial period was 21-d with BW measured at d 0 and 21. Feed disappearance was recorded throughout. Data were analyzed using the PROC MIXED procedure of SAS with treatment and initial BW as fixed effects and group as a random effect. Pigs fed Cu-MHAC gained 816 g/d compared with 787 g/d for those fed TBCC (P = 0.55). Feed conversion was 2.07 for TBCC pigs and 1.98 for Cu-MHAC pigs (P = 0.48). Dry matter digestibility was 90.9% and 89.9% (P = 0.18) for Cu-MHAC and TBCC pigs respectively. Dry matter fecal output was 9% less for Cu-MHAC pigs (P = 0.24). Cu retention percent was not different between Cu sources (64.4% Cu-MHAC vs 62.4% TBCC; P = 0.63). Cu excretion was less for Cu-MHAC pigs (41.3 mg/d Cu-MHAC vs. 103.9 mg/d TBCC; P < 0.001) as a result of reduced Cu intake. Cu-MHAC increased Zn (P < 0.05) and Mn (P < 0.01) retention by 8.9 and 11.1 percentage points, respectively. Iron absorption (P = 0.86) and retention (P = 0.81) were not affected by Cu source. Calcium absorption and retention were 3.8 percentage points greater (P = 0.12) for Cu-MHAC pigs. Absorption and retention of P was 3.4% (P = 0.25) and 2.7% greater (P = 0.49) for pigs fed Cu-MHAC. Cu-MHAC resulted in 1.9% greater N absorption (P = 0.06) and 8.7% greater N retention (P = 0.06%) than when 150 mg/kg Cu in the TBCC form was fed. These results indicate a reduce and replace Cu strategy with Cu-MHAC can reduce Cu excretion by 60% and improved overall mineral and N retention with the exception of Fe. These nutrient availability improvements may contribute to the improved feed conversion observed in performance trials with Cu-MHAC vs inorganic sources.

Long-term oral tribasic copper chloride exposure impedes cognitive function and disrupts mitochondrial metabolism by inhibiting mitophagy in rats

Copper (Cu) is an essential micronutrient element that commonly acted as a feed additive and antimicrobial in agricultural production. Tribasic copper chloride (TBCC) is a relatively new dietary Cu source, and its exposure directly or indirectly affects the safety of animals and ecological environment, thus posing a potential risk to human health. Cu overexposure would produce toxic reactive oxygen species (ROS) that may have toxic effects on the host, but the mechanism of neurotoxicity remains unclear. Herein, to explore the effects of long-term TBCC-induced neurotoxicity, 150 male Sprague-Dawley rats were randomly allocated and treated with different doses of TBCC, and the cortical and hippocampus tissues were harvested at 0, 6, and 12 weeks after treatment. Morris Water Maze (MWM) test showed that excessive intake of TBCC could induce cognitive dysfunction in rats. Moreover, after treatment with 160 mg/kg Cu (276 mg/kg TBCC) for 12 weeks, pathological changes were observed in the cortex and hippocampus, and the number of Nissl bodies decreased significantly in the hippocampus. Additionally, mitochondrial structure was significantly altered and neuronal mitochondrial fusion/fission equilibrium was disrupted in 80 mg/kg and 160 mg/kg Cu groups at 12 weeks. With an increase in TBCC dose and treatment time, the number of mitophagosomes and the expression of mitophagy-related genes were significantly decreased after initially increasing. Furthermore, metformin (Met) and 3-methyladenine (3-MA) were used to regulate the level of mitophagy to further explore the mechanism of Cu-induced nerve cell injury in vitro., and it found that mitophagy activator (Met) would increase mitochondrial fission, while mitophagy inhibitors (3-MA) would aggravate mitochondrial metabolic disorders by promoting mitochondrial fusion and inhibiting mitochondrial division. These results indicate that long-term oral TBCC could impede cognitive function and disrupts mitochondrial metabolism by inhibiting mitophagy, providing an insightful perspective on the neurotoxicity of dietary TBCC.

A new tribasic copper chloride adsorbent's synthesis, characterization and analysis of its regeneration using a Fenton-like procedure

A new tribasic copper chloride adsorbent's synthesis, characterization and analysis of its regeneration using a Fenton-like procedure

Assessment of cecal microbiota modulation from piglet dietary supplementation with copper

BackgroundSwine production expanded in the last decades. Efforts have been made to improve meat production and to understand its relationship to pig gut microbiota. Copper (Cu) is a usual supplement to growth performance in animal production. Here, two performance studies were conducted to investigate the effects of three different sources of Cu on the microbiota of piglets. A total of 256 weaned piglets were randomly allocated into 4 treatments (10 replicates per treatment of 4 piglets per pen in Trial 1 and 8 replicates of 3 piglets per pen in Trial 2). Treatments included a control group (fed 10 mg/kg of Cu from CuSO4), a group fed at 160 mg/kg of Copper (II) sulfate (CuSO4) or tri-basic copper chloride (TBCC), and a group fed with Cu methionine hydroxy analogue chelated (Cu-MHAC) at 150, 80, and 50 mg/kg in Phases 1 (24–35 d), 2 (36–49 d), and 3 (50–70 d), respectively. At 70 d, the cecum luminal contents from one pig per pen were collected and polled for 16 S rRNA sequencing (V3/V4 regions). Parameters were analyzed in a completely randomized block design, in which each experiment was considered as a block.ResultsA total of 1337 Operational Taxonomic Units (OTUs) were identified. Dominance and Simpson ecological metrics were statistically different between control and treated groups (P < 0.10) showing that different Cu sources altered the gut microbiota composition with the proliferation of some bacteria that improve gut health. A high abundance of Prevotella was observed in all treatments while other genera were enriched and differentially modulated, according to the Cu source and dosage. The supplementation with Cu-MHAC can modify a group of bacteria involved in feed efficiency (FE) and short chain fatty acids (SCFA) production (Clostridium XIVa, Desulfovibrio, and Megasphera). These bacteria are also important players in the activation of ghrelin and growth hormones that were previously reported to correlate with Cu-MHAC supplementation.ConclusionsThese results indicated that some genera seem to be directly affected by the Cu source offered to the animals. TBCC and Cu-MHAC (even in low doses) can promote healthy modifications in the gut bacterial composition, being a promising source of supplementation for piglets.

Effects of bis-chelated copper in growth performance and gut health in broiler chickens subject to coccidiosis vaccination or coccidia challenge.

Copper (Cu) is widely used at high levels as growth promoter in poultry, the alternative source of Cu to replace the high level of inorganic Cu at poultry farm remains to be determined. Three floor pen experiments were conducted to evaluate the effects of Cu methionine hydroxy-analogue chelate (Cu-MHAC, MINTREX®Cu, Novus International, Inc.) on growth performance and gut health in broilers in comparison to CuSO4 and/or tribasic copper chloride (TBCC). There were 3 treatments in experiment#1 (0, 30 and 75ppm Cu-MHAC) and experiment#2 (15 and 30ppm Cu-MHAC, and 125 ppm CuSO4), and 4 treatments in experiment #3 (15 and 30ppm Cu-MHAC, 125ppm CuSO4 and 125ppm TBCC) with nine replicates pens of 10-13 birds in each treatment. The levels of other minerals were equal among all treatments within each experiment. All birds were orally gavaged with a coccidiosis vaccine at 1x recommended dose on d0 in experiment#1 and #2 and 10x recommended dose on d15 in experiment #3. Data were analyzed by one-way ANOVA, means were separated by Fisher's protected LSD test. A p ≤ 0.05 was considered statistically different. In experiment #1, 30 and 75ppm Cu-MHAC improved FCR during grower phase, increased jejunal villus height and reduced jejunal crypt depth, 30 ppm Cu-MHAC increased cecal Lactobacillus spp. abundance in 41days broilers. In experiment #2, compared to CuSO4, 15ppm Cu-MHAC increased cumulative performance index in 28days broilers, 15 and/or 30ppm Cu-MHAC improved gut morphometry, and 30ppm Cu-MHAC reduced the abundance of E. coli and Enterobacteriaceae in cecum in 43days broilers. In experiment #3, 15ppm and 30ppm Cu-MHAC improved FCR vs. CuSO4 during starter phase, reduced the percentage of E. coli of total bacteria vs. TBCC, 30ppm Cu-MHAC increased the percentages of Lactobacillus acidophilus, Lactobacillus spp. and Clostridium cluster XIVa of total bacteria vs. both CuSO4 and TBCC in the cecum of 27days broilers. In summary, low doses of Cu-MHAC had comparable growth performance to high dose of TBCC and CuSO4 while improving gut microflora and gut morphometry in broilers subject to coccidiosis vaccination or coccidia challenge, indicating that low doses of bis-chelated Cu could be used as a complimentary strategy to improve animal gut health.

Effect of methionine and trace minerals (zinc, copper and manganese) supplementation on growth performance of broilers subjected to Eimeria challenge.

Coccidiosis is a major intestinal challenge that causes economic loss to the broiler industry. Two battery cage studies were conducted to evaluate the effect of trace minerals, source and dose of methionine on growth performance and gut health of broilers subjected to Eimeria challenge. Experiment #1 consisted of 9 treatments of 2 × 2 × 2 factorial design + 1 arrangement with main factors of methionine (Met) sources (DL-Met vs. 2-hydroxy-4-(methylthio)-butanoic acid (HMTBa)), total sulfur amino acid (TSAA) levels (high vs. low; ±5% of recommended level), and sources of trace minerals (TM) Zn:Cu:Mn in the form Inorganic trace minerals (ITM) in sulfates (80:20:100ppm) vs. mineral methionine hydroxy-analogue bis-chelate (MMHAC, 40:10:50ppm), each with 8 pens of 10 birds. Experiment #2 consisted of 2 treatments--ITM [ZnSO4:tribasic copper chloride (TBCC):MnSO4 110:125:120ppm] and MMHAC (Zn:Cu:Mn, 40:30:40ppm), each with 36 pens of 10 birds. All birds except for treatment 9 in experiment #1 were orally gavaged with 1x, 4x and 16x recommended dose of coccidiosis vaccine on d0, d7 and d14, respectively. Data were subjected to one-way and/or three-way ANOVA, and means were separated by Fisher's protected LSD test with significance at p ≤ 0.05. In experiment #1, factorial analysis revealed the main effects of TSAA level and TM, but not Met source. High TSAA level improved body weight and cumulative feed intake at 14, 20, and 27d. MMHAC improved body weight at 14, and 27d; feed intake at 14, 20, and 27d; and cumulative FCR at 27d vs. sulfates. One-way ANOVA analysis showed that birds fed MMHAC and high levels of TSAA regardless of Met source had similar body weight as unchallenged birds on d27. In experiment #2, MMHAC improved body weight and cumulative FCR, and reduced jejunal IL-17A gene expression on d28. In summary, in broilers subjected to Eimeria challenge, supplementation of the reduced levels of bis-chelated trace minerals MMHAC improved growth performance compared to high levels of ITM (sulfates or TBCC), which might partially result from better immune response, high levels of TSAA improved growth performance, Met source had no effect. Supplementation of both bis-chelated trace minerals MMHAC and high levels of TSAA could overcome the growth performance challenge issue due to coccidiosis.

Effects of Copper Sulfate and Encapsulated Copper Addition on In Vitro Rumen Fermentation and Methane Production

Copper is a microelement crucial for the proper functioning of animals’ metabolic processes. The function of copper in rumen fermentation processes and methanogenesis is not well analyzed. The aim of the study was to evaluate the different types of copper supplement, their rumen decomposition and effect on in vitro ruminal fermentation as well as methanogenesis. Two different copper additives were used in the experiment: CS—copper sulfate (CuSO4 × 5 H2O)—and EC—encapsulated copper (tribasic copper chloride and copper sulfate enclosed within a polysaccharide polymer coating). A total mixed ration without copper additive was used as a control (C). In vitro rumen fermentation was conducted, and fermentation profile, gas production and methanogenesis were evaluated. After 24 h of fermentation, the amount of copper in the rumen fluid was significantly higher in the CS group. EC was protected against rumen degradation to a greater extent. The type of used copper supplement affects rumen fermentation. However, the effect on methanogenesis is ambiguous. CS supplement increases rumen gas production but does not affect methanogenesis. The obtained results suggest that the EC supplement may reduce the risk of low-fat milk and may improve the economic indicators of milk production. An in vivo experiment is necessary to compare the obtained in vitro results with animal productivity.

Electron-deficient Cu site catalyzed acetylene hydrochlorination

Rational design of catalytic sites to activate the CC bond is of paramount importance to advance acetylene hydrochlorination. Herein, Cu sites with electron-rich and electron-deficient states were constructed by controlling the impregnation solutions. The π electrons flowing from acetylene to Cu site are facilitated over the electron-deficient Cu sites, achieving high activation of CC bond. The contradiction between the increased activation of acetylene required for enhanced catalytic activity and the resistance of Cu site to reduction by acetylene required for maintaining catalytic stability can be balanced by establishing strong interactions of Cu site with pyrrolic-N species. The catalytic activity displays a volcano shape scaling relationship as a function of Cu particle size. Tribasic copper chloride is concomitantly generated with the construction of electron-deficient Cu sites. The H–Cl bond of HCl can be activated over the tribasic copper chloride, accelerating the surface reaction of vinyl chloride production. This strategy of inducing electron deficiency provides new insight into the rational design of catalysts for the synthesis of vinyl chloride with a high catalytic performance.

Development of an Immunoassay for the Detection of Copper Residues in Pork Tissues.

The presence of high concentrations of copper (Cu) residues in pork is highly concerning and therefore, this study was designed to develop a high-throughput immunoassay for the detection of such residues in edible pork tissues. The Cu content in the pork samples after digestion with HNO3 and H2O2 was measured using a monoclonal antibody (mAb) against a Cu (II)–ethylenediaminetetraacetic acid (EDTA) complex. The resulting solution was neutralized using NaOH at pH 7 and the free metal ions in the solution were chelated with EDTA for the immunoassay detection. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method was developed for Cu ion analysis. The half maximal inhibitory concentration of the mAb against Cu (II)–EDTA was 5.36 ng/mL, the linear detection range varied between 1.30 and 27.0 ng/mL, the limit of detection (LOD) was 0.43 μg/kg, and the limit of quantification (LOQ) was 1.42 μg/kg. The performances of the immunoassay were evaluated using fortified pig serum, liver, and pork samples and had a recovery rate of 94.53–102.24%. Importantly, the proposed immunoassay was compared with inductively coupled plasma mass spectroscopy (ICP-MS) to measure its performance. The detection correlation coefficients of the three types of samples (serum, pork, and liver) were 0.967, 0.976, and 0.983, respectively. Thirty pork samples and six pig liver samples were collected from local markets and Cu was detected with the proposed ic-ELISA. The Cu content was found to be 37.31~85.36 μg/kg in pork samples and 1.04–1.9 mg/kg in liver samples. Furthermore, we detected the Cu content in pigs with feed supplemented with tribasic copper chloride (TBCC) and copper sulfate (CS) (60, 110, and 210 mg/kg in feed). There was no significant difference in Cu accumulation in pork tissues between the TBCC and CS groups, while a remarkable Cu accumulation was found for the CS group in liver at 210 mg/kg, representing more than a two-fold higher level than seen in the TBCC group. Therefore, the proposed immunoassay was found to be robust and sensitive for the detection of Cu, providing a cost effective and practical tool for its detection in food and other complicated samples.

Chronic tribasic copper chloride exposure induces rat liver damage by disrupting the mitophagy and apoptosis pathways

Despite the fact that copper (Cu) is a vital micronutrient to maintain body function, high doses of Cu through environmental exposure damage various organs, especially the liver, which is the main metabolic organ. To investigate the influence of long-term Cu-induced toxicity on mitophagy and apoptosis in rat liver, 96 seven-month-old male Sprague-Dawley rats were fed TBCC for 24 weeks. The results revealed that exposure to high Cu concentrations could promote oxidative stress liver injury by increasing the hepatic function index (ALT, AST and ALP) and MDA content, while reducing the activity of antioxidant enzymes (T-SOD, GSH-Px and CAT) related to oxidative stress. Consistent with histopathological observations, proper dietary Cu (15–60 mg/kg) could improve antioxidant stress levels and induce a dose-dependent increase in the mRNA expression of mitophagy-related genes, whereas a high Cu concentration (120 mg/kg) could cause severe liver impairment and ultrastructural changes and a reduction in mitophagosomes, accompanied by downregulation of Atg5, Beclin1, Pink1, Parkin, NIX, P62 and LC3B. The expression of apoptosis-related genes (Bax, Bax/Bcl-2, Caspase3, Cytc and p53) and proteins (Caspase3 and p53) was upregulated with the addition of dietary Cu. The results demonstrated that an appropriate dose of TBCC could improve liver function by promoting mitophagy and Cu enzymes that play antioxidative roles, while the accumulation of excess Cu could induce liver lesions by enhancing apoptosis and inhibiting mitophagy pathways.

LOW DOSE OF COPPER IS MORE EFFECTIVE ON INTESTINAL MORPHOLOGY IN WEANED PIGS

The present experiment was carried out to determine the relation between pharmacological levels of copper (Cu) (125 and 200 mg/kg) and source (cooper sulfate (CuSO 4 ) and tribasic copper chloride (TBCC)) on growth performance, diarrhea incidence and intestinal morphology of piglets after weaning. At 21 days of age, 96 weaned piglets were sorted by body weight (BW, 6.14 ± 0.269 kg) and randomly assigned to 4 treatments according to a block design of a 2 x 2 factorial arrangement: level (125 and 200 mg/kg) and source (CuSO 4 or TBCC). Growth performance was evaluated by measuring BW, average daily feed intake (ADFI), average daily gain (ADG) and feed conversion ratio (FCR) from 21 to 63 days of age. Fecal score was evaluated daily and at 63 days of age tissue samples were collected from the duodenum, jejunum and ileum for intestinal morphology analysis. At this moment, it was evaluated villus height (VH), crypt depth (CD) and villus height to crypt depth ratio (VH:CD ratio). There was no difference between level and source (P&gt;0.05) for any growth performance. Piglets fed 200 mg/kg TBCC had the lowest diarrhea frequency (P&lt;0.05) in overall nursery period than the other treatments during the whole experimental period. Supplementation with 125 mg/kg increased VH (P&lt;0.05) and VH:CD ratio (P&lt;0.05) in the duodenum when compared to 200 mg/kg, regardless of the source. Based on our results, we can conclude that 125 mg/kg of Cu improve the VH and VH:CD ratio in the duodenum and may show growth performance results similar to 200 mg/kg of Cu.

PSIII-32 Interactive effects of copper sources and superdose phytase on growth performance, nutrient digestibility and tissue mineral concentrations in nursery pigs

Abstract This study investigated the interactive effects of copper (Cu) sources and superdose phytase on growth performance, nutrient digestibility and tissue mineral concentrations in nursery pigs. A total of 288 weaning barrows (PIC 337 × 1050, Hendersonville, TN; BW = 6.06 ± 0.99 kg), blocked by BW, were randomly allotted to 1 of 6 dietary treatments, with 12 pens/treatments and 4 pigs/pen. Six dietary treatments were arranged in 3 × 2 factorial design, with 3 Cu sources [125 ppm Cu from copper sulfate (CuSO4), tribasic copper chloride (TBCC) or copper methionine hydroxy analogue chelate (Cu-MHAC, MINTREX® Cu, Novus International, Inc.), respectively] and 2 phytase levels [0 or 1500 FTU/kg from phytase PHYTAVERSE® G10 (Novus International, Inc.)]. There was an interaction (P = 0.05) between Cu sources and superdose phytase on ADG during d 0–41, with the improvement by superdose phytase being 25.84, 33.96 and 40.85% when the Cu sources were CuSO4, TBCC and Cu-MHAC, respectively. Superdose phytase significantly (P &amp;lt; 0.01) reduced F:G during d 0 to 41. Cu-MHAC led to greater ATTD of neutral and acid detergent fiber (NDF, ADF, respectively; P = 0.01) than CuSO4 and TBCC, respectively. Cu-MHAC led to greater (P = 0.03) ATTD of P than CuSO4, with TBCC intermediate. There was no interaction (P &amp;gt; 0.10) between Cu sources and superdose phytase in terms of nutrient digestibility and tissue mineral concentrations. Cu source did not (P &amp;gt; 0.10) affect tissue mineral concentrations; however, superdose phytase increased (P ≤ 0.05) bone Ca and P concentrations. In conclusion, Cu-MHAC may improve the efficacy of superdose phytase in terms of growth performance, most likely by improving phytate P hydrolysis and therefore providing more P for absorption. Superdose phytase is effective in improving growth performance of nursery pigs, mainly driven by increased Ca and P absorption and utilization.

Determining the relationship between varying inclusions of Bacillus lichenformis and tribasic copper chloride on 42-day-old Ross 708 male broiler performance

Summary Probiotics, such as Bacillus licheniformis (BL), and some trace minerals, such as copper (supplied via tribasic copper chloride; [TBCC]), have been explored singularly in the literature for use in antibiotic-free poultry production. However, these additives are often applied in combination, without much peer-reviewed research conducted on their interactive effects. Therefore, the objective was to examine the individual and interactive effects of supplementing commercially available BL (yes or no) and TBCC (0, 125, or 250 ppm) within an all-vegetable diet on day 0–42 male broiler performance. Interactions between BL x TBCC were established for day 18–30 BW gain (BWG) and day 0–30 feed intake (FI)/bird. For BWG, 125-ppm TBCC maximized gain, whereas including BL + TBCC decreased gain. For day 0–30 FI, feeding TBCC alone increased the FI, whereas inclusions of BL + TBCC decreased the FI; from day 0 to 30, regardless of the FI, the feed conversion ratio was not affected. For day 30–42 BWG, broilers receiving 250-ppm TBCC had the highest BWG, whereas broilers fed all other TBCC inclusions performed similarly. On day 42, 5 birds/treatment were randomly selected for cecal tonsil sampling to determine the presence of Salmonella or Escherichia coli. Although Salmonella was not detected, birds fed diets containing TBCC had less E. coli as compared with birds fed diets without TBCC; BL had no effect on E. coli. Overall, no significant overall performance differences were found. However, the observed intermittent BL × TBCC interactions demonstrate their combined use may negatively affect performance, warranting further research to ensure they and other combined antibiotic alternatives do not have antagonistic effects.

Optimal dietary copper requirements and relative bioavailability for weanling pigs fed either copper proteinate or tribasic copper chloride

BackgroundThe objective of this study was to determine the effects of supplementing Cu on growth performance, Cu metabolism and Cu-related enzyme activities of weanling pigs fed diets with two different Cu sources, and to estimate optimal Cu requirements and relative bioavailability from these two sources for pigs.MethodsWeanling pigs were allocated to 14 treatments arranged factorially, including 6 added Cu levels (5, 10, 20, 40, 80, 160 mg/kg), and 2 mineral sources (tribasic Cu chloride, TBCC and copper proteinate, CuPro), as well as one negative control (0 mg/kg added Cu level) and one maximum allowed level treatment (200 mg/kg TBCC) for the entire 38-d experiment. Growth performance, mineral status and enzyme activities were measured at the end of this study.ResultsIncreasing levels of Cu showed linear and quadratic responses (P < 0.01) for final BW, ADG and FCR regardless of the sources. Supplementation with TBCC (> 80 mg/kg) and CuPro (> 20 mg/kg) significantly decreased (P < 0.05) diarrhea incidence of weanling pigs. There were linear and quadratic increases (P < 0.01) in bile, hepatic, and intestinal Cu concentrations, fecal Cu contents, and plasma enzyme activities (alkaline phosphatase, ceruloplasmin, Cu, Zn-Superoxide dismutase (Cu/Zn SOD), and glutathione peroxidase), whereas plasma malondialdehyde decreased (P < 0.01) linearly and quadratically as dietary Cu level increased. Similarly, pigs fed CuPro absorbed and retained more Cu and excreted less Cu than those fed TBCC when supplemented 80 mg/kg and above. Optimal dietary Cu requirements for pigs from 28 to 66 d of age estimated based on fitted broken-line models (P < 0.05) of bile Cu, plasma Cu/Zn SOD and growth performance were 93–140 mg/kg from TBCC, and 63–98 mg/kg from CuPro accordingly. According to slope ratios from multiple linear regression, the bioavailability value of CuPro relative to TBCC (100%) was 156–263% (P < 0.01).ConclusionThe findings indicated that Cu recommendation from current NRC (5–6 mg/kg) was not sufficient to meet the high requirement of weanling pigs. Cu from CuPro was significantly more bioavailable to weanling pigs than TBCC in stimulating growth and enzyme activities, decreasing diarrhea frequency and fecal Cu contents to the environment.

Pre-treatment of petroleum refinery wastewater by coagulation and flocculation using mixed coagulant: Optimization of process parameters using response surface methodology (RSM)

The present work reports the efficacy of three coagulants CuSO4, FeCl3 and CuSO4+FeCl3 (both salt mixed in 1:1 ratio (v/v)) for treatment of wastewater from effluent treatment plant of petroleum refinery using coagulation and flocculation process. The independent parameters such as pH and dosage of coagulants were optimized using response surface methodology with central composite design technique considering final pH, reduction in COD, turbidity, TDS and color as dependent variables. The statistical fitness of developed model was checked using ANOVA. A second order quadratic model (R2> 95 %) was best fitted to develop the relation between independent and dependent variables. Charge neutralization, sweep flocculation, adsorption and formation of tribasic copper chloride (TBCC) were mainly responsible for coagulation and flocculation processes. Results showed that mixed coagulant CuSO4+FeCl3 delivered better result than CuSO4 and FeCl3 individually. CuSO4+FeCl3 along with adsorption and sweep flocculation, forms tribasic copper chloride (TBCC) as a proceeding intermediate in the pH range 5−7. The TBCC formation enhanced the flocculation by destabilizing colloidal and suspended particles due to its catalytic, decolorizing and octahedral structural properties. Maximum reduction in COD (76.77 %), turbidity (89.47 %), TDS (94.16 %) and color (95.29 %) was observed at pH 7.12 and 0.20 g/L dosage of CuSO4+FeCl3 coagulant. The final pH of solution at optimum condition for all three coagulants was under the disposal limits. Further, the flocs formed are characterized using XPS, SEM and EDX analyses to reveal the formation of TBCC as intermediate during the process.

Effects of Copper Sources and Levels on Lipid Profiles, Immune Parameters, Antioxidant Defenses, and Trace Element Residues in Broilers.

The study was conducted to investigate the effects of copper sources and levels on lipid profiles, immune parameters, antioxidant defenses, and trace element contents of meat and liver in Arbor Acres broilers. A total of 504 male broilers were randomly divided into 7 groups with 6 replicates per group and 12 broilers per replicate. The experiment was used in a 3 × 2 + 1 factorial experiment design; broilers in the control group were fed a basal diet, and broilers in the other six groups were fed basal diets supplemented with 3 sources (copper sulfate, tribasic copper chloride, and copper methionate) and 2 levels (10 and 20mg/kg). The results showed that the levels of cholesterol and low-density lipoprotein cholesterol in broilers were significantly decreased with the increase of dietary copper level (P <0.05). Serum IL-6 and IgA contents, ceruloplasmin and GSH-Px activities, and liver copper contents of broilers increased significantly with dietary copper levels (P <0.05). Compared with the control group, dietary copper supplementation significantly decreased serum cholesterol (P <0.05) and significantly increased serum IL-6, ceruloplasmin, SOD, GSH-Px, and liver copper (P <0.05). Dietary supplementation of basic copper chloride and copper methionate significantly decreased low-density lipoprotein cholesterol content and liver iron content (P < 0.05). In conclusion, dietary copper supplementation can effectively reduce serum cholesterol content and improve immune and antioxidant functions in broilers. Adding 20mg/kg copper to broiler diet can increase the copper content in the liver, but it will not affect the copper content in the chicken.