Comparison of two automated spectrophotometric methods for ceruloplasmin measurement in pigs

Ceruloplasmin (Cp) determination could provide an objective measure of the health status of an animal and could be used as marker of animal health and welfare (Skinner, 2001) (Skinner, J. G., 2001. Special report. International standardization of acute phase proteins. Veterinary Clinical Pathology 30, 2-7.) but only manual methods have been reported to determine Cp concentrations in pigs (McCosker, 1961; Toussaint et al., 1995; Eckersall et al., 1996) (McCosker, P. J., 1961. Paraphenylenediamine oxidase activity and copper-levels in mammalian plasmas. Nature. 190, 887-889; Toussaint, M. J. M., Van Ederen, A. M., Gruys, E., 1995. Implication of clinical pathology in assessment of animal health and in animal production and meat inspection. Comparative Haematology International 5, 149-157; Eckersall, P. D., Saini, P. K., McComb, C., 1996. The acute phase response of acid soluble glycoprotein, alpha(1)-acid glycoprotein, ceruloplasmin, haptoglobin and C-reactive protein, in the pig. Veterinary Immunology and Immunopathology 51, 377-385). In the present study two automated methods based on the use of two different substrates for the determination of serum ceruloplasmin in pigs were developed, evaluated and compared. Both methods showed a good precision and detection limits, with no signs of inaccuracy and could be applied to biochemical autoanalyzers usually found in clinical laboratories using only minimal amounts of serum. Additionally the behaviour of Cp in experimental and clinical situations was studied showing an increase of around two fold after turpentine administration and significantly higher values in cases of pigs with inflammatory conditions when compared with healthy pigs.     

Effect of different level and source of copper supplementation on immune response and copper dependent enzyme activity in lambs

An experiment was conducted on 30 male Nellore lambs with average body weight (BW) of 15.45 +/- 0.06 kg to determine the level of the copper (Cu) supplementation in diet from inorganic and organic sources required for optimum immunity and its effect on copper dependent enzymes by allotting them randomly to five groups in completely randomized design. The dietary treatments were viz., basal diet (no Cu supplementation, BD), other four groups were offered BD supplemented with 7 or 14 ppm Cu from copper sulphate (CuSO(4)) and Cu-proteinate, respectively. The lambs were fed the respective diets at 3.5 per cent BW to meet the requirements except Cu for 180 days. The humoral immune response against Brucella abortus and chicken RBC was assessed after 90 days of feeding. The in vivo delayed type hyper sensitivity reaction against PHA-P and in vitro lymphocyte proliferation against Con A indicative of cell mediated immune response (CMI) was assayed at 180 days of feeding. At the end of experiment four lambs from each group were slaughtered for estimation of liver superoxide dismutase activity (SOD). The ceruloplasmin and RBC-SOD activities were higher (P < 0.05) in 14 ppm Cu supplemented lambs from Cu-proteinate at 90 and 180 days, while the liver SOD activity was higher (P < 0.05) in lambs fed 14 ppm Cu from CuSO(4). The STAT titres against B. abortus were higher in Cu supplemented lambs, with no effect of dose of supplementation. Lambs supplemented with Cu-proteinate had higher titers than CuSO(4) on 7 and 14 days of post sensitization. The total immunoglobulin concentration and the CMI response against PHA-P and Con-A was higher (P < 0.05) in lambs fed 14 ppm Cu-proteinae diet. The IgM level was though high in Cu supplemented lambs, no dose or source effect were observed. The study indicated that Cu dependent enzymes activity and immune response were highest and respond better against stress in lambs on 14 ppm supplemented Cu from Cu-proteinate.     

日粮中不同钼铜水平对肉牛血液生化指标的影响

本研究旨在探讨日粮中不同钼铜水平对肉牛血液生化指标的影响。选用性别一致、年龄、体重相近(340±10)kg的3头健康未去势公牛作为试验动物。试验采用3×3拉丁方设计,分2期进行,第一期日粮钼的水平为0.27 mg/kg DM,第二期钼的水平为5.27 mg/kg DM。每期日粮铜的添加量分别为:10 mg/kg(低铜组),25 mg/kg(中铜组),50 mg/kg(高铜组)3个水平。颈静脉采血测定血铜、血清铜蓝蛋白(ceruloplasmin,Cp)和血浆中过氧化物歧化酶(superoxide dismutase,SOD)的活性。试验结果表明:在同一钼水平下,随铜水平的增加,牛体内血铜含量、血浆中SOD活性和血清Cp的活性逐渐增加。其中钼水平为0.27 mg/kg时,这种变化不显著(P>0.05)。钼水平为5.27 mg/kg时,添加10 mg/kg铜,血铜含量和血浆中SOD活性显著小于其他2个铜水平(P<0.05),Cp活性小于其他2个铜水平,且差异极显著(P<0.01)。在同一铜水平下,低钼组2种酶的活性普遍高于高钼组。日粮钼水平为0.27 mg/kg时,添加10 mg/kg铜可满足机体需要;日粮钼水平为5.27 mg/kg时,添加25 mg/kg铜较适宜:牛在缺铜时,与血浆SOD的活性比较,血清Cp更宜作为评价铜营养缺乏的指标;铜满足机体需要后,血铜、血清Cp和血浆SOD均对铜和钼的变化不敏感。血铜和血清铜蓝蛋白与日粮铜水平呈强相关。     

Copper status and enzyme, hormone, vitamin and immune function in heifers

A survey was conducted in 10 districts of Northern India to record the copper deficiency in soil, fodder and serum samples. Significant deficiency of copper was observed in soil, fodder and serum samples of heifers. The copper deficient animals were listless, showed depigmentation of skin, stiff gait and were anaemic and diahorric. The purpose of the study was to evaluate the magnitude of copper deficiency in Northern India and to examine the various haematobiochemicals, enzymes, vitamin and immune function which are affected by the deficiency of copper, so as to identify the parameters which can be of diagnostic importance in copper deficiency. Forty hypocuperemic heifers were selected from these areas and were randomly divided into two groups A and B. The heifers in group A were provided with mineral mixture containing copper sulphate and in group B without copper sulphate. Significant improvement (P<0.01) was observed in the haemoglobin (Hb), total leukocyte count (TLC) and total erythrocyte count (TEC) level at the 30th day of treatment in the animals of group A. Significant (P<0.01) improvement in serum. Ceruloplasmin (Cp) level was observed within 30 days of treatment, while significant (P<0.01) improvement in monoamine oxidase and liver cytochrome oxidase was observed at the 60th day of treatment in group A animals. Regarding hormones significant improvement was observed in T(3) and T(4), in the animals of group A within 60 days of treatment. The values of vitamin A and E showed significant (P<0.01) improvement within 30th days of treatment. The phagocytic activity of neutrophils against Candida albicans significantly (P<0.01) improved in group A within 60 days of treatment. Similarly significant improvement in superoxide dismutase activity in RBC was observed at the 30th day, and WBC and whole blood at 60th day in group A animals. Significant improvement in liver Cu level was observed at the 30th day of treatment, while in group B the liver Cu was significantly (P<0.01) depleted at the 60th day of experimentation. Additional Cu supplementation improved growth performance significantly in group A.     

Hepatic (64)Cu excretion in Dobermanns with subclinical hepatitis

To investigate whether Dobermanns have impaired copper excretion an intravenous radioactive copper isotope ((64)Cu) was used as a tracer. Five patients and eight normal dogs (5 normal Dobermanns and 3 Beagles) were studied. The five female Dobermann patients had a subclinical hepatitis and an increased hepatic copper concentration (median 822mg/kg, range 690-1380mg/kg dry matter). The normal dogs, five Dobermanns and three Beagles, had no abnormal liver histopathology and hepatic copper concentrations were considered normal (Dobermanns; median 118mg/kg, range 50-242mg/kg dry matter; Beagles; median 82mg/kg, range 50-88mg/kg dry matter). Cholestasis was excluded in all dogs by means of a (99m)Tc-Bis-IDA hepatobiliary scintigraphy. Plasma clearance of (64)Cu was comparable in all dogs with no statistically significant differences. The excretion of (64)Cu into the bile, although not statistically significant, was less for the Dobermanns with subclinical hepatitis compared to the normal dogs. The findings suggest that impaired copper excretion may play a role in the aetiology of chronic hepatitis in the Dobermann.     

Antioxidant enzymes and somatic cell count in dairy cows fed with organic source of zinc,copper and selenium

The objectives of this study were to evaluate the effects of dietary organic sources of zinc (Zn), copper (Cu) and selenium (Se) for dairy cows on somatic cell count (SCC), occurrence of subclinical mastitis, number of clinical mastitis cases and concentration of superoxide dismutase (CuZnSOD), glutathione peroxidase (GSH-Px) and ceruloplasmin (CP). Nineteen dairy cows, with confirmed pregnancy, were selected by body weight, body condition score (BCS), number of lactation, and milk yield in previous lactation, and randomly distributed among two groups, one to receive organic (n = 9) and the other the inorganic (n = 10) source of Zn, Cu and Se. Diets were formulated to meet the nutritional requirements of animals from 60 days before the expected date of calving up to 80 days of lactation. Blood samples were collected at − 60, − 21, 1, 21, 40 and 80 days in relation to the expected date of calving for analysis of CuZnSOD, GSH-Px and CP concentration. Milk samples for SCC determination were collected weekly after 15 days of lactation, also at day 1 and 7 and when a clinical case was diagnosed for microbiological culture. The number of new and total cases of subclinical mastitis was lower for the group of cows fed with organic sources of Zn, Cu and Se compared to animals receiving inorganic sources. Average SCC during the first 80 days of lactation tended to be lower for the group fed with organic Zn, Cu and Se. Feeding organic Zn, Cu and Se did not alter the concentration of CuZnSOD, GSH-Px and CP.     

Selected Biochemical Indicators of Equine Rhabdomyolysis in Arabian Horses: Acute Phase Proteins and Trace Elements

Although creatine kinase (CK), aspartate transaminase (AST), cytokines, and oxidative stress parameters were shown to be useful biomarkers for diagnosis of equine rhabdomyolysis, additional biomarkers of the disease may be of interest to indicate prognosis of the disease. Therefore, the present study investigated acute phase proteins and trace elements as additional biomarkers of ER. Sixty male horses (4-6 years old) of 2 equal groups were used. Horses of the first group were clinically healthy and served as a control group, whereas horses of the second group were ER-diseased animals. Harvested sera were used for estimation of the activities of CK, AST, lactate dehydrogenase (LDH), serum amyloid A (SAA), haptoglobin (Hp), ceruloplasmin (Cp), copper, iron, and zinc, whereas plasma samples were used for determination of fibrinogen. The present findings revealed a significant (P ≤ .05) increase in values of CK, AST, and LDH in diseased horses compared to those in control values. In addition, a significant (P ≤ .05) increase in SAA (162.6 ± 5.32 mg/L), Hp (3.6 ± 0.54 g/L), Cp (39.32 ± 2.31 mg/L), and copper (28.36 ± 1.23 μmol/L) along with a significant (P ≤ .05) reduction in levels of iron (9.32 ± 0.23 μmol/L) and zinc (8.65 ± 1.02 μmol/L) was recorded in diseased horses compare to that in controls (11.3 ± 2.2 mg/L, 0.8 ± 0.2 g/L, 24.23 ± 1.32 mg/L, 18.41 ± 1.03 μmol/L, and 14.2 ± 0.42 μmol/L, respectively). In conclusion, SAA, Hp, Cp, copper, and zinc were useful prognostic biomarkers for the diagnosis of ER in Arabian horses.