Plasma Ceruloplasmin Research Articles

Relationship of blood and seminal plasma ceruloplasmin, copper, iron and cadmium concentrations with sperm quality in Merino rams

The aim of the current study was to investigate the concentrations of ceruloplasmin, copper, iron, zinc and cadmium concentrations in blood serum and seminal plasma obtained from Merino rams. In addition, their relationship with sperm parameters, fertility rate and litter size were also studied. Blood and ejaculate samples (6 replicates) were taken in October from 19 Merino rams, aged between 18 and 24 months. Ceruloplasmin, copper, iron, zinc and cadmium in blood serum and seminal plasma were determined. Sperm parameters including volume, mass motility, motility, concentration, Hos-test, viability, abnormal sperm and acrosome abnormality in semen, fertility rate and litter size were also evaluated. Highly positive correlation was found between blood ceruloplasmin and blood copper concentrations (r=0.812, p<0.001), whereas negative correlation were determined between these parameters in seminal plasma (r=−0.195, p<0.05). Seminal plasma copper concentration was positively correlated with seminal plasma cadmium (r=0.206, p<0.05) and seminal plasma iron (r=0,305, p<0.01) concentrations. Negative correlation was determined between blood ceruloplasmin level and acrosomal defect (r=−0.443, p<0.05). Seminal plasma ceruloplasmin level was positively correlated with volume (r=0.255, p<0.01) and negatively correlated with abnormal sperm (r=−0.186, p=0.058) and acrosome abnormality (r=−0.213, p<0.05). Seminal plasma iron concentration was positively correlated with other abnormality (r=0.257, p<0.01). Seminal plasma cadmium concentration was positively correlated with sperm abnormality (r=0.207, p=0.052) and other abnormality (r=0.262, p<0.05) and negatively correlated with fertility rate (r=−0.449, p=0.054). Blood cadmium concentration was negatively correlated with litter size (r=−0.579, p<0.01). In conclusion, blood and seminal plasma ceruloplasmin may be suggested to have positive influence regardless of copper with its antioxidant property whereas iron and cadmium have negative influence on sperm parameters and fertility in Merino rams.

Rosmarinic acid modulates the antioxidant status and protects pancreatic tissues from glucolipotoxicity mediated oxidative stress in high-fat diet: streptozotocin-induced diabetic rats.

Persistent hyperglycemia and elevated levels of free fatty acids (FFA) contribute to oxidative stress, a proximate cause for the onset and progression of diabetes and its complications. The present study was hypothesized to evaluate the anti-diabetic potential of Rosmarinic acid (RA) during high-fat diet (HFD)-streptozotocin (STZ)-induced type 2 Diabetes (T2D) in wistar albino rats. Oral administration of RA (100 mg/kg b.w) significantly (p < 0.05) increased the insulin sensitivity index (ISI0,120), while the levels of blood glucose, HbA1c, advanced glycation end products (AGE), TNF-α, IL-1β, IL 6, NO, p-JNK, P38 MAPK and NF-κB were significantly reduced, with a concomitant elevation in the plasma insulin levels in diabetic rats. Furthermore, RA treatment significantly (p < 0.05) reduced the levels of triglycerides, FFA and cholesterol in serum, and reduced the levels of lipid peroxides, AOPP's and protein carbonyls in the plasma and pancreas of diabetic rats. The diminished activities of pancreatic superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) and the decreased levels of plasma ceruloplasmin, vitamin C, vitamin E and reduced glutathione (GSH) in diabetic rats were also significantly (p < 0.05) recovered upon RA treatment denoting its antioxidant potential which was confirmed by Nrf-2, hemeoxyenase (HO-1) levels. Histological, ultrastructural and immunohistochemical data demonstrate that oral administration of RA protects pancreatic β-cells from oxidative niche in HFD-STZ-induced experimental diabetes. Our findings suggest that the oral treatment with RA alleviates pancreatic β-cell dysfunction and glucolipotoxicity-mediated oxidative stress during HFD-STZ-induced T2DM, perhaps through its antioxidant potential.

Radioprotective activity of neutral polysaccharides isolated from the fruiting bodies of Hohenbuehelia serotina

In this study, the radioprotective effect of neutral polysaccharides from Hohenbuehelia serotina (NTHSP) against the damages induced by 60Co-γ radiation was investigated. The results showed that NTHSP could significantly improve the activity of glutathione peroxidase (GSH-Px) and increase the contents of glutathione (GSH) and ceruloplasmin in plasma after treated with 6 Gy-radiation compared with the radiation controls (p < 0.05). Furthermore, administration with NTHSP could effectively increase the quantity of marrow DNA (p < 0.05) and reduce the rates of chromosome aberration and micronuclei (p < 0.01) in bone marrows of mice. In addition, NTHSP could markedly inhibit the expressions of Bax protein and promote the expressions of Bcl-2 protein, accordingly inhibit the releases of cytochrome c and expressions of activated Caspase-3, and therefore block the mitochondrial apoptotic pathway of splenocytes in mice induced by 60Co-γ radiation. These results suggested that NTHSP might be a natural radioprotective agent against the injuries induced by radiation.

Influence of dietary zinc and copper on apparent mineral retention and serum biochemical indicators in young male mink (Mustela vison).

An experiment was conducted in a 3 × 3 (Cu × Zn) factorial experiment based on a completely randomized design to evaluate the effects of dietary copper and zinc on apparent mineral retention and serum biochemical indicators in young male mink on a corn-fish meal based diet. Animals were fed basal diets supplemented with Cu from copper sulfate (CuSO4) and Zn from zinc sulfate (ZnSO4). Supplemental Cu levels were 0, 15, and 30 mg/kg copper, respectively, while supplemental Zn levels were 0, 150, and 300 mg/kg, respectively. A metabolism trial of 4 days was conducted during the last week of experimental feeding. Blood samples were collected via the toe clip to determine blood hematology and blood metabolites. Copper excretion, retention, and digestibility were influenced by dietary copper (P < 0.05), however, there was also a zinc-copper interaction. Copper digestibility and retention were substantially reduced when zinc was added to the low copper diet but showed little change with zinc supplementation of the high copper diet. Both plasma Cu and plasma Zn were influenced by dietary level of the respective mineral (P < 0.05). There was no influence of dietary copper on plasma zinc; however, high Zn in the diet reduced plasma Cu concentrations. There was a zinc-copper interaction for plasma Cu (P = 0.053). Cu-Zn superoxide dismutase (Cu-Zn SOD) activity tended to be influenced by dietary zinc (P = 0.065) and dietary copper (P = 0.035). Dietary copper had a significant effect on ceruloplasmin (CER) and triglyceride (TG) concentrations (P < 0.05). Our results show that moderately high Zn in the diet (Zn:Cu ratio of 40:1) significantly reduce the apparent Cu digestibility. Our results also show that moderately high Cu in the diet increases Cu retention, but not reduces Zn absorption, and moderately high Zn in the diet reduced plasma Cu concentrations and CER activity.

Peculiarity of prooxidant-antioxidant balance indicators in patients with nonalcoholic fatty liver disease who have been exposed to ionizing radiation due to the Chornobyl NPP accident

To define the features of prooxidant antioxidant balance in patients with nonalcoholic fatty liver disease who have been exposed to radiation as a result of the Chornobyl accident. using Indicators characterizing the processes of oxidative modification of macromolecules by blood lev els of compounds with isolated double bonds (CIDB), diene conjugates (DC) oxodiene conjugates (ODC), secondary products of lipid peroxiodation reacting with thiobarbituric acid, products of oxidative modification of proteins in blood serum by levels of neutral and basic 2,4 dynitrofenilhidrazones, superoxide dismutase and catalase content in erythrocytes and ceruloplasmin in plasma were determined for the diagnosis of oxidative stress. The factor of antiox idant state was calculated. In the absence of differences between the ratios between the primary and intermediate products of lipids peroxidation increasing relative content of DC and ODC in patients with nonalcoholic steatohepatitis (NASH), which suffered radiation exposure, was found compared with patients without nonalcoholic fatty liver disease (NAFLD) and NASH patients, who had no history of radiation exposure. This may be a manifestation of insignificant activation of lipid peroxidation and disturbance of LPO products transformation at different stages. In irradiated steatohepato sis (SH) patients, unlike SH patients who have not undergone radiation exposure,, the proportionalityof lipid perox idation products between the content of izopropanol phase lipid extracts of blood was violated, and Spearman cor relation coefficients between CIDB and DC and DC and ODC did not exceed the limits average values. In the group of patients with SH who suffered from radiation exposure level CIDB was slightly higher compared with patients with NASH. Regardless of whether there was a history of radiation exposure, the level of oxidative protein modification (OMB) products in NASH patients was higher compared with patients with SH, but the difference reached significant values only in the subgroup of non irradiated patients with NASH by 2.4 dinitrophenilhydrazones of neutral charac ter. Mean values ??of catalase activity in patients with NASH also exceeded the corresponding values of SH patient. The average values of the factor of antioxidant status in patients with both SH and NASH subjected to radiation exposure, were lower than in the corresponding subgroups of non irradiated patients by 29 and 21%, respectively. Existing dismetabolic disorders may contribute to the development of endogenous intoxication and deepening of pathological changes in the liver of NAFLD patients from SH to NASH.

Inclusion of anti-phospholipase A2 antibody to backgrounding diets on performance, feed efficiency, in vitro fermentation, and the acute-phase response of growing beef calves.

In Exp. 1, individual performance and daily DMI was measured on 70 crossbred weaned calves during a 70-d period using a GrowSafe system (GrowSafe Systems Ltd., Airdrie, AB, Canada) at the University of Florida North Florida Research and Education Center Feed Efficiency Facility (FEF). Calves were fed a low-concentrate (LC) growing diet, blocked by weight and sex, and then randomly assigned to pens to receive either no additional supplement (CON; n = 35) or receive a supplement of anti-phospholipase A2 antibody (aPLA2) at an inclusion rate of 0.6% of the diet DM (n = 35). After the 70-d feed efficiency (FE) trial (Phase 1), calves were loaded into a commercial livestock trailer and were driven for approximately 1,600 km during 24 h. Upon return to the FEF (Phase 2), calves were relocated to the same pens and groups and received the same diets and treatments for 28 d. Blood samples from each calf were collected on d 0, 1, 3, 5, 7, 14, 21, and 28 relative to initiation of transportation and were analyzed for determination of concentrations of plasma ceruloplasmin and haptoglobin. In Phase 1, initial BW (242.0 ± 3.7 kg; P = 0.92), BW at d 70 (313.0 ± 4.1 kg; P = 0.79), and ADG (1.01 ± 0.02 kg; P = 0.95) were similar between treatments. However, daily DMI was greater (P = 0.01) for CON (9.18 ± 0.15 kg) than aPLA2 (8.53 ± 0.15 kg). In addition, residual feed intake was greater (P = 0.002) for CON (0.389 ± 0.110 kg/d) than aPLA2 calves (-0.272 ± 0.110 kg/d). In Phase 2, after transportation, there were no differences between treatments on BW loss due to transportation shrink (26.0 ± 0.6 kg; P = 0.86), BW at d 28 (339.0 ± 4.1 kg; P = 0.72), ADG (1.28 ± 0.03 kg/d; P = 0.72), G:F (0.164 ± 0.004; P = 0.83), and concentrations of plasma haptoglobin (0.08 ± 0.02 mg/mL; P = 0.41). However, concentrations of plasma ceruloplasmin were greater (P < 0.001) for CON calves (14.3 ± 0.3 mg/dL) compared to aPLA2 calves (13.0 ± 0.3 mg/dL). In Exp. 2, the effects of aPLA2 inclusion on LC and high-concentrate (HC) substrates on in vitro fermentation parameters were assessed. Addition of aPLA2 had no effects on in vitro fermentation parameters of LC and HC substrates. In conclusion, supplementation of aPLA2 improved FE of growing beef calves when fed LC diets in Phase 1 and addition of aPLA2 had no effect on fermentation parameters of LC and HC substrates. In addition, calves supplemented with aPLA2 had reduced concentrations of plasma ceruloplasmin after 24 h of transportation.

Biomarkers of inflammation, metabolism, and oxidative stress in blood, liver, and milk reveal a better immunometabolic status in peripartal cows supplemented with Smartamine M or MetaSmart

The peripartal dairy cow experiences a state of reduced liver function coupled with increased inflammation and oxidative stress. This study evaluated the effect of supplementing basal diets with rumen-protected Met in the form of MetaSmart (MS) or Smartamine M (SM) (both from Adisseo Inc., Antony, France) during the peripartal period on blood and hepatic biomarkers of liver function, inflammation, and oxidative stress. Thirty-seven multiparous Holstein cows were fed the same basal diet from −50 to −21 d relative to expected calving [1.24Mcal/kg of dry matter (DM); no Met supplementation]. From −21 d to calving, the cows received diets (1.54Mcal/kg of DM) with no added Met (control, CON; n=13), CON plus MS (n=11), or CON plus SM (n=13). From calving through 30 d in milk (DIM), the cows received the same postpartal diet (1.75Mcal/kg of DM; CON), or CON plus MS or CON plus SM. Liver and blood samples were harvested at various time points from −21 to 21 d relative to calving. Preplanned contrasts of CON versus SM + MS during prepartum (−21 and −10 d before calving) and postpartum (7, 14, and 21 d after calving) responses were evaluated. Cows fed MS or SM compared with CON had lower overall concentrations of plasma ceruloplasmin and serum amyloid A (SAA). Compared with CON, Met-supplemented cows had greater overall plasma oxygen radical absorbance capacity. Liver concentrations of glutathione and carnitine also were greater overall with Met supplementation. Milk choline and liver phosphatidylcholine were lower overall in cows fed Met compared with controls. Liver tissue choline concentrations did not differ. Data indicate that supplemental Met enhanced de novo glutathione and carnitine synthesis in liver and, thus, increased antioxidant and β-oxidation capacity. The greater decrease of IL-6 after calving coupled with lower ceruloplasmin and SAA in Met-supplemented cows indicated a reduction in proinflammatory signaling within liver. The lower hepatic phosphatidylcholine in Met-supplemented cows might have been associated with greater assembly or export of very low density lipoproteins. Overall, biomarker analyses in blood and tissue indicate that the beneficial effect of feeding SM and MS on postpartal cow performance is due in part to a better immunometabolic status.

Effects of meloxicam administration on physiological and performance responses of transported feeder cattle.

This experiment evaluated the effects of meloxicam administration on physiological and performance responses of transported cattle during feedlot receiving. Eighty-four Angus × Hereford steers were ranked by BW on d -10 and assigned to 21 dry lot pens. From d -10 to 0, pens were fed alfalfa-grass hay ad libitum and 2.4 kg/steer daily (DM basis) of a corn-based concentrate. On d 0, pens were randomly assigned to 1) transport for 1,440 km in a livestock trailer and oral administration of meloxicam (1 mg/kg of BW) at loading (d 0), unloading (d 1), and daily from d 2 to 7 of feedlot receiving (MEL; n = 7); 2) the same transportation and treatment schedule of MEL but oral administration of lactose monohydrate (1 mg/kg of BW) instead of meloxicam (TRANS; n = 7); or 3) no transport and oral administration of lactose monohydrate (1 mg/kg of BW) concurrently with treatment administration to MEL and TRANS (CON; n = 7). Upon arrival (d 1), MEL and TRANS steers returned to their pens for a 21-d feedlot receiving with the same diet offered from d -10 to 0. Treatments were administered to steers via oral drench on d 0 and 1 or mixed daily with the concentrate from d 2 to 7. Full BW was recorded before (d -2, -1, and 0) treatment application and at the end of experiment (d 20, 21, and 22) for ADG calculation. Daily DMI was recorded from d 1 to 21. Blood samples were collected on d 0, 1, 3, 5, 7, 10, 14, and 21. During the initial 7 d of feedlot receiving, hay and total DMI were reduced (P ≤ 0.03) in TRANS vs. CON and MEL and similar between CON and MEL (P ≥ 0.26), whereas concentrate DMI did not differ (P = 0.16) among treatments. Mean ADG was reduced (P ≤ 0.03) in TRANS vs. MEL and CON but similar (P = 0.82) between MEL and CON. Moreover, TRANS had reduced G:F vs. CON (P = 0.01) and MEL (P = 0.05), whereas G:F was similar (P = 0.39) between CON and MEL. Serum NEFA concentrations were greater (P < 0.01) for TRANS and MEL vs. CON on d 1. Plasma haptoglobin concentrations were greater (P ≤ 0.03) for TRANS vs. CON and MEL on d 5 and greater (P ≤ 0.03) for CON vs. TRANS on d 10. Plasma ceruloplasmin concentrations were greater (P ≤ 0.04) for TRANS vs. CON on d 3, 5, 7, 10, and 14, greater (P ≤ 0.03) for TRANS vs. MEL on d 5 and 7, and also greater (P = 0.05) for MEL vs. CON on d 3. In conclusion, meloxicam administration to feeder steers modulated the haptoglobin and ceruloplasmin responses and prevented the performance losses caused by long-distance transportation.

Morin, a flavonoid, on lipid peroxidation and antioxidant status in experimental myocardial ischemic rats.

Myocardial infarction affects a large population in the world. Lipid peroxide metabolism plays an important role in the pathology of myocardial infarction. The present study was designed to investigate the antioxidant potential of morin, a flavonoid in isoproterenol (ISO)-induced myocardial infarction (MI), in rats. Male albino Wistar rats were pre-treated with morin (40 mg/kg), daily for a period of 30 days. After the treatment period, ISO (85 mg/kg), was subcutaneously injected in rats at an interval of 24 h for 2 days. ISO-administered rats showed elevated levels of thiobarbituric acid reactive substances (TBARS), and lipid hydro-peroxide (LOOH), in plasma and heart. Pretreatment with morin, the above changes were significantly reduced to near normal level. ISO-administered rats showed decrease in the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) in heart. In addition, decrease the levels non enzymatic antioxidants such as reduced glutathione (GSH), vitamin C and vitamin E in plasma and heart while ceruloplasmin in plasma. Pretreatment with morin, reversed these above biochemical changes towards normalcy. These findings revealed that, the morin possess antioxidant activity in experimentally induced cardiac toxicity.

Direct uptake of copper from plasma ceruloplasmin by cells that do and do not express CTR1 (996.2)

The main Cu‐binding protein of blood plasma, ceruloplasmin (Cp), has mainly been studied for its role in mediating cellular iron efflux but also has other functions, and as shown by i.v. infusion of 67Cu‐Cp, its Cu enters most major organs. To determine whether it delivers Cu directly to cells and the mechanisms involved, we incubated purified radio‐copper‐labeled and unlabeled Cp with cells and measured rates of Cu uptake and accumulation. Uptake was demonstrated for human mammary epithelial cell monolayers (PMC42 cells) with tight junctions, and with mouse embryonic fibroblasts expressing/not expressing Ctr1. Cp‐Cu was internalized. It could not be washed from the cell surface even with acidic buffers, and was found in the cytosol. Inhibitors of endocytosis did not impair uptake. Cu deficient cells exposed to Cp increased their cellular Cu contents; and holo‐Cp was converted to apo‐Cp during cell treatment, with no loss of total Cp protein. Uptake was almost the same in cells not expressing Ctr1. Excess ionic Cu(II) and Cu(I) virtually eliminated Cp‐Cu uptake. The presence of Cu(II) and Fe(III) reductase activities on the cell surface was verified. We conclude that Cp‐Cu delivers Cu to cells by a process involving cell surface reduction of Cp‐Cu(II), release of apo‐Cp, and uptake of the resulting Cu(I) by Ctr1 and an additional as yet unidentified Cu(I) transporter.Grant Funding Source: Supported by PHS Grant R15 GM100464

Plasma Ceruloplasmin, a Regulator of Nitric Oxide Activity, and Incident Cardiovascular Risk in Patients with CKD

Increased serum levels of the acute-phase reactant ceruloplasmin predict adverse clinical outcomes in the setting of acute coronary syndromes and heart failure, but their role in patients with CKD is unclear. This study investigated the relationship of ceruloplasmin with clinical outcomes in CKD, especially with regard to traditional cardiac biomarkers. Serum ceruloplasmin levels in consecutive study participants with CKD (n=654; estimated GFR<60 ml/min per 1.73 m(2)) as well as a control group of non-CKD participants matched for age and sex (n=250) were measured. Study participants were enrolled during 2001-2006 from a population of patients presenting for elective diagnostic coronary angiography and prospectively followed for 3 years (median follow-up=1095 days) to determine incident major adverse cardiac events (defined as a composite of death, nonfatal myocardial infarction, and stroke). Serum ceruloplasmin levels in CKD patients were elevated versus controls (median [interquartile range]; 25.5 [21.8-29.6] versus 22.7 [19.7-26.5] mg/dl; P<0.001) and associated with increased risk of future major adverse cardiac events (hazard ratio, 1.35; 95% confidence interval, 1.0 to 1.82; P=0.04). After adjusting for traditional risk factors, higher serum ceruloplasmin was still associated with higher risk of major adverse cardiac events at 3 years (hazard ratio, 1.61; 95% confidence interval, 1.15 to 2.25; P=0.01). In CKD patients, increased serum ceruloplasmin, a regulator of nitric oxide activity, is associated with increased risk of long-term adverse cardiovascular events, even after multivariable model adjustment for traditional clinical and biologic risk factors.

Ceruloplasmin dysfunction: a key factor in the pathophysiology of atrial fibrillation?

Ceruloplasmin dysfunction: a key factor in the pathophysiology of atrial fibrillation?

Ceruloplasmin and atrial fibrillation: evidence of causality from a population‐based Mendelian randomization study

Inflammatory diseases and inflammatory markers secreted by the liver, including C-reactive protein (CRP) and ceruloplasmin, have been associated with incident atrial fibrillation (AF). Genetic studies have not supported a causal relationship between CRP and AF, but the relationship between ceruloplasmin and AF has not been studied. The purpose of this Mendelian randomization study was to explore whether genetic polymorphisms in the gene encoding ceruloplasmin are associated with elevated ceruloplasmin levels, and whether such genetic polymorphisms are also associated with the incidence of AF. Genetic polymorphisms in the ceruloplasmin gene (CP) were genotyped in a population-based cohort study of men from southern Sweden (Malmö Preventive Project; n = 3900). Genetic polymorphisms associated with plasma ceruloplasmin concentration were also investigated for association with incident AF (n = 520) during a mean follow-up of 29 years in the same cohort. Findings were replicated in an independent case-control sample (The Malmö AF cohort; n = 2247 cases, 2208 controls). A single nucleotide polymorphism (rs11708215, minor allele frequency 0.12) located in the CP gene promoter was strongly associated with increased levels of plasma ceruloplasmin (P = 9 × 10(-10) ) and with AF in both the discovery cohort [hazard ratio 1.24 per risk allele, 95% confidence interval (CI) 1.06-1.44, P = 0.006] and the replication cohort (odds ratio 1.13, 95% CI 1.02-1.26, P = 0.02). Our findings indicate a causal role of ceruloplasmin in AF pathophysiology and suggest that ceruloplasmin might be a mediator in a specific inflammatory pathway that causally links inflammatory diseases and incidence of AF.

Wilson’s disease

Wilson s disease (WD) is a genetically determined, autosomal recessive disorder of copper metabolism. The gene ATP7B encodes a copper carrier that both transports copper from hepatocyte to bile and ceruloplasmin copper incorporation. WD may present with almost any variety of liver disease at an age ranging from 4–12 years, or with neurological and psychiatric symptoms in adolescence. Less commonly, haemolysis and/or fulminant hepatic failure may be an initial presentation. Low plasma ceruloplasmin, a positive penicillamine challenge test, and a high hepatic copper content suggest the diagnosis. Molecular methods help in diagnosing WD. If diagnosed early, it is treatable with chelators and/or zinc, and has a good prognosis. Fulminant hepatic disease has a poor outcome without transplantation.

Metabolizable protein supply modulated the acute-phase response following vaccination of beef steers

Our objective was to evaluate the effects of MP supply, through RUP supplementation, on the acute-phase response of beef steers following vaccination. On d 0, Brangus-crossbred steers (n = 24; 173 ± 31 kg; 175 ± 16 d of age) were randomly assigned to receive 1 of 3 isocaloric diets formulated to provide 85, 100, and 115% of the daily MP requirements of a beef steer gaining 0.66 kg of BW daily. Diets were limit-fed at 1.8% of BW (DM basis) and individually provided to steers once daily (0800 h) from d 0 to 29. Steers were weighed on d 0 and 29, following a 12-h period of feed and water withdrawal. On d 7, steers were vaccinated against Mannheimia haemolytica (OneShot, Pfizer), and blood samples were collected on d 0, 7, 8, 10, 14, 21, and 30. Plasma metabolites were analyzed as repeated measures using the MIXED procedure of SAS. Final BW and ADG were similar (P ≥ 0.50) among treatments (mean = 184 ± 9 kg and 0.5 ± 0.08 kg/d, respectively). Effects of time were detected (P < 0.01) for plasma concentrations of all acute-phase proteins, which peaked between d 7 to 14, returning to baseline concentrations by d 29. Treatment effects were not detected (P ≥ 0.19) for plasma concentrations of acid-soluble protein, albumin, fibrinogen, IGF-1 and serum amyloid-A. Plasma concentrations of total protein (TP) and plasma urea nitrogen (PUN) increased (P ≤ 0.05) with increasing supply of MP (87.1, 89.6, and 90.1 ± 1.09 mg TP/mL and 6.1, 8.3, and 10.3 ± 0.41 mg PUN/dL for 85, 100, and 115% MP steers, respectively). From d 10 to 29, steers provided 115% MP had less (P < 0.001) plasma concentrations of ceruloplasmin than steers fed 85 and 100% MP, which had similar plasma ceruloplasmin concentrations. On d 14, plasma concentrations of haptoglobin were greatest (P ≤ 0.06) for steers fed 115% MP, intermediate for 100% MP, and least for 85% MP (0.98, 0.71 and 0.44 ± 0.099 mg/mL, respectively). On d 10, plasma concentrations of creatinine were greater (P = 0.01) for steers fed 115 vs. 85% MP, and intermediate for steers fed 100% MP (1.63, 1.28, and 1.50 ± 0.099 mg/dL, respectively). Thus, steers provided increasing metabolizable protein had greater plasma concentrations of haptoglobin, creatinine, total protein and PUN following vaccination against M. haemolytica.

Effects of Dietary Copper on Growth, Digestive, and Brush Border Enzyme Activities and Antioxidant Defense of Hepatopancreas and Intestine for Young Grass Carp (Ctenopharyngodon idella)

To investigate the effects of dietary copper (Cu) on fish growth, digestive and absorptive enzyme activities, and antioxidant status in the hepatopancreas and intestine, young grass carp (Ctenopharyngodon idella) (282±2.8 g) were fed six diets containing 0.74 (basal diet), 2.26, 3.75, 5.25, 6.70, and 8.33 mg Cu /kg diet for 8 weeks. Results showed that percentage weight gain (PWG) and feed intake were increased with dietary Cu levels up to 3.75 mg/kg diet. In addition, the positive effects of dietary Cu at a level 3.75 or 5.25 mg/kg diet on trypsin, chymotrypsin, and lipase activities in the hepatopancreas and of Na(+), K(+)-ATPase, alkaline phosphatase, creatine kinase, and γ-glutamyl transpeptidase activities in three intestine segments produced significantly (P<0.05) better feed efficiency (FE). However, amylase activity in the hepatopancreas was decreased by dietary Cu levels up to 3.75 mg/kg diet (P<0.05). In addition, dietary Cu at 3.75 or 5.25 mg/kg diet decreased malondialdehyde and protein carbonyl content partly by significantly (P<0.05) increasing the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase, and glutathione content in the hepatopancreas and intestine. Collectively, dietary Cu improved growth and digestive and absorptive capacity and decreased lipid peroxidation and protein oxidation partly by enhancing antioxidant defense in the hepatopancreas and intestine. The dietary Cu requirement for PWG, plasma ceruloplasmin activity, and FE of young grass carp (282-688 g) were 4.78, 4.95, and 4.70 mg/kg diet, respectively.

Effects of flunixin meglumine administration on physiological and performance responses of transported feeder cattle1

The objective was to evaluate the effects of flunixin meglumine administration on physiological and performance responses of transported cattle during feedlot receiving. Forty-five Angus × Hereford steers were ranked by BW on d 0 and assigned to 1 of 3 treatments: 1) transport for 1,280 km in a commercial livestock trailer and administration of flunixin meglumine (1.1 mg/kg BW; intravenous) at loading (d 0) and unloading (d 1; FM), 2) transport for 1,280 km in a commercial livestock trailer and administration of 0.9% saline (0.022 mL/kg BW; intravenous) at loading (d 0) and unloading (d 1; TRANS), or 3) no transport and administration of 0.9% saline (0.022 mL/kg BW; intravenous) concurrently with loading (d 0) and unloading (d 1) of FM and TRANS cohorts (CON). Upon arrival and processing for treatment administration on d 1, steers within each treatment were ranked by BW and assigned to 15 feedlot pens (5 pens/treatment, 3 steers/pen). Full BW was recorded before (d -1 and 0) treatment application and at the end of experiment (d 28 and 29) for ADG calculation. Total DMI was evaluated daily from d 1 to 28. Blood samples were collected on d 0 (before treatment administration), 1 (after unloading but before treatment administration), 4, 7, 10, 14, 21, and 28. Body weight shrink from d 0 to 1 was less (P<0.01) in CON vs. FM and TRANS but similar (P=0.94) between TRANS and FM. Mean ADG was greater (P≤0.04) in CON vs. FM and TRANS but similar (P=0.69) between TRANS and FM. No treatment effects were detected on DMI, but CON had greater G:F vs. TRANS (P=0.08) and FM (P=0.02), whereas G:F was similar (P=0.68) between TRANS and FM. Mean plasma cortisol concentrations tended (P≤0.09) to be greater in TRANS vs. FM and CON but was similar (P=0.87) between CON and FM. Plasma NEFA concentrations were greater (P≤0.02) for TRANS and FM vs. CON on d 1 and greater (P≤0.04) for FM vs. TRANS and CON on d 4. Plasma ceruloplasmin concentrations were greater (P≤0.03) for TRANS vs. CON on d 1, 4, and 7, greater (P≤0.05) for TRANS vs. FM on d 4 and 7, and greater (P≤0.04) for FM vs. CON on d 1 and 4. Plasma haptoglobin concentrations were greater (P<0.01) for TRANS vs. CON and FM on d 1 and 4 and greater (P≤0.05) for FM vs. CON on d 1 and 4. In conclusion, flunixin meglumine reduced the cortisol and acute-phase protein responses elicited by road transport but did not improve receiving performance of feeder cattle.

Rest stops during road transport: Impacts on performance and acute-phase protein responses of feeder cattle1

Angus×Hereford steers (n = 42) and heifers (n = 21) were ranked by gender and BW on d 0 of the experiment and randomly assigned to 1 of 3 treatments: 1) no transport and full access to feed and water (CON); 2) continuous road transport for 1,290 km (TRANS), or 3) road transport for 1,290 km, with rest stops every 430 km (STOP; total of 2 rest stops). Treatments were applied from d 0 to 1 of the experiment. Cattle from TRANS and STOP treatments were transported in separate commercial livestock trailers, within a single 2.1×7.2 m compartment, but through the exact same route. During each rest stop, STOP cattle were unloaded and offered mixed alfalfa-grass hay and water for ad libitum consumption for 2 h. Upon arrival of STOP and TRANS on d 1, cattle were ranked by sex and BW within each treatment and assigned to 21 feedlot pens (7 pens/treatment; 2 steers and 1 heifer/pen). Full BW was recorded before (d -1 and 0) treatment application and at the end of experiment (d 28 and 29). Total DMI was evaluated daily from d 1 to 28. Blood samples were collected on d 0 (before loading of TRANS and STOP cattle), 1 (immediately after unloading of TRANS and STOP cattle), 4, 7, 10, 14, 21, and 28. Body weight shrink from d 0 to d 1 was reduced (P<0.01) in CON compared to TRANS and STOP, and reduced in STOP compared to TRANS. Mean ADG was greater (P<0.05) in CON compared to TRANS and STOP, but similar (P=0.68) between TRANS and STOP. No treatment effects were detected (P≥0.18) on hay, concentrate, and total DMI. Mean G:F was greater (P=0.05) in CON compared to STOP, tended to be greater (P=0.08) in CON compared to TRANS, and similar (P=0.85) between TRANS and STOP. Plasma cortisol concentrations were greater (P≤0.04) in TRANS compared to CON and STOP on d 1, and greater (P=0.04) in TRANS compared to CON on d 4. Serum NEFA concentrations were greater (P<0.01) in TRANS compared to CON and STOP on d 1, and greater (P≤0.05) in TRANS compared to CON on d 4 and 7. Mean plasma ceruloplasmin concentrations were similar (P=0.19) among treatments. Plasma haptoglobin concentrations were greater (P≤0.04) in TRANS compared to CON and STOP on d 1, and in STOP compared to CON on d 1. In conclusion, inclusion of rest stops during a 1,290-km transport prevented the increase in circulating cortisol and alleviated the NEFA and haptoglobin response elicited by transport, but did not improve feedlot receiving performance of transported cattle.

An assay for pro-oxidant reactivity based on phenoxyl radicals generated by laccase

A transient species may be detected with UV–vis and EPR spectroscopy during turnover of a laccase with quercetin; this species is assigned as a quercetin-derived radical, based on EPR spectra as well the observed UV–vis similarities (a 540nm centred band) with previously reported data. The rates of formation and decay of this species correlate well (r=0.9946) with the pro-oxidant reactivity manifested by flavonoids in the presence of laccase. An assay for the pro-oxidant reactivity of natural products is hence proposed based on the results reported here; its application is demonstrated for a series of pure compounds as well as for several propolis extracts. This assay has the advantages of using a biologically relevant process (haemoglobin oxidation), and not requiring the addition of oxidising agents such as peroxide or superoxide. Correlations, or the lack thereof, between the pro-oxidant parameters and the redox potentials, antioxidant capacities and lipophilicities, were analysed. The laccase employed in our study does display reactivity-related similarities to a range of other proteins, including human plasma ceruloplasmin.

Effects of trace element supplementation on apparent nutrient digestibility and utilisation in grass-fed zebu (Bos indicus) cattle

Trace element deficiencies in cattle are omnipresent, both in developing and industrialised regions. Little information is available on the effect of dietary trace elements on nutrient digestibility and utilisation, in spite of many deficiency-related symptoms suggesting a relevant role, such as loss of appetite in Zn deficiency and severe diarrhoea in Cu deficiency. The present study aimed to identify the early effects of dietary trace elements on nutrient utilisation in grass-fed zebu (Bos indicus) cattle. Adult bulls (n=8) were randomly assigned to a treatment: control or trace element supplementation (Zn, Mn, Cu, Se, I and Co) during 28 days. Grass mineral analysis suggested deficient Cu (5.53–9.60mg/kg) and Se (0.02–0.09mg/kg) concentrations in combination with high S (2577–3855mg/kg) and Mo (1.52–3.12mg/kg) and very high Fe (619–1214mg/kg) concentrations. Supplementation increased plasma Cu (0.82 vs. 0.61mg/l), Zn (1.40 vs. 0.89mg/l), Mn (0.30 vs. 0.05mg/l) and Se (0.07 vs. 0.06mg/l) concentrations (all P<0.05). Faecal Cu, Zn, Mn and Se were also increased (P<0.05), as was faecal Co (P=0.05) concentration in supplemented bulls. On the contrary, trace element supplementation did not affect plasma ceruloplasmin and superoxide dismutase activities (P>0.05). Also, no effects on apparent nutrient (dry matter, ash, protein, fat, and fibre) digestibility, apparent trace element absorption (except for Se and I) or plasma acyl carnitines (indicators of available energy substrates) were observed in this study (all P>0.05). Overall, despite clear improvement in trace element status – notwithstanding high concentrations of Cu antagonists in the grass diet – supplementation did not affect nutrient digestibility or utilisation in grass-fed zebu cattle.