Assessment of the Zinc and Copper Status in Alpaca.

This study was performed with the aim of investigating the concentration of zinc and copper in the blood of healthy alpacas (Vicugna pacos) kept in central Europe and to compare the concentration of Zn and Cu in plasma and in whole blood. A further objective was to evaluate blood Zn and Cu in relation to different micromineral supplementation, age and sex groups of alpacas. A total of 299 alpacas (224 adults and 75 crias) from 18 farms were included in this study. The concentrations of copper and zinc in plasma/whole blood were measured by flame atomic absorption spectrometry. The results of this study show high individual variability in plasma Zn (median 3.54, range 1.56-8.01 µmol/l), whole blood Zn (median 10.01, range 6.23-75.0 µmol/l), plasma Cu (median 7.53, range 2.93-16.41 µmol/l) and whole blood Cu (median 6.33, range 3.02-13.95 µmol/l). Plasma Zn was not significantly influenced by sex, age or feeding group. Whole blood Zn was only significantly higher in females than in males. The intake of Zn in all groups was equal to or higher than the nutritional recommendation. During excessive supplementation, Zn absorption decreased and thus blood Zn did not reflect the higher intake. Only a weak correlation was found (Spearman correlation coefficient r = 0.384; p > 0.01; n = 204) between plasma and whole blood Zn concentrations. Plasma copper concentration was significantly influenced by age, sex and feeding; whole blood Cu by age and feeding. However, neither plasma Cu nor whole blood Cu reflected the intake of the element. We found a close correlation between plasma and blood copper concentrations (Spearman correlation coefficient r = 0.9043; p ≤ 0.01; n = 99). According to our results, copper in plasma or blood is not a good indicator of copper intake.

Impact of acute metabolic acidosis on the acid-base balance in follicular fluid and blood in dairy cattle.

Acid-base balance is one of the most vigorously regulated variables of the body, including genital organs. Subacute ruminal acidosis is a common disturbance in dairy cows that disturbs several biochemical indices in the blood, cerebrospinal fluid, and urine. The possible negative effects of metabolic acidosis on the follicular fluid (FF) composition and, subsequently, on oocyte quality, are not fully elucidated. This study aimed to evaluate the changes in acid-base balance (ABB) in FF and blood during acute metabolic acidosis in dairy heifers. Ten Holstein heifers were stimulated with FSH in eight decreasing doses at 12-hour intervals (D0-D3). Acidosis was induced by oral administration of sucrose at 9 g/kg of body weight, dissolved in 10 L of warm tap water, at D3. Samples were collected from each cow at 0, 8, 12, 16, 24, 32, 40, and 48 hours after treatment. Samples of FF, obtained by transvaginal follicular aspiration, and peripheral blood were examined for ABB parameters: pH, pCO2, pO2, HCO3-, and base excess (BE). A significant decrease in pH, HCO3-, and BE values in the blood, as well as FF, occurred after sucrose treatment. The lowest pH values occurred in blood at 16 hours, and in FF at 24 hours, after treatment (7.30 ± 0.05 and 7.33 ± 0.05, respectively). The lowest HCO3- values in blood (18.75 ± 3.2 mmol/L) and FF (18.07 ± 2.84 mmol/L) occurred 24 hours after treatment, as did the lowest BE values (-6.61 ± 3.7 mmol/L and -7.53 ± 3.89 mmol/L, in blood and FF, respectively). Significant correlations for HCO3- (r = 0.928), BE (r = 0.946), pH (r = 0.889), and pCO2 (r = 0.522) existed between blood and FF samples. The results demonstrated that metabolic acute acidosis substantially influences the characteristics of both serum and FF.

Biological Effects of Drug-Free Alginate Beads Cross-Linked by Copper Ions Prepared Using External Ionotropic Gelation.

External ionotropic gelation offers a unique possibility to entrap multivalent ions in a polymer structure. The aim of this experimental study was to prepare new drug-free sodium alginate (ALG) particles cross-linked by Cu2+ ions and to investigate their technological parameters (particle size, sphericity, surface topology, swelling capacity, copper content, release of Cu2+ ions, mucoadhesivity) and biological activity (cytotoxicity and efficiency against the most common vaginal pathogens-Herpes simplex, Escherichia coli, Candida albicans) with respect to potential vaginal administration. Beads prepared from NaALG dispersions (3 or 4%) were cross-linked by Cu2+ ions (0.5 or 1.0 M CuCl2) using external ionotropic gelation. Prepared mucoadhesive beads with particle size over 1000 µm exhibited sufficient sphericity (all ˃0.89) and copper content (214.8-249.07 g/kg), which increased with concentration of polymer and hardening solution. Dissolution behaviour was characterized by extended burst effect, followed by 2 h of copper release. The efficiency of all samples against the most common vaginal pathogens was observed at cytotoxic Cu2+ concentrations. Anti-HSV activity was demonstrated at a Cu2+ concentration of 546 mg/L. Antibacterial activity of beads (expressed as minimum inhibition concentration, MIC) was influenced mainly by the rate of Cu2+ release which was controlled by the extent of swelling capacity. Lower MIC values were found for E. coli in comparison with C. albicans. Sample ALG-3_1.0 exhibited the fastest copper release and was proved to be the most effective against both bacteria. This could be a result of its lower polymer concentration in combination with smaller particle size and thus larger surface area.

Reference values for biochemical parameters in blood serum of young and adult alpacas (Vicugna pacos).

The aim of this study was to establish reference interval for biochemical parameters in blood of alpacas on the basis of large population of clinically healthy animals, and to determine the influence of sex, age and season on nitrogen and lipid metabolites, enzymes, electrolytes, vitamins and minerals in blood of alpacas. Blood samples were collected from 311 alpacas (61 males and 201 females >6 months of age and 49 crias (21 males and 28 females) ⩽6 months of age). Selected farms were located in Central Europe (Czech Republic and Germany). We determined 24 biochemical parameters from blood serum. We performed the comparison of results by the sex of animals and for the older group also the comparison of the results with regard to the season, respectively, to the feeding period. We found no highly significant difference (P<0.01) between males and females with the exception of γ-glutamyl transferase (GGT), alkaline phosphatase (ALP) and cholesterol. We found 15 significantly different parameters between the group of crias 6 months of age and the older alpacas. Based on our findings we suggest for most parameters to use different reference intervals (especially ALP, cholesterol, total protein, globulin, non-esterified fatty acids (NEFA), GGT and phosphorus) for the two above-mentioned age groups. Another important finding is the differences between some parameters in older group of alpacas in summer/winter feeding period. Animals in the summer feeding period have higher values of parameters related to fat mobilization (ß-hydroxybutyrate, NEFA) and liver metabolism (bilirubin, alanine aminotransferase). The winter period with increased feeding of supplements with higher amount of fat, vitamins and minerals is characteristic by increased values of cholesterol, triglycerides, vitamins A and E, and some minerals (K, Ca, Mg and Cl) in blood serum. Clinical laboratory diagnosis of metabolic disturbances may be improved with use of age-based reference values and with consideration of seasonal differences.

The influence of supplementation of different forms of zinc in goats on the zinc concentration in blood plasma and milk.

The aim of the experiment was to study zinc concentration in goat's milk during a short-term oral supplementation of three organic and one inorganic form of zinc and the influence of Zn supplementation on the concentration of Zn and Cu in blood plasma. The experiment involved 31 lactating goats. The group C was the control, group L received zinc lactate, group A received zinc chelate, group T received amino acid-polypeptidic complex with zinc, and group O received zinc oxide. Zinc preparations were administered orally in 500 mg Zn per animal for 28 days. In the course of the experiment, Zn concentration in plasma increased significantly in all experimental groups and in the control group was unchanged. The concentration of zinc in milk during the experiment fluctuated in individual groups, but there were no significant differences between groups. After 28 days of Zn supplementation, Zn concentration in milk of goats was similar as at the start of the experiment in all groups. According to our results, the supplementation of different forms of zinc did not influence the concentration of Zn in milk, but significantly influenced the Zn concentration in blood plasma. The efficiency of different organic and inorganic forms of zinc was similar.

[Diagnosis of magnesium deficiency in the body, personal experience]. / Diagnostika deficitu horcíku v organismu, vlastní zkusenosti.

Magnesium as a component of a range of enzymatic systems is a very important intracellular cation in the organism. Its monitoring is limited in many observations only to determination of its concentration in blood serum. We have done an analysis of serum and erythrocyte magnesium concentrations in 23 healthy women, 70 healthy men and in 184 patients with ulcerative disease of gastroduodenum. It was proved in all the monitored groups that erythrocyte concentrations of magnesium were lower compared to values which were estimated from serum concentrations in 9 from 23 healthy women, in 3 from 60 healthy men, in 10 from 101 ill men with ulcerative gastroduodenal disease, and in 17 from 83 women with ulcerative gastroduodenal disease. The highest rate of low concentrations of erythrocyte magnesium in healthy women with physiologic concentrations of magnesium in serum was also confirmed by currently created subgroup of healthy women (n = 11) who undergone together with other analysis peroral Mg2+ load test. This test confirmed magnesium deficiency in 10 from 11 women. The results showed there are more frequent deficiencies of magnesium in organisms then it is generally assumed. They also proved the importance of nutrition and regular food in population of healthy, young women.

Chromium supplementation enhances antibody response to vaccination with tetanus toxoid in cattle.

Nineteen multiparous late-pregnant dairy cows were divided into an experimental group (n = 10) and a control group (n = 9). Animals in the experimental group were fed a diet supplemented with chromium chelate admixed into wheat meal at 5 mg per animal per day. The supplementation was started 4 weeks before calving and stopped by the end of week 3 after calving. All the cows were vaccinated with tetanus toxoid in the fifth and sixth week of lactation. Monitoring of the characteristics of non-specific immunity did not show any significant between-group differences in total and differential leucocyte counts, percentages of lymphocyte subpopulations, activities of lectin-stimulated lymphocytes, phagocytic activities, and the contents of total immunoglobulins and lysozyme in blood sera. Tetanus toxoid-specific antibody titres, those of the IgG2 isotype in particular, were higher in the experimental group than in the control animals. These results indicate that chromium supplementation at the level used in this experiment modulated the regulation of functions of the immune system.

Histochemical and ultrahistochemical localization of heavy metals in calf organs.

Cadmium, zinc, selenium, and copper were administered, singly or in combination, orally or subcutaneously. Experiment I included 32 calves of both sexes; six received Cd (two groups), Zn, Cd, and Zn, and Cd and Se (two groups) and one group was a control. In Experiment II (21 bulls), three were given Cd, Cd, and Cu, and Cd and Zn, respectively, and one group was a control. For light microscopy, in Experiment I the highest amounts of silver granules were present in the samples of liver, small intestine, and vesicular gland of all the exposed groups; in Experiment II the most affected organs were liver, kidney, and small intestine. For electron microscopy, in Experiment I, after administration of Cd and Zn, the highest amounts of granules were seen in the cytoplasm of hepatocytes and cells of the proximal and distal renal tubules and the lowest amounts were found in glandular cells of the pancreas. Administration of Cd and Se resulted in the presence of large numbers of granules in the nuclei and nucleoli of spermatogonies. In Experiment II, ingestion of Cd and Zn in feed led to the appearance of highest amounts of granules in the nucleoli, nuclei, and cytoplasm of cells in testes, kidneys, and pancreas. Following Cd intake, the highest accumulation of granules was observed in the nucleoli of hepatocytes and cells of the proximal and distal renal tubules. Combined Cd and Cu produced the highest number of granules in cells of the proximal and distal renal tubules and in the nucleoli and nuclei of germinal epithelium.

Distribution of exogenous heavy metals in the hepatocytes of calves: a morphometric study.

The objective of this study was to evaluate the presence of heavy metals in the hepatocytes of the animals fed a cadmium-supplemented diet and also receiving zinc and/or selenium in the injection form. The experiment involved four groups of calves (6-8, both sexes) receiving the heavy metals in various combinations for 95 days. Electron micrographs of liver cells were prepared and statistically evaluated using Student's t-test. A modified morphometric apparatus was used for morphometric examination. Exogenous cadmium showed marked accumulation in the hepatocytes. If, however, the cadmium diet was combined with zinc or selenium administration the amount of the reduction product was much lower.

Towards the industrial production of medicinal tincture by ultrasound assisted extraction.

Experiments related to the industrial production of medicinal tinctures of sage (Salvia officinalis L.) and valerian (Valeriana officinalis L.) were performed using ultrasonically assisted extraction and quality of the final tincture was evaluated by gas chromatography and dry residue determination. The constituents of the active sage compound: borneol, cineole, alpha- and beta-thujone and also the dry residue were measured. In the case of valerian extractions, the efficiency of sonicated extraction was compared with classical extraction. The course of only the dry residue was monitored. The influence of ultrasound on the quality of valerian tincture was examined by HPLC.

[Determination of pentoxifylline and its major metabolites in human plasma using HPLC]. / Stanovenie pentoxifylínu a jeho hlavného metabolitu v plazme l'udí s vyuzitím HPLC.

The present paper describes a rapid, reliable and simple method of pentoxiphyllin in plasma. The worked-out one-step extraction with dichloromethane yields 63% recovery of pentoxiphyllin, 58% recovery of the metabolite and 54% recovery of the internal standard. The employed column with a reverse load of Spherisorb ODS, mobile phase 65% of 0.01 M KH2PO4 and 35% acetonitrile, detection at the wavelength lambda = 273 nm, makes it possible to achieve determination of pentoxiphyllin at the levels commencing with 5 ng.ml-1 of plasma. Reproducibility of determination is 6.8%, or 4.5% in the pentoxiphyllin concentrations of 50, or 100 ng.ml-1.

Optimization of fluorescence detection for the determination of labetalol in plasma by high-performance liquid chromatography.

A sensitive reversed-phase high-performance liquid chromatographic method for the determination of labetalol has been developed. A mobile phase consisting of citrate buffer (pH 6.5), acetonitrile and 2-propanol and an RP-8 column were used. The sensitivity of the fluorescence detection was enhanced to 1 ng/ml of labetalol in plasma by optimizing the emitted light. General guidelines for optimization of fluorescence detection are discussed.